Oxidative stress in polycystic ovary syndrome: a case-control study.

OBJECTIVE: To compare the serum levels of biochemical and oxidative stress markers i.e., malondialdehyde (MDA) and paraoxonase-1(PON1) in polycystic ovary syndrome (PCOS) patients and healthy female individuals of reproductive age group (18-40 years). METHODS: This case-control study was conducted in Dow University of Health Sciences (DUHS), Karachi from June 2019 to October 2020. Seventy Subjects including 35 PCOS patients that have primary subfertility problem (cases) and 35 healthy and fertile females (controls) were recruited. Serum samples were collected for analysis of insulin, sex hormone-binding globulin, testosterone, fasting blood glucose and lipid profile. PON 1 and MDA levels were estimated by ELISA. Comparison between the two groups was done using independent t-test. RESULTS: The patients had significantly increased mean body mass index (28.5+4.6 kg/m2 vs 25.7+4.5 kg/m2, p=0.014), systolic (129.6±13.9 mm of Hg vs 113±7.7 mm of Hg, p<0.001) and diastolic (78.7±8.8 mm of Hg vs 74.6±6.7 mm of Hg, p=0.032) blood pressures compared to controls. The high-density lipoprotein cholesterol levels were significantly lower in PCOS (42.2±8.6mg/dl) than controls (48.8±11.8mg/dl, p=0.009, p=0.009). Serum insulin (14.3±5.8 uIU/mL) vs (10.0±5.2 uIU/mL), p=0.002 and testosterone levels (1.3±0.9 nmol/L) vs (0.82±0.3 nmol/L), p<0.001 were significantly higher whereas sex hormone binding globulin (SHBG) levels (35.2±19.7nmol/L vs 58.8±31.0 nmol/L) were significantly lower in patients than healthy controls (p<0.001). Both oxidative stress markers, paraoxonase 1 (7.7±2.4 vs 6.4±2.6 µg/mL, p=0.04) and malondialdehyde (2.5±1.0 vs 1.9+0.51µg/mL, p=0.034) levels were significantly elevated in PCOS patients than controls. No significant correlation was found between dietary habits and life style between cases and controls. CONCLUSIONS: The study reported significantly elevated levels of oxidative stress markers in PCOS patients.

Serum Leptin levels in newly diagnosed Epileptic Patients.

Objectives: To compare leptin levels in newly-diagnosed treatment-naive epileptic patients and healthy controls. METHODS: This case-control study was conducted from January 10 to September 15, 2022, at the Dow University of Health Sciences, Karachi, and comprised newly-diagnosed epileptic patients age 12-35years who had no comorbidity. The subjects were enrolled from the Neurology out-patient department of Civil Hospital, Karachi. Healthy individuals matched for age, gender and body mass index were also enrolled as controls. Blood samples were collected between 8am and 10am for the evaluation of serum leptin levels using enzyme-linked immunosorbent assay. Data was analysed using SPSS 26. RESULTS: Of the 100 subjects, 64(64%) were males and 36(36%) were females. There were 50(50%) cases with mean age 19.34±5.65 years, and 50(50%) controls with mean age 19.76±5.67 (p=0.614). 100% of 50 individuals in control group did not have any family history of epilepsy,while 18% of cases (9) reported a positive family history. Mean leptin level in cases was 31.03±19.37 compared to 5.25±4.03 in the controls (p<0.05). CONCLUSIONS: Serum leptin levels were elevated in newly-diagnosed epileptic patients compared to healthy controls.

Correction: Evolution and consequences of individual responses during the COVID-19 outbreak.

[This corrects the article DOI: 10.1371/journal.pone.0273964.].

Iron/vanadium co-doped tungsten oxide nanostructures anchored on graphitic carbon nitride sheets (FeV-WO3@g-C3N4) as a cost-effective novel electrode material for advanced supercapacitor applications.

In this work, we studied the effect of iron (Fe) and vanadium (V) co-doping (Fe/V), and graphitic carbon nitride (g-C3N4) on the performance of tungsten oxide (WO3) based electrodes for supercapacitor applications. The lone pair of electrons on nitrogen can improve the surface polarity of the g-C3N4 electrode material, which may results in multiple binding sites on the surface of electrode for interaction with electrolyte ions. As electrolyte ions interact with g-C3N4, they quickly become entangled with FeV-WO3 nanostructures, and the contact between the electrolyte and the working electrode is strengthened. Herein, FeV-WO3@g-C3N4 is fabricated by a wet chemical approach along with pure WO3 and FeV-WO3. All of the prepared samples i.e., WO3, FeV-WO3, and FeV-WO3@g-C3N4 were characterized by XRD, FTIR, EDS, FESEM, XPS, Raman, and BET techniques. Electrochemical performance is evaluated by cyclic voltammetry (CV), galvanic charge/discharge (GCD), and electrochemical impedance spectroscopy (EIS). It is concluded from electrochemical studies that FeV-WO3@g-C3N4 exhibits the highest electrochemical performance with specific capacitance of 1033.68 F g-1 at scan rate 5 mV s-1 in the potential window range from -0.8 to 0.25 V, that is greater than that for WO3 (422.76 F g-1) and FeV-WO3 (669.76 F g-1). FeV-WO3@g-C3N4 has the highest discharge time (867 s) that shows it has greater storage capacity, and its coulombic efficiency is 96.7%, which is greater than that for WO3 (80.1%) and FeV-WO3 (92.1%), respectively. Furthermore, excellent stability up to 2000 cycles is observed in FeV-WO3@g-C3N4. It is revealed from EIS measurements that equivalent series resistance and charge transfer values calculated for FeV-WO3@g-C3N4 are 1.82 Ω and 0.65 Ω, respectively.

Depressed HCN4 function in the type 2 diabetic sinoatrial node.

Diabetic patients often have impaired heart rate (HR) control. HR is regulated both intrinsically within the sinoatrial node (SAN) and via neuronal input. Previously, we found lower ex vivo HR in type 2 diabetic rat hearts, suggesting impaired HR generation within the SAN. The major driver of pacemaking within the SAN is the activity of hyperpolarisation-activated cyclic nucleotide-gated 4 (HCN(4)) channels. This study aimed to investigate whether the lower intrinsic HR in the type 2 diabetic heart is due to changes in HCN4 function, protein expression and/ or distribution. The intrinsic HR response to HCN4 blockade was determined in isolated Langendorff-perfused hearts of Zucker type 2 Diabetic Fatty (ZDF) rats (DM) and their non-diabetic ZDF littermates (nDM). HCN4 protein expression and membrane localisation were determined using western blot and immunofluorescence, respectively. We found that the intrinsic HR was lower in DM compared to nDM hearts. The change in intrinsic HR in response to HCN4 blockade with ivabradine was diminished in DM hearts, which normalised the intrinsic HR between the groups. HCN4 protein expression was decreased in the SAN of DM compared to nDM controls with no change in the fraction of HCN4 localised to the membrane of SAN cardiomyocytes. The lower intrinsic HR in DM is likely due to decreased HCN4 expression and depressed HCN4 function. Our study provides a novel understanding into the intrinsic mechanisms underlying altered HR control in type 2 diabetes.

Evolution and consequences of individual responses during the COVID-19 outbreak.

In a long-lasting major disease outbreak such as that of COVID-19, the challenge for public health authorities is to keep people motivated and keen on following safety guidelines. In this study, a compartmental model with a heterogeneous transmission rate (based on awareness) is utilized to hypothesize about the public adoption of preventive guidelines. Three subsequent outbreaks in South Korea, Pakistan, and Japan were analyzed as case studies. The transmission, behavior change, and behavioral change ease rates of the disease were measured in these countries. The parameters were estimated using the maximum likelihood method with an additional identifiability analysis performed to determine the uniqueness of the estimated parameters for quantitatively comparing them during the first three waves of COVID-19. The mathematical analysis and simulation results show that individual responses had a significant effect on the outbreak. Individuals declining to follow the public health guidelines in Korea and Japan between the second and third waves contributed to making the third peak the highest of the three peaks. In Pakistan, however, individual responses to following public health guidelines were maintained between the second and third waves, resulting in the third peak being lower than the first, rather than being associated with the highest transmission rate. Thus, maintaining a high level of awareness is critical for containing the spread. Improvised public health campaigns are recommended to sustain individual attention and maintain a high level of awareness.

Pharmacological Validation for the Folklore Use of Ipomoea nil against Asthma: In Vivo and In Vitro Evaluation.

Oxidative stress is the key factor that strengthens free radical generation which stimulates lung inflammation. The aim was to explore antioxidant, bronchodilatory along with anti-asthmatic potential of folkloric plants and the aqueous methanolic crude extract of Ipomoea nil (In.Cr) seeds which may demonstrate as more potent, economically affordable, having an improved antioxidant profile and providing evidence as exclusive therapeutic agents in respiratory pharmacology. In vitro antioxidant temperament was executed by DPPH, TFC, TPC and HPLC in addition to enzyme inhibition (cholinesterase) analysis; a bronchodilator assay on rabbit's trachea as well as in vivo OVA-induced allergic asthmatic activity was performed on mice. In vitro analysis of 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH) expressed as % inhibition 86.28 ± 0.25 with IC50 17.22 ± 0.56 mol/L, TPC 115.5 ± 1.02 mg GAE/g of dry sample, TFC 50.44 ± 1.06 mg QE/g dry weight of sample, inhibition in cholinesterase levels for acetyl and butyryl with IC50 (0.60 ± 0.67 and 1.5 ± 0.04 mol/L) in comparison with standard 0.06 ± 0.002 and 0.30 ± 0.003, respectively, while HPLC characterization of In.Cr confirmed the existence with identification as well as quantification of various polyphenolics and flavonoids i.e., gallic acid, vanillic acid, chlorogenic acid, quercetin, kaempferol and others. However, oral gavage of In.Cr at different doses in rabbits showed a better brochodilation profile as compared to carbachol and K+-induced bronchospasm. More significant (p < 0.01) reduction in OVA-induced allergic hyper-responses i.e., inflammatory cells grade, antibody IgE as well as altered IFN-α in airways were observed at three different doses of In.Cr. It can be concluded that sound mechanistic basis i.e., the existence of antioxidants: various phenolic and flavonoids, calcium antagonist(s) as well as enzymes' inhibition profile, validates folkloric consumptions of this traditionally used plant to treat ailments of respiration.

Microscopic, phytochemical and antibacterial studies of Zizyphus oxyphylla Edgew. leaves.

Microscopic, phytochemical and pharmacological profiles are required for correct identification of a plant material to ensure consistent efficacy and safety. But such data are not available for the leaf of an important medicinal plant, Zizyphus oxyphylla Edgew. (Family: Rhamnaceae). Therefore, the current study aimed to investigate leaves of the plant for microscopic, phytochemical and antibacterial studies. Powdered material was subjected to microscopy, proximate analyses and estimation of total primary metabolites. Then, different types of extracts prepared using various solvents in order of increasing polarity were screened for antibacterial activity against seven standard strains. The most active extract was hydrolyzed and aglycone enriched fraction was extracted and screened for antibacterial activity. The powder microscopy indicated the presence of vascular bundles filled with cuboidal calcium oxalate crystals, anisocytic stomata and xylary vessels with reticulate and scalariform thickenings. Proximate features and primary metabolites provided characteristic identifying patterns. The most active extract (methanol) upon acidic hydrolysis exhibited higher activity against B. bronchiseptica (26.01±0.01 mm), S. aureus (26.00±0.00 mm), P. aeruginosa (24.03±0.02 mm) and M. luteus (24.02± 0.00 mm). The results of the current study provide identifying microscopic and phytochemical profiles that may be useful for correct identification of leaves of the plant. Aglycone enriched extract is having remarkable antibacterial activity hence may be used for activity-guided isolation.

Revamping of Chronic Respiratory Diseases in Low- and Middle-Income Countries.

Low- and middle-income countries (LMICs) endure an asymmetrically high burden of worldwide disease and death caused by chronic respiratory diseases (CRDs), i.e., asthma, emphysema, bronchiectasis, and post-tuberculosis lung disease (PTLD). CRDs are firmly related with indigence, infectious diseases, and other non-communicable diseases (NCDs) and add to complex multi-disease with great impact on the lives and livelihood of those affected. The pertinence of CRDs to health and demographic wellbeing is relied upon to increment in the long time ahead, as expectations of life rise and the contending dangers of right on time youth mortality and irresistible infections level. The WHO has distinguished the counteraction and control of NCDs as an earnest improvement issue and crucial for the sustainable development goals (SDSs) by 2030. In this review, we center on CRDs in LMICs. We examine the early life roots of CRDs, challenges in their avoidance, identification and administration in LMICs, and the pathways to resolve for accomplish valid widespread wellbeing inclusion.

Morphological, microscopic, and physicochemical studies of Diospyros montana.

Adulteration is the root cause of producing not only a chemically and pharmacologically inferior but also in some instances hazardous or poisonous drug. Despite availability of several techniques, microscopy and physicochemical analyses are the most practical approaches for crude drug authentication. Hence, the present study aimed to evaluate morphological, microscopic, and physicochemical properties of root, bark, leaf, and fruit of Diospyros montana Morphological properties were determined by sensory organs, whereas microscopic features of cross-sections and powders were determined by light and scanning electron microscopy. The proximate and fluorescence analyses were performed using the standard guidelines. The physical examination of fresh, shade-dried, and powdered material showed no significant change in color. The identifying cellular structures included cuboidal cork, pitted tracheids, scalariform, reticulate and spiral xylary vessels, and rosettes, raphide, and cuboidal calcium oxalate crystals. The stomatal number, stomatal index, vein-islet and vein-termination number, and palisade ratio in the leaf were found to be 293.91 ± 32.68 mm-2 , 64.18 ± 3.42%, 22.00 ± 3.81 mm-2 and 38.40 ± 5.81 mm-2 , and 3.85 ± 0.60, respectively. Total ash, acid insoluble ash, water soluble ash and sulfated ash of leaf (9.00 ± 0.50%, 1.67 ± 0.23%, 2.00 ± 0.22% and 14.50 ± 0.99%, respectively), foaming index of bark and root (111.11 ± 2.11), and swelling index of fruit (19.00 ± 3.45) were higher than the other parts. The powder of different parts showed characteristic colors in the daylight and UV light upon treatment with various regents. The plant was found to be rich in saponins, fibers, and flavonoids. The results of the present study may serve as identifiers of different parts of Diospyros montana.

Linking traditional anti-ulcer use of rhizomes of Bergenia ciliata (Haw.) to its anti-Helicobacter pylori constituents.

The rhizomes of Bergenia ciliata (B. ciliata, Family: Saxifragaceae) are widely used for treating gastric ulcers in folk medicine in Asia. It was hypothesized that anti-ulcer activity of B. ciliata is due to its anti-Helicobacter pylori (H. pylori) activity. The anti-H. pylori activity was investigated on six clinical bacterial isolates using agar well-diffusion and broth micro-dilution methods. The anti-H. pylori activity of amoxicillin (standard) was the highest (Zone of inhibition; ZI = 25 mm, minimum inhibitory concentration; MIC=0.125 µg/µL) whereas among all the extracts of the rhizomes, methanol extract showed the highest activity (ZI = 16 mm, MIC = 12.50 µg/µL). Bioassay guided isolation of methanol extract using chromatographic and crystallization techniques isolated bergenin (ZI = 21mm, MIC = 0.391µg/µL) as constituent responsible for anti-H. pylori activity. The present study describes for the first time anti-H. pylori activity and possible mechanism of anti-ulcer properties of rhizomes of B. ciliata.

Chemical and pharmacological comparison of modern and traditional dosage forms of Joshanda.

Recently, a traditional remedy (Joshanda) has been replaced largely by modern ready-to-use dosage forms, which have not been compared to the original remedy. Therefore, the present study aimed to compare a number of modern dosage forms with traditional remedy. Seven brands, 3 batches each, were compared with a Lab-made formulation with reference to analytical (proximate analyses, spectroscopic and chromatographic metabolomes) and pharmacological profiles (anti-inflammatory and antibacterial activities). Chemical and pharmacological differences were found between Lab-made Joshanda and modern dosage forms. Such variations were also found within the brands and batches of modern formulations (p < 0.05). The Lab-made Joshanda showed significantly higher pharmacological activities as compared to modern brands (p ). The results of the present study indicate that modern dosage forms are unstandardised and less effective than the traditional remedy. Characteristic profiles obtained from Lab-made Joshanda may be used as reference to produce comparable dosage forms.

Impact of Particle-Size Reduction on the Solubility and Antidiabetic Activity of Extracts of Leaves of Vinca rosea.

OBJECTIVES: The present study aimed to enhance the aqueous solubility of methanol extract of leaves of Vinca rosea (family: Apocynaceae) by particle-size reduction using milling and to evaluate its antidiabetic activity. MATERIALS AND METHODS: The methanol extract (ME) was micronized using a vibratory ball mill, operated at a vibratory speed of 15 Hz for 60 min at room temperature, and the resulting extract micronized ME (MME) was investigated to determine particle size, solubility, UV/visible profile, and in vitro antidiabetic activity. RESULTS: The average particle size of MME was 0.753±0.227 µm, which was less than half of that of the ME (2.007±0.965 µm). The solubility of MME was greater than that of the ME. MME exhibited 65.63%, 18.0%, and 96.87% higher antidiabetic activity in the glucose uptake by the yeast cells method, hemoglobin glycosylation assay, and the alpha amylase inhibition assay, respectively (p<0.05). CONCLUSION: The results of the present study indicate that micronization effectively enhanced the aqueous solubility and antidiabetic activity of methanol extract of leaves of Vinca rosea.

Development and validation of a stability-indicating RP-HPLC-FLD method for determination of 5-[(4-chlorophenoxy) methyl]-1, 3, 4-oxadiazole-2-thiol; A novel drug candidate.

The present study describes the development and validation of a simple high performance liquid chromatographic method for the determination of a novel drug candidate, 5-[(4-chlorophenoxy) methyl]-1, 3, 4-oxadiazole-2-thiol. The stability-indicating capacity of the method was evaluated by subjecting the compound's solution to hydrolytic, oxidative, photolytic, transition metal- and thermal- stress. The chromatographic separation was achieved over a C18 column (Promosil, 5 µm, 4.60 × 250 mm), maintained at 25°C, using an isocratic mobile phase comprising a mixture of acetonitrile and acidified water of pH 2.67 (1:1, v/v), at a flow rate of 1.00 mL/min and detection using a fluorescent light detector (excitation at 250 nm and emission at 410 nm). The Beer's law was followed over the concentration range 2.50-50.00 µg/ml. The recovery (98.56-100.19%, SD <5%), intraday accuracy and precision (97.31-100.81%, RSD <5%), inter-day accuracy and precision (97.50-100.75%, RSD <5%) and intermediate accuracy and precision (98.10-99.91%, RSD <5%) indicated that the method was reliable, repeatable, reproducible and rugged. The resolution and selectivity factors of the compound's peak from the nearest resolving peak, particularly in case of dry heat and copper metal stress, were found to be greater than 2 and 1, respectively, which indicated specificity and selectivity. The compound was extensively decomposed in alkaline-hydrolytic, oxidative, metal- and dry heat- stress. However, the compound in acidic and neutral conditions was resistant to photolysis. The results of the present study indicate that the developed method is specific, selective, sensitive and suitable, hence, may be used for quality control, stability testing and preformulation studies.

Decursin and decursinol angelate: molecular mechanism and therapeutic potential in inflammatory diseases.

Epidemiological studies have shown that inflammation plays a critical role in the development and progression of various chronic diseases, including cancers, neurological diseases, hepatic fibrosis, diabetic retinopathy, and vascular diseases. Decursin and decursinol angelate (DA) are pyranocoumarin compounds obtained from the roots of Angelica gigas. Several studies have described the anti-inflammatory effects of decursin and DA. Decursin and DA have shown potential anti-inflammatory activity by modulating growth factors such as vascular endothelial growth factor, transcription factors such as signal transducer and activator of transcription 3 and nuclear factor kappa-light-chain-enhancer of activated B cells, cellular enzymes including matrix metalloproteinases cyclooxygenase, and protein kinases such as extracellular receptor kinase, phosphatidylinositol-3-kinase, and protein kinase C. These compounds have the ability to induce apoptosis by activating pro-apoptotic proteins and the caspase cascade, and reduced the expression of anti-apoptotic proteins such as B-cell lymphoma 2 and B-cell lymphoma-extra-large. Interaction with multiple molecular targets and cytotoxic effects, these two compounds are favorable candidates for treating various chronic inflammatory diseases such as cancers (prostate, breast, leukemia, cervical, and myeloma), rheumatoid arthritis, diabetic retinopathy, hepatic fibrosis, osteoclastogenesis, allergy, and Alzheimer's disease. We have summarized the preliminary studies regarding the biological effects of decursin and DA. In this review, we will also highlight the functions of coumarin compounds that can be translated to a clinical practice for the treatment and prevention of various inflammatory ailments.

Ca(2+)-Clock-Dependent Pacemaking in the Sinus Node Is Impaired in Mice with a Cardiac Specific Reduction in SERCA2 Abundance.

BACKGROUND: The sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2) pump is an important component of the Ca(2+)-clock pacemaker mechanism that provides robustness and flexibility to sinus node pacemaking. We have developed transgenic mice with reduced cardiac SERCA2 abundance (Serca2 KO) as a model for investigating SERCA2's role in sinus node pacemaking. METHODS AND RESULTS: In Serca2 KO mice, ventricular SERCA2a protein content measured by Western blotting was 75% (P < 0.05) lower than that in control mice (Serca2 FF) tissue. Immunofluorescent labeling of SERCA2a in ventricular, atrial, sinus node periphery and center tissue sections revealed 46, 45, 55, and 34% (all P < 0.05 vs. Serca2 FF) lower labeling, respectively and a mosaic pattern of expression. With telemetric ECG surveillance, we observed no difference in basal heart rate, but the PR-interval was prolonged in Serca2 KO mice: 49 ± 1 vs. 40 ± 1 ms (P < 0.001) in Serca2 FF. During exercise, heart rate in Serca2 KO mice was elevated to 667 ± 22 bpm, considerably less than 780 ± 17 bpm (P < 0.01) in Serca2 FF. In isolated sinus node preparations, 2 mM Cs(+) caused bradycardia that was equally pronounced in Serca2 KO and Serca2 FF (32 ± 4% vs. 29 ± 5%), indicating no change in the pacemaker current, I f. Disabling the Ca(2+)-clock with 2 µM ryanodine induced bradycardia that was less pronounced in Serca2 KO preparations (9 ± 1% vs. 20 ± 3% in Serca2 FF; P < 0.05), suggesting a disrupted Ca(2+)-clock. Mathematical modeling was used to dissect the effects of membrane- and Ca(2+)-clock components on Serca2 KO mouse heart rate and sinus node action potential. Computer modeling predicted a slowing of heart rate with SERCA2 downregulation and the heart rate slowing was pronounced at >70% reduction in SERCA2 activity. CONCLUSIONS: Serca2 KO mice show a disrupted Ca(2+)-clock-dependent pacemaker mechanism contributing to impaired sinus node and atrioventricular node function.

Plant secretomics: identification, isolation, and biological significance under environmental stress.

Plant secretomes are the proteins secreted by the plant cells and are involved in the maintenance of cell wall structure, relationship between host and pathogen, communication between different cells in the plant, etc. Amalgamation of methodologies like bioinformatics, biochemical, and proteomics are used to separate, classify, and outline secretomes by means of harmonizing in planta systems and in vitro suspension cultured cell system (SSCs). We summed up and explained the meaning of secretome, methods used for the identification and isolation of secreted proteins from extracellular space and methods for the assessment of purity of secretome proteins in this review. Two D PAGE method and HPLC based methods for the analysis together with different bioinformatics tools used for the prediction of secretome proteins are also discussed. Biological significance of secretome proteins under different environmental stresses, i.e., salt stress, drought stress, oxidative stress, etc., defense responses and plant interactions with environment are also explained in detail.