High frequency and founder effect of the CYP3A4*20 loss-of-function allele in the Spanish population classifies CYP3A4 as a polymorphic enzyme.

Cytochrome P450 3A4 (CYP3A4) is a key drug-metabolizing enzyme. Loss-of-function variants have been reported as rare events, and the first demonstration of a CYP3A4 protein lacking functional activity is caused by CYP3A4*20 allele. Here we characterized the world distribution and origin of CYP3A4*20 mutation. CYP3A4*20 was determined in more than 4000 individuals representing different populations, and haplotype analysis was performed using CYP3A polymorphisms and microsatellite markers. CYP3A4*20 allele was present in 1.2% of the Spanish population (up to 3.8% in specific regions), and all CYP3A4*20 carriers had a common haplotype. This is compatible with a Spanish founder effect and classifies CYP3A4 as a polymorphic enzyme. This constitutes the first description of a CYP3A4 loss-of-function variant with high frequency in a population. CYP3A4*20 results together with the key role of CYP3A4 in drug metabolism support screening for rare CYP3A4 functional alleles among subjects with adverse drug events in certain populations.

Theoretical investigation of thermoelectric properties of methyl blue-based molecular junctions.

Thermoelectric properties of a family of methyl blue-based molecular junctions were theoretically studied using a combination of density functional theory (DFT) methods, and quantum transport theory (QTT). Employing different numbers of amino groups not only proves itself as a powerful strategy for controlling the transport behaviour and lifting the transmission coefficient T(E) from 1.91 × 10-5 to 7.45 × 10-5 with increasing the amino groups from zero to four, but also it enhances the thermoelectric properties of these molecules, since it increases the Seebeck coefficient (S) from 106.8 to 202.4 µV K-1 and the electronic figure of merit (Z el T) has been raised from 0.15 to 0.35, making these molecules promising candidates for thermoelectric applications.

Anxiolytic potential of resveratrol and rufinamide combination by modulating GABA-ergic transmission: insights from experiments, molecular docking and dynamics simulations.

Resveratrol is a polyphenolic phytocompound known to possess anxiolytic-like effects but its impact on central gammaaminobutyric acid (GABA) modulation has never been explored. The purpose of this study was to analyze the anxiolytic-like effects of resveratrol alone and in combination with rufinamide, an antiepileptic drug which has never been studied for its anxiolytic potential. The BALB/c mice were tested in a battery of behavior testing after administration of resveratrol (50 mg/kg) and rufinamide (50 mg/kg) alone and in combination. Moreover, molecular docking studies were also carried out to understand the interaction of resveratrol and rufinamide with GABA aminotransferase, GABA receptor and GABA-A transporter type 1. Resveratrol alone exerted notable anxiolytic-like effects and improved outcomes in few experiments but rufinamide alone did not yield any beneficial outcomes. However, the animal co-administered with resveratrol and rufinamide behaved exceptionally well (p<0.05) and preferred open, illuminated and exposed areas of open field, light/dark and elevated plus maze. Further, these animals showed reduced anxiety towards anxiogenic stimuli i.e. holes and marbles in hole board and marble bury tests, respectively. Resveratrol and rufinamide showed moderate to strong binding affinities with GABA proteins, indicating the potential to treat anxiety-like neurological disorders. Moreover, resveratrol and rufinamide were analyzed using molecular docking to determine their interaction with GABA receptors, transporters, and transaminase. The results suggest that their anxiolytic-like effects may be due to inhibiting GABA reuptake transporter 1 protein, leading to increased synaptic levels of GABA neurotransmitter, as seen in stable molecular dynamics results with the 7SK2 GABA transporter protein.

Prevalence and risk factors of Syphilis among blood donors of Punjab, Pakistan.

Syphilis is a sexually transmitted disease and its actual prevalence among Pakistani blood donors is currently unknown. A cross sectional study was conducted at different district healthcare hospitals of Punjab, Pakistan with an aim to evaluate the prevalence and risk factors associated with syphilis in blood donors using immunochromatographic test (ICT) and enzyme linked immunosorbent assay (ELISA). A total (n=1200) blood samples were collected from donors aged 18-65 years. All the information regarding personal data, demographic data and risk factors was collected via structured questionnaire. On the basis of ICT and ELISA, the overall prevalence of syphilis was 3.91% among blood donors. The demographic factors positively linked with syphilis were age (P= 0.000; Odds ratio, OR= 7.18; 95% confidence interval CI= 2.816-18.295) and education status (P= 0.000; Odds ratio, OR= 12.33; 95% confidence interval CI= 3.469-43.849) of donors. Similarly among the risk factors analyzed, marital status (P= 0.012; Odds ratio OR= 2.251; 95% confidence interval CI= 1.206- 4.202) and blood transfusion history (P= 0.030; Odds ratio OR= 1.981; 95% confidence interval CI= 1.083-3.623) were also strongly associated with syphilis. We emphasized the importance of promoting preventive measures for syphilis. The syphilis diagnosis should not be based on a single test. The present study indicates that higher prevalence is alarming for blood donors in Pakistan. Stringent donor screening is highly recommended to ensure maximum safe blood transfusion.

Neuropharmacological investigation, ultra-high performance liquid chromatography analysis, and in silico studies of Phyla nodiflora.

The increasing burden of neurological disorders is becoming a worldwide health challenge and researchers are continuously struggling to cure them by utilizing the miraculous medicinal properties of plants. The crude methanolic extract of whole herb of Phyla nodiflora (Pn.Cr) was subjected to phytochemical, antioxidant and neuropharmacological assessment. The Pn.Cr was initially exposed to the in vitro examination for phytocomposition through ultra-high performance liquid chromatography (UHPLC). The Sprague Dawley rats were chronically administered with various doses (100, 200 and 300 mg/kg) of Pn.Cr for one month with subsequent exposure to neurobehavioral and biochemical experimentation. The Pn.Cr exhibited a dose-dependent anxiolytic effect (P < 0.05 in comparison to control) as rats preferred central, illuminated and open arm zones in open field (OFT), light/dark (L/D) and elevated plus maze (EPM) tests. Likewise, scopolamine-induced amnesia was noticeably reversed with P < 0.05 by Pn.Cr as animals showed improved spontaneous alternation, discrimination index and shorter escape latencies in Y-maze, novel object recognition (NOR) and Morris water maze (MWM) tests. Subsequently, in vivo enzymatic assays depicted the reduced acetylcholinesterase and malondialdehyde levels. The levels of oxidative stress combating enzymes (glutathione peroxidase and superoxide dismutase) were increased in a dose-dependent style. The UHPLC detected 22 phytocompounds were further investigated in silico studied to predict the interaction of blood-brain barrier (BBB) crossing phytocompounds with human acetylcholinesterase. The four BBB crossing phytocompounds belonging to flavonoids, chalcones and alkaloids showed possible interaction with the target enzyme. We found that the phytocompounds owned by Pn.Cr might be playing multiple roles in modulation of different pathways to hinder the pathophysiology of neurological disorders including anxiety and Alzheimer's disease.

Preparation of TiO2/WO3/C/N Composite Nanofibers by Electrospinning Using Precursors Soluble in Water and Their Photocatalytic Activity in Visible Light.

Extending the absorption range of TiO2 nanofibers to visible light is a great improvement of the photocatalytic property of TiO2. In this study, TiO2/WO3/C/N nanofibers were prepared by electrospinning using precursors soluble in water then annealing in argon. Titanium(IV) bis(ammonium lactato)dihydroxide (TiBALDH) and ammonium metatungstate (AMT) were used as the precursor for TiO2 and WO3 respectively. Different volume ratios of the precursors were added to a solution of PVP before electrospinning. The fibers were studied by XPS, SEM-EDX, TEM, FTIR, XRD, Raman spectroscopy and UV-VIS diffuse reflectance spectroscopy (DRS). The photocatalytic degradation of methylene blue by the fibers in visible light was investigated. The fibers had anatase TiO2 and monoclinic WO3. Based on UV-VIS DRS and Kubelka-Munk function the fibers could absorb visible light. Moreover, 100% TiBALDH had an indirect band gap of 2.9 eV, and the band gap decreased with increase in AMT, i.e., for 0% TiBALDH, band gap was 2.4 eV. The fibers degraded methylene blue dye in visible light, and 90% TiBALDH had the highest photocatalytic activity, i.e., it degraded 40% of the dye after 240 min.

Time course evaluation of lacosamide alone and in polypharmacy on behavioral manifestations and oxidative stress in lithium-pilocarpine-induced model.

The lithium-pilocarpine model in rats is commonly used to study the characteristic events of acute status epilepticus (SE), epileptogenesis and temporal lobe epilepsy (TLE). Here we investigated the impact of lacosamide alone and in combination with other drugs (pregabalin, piracetam and scopolamine) on spontaneous recurrent seizures (SRSs) and behavioral parameters during the time frame of 6 weeks after SE. In addition, the level of oxidative stress in the hippocampus was accessed by real-time microdialysis study (8-isoprostanes) and antioxidants enzymes in the homogenate. Results revealed severe behavioral deficits with the control epileptic group and animals displayed hyperexcitability, aggression apprehension and memory insufficiency. Pharmacological manipulation for 6 weeks with lacosamide (L) - 80 mg/kg; in polypharmacy with pregabalin (L/P) - 50/50 mg/kg and piracetam (L/Pi) - 50/140 mg/kg significantly (P < 0.05) ameliorated the anxiety-related behavior (open filed, elevated plus maze, light/dark tests), depression (forced swim test) and improved spatial/reference memory (Morris water maze). There were low incidences of seizures in L, L/P and L/Pi groups revealing disease-modifying effects of employed drugs. Furthermore, the chronic use of scopolamine (L/P/S; 50/50/2 mg/kg) as polypharmacy with the concept of antagonizing the cholinergic inputs in the epileptogenic phase aberrated the behavioral situation further worse. Treatments with L/P and L/Pi significantly attenuated (P < 0.05) the oxidative stress by reducing 8-isoprostanes and malondialdehyde (MDA) levels. Furthermore, superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels in the L/P group were significantly (P < 0.05) improved. Overall, our findings support the use of a combination of drugs (L/P and L/Pi) in lithium-pilocarpine model which remarkably ameliorated SRSs, reduced anxiety-related behaviors, retention of spatial/reference memory and lowered oxidative stress in a time-course evaluation 6 weeks post- SE insult.

Association between ABO blood groups and susceptibility to COVID-19: profile of age and gender in Iraqi patients.

Background: A case-control study was performed to examine age, gender, and ABO blood groups in 1014 Iraqi hospitalized cases with Coronavirus disease 2019 (COVID-19) and 901 blood donors (control group). The infection was molecularly diagnosed by detecting coronavirus RNA in nasal swabs of patients. Results: Mean age was significantly elevated in cases compared to controls (48.2 ± 13.8 vs. 29.9 ± 9.0 year; probability [p] < 0.001). Receiver operating characteristic analysis demonstrated the predictive significance of age in COVID-19 evolution (Area under curve = 0.858; 95% CI: 0.841 - 0.875; p < 0.001). Males outnumbered females in cases (60.4 vs. 39.6%) and controls (56 vs. 44%). Stratification by age group (< 30, 30 - 39, 40 - 49 and ≥ 50 years) revealed that 48.3% of cases clustered in the age group ≥ 50 years. ABO blood group analysis showed that group A was the most common among cases, while group O was the most common among controls (35.5 and 36.7%, respectively). Blood groups A (35.5 vs. 32.7; corrected p [pc] = 0.021), A+AB (46.3 vs. 41.7%; pc = 0.021) and A+B+AB (68.0 vs. 63.3%; pc = 0.007) showed significantly elevated frequencies in cases compared to controls. Logistic regression analysis estimated odds ratios (ORs) of 1.53 (95% confidence interval [CI]: 1.16 - 2.02), 1.48 (95% CI: 1.14 - 1.93) and 1.50 (95% CI: 1.17 - 1.82) for blood groups A, A+AB and A+B+AB, respectively. Blood group frequencies showed no significant differences between age groups of cases or controls. Regarding gender, male cases were marked with increased frequency of group A (39.9 vs. 28.9%) and decreased frequency of group O (25.9 vs. 41.0%) compared to female cases. Independent re-analysis of ABO blood groups in male and female cases demonstrated that group A was increased in male cases compared to male controls (39.9 vs. 33.1%; OR = 1.65; 95% CI: 1.24 - 2.21; pc = 0.006). On the contrary, no significant differences were found between females of cases and controls. Conclusions: The study results indicated that blood group A may be associated with an increased risk of developing COVID-19, particularly in males.

Prevention of beta-thalassemia in a large Pakistani family through cascade testing.

BACKGROUND: We report cascade testing of a large Pakistani family for beta-thalassemia alleles. The family was still practicing consanguineous marriages and was at risk of having more affected births. OBJECTIVE: The objective of this study was to show that identification of disease carriers in families with index cases in order to create awareness about disease and provide genetic counseling would result in reduction of the frequency of beta-thalassemia in Pakistan. METHODS: In this large family with an index case, 27 available living members were tested for beta-thalassemia. Carriers of the disease were detected by measuring hemoglobin indices, and amplification refractory mutation system polymerase chain reaction was used for mutation analysis. Genetic counseling was provided to members of this family. RESULTS: There were already 3 marriages between the carrier members and 1 between a carrier and noncarrier in this large family; 12 (44.4%) members were found to carry the mutant gene, representing a very high carrier rate compared to the 5.4% carrier frequency of beta-thalassemia in the general population of Pakistan. The family was counseled for prevention of affected births. The initially reluctant family gradually became cooperative and seriously attended the genetic counseling sessions. CONCLUSION: Cascade testing is more practical than general population screening in a country with limited health facilities where consanguineous marriages are practiced. This report emphasizes the need of extensive testing within families with index cases to identify the carriers of beta-thalassemia in order to reduce disease occurrence through awareness and genetic counseling.

An in vivo and in vitro potential of Indian ayurvedic herbal formulation Triphala on experimental gouty arthritis in mice.

In the present study, we have investigated the efficacy of Indian ayurvedic herbal formulation Triphala on monosodium urate crystal-induced inflammation in mice; an experimental model for gouty arthritis and compared it with that of the non-steroidal anti-inflammatory drug, Indomethacin. The anti-arthritic effect of Triphala was evaluated by measuring changes in the paw volume, lysosomal enzyme activities, lipid peroxidation, anti-oxidant status and inflammatory mediator TNF-alpha in control and monosodium urate crystal-induced mice. The levels of beta-glucuronidase and lactate dehydrogenase were also measured in monosodium urate crystal-incubated polymorphonuclear leucocytes (PMNL). Triphala treatment (1 gm/kg/b.w. orally) significantly inhibited the paw volume and the levels of lysosomal enzymes, lipid peroxidation and inflammatory mediator tumour necrosis factor-alpha; however the anti-oxidant status was found to be increased in plasma, liver and spleen of monosodium urate crystal-induced mice when compared to control mice. In addition, beta-glucuronidase and lactate dehydrogenase level were reduced in Triphala (100 microg/ml) treated monosodium urate crystal-incubated polymorphonuclear leucocytes. In conclusion, the results obtained clearly indicated that Triphala exerted a strong anti-inflammatory effect against gouty arthritis.

Protective effect of Withania somnifera root powder in relation to lipid peroxidation, antioxidant status, glycoproteins and bone collagen on adjuvant-induced arthritis in rats.

The present investigation was carried out to evaluate the protective effect of Withania somnifera Linn. Dunal (family-Solanaceae), commonly known as Ashwagandha, on adjuvant-induced arthritic rats. Results were compared with those for Indomethacin, a nonsteroidal anti-inflammatory drug. Arthritis was induced by intradermal injection of complete Freund's adjuvant (0.1 mL) into the right hind paw of Wistar albino rats. Withania somnifera root powder (1000 mg/kg/day) and Indomethacin (3 mg/kg/day) were orally administered for 8 days (from 11th to 18th day) after adjuvant injection. The anti-arthritic effect of W. somnifera root powder was assessed by measuring changes in lipid peroxidation, antioxidant status, and glycoprotein levels in plasma and spleen of arthritic animals. In addition, cartilage degradation was also assessed by estimating bone collagen, and urinary constituents in arthritic animals. Results of the present investigation showed significant increase in the level of lipid peroxides, glycoproteins, and urinary constituents with the depletion of antioxidant status and bone collagen in arthritic animals. These biochemical alterations observed were ameliorated significantly by oral administration of W. somnifera root powder (1000 mg/kg body weight) in arthritic animals. The results of this study clearly indicate that W. somnifera root powder is capable of rectifying the above biochemical changes in adjuvant arthritis.

Micro and macro geographical analysis of Y-chromosome lineages in South Iberia.

In this study, we intend to identify the evolutionary footprints of the South Iberian population focusing on the Berber and Arab influence, which has received little attention in the literature. Analysis of the Y-chromosome variation represents a convenient way to assess the genetic contribution of North African populations to the present-day South Iberian genetic pool and could help to reconstruct other demographic events that could have influenced on that region. A total of 26 Y-SNPs and 17 Y-STRs were genotyped in 144 samples from 26 different districts of South Iberia in order to assess the male genetic composition and the level of substructure of male lineages in this area. To obtain a more comprehensive picture of the genetic structure of the South Iberian region as a whole, our data were compared with published data on neighboring populations. Our analyses allow us to confirm the specific impact of the Arab and Berber expansion and dominion of the Peninsula. Nevertheless, our results suggest that this influence is not bigger in Andalusia than in other Iberian populations.

Immunomodulatory role of Withania somnifera root powder on experimental induced inflammation: An in vivo and in vitro study.

The aqueous suspension of Withania somnifera root powder was investigated for their in vivo and in vitro immunomodulatory properties. W. somnifera showed potent inhibitory activity towards the complement system, mitogen induced lymphocyte proliferation and delayed-type hypersensitivity reaction. Administration of W. somnifera root powder did not have a significant effect on humoral immune response in rats. Our results report immunosuppressive effect of W. somnifera root powder, thus it could be a candidate for developing as an immunosuppressive drug for the inflammatory diseases.

Cancer cervix: An uncommon malignancy in Kashmir, India.

OBJECTIVE: To study the distribution of cancers among females with particular emphasis on cancer cervix in Kashmiri population, which is geographically and socio-culturally distinct from the rest of India. MATERIALS AND METHODS: All patient records were screened from January 1, 2009 to December 31, 2011 at Regional Cancer Centre, Srinagar. Most common cancers among females were recorded and analysis of cancer cervix cases was performed. RESULTS: Female cancers comprised of 40% of total cancers with oesophageal and breast cancer as most common malignancies. Cancer cervix did not figure in top ten cancers and only 45 (0.01%) cases were recorded of the total of 3084 adult female cancers. CONCLUSION: We conclude that due to different socio-cultural and sexual practices, this cancer is highly uncommon in Kashmir and screening or possibly should be directed specifically at only high risk selective subjects.

Fabrication of silicone oral splints for severe burn microstomia in children.

This paper describes a simple technique of fabrication of oral splints (from silicone blocks), which can be utilized in the postoperative period following the release and graft of anterior oral contractures in children. Advantages of the silicone splint when compared to standard acrylic splints are discussed.

Locoregional recurrence after management of carcinoma breast.

OBJECTIVE: To determine the rate of locoregional recurrence, disease-free interval, site and pattern of locoregional recurrence and the significance of different factors for predicting locoregional recurrence in patients with stage II and III carcinoma breast. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Surgical Unit-1, Jinnah Hospital, Lahore from September 2001 to December 2002. PATIENTS AND METHODS: Criteria for including patients in this study was female patients with stage II and III carcinoma breast who presented in our unit from 1995-2002 and underwent surgical (modified radical mastectomy, simple mastectomy with axillary sampling) and non-surgical (chemotherapy, radiotherapy, hormonal therapy) treatment of carcinoma breast. Record of 98 patients was evaluated for rate of locoregional recurrence, disease-free interval, site and pattern of locoregional recurrence and different clinical factors like stage of carcinoma, tumour size, lymph node status and histopathology were assessed for association with locoregional recurrence. RESULTS: After a mean follow-up of 3.5 years, 17 patients developed locoregional recurrence. Mean disease free interval in stage II was 30 months and only 9.5 months in stage III. Lymph node positivity was associated with locoregional recurrence (p-value<0.05). Chest wall was commonest site of recurrence (73%). Single spot recurrence was common in stage II whereas multiple spot recurrence and field change was more common in stage III. Addition of radiotherapy to surgery decreased the locoregional recurrence but chemotherapy had no significant effect on prevention of locoregional recurrence. CONCLUSION: In this series 17.34 % patients developed locoregional recurrence for mean follow-up duration of 3.5 years. Mean disease-free interval was 20.52 months. Lymph node involvement had significant correlation with LRR.

A novel derivative pentacyclic triterpene and omega 3 fatty acid.

Adjuvant induced arthritis (AIA) is accompanied by marked changes in the levels of lysosomal enzymes, glycoproteins and metabolic turnover of collagen. The role of a pentacyclic triterpene and eicosapentaenoic acid (EPA) derivative--Lupeol-EPA (50 mg/kg body weight--orally) was tested in vivo in rats. The increased activities of lysosomal enzymes and glycoproteins associated with decreased collagen in arthritic animals were significantly altered to nearly that of controls. Indomethacin (3 mg/kg body weight) was used as a reference compound. The therapeutic usefulness of Lupeol-EPA derivative in inflammatory conditions is attractive and deserves further work in this direction.

Osseous sporotrichosis.

We report on four patients with osteomyelitis due to Sporothrix schenckii who were successfully treated with potassium iodide.

Dislocations of the elbow in children.

A total of 33 children were treated for acute traumatic dislocation of the elbow between 1994 and 2002; 30 dislocations were posterior and three anterior. Eight children had a pure dislocation and 25 had an associated fracture of the elbow. Two had compound injuries. Two children had injury to the ulnar nerve, one to the radial nerve and one to the median nerve together with injury to the brachial artery. Twenty required open reduction. Complications included pseudarthrosis of the medial epicondyle in one child and loss of flexion and rotation of between 10 degrees and 30 degrees in ten others. Meticulous clinical and radiological assessment is mandatory in children with dislocation of the elbow to exclude associated injuries. The results were excellent to good in 22 patients, fair in ten and poor in one.

Primary subacute haematogenous osteomyelitis in children.

Between 1990 and 1998 we saw 21 children with primary subacute haematogenous osteomyelitis. Pain, swelling and a limp had been present for two to 12 weeks with little functional impairment. Laboratory tests were non-contributory. The lesions were classified radiologically into metaphyseal, diaphyseal, epiphyseal and vertebral. There were 24 sites involved, with most (20) being in the tibia; 17 lesions were in the diaphysis, five in the metaphysis and two in the epiphysis. The diagnosis was confirmed histologically in all cases. Staphylococcus aureus was cultured in six patients. Healing occurred in all patients after treatment with antibiotics for six weeks and radiological improvement was seen after three to six months. Subacute osteomyelitis develops as a result of increased host resistance and decreased bacterial virulence. The radiological features can mimic various benign or malignant bone tumours and non-pyogenic infections. Histological confirmation is necessary to avoid a delay in diagnosis.