Density functional theory (DFT), molecular docking, and xanthine oxidase inhibitory studies of dinaphthodiospyrol S from Diospyros kaki L.

In this work, we investigated Diospyros kaki extract and an isolated compound for their potential as xanthine oxidase (XO) inhibitors, a target enzyme involved in inflammatory disorders. The prepared extract was subjected to column chromatography, and dinaphthodiospyrol S was isolated. Then XO inhibitory properties were assessed using a spectrophotometry microplate reader. DMSO was taken as a negative control, and allopurinol was used as a standard drug. The molecular docking study of the isolated compound to the XO active site was performed, followed by visualization and protein-ligand interaction. The defatted chloroform extract showed the highest inhibitory effect, followed by the chloroform extract and the isolated compound. The isolated compound exhibited significant inhibitory activity against XO with an IC50 value of 1.09 µM. Molecular docking studies showed that the compound strongly interacts with XO, forming hydrogen bond interactions with Arg149 and Cys113 and H-pi interactions with Cys116 and Leu147. The binding score of -7.678 kcal/mol further supported the potential of the isolated compound as an XO inhibitor. The quantum chemical procedures were used to study the electronic behavior of dinaphthodiospyrol S isolated from D. kaki. Frontier molecular orbital (FMO) analysis was performed to understand the distribution of electronic density, highest occupied molecular orbital HOMO, lowest unoccupied molecular orbital LUMO, and energy gaps. The values of HOMO, LUMO, and energy gap were found to be -6.39, -3.51 and 2.88 eV respectively. The FMO results indicated the intramolecular charge transfer. Moreover, reactivity descriptors were also determined to confirm the stability of the compound. The molecular electrostatic potential (MEP) investigation was done to analyze the electrophilic and nucleophilic sites within a molecule. The oxygen atoms in the compound exhibited negative potential, indicating that they are favorable sites for electrophilic attacks. The results indicate its potential as a therapeutic agent for related disorders. Further studies are needed to investigate this compound's in vivo efficacy and safety as a potential drug candidate.

Edible mushrooms as potential functional foods in amelioration of hypertension.

Edible mushrooms are popular functional foods attributed to their rich nutritional bioactive constituent profile influencing cardiovascular function. Edible mushrooms are omnipresent in various prescribed Dietary Approaches to Stop Hypertension, Mediterranean diet, and fortified meal plans as they are rich in amino acids, dietary fiber, proteins, sterols, vitamins, and minerals. However, without an understanding of the influence of mushroom bioactive constituents, mechanism of action on heart and allergenicity, it is difficult to fully comprehend the role of mushrooms as dietary interventions in alleviating hypertension and other cardiovascular malfunctions. To accomplish this endeavor, we chose to review edible mushrooms and their bioactive constituents in ameliorating hypertension. Hypertension and cardiovascular diseases are interrelated and if the former is managed by dietary changes, it is postulated that overall heart health could also be improved. With a concise note on different edible varieties of mushrooms, a particular focus is presented on the antihypertensive potential of mushroom bioactive constituents, mode of action, absorption kinetics and bioavailability. Ergosterol, lovastatin, cordycepin, tocopherols, chitosan, ergothioneine, γ-aminobutyric acid, quercetin, and eritadenine are described as essential bioactives with hypotensive effects. Finally, safety concerns on allergens and limitations of consuming edible mushrooms with special reference to chemical toxins and their postulated metabolites are highlighted. It is opined that the present review will redirect toxicologists to further investigate mushroom bioactives and allergens, thereby influencing dietary interventions for heart health.

Recent advances in the therapeutic application of short-chain fatty acids (SCFAs): An updated review.

Over the past decade, the gut microbiota has emerged as an important frontier in understanding the human body's homeostasis and the development of diseases. Gut flora in human beings regulates various metabolic functionalities, including enzymes, amino acid synthesis, bio-transformation of bile acid, fermentation of non-digestible carbohydrates (NDCs), generation of indoles and polyamines (PAs), and production of short-chain fatty acids (SCFAs). Among all the metabolites produced by gut microbiota, SCFAs, the final product of fermentation of dietary fibers by gut microbiota, receive lots of attention from scientists due to their pharmacological and physiological characteristics. However, the molecular mechanisms underlying the role of SCFAs in the interaction between diet, gut microbiota, and host energy metabolism is still needed in-depth research. This review highlights the recent biotechnological advances in applying SCFAs as important metabolites to treat various diseases and maintain colonic health.

Maternal and peri-natal outcomes of dengue fever with special emphasis on vertical transmission.

OBJECTIVE: To investigate the maternal and peri-natal outcomes of dengue infection and frequency of dengue immunoglobulin G positivity in pregnant women along with pregnancy outcomes. Method: The observational two-phase study was conducted from 2012 to 2015 in Lahore, Pakistan. In phase 1, pregnant women who had acute dengue fever were prospectively and retrospectively enrolled from 4 tertiary care hospitals. Demographic data, clinical/laboratory parameters and maternal/foetal outcomes were recorded for each subject. In the second phase, normal pregnant women with no current or past history of dengue fever were enrolled from Gynaecology Department of Shaikh Zayed Hospital, Lahore, to determine the frequency of dengue immunoglobulin G positivity and pregnancy outcome. Data was analysed using SPSS 20. RESULTS: Of the 12 subjects in phase 1, 10(83.3%) were prospective cases and 2(16.6%) were retrospective. The overall mean age was 26.3±5.4 years, 7(58.3%) were in 2nd and 5(41.6%) were in 3rd trimester of pregnancy. Among the 4(33.3%) women at gestational age ≥33 weeks, 3(75%) had poor foetal outcome. In the second phase, there were 127 women with a mean age of 27.4±4.3 years. Among them, 38(29.9%) women were in the 2nd and 89(70.1%) were in the 3rd trimester. Dengue immunoglobulin G positivity was found in 65(51.2%) cases, but poor maternal or foetal outcome was not found in such women. CONCLUSIONS: Acute dengue fever at late term resulted in adverse foetal outcomes. Frequency of dengue immunoglobulin G positivity was high among pregnant women, but it was not associated with adverse outcomes.

Pregnancy outcomes with confirmed SARS-CoV-2 infection during first wave: A review.

Maternal and foetal care has become an important concern in the wake of enormous global spread of coronavirus disease-2019 (COVID-19), but there is scarcity of information about maternal and perinatal outcomes. The current review was conducted from March to July 2020. Appropriate and related databases were searched electronically by using terms, like "COVID-19 and pregnancy", "pregnancy outcomes of COVID-19". Pooled analysis of the reviewed studies showed that of the 164 newborns, vertical transmission was noted in 7(2.95%). The most common element 140(84.98%) was caesarean section deliveries. COVID-19 pneumonia developed in almost 54(30.90%) of 175 women. The most common symptom of COVID-19 among women was fever 88(50.77%). Adverse maternal and foetal outcomes were found to be associated with COVID-19 in the form of severe illness, increased rates of caesarean section deliveries and worse birth outcomes. Yet, vertical transmission of COVID-19 infection remains debatable.

Dietary patterns of Pakistani adults and their associations with sociodemographic characteristics-a community based study.

OBJECTIVE: To identify the dietary patterns and understand their association with sociodemographic characteristics among adults. METHODS: The community-based cross-sectional study was conducted in Islamabad, Lahore, Karachi, Peshawar and Quetta cities of Pakistan from March to November 2018, after approval from the National Bioethics Committee, Islamabad, and comprised adults of either gender. Data was collected using the food frequency questionnaire, and dietary patterns were identified using factor analysis. Multivariate regression analysis was used to assess the association of socio-demographic determinants with dietary patterns. Data was analysed using SPSS 21. Parallel Analysis criterion (Eigen values) was determined along with Monte Carlo simulation. RESULTS: Of the 448 subjects, 206(46%) were males and 242(54%) were females. The largest age group was 36-55 years 199(47.4%). Six dietary patterns were identified: "Vegetables", "Fruits", "Mixed Junk and Processed food", "Dairy and Fast food", "Discretionary" and "Fish". Regression analysis showed that those aged 36-55 years had higher scores for vegetables, fruit and fish pattern (p<0.05). Females scored more for vegetables, fish and fruits, and a significantly low score for discretionary diet pattern (p<0.05). Participants with high education level and socioeconomic status had raised scores for discretionary diet items (p<0.05). CONCLUSIONS: Six distinct dietary patterns among Pakistani adults were identified, showing significant association with sociodemographic characteristics.

Junk food consumption in relation to menstrual abnormalities among adolescent girls: A comparative cross sectional study.

Objectives: The study was aimed at evaluating the association between junk food consumption and BMI of adolescent girls along with the menstrual abnormalities and to compare it with controls. Methods: A cross-sectional study was conducted among 200 girls between 13 - 19 years of age at Bahria International Hospital, Lahore based on self-administered questionnaire from July 2021 to September 2021. The total subjects were divided in two groups Viz; Group-A which comprised of 100 girls with menstrual abnormalities and Group-B included 100 girls without menstrual problem (control group). The data recorded on the questionnaire about the demographic profile, anthropometric measurements, menstrual cycle characteristics, and dietary habits was subjected to statistical analysis using SPSS version 20 and Chi-Square was used to test quantitative significance between the two groups. Results: The mean age of participants was 17.02±1.76 years. It was observed that 40% girls had irregular menstrual cycle, 56% girls were suffering from dysmenorrhea and almost all girls of Group-I were suffering from premenstrual dysfunctions. The current study found a non-significant difference between two groups with regard to body mass index (P≥0.05). Significant difference was observed between two groups (P ≤ 0.05) as junk food consumption was high in Group-A as compared to Group-B. However, no significant difference was found between Group-A and B in relation to the consumption of salty snacks and frozen meat items (P≥0.05). Conclusion: The results suggested that junk food consumption affects menstrual cycle negatively however more studies are needed to confirm the association of BMI, consumption of salty snacks and frozen meat items with menstrual abnormalities.

Amino acid starvation sensing dampens IL-1ß production by activating riboclustering and autophagy.

Activation of the amino acid starvation response (AAR) increases lifespan and acute stress resistance as well as regulates inflammation. However, the underlying mechanisms remain unclear. Here, we show that activation of AAR pharmacologically by Halofuginone (HF) significantly inhibits production of the proinflammatory cytokine interleukin 1ß (IL-1ß) and provides protection from intestinal inflammation in mice. HF inhibits IL-1ß through general control nonderepressible 2 kinase (GCN2)-dependent activation of the cytoprotective integrated stress response (ISR) pathway, resulting in rerouting of IL-1ß mRNA from translationally active polysomes to inactive ribocluster complexes-such as stress granules (SGs)-via recruitment of RNA-binding proteins (RBPs) T cell-restricted intracellular antigen-1(TIA-1)/TIA-1-related (TIAR), which are further cleared through induction of autophagy. GCN2 ablation resulted in reduced autophagy and SG formation, which is inversely correlated with IL-1ß production. Furthermore, HF diminishes inflammasome activation through suppression of reactive oxygen species (ROS) production. Our study unveils a novel mechanism by which IL-1ß is regulated by AAR and further suggests that administration of HF might offer an effective therapeutic intervention against inflammatory diseases.

Vertical Transmission of SARS-CoV-2 from COVID-19 Infected Pregnant Women: A Review on Intrauterine Transmission.

BACKGROUND: Fetal safety is a major concern with the global spread of COVID-19, but there is scarce information regarding vertical transmission and how it affects the fetus. OBJECTIVE: To assess and summarize the currently available evidence on vertical transmission (probable/confirmed) of SARS-CoV-2 along with fetal outcomes. METHODS: The current review was carried out from March to October 2020. Relevant databases were searched electronically. Pertinent articles were selected according to eligibility criteria and information was compiled. RESULTS: In 16 selected articles there were total 498 COVID-19 infected pregnant women ranging in age between 15 and 45 years. Gestational age at the onset of COVID-19 symptoms ranged from 25-41 weeks. Vertical transmission (probable and confirmed) rate from series was 4.883% (23/471). Of 17 affected newborns (information available), 08 required NICU admission, 04 developed pneumonia and 04 required mechanical ventilation. CONCLUSION: There is probable intrauterine transmission of SARS-CoV-2. Few adverse fetal outcomes are associated with COVID-19.

Routes and barriers associated with protein and peptide drug delivery system.

Proteins and peptide drugs have a great therapeutic potential and their usage in the treatment of various severe diseases has revolutionised the fields of pharmaceuticals and biotechnology. For successful therapeutic effects, various efforts have been made for effective delivery of proteins/peptide drugs through various routes of administrations. Parenteral and non-parenteral drug deliveries are regarded as significant routes of drug absorption. In addition to intravenous, subcutaneous and intramuscular routes, the oral route is more effective for protein and peptides therapeutics. However, there is a need to improve non-parenteral drug delivery systems (DDS) to increase drug absorption in a more effective way. The present narrative review was planned to describe routes and barriers for protein/peptide drugs and how to improve drug delivery systems in an effective way. For this purpose, numerous research articles were searched from year 2000-2021 using search engines like PubMed, Google Scholar, Medline and ISI Web of Knowledge, and Bioline International while applying different keywords such as 'protein and peptide drugs', 'drug delivery systems', 'parenteral and non-parenteral routes of drug delivery' and 'physicochemical barriers'. It was concluded that the success of the therapeutics is strongly influenced by the differential delivery of targeted antigen, the choice of targeting protein or peptide, and drug-release characteristics of the linker used. Furthermore, there should be an improvement in non-parenteral DDSs so that the drugs might be administered in an appropriate manner.

Activation of integrated stress response pathway regulates IL-1ß production through posttranscriptional and translational reprogramming in macrophages.

Immune cells sense and programme its cellular machinery appropriately to the environmental changes through the activation of cytoprotective adaptive pathway so-called the "integrated stress response (ISR)". However, the mechanisms implicated in ISR-induced protective responses are poorly understood. Here, we show that ISR activation by arsenite (Ar) results in suppression of IL-1ß production in macrophages and inhibition of DSS-induced colitis in a murine model through a novel posttranscriptional and translation regulatory (PTR) mechanism. Ar triggers PTR events through eIF2α-phosphorylation, which results in the attenuation of active polysome formation leading to the accumulation of translationally stalled IL-1ß mRNAs. Translationally stalled IL-1ß mRNAs recruit RNA-binding proteins (TIA-1/TIAR), resulting in the formation of RBP-RNA complexes known as stress granules (SGs). The SGs bound IL-1ß mRNAs might undergo degradation through induction of autophagy. Also, we show that Ar posttranslationally impairs processing and secretion of IL-1ß by diminishing inflammasome activation. Altogether, this study unveils a novel mechanism of IL-1ß regulation and further suggests that pharmacological activation of cytoprotective ISR pathway might provide an effective therapeutic intervention against inflammatory diseases.

Transcriptome meta-analysis identifies immune signature comprising of RNA binding proteins in ulcerative colitis patients.

Ulcerative colitis (UC) is a persistent inflammatory illness, which is clinically categorised as Inflammatory bowel disease (IBD), affecting millions of people worldwide. The precise cause behind the pathology of the disease remains unknown. However, the involvement of multiple factors including genetic predisposition, immunological deregulations, microbiota imbalance, and environmental triggers has been suggested. Amongst all these factors, the over-active immunological response reported in UC patients seems to be a promising target for therapy. Moreover, identification of gene signatures associated with disease onset and progression would help in better understanding of the molecular mechanisms involved in the disease pathogenesis. Here, we have conducted meta-analysis of gene expression profiles of UC patient microarray datasets accessible in public databases and further validated the in-silico findings in UC patients' blood samples. Our study reveals that UC pathogenesis perturbs expression of several inflammatory genes. In addition, we report a novel gene signature comprising of TIA1 (T cell restricted intracellular antigen) and TIAR (TIA1 related protein; also known as TIAL1), which were found to be significantly downregulated in UC patients. TIA1 and TIAR are RNA-binding proteins (RBPs), which function as a translational represser by binding to ARE sequences in the 3' UTR of mRNAs encoding inflammatory mediators including cytokines. Our findings demonstrate that deletion of TIAR using gene specific siRNAs in-vitro results in enhanced production of inflammatory cytokine IL-1ß. In conclusion, the findings of this study reveal that down regulation of TIA1/TIAR genes could be responsible for UC associated inflammation. This study highlights the usefulness of the meta-analysis approach in the identification of unique gene signatures that might deliver mechanistic insights into UC pathogenesis and possibly assist in discovery of prognostic markers and therapeutic interventions.

Comparison of Tobacco Consumption among Adults in SAARC Countries (Pakistan, India and Bangladesh).

OBJECTIVE: To compare the findings of Global adult tobacco surveys (GATS) conducted in SAARC Countries. METHODS: The national representative data from Pakistan, India, and Bangladesh was used as GATS was done in these three countries of SAARC. The key variables were selected where information was consistently taken like current smokers, smokeless tobacco users, exposure to second-hand smoke, advise by health care provider to quit, monthly expenditure and noticing of health warning label and other information related to daily tobacco users (smoke, smokeless) mean age of initiation and mean number of cigarettes consumed per day were also collected. Chi-square test was applied and p-value was considered significant at <0.05. RESULTS: Overall tobacco consumption was significantly high (43.3%) in Bangladesh (Pakistan 19.1% and India 34.6%). Similarly, current smokers were significantly more in Bangladesh 23% (India 14% and Pakistan 12%) and smokeless tobacco was significantly more in Bangladesh 27.2% (India 25.9% and Pakistan 7.7%). Exposure to second-hand smoke at work was 69.1% in Pakistan (63% in Bangladesh and 29.9% in India). Monthly expenditure on cigarettes was also high in Pakistan 7.51 USD (India 6.26 USD and Bangladesh 4.57USD). Mean age of initiation was 18.8 years in Bangladesh, (17.8 India and 18.7 Pakistan). Mean number of cigarettes consumed was significantly high in Pakistan i.e. 13.6 (6.2 in India and 5.1 Bangladesh). CONCLUSIONS: All the three South Asian countries have a high prevalence of tobacco consumption. Tobacco use was high in Bangladesh but smokeless tobacco among males was high in India and mean number of cigarettes daily was high in Pakistan.

Prevalence of Smokeless Tobacco Use in Pakistan: Insight from the Global Adult Tobacco Survey Pakistan (GATS Pakistan-2014).

OBJECTIVE: Prevalence of tobacco consumption is increasing including both smoked and smokeless tobacco. The objective of this paper is to highlight the prevalence of smokeless tobacco use, its demographic trends as well as socioeconomic determinants based on Global Adult Tobacco Survey 2014 (GATS) -Pakistan. METHODS: Secondary analysis of GATS data was done which was a nationally representative survey, conducted among non-institutionalized males and females aged 15 years and above. The survey included a total of 9,856 households which were selected using multistage stratified cluster sampling technique. RESULTS: Secondary analysis of GATS data revealed that 8.6% of the Pakistani population was current users of smokeless tobacco; among them, 8.0% were daily users and 0.6% were less than daily users. Prevalence of SLT use was higher in males (13.7%) compared to females (3.9%) (p = 0.000). Mean age of male users was 39.96±14.43 years and of females was 47.71±16.21 (p=0.000). Males (3.959 95% CI = 3.291-4.764) and respondents with primary or less education (2.216 95%CI = 1.841-2.668) were at higher odds of SLT use prevalence. Among daily users, Naswar was the preferred (71.8%) SLT product in males, compared to females (p = 0.000). Whereas, in females, use of Paan with tobacco (26.4%) (P = 0.000) and Naas (20.9%) (P = 0.000) was more common compared to males. Among SLT users, 71.1% participants used to spend 1-100 Rs (0.0095-0.95USD) per week on purchase of SLT products. However 28.9% were spending more than one dollar on the purchase of SLT products including 19.1% who were spending Rs 101-200 (0.96-1.9USD) followed by 7.2% spending Rs 201-300 (1.91-2.85 USD) and 2.6% spending Rs. >300 (>2.85 USD). CONCLUSIONS: Males and less educated individuals should be targeted with behavioral interventions for control of SLT use. Males started SLT use at an earlier age compared to females which can result in premature morbidity and mortality in males. SLT products are cheaper compared to cigarettes, therefore, taxes need to be levied on SLT products.

Exposure of the Adult Pakistani Population to Second-Hand Smoke - An insight from Global Adult Tobacco Survey of Pakistan 2014.

OBJECTIVE: About one-third of the world population is exposed to second hand smoke (SHS) with estimated 600,000 annual deaths. The objective of this article is to determine the exposure of adult Pakistani population to SHS and its association with different demographic variables based on data of Global Adult Tobacco Survey (GATS) conducted in 2014 in Pakistan. METHODS: Secondary analysis of Global Adult Tobacco Survey (GATS) data was done which is a global standard used for systematically monitoring use of both smoked and smokeless tobacco products. GATS Pakistan was a nationally representative survey, conducted in all four provinces among adult males and females aged15 years and above. Using multistage stratified cluster sampling technique a total of 9,856 households were selected and finally 7,831 individuals were enrolled. RESULTS: At homes around 43.3% individuals were exposed to SHS. Univariate analysis revealed that overall males (Odds Ratio, OR: 1.17 CI: 1.04-1.30, p=0.006) and less educated (OR: 1.30, CI: 1.16-1.46, p=0.000) group were at higher odds of being exposed to SHS at homes. Over all exposure to second hand smoke at home was also significantly high among urban residents (p = 0.000). Among nonsmokers age group 15 -35 years (odds ratio, 1.24 CI:1.09-1.04, P=0.01) and less educated group (OR 1.24, CI: 1.09-1.40, p=0.001) were at higher odds of being exposed to SHS at home At indoor workplaces, overall 69.1% and among non-smokers, 65.3% individuals were exposed to SHS. Univariate analysis has shown that less educated group (OR: 1.525, CI: 1.012-2.298, p=0.043) was at higher odds of being exposed to SHS at indoor workplaces. Among various public places exposure of participants to SHS was highest (86%) at restaurants, followed by public transport (74%), marriage halls (65%), universities (46%), health care facilities (35%) and least at schools (20%).. CONCLUSIONS: Less educated males were more exposed to second-hand smoke both at work places and public places; therefore to start with interventions, work places and public places should be a priority as at home results could be confounded by self-smoking as well.

Determinants of Smoked Tobacco Consumption among Pakistani Adults: An Analysis from GATS Pakistan 2014.

OBJECTIVE: An in-depth analysis of GATS 2014 is performed to study the various demographic determinants of smoked tobacco consumption among Pakistani adults. METHODS: The Global Adult Tobacco Survey (GATS) is a standardized surveillance mechanism regarding monitoring of tobacco consumption globally, that uses three stage cluster random sampling technique to have a representative data. We performed the complex data set analysis using SPSS V 15. The associations were determined by carrying out univariate analysis, and determining chi squares for categorical variables. RESULTS: Overall current smoking prevalence was found to be 12.4%. The univariate analysis revealed male gender (OR: 14.1 CI= 11.2-17.7, p value 0.000), little or no education (OR: 1.4 CI=1.2-1.6, p-value 0.000), living in urban areas (OR: 0.7 CI= 0.6-0.8, p-value 0.000) and avoiding smoking attempts in young age till 25 years (OR: 0.2 CI= 0.1-0.2, p value 0.000) were determinants. Assosiation of demographic determinants with cessation behaviour showed that the health concern was the primary reason to ever stop smoking among both urban rural dwellers, educated and uneducated and respondents aged 25 years and above. Educational status, age of respondent and residence had almost no significant effect on cessation behaviour of smoked tobacco users. CONCLUSIONS: Males having low education and living in rural area puts an adult Pakistani at high risk of becoming the user of smoked tobacco. Health education involving primary health care providers particularly focusing on rural areas will reduce the prevalence of smoking.

Vitex negundo induces an anticonvulsant effect by inhibiting voltage gated sodium channels in murine Neuro 2A cell line.

Vitex negundo (Vn) extract is famous for the treatment of neurological diseases such as migraine and epilepsy. These neurological diseases have been associated with abnormally increased influx of sodium ions into the neurons. Drugs that inhibit voltage gated sodium channels can be used as potent anti-epileptics. Till now, the effects of Vn on sodium channels have not been investigated. Therefore, we have investigated the effects of methalonic fraction of Vn extract in Murine Neuro 2A cell line. Cells were cultured in a defined medium with or without the Vn extract (100 µg/ml). Sodium currents were recorded using whole-cell patch clamp method. The data show that methanolic extract of Vn inhibited sodium currents in a dose dependent manner (IC50 =161µg/ml). Vn (100 µg/ml) shifted the steady-state inactivation curve to the left or towards the hyper polarization state. However, Vn did not show any effects on outward rectifying potassium currents. Moreover, Vn (100 µg/ml) significantly reduced the sustained repetitive (48±4.8%, P<0.01) firing from neonatal hippocampal neurons at 12 DIV. Hence, our data suggested that inhibition of sodium channels by Vn may exert pharmacological effects in reducing pain and convulsions.

Development of a high sensitivity C-reactive protein immunoassay and comparison with a commercial kit.

C-reactive protein (CRP) is intricately sensitive marker of inflammation, infection, and tissue damage. Role in the prognosis of heart diseases has been recently discovered. This study aimed to develop a cost-effective and high-sensitivity CRP immunoassay for use in cardiac risk assessment. Assay was optimized for coating, blocking of capturing antibody, dilution, and reaction time of the conjugate and sample volume. For normal reference range, CRP was determined in serum samples from apparently healthy volunteers. For clinical validation, CRP was determined in samples of acute coronary syndrome patients by in-house and commercial assays. The lower detection limit of in-house assay was 0.16 µg/L. Intra and inter assay imprecision was 4.39%, 4.6% and 8.6%, 9.3%, respectively. The correlation between the CRP levels by the two assays was r = 0.861. Sensitivity, specificity, predictive value for a positive test, and a negative test of in-house assay was 95.3%, 92.8%, 95.3%, and 92.8%, respectively. At lower-end CRP levels of both kits correlated very well but showed variation at upper end. In-house assay showed high sensitivity and reliability at lower end and it is hoped that will help to evaluate cardiac risk assessment (after improvement at upper end) in clinically poor settings.

A Comparative Study on Hemp (Cannabis sativa) Essential Oil Extraction Using Traditional and Advanced Techniques.

A comparative study of Cannabis sativa (Hemp) essential constituents obtained by using Supercritical Fluid Extraction (SCFE), Steam Distillation (SD) and Hydrodistillation (HD) is presented here. The optimized extraction temperatures were 130，110 and 50 ℃ for hydrodistillation, steam distillation and supercritical fluid extraction respectively. The essential oil of C. sativa was analyzed by using Gas chromatography mass spectrometry (GC-MS). A total of 33, 30 and 31 components have been identified in HD, SD and SCFE respectively. Yield of essential oil using SCFE (0.039%) was more than HD (0.025%) and SD (0.035%) extraction respectively. The main component of sesquiterpenes obtained by hydrodistillation at 130 ℃ with their percentages included caryophyllene (40.58%), trans-α-bergamotene (5.41%), humulene (10.97%), cis-ß-farnesene (8.53%) and monoterpenes included α-pinene (2.13%), d-limonene (6.46%), p-cymol (0.65%) and cineole (2.58%) respectively. The main component of sesquiterpenes obtained by SD steam distillation at 110 ℃ including caryophyllene (38.60%) trans-α-bergamotene (4.22%), humulene (10.26%), cis-ß-farnesene (6.67%) and monoterpenes included α-pinene (3.21%), d-limonene (7.07%), p-cymol (2.59%) and cineole (3.88%) whereas the more percentages of major components were obtained by SCFE at 50 ℃ included caryophyllene (44.31%), trans-α-bergamotene (6.79%), humulene (11.97%) cis-ß-farnesene (9.71%) and monoterpenes included α-pinene (0.45%), d-limonene (2.13%) p-cymol (0.19%) and cineole (1.38 %) respectively. We found yield/efficiency, chemical composition, quality of the essential oils by supercritical fluid extraction superior in terms of modern, green, saving energy and a rapid approach as compared to traditional techniques.

Chest X-ray: an unfair screening tool.

BACKGROUND: Chest radiography is considered as the sole screening tool for chest pathologies as prerequisite for overseas employment during their medical examinations. The aim was to evaluate the role of CXR as a screening tool in the diagnosis of different lung pathologies in general and TB in particular. METHODS: In this descriptive case series, reports of 63,648 Pakistani candidates, who took chest X-rays during medical screening for employment abroad under the rules of Gulf Coordination Council (GCC), were analysed. The reports were collected over a three year period (2008-2011). All the candidates were apparently healthy males of ages ranging between 18-50 years with average age of 27.4 years. RESULTS: Out of 63,648 candidates examined over a three year period, 1368 (2.15%) were declared unfit on the basis of CXR. There were 359 cases of calcified granulomas, 318 of costophrenic angle obliteration, 174 of apical scaring; 144 of pulmonary scars, 123 of fibrocalcific lesion, 82 of soft shadows in lung, 51 of nodular lung shadows, 42 of lymphadenopathy, 30 of pleural effusion, 26 of interstitial lung shadow and 19 of calcification. A closer scrutiny of these cases revealed that over 90% of the CXRs of the candidates declared as having lung pathologies displayed scars rather than active disease. CONCLUSION: A normal CXR by itself, neither categorically rules out problems in the chest nor does any finding in it confirm some disease. Unfitness on the basis of CXR alone, unless substantiated by further investigation, is questionable to say the least.