Genetic Diversity and Structure of Persian Walnut (Juglans regia L.) in Pakistan: Implications for Conservation.

Persian (Common) walnut (Juglans regia L.) is a famous fruit tree species valued for its nutritious nuts and high-quality wood. Although walnut is widely distributed and plays an important role in the economy and culture of Pakistan, the genetic diversity and structure of its populations in the country remains poorly understood. Therefore, using 31 nuclear microsatellites, we assessed the genetic diversity and population structure of 12 walnut populations sampled across Pakistan. We also implemented the geostatistical IDW technique in ArcGIS to reveal "hotspots" of genetic diversity. Generally, the studied populations registered relatively low indices of genetic diversity (NA = 3.839, HO = 0.558, UHE = 0.580), and eight populations had positive inbreeding coefficient (FIS) values. Low among-population differentiation was indicated by AMOVA, pairwise FST and DC. STRUCTURE, PCoA and neighbor joining (NJ) analysis revealed a general lack of clear clustering in the populations except that one population in Upper Dir was clearly genetically distinct from the rest. Furthermore, the Mantel test showed no correlation between the geographic and genetic distance (r = 0.14, p = 0.22), while barrier analysis suggested three statistically significant genetic barriers. Finally, the spatial interpolation results indicated that populations in Ziarat, Kashmir, Dir, Swat, Chitral, and upper Dir had high intrapopulation genetic diversity, suggesting the need to conserve populations in those areas. The results from this study will be important for future breeding improvement and conservation of walnuts in Pakistan.

Superporous acrylic acid and HPMC hydrogels of mefenamic acid: Formulation, characterization and optimization by central composite design.

The purpose of the study was to devise the superporous hydrogels (SPHs) of mefenamic acid (MA) to acquire the sustained action of the MA in the body. The superporous hydrogels of mefenamic acid were formulated by employing the gas blowing method. The central composite rotatable design (CCRD) was applied to optimize the effect of independent formulation factors like acrylic acid (AC), HPMC and glycerol (GLY) over dependent variables like porosity, viscosity, drug content and swelling ratio of superporous hydrogels in water, phosphate buffer (pH 6.8) and in 0.1N HCl (pH 1.2). A number of characteristics such as void fraction, surface morphology by Scanning electron microscopy (SEM) and in vitro drug release study were governed along with physico-chemical analysis by Fourier transform infrared spectroscopy (FTIR), Differential scanning calorimetry (DSC) and appraised statistically by employing the ANOVA. The comparative analgesic activity of optimized superporous hydrogel formulation SPH17 was also analyzed by using tail flick method. The Fourier transform infrared spectroscopy and Differential scanning calorimetry studies approved the physical compatibility between the polymers and the drug. The Scanning electron microscopy study specified micrographic insight about the structure of formed formulations comprising presence of pores, fibers and drug-hole aggregates. The superporous hydrogels were detected to be low dense as they expressed density lower than 0.75 g/cc. The decrease in concentration of the polymers and cross linker contributed towards the increase in the void fraction of the superporous hydrogel formulations. The optimized formulation SPH 17 exhibited a highly sustained release of MA for up to 10 h in the both 0.1 N HCl and phosphate buffer (66.6%) media. The non-fickian release of drug revealed the coupling of the diffusion and polymer relaxation mechanism of the drug release from the formulations. The obtained outcomes suggested that analgesic effect of SPH 17 was significantly (p < 0.05) higher than that of simple suspension of mefenamic acid and total analgesic effect duration for superporous hydrogel was also doubled as compared to the duration of analgesic effect produced by drug suspension. The successfully formulated SPH with HPMC K100M as a gelling agent had sustained the action of the mefenamic acid (MF) by improving its poor solubility and permeability. The introduction of inter-penetrating polymeric network (acrylic acid) using glycerol as a cross linker impart increased residence time to superporous hydrogels which ultimately enhanced the feasibility of using superporous hydrogel as oral sustained release devices particularly for gastric retention.

Optimization of metronidazole SR buccal tablet for gingivitis using genetic algorithm.

Gingivitis is a condition that needs sustained concentration of antibiotic locally over extended period of time. The current study aimed to formulate and evaluate the sustained and localized release of metronidazole (MTZ) as mucoadhesive buccal tablet containing hydroxypropylmethyl cellulose (HPMC), Carbopol 940® (CP), carboxymethylcellulose (CMC) and ethyl cellulose (EC) as mucoadhesive polymers. Tablets were directly compressed with proportions of polymeric blends (F1-F16). The results indicated that weight variation (249±2.10mg) and friability (0.21%) were within USP compendial limits. Maximum mucoadhesive strength and time were depicted by F1 and F14 which were 28.47g and 12hr respectively. Formulations, except F4, were within physiological pH limit. Maximum swellability index (261.9%) was exhibited by F16, at 8 hr, containing highest concentration of CP, HPMC and additional CMC. For in vitro release, the pre-set 8 hr complete release were shown by formulations, F15 and F16 which were 100% and 97%, respectively. Genetic algorithm was applied on the attributes to optimize polymeric response in accordance with desirability. The software predicted composition (F17) was tested which revealed that physical characteristics were in accordance with the compendial standards. The release kinetics, evaluated through DDsolver⌖, suggested that release of MTZ followed non-Fickian diffusion type in Korsmeyer-Peppas model. Therefore, MTZ, if delivered as mucoadhesive buccal formulation (F17) containing amounts (mg) of CP (16.4), HPMC (78.7), CMC (8.3) and EC (10.5) will simulate satisfactory release i.e. 96% at 8 hr in simulated buccal fluid.

Frequency and genotype distribution of high risk human papillomavirus in esophageal squamous cell carcinoma.

INTRODUCTION: Esophageal squamous cell carcinoma (ESCC) is a dismal disease exhibiting striking geographical differences in its incidence. It is multifactorial in origin. Among infectious agents, human papillomavirus (HPV) was introduced as a possible causative agent in the development of ESCC in 1982. Subsequent studies using various methods have confirmed the presence of HPV in ESCC. We aimed to determine the frequency of HPV in ESCC in northwest Pakistan which is part of high risk belt for this disease. METHODOLOGY: This study was conducted on two hundred and forty-three (243) diagnosed cases of ESSC at two tertiary care hospitals of Peshawar, Pakistan, from 2011 to 2016. DNA was extracted from all specimens. Polymerase chain reaction (PCR) was used to check the quality of DNA using ß-globin primers and frequency and genotypes of HPV using HPV general primers and type-specific primers respectively. HPV and its genotypes were confirmed through the sequencing of a few selected cases. RESULTS: Two hundred and three (203) tissue specimens had adequate DNA and were further analyzed. HPV positivity with general primers alone was 15.7% (32/203). Using HPV general primers and type-specific primers (HPV 16 or HPV 18), the overall positivity of HPV was 31% (63/203). For type-specific primers, frequency of HPV types 16 and 18 was 20.19% (41) and 7.8% (16) respectively where 6 cases were positive for both HPV 16 and 18. CONCLUSIONS: The overall high prevalence of HPV indicates it as a possible risk factor for ESSC.

FOREIGN BODY IN URINARY BLADDER: AN UNUSUAL PRESENTATION.

Urinary bladder functions for the storage and expulsion of urine. Foreign bodies in male urinary bladder are rare due to lengthy urethra, especially when they are rounded in shape and difficult to be introduced into urinary bladder. This is a case report of a middle age male with thin body habitus and slightly disturbed psychiatric status. He presented with both irritative and obstructive lower urinary tract symptoms (LUTS). He was investigated and found to have an intact bangle in the urinary bladder. The patient was planned for surgery, and the foreign body was removed endoscopically with the help of cystoscope. Later on the patient was referred to a psychiatrist for psychological management.

Secondary defense antioxidant status of vitamin C, vitamin E and GSH in malaria, caused by plasmodium Falciparum and plasmodium Vivax.

Present investigations focused on the antioxidant defense in malaria caused by plasmodium Falciparum and plasmodium Vivax the mean±SEM values of Vitamin C, Vitamin E and GSH very highly significantly decreased as compared with normal individuals in both malaria species which cause malaria disease. The antioxidant levels in female were decreased very significantly due to decreased levels of antioxidant as compared with the male patients. The results are shown as mean±SME, the antioxidant levels in malarial patients was compared with normal individuals, in both genders. The antioxidant levels of vitamin C, Vitamin E and glutathione decreased in malaria caused by both species, a much greater decrease in patients infected by Plasmodium Vivax. P>001 was considered significant. The decrease of antioxidant levels was higher in female patients as compared with male patients. Antioxidant supplements like Vitamin C, Vitamin E and GHS may be used with anti malarial therapy,as a preventive measure because malaria affects the secondary antioxidant defense system of the body.

Chemical weed management in wheat at higher altitudes.

To evaluate the effect of different herbicides for controlling weeds in wheat (variety Fakhr-i-Sarhad),at higher attitude, an experiment was conducted at Agriculture Research Station, Chitral during Rabi season 2003-04, using Randomized Complete Block Design, keeping four replications. The experiment, sown in November comprised of eight treatments, viz; seven herbicides and a weedy check. Each treatment consisted of 5 rows each 30 cm apart and 5 m long thus giving a total size of 5 m x 1.5 m. The herbicides used included; terbutryn + triasulfuron at 0.16 kg, 2,4-D at 0.7 kg, fenoxaprop-P-ethyl at 0.93 kg, clodinafop at 0.05 kg, bromoxynil + MCPA at 0.49 kg, carfentrazon-ethyl at 0.02 kg and isoproturon at 1.0 kg a.i ha(-1). The data were recorded on weed kill efficiency (%), fresh weed biomass (kg ha(-1)), plant height (cm), spike length (cm), number of tillers m(-2), number of grains spike(-1), thousand grains weight (g), biological yield (kg ha(-1)), grain yield (kg ha(-1)) and harvest index (%). The data recorded on weed kill efficiency, weed biomass (kg ha1), grains yield (kg ha(-1)) and harvest index (%) were significantly affected by the different herbicidal treatments. Statistically isoproturon treatment exhibited the best performance, with maximum weed kill efficiency (48.26%) and minimum fresh weed biomass (433.3 kg ha(-1)) as compared to weedy check (6 %) and (1102 kg ha(-1)), respectively. Similarly, the spike length (8.34 cm), number of tillers (427 m(-2)), number of grains spike(-1) (38.0), thousand grains weight (39.85 g), biological yield (8475 kg ha(-1)), grain yield (2530 kg ha(-1)) and harvest index (31.3%) were the highest in isoproturon treatments as compared to weedy check having (7.64 cm), (356 m(-2)), (34.1), (37.12 g), (6858 kg), (1913 kg ha(-1)) and (27%), respectively.

Application of alvarado scoring system in diagnosis of acute appendicitis.

BACKGROUND: In conditions like acute appendicitis it is impractical to have definitive diagnosis by gold standard test (histopathology) before surgery, we would like a simple test like Alvarado scoring system which depends on the presence and absence of certain variables and which provides an accurate guide to whether or not the patient has the condition. This study was conducted to evaluate Alvarado scoring system for the diagnosis of acute appendicitis in our set up. METHODS: 100 consecutive patients with suspected acute appendicitis admitted in Surgical B unit, Khyber Teaching Hospital Peshawar, during the period from July to December 2003 were included in the study. They were given specific scores according to the variables of Alvarado scoring system and then divided into 3 groups. Group 1 patients (score 7 or more) underwent surgery, group 2 patients (score 5-6) were admitted for observation and group 3 patients (score 4 or less) were discharged home. Patients from group 2 with increased symptom intensity (score 7 or more) in reevaluation underwent surgery. Diagnosis was confirmed by histopathological examination. Reliability of scoring system was assessed by calculating negative appendicectomy rate and positive predictive value. RESULTS: Out of a total 100 patients 64 patients underwent surgery and appendicitis was confirmed in 54 cases, thus giving negative appendicectomy frequency of 15.6% (male 12%, female 17.9%). Perforation rate was 7.8%. Positive predictive value was 84.3% (males 88%, females 82.1%). CONCLUSION: This scoring system is easy, simple and cheap complementary aid for supporting the diagnosis of acute appendicitis especially for junior surgeons.

Mastoiditis in children.

Fifty-two children with mastoiditis were treated at the Federal Government Services Hospital over a period of 15 years (from 1986 to 2000). The salient feature in these cases was the presence of a post-auricular swelling and mastoid abscess. Extensive cholesteatoma was seen in 80.7 per cent of cases and 19.3 per cent had granulations in the mastoid cavity. Significantly, all patients had a history of chronic ear discharge prior to abscess formation. These children belonged to poor communities, were malnourished and attended hospital in a state of emergency. The presentation and course of the disease confirms the aggressive and potentially serious nature of the pathology requiring early medical attention in order to avert impending complications. It appears that in our environment acute mastoiditis with abscess formation develops from chronic otitis media and the cholesteatoma. The surgical procedures conducted in these cases were radical and modified radical mastoidectomy.

Middle-ear reconstruction: a review of 150 cases.

This study is based on a retrospective analysis of 150 cases of tympanoplasty and ossicular chain reconstruction as a one-stage procedure. It was conducted at the Federal Government Services Hospital, Islamabad from 1983 to 1999. Temporalis fascia was used for myringoplasty and a sculptured autologous incus to bridge the malleus to stapes head and malleus to footplate gap. These interpositions have produced stable ossicular assemblies and provided satisfactory post-operative hearing gains. One hundred and twenty-six (84 per cent) patients achieved a healed tympanic membrane three months post-operatively. One year post-operatively, 121 (81 per cent) patients had an intact tympanic membrane and at five years 119 (79 per cent) had an intact tympanic membrane. The success rate was based on an intact tympanic membrane and an air-bone gap of less than 30 dB one-year post-operation. Accordingly, the overall success rate was 80 per cent (120 cases). Hearing gains in cases with an absent stapes arch were poorer than in cases with an intact arch. Early (one to three months) post-operative complications included tympanic membrane perforation, lateralization of the graft and persistence of infection. Late (one to five years) complications included the appearance of an anterior perforation and retraction and thinning of the tympanic membrane.