Consumption of ß-Caryophyllene Increases the Mating Success of Bactrocera zonata Males (Diptera: Tephritidae).

The peach fruit fly, Bactrocera zonata (Saunders) (Diptera: Tephritidae), is an economically important polyphagous quarantine pest of horticultural crops endemic to South and Southeast Asia. Methyl eugenol (ME), a naturally occurring phenylpropanoid, is a male attractant used to lure and (when mixed with an insecticide) annihilate the males from the wild population, a method of pest control termed the male annihilation technique (MAT). ME is reported to enhance the mating success of sterile males of Bactrocera spp., which is critical for enhancing the effectiveness of the sterile insect technique (SIT). The suppressed response of ME-treated males to ME-baited traps/devices allows the simultaneous application of the MAT and SIT, increasing the efficiency of area-wide integrated pest management (AW-IPM) programs. However, ME treatment in sterile males in SIT facilities is logistically difficult. ß-caryophyllene (BCP) is a widely occurring, safer plant compound and is considered suitable for treating males in SIT facilities. Here, we demonstrate that BCP feeding enhanced B. zonata male mating success to the same extent as ME feeding. Feeding on BCP suppressed the male's subsequent attraction to ME-baited traps, but not to the same degree as feeding on ME. The results are discussed and BCP is suggested as an alternative to ME for the concurrent use of the MAT and SIT.

Polyphenolic characterization and biological assessment of Acacia nilotica (L.) wild. Ex delilie: An In vitro and In vivo appraisal of wound healing potential.

ETHNOPHARMACOLOGICAL RELEVANCE: Acacia nilotica (L.) Wild. Ex Delilie is a shrub with significant ethnomedicinal stature. Therefore, in the undertaken study, its wound healing attributes are determined. AIM OF THE STUDY: The current study provided evidence of the traditional use of A. nilotica species and conferred A. nilotica bark extract as a potent candidate for wound healing agents. MATERIALS & METHODS: A. nilotica leaves extract (ANL-E); A. nilotica bark extract (ANB-E), and A. nilotica stem extract (ANS-E) were prepared using methanol-chloroform (1:1). Phytochemical analysis was performed using gallic acid equivalent (GAE) total phenolic content (TPC), quercetin equivalent (QE) total flavonoid content (TFC) assays and High-performance liquid chromatography (HPLC). In vitro antioxidant potential (free radical scavenging activity (FRSA), total antioxidant capacity (TAC), and ferric reducing antioxidant power (FRAP) assay), antibacterial activity (broth microdilution method) and hemolytic analysis was carried out. Wound healing proficiency of ANB-E was determined by wound excision model followed by estimating hydroxyproline content and endogenous antioxidant markers. RESULTS: Maximum phenolic and flavonoid content were depicted by ANB-E i.e., 50.9 ± 0.34 µg gallic acid equivalent/mg extract and 28.7 ± 0.13 µg quercetin equivalent/mg extract, respectively. HPLC analysis unraveled the presence of a significant amount of catechin in ANL-E, ANB-E and ANS-E (54.66 ± 0.02, 44.9 ± 0.004 and 31.36 ± 0.02 µg/mg extract) respectively. Highest percent free radical scavenging activity, total antioxidant capacity, and ferric reducing action power (i.e., 93.3 ± 0.42 %, 222.10 ± 0.76, and 222.86 ± 0.54 µg ascorbic acid equivalent/mg extract) were exhibited by ANB-E. Maximum antibacterial potential against Staphylococcus aureus was exhibited by ANB-E (MIC 12.5 µg/ml). Two of the extracts i.e., ANL-E and ANB-E were found biocompatible with less than 5 % hemolytic potential. Based upon findings of in vitro analysis, ANB-E (10, 5, and 2.5 % w/w, C1, C2, and C3, respectively) was selected for evaluating its in vivo wound healing potential. Maximum contraction of wound area and fastest epithelization i.e., 98 ± 0.05 % and 11.2 ± 1.00 (day) was exhibited by C1. Maximum hydroxyproline content, glutathione, catalase, and peroxidase were demonstrated by C1 i.e., 15.9 ± 0.52 µg/mg, 9.3 ± 0.17 mmol/mg, 7.2 ± 0.17 and 6.2 ± 0.14 U/mg, respectively. Maximal curbed lipid peroxidation i.e., 0.7 ± 0.15 mmol/mg was also depicted by C1. CONCLUSIONS: In a nutshell, the current investigation endorsed the wound healing potential of ANB-E suggesting it to be an excellent candidate for future studies.

Mechanistic insight into the synergistic antimicrobial potential of Fagonia indica Burm.f. extracts with cefixime.

Fagonia indica Burm.f. is known for its anti-infective character and has been studied in the present work as a synergistic remedy against resistant bacterial strains. Initially, phytochemicals were quantified in n-Hexane (n-Hex), ethyl acetate (E.A), methanol (MeOH), and aqueous (Aq.) extracts by Total Phenolic Content (TPC), Total Flavonoid Content (TFC) and Reverse Phase High Performance Liquid Chromatography (RP-HPLC) analysis. Later, after establishing an antibacterial resistance profile for extracts and antibiotics against gram-positive and gram-negative strains, synergism was evaluated in combination with cefixime through time-kill kinetics and bacterial protein estimation studies. Topographic images depicting synergism were obtained by scanning electron microscopy for Methicilin-resistant Staphylococcus aureus (MRSA) and Resistant Escherichia coli (R.E. coli). Results showed the presence of maximum phenolic (28.4 ± 0.67 µg GAE/mg extract) and flavonoid (11 ± 0.42 µg QE/mg extract) contents in MeOH extract. RP-HPLC results also displayed maximum polyphenols in MeOH extract followed by E.A extract. Clinical strains were resistant to cefixime whereas these were moderately inhibited by all extracts (MIC 150-300 µg/ml) except Aq. extract. E.A and n-Hex extracts demonstrated maximum synergism (Fractional inhibitory concentration index (FICI) 0.31) against R.E. coli. The n-Hex extract displayed total synergism against R.P. a with a 4-fold reduction in cefixime dose. Time-kill kinetics showed maximum inhibition of gram-negative bacterial growth from 3 to 12 h when treated at FICI and 2FICI values with > 10-fold reduction of the extracts' dose. All combinations demonstrate > 70 % protein content inhibition with bacterial cell wall disruption in SEM images. Fortunately, FICI concentrations have low hemolytic potential (<5%). Conclusively, F. indica extracts can mitigate antimicrobial resistance against cefixime and can be investigated in detail by in vivo and mechanistic studies.

Epithelioid hepatic angiomyolipoma in pregnancy: A case report.

INTRODUCTION AND IMPORTANCE: Epithelioid hepatic angiomyolipoma (HAML) is a rare benign tumor predominantly found in women. Its occurrence during pregnancy is extremely rare. Accurate diagnosis of HAML is challenging due to its radiological resemblance to other hepatic neoplasms. We present a case of epithelioid HAML in a pregnant patient, highlighting the diagnostic and management challenges encountered. CASE PRESENTATION: A 24-year-old pregnant female, in her fifth month of pregnancy, presented with right hypochondrium pain and nausea. Radiological imaging suggested the possibility of a hepatic adenoma. The patient opted to continue the pregnancy with regular monitoring of the mass as well as fetal health. After delivering a healthy baby, the patient underwent successful mass excision and cholecystectomy. Histopathology of the liver mass confirmed the diagnosis of epithelioid HAML. CLINICAL DISCUSSION: Epithelioid HAML is a rare tumor often misdiagnosed. It is more aggressive and frequently associated with tuberous sclerosis complex (TSC) compared to other subtypes. The diagnosis of HAML can be challenging due to its resemblance to Hepatocellular Carcinoma and other hepatic neoplasms on radiological imaging. Immunohistochemistry plays a crucial role in confirming the diagnosis. Surgical excision is the recommended treatment, with complete removal to minimize the risk of recurrence. CONCLUSION: This case report highlights the rarity of epithelioid HAML during pregnancy and emphasizes the importance of a multidisciplinary approach in managing hepatic neoplasms. Close monitoring is crucial, considering the potential risks to the mother and fetus. Accurate diagnosis through histopathological evaluation, immunohistochemistry and a multidisciplinary approach are essential for appropriate management.

Impact of sublethal concentrations of flonicamid on key demographic parameters and feeding behavior of Schizaphis graminum.

Flonicamid is a novel systemic insecticide that efficiently controls sap-sucking insect pests. However, the impact of sublethal concentrations of flonicamid on key demographic parameters and the feeding behavior of greenbug, Schizaphis graminum has not yet been studied. In this study, we used the age stage, two-sex life table approach, and electrical penetration graphs (EPGs) to investigate the sublethal effects of flonicamid on the biological traits and feeding behavior of S. graminum. Bioassays showed that flonicamid possesses high toxicity to adult S. graminum with LC50 of 5.111 mg L-1 following 48 h exposure. Sublethal concentrations of flonicamid (LC5 and LC10) significantly decreased the longevity and fecundity of directly exposed parental aphids (F0), while the reproductive days were reduced only at LC10. The pre-adult stage and total pre-reproductive period (TPRP) increased in F1 individuals after exposure of F0 aphids to the sublethal concentrations of flonicamid. Furthermore, the adult longevity, fecundity and key demographic parameters (R0, r, and λ) were significantly reduced in progeny generation (F1). EPG recordings showed that the total duration of phloem sap ingestion and concurrent salivation (E2) decreased substantially in F0 and F1 aphids after exposure to LC5 and LC10 of flonicamid. Taken together, our results showed that the sublethal concentrations of flonicamid affect the demographic parameters and feeding behavior that ultimately suppress the population growth of S. graminum. This study provides in-depth information about the overall effects of flonicamid on S. graminum that might help to manage this key pest.

Neuroprotective mechanism of Ajugarin-I against Vincristine-Induced neuropathic pain via regulation of Nrf2/NF-κB and Bcl2 signalling.

Vincristine (VCR) is a well-known chemotherapeutic agent that frequently triggers neuropathic pain. Ajugarin-I (Aju-I) isolated from Ajuga bracteosa exerts antioxidant, anti-inflammatory, and neuroprotective properties. The present study was designed to investigate the ameliorative potential of Aju-I against VCR-induced neuropathic pain and explored the underlying mechanism involved. The neuroprotective potential of Aju-I was first confirmed against hydrogen peroxide (H2O2)-induced cytotoxicity and oxidative stress in PC12 cells. For neuropathic pain induction, vincristine was given intraperitoneally (i.p.) into adult male albino mice (BALB/c) of the same age (8-12 weeks old) for 10 days (days 1-10). Aju-I (1 and 5 mg/kg) doses were administered from day 11 to 21 intraperitoneally (i.p.) after the neuropathic induction. Initially, behavioral tests such as thermal hyperalgesia, mechanical allodynia, and cold allodynia were performed to investigate the antinociceptive potential of Ajugarin-I (1 and 5 mg/kg, b.w). The nuclear factor-erythroid factor 2-related factor 2(Nrf2), nuclear factor-κB (NF-κB), BCL2-associated × protein (Bax), and B-cell-lymphoma-2 (Bcl-2) signaling proteins were determined by immunohistochemistry and western blot. Additionally, inflammatory cytokines, antioxidant, and oxidative stress parameters were also measured in the spinal cord and sciatic nerve. The behavioral results demonstrated that Aju-I (5 mg/kg) markedly alleviated VCR-induced neuropathic pain behaviors including hyperalgesia and allodynia. It reversed the histological alterations caused by VCR in the sciatic nerve, spinal cord, and brain. It significantly alleviated oxidative stress and inflammation by regulating the immunoreactivity of Nrf2/NF-κB signaling. It suppressed apoptosis by regulating the immunoreactivity of Bcl-2/Bax and Caspase-3. The flow cytometry and comet analysis also confirmed its anti-apoptotic potential. It considerably improved the antioxidant status and mitigated VCR-induced inflammatory cytokines. High-performance liquid chromatography (HPLC) analysis indicated that Aju-I crosses the blood-brain barrier (BBB) and penetrated the brain tissue. These findings suggest that Aju-I treatment inhibited vincristine-induced neuropathy via regulation of Nrf2/NF-κB and Bcl2 signaling.

Coagulansin-A improves spatial memory in 5xFAD Tg mice by targeting Nrf-2/NF-κB and Bcl-2 pathway.

The present study was designed to investigate the underlying molecular signaling of Coagulansin-A (Coag-A) as a therapeutic agent against Alzheimer's disease (AD). Preliminarily, it exhibited a neuroprotective effect against H2O2-induced oxidative stress in HT-22 cells. The in vivo studies were performed by administering Coag-A (0.1, 1, and 10 mg/kg) intraperitoneally to 5xFAD transgenic (Tg) mouse model. Coag-A (10 mg/kg) significantly attenuated the cognitive decline compared to Tg mice group in the shallow water maze (SWM) and Y-maze test paradigms. The anti-aggregation potential of Coag-A was determined by performing Fourier transform-infrared (FT-IR) spectroscopy and differential scanning calorimeter (DSC) analysis in the prefrontal cortex (PFC) and hippocampal (HC) regions of mice brain. The FT-IR spectra demonstrated the inhibition of amyloid beta (Aß) through a decrease in ß-sheet aggregation, along with the inhibition of changes in the lipids, proteins, and phospholipids. The DSC analysis displayed a low-temperature exotherm associated with the reversible process of aggregation of soluble protein fractions prior to denaturation. Furthermore, Coag-A treatment displayed a regular density of granule cells in H&E stained sections, along with a reduced amyloid load and PAS-positive granules in all the regions of interest in mice brain. The real-time polymerase chain reaction (q-PCR), western blot and immunohistochemical (IHC) analysis demonstrated antioxidant, anti-inflammatory, and anti-apoptotic effect of Coag-A by enhancing the expression of nuclear factor erythroid-2-related factor (Nrf-2) and reducing nuclear factor kappa B (NF-κB) and Bax protein expression. In addition, Coag-A significantly increased the antioxidant enzymes and proteins level, along with a reduced pro-inflammatory cytokines production.

Suppression of TRPV1/TRPM8/P2Y Nociceptors by Withametelin via Downregulating MAPK Signaling in Mouse Model of Vincristine-Induced Neuropathic Pain.

Vincristine (VCR) is a widely used chemotherapy drug that induced peripheral painful neuropathy. Yet, it still lacks an ideal therapeutic strategy. The transient receptor potential (TRP) channels, purinergic receptor (P2Y), and mitogen-activated protein kinase (MAPK) signaling play a crucial role in the pathogenesis of neuropathic pain. Withametelin (WMT), a potential Phytosteroid isolated from datura innoxa, exhibits remarkable neuroprotective properties. The present investigation was designed to explore the effect of withametelin on VCR-induced neuropathic pain and its underlying molecular mechanism. Initially, the neuroprotective potential of WMT was confirmed against hydrogen peroxide (H2O2)-induced PC12 cells. To develop potential candidates for neuropathic pain treatment, a VCR-induced neuropathic pain model was established. Vincristine (75 µg/kg) was administered intraperitoneally (i.p.) for 10 consecutive days (day 1-10) for the induction of neuropathic pain. Gabapentin (GBP) (60 mg/kg, i.p.) and withametelin (0.1 and 1 mg/kg i.p.) treatments were given after the completion of VCR injection on the 11th day up to 21 days. The results revealed that WMT significantly reduced VCR-induced pain hypersensitivity, including mechanical allodynia, cold allodynia, and thermal hyperalgesia. It reversed the VCR-induced histopathological changes in the brain, spinal cord, and sciatic nerve. It inhibited VCR-induced changes in the biochemical composition of the myelin sheath of the sciatic nerve. It markedly downregulated the expression levels of TRPV1 (transient receptor potential vanilloid 1); TRPM8 (Transient receptor potential melastatin 8); and P2Y nociceptors and MAPKs signaling, including ERK (Extracellular Signal-Regulated Kinase), JNK (c-Jun N-terminal kinase), and p-38 in the spinal cord. It suppressed apoptosis by regulating Bax (Bcl2-associated X-protein), Bcl-2 (B-cell-lymphoma-2), and Caspase-3 expression. It considerably attenuated inflammatory cytokines, oxidative stress, and genotoxicity. This study suggests that WMT treatment suppressed vincristine-induced neuropathic pain by targeting the TRPV1/TRPM8/P2Y nociceptors and MAPK signaling.

Synthesis, antioxidant, antimicrobial and antiviral docking studies of ethyl 2-(2-(arylidene)hydrazinyl)thiazole-4-carboxylates.

A series of ethyl 2-(2-(arylidene)hydrazinyl)thiazole-4-carboxylates (2a-r) was synthesized in two steps from thiosemicarbazones (1a-r), which were cyclized with ethyl bromopyruvate to ethyl 2-(2-(arylidene)hydrazinyl)thiazole-4-carboxylates (2a-r). The structures of compounds (2a-r) were established by FT-IR, 1H- and 13C-NMR. The structure of compound 2a was confirmed by HRMS. The compounds (2a-r) were then evaluated for their antimicrobial and antioxidant assays. The antioxidant studies revealed, ethyl 2-(2-(4-hydroxy-3-methoxybenzylidene)hydrazinyl)thiazole-4-carboxylate (2g) and ethyl 2-(2-(1-phenylethylidene)hydrazinyl)thiazole-4-carboxylate (2h) as promising antioxidant agents with %FRSA: 84.46 ± 0.13 and 74.50 ± 0.37, TAC: 269.08 ± 0.92 and 269.11 ± 0.61 and TRP: 272.34 ± 0.87 and 231.11 ± 0.67 µg AAE/mg dry weight of compound. Beside bioactivities, density functional theory (DFT) methods were used to study the electronic structure and properties of synthesized compounds (2a-m). The potential of synthesized compounds for possible antiviral targets is also predicted through molecular docking methods. The compounds 2e and 2h showed good binding affinities and inhibition constants to be considered as therapeutic target for Mpro protein of SARS-CoV-2 (COVID-19). The present in-depth analysis of synthesized compounds will put them under the spot light for practical applications as antioxidants and the modification in structural motif may open the way for COVID-19 drug.

Development and pharmacological evaluation of vancomycin loaded chitosan films.

Burn injuries are the most prevalent and devastating form of skin trauma. Current study aimed to fabricate novel chitosan-based composite films of vancomycin for wound healing applications. The developed vancomycin-chitosan films were evaluated for various quality attributes and were subjected to anti-bacterial activity against methicillin resistant Staphylococcus aureus (MRSA) and wound healing efficacy study in rat model. The prepared vancomycin-chitosan film 2 (VCF2) physically displayed a substantial tensile strength and swelling ratio. Pharmacologically, VCF2 exhibited sustained vancomycin release, excellent antibacterial activity and improved wound healing efficacy in rats. The superior wound healing potential was ascribed to the enhanced levels of reduced glutathione, glutathione-S-transferase, catalase and decreased lipid peroxidation. Furthermore, improved angiogenesis, granulation, epidermal regeneration and down regulation in the expressions of tumor necrosis factor, cyclooxygenase-2 and nuclear factor kappa B were the reasons of improved wound healing as confirmed by histopathological and molecular techniques. Thus, it is plausible to say that VCF2 could provide a potential therapeutic approach in burn wounds.

Red light controlled callus morphogenetic patterns and secondary metabolites production in Withania somnifera L.

Withania somnifera L. is an endangered medicinal plant of higher market value. The in vitro callus cultures were established on Murashige and Skoog (MS) media augmented with different plant growth regulators. The MS medium containing 0.5 mg∙L-1 of each TDZ and NAA was found to be optimal for callus formation and growth. Further, callus cultures were raised in different light wavelengths to find the right wavelength carrying the photons for the ideal cell growth of W. somnifera. Among the different wavelengths, red light was best for maximum biomass accumulation in callus culture. However, violet light condition was proven to be favouring the phenols and flavonoids synthesis in the callus cultures. Compared to other wavelengths, red light grown callus extract showed significantly higher content of chlorogenic acid, and withaferin A. This study concludes that red light treatment was optimum for maximum biomass accumulation and anti-oxidant activity in calli of W. somnifera.

Numerical Investigation of 1 × 7 Steel Wire Strand under Fretting Fatigue Condition.

Wire ropes undergo a fretting fatigue condition when subjected to axial and bending loads. The fretting behavior of wires are classified as line contact and trellis point of contact. The experimental study on the fatigue of wire ropes indicates that most of the failure occurs due to high localized stresses at trellis point of contact. A continuum damage mechanics approach was previously proposed to estimate the fatigue life estimation of wire ropes. The approach majorly depends on the high value of localized stresses as well as the micro-slippage occurs at the contact region. Finite element approach has been used to study radial and axial distribution of stresses and displacement in order to clearly understand the evolution of stresses and existence of relative displacements between neighboring wires under various loading and frictional conditions. The relative movements of contacting wires are more when friction is not considered. In the presence of friction, the relative movement occurs at the boundaries of the contact region. The location of microslip in the presence of friction is backed by the experimental observation stating the crack is initiated at or the outer boundary of the contact spot. The existence of slip is due to different displacement of outer and central wires.

Comprehensive investigations on anti-leishmanial potentials of Euphorbia wallichii root extract and its effects on membrane permeability and apoptosis.

Clinically available synthetic chemotherapeutics to treat the vector-borne protozoan infection, leishmaniasis, are associated with serious complications such as toxicity and emergence of resistance. Natural products from plants consist of interesting biomolecules that may interfere with DNA or membrane integrity of the parasite and can possibly minimise the associated side effects. In the present study, various fractions of Euphorbia wallichii (EW) root extracts including n-hexane (EWNX), ethyl acetate (EWEA), chloroform (EWCH) and aqueous (EWAQ), were evaluated for their antileishmanial potential against Leishmania tropica followed by investigation of the possible mechanism of action via reactive oxygen species (ROS) quantification, membrane permeability (via sytox green dye) and apoptotic assay (via AO/EB method) using fluorescent microscopy. Two of the fractions i.e. EWEA and EWAQ inhibited the growth of promastigotes (IC50 7.8 and 10.2 µg/mL, respectively) and amastigotes (IC50 9.9 and 13.3 µg/mL, respectively) forms almost at similar concentrations as found for the standard antileishmanial drugs, tartar emetic (TA) and Glucantime (IC50 9.4 and 21.5 µg/mL, respectively). Both the active fractions remained non-toxic towards human blood erythrocytes and were able to cause membrane permeability and apoptotic induction (using Triton X-100 as a positive control) leading to death of Leishmania parasites. However, both the fractions could not triger significant and persistent ROS generation, compared to hydrogen peroxide used as a positive control. Antilesihmanial activity of the two active fractions might be attributed to the presence of high quantity of tannins and saponins.

Comparison of classical and transgenic genetic sexing strains of Mediterranean fruit fly (Diptera: Tephritidae) for application of the sterile insect technique.

The development of genetic sexing strains (GSSs) based on classical genetic approaches has revolutionized the application of the sterile insect technique (SIT) against the Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). The global use of Mediterranean fruit fly GSS for SIT applications as part of area-wide integrated pest management (AW-IPM) programmes is testimony to their effectiveness. During recent years, transgenic sexing strains (TSSs) have been developed through genetic engineering techniques offering the possibility to produce male-only progeny by introducing female embryonic lethal genes and to increase the efficacy to identify released sterile males by means of the expression of fluorescent transgene markers. Here, we present a comparative analysis of two Mediterranean fruit fly strains: the classical GSS VIENNA 8D53-/Toliman and the transgenic FSEL#32. The strains were compared for production efficiency and quality control indices under semi mass-rearing conditions, response to sterilizing irradiation doses, male mating performance in walk-in field cages, and production cost of male-only pupae. The results showed that, the FSEL #32 TSS had a similar fecundity but a higher production of male-only pupae than the VIENNA 8D53-/Toliman GSS. For some of the quality control parameters tested, such as pupal weight and survival under starvation conditions, the FSEL #32 TSS was inferior to the VIENNA 8D53-/Toliman GSS. Both the transgenic and the classical genetic sexing strains have shown acceptable and similar mating competitiveness when compared with wild males for mating with wild females. The cost production for both strains is similar but the FSEL#32 TSS may potentially be more cost effective at higher production levels. The results are discussed in the context of incorporating the transgenic strain for SIT application.

Correction to: Implication of OPRM1 A118G Polymorphism in Opioids Addicts in Pakistan: In vitro and In silico Analysis.

The original version of this article is missing the Acknowledgments section.

Implication of OPRM1 A118G Polymorphism in Opioids Addicts in Pakistan: In vitro and In silico Analysis.

Single nucleotide polymorphism in OPRM1 gene is associated with hedonic and reinforcing consequences of opioids. Risk and protective alleles may vary in different populations. One hundred healthy controls and 100 opioids (predominantly heroin) addicts from Pakistani origin were genotyped for A118G (N40D) polymorphism in OPRM1. Structural and functional impact of the polymorphism on encoded protein was predicted by in silico analysis. Results show significant association between homozygous GG genotype and opioid addiction in Pakistani population (p value = 0.016). In silico analysis by SIFT (TI = 0.61), PolyPhen (PISC = 0.227), PANTHER (subPSEC = -1.7171), and SNP effect predicted this SNP benign for encoded protein. Superimposing wild-type and mutated proteins by MODELLER shows no change (RMSD = 0.1) in extracellular ligand binding domain of µ-opioid receptor. However, Haploreg and RegulomeDB predicted OPRM1 gene repression by chromatin condensation and increased binding affinity of RXRA transcription factor that may reduce protein translation and hence the number of available receptors to bind with drugs, which may trigger underlying mechanisms for opioids addiction. Thus, this study outlines causal relationship between opioids addiction and genetic predisposition in Pakistani population.

Cytotoxicity, In vitro anti-Leishmanial and fingerprint HPLC- photodiode array analysis of the roots of Trillium govanianum.

Trillium govanianum Wall. ex D. Don (Melanthiaceae alt. Trilliaceae), commonly known as 'nagchhatry' or 'teen patra', distributed from Pakistan to Bhutan about 2500-3800 m altitude is indigenous to Himalayas region. In folk medicine the plant has been reported for the treatment of wound healing, sepsis and in various sexual disorders. This paper reports, for the first time, to evaluate the cytotoxicity, in vitro anti-leishmanial (promastigotes) and fingerprint HPLC-photodiode array analysis of the MeOH extract of the roots of T. govanianum and its solid phase extraction fractions. Reverse phase HPLC-PDA based quantification revealed the presence of significant amount of quercetin, myrecetin and kaemferol ranging from 0.221to 0.528 µg/mg DW. MeOH extract revealed distinguishable protein kinase inhibitory activity against Streptomyces 85E strain with 18 mm bald phenotype. The remarkable toxicity profile against brine shrimps and leishmanial was manifested by MeOH extract with LC50 10 and 38.5 µg/mL, respectively.

Automated System for Referral of Cotton-Wool Spots.

BACKGROUND: Cotton-wool spots also referred as soft exudates are the early signs of complications in the eye fundus of the patients suffering from diabetic retinopathy. Early detection of exudates helps in the diagnosis of the disease and provides better medical attention. METHODS: In this paper, an automated system for the detection of soft exudates has been suggested. The system has been developed by the combination of different techniques like Scale Invariant Feature Transform (SIFT), Visual Dictionaries, K-means clustering and Support Vector Machine (SVM). RESULTS: The performance of the system is evaluated on a publically available dataset and AUC of 94.59% is achieved with the highest accuracy obtained is 94.59%. The experiments are also performed after mixing three datasets and AUC of 92.61% is observed with 91.94% accuracy. CONCLUSION: The proposed system is easy to implement and can be used by medical experts both online and offline for referral of Cotton-wool spots in large populations. The system shows promising performance.

Kinnow peel extract as a reducing and capping agent for the fabrication of silver NPs and their biological applications.

An effective approach used for the synthesis of silver nanoparticles (AgNPs) through green chemistry by using Kinnow peel extract as a reducing and capping agent is presented. Two different approaches, diluted and concentrated peel extracts, were used for the synthesis of AgNPs. Ultraviolet-visible spectroscopy exhibits characteristic absorption peaks at 425 and 400 nm for nanoparticles (NPs) synthesised by diluted and concentrated extracts, respectively. The X-ray diffraction analysis of nanofabricated silver exhibited a pure face centred cubic structure of 27.4 and 18.1 nm sizes calculated by using Scherrer equation. Scanning electron microscopy analysis showed a uniform morphology of synthesised NPs. Significant antioxidant, phytochemical and antibacterial assays show that both AgNPs can be effectively used in biomedical applications. Furthermore, the use of citrus peel for the synthesis of NPs can be an effective tool in waste management.

ZnO nanostructure fabrication in different solvents transforms physio-chemical, biological and photodegradable properties.

Zinc oxide (ZnO) nanostructures are synthesized in various organic solvents (acetone, chloroform, ethyl acetate, ethanol and methanol) and water via coprecipitation process using zinc acetate as precursor. The resultant ZnO nanoparticles, nano rods and nano sheets are characterized by UV-vis spectrophotometric analysis, scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transmission infrared spectroscopy (FTIR), and energy dispersive X-ray spectroscopy (EDX). The variable size and geometry of nanoparticles depend upon medium used for synthesis. The synthesized ZnO nanostructures exhibit minor to moderate antioxidative (DPPH based free radical scavenging activity, total antioxidative potential and total reducing power) response. Mild to moderate antibacterial and antifungal activities, excellent antileishmanial potential (IC50 up to 3.76), and good cytotoxic perspective (LD50 up to 49.4) is also observed by the synthesized ZnO NPs. The nanoparticles also exhibit moderate α-amylase inhibition response. Furthermore the nanostructures are evaluated for methylene blue photodegradation response within 60min time period. It is found that organic solvent alters shape, size and other physio-chemical properties of ZnO that ultimately modulate the biological, chemical, and environmental properties.