Prevalence and determinants of anemia among resident female university students from Southern Punjab, Pakistan.

Health and nutritional status of the women of reproductive age (WRA) is expected to be influenced with women literacy status alongside other sociodemographic indicators. However, data are scant to validate if literate young women have lower incidence of anemia prevalence. Nexus to the problem stated, a cross-sectional study was conducted on 1,541 female resident university students (FRUS) aged 17-35 years from a public sector university located in Southern Punjab, Pakistan. Hemoglobin (Hb) led screening for anemia was performed followed by nutritional assessment and structured questionnaire-based sociodemographic and dietary assessment. The data generated were analyzed using independent t-test, Chi-square, and response surface regression models. Response rate for the prevalence of anemia in FRUS was 38 percent with mean Hb levels 10.5 g/dL. With a significant effect (p = .001) of participants' weight on anemia prevalence, 22.45 percent of the sample population was recorded as underweight. Sociodemographic and dietary parameters analyzed suggested low daily food expenditure (x2 = 20.59; p = .000) and reduced intake of meat (x2 = 12.14; p = .01), beans & pulses (x2 = 18.56; p = .001) to significantly influence rate of anemia prevalence in FRUS. The study concludes high prevalence rate of anemia among FRUS to strongly relate with students' low monthly stipend, little daily food expenditure, and substandard dietary quality.

A study of association between presence or absence of GSTT1 and GSTM1 and/or single nucleotide polymorphism in FABP2 and GSTP1 with incidence of diabetes type 2: A case-control study.

OBJECTIVE: To assess the association of single nucleotide polymorphisms in fatty acid binding protein-2 (rs1799883) and glutathione S-transferase pi (rs1695) genes with presence/absence of glutathione S-transferase mu and glutathione S-transferase theta genes in type 2 diabetes. METHODS: The cross-sectional case-control study was conducted at Institute of Molecular Biology and Biotechnology during March till September 2019 and comprised type 2 diabetes patients and non-diabetic controls from two districts in southern Punjab. Polymerase chain reaction, polymerase chain reaction-restriction fragment length polymorphism and tetra-primer amplification refractory mutation system-polymerase chain reaction were applied to investigate glutathione S-transferase theta, mu and pi genes as well as fatty acid binding protein-2, as appropriate. The association of single nucleotide polymorphisms in all genes with the disease were studied either individually or in various combinations. Data was analysed using Minitab 18. RESULTS: Of the 448 subjects, 248(55.4%) were patients and 200(44.6%) were controls. Overall there were 213(47.5%) males and 235(52.5) were females, and 141(31.5%) were aged 30-46 years. The presence of rs1799883 in fatty acid binding protein-2 (p=0.03) and the absence of glutathione S-transferase mu gene (p<0.001) had significant association with type 2 diabetes, while the presence of glutathione S-transferase theta and rs1695 in glutathione S-transferase pi genes were not associated with the disease. Individuals with glutathione S-transferase mu gene null and Ileu/Ileu or Ileu/Val genotype of rs1695 in glutathione S-transferase pi gene have potential to develop type 2 diabetes in their lifetime (p<0.05). CONCLUSIONS: The presence of rs1799883 in fatty acid binding protein-2 and the absence of glutathione S-transferase mu gene were found to play significantly in the development of type 2 diabetes.

Exposure to variable doses of nickel oxide nanoparticles disturbs serum biochemical parameters and oxidative stress biomarkers from vital organs of albino mice in a sex-specific manner.

INTRODUCTION: This study was designed to report the biological effect of nickel oxide nanoparticles (NiO NPs) in albino mice. MATERIAL AND METHODS: Five weeks old albino mice of both sex were intraperitoneally injected either with 20 mg (low dose) or 50 mg/mL saline/kg body weight (high dose) of NiO NPs for 14 days. Saline-treated controls were maintained in parallel. Complete blood count, selected serum biochemical parameters and oxidative stress biomarkers from vital organs were determined in all subjects. RESULTS: Male mice treated with NiO NPS had increased blood urea nitrogen, elevated superoxide dismutase (SOD) in liver elevated MDA in liver, kidney and heart and reduced catalase activity in heart and kidney. Female mice treated with NiO NPs had significantly reduced serum albumin and total proteins, SOD in lungs and elevated MDA in liver. DISCUSSION: We are reporting that intraperitoneal injections of NiO NPs for 14 days drastically affect blood serum parameters and oxidative stress biomarkers from vital organs of albino mice. CONCLUSION: Toxic effects of NiO NPs were dose and sex dependent and they were more pronounced at higher dose and in male mice.

Single nucleotide polymorphism (rs7543472) in EPHA2 gene is associated with age-related cataract in subjects enrolled from Multan in southern Punjab: A case-control study.

OBJECTIVES: To examine whether Ephrin type A receptor 2 gene polymorphisms are associated with susceptibility to age-related cataract. METHODS: The case-control study was conducted from January to May 2014 in Multan, Pakistan, and comprised patients of age-related cataract enrolled from Nishtar Hospital, Multan, and age-matched healthy controls without any type of cataract from the local population. A questionnaire was used to gather clinical and epidemiological data. Deoxyribonucleic acid was extracted from blood samples, and analysis of rs11260867, rs3568293 and rs7543472 single nucleotide polymorphisms was performed by using tetra amplification-refractory mutation system polymerase chain reaction protocol. Data was analysed using SPSS 17. RESULTS: Of the 230 subjects, 129(%) were patients and 101(%) were controls. Among the three polymorphisms analysed, rs7543472 was associated with age-related cataract. Among the epidemiological and clinical factors, age, diabetes, blood pressure, smoking, radiation exposure, steroids usage and use of tranquilisers were associated with age-related cataract (p<0.05 each). CONCLUSIONS: Polymorphism rs7543472 was found to be associated with age-related cataract.

Association of single nucleotide polymorphism in CD28(C/T-I3 + 17) and CD40 (C/T-1) genes with the Graves' disease.

OBJECTIVE: To find out a correlation between the single nucleotide polymorphisms in cluster of differentiation 28 and cluster of differentiation 40 genes with Graves' disease, if any. METHODS: This case-control study was conducted at the Multan Institute of Nuclear Medicine and Radiotherapy, Multan, Pakistan, and comprised blood samples of Graves' disease patients and controls. Various risk factors were also correlated either with the genotype at each single-nucleotide polymorphism or with various combinations of genotypes studied during present investigation. RESULTS: Of the 160 samples, there were 80(50%) each from patients and controls. Risk factor analysis revealed that gender (p=0.008), marital status (p<0.001), education (p<0.001), smoking (p<0.001), tri-iodothyronine (P <0.001), thyroxin (p<0.001) and thyroid-stimulating hormone (p<0.000) levels in blood were associated with Graves' disease. CONCLUSIONS: Both single-nucleotide polymorphisms in both genes were not associated with Graves' disease, either individually or in any combined form.

Association of Single Nucleotide Polymorphisms in XRCC1 (194) and XPD (751) with Age-related cataract.

PURPOSE: Age-related cataract (ARC) is a multifactorial disease and different risk factors, including genetic and environmental, are responsible for the development of its various types. The aim of this study was to find out a correlation, if any, between ARC and the single nucleotide polymorphisms (SNPs) in DNA repair genes XRCC1 (X-ray repair cross-complementary-1) [Arg194Trp (rs1799782)] and XPD (xerodermapigmentosa complementation group D) [Lys751Gln (rs13,181]. METHOD: The genotype at these two SNPs was analyzed in 260 subjects (125 control and 135 patients) from Southern Punjab population (Pakistan) by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method. Genotype at both analyzed codons was correlated either individually or in various combinations with the studied epidemiological factors known to be associated with ARC. RESULTS: Our results indicated that both SNPs Arg194Trp in XRCC1 (P = 0.967) and Lys751Gln in XPD (P = 0.995) were not associated with ARC whether they were analyzed individually or in combined form (P > 0.05). Analysis of epidemiological factors revealed that age (P < 0.001), cast of subjects (P = 0.001), diabetes (P < 0.001), hypertension (P = 0.001), smoking habit (P = 0.01), drug abuse (P < 0.05), steroid use (P = 0.001) and body weight (P < 0.001) can influence the incidence of ARC in enrolled subjects. After applying Binary logistic regression it was found that the weight (P < 0.01), family history (P = 0.05), drug abuse (P = 0.05), smoking (P < 0.05) and steroid use (P < 0.05) has a significant association with the phenotype of the subjects. All epidemiological factors were also studied in association with various genotypic combinations of both SNPS, diabetes was the only factor that had a significant association (P < 0.001) association with ARC. Hypertension (P = 0.01), body weight (P < 0.05) and cast (P < 0.001) were found associated with ARC when epidemiological factors were individually correlated with ARC. Result of the two proportion test indicated that gender had no influence on the incidence of disease. CONCLUSION: It is concluded that studied SNPs in XRCC1 and XPD have no association with the incidence of age related cataract in the analyzed group of subjects.

Effect of Variable Doses of Zinc Oxide Nanoparticles on Male Albino Mice Behavior.

Zinc oxide nanoparticles (ZnO NPs) have diverse utility these days ranging from being part of nanosensors to be ingredient of cosmetics. Present study was designed to report the effect of variable doses of ZnO NPs on selected aspects of male albino mice behavior. Nano particles were synthesized by sol-gel auto-combustion method (Data not shown here). 10 week old male albino mice were divided into four experimental groups; group A, B and C were orally supplemented with 50 (low dose), 300 (medium dose) and 600 mg/ml solvent/kg body weight (high dose) of ZnO NPs for 4 days. Group D (control) orally received 0.2 M sodium phosphate buffer (solvent for ZnO NPs) for the same duration. A series of neurological tests (Rota rod, open field, novel object and light-dark box test) were conducted in all groups and performance was compared between ZnO NPs treated and control group. Muscular functioning during rota rod test was significantly improved in all ZnO NPs treated mice as compared to control group. While no significant differences in open field, novel object and light-dark box test performance were observed when data from studied parameters of specific ZnO NPs treatment were compared with the control group indicating that applied doses of ZnO NPs did not affect the exploratory, anxiolytic behavior and object recognition capability of adult male albino mice.

Effect of creatine monohydrate supplementation on learning, memory and neuromuscular coordination in female albino mice.

BACKGROUND: Research findings made over the last few years have highlighted the important role of creatine (Cr) in health and disease. However, limited information is available regarding the effect of Cr supplementation on cognation. Present study was designed to determine the effect of variable doses of Cr (1% and 3%) on selected parameters of female albino mice behaviour. METHODS: Following weaning, on 20th postnatal day, female albino mice were divided into three groups on the basis of dietary supplementation. Control group were was fed with normal rodent diet, whereas treated groups received diet supplemented with 1% and 3% Creatine monohydrate (Ssniff, Germany) for 10 weeks. Morris water maze (MWM), Rota rod and open field (OF) tests were carried out at the end of diet supplementation for neurofunctional assessment in all the groups. RESULTS: Data analysis showed that Cr supplementation did not affect the muscular activity and during rota rod test as well as locomotor and exploratory behaviour during OF test. Results of MWM probe trial indicated that mice supplemented with 3% Cr had significantly more entries in platform area than other two treatments (p=0.03) indicating improved spatial memory. Body weight remained unaffected (p>0.05) when compared between three experimental treatments. CONCLUSION: Female mice supplemented with 3% Cr showed improved spatial memory than mice fed on 1% Cr-supplemented diet and mice on normal rodent diet.

Serum levels of leptin, zinc and tryptophan in obese subjects with sleep deficits.

Obesity is an important risk factor for sleep disorders. This study aimed to evaluate the association of leptin, zinc and tryptophan (TRP) in obese subjects with sleep deficits [sleep apnea (SA), insomnia (IN)]. In this cross sectional case control, with the verbal and written consent 206, obese with sleep deficits and 30, non-obese/normal identified from various areas of Karachi, Pakistan. The socio-demographic data including; age, body mass index (BMI), education and residence, of participants was collected. After providing informed consent, fasting blood samples were taken and serum was collected. The serum concentration of leptin, zinc and TRP were analyzed by ELISA (Enzyme-linked immunosorbent assay), FAAS (Flame atomic absorption spectrophotometer) and HPLC (High performance liquid chromatography) respectively. A significant correlation was found between BMI (body mass index) and leptin, BMI and zinc, BMI and TRP. The correlation between leptin consecutively was significantly associated with zinc and TRP in obese patients. Sleep deficits elevated circulatory levels of leptin while lower zinc and TRP levels compared to levels seen in non-obese (Normal) subjects with no sleep deficits. Obese subjects exhibited significantly higher levels of leptin with sleep deficits compared with non-obese subjects with normal sleep pattern, while obese subjects with SA had significantly high levels of leptin than obese subjects with IN and IN+SA. Patients with sleep deficits had significantly lower levels of serum TRP and zinc than non-obese subjects with normal sleep pattern. Obese subjects with SA had significantly lower levels of zinc and elevated levels of TRP than obese subjects with IN. Obese patients with IN+SA had significantly lower levels of leptin and zinc than IN and SA , while TRP levels were significantly lower in subjects with IN than obese subjects with IN+SA and IN. These results suggest that elevated levels of leptin which are possibly by adiposity and lessened levels of zinc and TRP have a great impact on progression of obesity and their association can contribute to tempt sleep disorders.

Association of Polymorphisms (rs 1799782, rs25489 and rs25487) in XRCC1 and (rs 13181) XPD genes with Acute Coronary Artery Syndrome in Subjects from Multan, Pakistan.

Acute coronary artery syndrome (ACS) is the major cause of mortality in Pakistan with genetic and environmental influence on the incidence of the disease. This case-control study was designed to find out if a correlation is existing between ACS and single nucleotide polymorphisms (SNPs) in DNA repair genes XPD [at codon 751, rs 13181 (Lys to Gln)] and XRCC1 [at codon 399, rs25487 (Arg to Gln); 280, rs25489 (Arg to His) and 194, rs 1799782 (Arg to Trp)] either individually or in various combination with each other (haplotype analysis). The objective of this study was to find out the association of various studied risk factors and serum lipid profile of the subjects with the disease, if any. PCR-RFLP method was used to determine genotype at specific codon in 221 subjects (115 ACS patients and 106 healthy controls) from Southern Punjab population. Genotypic and allelic frequency distribution among the cases and controls revealed that all the studied SNPs were not individually associated with the ACS. Haplotype analysis revealed that subjects having wild type combination of all three XRCC1 SNPs had greater susceptibility to ACS than any other studied genotypic combinations. Analysis of risk factors revealed that hypertension (P<0.001), age (P=0.05), education (P<0.001), gender (P<0.001), family history (P=0.005), smoking habit (P=0.002) and diabetes (P<0.001) were significantly associated with the incidence of ACS. Serum lipid profile analysis indicated that cholesterol level was significantly higher (P=0.048) in patients (161.5mg/dL) than controls (142.1mg/dL) while triglyceride remained unaffected (P=0.87) when compared between the two treatments.

Gender-specific effects of CGP 55845, GABAB receptor antagonist, on neuromuscular coordination, learning and memory formation in albino mouse following neonatal hypoxia-ischemia insult.

GABAB receptor antagonists are experimentally proved as spatial memory enhancers in mouse models but their role has not been described following hypoxic-ischemic insult. 10-day-old albino mice were subjected to Murine model of hypoxia and ischemia. Following brain damage, mice were fed on normal rodent diet till they were 13 weeks old. At this time point, mice were divided into two groups. Group 1 received saline and group 2 received intraperitoneally CGP 55845 (1 mg/ml solvent/Kg body weight) for 12 days. Behavioural observations were made during rota rod, open field and Morris water maze test along with brain infarct measurement in both CGP 55845 treated and untreated groups. It was observed that application of GABAB receptor antagonist improved the over all motor function in male and female albino mice but effects were more pronounced in males. In open field, CGP 55845-treated female mice showed poor performance. CGP 55845 had no significant effect on learning and memory formation during Morris water maze test and also on brain infract size in both genders following hypoxia ischemia encephalopathy. Effects of CGP 55845 can be further explored in a dose and duration dependent manner to improve the learning and memory in albino mice following neonatal brain damage.

Effects of dietary creatine supplementation for 8 weeks on neuromuscular coordination and learning in male albino mouse following neonatal hypoxic ischemic insult.

Creatine monohydrate (Cr) is a dietary supplement known to improve cognitive functions and has positive therapeutic results under various clinical conditions. The aim of this study was to determine the effect of 2 % Cr supplementation on learning, memory formation, neuromuscular coordination, exploratory and locomotory in male albino mice following hypoxic ischemic insult. At postnatal day, 10 male albino mice pups were subjected to right common carotid artery ligation followed by 8 % hypoxia for 25 min. On postnatal day 20, male mice were separated from the litter and divided into two groups on the basis of special diet supplementation. One group was supplemented with 2 % Cr in diet while the other group was raised on ordinary rodent chow for 8 weeks. Behavioral observations were made during rota rod, open field and Morris water maze test for both treatments. It was observed that supplementation with 2 % Cr for 8 weeks following neonatal brain damage resulted in enhanced muscular strength, neuromuscular coordination and improved body weight. In Morris water maze test, it was observed that Cr supplementation significantly improved mean swimming speed and mice on 2 % Cr diet covered more distance but the spatial memory was not improved significantly following hypoxia ischemia encephalopathy (HIE). Open field parameters and percentage of infarct volume remained unaffected following Cr supplementation. We concluded that 2 % dietary Cr supplementation has a potential to improve the muscle strength and body weight in male albino mice following (HIE) and should be considered for the treatment of neurological ailments.

Effect of GABAB receptor antagonist (CGP35348) on learning and memory in albino mice.

The present study was designed to demonstrate the potential effect of CGP 35348 (GABAB receptor antagonist) on the learning, memory formation, and neuromuscular coordination in albino mouse. Mice were intrapertoneally injected with 1 mg CGP 35348/mL of distilled water/Kg body weight, while the control animals were injected with equal volume of saline solution. A battery of neurological tests was applied following the intrapertoneal injections. Results of rota rod indicated that CGP 35348 had no effect on neuromuscular coordination in both male (P=0.528) and female (P=0.125) albino mice. CGP 35348 treated females demonstrated poor exploratory behavior during open filed for several parameters (time mobile (P=0.04), time immobile (P=0.04), rotations (P=0.04), and anticlockwise rotations (P=0.038)). The results for Morris water maze (MWM) retention phase indicated that CGP 35348 treated male mice took shorter latency to reach the hidden platform (P=0.04) than control indicating improved memory. This observation was complemented by the swim strategies used by mice during training days in MWM as CGP 35348 treated males used more direct and focal approach to reach the platform as the training proceeded.

Effect of creatine monohydrate supplementation on various hematological and serum biochemical parameters of male albino mice following neonatal hypoxia-ischemia encephalopathy.

BACKGROUND: Present study was designed to report the effect of 2% creatine monohydrate supplementation for 8, 12 and 15 weeks on hematology and serum biochemical profile of male albino mouse following hypoxic ischemic insult on postnatal day 10. METHODS: 66 Blood samples (2% creatine monohydrate supplemented (N = 34) and unsupplemented (N = 32)) were analyzed for various hematological (blood glucose, packed cell volume, total WBC count, total RBC count) and serum biochemical parameters (cholesterol, AST, ALT, HDL, LDL, total protein, triglycerides). RESULTS: ALT had higher concentrations in mice feeding on normal diet for 8 (P > 0.01) and 12 weeks (P > 0.01) following asphyxia and in 12 weeks treatment without asphyxia (P = 0.006) when compared with the creatine supplemented mice. LDL (P = 0.011) and cholesterol (P > 0.01) had higher concentrations in mice on normal diet for 12 weeks following hypoxia ischemia. Cholesterol (P > 0.01) in 12 and glucose (P = 0.006) in 15 week treatment group had significantly lower concentrations in creatine supplemented male albino mice when compared with untreated group following hxpoic-ischemic insult. CONCLUSION: We concluded that creatine supplementation following hypoxic ischemic insult helps in maintain the normal blood chemistry.

Association of GSTTI, M1 and Polymorphism in GSTPI with Chronic Periodontal Disease in a Pakistani Population.

OBJECTIVE: Chronic periodontal disease (CP) is a multifactorial infectious and inflammatory disease that occurs due to the challenge between the immune response of the host and specific periodontal bacteria, and that can lead to tooth loss due to damage inflicted to the supporting tissue. The current study investigates the genotypes of the GSTM1 and GSTT1 genes, along with the allelic frequency of the single nucleotide polymorphism [SNP; rs1695] in the GSTP1 gene and correlates them individually or in various combinations with the incidence of CP. METHODS: A total of 203 clinically confirmed CP patients and 201 control subjects were enrolled from Multan and Dera Ghazi Khan Districts in Pakistan from April to July 2022. Multiplex Polymerase Chain Reaction (PCR) and tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) approaches were applied to determine the genotypes of the studied GSTs. The association of rs1695 in GSTP1 with CP was studied both individually and in various combinations with GSTM1 and T1. RESULTS: The absence of GSTM1, the presence of GSTT1 and the presence of the mutant allele (G) at rs1695 in GSTP1 were found to be significantly associated with CP. Patients aged between 10 and 30 years were more affected by CP. CONCLUSION: Our results indicate that the genotypes of the analyzed GSTs affect the levels of protection from oxidative stress and may therefore influence the disease progression in CP.

Machine learning methods for predicting major types of rheumatic heart diseases in children of Southern Punjab, Pakistan.

Objective: Rheumatic heart disease (RHD) is a major health problem in the world, particularly in developing countries. This study aimed to predict mitral regurgitation (MR) and mitral stenosis (MS) RHD among children with RHD. Methodology: Data was collected from the Pediatric Cardiology Department at Chaudhry Pervaiz Elahi Institute of Cardiology Multan, Pakistan from March to October 2019. A sample of 561 children aged 4-14 years, who were diagnosed with RHD of either MR or MS, were recruited from the hospital's outpatient department. The presence of multivariate outliers was detected, and different machine learning methods, including subset logistic regression, subset logistic regression after deletion, stepwise winsorized logistic regression, robust logistic regression, subset deep neural network, and random forest models were compared using the area under receiver operating characteristics (ROC) curve, sensitivity, and specificity. Parsimony was also considered in model selection. Results: Out of 561 patients in this study, 75.94% had RHD MR and 24.06% had RHD MS. The average age of study participants was 9.19 ± 2.45 years and of them 55.43% were male. Among the male participants, 58.6 and 45.2% had MR and MS, respectively; and among female participants, those were 70.4 and 29.6%, respectively. Subset logistic regression after deletion appeared as competitive with a discrimination power of 90.1% [95% CI 0.818-0.983]. The sensitivity and specificity of this model were 85.1 and 70.6%. Conclusion: The best predictive model was subset logistic regression after deletion. The predicted method will be used in the decision-making process, which helps early diagnosis of the disease and leads to prevention. The study findings provide the proper guideline for earlier diagnosis of the RHD MR and MS cases among children with RHD in Pakistan.

Establishing growth charts for proposed body shape and size index of the Pakistani population, using quantile regression approach.

BACKGROUND: Obesity leads to other fatal diseases like diabetes, cardiovascular diseases, depression, and some forms of cancer. Still, the well-known tool to measure obesity is the body mass index. But it usually failed in the measurement of adipose tissues. So, we present a novel anthropometric measure, called body shape and size index which is developed by the combination of major anthropometric determinants: body surface area, body mass index, weight, and height. METHODS: This study is based on cross-sectional data consisting of 7224 individuals that were taken from the city Multan, Punjab, Pakistan. All the individuals, both males, and females, of age 2 years and above were included in the study except the pregnant women. The variables included in this study are gender, area (urban and rural), age (years), weight (kg), and height (meters). Growth charts of quantile regression are used for the inferential analysis of data. Comparison of proposed body shape and size index at different obesity levels has also been made to access the relationship of proposed body shape and size index with obesity. RESULTS: The results show that the proposed body shape and size index has a great association with body surface area, body mass index, weight, height, and age. The proposed body shape and size index has a high negative association with body surface area, moderate negative association with body mass index and weight, and low negative association with height and age. According to growth charts of body shape and size index, after the age of 25 years, body shape and size index curves go upward while it smoothly goes downward at the age of 50 years but decreases in earlier ages. Body shape and size index showed a significant association with body shape and body size (body development) at the same time. CONCLUSION: Body shape and size index is found, generally linear with age, and increased with decreasing body mass index and body surface area. The proposed index has an indirect relationship with obesity. Body shape and size index with low values indicates a high risk of obesity. While, however, body shape and size index with high values indicates a low risk of obesity. Applications of the proposed body shape and size index are also presented in statistical modeling.

Cutaneous Leishmaniasis in the Metropolitan City of Multan, Pakistan, a Neglected Tropical Disease.

Cutaneous leishmaniasis is a vector-borne infection caused by flagellated parasitic protozoans in the genus Leishmania. Promastigotes are the infective stage, which are transmitted by the bite of female sand flies of the genera Phlebotomus or Lutzomyia. Three clinical forms of Leishmania infection are recognized in humans, and they are caused by different Leishmania species. They are cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (ML), and visceral leishmaniasis (VL). Up to 2 million cases of leishmaniasis are reported annually worldwide. In Pakistan, CL is widely distributed in endemic areas. In the current study, a survey was conducted to investigate the occurrence of CL in an endemic area of the metropolitan city of Multan, Pakistan. The material for diagnosis of CL was obtained from 461 patients from 239 clinical laboratories and health care centers in Multan. Of these patients, 42.9% (198 of 461) having positive findings of amastigotes in Giemsa stained tissue impression smears. The distribution of CL skin lesions was highly variable. Most patients (76.2%) had a single lesion. Most CL lesions were found on a single hand (23.7%). The prevalence of two CL lesions per patient was 21.7%, while three or more positive CL lesions were observed in 2.0% of patients. The 198 CL-positive patients ranged in age 10-40 yr old, 101 were males, and 97 were females. Logistic regression analysis of leishmaniasis versus age and gender showed that both the age and gender had a significant (P < 0.05) effect on the occurrence of infection. Outbreaks of CL were reported in 14 of the 68 Union Councils of Multan.

Creatine monohydrate supplementation for 10 weeks mediates neuroprotection and improves learning/memory following neonatal hypoxia ischemia encephalopathy in female albino mice.

Currently there are no uniform standard treatments for newborn suffering from cerebral hypoxia-ischemia (HI) and to find new and effective strategies for treating the HI injury remains a key direction for future research. Present study was designed to demonstrate that optimal dose (1 or 3%) of creatine monohydrate (Cr) for the treatment of neonatal HI in female albino mice. On postnatal day 10, animals were subjected to left carotid artery ligation followed by 8% hypoxia for 25 minutes. Following weaning on postnatal day 20, mice were divided into three treatments on the basis of diet supplementation (Normal rodent diet, 1% and 3% creatine supplemented diet) for 10 week. A battery of neurological tests (Rota rod, open field and Morris water maze) was used to demonstrate effect of Cr supplementation on neurofunction and infarct size following HI. Open field test results indicated that Cr supplementation had significantly improved locomotory and exploratory behavior in subjects. It was observed that Cr treated mice showed better neuromuscular coordination (rota rod) and improved spatial memory (Morris Water Maze test). A significant affect of creatine supplementation in reducing infarct size was also observed. Post hoc analysis of post hoc multiple comparisons revealed that mice supplemented with 3% Cr for 10 weeks performed better during Morris water maze test while 1% Cr supplementation improved the exploratory behavior and gain in body weight than control group indicating that Cr supplementation has the potential to improve the neurofunction following neonatal brain damage. This article is part of a Special Issue entitled SI: Brain and Memory.