Thiourea derivatives inhibit key diabetes-associated enzymes and advanced glycation end-product formation as a treatment for diabetes mellitus.

This study was designed to screen novel thiourea derivatives against different enzymes, such as α-amylase, α-glucosidase, protein tyrosine phosphatase 1 B, and advanced glycated end product (AGEs). A cytotoxicity analysis was performed using rat L6 myotubes and molecular docking analysis was performed to map the binding interactions between the active compounds and α-amylase and α-glucosidase. The data revealed the potency of five compounds, including E (1-(2,4-difluorophenyl)-3-(3,4-dimethyl phenyl) thiourea), AG (1-(2-methoxy-5-(trifluoromethyl) phenyl)-3-(3-methoxy phenyl) thiourea), AF (1-(2,4-dichlorophenyl)-3-(4-ethylphenyl) thiourea), AD (1-(2,4-dichlorophenyl)-3-(4-ethylphenyl) thiourea), and AH (1-(2,4-difluorophenyl)-3-(2-iodophenyl) thiourea), showed activity against α-amylase. The corresponding percentage inhibitions were found to be 85 ± 1.9, 82 ± 0.7, 75 ± 1.2, 72 ± 0.4, and 65 ± 1.1%, respectively. These compounds were then screened using in vitro assays. Among them, AH showed the highest activity against α-glucosidase, AGEs, and PTP1B, with percentage inhibitions of 86 ± 0.4% (IC50  = 47.9 µM), 85 ± 0.7% (IC50  = 49.51 µM), and 85 ± 0.5% (IC50  = 79.74 µM), respectively. Compound AH showed an increased glucose uptake at a concentration of 100 µM. Finally, an in vivo study was conducted using a streptozotocin-induced diabetic mouse model and PTP1B expression was assessed using real-time PCR. Additionally, we examined the hypoglycemic effect of compound AH in diabetic rats compared to the standard drug glibenclamide.

Electrolytes imbalance caused by amikacin in patients receiving multi drug resistance- tuberculosis treatment at Hazara region Kpk, Pakistan.

INTRODUCTION: Multi drug resistance-tuberculosis (MDR-TB) is considered amongst one of the fatal medical and public health issue. Greater frequency of adverse reactions has been observed using 2nd line antituberculosis drugs, which rises the rate of morbidity. Hypokalemia and hyponatremia are common in patients receiving MDR-TB treatment. MATERIALS AND METHODS: This study was aimed to assess the effect of amikacin as multidrug resistance tuberculosis treatment on serum sodium and potassium level. A total 179 subjects (82 males and 97 females) were screened within the period of one year for their serum electrolyte levels before and during the treatment. Their body weight and sociodemographic characteristics were also taken into considerations. Data were expressed in the form of mean ± standard error using One-way analysis of variance (ANOVA), followed by Dunnett's test. All the statistical analyses were performed using GraphPad Prism v. 6. RESULT: The results showed that amikacin caused a significant (p< 0.05) hypokalemia when used in combination with other drugs having no known effect on serum potassium level which proved to be the side effect of amikacin. The average serum potassium level in both intensive and continuation phase was significantly lower (p< 0.05) than the baseline (3.73 mmol/L), however, in the continuation phase the potassium level (3.58 mmol/L) was slightly higher than that of intensive phase (3.54 mmol/L). The average serum sodium level in both phases; intensive and continuation, remained in steady state (137.25 mmol/L and 137.87 mmol/L respectively) throughout the study period as compared to the base line (135.5 mmol/L). The amikacin drug lowered serum potassium level but had no effect on sodium level. CONCLUSIONS: It is concluded that the serum electrolytes should be monitored during the course of treatment and corrected accordingly.

Anti-Infective Potential Of A Medicinal Plant Pimpenella Stewartii Used Traditionally In Hazara.

BACKGROUND: Medicinal plants have been used since long as folk medicines and also in Ayuvedic and Unani systems of treatment. They have importance to save human and animal lives because of their utilization in the herbal medicines. In all over the word plants are in constant use for curing of various kinds of diseases. The herbal medicines are effective due to presence of bioactive compounds along with vitamins and minerals in them. Moreover, plants produce photo-chemicals mainly secondary metabolites which have significant pharmaceutical activities like antioxidant, antimicrobial, pesticidal and anti-tumour activity. They are very effective and have little or no side effects. METHODS: Five microbial strains including three fungal and two bacterial species were tested for the efficacy of plant materials by Agar diffusion method. The fungal species were Aspergilus flavus, Aspergilus niger and Alternaria alternata while bacterial species included Arvenia caratovora and Xanthomans spp. RESULTS: Results obtained for methanolic extracts of Pimpenella stewartii against Aspergilus flavus, Aspergilus niger and Alternaria alternata showed significant antifungal activity. The antibacterial screening of Methanolic extracts of Pimpenella stewartii were evaluated against Arvenia caratovora and Xanthomans spp. The best zone of inhibition was found against Xanthomans spp (97.33±2 mm) at 250 ppm concentration followed by Arvenia caratovora (49.7±14 mm) at same concentration.

EXPRESSION OF PEROXISOME-PROLIFERATOR ACTIVATED RECEPTORS-γ IN DIABETICS, OBESE AND NORMAL SUBJECTS.

BACKGROUND: Current researcn in type 2 diabetes mellitus focuses on the role of Peroxisome- Proliferator Activated Receptors (PPARs) in the pathogenesis of the Insulin Resistance Syndrome (IRS), which are pre-diabetic lesion and the hallmark of fully developed type 2 diabetes mellitus. This study aims at identifying the abnormal status of the PPAR-γ in adipose tissues of type 2 diabetes mellitus patients, when compared with matched normal controls. METHODS: This cross-sectional study was conducted in Ayub Medical College, Abbottabad, from 2012 to 2014. Sample included three equal groups of patients. Group-1 with diagnosed type 2 diabetes mellitus, aged 40-65 years, acting as the test group, Group-2 included non-diabetic obese, and Group-3 with normal subjects. Transcription Factor Assay for Peroxisome Proliferator Activated Receptor Gamma (gamma PPAR) was done on ELISA Technique from Nuclear Extract procured from Adipose Tissue of the subjects. RESULTS: Mean age of enrolled participants was 48.93 SD ± 6.52 years. Patients ranged between ages of 40 years to 67 years. The mean values of PPAR in normal, obese and diabetic group were 1.72 SD ± 0.28, 1.282 SE ± 0.18 and 1.283 SE ± 0.18 respectively. The difference in mean values of PPAR was significant p < 0.05: CONCLUSION: The levels of PPAR-γ in patients with type 2 Diabetes Mellitus and Obese cases are significantly lower than normal controls.

EFFECT OF HORMONAL CONTRACEPTIVES ON SERUM SEROTONIN IN FEMALES OF REPRODUCTIVE AGE GROUP.

BACKGROUND: Many types of hormonal contraceptives are in use nowadays for example oral pills, emergency contraceptive pills, vaginal rings, implantable rods and injectable contraceptives (combined and progestogens only). The purpose of this study was to determine and compare serum serotonin levels in married fertile females of reproductive age group using hormonal contraceptives with non-contraceptive users. METHODS: A total of 300 women were selected in the study. This cross sectional study included three groups; Group-1 (control), group-2 (combined oral contraceptive users) and group-3 (injectable contraceptive users). History and examination of subjects were recorded on pro forma. Levels of serum serotonin were measured using standard ELISA kits. Results were analysed by one way ANOVA and a p-value 0.05% was taken as significant, using SPSS 16.0. RESULTS: The mean age of the patients in group-1 was 30.4 ± 6.1 years, group-2 was 28.9 ± 4.9 and in group-3 was 2.5 ± 6.8 years. For subjects in group-1, group-2 and group 3 the mean ± SD concentration of serum serotonin was 160.68 ± 53.27 ng/dl, 227.3 ± 63.98 ng/dl and 118.19 ± 31.32 ng/dl. A significant (p = 0.00) difference was seen among three groups, i.e., group-1, group-2 and group-3. After applying Post HOC Tukey's HSD, there was statistically no significant difference between group-1 and group-2 (p = 0.956). Difference was seen between group-2 and group-3 (p = 0.00), it was also significant between group-3 and group-1 (p = 0.00). CONCLUSION: It was concluded that hormonal contraceptives affect the levels of serum serotonin.

Subclinical rickets.

OBJECTIVE: The study on sub clinical rickets is unique in the sense that it has not been preplanned conducted anywhere especially in Pakistan. The objective of present study was to explore the prevalence, gender and geographical distribution of sub clinical rickets and their related factors among school students. METHODS: Out of total participants, 189(90%) students were finally included in the study from rural, urban and suburban high schools of Hazara Division, KPK. The age of boys and girls students was 11 years to 16 years. Anthropometrics data along with daily intakes of meal and availability of sun shine was noted on record form. Sub clinical cases were diagnosed with abnormal biochemical findings without physical indications of rickets. RESULTS: Sub clinical rickets was found in 51(27%) students, out of which 15(8%) were boys and 36(19%) girls. Geographically, 26 cases of sub clinical rickets were from rural schools, 16 of urban and 09 found in suburban school. All sub clinical cases had serum level of sunshine vitamin D in between ≥18nmol/l to ≤39 nmol/l, but none of them had parathyroid level increased from upper normal range. Estimated quantities of vitamin D, calcium and phosphorus in daily intakes meal of boy's and girl's student were almost same and found less than recommended amount. CONCLUSION: Sub clinical rickets is camouflagic rickets among Hazarian school students, especially in girl gender. The major cause contributed to this problem is lack of synergistic effect of Sunshine Vitamin D.

Insulin deficiency among newly diagnosed type 2 diabetics.

BACKGROUND: Any patient above the age of 40 years, coming with the symptoms of diabetes is labelled as type 2 diabetic. If insulin levels are included in the protocol for initial investigations of diabetic patients, they can be differentiated as having insulin deficiency or insulin resistance. They can thus be treated accordingly. This study was conducted to see the prevalence of insulin resistance and insulin deficiency in newly diagnosed type 2 diabetics. METHODS: This study was conducted on 75 newly diagnosed diabetic subjects, and 75 control subjects for comparison. Fasting serum insulin was assayed by ELISA and HOMA-IR index was calculated. The diabetic subjects with fasting hyperglycaemia and serum insulin level below 20 microIU/ml and HOMA-IR index below 3.5 were grouped as insulin deficient (Group-A), and the diabetic subjects with fasting insulin level above 20 microIU/ml and HOMA-IR index above 3.5 were grouped as insulin resistant (Group-B). RESULTS: Twenty-eight percent subjects were found to have insulin level below 20 microIU/ml while 72% subjects had insulin resistance. When gender was taken into consideration, it was seen that 18.7% males had fasting insulin level of 6.98 +/- 0.737 microIU/ml and 9.3% females had fasting insulin level of 5.21 +/- 0.885 microIU/ml while 32% males and 40% females had insulin resistance. The mean age of male subjects with insulin resistance was significantly higher compared to the male subjects with insulin deficiency. Mean weight and body mass index of the male and female subjects having insulin resistance was significantly higher than their respective control groups and also higher than the subjects with insulin deficiency. Pearson coefficient of correlation was calculated for fasting serum insulin level with age and BMI. A significant positive correlation was observed between fasting serum insulin and age of females with insulin resistance. CONCLUSION: A considerable number of persons who develop diabetes after 40 years of age but are not insulin resistant. Twenty-eight percent subjects have relative insulin deficiency, and 72% subjects have insulin resistance.

Effect of body mass index on serum leptin levels.

BACKGROUND: Leptin is product of ob gene, an adipose tissue derived hormone that plays a key role in the regulation of body fat mass by regulating appetite and metabolism while balancing energy intake and energy expenditure. The objective of the study was to evaluate possible association between serum Leptin levels and Body Mass Index (BMI) of gender in adult age group. METHODS: Two-hundred-seventy subjects aged 20-50 years were randomly selected from general population of Abbottabad. After complete evaluation, demographic data was recorded and BMI calculated. The subjects were grouped on the basis on BMI. Non-fasting venous blood samples were drawn to measure serum Leptin and serum glucose levels. The data were analysed using SPSS-15. RESULTS: Serum Leptin levels and differences between genders were significant in all body mass indices. For normal BMI group the mean values for leptin were 2.6 +/- 1.5 etag/ml in men, and 17.3 +/- 10.2 etag/ml in women. For Group-2 mean leptin levels were 9.9 +/- 6.8 etag/ml in men, and 34.8 +/- 13.6 etag/ml in women. For Group-3 BMI comprising obese subjects mean values were 21.3 +/- 14.2 etag/ml in men, and 48.21 +/- 21.2 etag/ml in women (p<0.001). CONCLUSION: A progressive increase in serum leptin concentration was observed with an increase in BMI. Significant difference between leptin concentrations in either gender was found in normal, overweight and obese subjects.

Prevalence of Extended Spectrum ß-Lactamase and Antimicrobial Susceptibility Pattern of Clinical Isolates of Pseudomonas from Patients of Khyber Pakhtunkhwa, Pakistan.

Majority of gram negative pathogenic bacteria are responsible for extended spectrum ß-lactamases (ESBLs) production, which show resistance to some newer generation of antibiotics. The study was aimed at evaluating the prevalence of ESBL and antibiotic susceptibility pattern of Pseudomonas isolates collected during 2010 to 2014 from tertiary care hospitals of Peshawar, Pakistan. Out of 3450 samples, 334 Pseudomonas spp. isolates comprised of 232 indoor and 102 outdoor patients were obtained from different specimens and their susceptibility pattern was determined against 20 antibiotics. Antimicrobial susceptibility testing was carried out using the Kirby-Bauer agar diffusion method and ESBL production was detected by Synergy Disc Diffusion technique. The mean age group of the patients was 29.9 + 9.15 years. Meronem showed best activity (91.02%) from class carbapenem, ß-lactam and ß-lactamase inhibitors exhibited 69.16% activity, and doxycycline had a diminished activity (10.18%) to Pseudomonas spp. Outdoor isolates were more resistant than the indoor and during the course of the study the sensitivity rate of antibiotics was gradually reducing. ESBL production was observed in 44.32% while the remaining was non-ESBL. The moderate active antibiotics were amikacin (50.7%), SCF (51.4%), TZP (52.7%), and MXF (54.1%) among ESBL producing isolates. Lack of antibiotic policy, irrational uses (3GCs particularly), and the emergence of antibiotic resistant organisms in hospitals may be causes of high antibiotic resistance.

CCHF virus variants in Pakistan and Afghanistan: Emerging diversity and epidemiology.

BACKGROUND: Crimean Congo hemorrhagic fever (CCHF) has been reported from more than 30 countries in Africa, Asia, Eastern Europe and Middle East. The disease is considered endemic in Pakistan and neighboring countries like Iran and Afghanistan. OBJECTIVES: This study aimed to explore the genetic diversity of CCHF virus (CCHFV) detected in Pakistan and Afghanistan based on analysis of partial S-segment sequences. STUDY DESIGN: During 2011, one hundred samples satisfying the CCHF case definition were tested by (ELISA) and RT-PCR for detection of IgM antibodies and viral RNA, respectively. Phylogenetic analysis was carried out on partial S-segment nucleotide sequences using MEGA 5.0. RESULTS: Out of one hundred collected during 2011, 49 (49%) were positive for CCHF either by ELISA/RT-PCR or both. The mean age of the CCHFV positive cases was 30.32 years (range 18-56 years) and overall mortality rate was 20.4%. All CCHF virus isolates from this study clustered with strains previously reported from Pakistan, Iran and Afghanistan within the Asia-1 genogroup. Four distinct sub-clades were found circulating within Asia-1 genogroup. Six CCHFV strains found in Pakistan and Afghanistan grouped into a new sub-clade-D. CONCLUSIONS: Data from this study shows that endemic foci of CCHFV span the international border between Pakistan and Afghanistan with genetically diverse variants circulating in this region. Our findings emphasize to establish a laboratory based surveillance program and devise health policy measures to control CCHF infection especially in Baluchistan.

Subclinical nutritional rickets among adolescents in Kaghan Valley.

OBJECTIVE: To determine the occurrence of subclinical rickets and its causing factors among adolescent students of schools in Kaghan Valley, Pakistan. STUDY DESIGN: Observation cross-sectional study. PLACE AND DURATION OF STUDY: Department of Biochemistry and Health Sciences, Hazara University, Mansehra, Ayub Medical College and Teaching Hospital, Abbottabad, from March to April 2012. METHODOLOGY: Sixty seven students (34 boys and 33 girls) age between 11 - 16 years included in the study from different schools of Kaghan Valley, Pakistan. Characteristic, serum biochemical and nutritional status were measured for all the participants. On the basis of biochemical finding the boys and girls students were divided in to two groups, normal subjects and subclinical rickets (absent symptoms with altered biochemistry). RESULTS: Twenty six participants, 19 (73%) girls and 07 (27%) boys had biochemical abnormality but no clinical signs and symptoms of rickets. Low vitamin D and high alkaline phosphatase level were observed in 26 (100%), 21 (81%), low calcium in 17 (65%) and low phosphorus 7 (27%) subjects with subclinical rickets. None had high parathormone level above normal range. Nutritional intake of calcium, phosphorus and vitamin D was found less than the recommended daily intake in all the participants. CONCLUSION: Subclinical rickets is common problem among adolescent students especially in girls which is due to low nutritional intakes and avoidance of sunshine due to environmental and traditional impacts.