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Nargis Aman, Khalid Rauf, Shujaat Ali Khan, Ahmed Tokhi, Naeem-Ur Rehman, Muhammad Arfat YameenDepartment of Pharmacy COMSATS University Islamabad, Abbottabad Campus, Abbottabad, KPK, PakistanPurpose: To investigate the effects of zinc oxide nanoparticles (ZnO NPs) on chloroquine (CQ)-induced itching, and overall behavior of mice after oral administration of ZnO NPs of various sizes and doses.Background: With the wide-spread use of ZnO NPs in pharmaceuticals and cosmetics, concerns about their safety and toxicity are also increasing. Multiple aspects of ZnO NPs regarding cytotoxicity and tolerability are under investigation globally. Still, a clear conclusion about their safety has not been reached. Chloroquine phosphate is an antimalarial with known side effects of itching in humans and animals. In this study, CQ was used to induce itching in mice, and the effects of ZnO NPs on scratching and other neurological behavior of mice were observed.Methods: Female BALB/c mice were divided into eleven groups of six mice each. ZnO NPs of various sizes and doses were administered orally 1 hour before CQ (32 mg/kg body weight) was administered subcutaneously. The effect of ZnO NPs on CQ-induced pruritus was observed for the next 30 minutes. Simultaneously, overall behavioral changes (socialization and locomotion) were also recorded using a video camera.Results: A significant reduction (P˂0.001) in scratching bouts was observed at all three doses of ZnO NPs (particle sizes 100, 30 nm, and green synthesized 30 nm). Locomotion was reduced significantly (P˂0.001) in ZnO NPs-treated groups in comparison to normal saline and CQ group, additionally, a significant increase in socialization (P˂0.05) was observed in ZnO NP-treated groups as compared to CQ group.Conclusion: ZnO NPs, instead of aggravating the dermatological condition, ameliorated the pruritus. All sizes of ZnO NPs used significantly improved socialization among mice and reduced locomotion activity.Keywords: ZnO NPs, chloroquine-induced pruritus, CQ, green synthesized ZnO NPs, mice

Background The Global Polio Eradication Initiative (GPEI) helped develop the standard acute flaccid paralysis surveillance (AFP) system worldwide, including, knowledge, expertise, technical assistance, and trained personnel. AFP surveillance can complement any disease surveillance system. Objective This study outlines AFP surveillance evolution in Bangladesh, its success and challenging factors, and its potential to facilitate other health goals. Methods This mixed-method study includes a grey literature review, survey, and key informant interviews (KIIs). We collected grey literature from online websites and paper documentation from GPEI stakeholders. Online and in-person surveys were conducted in six divisions of Bangladesh, including Dhaka, Rajshahi, Rangpur, Chittagong, Sylhet, and Khulna, to map tacit knowledge ideas, approaches, and experiences. We also conducted KIIs, and Data were then combined on focused emerging themes, including the history, challenges, and successes of AFP surveillance programme. Results According to the grey literature review, survey, and KII, AFP surveillance successfully contributed to decreasing polio in Bangladesh. The major facilitating factors were multi-sectoral collaboration, Surveillance Immunization Medical Officer (SIMO) network activities, social environment, community-based surveillance, and promising political commitment. On the other hand, high population growth, hard-to-reach areas, people residing in risky zones, and polio transition planning were significant challenges. Bangladesh is also utilizing these polio surveillance assets for other vaccine-preventable diseases. Conclusion As the world is so close to eradicating polio, the knowledge, and other assets of the AFP surveillance, could be used for other health programmes. In addition, its strengths can be leveraged for combating new and emerging diseases.

Abstract Background Kangaroo mother care (KMC) is a proven low-cost intervention to prevent neonatal mortality of pre-term and low birth weight babies and is very relevant to Bangladesh. KMC provides thermal regulation and thus directly avert neonatal mortality. KMC includes early, continuous, and prolonged skin-to-skin contact between an infant and caregiver, exclusive breastfeeding, early discharge from the hospital, and post-discharge follow-up. The purpose of this study was to investigate the fidelity of this intervention’s implementation according to national guidelines across all tiers of government (public) health facilities of Bangladesh. Methods We adopted a triangulation mixed-methods approach of both quantitative and qualitative components in this research to support and explain the information obtained from quantitative observation with the help of qualitative interviews on the fidelity of KMC practice. We used an observation checklist to find the fidelity of KMC practice and used semi-structured guidelines to explain and understand the moderators of fidelity through key informant interviews and in-depth interviews. We undertook eight facility visits in four districts, observed twenty-three neonates and their caregivers during KMC practice at those facilities, and conducted twenty-seven key informant interviews with facility managers, health care providers, and five in-depth interviews with caregivers. Extracted information was triangulated and arranged under the themes of the fidelity framework. Results Despite being a low-cost intervention, findings exhibit some adherence to the national guideline with several gaps in practice. Leadership played a critical role in ensuring the KMC practice. Specific components of KMC practice, like duration, nutrition maintenance, discharge criteria, and follow-up, were not consistent as recommended. Infrastructure, human resources, developmental partner support, and the demand-side and supply-side responsiveness played a critical role in enacting this human-centric approach’s fidelity. The observed interruption found in the implementation process posed threats to achieve the intended outcome as these caused violations of the basic principles of KMC. Conclusions The study findings will help find ways to effectively deliver this intervention so that fidelity of practice is maintained, enhancing KMC services’ quality and advocating towards the successful scale-up of this program.

Vesicovaginal reflux is a common cause of urinary incontinence in girls. A micturating cystourethrogram, which is the diagnostic investigation of choice, can demonstrate retrograde filling of the vagina during micturition and the complete emptying of the vagina at the end of micturition. Vesicovaginal reflux is a rare cause of gross hydrocolpos occurring without any anatomical obstruction. The condition may be associated with functional voiding disturbances.

Abstract Background Intracranial stenosis is the most common cause of stroke among Asians. It has a poor prognosis with a high rate of recurrence. No effective medical or surgical treatment modality has been developed for the treatment of stroke due to intracranial stenosis. We aim to identify risk factors and biomarkers for intracranial stenosis and to develop techniques such as use of transcranial doppler to help diagnose intracranial stenosis in a cost-effective manner. Methods/Design The Karachi Intracranial Stenosis Study (KISS) is a prospective, observational, case-control study to describe the clinical features and determine the risk factors of patients with stroke due to intracranial stenosis and compare them to those with stroke due to other etiologies as well as to unaffected individuals. We plan to recruit 200 patients with stroke due to intracranial stenosis and two control groups each of 150 matched individuals. The first set of controls will include patients with ischemic stroke that is due to other atherosclerotic mechanisms specifically lacunar and cardioembolic strokes. The second group will consist of stroke free individuals. Standardized interviews will be conducted to determine demographic, medical, social, and behavioral variables along with baseline medications. Mandatory procedures for inclusion in the study are clinical confirmation of stroke by a healthcare professional within 72 hours of onset, 12 lead electrocardiogram, and neuroimaging. In addition, lipid profile, serum glucose, creatinine and HbA1C will be measured in all participants. Ancillary tests will include carotid ultrasound, transcranial doppler and magnetic resonance or computed tomography angiogram to rule out concurrent carotid disease. Echocardiogram and other additional investigations will be performed at these centers at the discretion of the regional physicians. Discussion The results of this study will help inform locally relevant clinical guidelines and effective public health and individual interventions.

Biofilms are complex communities of microorganisms that are enclosed in a matrix that shows increased resistance to antimicrobial and immunological encounters. Mostly, the traditional methods to control biofilm are exhausted; therefore, the aim is to evaluate the potential of essential oil (EO) from Tagetes minuta to encounter biofilm and other related virulence factors. The EO of T. minuta was extracted through steam-distillation, analyzed on gas chromatography-mass spectrometry, and the biofilm inhibition assays were performed with various concentrations of EO. Mainly the EO from T. minuta contains cis -β-ocimene (29.1%), trans -tagetenone (23.1%), and cis -tagetenone (17.7%). The virulence factors were monitored while applying different concentrations of EO and it was recorded that the EO from T. minuta significantly inhibited the virulence factors linked with quorum sensing (QS), such as pyocyanin production, protease production, and swarming motility. Biofilm formation is one of the most important virulence factors associated with the QS pathway and was inhibited up to 79% in the presence of EO. Antibacterial activity against the PAO1 of EO was not so promising particularly and it has high MIC (325 μg/mL) and MBC (5000 μg/mL). EO is quite efficient to inhibit biofilm in a very small concentration of 20 μg/mL, which confirms that the biofilm inhibition by EO is not by killing bacterial cells but by inhibiting the QS pathway. The study on PAO1 constructs carrying various QS reported genes confirmed that the EO interferes with the QS pathway that ultimately controls various virulence factors caused by PAO1., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Joint defects associated with a variety of etiologies often extend deep into the subchondral bone leading to functional impairment and joint immobility, and it is a very challenging task to regenerate the bone-cartilage interface offering significant opportunities for biomaterial-based interventions to improve the quality of life of patients. Herein drug-/bioactive-loaded porous tissue scaffolds incorporating nano-hydroxyapatite (nHAp), chitosan (CS) and either hydroxypropyl methylcellulose (HPMC) or Bombyx mori silk fibroin (SF) are fabricated through freeze drying method as subchondral bone substitute. A combination of spectroscopy and microscopy (Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), energy dispersive X-ray (EDX), and X-ray fluorescence (XRF) were used to analyze the structure of the porous biomaterials. The compressive mechanical properties of these scaffolds are biomimetic of cancellous bone tissues and capable of releasing drugs/bioactives (exemplified with triamcinolone acetonide, TA, or transforming growth factor-β1, TGF-β1, respectively) over a period of days. Mouse preosteoblast MC3T3-E1 cells were observed to adhere and proliferate on the tissue scaffolds as confirmed by the cell attachment, live-dead assay and alamarBlue™ assay. Interestingly, RT-qPCR analysis showed that the TA downregulated inflammatory biomarkers and upregulated the bone-specific biomarkers, suggesting such tissue scaffolds have long-term potential for clinical application., Competing Interests: Declaration of competing interest The authors declare no conflict of interest. Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

This study aimed to synthesize the silver nanoparticles (SNPs) and loaded chitosan nanoparticles (LCNPs) using Euphorbia prostata based on their anticandidal activity. Antioxidant capacity and the total phenolic and total flavonoid content of plant samples and synthesized nanoparticles (NPs) were also evaluated. SNPs and LCNPs were prepared, respectively using chemical reduction of silver salt solution and ionotropic gelation method. The anticandidal activity was assessed by broth micro-dilution method and the antioxidant activity was determined using free-radical scavenging assays. The synthesized NPs after the optimization process were found to be spherical with sizes ranging from 12 to 100 nm. Spectroscopic analysis of NPs showed the appearance of peaks in prescribed wavelength ranging between 402 and 493 nm. The synthesized NPs showed potent anticandidal activity compared to the free extract. The SNPs formulations NpEPM 7.5 and NpEPMR 7.5, showed significantly low MIC values ranging between 2 and 128 µg/mL. In the case of LCNPs, NpEPM (4:1) and NpEPME (4:1) also showed lower MIC values ranging from 32 to 256 µg/mL. The plant samples as well as NPs showed antioxidant potential. In addition, plant extracts and NPs possess the potent biological potential and can be further investigated through in vivo experiments.

Implantation of biomaterials capable of the controlled release of antibacterials during articular cartilage repair may prevent postoperative infections. Herein, biomaterials are prepared with biomimetic architectures (nonwoven mats of fibers) via electrospinning that are composed of poly(ɛ-caprolactone), poly(lactic acid), and Bombyx mori silk fibroin (with varying ratios) and, optionally, an antibiotic drug (cefixime trihydrate). The composition, morphology, and mechanical properties of the nanofibrous mats are characterized using scanning electron microscope, Fourier transform infrared spectroscopy, and tensile testing. The nonwoven mats have nanoscale fibers (typical diameters of 324-725 nm) and are capable of controlling the release profiles of the drug, with antibacterial activity against Gram +ve and Gram -ve bacteria (two common strains of human pathogenic bacteria, Staphylococcus aureus and Escherichia coli) under in vitro static conditions. The drug loaded nanofiber mats display cytocompatibility comparable to pure poly(ɛ-caprolactone) nanofibers when cultured with National Institutes of Health (NIH) NIH-3T3 fibroblast cell line and have long-term potential for clinical applications in the field of pharmaceutical sciences., (© 2022 The Authors. Macromolecular Bioscience published by Wiley-VCH GmbH.)

Phragmanthera capitata is a medicinal plant used in traditional medicine to treat several diseases, including diabetes. Its antioxidant properties and inhibitory effects on enzyme-carbohydrate digestion activities have been demonstrated. The present study aimed to provide data that can contribute to rationalizing the preparation of antioxidant and antidiabetic extracts from this plant. P. capitata (whole plant) growing on Persea americana was harvested at the fruiting stage. A response surface design-type central composite was applied to maximize the extraction yield, phenolic contents, and antioxidant and antidiabetic properties of the ethyl acetate extract of P. capitata . The influencing extraction factors were temperature, powder particle size, and solvent-to-powder ratio. The total phenolic content, total antioxidant capacity (TAC), DPPH scavenging ability, ferric reducing antioxidant power (FRAP), and antidiabetic ( α -amylase and α -glucosidase inhibitory) effects of the extracts were determined using conventional methods. A temperature above 55°C contributed to the degradation of the extract, which was reflected in the GC-MS profile by a significant reduction in the number of compounds it contained. The optimal conditions were defined as 24.42°C for temperature, 250  µ m powder particle size, and 8.30 (v:w) solvent-to-powder ratio. This extraction protocol resulted in more than twice the extraction yield (3.05%), TTC (62.30 mg TAE/g), TAC (41.41 mg AAE/g), FRAP (186.56 mg AAE/g), and α -amylase (IC 50 15.05  µ g/mL) and α -glucosidase (IC 50 21.14  µ g/mL) inhibitory activities compared to our previous results. Additionally, these optimal conditions led experimentally to the extraction of higher phenolic content and to the attainment of higher antioxidant and antidiabetic activity, which closely matched the predicted values. Using these conditions, it is possible to prepare an antidiabetic phytomedicine from P. capitatathat can prevent oxidative stress complications. However, further complementary studies should be carried out considering other factors that influence the composition and pharmacological properties of the extract., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2022 Césaire Feudjio et al.)

Increasing resistance to existing antibiotics by microbes is currently the biggest dilemma. Antimicrobial photodynamic therapy is a promising alternative for the treatment of multidrug-resistant infections. The aim of the current study was to fabricate graphene quantum dots loaded with curcumin as photosensitizer for improved antimicrobial photodynamic therapy. The study involved fabrication of blank and curcumin-loaded graphene quantum dots, their characterizations (TEM, UV-visible and fluorescence emission spectra), cytotoxicity assay, ROS assay and investigation of enhanced antimicrobial photodynamic effect against resistant microbes. The fabrication of blank and loaded graphene quantum dots was confirmed by the observation of peak shift and changes in peak intensity of blank graphene quantum dots, curcumin alone compared with curcumin-loaded graphene quantum dots in UV-visible and fluoresce emission spectra. Cytotoxicity assay showed that 100 µm concentration was not toxic to NIH/3t3 fibroblasts. In ROS assay, the curcumin-loaded formulation showed three-fold increase in ROS production. Blue-light (405 nm) irradiance of 30 J cm -2 and photosensitizer concentration of 100 µm showed ~3.5 log 10 enhanced CUF reduction against Pseudomonas aeruginosa, MRSA, Escherichia coli and Candida albicans. In conclusion, curcumin-loaded graphene quantum dots shoed enhanced antimicrobial photodynamic effects and can be used as an alternative effective treatment for resistant infections., (© 2021 American Society for Photobiology.)

This study was planned to explore the in-vitro and in-vivo therapeutic significance of Paeonia emodi-mediated zinc oxide nanoparticles (ZnO NPs) against the Staphylococcus aureus and Escherichia coli. The texture parameters were derived from nitrogen adsorption-desorption data using Brunauer-Emmett-Teller (BET) and Barrett-Joyner-Halenda (BJH) methods, and the surface area (S BET ) was found to be 214 m 2 /g with a pore size of 2.3 nm. The crystallographic parameters were investigated through X-ray diffraction analysis, and the calculated crystallite size is 29.13 nm. The microstructure was examined through transmission and scanning electron microscopies (TEM and SEM, respectively), and the average particle size estimated from a TEM image is 44.40 nm. The chemical composition and attached function groups were identified through energy-dispersive X-ray and Fourier transform infrared spectroscopies. The in-vitro minimum inhibitory concentration (MIC) for both bacterial species results was found less than 2 μg/ml. The tolerance limit of mouse models was evaluated by the inoculation of different concentrations of ZnO suspension where the concentration above 23 ppm was proved lethal. The maximum infection was caused in mouse models by inoculation of 3 × 10 7 CFUs (Colony forming unit) of the both bacterial species. The concentration higher than 3 × 10 7 CFUs led to the ultimate death of the mice. The histopathological and hematological studies reveal that the after simultaneous inoculation of both ZnO NPs and bacterial suspensions (tolerated amount), no/negligible infection was found in the mice model., (© 2021 Wiley Periodicals LLC.)

Purpose: To determine the effect of zinc deficiency on fecal protein, electrolyte, and short-chain fatty acid levels in both heat-stable (ST) and heat-labile (LT) enterotoxigenic Escherichia coli (ETEC)-induced diarrhea in rats., Methods: Albino rats, weighing 100 to 150 g, were divided into 2 groups, with 15 animals each: non-zinc and zinc-deficient. These two groups were sub-divided into three sub-groups with five rats each: control (saline); LT-ETEC; and ST-ETEC. Sodium phytate (30 mmol/L) was added to the animals' water to induce zinc deficiency, while diarrhea was induced using 5×10 9 ETEC cells/mL. Fecal protein levels were estimated using the Bradford method, while sodium and potassium levels were determined using atomic absorption spectrophotometry. Short-chain fatty acids were measured using gas chromatography-mass spectrometry., Results: Among the non-zinc and zinc-deficient groups, there were significant increases ( p =0.04), ( p =0.03) in fecal protein concentrations (mg/mL) in the LT-ETEC- (4.50±0.33), (6.50±0.26) and ST-ETEC- (3.85±0.19), (5.98±0.32) induced groups compared to the control groups (2.60±0.52), (3.50±0.11) respectively. Fecal sodium and potassium levels (mg/L) were significantly ( p =0.029) increased in non-zinc-deficient rats induced with LT-ETEC (9.35±0.95, 1.05±0.48), and ST-ETEC (9.96±1.02, 1.21±0.45) compared with the control group (8.07±0.44, 0.47±0.17) but the increase were not statistically significant ( p =0.059) in the zinc deficient rat groups. Fecal acetate and propionate levels (mg/g) significantly ( p =0.032) increased when induced with LT-ETEC and ST-ETEC in non-zinc and zinc-deficient groups compared with the control groups., Conclusion: Zinc deficiency among rats with ETEC-induced diarrhea elevated fecal protein loss but may not have an effect on fecal sodium, potassium and short-chain fatty acid levels., Competing Interests: Conflict of Interest: The authors have no financial conflicts of interest., (Copyright © 2022 by The Korean Society of Pediatric Gastroenterology, Hepatology and Nutrition.)

The descriptive cross-sectional study was planned to evaluate drug-related problems, including drug-drug interactions, dose error, use of nephrotoxic drugs and polypharmacy, with special emphasis on kidney disease patients. The study was conducted from January to June 2019 in the Nephrology Ward of Ayub Teaching Hospital, Abbottabad, Pakistan. Doses of medicine and drug-drug interactions were evaluated by comparing it with standard protocols in British National Formulary and Lexicomp. Prescriptions were also evaluated for polypharmacy and use of nephrotoxic drugs. Out of 131 patients, 72 (55%) were males. Drug-drug interactions were found in 69 (52.7%) patients among whom the highest percentage was of moderate drug-drug interaction 63 (48.1%), followed by major 39 (29.8%) and minor 29 (22%) drug-drug interactions. Incidence of polypharmacy 68 (51.9%) and use of nephrotoxic drug 101 (77%) was high, while dose error was low 14 (10.7%). All drug-related problems were present with a high percentage in patients with chronic kidney diseases 29 (78.4) out of 37 (28.2%) such patients. There was significant association of chronic kidney diseases stages with drug-drug interactions, polypharmacy, dose error and prescribing drugs (p<0.05). The higher incidence of drug-related problems reflected irrational prescribing trends and deficiency of professional staff dealing with kidney disease patients.

The Paeonia emodi ( P. emodi )-mediated iron oxide nanoparticles (Fe 2 O 3 NPs) were screened for in-vitro and in-vivo antibacterial activity against the Staphylococcus aureus ( S. aureus ) (ATCC #: 6538) and Escherichia coli ( E. coli ) (ATCC #:15224). The synthesized Fe 2 O 3 NPs were characterized via nitrogen adsorption-desorption process, X-ray diffractometer (XRD), transmission and scanning electron microscopies (TEM and SEM), energy dispersive X-ray (EDX) and Fourier transform infrared (FTIR) spectroscopies. The S BET was found to be 94.65 m 2 /g with pore size of 2.99 nm, whereas the average crystallite and particles size are 23 and 27.64 nm, respectively. The 4 μg/mL is the MIC that inhibits the growth of E. coli, whereas those for S. aureus are below the detection limit (<1.76 μg/mL). The tolerance limit of the mice model was inspected by injecting different concentration of Fe 2 O 3 NPs and bacteria suspensions. The 14 ppm suspension was the tolerated dose and the concentration above were proved lethal. The most severe infection was induced in mice with injection of 3 × 10 7 CFUs of both bacteria, while the inoculation of higher concentrations of bacterial suspensions resulted in the mice's death. The histopathological and hematological studies reveals that the no/negligible infection was found in the mice exposed to the simultaneous inoculation of Fe 2 O 3 NPs (14 ppm) and bacterial suspensions (3 × 10 7 CFUs).

Background: Incoherent use of antibiotics has led toward resistance in MRSA, becoming multidrug-resistant with a high rate of virulence in the community and hospital settings., Objective: Synergistic anti-MRSA activity was investigated in this study for hybrid material composite spheres of amoxicillin, Ag nanoparticles, and chitosan, which were prepared by one-step synthesis method, and various characterizations were performed., Methods: Antimicrobial-susceptibility assay on MRSA was achieved by disc diffusion and agar dilution techniques, while agar well diffusion was used for hybrid composite spheres. The in vitro and cytotoxicity studies were conducted on the skin abrasion mouse model and MTT assay on RD cell, respectively., Results: All isolates showed resistance to the tested antibiotics except vancomycin. MIC against MRSA showed high resistance with amoxicillin from 4 to 128 mg L -1 . The mean diameter of chitosan spheres and Ag nanoparticles was 02 mm and 277 nm, respectively. Morphology of spheres was uneven, varied, porous, and irregular in SEM, and Ag nanoparticles presence and formation was also seen in the micrograph. No substantial interface among drug, nanoparticles, and polymer was found in XRD, and IR showed characteristic peaks of all compounds in the formulation. The in vitro assay showed augmented anti-MRSA activity with amoxicillin loaded hybrid composite spheres (22-29 mm). A significant reduction in microbial burden (~6.5 log 10 CFU mL -1 ) was seen in vivo with loaded hybrid composite spheres formulation. The MTT assay indicated no potential cytotoxicity with hybrid composite spheres., Conclusion: The synergistic effect of Amoxycillin in the current study predicts a promising hybrid formulation with enhanced anti-MRSA activity., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Phragmanthera capitata was reported to possess many biological properties making it a good candidate for the formulation of a phytomedicine with multiple effects. In this work, we studied some factors likely to modify these therapeutic properties with the aim to contribute to its standardization as an improved traditional medicine. P. capitata parasitizing Persea americana, Psidium guajava, and Podocarpus mannii were harvested at three phenological stages (vegetative, flowering, and fruiting stages). The extracts were prepared by maceration in n-hexane, ethyl acetate, ethanol, methanol, and distilled water. The total phenolic, flavonoid, flavonol, and tannin contents were measured using appropriate methods. The antioxidant potential of extracts was investigated using TAC, DPPH scavenging, and FRAP methods. The α -amylase and α -glucosidase inhibitory activities of extracts were determined using enzymatic methods. The ethyl acetate extracts with the best phenolic content were subjected to HPLC analysis. The extraction yields were higher with methanol. The ethyl acetate extract of P . capitata harvested from P. guajava showed a stable HPLC profile during the development of the plant, while extracts from the plant collected from P. americana and P. mannii showed both qualitative and quantitative variations according to phonological stages of the plant. The inhibition of α -amylase was more pronounced for P. capitata harvested from P. guajava , decreasing during flowering and fruiting, while inhibition of α -glucosidase was not influenced by the phenological stage and the host of the plant. The α -amylase inhibitors were better extracted by ethyl acetate and those of α -glucosidase by ethanol or methanol. The phenolic contents and antioxidant properties of the extracts were influenced by the phenological stage of P. capitata and its hosts. These results suggest that it is preferable to harvest P. capitata during flowering or during fruiting stages on any host. None of the used solvents permitted an optimal extraction of active principles form P . capitata , suggesting that the mixture of solvents must be considered in further studies., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2020 Césaire Feudjio et al.)

Background: Biofilm forming ability of Pseudomonas aeruginosa make them vulnerable, because it makes them recalcitrant against various antibiotics. Quorum sensing (QS) is cell density based signaling that helps in bacterial cell-cell communication, which regulated various virulence factors such as pigment and biofilm formation that contribute in the establishment of chronic infections. The interruption of QS is one of the effective approach to control various virulence factors. Present study was intended with the aim to authenticate antibiofilm potential in different solvents based extracts of selected medicinal plant species viz. Berginia ciliata, Clematis grata and Clematis viticella traditionally used by the inhabitants of Himalayan region of Pakistan to treat various pathogenic diseases. P. aeruginosa PAO1, an opportunistic pathogen and involves in various life-threatening infections specifically in immune deficient patients was used as a model pathogen., Methods: Plants were extracted in various organic (ethanol, methanol, acetone, ethyl acetate, hexane, chloroform) as well as in aqueous solvents and their ability to inhibit biofilm was measured. Biofilm of PAO1 was grown in Jensen's medium while growing at 30°C and crystal violet assay was performed to assess the biofilm inhibiting activity of plant extracts., Results: Solvents play a vital role in extraction of plant components and it was found that the plants in various solvents exhibit different activity against the PAO1 biofilm. Comparatively, 1% methanolic extract of B. ciliata (rhizome with skin), showed more than 80% inhibition of biofilm formation without effecting on the growth of the bacterium. Significant correlation between flavonoids content and antibiofilm activity in methanolic extract revealed the contribution of secondary metabolites in P. aeruginosa (PAO1) biofilm inhibition., Conclusion: Our study revealed that plants under investigation more specifically B. ciliata could be a potential candidate for drug discovery to treat P. aeruginosa PAO1, induced infectious diseases especially for its biofilm treatment., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

In this study, molecular imprinted polymer (MIP)-based impedimetric sensor has been developed to detect dengue infection at an early stage. Screen-printed carbon electrode (SPCE) was modified with electrospun nanofibers of polysulfone (PS) and then, coated with dopamine while using NS1 (non-structural protein 1-a specific and sensitive biomarker for dengue virus infection) as template during polymerization. The self-polymerization of dopamine at room temperature helps to retain exact structure of template (NS1) which results in generating geometrically fit imprinted sites for specific detection of target analyte. The electrochemical properties of MIP-modified SPCEs were studied by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) at every step of modification. Under optimal conditions, impedimetric measurements showed linear response in the range from 1 to 200 ng/mL. The developed sensor can selectively detect NS1 concentrations as low as 0.3 ng/mL. Moreover, impedimetric sensor system was also employed for NS1 determination in real human serum samples and satisfying recoveries varying from 95 to 97.14% were obtained with standard deviations of less than 5%.

The current study is an attempt to explore the effect of varying quantities of hydroxypropyl cellulose (HPC) polymer on carbamazepine (CBZ) cocrystal formation with dicarboxylic acid coformers i.e., malonic acid (MA), succinic acid (SA), glutaric acid (GA), and adipic acid (AA). The cocrystals were first prepared without polymer by slurry crystallization method and then tried with different quantities of the polymer. The prepared samples were characterized by Powder X-ray Diffraction (XRPD). The characterization results indicate that in methanol pure carbamazepine-malonic (CBZ-MA) and carbamazepine-adipic acid (CBZ-AA) cocrystal can be prepared, while in ethanol and acetone pure carbamazepine-succinic (CBZ-SA) and carbamazepine-glutaric acid (CBZ-GA) cocrystals can be obtained respectively. The same cocrystals were tried using HPC polymer in three different quantities. The characterization results showed that a higher quantity of HPC polymer transforms CBZ-MA cocrystal polymorph-I to polymorph-II. The CBZ-SA and CBZ-GA cocrystal formation somehow inhibited as the concentration of HPC polymer increases. But on the other side, the formation of CBZ-AA cocrystal utterly not inhibited in the presence of varying quantities of HPC polymer. Furthermore, 11 different quantities of HPC were tried to know about the inhibitory concentration of HPC on CBZ-AA cocrystal formation. The CBZ-AA cocrystal preparation was not inhibited even at higher quantities of HPC compared to the coformer. Additionally, the effect of three different quantities of HPC on the thermal stability of the CBZ-AA cocrystal was investigated. Moreover, the stability of pure CBZ at 92% relative humidity (RH) condition was compared to CBZ-AA cocrystal with and without HPC polymer. The CBZ-AA cocrystal with and without HPC polymer was more stable than pure CBZ.

Biogenic production of nanoparticles is eco-friendly, less expensive method with various medical and biological applications. Nanotechnology along with photodynamic therapy is gaining tremendous importance with enhanced efficacy. The present work was aimed to evaluate methanolic extracts and nanoparticles of two selected plants (Datura suavolens and Verbina tenuisecta) for cytotoxic photodynamic, antioxidant and antimicrobial study. Both extract and silver (5 mM) nanoparticles of Datura plant showed significant activities against bacterial strains. Maximum ZOI of 27.3 ± 1.6 mm was observed with nanoparticles of Datura branches with minimum inhibitory (MIC) value of 32 µg/ml. In case of antifungal and antioxidant assay samples were moderately active. Silver nanoparticles and extracts were effective against rhabdomyosarcoma cell line with lowest IC 50 value of 42.5 ± 0.6 μg/ml and percent viability of 25.6 ± 1.3 of Verbena tenuisecta. However, nanoparticles of Datura leaves and branches were more potent with IC 50 value of 2.4 ± 0.9 μg/ml and 7.8 ± 1.1 μg/ml respectively. The result of photodynamic study showed that efficacy of photosensitizer was enhanced and percent viability reduced when nanoparticles used as an adjunct. The color change and UV spectra (415‒425 nm) indicated the production of nanoparticles. Fourier transform infrared spectroscopy (FTIR) spectra showed presence of different functional groups e.g., hydroxyl, carbonyl and amino. Nanoparticles are sphenoid in morphology and size ranges between 20-150 nm. Current study showed these silver nanoparticles can be used as cytotoxic agent in photodynamic therapy and can play a critical role to establish medicinal potential of selected plants.

Background: Staphylococcus aureus (S. aureus) is the most common infectious agent in the community and hospitals. Infections with S. aureus are now becoming difficult to be treated by using conventional antibiotics due to its emerging methicillin-resistant S. aureus (MRSA) strain., Objective: In the present study, MRSA was isolated from clinical samples and evaluated for resistance against different antibiotics, TiO2 nanoparticles, and their combinations., Methods: Clinical samples were collected from Ayub Medical Complex (AMC), Abbottabad, Pakistan, and identified by different biochemical tests and polymerase chain reactions (PCR). Kirby-Bauer disk diffusion method was performed to evaluate antimicrobial susceptibility. Minimum Inhibitory Concentration (MIC) of ampicillin, ciprofloxacin, erythromycin, and vancomycin was found out by agar dilution method while the broth dilution method was used for the MIC of TiO2 nanoparticles and their combinations with erythromycin., Results: All 13/100 (13%) MRSA were successfully identified. All isolates were susceptible to quinupristin/ dalfopristin, teicoplanin, and vancomycin, while the highest resistance was seen with erythromycin, penicillin, and tetracycline. MIC showed high resistance against ampicillin (0.25-512 mg/L) and erythromycin (0.25-1024 mg/L)., Conclusion: The MIC value of 2 mM TiO2 nanoparticles was found to be the most effective concentration after 12 h of incubation, while the combination of erythromycin with 3 mM TiO2 nanoparticles was found to be more potent which significantly lowered down the MIC of erythromycin to 2-16 mg/L., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Aim of this study was to synthesize new inhibitors on the basis of active site of aspartic protease enzyme and to evaluate their intended biological activity. A3D model of an enzyme was generated via homology modeling and series of novel amide ligands were synthesized by using a short high yield process, subsequently, analyzed in-silico and in-vitro anti-leishmanial activities. Characterization and identification was accomplished via NMR (H 1 & C 13 ), infrared and mass spectroscopic techniques. Among all compound (4) was found to show significant activity (IC 50 58±0.01) against Leishmania major (L. major) species. Furthermore, docking studies confirmed the inhibition of a targeted enzyme that supported the interaction of potent compound (4) with key residues (aspartic protease) via hydrogen bonds. Present study conferred about novel compound (4) as a promising compound to antagonize L. major activities in future.

Medicinal plants of Pakistan are known for their curative properties against snake bite as rural people have been using natural herbs for such injuries for hundreds to thousands of years. People of rural areas of Pakistan are prone to snakebite, and on the whole death due to snakebite has been increasing worldwide. The objective of this study was to test the neutralizing potential of 17 Pakistani medicinal plant extracts against phospholipase A2 activity in Echis carinatus venom. Plant material was extracted by simple maceration and fractionation of active plant extracts. Venom was collected by manual massage of the venom glands. The PLA2 enzymatic assay was performed to map out the venomous activity of Echis carinatus envenomation. Snake venom released fatty acids at different concentrations (0.1-5 mg/ml) of venom in a dose-dependent manner. Reduction of pH by 01 correlated with 133 μmol of fatty acids released at 5mg/ml of venom. All plants extract inhibited PLA2 activity, however, Curcuma longa, Citrullus colocynthis and Rubia cordifolia inhibited maximum of PLA2 activity (⁓78%) comparable to the standard antidote (p>0.5). Medicinal plants possess secondary metabolites and many active compounds that may have neutralizing or inhibiting properties against the PLA2 activity of Echis venom. Further studies such as compound analysis could provide an alternative against snakebites injuries resulting from Echis carinatus venom.

Objective: To assess the knowledge, attitude and practice of hepatitis B and C patients towards the disease., Methods: The cross-sectional, descriptive, questionnaire-based study was conducted at the Rawalpindi Medical University (RMU) and its allied hospitals, Rawalpindi, Pakistan, from October to December 2017, and comprised hepatitis B and C patients aged ≥18 years who were visiting the outpatient department. A structured, close-ended, 30-item questionnaire was used to collect data which was then analysed using SPSS 23. ., Results: Of the 399 patients, 230(57.6%) were women, 103(26%) did not have any formal education, and TV, radio and the internet was the main source of information regarding hepatitis for 138(34.6%) patients. The overall mean age was 42.82}11.78 years. The sample showed poor knowledge with a mean score of 7.33}2.03, adequate attitude with a mean of 4.33}1.19, and poor practice with a mean of 2.97}1.05. Correlation was statistically significant between knowledge and practice (p<0.01)., Conclusions: Lack of imperative knowledge of hepatitis in patients was the main reason for negative attitude and poor practices.

Background Distemonanthus benthamianus is used in the Western part of Cameroon to treat diarrheal episodes and infections. This study assessed its trunk-bark extracts activity against enteropathogenic Escherichia coli 31 (EPEC 31)-induced diarrhea in rats. Methods Aqueous and methanolic extracts were analyzed through high-performance liquid chromatography (HPLC). In vitro minimum inhibitory and bactericidal concentrations (MICs/MBCs) were evaluated on Enterococcus faecalis (ATCC 10,541), E. coli (ATCC 6539), Klebsiella pneumoniae (ATCC 13,883), Salmonella typhi (ATCC 6539) strains and on Proteus mirabilis, Pseudomonas aeruginosa (PA 01) and Shigella flexneri isolates using the microdilution method. Diarrhea was induced by inoculating rats with EPEC 31 (1.5 × 108 CFU/mL; p.o). Serum transaminases level assay and enzyme-linked immunosorbent assay (ELISA) for cytokines determination were performed. Hematoxylin-eosin (H-E) staining was used for intestinal tissue analysis. Results HPLC fingerprints of extracts showed presence of gallic acid and other unidentified compounds. The lowest MIC of 256 µg/mL was obtained with methanolic extract. At 100 mg/kg, both extracts significantly (p<0.001) inhibited diarrhea, with the methanolic extract being the most active. In addition, the methanolic extract significantly (p<0.001) increased the relative mass of the liver compared to negative control (Tween-DMSO 8%). The aqueous extract (100 mg/kg) significantly (p<0.01) increased alanine aminotransferase (ALT) serum concentration; while the methanolic extract (100 mg/kg) exhibited similar effect over aspartate aminotransferase (AST). At 50 and 100 mg/kg, the methanolic extract significantly (p<0.05 and p<0.01) decreased the Interleukin-1β (IL-1β) serum level, compared to negative control (Tween-DMSO 8%). Serum level of tumor necrosis factor alpha (TNF-α) significantly (p<0.001) decreased with 100 mg/kg of aqueous extract and all doses of methanolic extract. Inhibition of inflammatory cells tissue infiltration and epithelial regeneration was highly noticed in the ileum and colon of extracts-treated rats than in ciprofloxacin-treated animals. Conclusion These findings suggest that D. benthamianus trunk-bark extracts displayed therapeutic effects against infectious diarrhea in rats.

This article reports moxifloxacin (Mox)-loaded nanocomposite films (CSN) of chitosan and chemically reduced silver (Ag). The synthesis of silver nanoparticles was confirmed by specific surface plasmon resonance (SPR) peaks detected via UV-Visible spectroscopy at the wavelength range of 400-450 nm. The embedded Mox was chemically characterized and kinetically analyzed for in-vitro drug release and ex-vivo drug permeation through rat skin. The prepared films presented higher swelling ratio and lower tensile strength (TS) and better elongation at break (EB) than control formulation (pure chitosan film). All the prepared Mox-loaded, non-crosslinked formulations presented sustained release of drug up to 12 h while slow and prolonged drug release up to 36 h was observed in Mox-loaded crosslinked CSN films. Drug permeation studies indicated that the maximum cumulative amount of Mox permeated (%) among Mox-loaded, non-crosslinked CSN films was displayed by CSM1 (57.79%); while in case of Mox-loaded, crosslinked CSN films, the highest drug permeation was presented by CSM18 (62.87%) in 24 h. The antibacterial efficacy of the prepared films was tested in-vitro against S. aureus (ATCC # 6538), P. aeruginosa (ATCC # 9721) and two clinically isolated strains of methicillin resistant S. aureus (MRSA). CSN films presented excellent against the all the selected strains with antibacterial potential being highest against S. aureus. In summary, the promising antibacterial potential of the CSN films recommend its biomedical application for use in wound dressing., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: To evaluate the knowledge, attitude and practice of self-medication in medical undergraduate students., Methods: This cross-sectional questionnaire-based survey was conducted from January to June, 2017, among undergraduate students of Ayub Teaching Hospital, Women Medical College, International Medical College and Frontier Medical College in Abbottabad, Pakistan. The sample comprised students of first, second, third, fourth and fifth professional year. Data was collected using self-generated questionnaire., Results: Of the 400 subjects approached, 300(75%) returned the questionnaire fully completed. Of them, 208 (69.3%) were females and 92(30.66%) were males. The most common age group was 22-25 years 182(60.7%). Self-medication was found to be prevalent among 297(99%) subjects and 139(46.3%) respondents had practised self-medication in the preceding 6 months more than two times. Over-the-counter drugs were commonly used for the self-medication in 295(98.3%) cases. Overall, 285(95%) respondents had a positive attitude towards self-medication. Media was found to be the most common source of information for 93(31%) cases., Conclusions: High prevalence of self-medication was noticed, with over-the-counter drugs being the most commonly used. Proper educational awareness programmes about self-medication can control the issue.

Purpose: In the present study, enhancement of the the antibacterial activity of ceftriaxone against Gram-positive (meticillin-resistant Staphylococcus aureus; MRSA) and Gram-negative (Escherichia coli) bacteria with a biodegradable polymer was attempted., Methodology: MRSA and E. coli were collected and identified by biochemical and molecular tests. Blank and ceftriaxone-loaded chitosan nanoparticles (CNPs) were prepared by the ionic gelation method. In vitro antibiotic-susceptibility studies were performed by disc diffusion, agar well plate method, Etest and time-kill assay. In vivo activity was assessed using the neutropenic mouse thigh model and cytotoxicity was estimated by MTT (methylthiazolyldiphenyl tetrazolium bromide) assay with the MCF-7 cancer cell line., Results: MRSA showed 97 % and E. coli 83 % resistance against ceftriaxone in the disc diffusion test. The isolates showing a ≥1024 mg l-1 MIC value for ceftriaxone were selected for further evaluation. In the agar well plate method, the mean zones of inhibition for blank and ceftriaxone-loaded CNPs were 17 and 23 mm, respectively, for MRSA isolates and 15 and 25 mm, respectively, for E. coli isolates. In the time-kill assay, ~1 log10 to ~2.5 log10 reduction in viability was seen with both isolates when treated with ceftriaxone-loaded CNPs over 24 h. The in vivo studies also showed the enhanced antibacterial activity of ceftriaxone-loaded CNPs, with a 41 % reduction in MRSA and a 27 % reduction in E. coli burden. A low cytotoxicity of blank and ceftriaxone-loaded CNPs was seen, with a slight reduction in the percentage viability of cells from 87 to 83 % and from 88 to 81 %, respectively., Conclusion: The synergistic effect of ceftriaxone-loaded CNPs is a useful finding for the treatment of MRSA and E. coli infections.

Objective: To investigate the efficacy and impact of Protection Against Harassment of Women at Workplace Act 2010 in the public health sector in its socio-legal perspective., Methods: This cross-sectional study was conducted from July to December 2014 in Abbottabad, Pakistan, and comprised subjects selected from 53 basic health units managed by the government. SPSS 20 was used for statistical analysis., Results: Of the 450 questionnaires, 430(96.6%) were returned duly filled. Overall, 40% male (120 respondents) and 26% female (34 respondents) knew about the Act. Besides, 39% males (117 respondents) and 63% females (82 respondents) appeared unsatisfied with the complaint mechanism prescribed in the Act; all the respondents established that no case had been registered after the introduction of the Act. Overall, 31% males (93 respondents) and 57% females (74 respondents) thought that without the health governance commitment and social support structure the Act could not work., Conclusions: The Protection Against Harassment of Women at Workplace Act 2010 was considered ineffective by the respondents who were concerned about the lack of social support system for the Act.

Abstract: The objective of this study was to fabricate topical formulations of diclofenac diethylamine (DD) using isopropyl myristate (IPM) and isopropyl palmitate (IPP) as permeation enhancers. Franz cell and bacterial cellulose were used as analytical instrument and diffusion membrane, respectively. Permeation enhancers exhibited significant effect on the permeation characteristics of DD. It was concluded from the results that improved permeation of DD was observed when IPP was used as enhancer.

Background: Enterococci normally inhabit the intestinal tract of humans and are also a potential pathogen in causing nosocomial infections. The increase in antibiotic resistance and transfer of antibiotic resistance gene to Staphylococcus aureus (S. aureus) due to co-colonization has increased its importance in research. The aim of the study was to evaluate local epidemiology of nasal and rectal colonization with Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) in patients of Paediatrics Intensive Care Unit (PICU) and correlation with clinical and socioeconomic factors., Methods: The nasal and perirectal swab samples were collected from 110 patients admitted in PICUs of three tertiary care hospitals of Rawalpindi Medical College, Pakistan. The identification of enterococci was done by biochemical tests and by PCR for ddl, vanA and vanB genes. Antibiotic susceptibility testing was performed by disc diffusion and MICs were determined for vancomycin, tetracycline, ciprofloxacin and oxacillin only., Results: Out of 220 nasal and perirectal samples, 09 vancomycin-resistant enterococci (VRE) and 76 vancomycin-susceptible enterococci (VSE), consisting of 40 E. faecalis and 45 E. faecium were isolated. PCR successfully identified both species with ddl primers and VRE with vanA primer. With disc diffusion method, all isolates were resistant to most of the antibiotics tested except linezolid, quinupristin/dalfopristin, teicoplanin and vancomycin. VRE showed resistance to teicoplanin and vancomycin both and none was resistant to linezolid and quinupristin/dalfopristin. Generally, E. faecium isolates were more resistant than E. faecalis. MICs of vancomycin for nasal and perirectal VRE were 512 mg/L and 64 to 512 mg/L respectively. VRE were more in patients with prolonged hospitalization, from urban localities and those having pneumonia., Conclusion: Present study reveals high colonization and antibiotic resistance in enterococcal isolates from nasal and perirectal area. Nasal colonization by enterococci in PICU is more alarming as VRE may cause infection and can transfer this resistance gene to other microorganisms like S. aureus.