American medical journal

The Impact of Gut Microbiota on Long COVID: Insights and Challenges

Introduction

The ongoing global COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in a spectrum of clinical manifestations, ranging from asymptomatic or mild respiratory symptoms to severe pneumonia and multi-organ dysfunction1. While the acute phase of the disease has been extensively studied, the emergence of a condition known as “Long COVID” or “Post-Acute Sequelae of COVID-19 (PASC)” has raised significant concern due to its persistent and debilitating nature. Long COVID is characterized by symptoms that persist for weeks or even months after the acute infection has resolved, affecting various organ systems and significantly impairing patients’ quality of life [1-3]. Recent research has shed light on the potential role of gut microbiota in the pathogenesis of Long COVID, as alterations in the composition and dynamics of the gut microbiome have been observed in patients with both acute COVID-19 and Long COVID [2,3]. The gut microbiota regulates the host’s immune response, metabolic processes, and overall health. Dysbiosis, or an imbalance in the gut microbiome, can lead to immune dysfunction and chronic inflammation, hallmarks of Long COVID [4].

This emerging connection between gut microbiota and Long-term COVID has prompted investigations into its potential as a therapeutic target for alleviating symptoms and improving outcomes in affected individuals [5-7]. Several studies have explored the dynamics of gut microbiota in patients with long-term COVID-19, shedding light on how changes in microbial composition may correlate with symptom severity and persistence2,3,5. Additionally, research has examined the impact of factors such as antibiotics, probiotics, and dietary interventions on gut microbiota in Long-term COVID patients. Understanding the relationship between the gut microbiome and Long-term COVID is crucial, as it may open new avenues for therapeutic interventions and improve the management of this complex and poorly understood condition [8,9]. In this comprehensive review, we aim to synthesize the existing literature on the role of gut microbiota in Long COVID. We will analyze findings from recent studies investigating alterations in gut microbiota composition, diversity, and function in patients with Long-term COVID and their potential implications for disease pathogenesis and symptomatology [9-11]. Additionally, we will explore the impact of various interventions, including dietary modifications and probiotics, on modulating the gut microbiome in Long COVID patients. Our objective is to provide a thorough overview of the current knowledge regarding the gut microbiota’s involvement in Long-term COVID and its potential as a target for therapeutic interventions.

Methods

The research methodology involved a comprehensive search of multiple reputable databases to ensure the inclusion of relevant studies while minimizing the risk of bias. PubMed, Scopus, Scielo, Embase, and Web of Science were chosen due to their comprehensive coverage of peer-reviewed literature in the medical field. Additionally, Google Scholar was utilized to access gray literature, which often includes valuable insights not found in traditional peer-reviewed articles. The study’s selection criteria were centered on the study’s focus, which was artificial intelligence’s impact on general surgeons’ training. To refine the search and capture relevant studies, a combination of keywords was used, including “post-acute COVID-19 syndrome”, “post-acute COVID-19 syndromes”, “long COVID, PASC post-acute sequelae of COVID-19”, “gastrointestinal microbiome,” and “gut microbiota.” This approach ensured that the selected studies were directly related to the topic of interest. The inclusion criteria encompassed various studies, such as systematic reviews, case-control studies, cross-sectional studies, case series, and review articles.

This broad inclusion criteria aimed to gather a comprehensive range of evidence and perspectives on the subject matter. The process of analysis, review, and selection of materials was conducted rigorously to maintain the quality and relevance of the chosen studies. It involved a systematic and blind approach, with pairs of reviewers independently assessing the title and abstract of each study. In cases of disagreement between the two reviewers, a third reviewer was involved to reach a consensus and ensure the final selection of studies was based on well-founded criteria. This meticulous research methodology guarantees that the findings and conclusions drawn in the article are rooted in a robust and diverse body of evidence, enhancing the credibility and reliability of the study’s outcomes.

Results and Discussion

Long COVID has emerged as a perplexing and debilitating condition characterized by persistent symptoms following acute COVID-19 infection. Recent research has shed light on the potential role of the gut microbiota in influencing the course of long-term COVID, offering intriguing insights into the consequences and effects of microbial alterations [12]. A recurring theme in these studies is the presence of dysbiosis within the gut microbiota of Long COVID patients. This dysbiosis typically involves a decrease in beneficial commensal bacteria like Bifidobacterium and Lactobacillus, coupled with an increase in pro-inflammatory taxa5-8. Such microbial imbalances are thought to trigger immune dysregulation and chronic inflammation, potentially contributing to the prolonged and diverse symptomatology observed in Long COVID4. One significant finding is the association between gut microbiota profiles and the severity and duration of Long COVID symptoms5. Liu et al. revealed that individuals experiencing more severe Long COVID symptoms exhibited distinct microbiota profiles, implying a potential link between gut microbiome composition and symptom severity [5,6].

This observation was further corroborated by Ancona et al., who reported dysbiosis in both gut and airway microbiota, suggesting its potential role in influencing respiratory symptoms associated with long-term COVID3. Recognizing gut microbiota involvement in Long COVID has opened doors to therapeutic exploration. Su et al. (2022) delved into the impact of antibiotics and probiotics on gut antimicrobial resistance gene reservoirs, highlighting the potential of these interventions to modulate gut microbiome composition and, consequently, Long-term COVID outcomes [7,11,12]. Furthermore, Wang et al. demonstrated the alleviation of severe gastrointestinal symptoms in a Long COVID patient through nutritional modulation of the gut microbiota, suggesting the promise of dietary interventions [13]. Intriguingly, parallels have been drawn between long-term COVID and myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS), prompting investigations into potential shared mechanisms4-6. Komaroff and Lipkin proposed that insights from ME/CFS research might contribute to unraveling the pathogenesis of long-term COVID-19, hinting at possible commonalities that potentially involve the gut microbiota [2-4]. Vestad et al. provided valuable insights into the persistence of gut microbiota alterations beyond the acute phase of COVID-19 [14-17]. Their study suggested that the impact of gut dysbiosis might extend for months, emphasizing the necessity for long-term monitoring and interventions to address the consequences of these microbial changes18. Mundula et al. explored the implications of chronic low-grade inflammation in various diseases, including the potential modulation through gut microbiota interventions.

This perspective highlights the crucial role of gut microbiota in regulating host inflammatory responses and its potential influence on disease outcomes19. Moreover, Nguyen et al. conducted a metagenomic assessment of gut microbial communities, offering a deeper understanding of their potential influence on COVID-19 severity15. Their study unraveled the intricate interplay between gut microbiota and host responses, providing critical insights into how microbiota composition may shape the course of the disease [18-20]. While these studies collectively suggest a role for gut microbiota in long-term COVID, it is essential to acknowledge the variability in findings and methodologies. Robust and standardized research is necessary to establish causality and unravel the intricate mechanisms by which the gut microbiota influences long COVID.

Conclusion

In conclusion, the emerging evidence underscores the potential significance of gut microbiota alterations in the pathogenesis and clinical progression of long-term COVID-19. Dysbiosis in the gut microbiome appears to be associated with symptom severity and duration, offering promising avenues for therapeutic interventions.

Longitudinal studies indicate that the impact of gut dysbiosis may extend well beyond the acute phase of the disease. However, comprehensive, and standardized research is crucial to establish causality definitively and unlock the intricate mechanisms underlying the influence of the gut microbiota on COVID. Understanding this relationship holds the promise of novel approaches to managing and improving the outcomes of Long COVID patients, addressing a pressing public health concern.

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Journals on Biomedical Intervention

Synthesis and In Silico Analysis of Chalcone Derivatives as Potential Prostaglandin Synthetase Inhibitors

Summary

Prostaglandins (PGs) are biochemical endogenous lipids with autacoid functions, synthesized in- vivo from arachidonic acid [1]. PGs and other similar physiologically active compounds are collectively known as metabolites of eicosanoids [2]. PGs have long been reported to have sustained homeostatic roles and facilitate many pathogenic mechanisms in inflammation, gastrointestinal tract [1], muscle contraction, blood clotting [3], ocular protection [4,5], and regulation of the circulatory system [6,7]. These lipids are produced via the action of the cyclo-oxygenase (COX-1 and COX-2) isoenzymes, and their biosynthesis is antagonized by the nonsteroidal anti-inflammatory drugs (NSAIDs) [1]. Some vitamins, including D3 (cholecalciferol), and K2 (menaquinone) are also known to inhibit the actions and biosynthesis of PGs [8-10]. PGs mainly take part in vasodilation, conception, luteolysis, menstruation, parturition, blood pressure reduction, control of sodium reabsorption by the kidney, etc [11]. Studies have shown that excessive concentrations of PGs induce diarrhea that accompanies medullary carcinoma of the thyroid or neural crest tumors and mediates several inflammatory responses [11], incoordinate hyperactivity of the uterine muscle leading to uterine ischemia, and menstrual cramps in women [12]. PG structural analogs like latanoprost, travoprost, and bimatoprost with antagonistic properties, are being used and well-tolerated for the reduction of intraocular pressure (IOP) in patients with primary open-angle glaucoma and ocular hypertension [7].

Previous reports have indicated that there are some correlations between high levels of PGs analog (PGFS) in tumors of the GI tract and the effectiveness of NSAIDs [13]. Thus, they can be used in the study, design, and discovery of antitumor agents. Cyclooxygenase 1 and 2 (COX-1 and COX-2) biologically transform arachidonic acid (AA) to prostaglandins H2 (PGH2), which is further biotransformed to various PGs, and other endogenous lipids like thromboxanes, leukotrienes, and hydroxyeicosateraenoic acids [14,15]. The names of these enzymes are derived from their catalytic cyclo-oxygenation that converts AA to prostaglandin G2 (PGG2), and peroxidation of PGG2 to PGH2, hence are also known as peroxidase enzymes as well [16]. The three COX isoforms, COX-1 COX-2, and COX-3, have been identified to share almost 60% amino acid sequence similarity but with much higher sequence homology in the catalytic sites (Figure 1) [17]. COX-1 is firmly expressed in many tissues, while COX-2 is strongly induced by various mitogens and plays imperative roles in many pathological conditions like inflammation [18,19]. There is also a COX-3 enzyme but reported to be not functional in humans. COX-3 isozyme is encoded by the same gene as COX-1, but COX-3 retains a particular nucleotide sequence (intron 94) that is not retained in COX-1 [20].

PGE2 increases gastric mucus secretion, uterus contraction (particularly during pregnancy), GI tract smooth muscle contraction, inhibition of lipolysis, autonomic neurotransmitters regulation, platelet response to agonists, and in-vivo atherothrombosis [21,22]. COX-1 enzyme regulates the baseline levels of PGs, while COX-2 synthesizes PGs via stimulation and significantly increases PGs levels during growth and inflammation, although both enzymes are located in the stomach, kidneys, and blood vessels [23]. Hence, inhibition of these agents is necessary for the optimal regulation of many biological functions, when in excessive amounts. NSAIDs inhibit the activities of COX-1 and COX-2 enzymes. There are non-selective (inhibits both COX-1 and COX-2), and COX-2 selective NSAIDs. These NSAIDs, while reducing inflammation caused by PGs, also inhibit platelet aggregation and increase the risk of GIT ulcers and intestinal bleeding [24]. COX-2 selective inhibitors promote thrombosis and increase the risk of heart attacks [25]. Due to these adverse reactions, coupled with the high etiology of vascular, and kidney disease complications, some COX-2 inhibitors are no longer in clinical use [26]. There are also reports that NSAIDs impair the production of erythropoietin, resulting in anaemia [27].

The long-term harmful effects of most NSAIDs outweigh the medical benefits. A study conducted a few years ago, observed a statistically significant increase in myocardial infarction incidence among patients on rofecoxib [28], and data from approved clinical trials, showed a significant relative risk of cardiovascular events that led to the global withdrawal of rofecoxib in 2004 [29]. Another study reported a significant increase in erectile dysfunction in men who frequently used NSAIDs [30]. NSAIDs are also associated with a doubled risk of heart failure in people who have not experienced cardiac disease in their lifetime [31]. Finally, the use of NSAIDs during late pregnancy can cause miscarriage [32], premature birth [33], constriction, and closure of fetal ductus arteriosus, leading to different blood-related congenital heart diseases in the fetus [34]. Acetaminophen, regarded as the safest, and well tolerated NSAID during pregnancy, was reported to cause male infertility in the fetus [35,36]. These advents call for the search for more effective, with minimal toxic molecules that can be used clinically to alleviate inflammatory conditions.

Chalcones chemically known as 1,3-diaryl-2-propen-1-ones, are flavonoids and isoflavonoids precursors, are chemical moieties present in many naturally compounds and are also prepared synthetically because of their convenient synthetic procedures [37]. Chalcone derivatives have been reported to possess antiproliferative [38], anti- inflammatory [39], antitumor [40], antimalarial [41], antibacterial [42], antiviral [43], antileishmanial [44], antifungal [45] properties, among others [46]. Molecular docking is a veritable tool used in the computational prediction of ligands and protein inhibitory affinity in the search for lead molecules [47], including characterized natural products [48]. Therefore, we conducted the synthesis and molecular docking of some chalones derivatives which biological properties were evaluated previously, that can serve as lead compounds in the design of anti- inflammatory and analgesic agents, especially as potential prostaglandin synthetase enzymes (COX-1 and COX-2) secretagogues inhibitors.

Method

Synthesis

Scheme 1: Synthesis of Methoxy, Halogenated and Aminated Chalcone Derivatives: All reagents used in the synthesis and other analysis were of analytical grades. The IR data was obtained from the FTIR-8400S instrument, Shimadzu global links, North America, while Nuclear Magnetic Resonance (NMR) experiment was performed on a 400 MHz instrument, obtained from Varian Inc. Palo Alto, California, USA. An equivalent weight of 10.6 g benzaldehyde and 12.0 g acetophenone were weighed, into a 100 ml flask having 25 mL EtOH, and stirred on ice (4-0℃). 20 ml of KOH (20%) was added with continuous stirring for 20 minutes and allowed to stand for 24 hours. Ice chips were added, and the mixture was titrated with 25 mL of 20% acetic acid (4-0℃). Precipitates were formed, filtered under suction and recrystallized with ethanol, dried, the percentage yield and melting point were determined. This procedure was repeated with different benzaldehyde derivatives, including para-methoxybenzaldehyde, para-chlorobenzaldehyde and para-dimethylaminobenzaldehyde, giving rise to various derivatives of chalone (Scheme 1).

Scheme 2: Synthesis of 6-Diphenyl-2-Thiopyrimidine Chalcone Derivatives: From the initial 4-methoxy-chalone derivative obtained, an equivalent weight of 2.38 g 4- methoxy-chalone, 2.12 g sodium bicarbonate, and 1.52 g thiourea, were weighed into a flask having 30 mL DMSO. The mixture was refluxed under Nitrogen gas for about 2 hrs, using a Thin Layer Chromatographic plate to monitor the progress of the reaction. Water was added to the reaction medium at the end and was allowed to stand for 24 hours. Precipitates were formed, filtered under suction, the residues were recrystallized with diethyl-ether and petroleum spirit. The percentage yield and melting points of the crystals obtained were quantified, after drying (Scheme 2).

Scheme 3: Synthesis of Epoxide Chalcone Derivatives: For the synthesis of epoxide derivatives, an equivalent weight of 5.16 g chalcone obtained previously was weighed into a beaker; 10 ml of 10% NaOH and 60 ml of MeOH were added respectively. The content of the beaker was dissolved with stirring via gentle heat, then 20 ml hydrogen peroxide H2O2) was added and stirred for 30 minutes. 5 ml of 10% acetic acid was used to acidify the medium. The resultant product was collected, and recrystallized with MeOH, filtered and dried. The percentage yield and melting points were respectively determined. This procedure was repeated with para-methoxychalcone, para-chlorochalcone, para- dimethylaminochalone, respectively, leading to the production of chalcone epoxide derivatives (Scheme 3).

Molecular Docking

Molecular modeling and docking simulations of the binding protein and synthesized ligands were done using the Maestro software of OPLS3, 2018 Force field [49], and Pymol software [50]. The docking parameters and affinity were compared with the previously reported pharmacological profile of the chalcone derivatives. The human COX-1 crystal structure protein (6Y3C) [51], and 1PXX (COX- 2 crystal structure with diclofenac bound to the cyclooxygenase active site) [52], were obtained from the PDB website, and modeled with the Pymol and D3Pocket webserver [53,54], to obtain all possible binding pockets and utilize one(s) with the highest affinity using diclofenac and Celecoxib (a selective COX-2 inhibitor) as the standard molecules.

A. (E)-Chalcone.
B. Para-chlorochalcone [(E)-3-(4-chlorophenyl)-1-phenylprop- 2-en-1-one]
C. Para-methoxychalcone [(E)-3-(4-methoxyphenyl)-1-phenylprop- 2-en-1-one]
D. Para-dimethylaminochalcone [(E)-3-(4-(dimethylamino) phenyl)-1-phenylprop-2-en-1-one]
E. Para-methoxy-4,6-diphenyl-2-thiopyrimidine[4-(4-methoxyphenyl)- 6-phenyl-5,6 dihydropyrimidine 2(1H)-thione]
F. Chalcone-epoxide [phenyl(3-phenyloxiran-2-yl) methadone]
G. Para-chlorochalcone-epoxide [(3-(4-chlorophenyl) oxiran- 2-yl) (phenyl)methanone]
H. Para-methoxychalcone-epoxide [(3-(4-methoxyphenyl) oxiran- 2-yl) (phenyl)methanone]
I. Para-dimethylaminochalcone-epoxide[(3-(4-(dimethylamino) phenyl)oxiran-2-yl)(phenyl)methanone]

Discussion

The compounds were obtained in high yield after the synthetic processes (Schemes 1 & 2). The percentage yields of the compound ranged from 27.68 – 90.38%, with sample B having the highest yield while sample I gave lowest synthetic yield. Also, the spectroscopic (FTIR and NMR) analysis shows distinct spectrum across all molecules, indicating the presence of unique functional groups and chemical environments (Table 1). All the synthesized chalcones derivatives showed appreciable protein binding affinity against the COX-1 and COX-2 enzymes. Compared to the standard drugs, better protein-ligand affinity was observed with COX-2 enzyme. Diclofenac is known to inhibit both COX-1 and COX-2 enzymes [55]. The computational experiment showed that, some of the synthesized compounds had higher binding affinity against the COX-1 protein than both diclofenac and celecoxib. Compound A showed the highest affinity (-7.24 kcal/mol), while other compounds had affinity level of -7.21(C), -7.16 (B), -7.14 (I), -7.10 (D), -7.00 (H), -6.92 (G), -6.88 (F), and -6.11 kcal/mol(E), respectively. Whereas, the standard compounds (diclofenac and celecoxib) had -5.46 and -6.19 kcal/mol, respectively.

The protein-ligand interactions showed the actual protein residues in the COX-1 protein that compounds bound with (Figure 2). Also, the binding pocket and pose of the compound with highest affinity showed how it is well fitted into the protein pocket (Figure 3). Celecoxib, a selective COX-2 inhibitor showed highest binding affinity of -10.55 kcal/mol, while the test compounds had -8.55 (A), -8.84 (B) -8.50 (C), -8.79 (D), -8.24 (E) -8.13 (F), -8.50 (G), -8.22 (H), -8.69 (I), and diclofenac had -8.49 kcal/mol respectively (Table 2). The binding interactions of all molecules are shown in Figure 4, while binding poses of the molecules with the highest affinity is illustrated in Figure 5. Compounds E (4-methoxy-4,6-diphenyl-2-thiopyrimidine) and B (para-chlorochalcone) from previous studies, displayed remarkable anti-inflammatory in an in-vivo analysis using animal model [39]. E also showed the appreciable affinity against COX-1 protein more than the standard compounds, while lower affinity was observed against COX-2 protein. For compound B, it showed the highest affinity against COX-2 and very high affinity towards COX-1 protein compared to the standard molecules used in the analysis. This shows that with adequate physiochemical and structural modifications, these compounds could serve as potential lead compounds in analgesic and anti-inflammatory pharmacology, as pain and inflammation are associated with these enzymes in the biological system [16].

Conclusion

Analgesic and anti-inflammatory agents are used in the prevention of all kinds of pains, ranging from minor headaches to severe post-operative pains. The search for newer agents due to poor tolerability, adverse reactions and affordability of the existing ones is the focus of contemporary drug design and development. The compounds showed highly promising results in both in-vivo and computational molecular docking studies. Hence, the compounds reported in this study could be utilized and further modified structurally and physiochemically to achieve better analgesic and anti-inflammatory properties.

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Journals on Biomedical Imaging

Impact of Die Configuration on Physio-Chemical Properties, Anti-Nutritional Compounds, and Sensory Evaluation of Multi-Based Extruded Puffs

Introduction

Millets are highly valued by vegetarians, vegans, and individuals with coeliac disease for their exceptional nutritional benefits [1]. These plant-based foods are abundant in proteins and complex carbohydrates while being low in fat [2]. Additionally, they are a rich source of dietary fiber, essential minerals, and B-group vitamins [3,4]. However, Millet preparation can be a time-consuming process as they require prolonged soaking and cooking, which reduces their consumption by consumers [5]. To overcome this challenge, novel food products like – ready-to-eat Millet-based snacks could be developed. Such products could encourage more individuals to incorporate Millet into their diet [6]. The food processing technique of extrusion- cooking is highly versatile, multifunctional, and cost-effective. Raw materials are exposed to heat, pressure and shear forces during this process, leading to various biochemical reactions such as protein denaturation, starch gelatinization, fiber degradation, amylose-lipid complex formation through Maillard reaction [7]. Extrusion-cooking is commonly employed to create expanded snacks that are ready-toeat, come in different shapes and textures, and boast enhanced flavour and colour [8]. High concentrations of raffinose-family oligosaccharides (RFOs) in lentils cause stomach discomfort and reduce lentil quality for human consumption.

To develop strategies for lentil quality improvement, variability, heritability and effects of environmental conditions on the content and composition of soluble carbohydrates in lentil seeds have been investigated in detail [9]. Over the past few years, there has been significant research on creating extruded products using Millets [10]. In most cases, these Millets are blended with wheat [11]. Numerous studies have examined the impact of various extrusion-cooking factors, including feed moisture, screw speed, extrusion temperature, nutritional characteristics [12] physico-chemical, sensory and textural properties of prepared product. These studies have also included trials conducted directly at the industrial level [13,14]. However, there has been a lack of research on how the die configuration, which determines the final product’s shape and size, affects the extrusion of Millets. Food shape and size play a critical role in capturing the consumer’s attention [15]. These features strongly influence the implicit associations with consumers concerning nutritional value [16]. Moreover, the shape and size of food can significantly impact the sensory qualities and physico-chemical properties of the extruded product [17]. Therefore, the present study was designed to examine the impact of star shaped and circular die configurations, which have two different diameters of 35.9 and19.6 mm², respectively, on the anti-nutritional component, physico-chemical properties, and sensory features of extruded snacks made from Millet flour.

Materials and Method

For the development of extruded puff products, germinated pearl Millet (Pennisetum glaucum), finger Millet (Eleusine coracana), sorghum (Sorghum bicolour), foxtail Millet (Setaria italic) and rice (Oryza sativa) as base were used as raw materials for the present study work. These Millets were procured from a nearby local market in Meerut, Uttar Pradesh [18].

Optimization of Single Screw Extruder Operating Parameters for Different Millet Based Extruded Puffs Products

The operating parameters of the twin screw extruder mainly, the temperature, feed moisture content and the screw speed, were optimized for the various “best selected” four Millet fortified with rice based extruded puff products formulation. Three feed moisture content (20%, 25%, and 30% wt.b.), screw speeds (180, 270, and 360 rpm) and 160, 180°C and 270°C temperature were selected to produce extrudes. The extruded samples were then analyzed for physical analysis like expansion ratio, water holding capacity, moisture content, puffing properties and texture analysis. Further experiment was carried out at three different level of temperature (100, 110 and 120°C) with keeping constant screw speed and feed moisture content.

Sample Preparation from Extrusion

Laboratory scale single-screw extruder (Model no: GTL-100), (Zigmo Agro Pvt. Ltd. New Delhi, India) was used for sample preparation. During each extruder run, the extruder machine was allowed to equilibrate for 5-10 min until a stable torque was achieved. Extruded samples were collected on metal screens to allow excess steam to flash off. The extruded samples were collected in a low-density polyethylene bag after cooling and stored in a cool and dried area.

Development of Extruded Puff Products Based on the Millet’s Combinations

Extruded puffs preparation was done, and it mainly consists of puffs mixture of Millets in combination as shown in Tables 1 & 2 and rice is used as base. The blended puffs mixes at appropriate moisture content were extruded to produce extruded puff through the extrusion machine (manufacture: Jas Enterprises, Ahmadabad, India) into desired shaped products and developing extruded puff products based on Millet combinations involves a series of steps to create a desirable texture, flavor, and nutritional profile the process flow chart shown in Figure 1.

Multi Millet Conditioning

To achieve the optimal moisture concentration for extrusion (16 g/100 g), the Puffs were conditioned. The amount of water required to reach this moisture level was determined based on the initial moisture content of each flour type, which was 12.95 ± 0.01, 10.87 ± 0.01, and 12.54 ± 0.06 for pearl Millet, finger Millet and foxtail Millet, respectively. Water was added gradually to the flour in dough mixer at average speed to avoid the lumps formation. The process took approximately 20 minutes to obtain uniformly hydrated flour.

Extrusion-Cooking Process

Extrusion cooking is a food processing technology that integrates multiple operations such as mixing, cooking, kneading, shearing, shaping, and forming [19]. The DSE30 Lab Twin-screw extruder was used to carry out the extrusion-cooking process with a 12 kg/h capacity. The extruder machine design had two 38CrMoAl screws, a 5-kW motor and operating at maximum screw speed of 500 rpm, with three heating zones at 55, 95, and 125°C, respectively. The extrusion process was carried out using circular and star-shaped dies holes with cross-sections of 19.6 mm2 and 35.9 mm2, respectively. The circular die nozzle and star cross-section had a length of 6.35 mm. The feed rate was set to 2.5 g/s, the die temperature to 160°C, and the screw speed to 230 rpm. To examine the extruded products, the water absorption index, water solubility index, starch gelatinization degree, colour, phytate and oligosaccharides content were measured. For this purpose, extrudates were ground using an electrical grinder (HM- 5735) from Home Electrical Appliance and passed through a 0.25 mm sieve sized blades for some analysis, while other analyses were performed on the entire extrudates.

Physical Assessment of Extruded Puff

Finger Millet (Eleusine coracana L.), pearl Millet (Pennisetum glaucum), foxtail Millet (Setaria italic) and sorghum (Sorghum bicolour) are three selected cereals that have been evaluated in terms of physical parameters, specifically bulk density, in the context of cereals and cereal-based products. The evaluation of bulk density for the three combinations of puff products made from the selected cereals would involve measuring the mass and volume of the cereal products. The mass can be measured using a scale, while the volume can be determined by various methods such as displacement or geometric calculations. By studying and analysing the obtained results of bulk density, these conclusions can help in understanding the texture, density, and overall quality of the products, which are important factors for consumer acceptance. It’s worth noting that in addition to physical parameters like bulk density, other nutritional and sensory attributes of these cereals and their products are also crucial for evaluating their overall nutritional value and consumer appeal [20]. These may include factors such as nutrient composition, digestibility, taste, aroma, and shelf life, among others. By conducting such evaluations, can gain insights into the potential of these cereals and their products as staple foods and nutrient sources, both in developed and developing countries. Kjeldahl digestion methods (Advanced and ordinary) compared total nitrogen in non-germinated and germinated extruded puff products. Advanced Kjeldahl showed protein variations of (07.11% to 10.66%) and (09.45% to 11.98%), while ordinary Kjeldahl showed (05.11% to 10.96%) and (07.45% to 11.56%). Millet combinations revealed T3 with germinated Millets had higher protein, indicating nutritional value. Total ash, dietary fiber, and carbohydrates varied in extruded puff products. Statistical analysis indicated significant differences (p<0.0001) for each combination. Incorporating Millet and sorghum in rice flour (T3) resulted in higher protein and overall acceptable sensory properties. AOAC methods revealed significant differences (p=0.05) in proximate compositions of multi-Millet puffs with overall acceptable sensory properties [20]. This knowledge can be valuable for food scientists, policymakers, and nutritionists in promoting sustainable and nutritious food options worldwide.

Bulk Density and Expansion Ratio

Extruded products bulk density (BD) was studied by using the rapeseed displacement method and calibrated as per given equation (1) by [21]. For this purpose, the (10 ± 0.1 g) of randomly selected extruded product were weighted:

Whereas Wt. and Vep-1 are the weight in (gm) and the equivalent volume (cm3) of the extruded products, respectively. Actually, Vep is multiplication ratio between the rapeseed density (ρs) and the rapeseed weight (Ws), with same volume as the extrudates. Five replicates for each sample were prepared to study the parameters. sectional expansion index (SEI), bulk density (BD), instrumental color, and dry and bowl-life texture were evaluated. SEI and BD responses were fixed by moisture variable is incorporated; darker products were produced. Nevertheless, high together with higher temperature and lower moisture levels produces better color appearance. At high ratio desirable low hardness and crispy expanded extrudates can be generated as long as moisture is lower than about 22% [22]. The calibration of expansion ratio (ER) was according to the ratio of extruded product diameter to determine the die hole diameter of extruder with the help of caliper device, as reported by Kokseland Masatcioglu. For this purpose, ten replicates were taken for (ER) assessment.

Water Absorption Activity

The extruded products water absorption index (WAI) and the water solubility index (WSI) were determined as per equations (2) and (3) [23].

The (WSI) is the dry solid weight in the extracted supernatant, whereas (WAI) is the sediment weight without the supernatant per unit weight of the sample analyzed. Each sample was tested in a triplicate manner.

Starch Gelatinization

The starch gelatinization (SG) of the extruded products determination follows the modified method based on the procedure outlined by [24]. This method involves the formation of a blue iodine complex when amylase is released during gelatinization. Specifically, 40 mg quantity of sample was dissolved in a 50 mL of 0.15 M KOH solution and mixed thoroughly for 15 minutes. The reaction mixture was then centrifuged at 4032 x g for 10 minutes to remove out insoluble sediment. Next, 1 mL quantity of the supernatant was neutralized by the addition of 9 mL of 0.017 M HCl, and 0.1 mL of iodine reagent (1 g iodine and 4 g potassium iodine dissolved in 100 mL water). The resultant solution was mixed, and read the absorbance at 600 nm (A1) using a Cary 60 UV-VIS spectrophotometer. In addition, 1 M KOH and 0.1 M HCl solution was used to prepare control samples. The overall value of DG was calibrated by using equation (4) based on the average of three replicates.

Whereas (A1/A2) is the ratio of absorbance at 600nm of the sample to that of the control.

Phyto-Chemical Constituents of Selected Millets Puffs

The effect of extrusion processing on anti-nutrients factors namely tannin, phytate and saponin of extruded Millet-sorghum blend rice puffs puff product combinations were studied. Extrusion processing is necessary to absorb essential micro and macronutrients from added blends of puffs during extruded snacks preparations and eliminates a negative effect of anti-nutritional factors such as tannin, phytate and saponins. Thus, nutritional quality was maintained by extrusion through destruction of anti-nutritional components.

Color Determination

The CM-600d colorimeter (Konica Minolta Sensing Inc., Osaka, Japan) was used to determine the lightness (L*), redness (a*), and yellowness (b*) of both Puffs and extruded products with help of using the Spectra-Magic NX software (Konica Minolta, Tokyo, Japan). The series of experiments was replicated five times in a row.

Oligosaccharides Analysis

The presence of oligosaccharides namely verbascose, stachyose, and raffinose in Puffs and extruded products were investigated by using the HPLC with slight modifications in a method prescribed by [25]. Samples were mixed with deionized water, filtered, and separated isostatically on a cation exchange column. Identification was based on standard comparison, and quantification was based on calibration curves. Results are expressed in (mg/g dry matter) of each oligosaccharide after triplicate analysis.

Phytate Analysis

The phytate content of both Puffs and extruded products was determined by following the [26] method as its values were expressed in (mg/g) of dry matter of phytic acid. To calculate the phytate content, obtained values were multiplied by 0.282 and it is the molar ratio of phytate-phosphorus in a molecule of phytate. The analysis was done in a triplicate manner.

Sensory Evaluation

A panel of 28 semi-trained judges from University of Life Sciences and Technologies demonstrate Millet-based extruded snacks using a ranking test. Each sample is arranged in groups of three on glass plates to evaluate appearance, texture, taste, and aftertaste using an evaluation form as this form is generated through Fizz Acquisition 2.51 software. Warm black tea was used for taste neutralization between samples. The panelists ranked the samples as per most liked=1 to least liked=8 and the recorded results were calibrated as the summation of ranks for each sample (Figure 2).

Statistical Analysis

The recorded data of Millet Puffs and extruded products undergo one-way ANOVA and two-way ANOVA stats analysis, followed by Tukey’s HSD test. The two-way ANOVA stats analysis considers two major factors namely Millet puffs type and die type for further analysis. Minitab-17 ver. statistical software (Minitab, Inc., State College, PA, USA, 2010) was used to determine the significant differences among all studied parameter values at p < 0.05. The sensory evaluation data were statistically analysed by using the Friedman test with Fizz calculation (Biosystems, Cousteron, France) and a p<0.05 level of significance.

Results and Discussion

The various multi-Millet utilized to produce extruded snacks were shown in Table 1. Table 1 showed the notable differences among (L*, a*, and b*) colour patterns. Pearl Millet flour exhibited the highest a* and b* values while it had lowest L* value. In contrast, foxtail Millet flour had lightest colour, followed by finger Millet flour. Foxtail Millet flour exhibited the lowest a* and b* values, with the latter being statistically insignificant in comparison to pearl Millet flour. Additionally, Table 2 calibrated values were significantly differed (p<0.05) as the amount of anti-nutritional compounds were studied among the Puffs. Millets are known to contain several anti-nutritional compounds, including non-digestible oligosaccharides and phytic acid, which has inherent chelating characteristics to capture important divalent cations such as Fe, Zn, Ca, and Mg, which leads to lowering of availability for absorption and use in the small intestine [27,28]. However, presence of raffinose, verbascose, and stachyose oligosaccharides in human diets in an ample amount causes flatulence and discomfort in humans after consumption [29,30].

Note: *Each (n = 3) replica values were expressed as (mean ±standard deviation).

Note: *Each parameters value was expressed as (mean ± standard deviation) for (n=3).

Pearl Millet flour exhibited the highest concentration of phytates, followed by finger and foxtail Millets Puffs, respectively. Foxtail Millet flour contained the highest levels of verbascose, although it had stachyose in a lesser amount. Similarly, finger Millet flour had the highest amount of stachyose, while it has lowest amount of raffinose. However, the quantity of oligosaccharides in Millets varied as per the selected species and varieties, as well as the existing environmental conditions [31,32]. [33] reported significant variability in the amount of raffinose (4.10–10.30 mg/g), stachyose (10.70–26.7 mg/g) and verbascose (0.00–26.70 mg/g) among 18 different pea varieties. While Tahir et al. (2011) observed higher stachyose levels than raffinose and verbascose in 11 lentil varieties, as these findings were found in line with current findings. This difference in oligosaccharides level might be due to significant constraint on the extensive use of legumes at both domestic and industrial scale [34] (Figure 3).

Physico-Chemical Properties of Extruded Products

All physico-chemical parameters had significant differences (p < 0.05) among the extruded products, except water absorption index (WAI). These noticeable differences were attributed due to the type of Millet used, the type of die, and the (Millet x die) interaction, as shown in Table 3. However, it should be noted that the type of die had no significant effect on the WAI. Pearl Millet-based extruded products had the highest bulk density (BD) and water absorption index (WAI), whereas it has lowest expansion ratio (ER) and water solubility index (WSI) values. Similarly in foxtail Millet-based products had the highest (ER) and well-expanded spherical extrudates, while highest degree of starch gelatinization (SG) was recorded in finger Millet-based products. Higher ER, DG and WSI with lower BD was recorded in circular die configuration based extruded products as compared to star-shaped die. The effect of die shape on extruded product characteristics can be explained by change in friction and pressure strength. The circular die had a smaller cross-section, resulting in higher levels of friction and pressure and thus higher temperatures, which led to more expansion and lesser dense products, will obtain as it has greater ER and lower BD. On the other hand, the star-shaped cross-section had angles that could have mechanically broken bubbles in the gelatinized starchy matrix, which led to disturbance in expansion. However, with increment in the die nozzle diameter decreased radial expansion in yellow corn extrudates. Along with this, inducing higher extrusion pressure on starch gelatinization causes expansion of the extrudates products made from foxtail and finger except in pearl Millet-based products since it has higher fibre content that restricted starch gelatinization [1], [35]. Although ER and BD are important physical parameters that can influence consumer behavior in the sense of acceptability of extruded products [36].

Note: *Each parameters value was expressed as (mean±standard deviation) for (n=3).

Earlier studies by [37] who reported an inverse relation between BD and ER, as observed in this work, where a negative correlation was found (r = -0.631; p = 0.093). The presence of huge number of fibres in the feed Puffs can also affect the ER and BD values by reducing overall expansion on account of cell-wall rupture, resulting in a compact and hard product with an undesirable texture. In this context, Red lentil flour, with the highest fibre content, led to extrudates with the lowest ER and highest BD indicating the importance of considering fibre content while developing extruded product as described by [38]. The extrusion-cooking conditions, including the die used, may also impact the WAI and WSI values, which represent the amount of water that can be absorbed by the extruded product and the quantity of soluble substances formed during the extrusion process from starch, proteins, and fibres. The presence of fibres in higher amount could also influence functional properties [39]. Instead of this, WSI was also influenced by other factors such as legume type, die shape, and legume-die interaction, while effect of die shape had no significant effect on WAI. Higher fibre levels in the legume led to an increase in WAI, as they absorbed and retained water within a well-developed starch-protein-polysaccharide network, as discussed previously by [40].

Extruded Products Color

Colour is a crucial aspect of food products that can significantly affect consumer acceptability. Extrusion-cooking affects the colour features of the products, making them darker than the Puffs. The colour components of the extrudates were influenced by Millet type, die configuration and their interaction. The observed colour features were attributed to existing pigments and the Maillard reaction occurring during extrusion-cooking (Table 4). Star-shaped extrudates had greater L* and lower a* values (except for pearl Millet) compared to spherical shape extrudates, while b* index had found non-uniform trend. This fluctuating trend is due to pigment degradation as temperature rises and shear stress during extrusion is responsible to alter color, especially for carotenoids. The decrement in L* and increment in a* values may be linked to the melanoidins formation during the Maillard reaction, while increment in b* may be results from the formation of yellowish compounds during the initial stages of the Maillard reaction or from lipid oxidation. An advantage of larger die cross-section reduces extrusion pressure and heat, leading to a less intense Maillard reaction and reduces browning and flavor development [41]. Thus, star-shaped extrudates were obtained with the help of larger cross-sectional die and a less drastic extrusion process involved to obtain lighter colour than spherical shaped extrudates [42].

Note: *Each parameters values were expressed as (mean± standard deviation; n = 3); indicating the significant differences (p < 0.05) among the sphere and star shaped products considering the interaction between the Millet and die.

Anti-Nutritional Component of Puffs and Extruded Products

The extrusion-cooking process and the type of raw material used can influence the levels of anti-nutritional compounds in legume extrudates [43,31]. A comparison of the native Puffs shown in Table 2 and the extrudates in Table 4 revealed different kind of behaviour for various anti-nutritional compounds. In present study the phytates content decreased as the extrusion-cooking of finger Millet (12% on average) and foxtail Millet (7.9% on average) Puffs initiated for both star-shaped and spherical products, possibly on account of thermal processing that is associated with the extrusion-cooking process. [12] found that total phytates were more greatly reduced in lentil flour extruded at 160°C compared to 140°C. However, oligosaccharides, particularly stachyose and raffinose increased during extrusion cooking. This could be attributed to the high temperature and pressure involved in extrusion-cooking, which break the bonding between oligosaccharides and other macromolecules, or alter the food matrix structure, leading to better extractability of anti-nutritional compounds [6]. Similar kind of results were observed in pea-rice gluten-free expanded products and extruded lentil snacks by other researchers [6,25]. It was observed that there were differences in verbascose, stachyose and raffinose content in Millets extrudates just because of Millet type, die configuration and their interaction, but their phytic acid was not affected by die shape. In case of pearl Millet based spherical extrudates have higher verbascose content than starshaped, while raffinose content increased to 7% in the latter. Common bean-based star extrudates had higher stachyose and raffinose content compared to spheres made from the same flour, which decreased by 1.7% and 7.5%, respectively [23]. Extrusion conditions and legume type affected oligosaccharide behaviour, with a higher temperature and pressure increasing stachyose and raffinose contents. An obtained results showed that oligosaccharides content was found higher in spherical shaped products than the star-shaped after die inducing higher pressure and heat generation, particularly for raffinose [39].

Sensory Evaluation of the Extruded Products

The sensory attributes like appearance, texture, taste, and aftertaste of the extruded products were significantly (p <0.05) affected by the type of flour and die used. The products ranking is decided as per test results (Table 5). As per sensory attributes the star-shaped extrudates prepared from pearl Millet were found to be least preferred acquiring lowest rank sum for “texture” due to their hard structure and difficult to chew, bland taste, and aftertaste. However, pearl Millet based spherical extrudates were liked for “appearance” and “aftertaste”, similarly notified in spherical and star-shaped extrudates from finger and foxtail Millets. Previously [10] studies showed that high values of BD and hardness can produce undesirable products for consumers. In contrast, common Millet extrudates, particularly a spherical one, were preferred in terms of all considered attributes owing to their properly puffed and crunchy nature, pleasant taste and aftertaste as discussed by [28]. In another study, extrudates made from blends of red lentil and corn were found to be more accepted than including (100%) lentil extrudates [17]. In general, the spherical shaped extruded products were favored as compared to star-shaped ones, indicating that preferred shape plays a crucial role in consumer perception and acceptability of food products. This is supported by studies that suggest that shape can even affect taste perception. However, there was no significant difference observed between spherical and star-shaped extrudates in terms of taste and aftertaste, possibly because textural features, such as appearance and structure, had a greater impact.

Note: *Each parameters values were expressed as (mean± standard deviation; n = 3).

Conclusion

The study aimed to understand how the die configuration affects the extrusion of Millets. Most industry dies have a circular cross section, but the effect of a star-shaped die on Millet was still unknown. Obtain results of the study showed that the die-hole diameter significantly affects the physicochemical properties and sensory qualities of the extruded snacks. The use of a star-shaped die to produce products with a lower ER and higher BD than spherical extrudates and it is preferred one on account of lowering the friction resistance during extrusion. The increased knowledge on die configuration could aid in the expansion of Millet-based raw materials at maximal level to meet consumer satisfaction for healthy and palatable food products.

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Journals on Medical Informatics

Nigella Sativa use for the Treatment of Cancer Tasawar

Introduction

Cancer is a big problem in society today and is the second most common cause of death after heart attacks. Every year, many people die from different types of cancer, even though we try really hard to find ways to stop and treat it. In the last 100 years, modern medicine has made big progress in treating diseases. But a lot of illnesses, like different types of cancer, are still not completely treated. Researchers are looking at both old and new ways of healing to find new and effective treatments [1]. Nigella sativa has been used as medicine for a long time. This tradition started in Southeast Asia and then was used in ancient Egypt, Greece, the Middle East, and Africa. The Islamic tradition and the healing power of medicine are important for healing and are an unusual type of medical treatment [2]. This plant is a flower plant and its seeds are used as a spice in cooking. In English, the seed is often called black cumin, and in ancient Latin it was named “Panacea”, showing that it was used for healing. In Arabic, the seed is called “Habbah Sawda” or “Habbat el Baraka”, which means “Seed of blessing”. In Arabic, people call the seed “Habbah Sawda” or “Habbat el Baraka”, which means “Seed of blessing”. This tree is called “Kalo jeera” in Bangladesh, “Kalonji” in India, and “Hak Jung Chou” in China [3]. The plant’s seeds and oils are important for medicine. The main parts of N. Sativa could help keep you healthy and might treat different illnesses like cancer. One way it works well is by lowering the chance of atherosclerosis. It does this by lowering bad cholesterol in the blood and raising good cholesterol. It helps diabetes by making the body healthier and protecting the cells that make insulin in the pancreas. This can help as a treatment for diabetes. Take care of and keep safe. It helps control high blood pressure [4]. It effectively reduces airway inflammation in people with asthma, and its components show potential as a complementary treatment for schistosomiasis. In addition, its oil also protects kidney tissues from damage caused by harmful oxygen molecules, thereby preventing kidney dysfunction and structural abnormalities. For countless centuries, seeds, oils and extracts derived from N. sativa have been used for their anti-cancer properties in traditional systems of medicine such as Unani, Ayurveda and ancient Chinese medicine. These systems were developed in Arabic, Indo-Bangla and Chinese respectively [5].

Role of sativa as Anti-Cancer Mediators

A lot of useful substances have been found in the seeds of N. Sativa is a type of cannabis plant. Instead, it should be written in a simpler and easier to understand language. Nsativa seeds contain oils, proteins, alkaloids and saponins. These components were analyzed to quantify four important pharmacological elements in the oil: thymoquinone, dithymoquinone, thymohydroquinone and thymol. Nsativa seeds are seeds of the sativa plant. The main component in the essential and fixed oils of the seeds, called thymoquinone, is believed to be responsible for a significant portion of their biological effects. Thymoquinone is often recognized for its powerful abilities as an antioxidant, anticancer, and antimutagenic agent. In addition, thymoquinone has acceptable safety levels, especially when administered orally to animals in experiments. Alpha-hederin is a compound found in black seed that comes from the seeds of the N. sativa plant [6].

Blood Cancer

Thymoquinone stops the growth of a type of cancer cells called HL-60 cells, which are found in human myeloid leukemia. Researchers studied different forms of thymoquinone with 6-alkyl residues and terpenes in HL-60 cells and 518A2 melanoma. The scientists discovered that these substances can cause a process called apoptosis, which is when the DNA breaks into pieces, the energy in the cell decreases, and there is a small increase in harmful chemicals. They found that α-hederin made P388 murine leukemia cells die by causing more apoptosis to happen, and this happened more as the dose of α-hederin and the time went up [7].

Breast Cancer

The mix of alfalfa, melatonin and retinoic acid helped lessen the bad effects of DMBA on breast cancer in mice. Thymoquinone was tried on breast cancer cells called MCF-7/Topo. Thymoquinone has a special part called terpene and 6-alkyl. They discovered that these substances made cells die through a process called apoptosis [7].

Colon Cancer

Thymoquinone can help fight colon cancer cells and promote cell death. This was shown in a study where the N.sativa volatile oil was used on colon cancer cell line HCT116. N.sativa can stop the growth of colon cancer in rats, after it has started, without causing any noticeable negative effects. Thymoquinone is a substance that can be used as a treatment for colon cancer cells. It works similarly to a drug called 5-flurouracil. However, thymoquinone did not have any effect on HT-29 (colon adenocarcinoma) cells [1].

Pancreatic Cancer

Thymoquinone is the most important chemical in Nigella sativa, which is known for its important healing properties. The study looked at sativa oil extract affects the way pancreatic cancer cells grow and die. Scientists have suggested using thymoquinone to stop inflammation and encourage cells to die in a certain way [8]. This could be a new way to treat inflammation. Thymoquinone can help make pancreatic tumors more sensitive to standard treatments by reducing the effects of gemcitabine or oxaliplatin on NF-kappa B activation. Mucin 4, a big molecule with sugar attached to it, is not working properly in pancreatic cancer. This strange expression is important for many things like cells change, grow, spread, and resist chemotherapy in pancreatic cancer. The study looked at thymoquinone affects MUC4 expression in pancreatic cancer cells [7].

Hepatic Cancer

A lot of research has been done to study well different treatments can kill cancer cells. This study wanted to see if sativa seeds can affect liver cancer cells grow. The test showed that N can stop HepG2 cells by 88% after being left with them for 24 hours at different amounts. So, using sativa extract is very important in academic discussions. Thymoquinone has been found to help the body’s quinone reductase and glutathione transferase work better when taken by mouth. The properties of thymoquinone make it possible to prevent cancer caused by chemicals and protect the liver from damage [9].

Lung Cancer

The cancer-fighting ability of α-hederin found in N.staiva is being studied. The research looks at sativa affects lung cancer in mice. A study also showed that putting honey and N. Adding sativa to the diet makes a big difference. Sativa helps protect against oxidative stress and inflammation caused by certain chemicals and can help prevent lung, skin, and colon cancer. Alfalfa has been looked at a lot because it might have healing powers [2]. These chemicals have different effects on living things, such as reducing inflammation, fighting cancer, and preventing damage caused by harmful substances. Additionally, they look like they could help treat heart problems and might assist in cancer treatment. Understanding α-hederin and thymoquinone work can help us use them in medicine. More research is needed to understand they can help people and make sure they are safe and work well in medical treatment. N sativa does not have a big impact on cell death or programmed cell death in lung and laryngeal cancer cells [10].

Skin Cancer

Topical use of N. sativa is a common form of management used in a variety of settings. sativa extract showed an inhibitory effect on the initiation and promotion stages of skin carcinogenesis in mice when administered intraperitoneally. Skin application of 20-methylcholanthrene resulted in a significant reduction in soft tissue sarcomas, which was limited to 33. 3%, compared to 100% incidence observed in the MCA control group [11].

Fibrosarcoma

Thymoquinone, taken from the seeds of Nigella sativa, has been studied a lot for its possible healing powers because of its different biological effects. The administration of sativa one week prior to and following MCA treatment exhibited a notable hindrance in the development of fibrosarcoma tumor occurrences, as well as a reduction in tumor mass, by 43% and 34% respectively, in comparison to the outcomes observed in the MCA solitary treatment group. Furthermore, it was observed that thymoquinone exhibited a delayed onset of fibrosarcoma tumors induced by the administration of MCA. Moreover, in vitro investigations demonstrated that thymoquinone exhibited inhibitory effects on the viability of fibrosarcoma cells. Alfalfa oil is a special kind of oil from the Medicago plant that can decrease the ability of human fibrosarcoma cells to break down fibrin in lab tests [1].

Renal Cancer

There is an existing body of research that highlights the potential chemo-preventive efficacy of N.sativa, a substance that has garnered significant scientific interest. N sativa has inhibitory effects on renal oxidative stress, hyperproliferative response, and iron nitrilotriacetate-induced renal carcinogenesis. In this experimental study, alfalfa was administered orally to rats for therapeutic purposes.The administration of sativa elicited a pronounced reduction in the generation of H2O2, synthesis of DNA, and occurrence of tumors [12].

Prostate Cancer

Thymoquinone comes from Nigella seeds and has many important medical qualities. The extract from the sativa plant can slow down the making of DNA, stop cells from growing, and reduce the ability of cancer cells in the prostate to survive. These changes were seen only in cells that have cancer, not in cells that do not have cancer. It was found that this result happened because the androgen receptor and transcription factor were decreased [13]. Thymoquinone was found to work well in treating both hormone sensitive and hormone-refractory prostate cancer in different experiments. Research done in the lab and in animals shows that thymoquinone can stop the growth of new blood vessels. In addition, thymoquinone was found to stop the growth of blood vessels in a human prostate cancer model in mice. Also, when used in small amounts, thymoquinone stops the growth of human prostate tumors, with very few chemical side effects. Furthermore, thymoquinone affects endothelial cells more than cancer cells by causing cell death, stopping cell growth, and blocking cell movement. Thymoquinone stopped the activation of a protein called extracellular signal-regulated kinase, which is usually turned on by a substance called vascular endothelial growth factor. However, it did not stop the activation of the vascular endothelial growth factor receptor 2 [14].

Cervical Cancer

The extracts of N. sativa were obtained using methanol, n-hexane, and chloroform. The sativa plant caused human cervical cancer cells to die. “We looked at terpene-terminated 6-alkylthymoquinone residues affect cervical cancer cells that are resistant to multiple drugs. ” Thymoquinone derivatives have been discovered to make cells die in a certain way called apoptosis [4].

Molecular Mechanism of sativa Action Against Cancer

Cancer happens when cells grow in an unusual way because of changes in the genes. So, any medicine that fights cancer can either protect DNA from changing or kill cancer cells that have changed. N.sativa seeds have been extensively studied for their pharmacological properties. Thymoquinone, a major active compound found in N. Sativa seeds, has demonstrated promising therapeutic effects in numerous medical conditions. These studies have shed light on the potential of thymoquinone as a valuable agent in disease prevention and treatment. Sativa exhibits its efficacy in combatting cancer cells through various molecular pathways. Possible mechanisms underlying the action of thymoquinone. Thymoquinone helps kill cancer cells by turning on genes that cause cell death and turning off genes that keep the cells alive. Thymoquinone effectively hinders the activation of Akt by means of dephosphorylation, ultimately impeding the viability of cancerous cells [15]. There is currently ongoing research regarding topic N.sativa in the academic community. Nsativa or thymoquinone oil exhibits antioxidant properties, leading to enhanced enzymatic activity of antioxidant enzymes including superoxide dismutase, catalase and glutathione peroxidase. Notably, increased activity of these antioxidant enzymes has been shown to be beneficial in fighting various forms of cancer. The use of alfalfa oil or thymoquinone has been observed to reduce the toxicity of various anticancer drugs due to enhanced activation of antioxidant mechanisms. This result suggests that these drugs have great promise for clinical use in mitigating the harmful effects associated with anticancer drugs [4].

Concluding rrmarkers

Nigella sativa (N.sativa), commonly known as dim seeds, have been broadly inspected for their promising anti-cancer properties. Many in vitro and in vivo tests have outlined the practicality of N.sativa in preventing the improvement and development of diverse sorts of cancer cells. In extension to its facilitate cytotoxic impacts, N.sativa has been found to have solid antioxidant, anti-inflammatory, and immunomodulatory properties, all of which contribute to its anti-cancer development. Besides, N.sativa has appeared potential in sensitizing cancer cells to customary chemotherapy and diminishing chemotherapy-induced side impacts [16].

Nigella Sativa Good for Cancer

Nigella sativa has attracted a lot of interest from researchers and scientists. Extracts and seeds of the N. sativa plant and its active ingredient thymoquinone have been thoroughly studied, with excellent results showing that N. sativa has medicinal potential. The sativa strain tends to have medicinal properties that can be effective in treating a variety of ailments, including cancer [7].

Sativa Extracts be used to Treat Cancer

N.sativa extracts have the prospective application in the advancement of efficacious therapeutic agents for combating cancer. These fractions can act alone or in combination with chemotherapeutic drugs that have proven to be effective agents for modulating tumor initiation, proliferation and metastasis, making them possible treatments for many types of cancer [17].

Pro-Apoptotic and Anti-Proliferative Effects of Sativa

N.sativa has the ability to fight against cancer. Researchers have gathered a lot of evidence about sativa by studying it outside of living organisms and inside them. They have used different types of cells and animals to do this. The scientists in the study said that they didn’t look at the extracts from each individual plant in the mixture could fight cancer because only the mixture is used in cancer treatment. Sativa extracts are extracts from the sativa plant. In an initial test on living organisms, we applied N.sativa using a cream or ointment on the surface. The N.sativa extract slowed down the development of skin cancer and reduced the appearance in mice when they were exposed to certain chemicals [18].

Signaling Pathways Fundamental the Anti-Cancer Effects of Sativa

Many tests were done in the lab and on living organisms to understand N.sativa fights against cancer at a molecular and cellular level. Sativa is a type of plant. The main ways that N.sativa (a specific substance) helps fight against cancer are not yet fully understood, but they have been well-documented. The effects of sativa are mostly due to their capability to control the action of important enzymes. Reduce swelling and encourage the natural death of cancer cells [15].

Sativa Phytoconstituents and Anti-Cancer Effects

The anti-cancer impacts of N.sativa are exceptionally critical. The most fixing in N.sativa, called thymoquinone, has been connected to its impacts. Thymoquinone has been found to have a few useful impacts on cancer cells. It can offer assistance halt their development, empower cell passing, secure against harm caused by substances called oxidants, decrease the probability of changes, anticipate the arrangement of modern blood vessels that tumors got to develop, and moderate down the spread of cancer cells to other parts of the body. This N.sativa herb has been appeared to have properties that can battle cancer and murder cells. Be that as it may, we do not completely get it it works however, so more investigate is needed to figure out the points of interest. Sativa phytoconstituents are the common compounds found within the sativa plant (Figure 1) [4].

Figure 1

Conclusion

Nsativa is a very popular herb that has been used by people for a long time. Many people think Nsativa is a special plant that can help heal and reduce the effects of infections, such as cancer. The ability of N.staiva to fight cancer. Sativa is good because it can help stop cells from growing, help cells die, and protect cells from damage and cancer. N. sativa ability to defend effectively. Sativa can help stop tumors from forming and spreading, partly because it can prevent them from getting worse and has a mild stimulating effect that is safe. The tests done in the lab and on living organisms show that N. Sativa extract can be used to create helpful and powerful starting materials that can be used at different times during the process. Cancer can form tumors in different parts of the body. There are different treatments for different types of cancer. More testing is needed to understand N helps fight cancer at the atomic and cellular level. Sativa researchers want to understand the exact ways that N. stops certain signals in the body. The sativa extract is involved in the development of tumors and cancer. In the future, we need to study N.staiva can work together with other things to fight cancer. Sativa extract being used to prevent and treat cancer in research and medical settings.

For More Articles: Biomedical Journal Impact Factor: https://biomedres.us

Journals on Cancer Medicine

Nigella Sativa use for the Treatment of Cancer Tasawar

Introduction

Role of sativa as Anti-Cancer Mediators

Blood Cancer

Breast Cancer

Colon Cancer

Pancreatic Cancer

Hepatic Cancer

Lung Cancer

Skin Cancer

Fibrosarcoma

Renal Cancer

Prostate Cancer

Cervical Cancer

Molecular Mechanism of sativa Action Against Cancer

Concluding rrmarkers

Nigella Sativa Good for Cancer

Sativa Extracts be used to Treat Cancer

Pro-Apoptotic and Anti-Proliferative Effects of Sativa

Signaling Pathways Fundamental the Anti-Cancer Effects of Sativa

Sativa Phytoconstituents and Anti-Cancer Effects

Figure 1

Conclusion

For More Articles: Biomedical Journal Impact Factor: https://biomedres.us

Open Access Clinical and Medical Journal

Effect of Caffeine, Gender, and Time of Day on Working Memory in Medical Students

Numerous studies have looked at psychopharmacological and electrophysiological effects of caffeine on the human brain and heart [1-4]. Effects of caffeine in humans have been found to enhance mental performance, alertness, and mood especially when subjects are fatigued [5-7]. However, many variations exist in literature among the subjects assessed and results achieved. Firstly, some studies show that, in free recall tasks, caffeine either has no effect [8-11], a beneficial effect on cognition and memory [12-14] or can impair performance in tasks involving working memory8. Studies have attributed these differences in cognition to variations of mental performance in response to caffeine between the methods used in testing short term memory, the amount of caffeine given, sex and age of participants as well as the time of day for testing [15,16]. First, when testing the effect of caffeine on short-term memory, a discrepancy exists among research studies. In 1958, Kirchner conducted an experiment to measure very short-term retention in younger and older Ss utilizing a visual display involving a rapidly moving light; the results indicated that older Ss slumped in performance, make more errors of omission and more random responses much sooner than younger Ss, in both relative and absolute terms; thus indicating the inability to organize incoming and outgoing information as rapidly as the younger Ss [17]. Since then, other studies in the past have used multiple variations of memory tasks, causing a large percentage of previous data on short-term memory to be incomparable [18-20]. Repovs and Baddeley have established a working theory for a multi-component model of working memory [21].

They define working memory as being composed of three aspects: a phonological loop, visuospatial sketchpad, and a multimodal store, the episodic buffer which allows integration of short term and long-term memory. This article proposes the testing of all aspects of short-term memory defined by Repovs and Baddeley [21], through the integration of a visual, auditory, and combined audio and visual digit span test, which has been established as one of the most accurate tests in predicting working memory progress [22]. Second, a wide range of research has been performed to determine a concentration of caffeine for studies that best mimic natural caffeine levels consumed by most of the regular population. Cheng and colleagues have determined that caffeine levels between 70mg to 300mg show a significant decrease in the clearance of caffeine as the dose increases; thus, indicating saturable caffeine metabolism in the dose range tested [23]. Also, around 150mg-200mg of caffeine has been found to imitate moderate caffeine intake by subjects, the majority of whom consume around three cups of coffee per day [24]. This study, therefore, proposes to use 200mg of caffeine, in a one-tablet dose, which best reenacts natural conditions for subjects. To achieve the full influence of the caffeine dose, it is also proposed that subjects wait 30 minutes, since between 15 minutes to 120 minutes caffeine shows an effect in humans [25].

Additionally, studies in the past have proven that sex and age of participants have varying results, with females and males performing at different scales from each other depending on the amount of caffeine given, time of the study, and type of test used [26]. Furthermore, differences in the age of participants have also shown variable results, with subjects over the age of fifty found to be more sensitive to the objective effects of caffeine when compared with individuals below 50 years of age [9]. Due to such variation in participant qualities, this study proposed to maintain a separate comparison between the effects of caffeine in males and females and to use subjects below 50 years of age, to eliminate discrepancies and allow for more accurate comparisons between groups. Lastly, as discussed by Ryan and colleagues, caffeine can have a different effect on individuals if the memory task is performed in the morning versus the afternoon [14]. The performance depends on numerous obstacles, such as whether the participant is a morning person or whether their natural decline in function that accompanies the afternoon overlies the time frame for the mental assessment. It is thus proposed for this research that we conduct two trials, one in the morning and one in the afternoon to account for individual differences in performance on the memory tests.

Methods

Study Design

We conducted two randomized double-blind trials lasting one week in duration (one in the morning and one in the afternoon) involving healthy participants at the St. James School of Medicine. Each participant consented to participate and was compensated for participation. The study protocol was approved by the institutional review board. The authors vouch for the fidelity of this report and the accuracy and completeness of the data and the analyses.

Study Procedure

Before the first round of testing, each participant completed a survey recording their age, sex, race, the average duration of sleep per night, and average caffeine use in a normal week. Eighty-one participants (40 in the morning trial and 41 in the afternoon trial) were randomly assigned into groups receiving caffeine (one 4 everfit 200mg tablets) and groups receiving placebo (one 400IU tablet of Nature’s Bounty vitamin D). Each day during the study, the participants were given a pill with water then asked to wait for a half-hour to allow the contents to be absorbed. After the half-hour waiting period, each subject was administered three-digit span tests (auditory stimulus, visual stimulus, and both auditory and visual stimuli) via a computer program design for this study. This process was repeated every day for the duration of the study. Their results were recorded by the program and analyzed at the end of the study using Microsoft Excel.

Program Specifics

A program was created to present the digit span test to the participants; results were collected and maintained consistently with the use of Microsoft Visual Studio 2010 Visual Basic Net and Microsoft Excel software. The software runs using Windows 7 32-bit operating system. The volume of the PC is set by the user before the trial began and is not controlled by the program other than to either use or not to use an auditory presentation. The visual presentation of the integers is displayed in a black background and a white foreground with a 14-point Times New Roman font. The window completely covered the screen of the monitor to reduce distractions. The generated number is displayed in the center of the screen for one second and removed before the next number is generated and presented. When an auditory presentation is selected, a soft feminine voice announces the generated number in English with the annunciation of each number lasting one second. If both an auditory and visual presentation is selected the number is displayed and announced at the same time. Each number in the series is presented in the same manner for the duration of the trial. Each integer is presented separately in one-second intervals beginning with a minimum series of three integers for three seconds. After the last integer is presented, the program clears the screen and displays a new screen with instructions for the subject to enter the numbers just revealed into the response box in the order in which the integers were presented. There is no time limit to enter the values into the response box.

The test subject must press the enter key when the last number is entered into the response box. The program will store in a database the series of integers presented, the test subject’s responses, and the results of the comparison of the generated series to the test subject’s entered values. If the response is the same as the generated series the program will increase the series of integers to present to the subject in the next round by one integer. If the response differs, the program will in the next series give the test subject another attempt at the current number of generated integers in a series (up to a maximum of two attempts). After the second attempt at the designated number of integers in a series, the program decreases the series length by one integer until the minimum number of integers to display to the test subject in the series is reached (currently the minimum series length is three integers). The program will present a total of fourteen series of numbers and after each series, it determines and store the results for that test subject. After each completed trail the total number of correct responses and the maximum length of integers presented for a correct response is recorded for each test subject.

Statistical Analysis

We estimated that our sample of eighty-one participants would provide 95% power to detect a significant difference for the primary endpoint at a two-sided significance level of 0.05, assuming a normal distribution.

Analyses were performed on each data set and differences between the treatment groups were evaluated with the use of a two-tailed unpaired uneven variant student t-test. All reported P values are two-sided; a P-value of 0.05 or less was considered to indicate statistical significance. All analysis was done using Microsoft Excel.

Results

(Table 1 & Figure 1) shows the mean raw scores for digit span tests and the mean longest recorded test score with p-value, respectively. (Figures 2 & 3) show the graphic presentation of caffeine trial in the morning and afternoon, respectively. (Figure 4) shows a graphical presentation of the time frame and gender against p-value. (Figures 5-8) show the Histograms demonstrating the frequency of the longest recorded scores for caffeinated and placebo males and females in the morning and afternoon sessions, respectively.

Morning Trial

No significant difference was observed in male test scores for both the caffeine (Audio vs. Visual p=0.645, Audio vs. Audio/Visual p= 0.279, Visual vs. Audio/Visual p=0.537) and placebo (Audio vs. Visual p=1, Audio vs. Audio/Visual p=0.709, Visual vs. Audio/Visual p=0.709) groups. No significant difference was observed in female test scores for both the caffeine (Audio vs. Visual p=0.510, Audio vs. Audio/Visual p=0.402, Visual vs. Audio/Visual p=0.144) and placebo (Audio vs. Visual p=0.922, Audio vs. Audio/Visual p=0.638, Visual vs. Audio/Visual p=0.562) groups. 200mg dose of caffeine had no effect on male test scores when compared to the test scores of males taking the placebo (Audio p=0.478, Visual p=0.690, Audio/Visual p=0.613). Female subjects taking placebo recorded significantly longer scores across each of the three tests as compared to the test scores of caffeine exposed females (Audio p=5.63×10^-4, Visual p= 3.02×10^-3, Audio/Visual p=0.019). Caffeinated males performed significantly better during the individual audio and visual tests than caffeinated females (Audio p=0.006, Visual p=8.37×10^-3). The combined audio/visual test showed no significant difference between males and females (Audio/Visual p=0.052). No significant difference was noted during any of the tests between males and females exposed to the placebo (Audio p=0.954, Visual p=0.988, Audio/Visual p=1.00).

Afternoon Trial

Both caffeinated and placebo males performed significantly better on audio tests as compared with visual tests (p= 1.49×10-3, p=1.31×10^-3respectively), while placebo males also performed significantly better on audio/visual combination tests as compared with visual tests (p=4.55×10^-3). No significant difference was observed in any other test for caffeinated (Audio vs. Audio/Visual p=0.119, Visual vs. Audio/Visual p=0.088) or placebo (Audio vs. Audio/Visual p=0.769) males. No significant difference was observed in the placebo or caffeinated females when comparing audio and audio/visual combination tests (p=0.509 and p=0.211 respectively). However, significant differences were noted between all other tests for both placeboes (Audio vs. Visual p=6.31×10^-3 and Visual vs. Combined p=2.78×10^-2) and caffeinated (Audio vs. Visual p=3.39×10^-4 and Visual vs. Combined p=1.43×10^-2) females. In each case, females recorded a lower mean score for visual tests than either the auditory or auditory/visual tests. 200mg dose of caffeine had no effect on male test scores when compared to the test scores of males taking the placebo during individual audio and visual tests (Audio p=0.352, Visual p=0.170). Placebo males performed significantly better than males on caffeine during the combined audio-visual test (Audio/Visual p=2.58×10^-2). Females taking 200mg dose of caffeine had no significant difference in test scores when compared to females on placebo in all tests (Audio p=0.934, Visual p=0.546, Audio/Visual p=0.498). Males taking placebo performed significantly better than females taking placebo (Audio p=3.42×10^-2, Visual p=4.73×10^-2, Audio/Visual p=1.21×10^-2). No significant difference was seen between males taking caffeine and females taking caffeine (Audio p=0.224, Visual p=0.216, Audio/Visual p=0.409).

Morning vs. Afternoon

When comparing males from the morning session to males in the afternoon session no significant difference was noted in test scores for males exposed to caffeine during audio tests (Audio p=0.379), or males taking the placebo during audio and combined audio/visual tests, (Audio p=0.471, Audio/Visual p=0.126). Males in the caffeine group performed significantly better in the morning than the afternoon during visual and combined audio/visual tests (Visual p=9.92×10^-6, Audio/Visual p=3.87×10^-4). Males taking placebo in the morning also performed better during visual tests than males taking placebo in the afternoon (p=7.31×10^-3). When comparing females from the morning to the afternoon no significant difference was seen in females taking caffeine during the audio tests (p=0.355). Females taking placebo performed significantly better in the morning in all tests (Audio p=4.07×10^-3, Visual p=3.09×10^-7, Audio/Visual p=3.97×10^-5). Females also performed significantly better in the morning as compared to the afternoon when taking caffeine in visual and combined audio-visual tests (Visual p=2.33×10^-3, Audio/Visual p=1.04×10^-2).

Discussion

The effect that caffeine has on memory functions in humans has been noted in numerous separate studies that, perplexingly, arrive at differing and sometimes opposed conclusions. The truth is that the end effect of this compound’s ability to add or detract from mental functioning is dependent on the various parameters of these experiments that most, if not all, of these conclusions, are correct. More importantly, not only can the results be supported by the various data, but the differing of the conclusions can be explained. An anecdotal conclusion is that caffeine has been shown to both improve and impair working memory. There is a plethora of variables, many interdependent, that make quantifying working memory often an almost insurmountable task. Parameters include weight, gender, hormone levels, time of day, fatigue, and age. To begin, according to the U.S. Department of Health and Human Services, the average U.S. woman is 5′ 3.7″ (162 centimeters) tall and weighs 152 pounds (69 kilograms). This is with respect to a Body Mass Index of 26.3 kilograms/meters², which is slightly less than the average men. The average U.S. male stands 5′ 9.1″ (175 centimeters) tall and weighs 180 pounds (82 kilograms), with a Body Mass Index of 26.5 kg/m². Therefore, on the average, the same standard dose provides at onset 18.42% more caffeine per unit of body mass in females than it does to males. As our study looked at the difference in male and female immediate responses to caffeine, it did not account for weight variances between subjects.

In other words, equal concentrations had been given in this study to determine the instant response to caffeine, without noting that weight variance between participants could have had a fast-immediate effect on the study results. It is therefore possible that, given on average the lower weights of female participants as compared to male participants, there could have been a difference in mg of caffeine / kg, which potentially contributed to the statistically significant memory impairment seen in the caffeinated females but the precariously absent effect in the caffeinated males. Differences between males and females regarding caffeine intake can also be attributed to the metabolism of caffeine, normally executed by cytochrome P450 1A2 (CYP1A2) enzyme. In females, numerous studies have hypothesized that estrogen might play a role in the metabolism of caffeine. Pollock and colleagues assessed the effects of exogenous estrogen on caffeine metabolism [27]. They placed women on eight weeks of estrogen replacement and measured their caffeine metabolic ratios (CMR) before and after the therapy. For all participants, CMR levels were decreased after the estrogen therapy as opposed to before beginning the therapy, which suggests that estrogen might hinder the metabolism of caffeine by CYP1A2. Similar results were demonstrated by Hong and colleagues, where CMR in premenopausal and postmenopausal women was decreased by 22% and 15% with high levels of free circulating estradiol as opposed to women with low levels of free circulating estradiol [28].

CYP1A2 was also negatively associated with the percentage of circulating free estradiol. Thus, estrogen could influence CYP1A2, and further decrease the metabolism of caffeine by the enzyme. This connection between the different metabolism of caffeine in males and females can likewise be supported by Scandlyn and colleague, who shows that there exists a sex-specific difference between CYP1A2 activity in males and females, with males having a higher enzyme activity than females [29]. Since CYP1A2 metabolizes caffeine, higher levels of the enzyme in males and a more suppressed enzyme under the influence of estrogen in females can explain the difference in sex in performance on mental tasks. This is especially important when recalling that high doses of caffeine can negatively impact working memory [15]. These factors could provide an alternative or compounding reason for the difference between performance in males and females on our digit span test. Estrogen is not the only hormone that could contribute to altered working memory. It is known that human physiology is influenced by natural circadian rhythms in which an increase in alertness occurs in the early morning and declines throughout the day demonstrating low levels of alertness in the early afternoon independent of food intake [30]. In our study, placebo males performed equally well in the morning and afternoon in auditory and combination tests. However, during the visual test, men recalled longer strings of numbers in the morning relative to the afternoon.

Interestingly, females in the placebo group performed significantly better in the morning than in the afternoon in each test. These results indicate some time-of-day effects where male’s visuospatial memory and female’s visuospatial and phonological loop may be more prone to inhibitory effects that occur during a day. This observed decline may be associated with the maximal concentrations of glucocorticoids found in the blood in the morning, which follow alertness by declining levels as the day continues. According to Lupien and colleagues working memory tasks decreased signiﬁcantly under the highest dose of hydrocortisone [31]. Additionally, curve ﬁt estimations exhibit a “U-shaped” relationship between working memory task performance and changes in glucocorticoid levels after hydrocortisone infusion. Furthermore, there is an upper limit to the point at which caffeine can provide enhanced mental capabilities. This upper limit of how well you can perform is in the typical case: if you are well-rested and awake then you are already at the upper limit of your functioning capabilities; thus the caffeine probably does not result in enhanced performance; but paradoxically, it probably lowers your performance because it makes you ‘jittery’ and being ‘too aware’ because of heightened senses which reduce your ability to focus. So, the benefits of caffeine are greatest neither for those test subjects that are well rested nor for those in total exhaustion, but for the mildly exhausted or fatigued subject [32].

As such, both cortisol levels and fatigue could have contributed to the differences observed between the morning and afternoon groups. Finally, it has been seen in past research that varying amounts of caffeine have a positive or negative effect on tasks testing working memory. For example, Kaplan and colleagues have concluded that caffeine in low doses enriches working memory, while at high doses caffeine impairs it [15]. Furthermore, Rogers in his study has found that the effects of varying concentrations of caffeine show different results depending on the fatigue or sleep deprivation, with low doses of caffeine increasing alertness while high doses increase both alertness and performance on tasks testing working memory [33]. Our study demonstrates caffeine’s potential ability for having an enhancing effect on an aspect of auditory processing and/or rehearsal in working memory. Both caffeinated males and females recalled significantly fewer digits in visual and combination tests from morning to afternoon while being conserved for the auditory tests. A similar result was seen in a study in which a group of older adults who were exposed to caffeine 30 minutes before testing were able to perform on a California Verbal Learning test (CLVT) equally as well in the late afternoon as they did in the morning, whereas participants not exposed to caffeine showed a significant decline [14]. An alternative explanation is discussed by Kemtes and Allen who demonstrated an auditory superiority effect in younger and older populations during exposure to WAIS digit span tests [34].

This phenomenon was observed in our afternoon study where placebo males and females recorded significantly higher scores during auditory tests than visual tests. Several limitations should be noted when considering the study results. Due to time constraints, only two trials were run. Like all drugs administered orally, stomach content can affect absorption levels, which was not considered. Due to time constraints, subjects were only given thirty minutes between administration of their pill and the beginning of their tests, although this timetable falls between the average maximum time and minimum time needed to achieve peak serum caffeine levels, actual levels are unknown due to inability to test blood directly. Due to resource constraints, the school library was used as a testing center, only ten participants could take the test simultaneously and the testing environment was not always quiet. All participants in the study were medical students, although they were all healthy there might be a selection bias skewing the results.

Conclusion

In conclusion, past studies have demonstrated discrepancies among the effects of caffeine. Previous authors attributed these differences to a plethora of variables including, but not limited to, gender, age, fatigue, hormone levels, and time of day, weight, test type, and dosage of caffeine. Our study reveals statistically significant results in the digit span test between males and females as well as the morning and afternoon trials. Much of our data and research suggests that innate differences between test subjects can provide a potential explanation for these findings.

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Medical and Medicinal Journal

The Review about the Kinetics of Monoterpenes Formation in Plants of Sort Artemisia

Essential oils from plants of sort Artemisia are applicable in various branches of perfumery, pharmaceutical and the food-processing industry. Predominant components and their content define practical value of essential oil. Many explorers [1-3] studied the oils composition obtained from plants of sort Artemisia. The basic valuable components of essential oils are various representatives of monoterpenes. The huge actual data about the monoterpenic reproduction during the vegetative period of various ethereous-carrier plants is saved up. But numerous researches, as a rule, confine some selves by the only exposition of exact predominant components within the limits of any taxon-carrier for practical purposes [4-6]. In the present review the author’s kinetic model affixed to the reproduction of monoterpenic hydrocarbons in life cycle of five Crimean plants of sort Artemisia [7-9] and to that from Eastern Siberia (A.glauca) [10] is used. Monoterpenes possess various biological effect: one with an open chain are used in perfumery (citral, geraniol, linalyl ester) and other are used in the food-processing industry as gustatory components. Between cyclical monoterpenes there are widely known medical products: menthol, terpin hydrate (monocyclic), camphor (bicyclic) [11-13]. Composition of monoterpenic components and their content in plants define as the quality and the industrial utility of received essential oil. By the method of hydro distilling from plants of type Artemisia abrotanum, A.annua, A.dracunculus, A.glauca, A.taurica, A.scoparia essential oils had been received, and then they are parted by the chromate-mass spectroscopy on components and are identified.

Then monoterpenic components of essential oils have been distributed according to their ring formation degree. For each degree of ring formation, as shown in the article [14], one calculated the normalized concentrations according to vegetation phases, namely: I – the vegetation beginning, II – budding, III – mass blooming, IV – maturing of seeds. Thus, one obtained the accumulation mass fractions of not exact compounds in studied plants, but the generalized carbocyclic structures, such as: acyclic (X2), monocyclic (Y2) and bicyclic (Z) (Figure 1), which can be extrapolated on all plants and to evolve the general regularities of their accumulation. The finding of singularities in a connatural monoterpenes biosynthesis was made by own kinetic model reduced in the article [15,16]. Data about the qualitative composition and the quantitative maintenance of monoterpene’s components in the essential oils obtained from plants of sort Artemisia balchanorum Krasch was given in the article [15]. Such experimental data about the essential oil’s composition have been compared with the steps of calculated curves so that these curves must be maximum corresponded to the data (Figure 1) according to the known algorithm of the regressive analysis (Fisher’s criterion), reduced in the articles [15,16]. The received graphic solutions of a differential equations system represent those selves a data bank of monoterpenes accumulative dynamics in the plants growing as in the Crimean region and in Eastern Siberia also. Thus, the quality and quantitative content of monoterpene’s hydrocarbons in two sort plants samples of Artemisia balchanorum is detected during all phases of vegetation with the distribution of the components according to the degree of cyclization and with the definition of their group quantity properties and it is placed on the (Figure 1).

Also, we constructed the similar calculated curves for the extract compositions from some other plants samples of vegetable species Artemisia balchanorum according to their ring formation degree during all phase’s vegetation (the phases I-IV), as it is demonstrated on the (Figure 2). The steps of graphic curves (1) (Figures 1 & 2) indicate on the possibility to describe vegetative types by the kinetic methods according to the graphic maps, which demonstrate the efficiency of the investigated monoterpenes groups. There are accurate enough graphic differences between six types of the plants, despite the uniform mechanism of biosynthetic process (Figure 2). The constants of biosynthesis and the factor planes for each vegetative type are presented in the article [15]. For the medical industry it is more perspective to cultivate just the plant A. glauca Pall. ex Willd, which is rich monocyclic terpenes (β -fellandren, γ-terpinen, terpinolene) for use that as medical products [10-13]. There is a wide spectrum of benefit performance biological properties [17], which is corresponded to the components of essential oils from plants A. annua L., such as A. abrotanum L., A. dracunculus L., A. scoparia Waldst. et Kit. Artemisia Taurica Willd. Data of Figures 1 & 2 show a possibility to organize the harvesting of these plants such way that its results will lead to demanded composition of monoterpenes in the essential oils according to the corresponded vegetative phase.

One chooses some perspective sort-samples of these plants, which are used now as the agronomic culture for a pharmacology. For example: the sort Beginner in the type of A.annua L. is rich acyclic components, namely: 1,8-tsineol, Artemisia ketone, Artemisia alcohol having a high quantity of bicyclic camphor ketone; the sort Evksin in the type of A.abrotanum L. contains 1,8-tsineol and α -thujone as principal components of essential oil, is designed to be used not only in a medicine, but also in a perfumery, and also in the food-processing industry; one made two sorts named as WIM and the Crystal in the type of A.dracunculus L., which contain principal components, such as linalyl ester (at one) and neril-acetate (at another); the sort Branching in the type of A.scoparia Waldst.et Kit contains a high quantity of capillene in the essential oil; the sort Alupka in the type of A.taurica Willd contains bicyclic monoterpenes (thujones are the principal components and the camphor is presented also) [18]. The received outcomes about a monoterpenic hydrocarbons reproduction during the life cycle of ethereous-carrier plants will form a data bank of qualitative and quantitative composition of monoterpenic components in the essential oils from five Crimean plants types of sort Artemisia and one of that from Eastern Siberia. It is offered performances of vegetative types using the graphic maps of their efficiency according to degree of ring monoterpenes formation. It is offered to use the considered data bank at a choice of the plants sort-examples to cultivate them for exact industrial needs, to plane the terms of plants harvesting on purpose to extract from them the essential oil possessed by the known properties, to do a forecasting and to study the singularities of plants vegetation in agricultural region and to plane the compositions from the oils for industrial needs [19].

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Top Medical Open Access Journal

Buffalo Milk Adulteration in South Punjab (Pakistan) -A Public Health Hazard

Buffalo milk, one of the substantial known human foods, provides many nutrients such as carbohydrates, fats, proteins, vitamins, minerals, and other important components (Yasmin et al., 2012). Buffalo milk is popular in Pakistan and other south Asian countries. Buffalo milk is unique due to less cholesterol, high fat, content more calories that add to its health benefits. Moreover, it is a good diet for developing healthy bones, maintaining dental health, and preventing cardiovascular disorders. Buffalo milk is thick and creamy in consistency therefore suitable for manufacturing traditional dairy products including yogurt, cottage cheese (paneer) and traditional milk products like ghee and khoa (Haque [1]). According to authors, shorter summers (hence shorter periods of pasture grazing) in the Nordic countries could explain CLA differences between the Nordic and other European countries. However, it was not known if such changes would occur under tropical conditions with the traditional feeding system of Sindh, (Pakistan), where animals are fed on forages throughout the year, except in winter and late spring when shortage of green forage occurs, and ruminants are fed crops by product along with available green fodder. Pakistan ranks as the 5th largest producer of milk in the world with over 26 million buffaloes, 56 million sheep, 24 million of each cattle and goats. Buffaloes are the major milk producing animal, accounting for about 75% of all milk produced. They are concentrated in irrigation areas and long rivers, as are the human population.

Goat and ewe milk are not proceeded for commercial purposes in Sindh (Pakistan), and these animals are mostly reared for meat purpose, while milking is a secondary function, which provide extra source of income to poor farmers (Sarwar et al., 2002). However, data on milk fatty acid composition from ruminant species from Pakistan is scarce. Milk adulteration became a global concern after breakthrough of melamine adulteration in Chinese infant milk formula. Worldwide, it is a terrible situation that milk is being very easily adulterated and the situation is significantly worse in underdeveloped countries due to lack of adequate monitoring and absence of proper law enforcement (Xin [2]). Dairy Milk adulteration is a socioeconomic issue in developing countries. The act of adding adulterants makes it unfit for human consumption and dairy products fail to meet the legal standards. It brings not only brings unfavorable consequences in terms of major economic losses for the processing industry, but also a major health hazards for the consumers from infants to adults worldwide. The consumption of adulterated milk causes several critical health disorders including dysentery, colon ulcers, nephrosis, disturbance of cardiovascular system (Hanford [3]). In Pakistan limited studies are available on level of dairy milk adulteration, a major public health hazard. Limited studies have been conducted on buffalo milk composition variation in different seasons but none in south Punjab where summer season lasts for long duration and winter lasts for short period.

The first objective of the study was to estimate variation in composition of buffalo milk. The second objective was to know about adulterant types used in buffalo milk supplied for human consumption in South Punjab region of the country.

Methodology

A total of 100 milk samples were collected by convenience sampling method from various cities in southern Punjab. Milk samples were collected in two seasons. Winter(n=50) sampling was conducted from December to March and summer(n=50) sampling was conducted from April to June from same cities. Milk samples were collected from dairy farms(n=40), local vendors(n=30), and commercial chillers(n=30). The raw milk samples were collected from homogenized milk into sterilized Falcon tubes to avoid any type of contamination. All milk samples were labelled and placed in ice filled cooler box. The samples were transported to the Milk Testing Laboratory Punjab Food Authority, Multan and preserved at 4-8°C until analysis. For estimation of milk fat, protein, lactose, and total solids Lactoscan milk analyzer was used (Miltonic Ltd.series7035, Bulgaria). For detection of adulterants (starch, urea, hypochlorite, pulverized soap, formalin, sugar, skim milk, boric acid and detergent) user friendly Latte Adulterazione kit was used according to the manufacturer’s protocol. The data generated was entered in software SPSS and independent t test was applied to find significant difference in milk composition.

Results

In the present study, change in milk composition during extreme weather conditions of South Punjab region were investigated. Data shows that fat content of milk samples was significantly (P<0.05) different in winter and summer seasons. Milk protein content was also significantly(P<0.05) different in winter and summer season. Milk lactose content was non significantly (P>0.05) different in winter and summer seasons. Milk total solids content showed significant (P<0.05) difference in winter and summer season as shown in Table 1. In the present study data from dairy farms showed no milk adulteration in winter and summer season in South Punjab region. Milk samples acquired from local vendors showed use of skim milk followed by detergent as adulterants were high both in winter (52.5%;33.33%) and summer (72%;31.33%) seasons. Milk samples acquired from commercial chillers showed use of skim milk followed by starch as adulterants were high both in winter (60%; 33.33%) and summer season (60%; 33.33%). The use of urea as milk adulterant was found in summer (28.3%) in buffalo milk samples as shown in Table 2.

Note: P≤0.05 (*shows that value is significant)

Discussion

Go to

Out of total dairy milk produced in Pakistan, buffalo contributes about 68 %, followed by other dairy animals. Due to the high fat contents of buffalo milk, it is the most preferred by people among other dairy products. In the present study buffalo milk samples showed difference in fat, protein, and total solids contents in winter and summer seasons. Saadi, et al. (2019) finding is in line with our study and show significant difference (p<0.001) with highest percentage of fat 4.48 % in milk in winter while values decrease to 2.95% in summer. The decrease in fat in summer may be due to the length of light compared to dark. The higher the ratio between light and dark the lower the proportion of fat due to increased prolactin secretion whose concentration in plasma is higher in summer than in winter (Ozrenck [4]). As for the effect of nutrition the basis in the composition of fat depended on the composition of acetate in the rumen. Food such as grain feeding that reduces acetate production will also reduce fat concentration in milk produced during the summer (Vildirim and Cimen, 2009). Percentage of protein was highest in milk produced from cows, which fed on concentrated feeds in the winter and amounted to 3.46% and decreased to 2.93% for the milk of cows that fed on herbs in the summer, and attributed the reason for high protein in the winter to the diet content high in protein while in the summer the nutrition was low in protein content (Colombari [5]). The low percentage of fiber in cattle (concentrated feed) resulted in increased protein content in milk produced. On the contrary, when they fed on green the protein decreased (Petitclerc, et al. 2000). Significant differences were found in the percentage of total solids, as the proportion of fat increased the proportion of total solids (Pavel [6]). The freezing point and lactose sugar showed no significant difference. Difference in milk composition in both seasons might be due to change in feed and water consumption by buffaloes (Afzal [7,8]). In the present study buffalo milk adulteration was observed in both seasons. Skim milk powder was the most used adulterant for thickening of milk in the study area as reported in literature (Azad [9]). Urea added in buffalo milk to increase non protein nitrogen content. Urea adulteration has carcinogenic effects on human health. Starch is used as a thickening agent in milk. Starch adulteration can cause diarrhea and its accumulation in the body might be fatal for diabetic patients (Yadav [10]). Detergents are added to emulsify and dissolve the oil in water to give frothy solution, a desired characteristic of milk. Consumption of adulterated milk leads to kidney failure, gastritis, and intestinal inflammation (Rennie [11-14]).

Conclusion

People living in developing countries like Pakistan, which is already far behind in delivering health services, are deprived of pure healthy buffalo milk. The adulterated milk is of low quality and responsible for introducing hazardous substances leading to serious health hazards in consumers. Therefore, it is need of the hour to devise an efficient and reliable quality control system that will regularly monitor the activities of malpractices in the dairy industry. It is the responsibility of the government to formulate an effective strategy to ensure the access of fresh and quality raw milk to people so that infants and adults both can enjoy a healthy life by having pure natural milk. General public awareness, effective monitoring measures and regulatory system for quality control of milk and dairy products can play a crucial role in minimizing milk adulteration.

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American Medical Journal

The Impact of Gut Microbiota on Long COVID: Insights and Challenges

Methods

Results and Discussion

Conclusion

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