In vitro comparative evaluation of Tamarix gallica extracts for antioxidant and antidiabetic activity.

Tamarix gallica known as Jhau is traditionally used as expectorant, liver tonic, laxative, astringent, and antidiarrheal. The current study was proposed to determine the in vitro antioxidant, antidiabetic properties of the methanolic, ethanolic, and aqueous extracts of Tamarix gallica arial part, subsequently the phytochemical evaluation. Hence, Tamarix gallica arial part extracts were extracted with methanol (MthTg), ethanol (EthTg), and distilled water (AqTg). Extracts phytochemical analysis were accomplished to identify the phenolic components (TPC and TFC). Extracts antioxidant property was evaluated by DPPH, FRAP, and ABTS assay. For antidiabetic property, α-amylase and α-glucosidase inhibitory activities were assessed. One Way ANOVA was applied statistically by means of software SPSS Statistics 23 and attained data were definite as mean ± standard deviation. Result revealed that EthTg have the maximum TPC and TFC levels than MthTg and AqTg. Antioxidant property in relations of DPPH (lowest IC50 = 1.309 ± 0.31), FRAP (323.51 ± 2.32), and ABTS (266.97 ± 25.14) assay was also highest in EthTg. EthTg was also exposed highest α-amylase and α-glucosidase inhibition activity with lower IC50 (1.116 ± 0.051; 0.402 ± 0.2, respectively). The extracts antioxidant and antidiabetic activities order was as EthTg > MthTg > AqTg. TFC and TPC also revealed directly proportional correlation with antioxidant, and antidiabetic properties of the Tamarix gallica arial part extracts. Results noticeably stated that the ethanolic extract of Tamarix gallica have the highest antioxidant and antidiabetic properties. Tamarix gallica has competency to reduce the oxidative stress and can be utilized in the management of diabetes.

The Hepatoprotective and Anti-Nephrotoxic Potential of Methanolic Extract of a Polyherbal Preparation in CCl4-Induced Liver Injury Model of Wistar Rats.

The liver and kidneys are the vital organs of the body and perform important life-sustaining functions in the body. Synthetic drugs used in the treatment of liver and kidney diseases are sometimes inadequate and can lead to serious side effects. Medicinal herbs and plants were used to combat diseases for a long time and combination therapy is preferred over single plant therapy. In the current study, the Asparagus racemosus, Mucuna pruriens, Anacyclus pyrethrum, and Tribulus terrestris polyherbal preparation (PHP) was selected to evaluate its hepatoprotective, antioxidant, and anti-nephrotoxic potential. The methanolic extract of PHP was prepared following standard protocols. Fifty-six albino rats were divided into 7 groups (n = 8). The negative control (NC) having the healthy rats and the remaining 6 groups were induced liver toxicity by intraperitoneally injecting 0.5 mL/kg of 50% CCl4 in olive oil. Group 2 was positive control and group 3 and 4 received silymarin standard drug at the dose of 100 and 200 mg/kg body weight. Groups 5, 6, and 7 (PHP-1, PHP-2, PHP-3) were the liver-damaged rats receiving the PHP at a dose of 50, 100, and 150 mg/kg body weight. Blood samples were collected at 21 of the trial, to evaluate oxidative stress, hepatoprotective and anti-nephrotoxic potential. Results of liver function tests revealed significant (P < .05) hepatoprotective activities of PHP after intoxication with CCl4 of albino rats as compared to standard groups. Moreover, results of renal functions also showed that PHP has a significant (P < .05) restoring the capacity of blood urea, creatinine, and uric acid in intoxicated rats as compared with the control group. The PHP also reduced the oxidative stress in the treatment groups by increasing the total antioxidant capacity and reducing the total oxidative status. It can be concluded that selected medicinal plants have a potential role in the management of liver and kidney disorders. So, by running the clinical trial on a large scale and by isolating the phytochemical constituents responsible for hepatoprotective and nephroprotective activities, locally prepared drugs could be developed to manage liver and renal disorders.

Anti-diabetic Potential of Indigenous Medicinal Plants of Cholistan Desert, Pakistan: A Review.

Cholistan Desert is a sandy desert located in southern Punjab, Pakistan. The area is rich in more than 64 medicinal plants among 138 plant species. It is noteworthy that this remote desert lacks modern health care facilities and its inhabitants are dependent on locally-available plant species for the treatment of acute and chronic illnesses. Medicinal plants, traditionally have been ideal sources of remedies for the management of many non-communicable diseases; most modern prescriptions drugs have their origins from plants. Diabetes is increasing at an alarming rate in the past few decades. Whereas medicinal plants are used globally, the specific properties of only a few have been identifies scientifically. Similarly, little scientific evidence exists that confirms the efficacy of the medicinal plants of this region for diabetes management. Ethnobotanical studies show that locally-available medicinal plants do have anti-diabetic potential. We reviewed the medicinal properties of 36 of these plants. Several ingredients derived from these plants have chemical constituents that demonstrate anti-diabetic activity, thereby validating their importance for the management of diabetes.

Epidemiologic profile of hard ticks and molecular characterization of Rhipicephalus microplus infesting cattle in central part of Khyber Pakhtunkhwa, Pakistan.

Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were examined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan.

Phytochemical Screening, Antioxidant, and Inhibition Activity of Picrorhiza kurroa Against α-Amylase and α-Glucosidase.

Picrorhiza kurroa (P.K) usually familiar as kutki is a well-known plant in the Ayurvedic system of medicine due to its reported activities including antidiabetic, antibacterial, antioxidant, antitumor, anti-inflammatory, and hepatoprotective. The current research was intended to evaluate the antioxidant, inhibition activity of the ethanolic, methanolic, and aqueous extracts of P.K roots against α-amylase and α-glucosidase in vitro, after the phytochemical analysis. For this purpose, P.K roots were extracted with ethanol (EthPk), methanol (MthPk), and distilled water (AqPk) and phytochemical study of the extracts were performed to recognize the total phenolic content (TPC) and total flavonoids content (TFC). Antioxidant capability of the extracts was assessed by FRAP, ABTS, and DPPH assay. α-amylase inhibitory and α-glucosidase inhibitory activities were also determined. Software SPSS-23 was used to statistically analyze with One Way ANOVA and results were stated as mean standard deviation. Result of the study showed that MthPk contained the maximum concentration of TPC and TFC than EthPk and AqEh. Antioxidants in terms of DPPH (lowest IC50 = .894 ± .57), FRAP (612.54 ± 11.73) and ABTS (406.42 ± 4.02) assay was also maximum in MthPk. MthPk was also showed maximum inhibition activity against α-amylase and α-glucosidase with lowest IC50 (.39 ± .41; .61 ± .24), respectively. The extracts α-amylase and α-glucosidase inhibitory activities order was as MthPk > EthPk> AqPk. Results clearly specified that the methanolic extract of Picrorhiza kurroa have the maximum antioxidant, α-amylase, and α-glucosidase inhibitory activities. A positive correlation of TPC, TFC with antioxidant, and α-amylase and α-glucosidase inhibition activities of the P.K roots were also shown. The plant has capability to diminish the oxidative stress and can be used to treat diabetes by inhibiting α-amylase and α-glucosidase actions.

Changes in the Prevalence of Natural Paramphistomum Cercariae Infection in Indoplanorbis and Lymnaea Intermediate Hosts Influenced by Meteorological Factors.

Paramphistomosis is a neglected ruminant parasitic disease caused by trematodes known as Paramphistomum, which has a diheteroxenic life cycle involving freshwater snail genera, i.e., Planorbis and Lymnaea as the intermediate host and mammals as the definitive host. Snail vector distribution, infection with Paramphistomum spp. cercariae, preferred habitat, and their relationship with certain meteorological factors were not investigated in the province Khyber Pakhtunkhwa of Pakistan. Therefore, this study is designed to evaluate the effects of meteorological factors on the occurrence and severity of Paramphistomum spp. cercariae in Indoplanorbis and Lymnaea intermediate snail hosts. For this purpose, a cross-sectional survey was conducted from October 2018 to September 2019, and snails were collected and then identified using snail shell morphology; their infection with Paramphistomum spp. cercariae was determined through microscopy; and descriptive statistics were used to estimate the prevalence of infection and evaluate their occurrence relationship with a certain meteorological factors including temperature, humidity, rainfall, and pan evaporation in different districts of the above-mentioned province of Pakistan, i.e., adjacent areas of Bara and Kabul rivers in district Nowshehra, Kabul River (Sardaryab) of district Charsadda, Kalpani River of district Mardan, and Indus River (Hund) of district Swabi. A total of 2,706 Indoplanorbis (1539) and Lymnaea (1167) snails were collected, in which overall 10.30% shed Paramphistomum spp. cercariae. The highest infection rate was found in the river of district Swabi (13.20%), while the lowest in adjacent rivers of district Nowshehra (8.19%). Meteorological factors play an essential role in the causation of Paramphistomum spp. infection, parasitic reproduction, vector growth, and survival. Due to these factors, high significant prevalence was found in the summer season (11.83), followed by autumn (11.25), which might be due to optimum temperature, relative humidity, and rainfall (p < 0.05). It was concluded from the study that meteorological factors contribute to the prevalence of Paramphistomum species in the Indoplanorbis and Lymnaea, which act as vectors for the disease paramphistomosis, which may lead to the increased intensity of infection outbreaks of the parasite population in humans and domestic animals.

Genetic Diversity of Polymorphic Marker Merozoite Surface Protein 1 (Msp-1) and 2 (Msp-2) Genes of Plasmodium falciparum Isolates From Malaria Endemic Region of Pakistan.

Background: Understanding the genetic diversity of Plasmodium species through polymorphic studies can assist in designing more effective control strategies of malaria like new drug formulation and development of a vaccine. Pakistan is moderate endemic for Plasmodium falciparum, but little is known about the genetic diversity of this parasite. This study aimed to investigate the molecular diversity of P. falciparum based on msp-1 and msp-2 genes in the malaria-endemic regions of Khyber Pakhtunkhwa, Pakistan. Methods: A total of 199/723 blood samples, tested positive by microscopy for falciparum malaria, were collected from four districts (Dera Ismail Khan, Karak, Mardan, and Peshawar) of Khyber Pakhtunkhwa. Nested PCR amplification technique was employed to target block 2 of msp-1 and the central domain of msp-2 genes, including their respective allelic families K1, MAD20, RO33, FC27, and 3D7/IC, and to detect the extent of genetic diversity of P. falciparum clinical isolates. Results: Among the 199 microscopy-positive P. falciparum samples, a total of 192 were confirmed using PCR. Ninety-seven amplicons were observed for msp-1 and 95 for msp-2. A total of 33 genotypes, 17 for msp-1 (eight K1, six MAD20, and three RO33) and 16 for msp-2 (nine FC27 and seven 3D7/IC), were identified. The specific allelic frequency of the K1 family was higher (44.3%) than that of MAD20 (33.0%) and RO33 (23.0%) for msp-1, while the FC27 allelic family was dominant (60.0%) compared with 3D7/IC (40.0%) for msp-2. No polyclonal infection was observed in msp-1 and msp-2. The expected heterozygosity was 0.98 and 0.97 for msp-1 and msp-2, respectively. Conclusion: It was concluded that the P. falciparum populations are highly polymorphic, and diverse allelic variants of msp-1 and msp-2 are present in Khyber Pakhtunkhwa, Pakistan.

Epidemiology and molecular characterization of Theileria annulata in cattle from central Khyber Pakhtunkhwa, Pakistan.

Theileria annulata is a tick-borne hemoprotozoan parasite responsible for tropical theileriosis in the bovine population, which causes substantial economic losses to the livestock sector. The present study has investigated, characterized, and shaped epidemiologic and phylogenetic profiles of T. annulata infection in the cattle population of central Khyber Pakhtunkhwa, Pakistan. A total of 600 blood samples were collected from cattle. Microscopy and PCR (18S rRNA taxonomic marker) assays were performed to detect T. annulata infection in cattle from the study area. The overall relative prevalence rates of T. annulata in the examined cattle population were 12.8% (microscopy) and 23.7% (PCR). District-wise analysis (microscopy/PCR) showed that cattle from district Mardan were found more infected (16.0%/28.0%), as compared to cattle from district Charsadda (13.5%/25.5%) and district Peshawar (9.0%/17.5%). Based on host demographic and ecological parameters analysis, theileriosis was found to be higher in young, female, crossbred, freely grazing, tick-infested, and irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, tick infestation, acaricides use, and feeding method were significant risk factors (P<0.05) whereas multivariate analysis indicated that host age, gender, tick infestation, acaricidal application, and feeding method were potential risk factors (P<0.05) for tropical theileriosis in the cattle population. Phylogenetic and sequence analysis showed that T. annulata 18S rRNA isolates shared homology and phylogeny with other isolates from Asia and Europe. This study has addressed the epidemiology and phylogeny of T. annulata circulating in bovid in the study area where gaps were still present. These findings will serve as a baseline and will facilitate future large-scale epidemiological investigations on tropical theileriosis in the cattle population at a national level.

A cross-sectional survey of hard ticks and molecular characterization of Rhipicephalus microplus parasitizing domestic animals of Khyber Pakhtunkhwa, Pakistan.

BACKGROUND: In tropical and subtropical countries, tick infestation causes major public health problems and considerable financial losses to the livestock industry. This study was aimed to assess the species composition of richness and analyze the phylogeny of Rhipicephalus microplus in the District Bannu of Khyber Pakhtunkhwa, Pakistan. METHODS: Collected ticks were identified morphologically and DNA extracted from R. microplus was amplified and subjected to sequencing. RESULTS: A total of 3,600 animals were examined among them 1,494 animals were found to be infested with ticks, including 669 cows, 476 buffaloes, 163 goats, and 186 sheep (p = 0.001). Tick infestation was significantly high (43.58%) in animals of age group (<1 year) (p-value = 0.027). Female animals were more (44.05%) infested with ticks than males (34.43%) (p = 0.001). The intensity of infestation was significantly higher in summer (77.49%) (p = 0.001). A total of 5,557 ticks were collected comprising three genera and six species. R. microplus was predominantly prevalent (n = 1,474; 26.52%), followed by Rhipicephalus annulatus (n = 1,215; 21.86%), Hyalomma anatolicum (n = 1,139; 20.49%), Hyalomma marginatum (n = 1,086; 19.54%), and Rhipicephalus turanicus (n = 761; 13.69%), while the least common was Haemaphysalis aciculifer (n = 80; 1.43%) (p = 0.001). Morphologically identified R. microplus species were also analyzed genetically by using two genetic markers 16S ribosomal RNA (16S rRNA) and internal transcribed spacer 2 (ITS2) genes. The phylogenetic study revealed that R. microplus is genetically diversified and clustered in clade B with R. microplus species from China, India, and Pakistan. CONCLUSION: Ticks infestation was significantly correlated with various factors including age, sex, season, and animal type. R. microplus genetically resembled species reported from India and China. However, major knowledge gaps concerning various species of ticks exist and many areas are still unexplored in Pakistan. Therefore, it is necessary to explore the epidemiological and molecular aspects of various tick species in other regions of southern Khyber Pakhtunkhwa.

Assessment of ameliorative effect of Trigonella foenum-graecum against CuO-NPs induced toxicity in Oreochromis mossambicus.

The current study assessed the ameliorative effect of Trigonella foenum graceum extract against copper oxide nanoparticles (CuO-NPs) induced toxicity in Oreochromis mossambicus. For this purpose 100 healthy fish weighing 20±2.34g were randomly divided into five different groups in duplicates and designated as control (C) no treatment, positive control (G*) treated with 0.12mg/L of CuO-NPs, experimental co-treated groups G1, G2 and G3 were treated with Trigonella foenum-graecum extract @ 18, 26 and 52mg/L along with 0.12 mg/L of CuO-NPs, respectively. In this study significant (P<0.05) changes were observed in the antioxidant activity of enzymes and histological alterations in the liver and intestine of fish in G*, G1 and G2 groups while a good ameliorative response of Trigonella foenum-graecum was observed in G3. Dose dependent alterations in glutathione, lipid peroxides, catalase, and malondialdehyde as well as histological architecture of liver and intestine were observed in treated groups, where more alterations were observed in positive control and low dose treated groups of Trigonella foenum-graecum. Moreover, more ameliorative effect was observed in high dose of Trigonella foenum-graecum treated group (G3). This study is novel as no previous data is available on the amelioration of Trigonella foenum-graecum extract against CuO-NPs induced toxicity in fish.

An Assessment of the Molecular Diversity of Ticks and Tick-Borne Microorganisms of Small Ruminants in Pakistan.

This study investigated ticks and tick-borne microorganisms of small ruminants from five districts of the Federally Administered Tribal Area (FATA) of Pakistan. Morphological (n = 104) and molecular (n = 54) characterization of the ticks revealed the presence of six ixodid ticks: Rhipicephalus (Rh.) haemaphysaloides, Rh. microplus, Rh. turanicus, Haemaphysalis (Hs.) punctata, Hs. sulcata and Hyalomma anatolicum. Phylogenetic analyses of nucleotide sequence data for two mitochondrial (16S and cytochrome c oxidase 1) and one nuclear (second internal transcribed spacer) DNA regions provided strong support for the grouping of the six tick species identified in this study. Microfluidic real-time PCR, employing multiple pre-validated nuclear and mitochondrial genetic markers, detected 11 potential pathogens and endosymbionts in 72.2% of the ticks (n = 54) tested. Rickettsia (R.) massiliae was the most common pathogen found (42.6% of ticks) followed by Theileria spp. (33.3%), Anaplasma (A.) ovis and R. slovaca (25.9% each). Anaplasma centrale, A. marginale, Ehrlichia spp., R. aeschlimannii, R. conorii and endosymbionts (Francisella- and Coxiella-like) were detected at much lower rates (1.9-22.2%) in ticks. Ticks from goats (83.9%) carried significantly higher microorganisms than those from sheep (56.5%). This study demonstrates that ticks of small ruminants from the FATA are carrying multiple microorganisms of veterinary and medical health significance and provides the basis for future investigations of ticks and tick-borne diseases of animals and humans in this and neighboring regions.

The potential of Allium sativum and Cannabis sativa extracts for anti-tick activities against Rhipicephalus (Boophilus) microplus.

The efficacy of Allium sativum and Cannabis sativa against Rhipicephalus microplus ticks was evaluated using the adult immersion and the larval packet test. In addition, an in silico approach was utilized by performing a docking study in order to identify the active ingredients from both plants. Results showed a comparatively high lethal effect of A. sativum and C. sativa on egg laying (index of egg laying = 0.26 and 0.24, respectively), egg hatching (33.5 and 37.1, respectively), and total larval mortality (100%, both), at 40 mg/mL. When applied to cattle which had been inoculated with larvae ticks, it was observed that a 45% solution of both herbal extracts significantly reduced the number of ticks by 96 h post treatment. We analyzed in silico 27 known active molecules from both plants and identified in the PubChem database to explore the hypothesis that the effect found on ticks was based on inhibition of acetylcholinesterase (AChE). Vitamin E and cannabidiol are the most potent AChE inhibitors with docking scores of -15.85 and -14.38, respectively. Based on these findings, we conclude that A. sativum and C. sativa may potentially be used for the control of R. microplus, and should be further investigated as a potential supplement to or replacement of synthetic acaricides.

Development of herbal formulation of medicinal plants and determination of its antihyperuricemic potential in vitro and in vivo rat's model.

Hyperuricemia is a common metabolic disorder and several herbal formulations are being used for its treatment. The study aimed to develop herbal formulation (Urinil B) and find its hypouricemic effects in vitro and in vivo. Urinil B was prepared by taking Trachyspermum ammi, Piper nigrum and Berberis vulgaris equally. In vitro Dissolution test and xanthine oxidase inhibition assay was performed for checking capsule absorbance and IC50 calculation respectively. For in vivo experimentation, the study comprised of 14 groups of rats (n=6). Results showed that significant xanthine oxidase inhibition was shown by herbal formulation with IC50 of 586±1.5µg/mL. Oral administration of Urinil B 250, 500 and 1000 mg/kg decreased serum and liver uric acid levels of hyperuricemic rats in dose and time dependent manner. 3 day and seven day administration of Urinil B reduced serum and liver uric acid level more significantly as compared to one day administration. However, allopurinol normalized serum and liver uric acid levels in all study groups. The present study indicated marked hypouricemic effects of Urinil B in hyperuricemia induced by potassium oxonate in rats. However, due to caveat of small sample size in this study, clear conclusion regarding hypouricemic potential of Urinil B can't be made.

Molecular epidemiology and associated risk factors of Anaplasma marginale and Theileria annulata in cattle from North-western Pakistan.

Ticks and tick-borne pathogens are one of the major threats to livestock production worldwide. The aim of present study is to specify the molecular epidemiology and its associated risk factors of two well-distributed tick-borne pathogens Anaplasma marginale and Theileria annulata in cattle population from North-western Pakistan. Blood samples were Collected from 60 (32.6 %) local breed cattle, 101 (54.9 %) from crossbreed, and 24 (13.0 %) from exotic breed with total of 184 blood samples. Species-specific PCR assays were performed to detect the presence of A. marginale and T. annulata based on 16S rRNA and 18S rRNA genetic makers respectively. PCR results showed that A. marginale was 16.3 % prevalent and T. annulata was 29.9 % prevalent in the study area with a total prevalence rate of 46.2 % (85/184) of the tested blood samples. District wise analysis revealed that both pathogens were highly prevalent in district Dir Upper (13.6 %) and least prevalent in district Chitral (10.3 %). Univariable analysis of risk factors showed that only breed and acaricidal treatment were significant determinants (P < 0.05) for A. marginale infection, however, in case of T. annulata infection; breed, age, gender, grazing practice, and acaricidal treatment were potential determinants (P < 0.05). Multivariable analysis specified that breed and acaricidal treatment were considered as significant risk factors for A. marginale infection (P < 0.05) whereas acaricidal treatment was found to be a significant determinant for T. annulata infection (P < 0.05). Phylogenetic analysis indicated that A. marginale 16S rRNA and T. annulata 18S rRNA isolates showed similarities and shared phylogeny with same isolates reported from Asia. This is the first molecular report on the epidemiology and risk factors analysis of A. marginale and T. annulata infections in cattle population from NW Pakistan. Further large scale study is required to investigate molecular, epidemiological and genotypic aspects as well as potential risk factors analysis from the country to facilitate designing strategies to control tick-borne pathogen and reduce losses to cattle industry.

An alternative approach to treat obesity with leptogenic polyherbal formulation obesecure: A randomized clinical trial study.

Obesity is a common disease of developing countries, including Pakistan. Obesity is a risk factor for many diseases which can be life threatening or making the person unable to perform daily routine work. In the current study, clinical trials were designed to evaluate the effects of medical intervention by comparing the effects of placebo control drug "Plasicure" with the herbal medicinal formulation "Obesecure". The test drug formulation was designed on the basis of the screening study for Leptogenic drugs. To evaluate the safety of the test drug, the toxicity index and the safety profile of test formulation was assessed on animal models. The drug was found safe for further clinical study. Randomized Controlled Clinical Trials were conducted. The statistical analysis was carried out by the application of Two-Way Repeated Analysis of Variance test. The clinical findings of randomized controlled trial revealed that the test drug was Leptogenic and effective in weight reduction as compared to control drug Plasicure therapy as the p-value deduced was 0.001 in leptin level and 0.000 in case of BMI after the conduction of Two-Way Repeated Analysis of Variance test. Hence it is concluded that obscure therapy is more significant than control drug Plasicure therapy in the management and treatment of obesity.

Prevalence and risk factors associated with prenatal depression among pregnant women in Faisalabad, Pakistan.

To determine the prevalence of prenatal depression and explore its association with socio-demographic risk factors, stressful life events and women autonomy/decision-making. The recognition of these problems would play a substantial role in the strategic development of interventions for those Pakistani women who have been suffering from prenatal depression. Analytical cross-sectional study conducted in Allied hospital Faisalabad during a period of 18 months (1st Jan 2019 to 30 June 2020). Data was collected by using simple random sampling technique. EPDS was used to measure prenatal depression. A modified kuppuswamy scale, Life Events and Difficulties Schedule (LEDS) and women autonomy/decision making scale was used to assess the risk factors. A total of 380 women were included in the study. The mean age (in years) ± SD was 29.37±5.27. According to cut-off score >10 on Edinburg postnatal depression scale the prevalence of depression was found as 51.6%. The findings revealed that women education, socio-economic class, BMI, pregnancy duration, history of miscarriage, contraception, stressful life events and women autonomy had significant association with prenatal depression (P<0.05). Socio-demographic factors, stressful life events in previous life and women autonomy are significant predictors of prenatal depression that need to be addressed in order to sustain safe motherhood.

Quorum Sensing Interfering Strategies and Their Implications in the Management of Biofilm-Associated Bacterial Infections

Abstract The bacterial species employ various types of molecular communication systems recognized as quorum sensing for the synchronization of differential gene expression to regulate virulence traits and biofilm formation. A variety of quorum sensing inhibitors; molecules that interfere with quorum sensing among bacteria have been examined which can block the action of autoinducers. Moreover, the studies have scrutinized various enzymes for their quorum quenching activity resulting in the degradation of signaling molecules or blocking of gene expression. So far, the studies have found that these approaches are not only capable to reduce the pathogenicity and biofilm formation but also resulted in increased bacterial susceptibility to antibiotics and bacteriophages. The effectiveness of these strategies has been validated in different animal models and it seems that these practices will be transformed in near future to develop the medical devices including catheters, implants, and dressings for the prevention of bacterial infections. Although many of these approaches are still in the research stage, the increasing library of quorum quenching molecules and enzymes will open innovative perspectives for the development of antibacterial approaches which will extend the therapeutic arsenal against the pathogenic bacterial species.

Genetic diversity, piroplasms and trypanosomes in Rhipicephalus microplus and Hyalomma anatolicum collected from cattle in northern Pakistan.

The two most important tick species in Pakistan are Rhipicephalus microplus and Hyalomma anatolicum. When associated with cattle, these have one or three host life cycles, respectively, with potential implications for their population genetics and for their vector role in the transmission of pathogens. To compare the two tick species in this context with molecular-phylogenetic methods, during the present study 123 ticks were collected from cattle in northern Pakistan. Two mitochondrial markers of 36 ticks were molecularly analyzed. All 11 R. microplus specimens had identical cox1 haplotypes, whereas the 25 H. anatolicum specimens had nine cox1 haplotypes. The latter belonged to two distinct phylogenetic lineages with high support. However, in the 16S rRNA gene these differences were less evident. Among the 113 ticks molecularly analyzed for tick-borne protozoa, the sequence of Babesia occultans was successfully amplified from two specimens of H. anatolicum. Theileria annulata was present in both R. microplus (10.4%) and H. anatolicum (27.3%), with significantly higher prevalence rate in the latter species. Only one tick, a H. anatolicum female, was positive in the PCR detecting Trypanosoma spp. Sequencing revealed the presence of a new genotype, with the closest phylogenetic relationship to stercorarian trypanosomes (in particular, to a tick-associated Trypanosoma sp. from Japan). In conclusion, the above differences between R. microplus and H. anatolicum may be partly related to their life cycles involving one host or three hosts, respectively. Among the others, host switching (reducing chances of inbreeding) and shorter periods spent on-host (reducing gene flow between cattle herds) are supposed to be important drivers of cox1 gene diversification in case of H. anatolicum as a three host tick species. These results highlight the importance of studying differences in intraspecific genetic diversity and piroplasm burdens between one host and three host ticks in the local scale. In addition, a Trypanosoma sp. molecularly identified in H. anatolicum is reported here for the first time from South Asia, deserving further evaluation concerning its host and vector species.

Acaricidal efficacy of Calotropis procera (Asclepiadaceae) and Taraxacum officinale (Asteraceae) against Rhipicephalus microplus from Mardan, Pakistan.

Medicinal plants are used by traditional folk healers, modern physicians, and veterinarians as an alternative to conventional drugs to treat a wide range of disorders including parasitic diseases. Some compounds from these plants have been shown to have acaricidal activity and repel arthropods. The cattle tick Rhipicephalus microplus is one of the most destructive pests to the livestock industry in tropical and subtropical parts of the world. The potential to develop herbal acaricides to control R. microplus infestations is critical in maintaining cattle herd productivity, reducing economic losses, and curtailing the overuse of synthetic chemical acaricides. Calotropis procera, the apple of Sodom, and Taraxacum officinale, the common dandelion, were evaluated for acaricidal activity against R. microplus larvae and adults in vitro. Both plant species tested are common indigenous species of Pakistan where R. microplus infestations are widespread across livestock species including cattle, sheep, and goats. Whole-plant extracts derived from C. procera and T. officinale significantly reduced the index of egg laying (P < 0.01) and increased the percent inhibition of oviposition of adult female ticks at a concentration of 40 mg/mL when assessed by the adult immersion test (AIT). Calotropis procera and T. officinale treatments at the same concentration also resulted in larval mortality of 96.0% ± 0.57 and 96.7% ± 0.88, respectively, as measured using the larval packet test (LPT). An increasing range of extract concentrations was tested to determine the LD50 and LD90 for C. procera, 3.21 and 21.15 mg/mL, respectively, and T. officinale, 4.04 and 18.92 mg/mL, respectively. These results indicate that further studies are warranted to determine the relative contribution of individual phytochemicals from whole-plant extracts on acaricidal activity. This information will guide the design of further acaricidal efficacy tests using livestock infested with R. microplus.

Quality of life before and after total knee arthroplasty in clinical settings across Lahore, Pakistan.

Knee osteoarthritis is a common disabling condition that affects a larger number of populations aged above 40 years and this habitually required knee arthroplasty to reinstate leg function, to improve the pain as well as to improve stability. The main objective of this study was to determine the health-related quality of lifestyle before and after total knee arthroplasty. This study was a cross-sectional observational study conducted at Shaukat Khanum Memorial Cancer Hospital & Research Centre, Horizon Hospital and Mayo Hospital, Lahore-Pakistan. In this study, 100 subjects were selected who were scheduled for Total Knee Replacement (TKR) due to advanced stages of knee osteoarthritis. Informed consent was taken from all selected patients. Selected subjects meeting inclusion and exclusion criteria were enrolled for this study. Subjects with any red flag signs or systemic illness were excluded. A baseline measurement was taken by using visual-analog scale (VAS) for pain, the Short Form 36 (SF-36) for health-related quality of life and The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) for disability. Measurements using these scales were repeated after 1, 3 and 6 months after knee arthroplasty. Analysis of variance (ANOVA) test was used to determine statistically significant changes in VAS, SF 36 and WOMAC Score across 06 months. A significant (p<0.05) improvement in health-related quality of life measured of two subscales of SF 36 (Physical Health Component and Mental Health Component) when before surgery readings were compared with that of after 06 months of surgery. Similarly, there was also significant (p<0.05) reduction in pain measured on VAS as well as reduced WOMAC score before and after 06 months of surgery. It could be concluded that TKA results in the improvementof the quality of life and reduced pain following total knee arthroplasty.