Cocrystals of a coumarin derivative: an efficient approach towards anti-leishmanial cocrystals against MIL-resistant Leishmania tropica.

Leishmaniasis is a neglected parasitic tropical disease with numerous clinical manifestations. One of the causative agents of cutaneous leishmaniasis (CL) is Leishmania tropica (L. tropica) known for causing ulcerative lesions on the skin. The adverse effects of the recommended available drugs, such as amphotericin B and pentavalent antimonial, and the emergence of drug resistance in parasites, mean the search for new safe and effective anti-leishmanial agents is crucial. Miltefosine (MIL) was the first recommended oral medication, but its use is now limited because of the rapid emergence of resistance. Pharmaceutical cocrystallization is an effective method to improve the physicochemical and biological properties of active pharmaceutical ingredients (APIs). Herein, we describe the cocrystallization of coumarin-3-carboxylic acid (CU, 1a; 2-oxobenzopyrane-3-carboxylic acid, C10H6O4) with five coformers [2-amino-3-bromopyridine (1b), 2-amino-5-(trifluoromethyl)-pyridine (1c), 2-amino-6-methylpyridine (1d), p-aminobenzoic acid (1e) and amitrole (1f)] in a 1:1 stoichiometric ratio via the neat grinding method. The cocrystals 2-6 obtained were characterized via single-crystal X-ray diffraction, powder X-ray diffraction, differential scanning calorimetry and thermogravimetric analysis, as well as Fourier transform infrared spectroscopy. Non-covalent interactions, such as van der Waals, hydrogen bonding, C-H...π and π...π interactions contribute significantly towards the packing of a crystal structure and alter the physicochemical and biological activity of CU. In this research, newly synthesized cocrystals were evaluated for their anti-leishmanial activity against the MIL-resistant L. tropica and cytotoxicity against the 3T3 (normal fibroblast) cell line. Among the non-cytotoxic cocrystals synthesized (2-6), CU:1b (2, IC50 = 61.83 ± 0.59 µM), CU:1c (3, 125.7 ± 1.15 µM) and CU:1d (4, 48.71 ± 0.75 µM) appeared to be potent anti-leishmanial agents and showed several-fold more anti-leishmanial potential than the tested standard drug (MIL, IC50 = 169.55 ± 0.078 µM). The results indicate that cocrystals 2-4 are promising anti-leishmanial agents which require further exploration.

Study of drug resistance-associated genetic mutations, and phylo-genetic analysis of HCV in the Province of Sindh, Pakistan.

Current management of HCV infection is based on Direct-Acting Antiviral Drugs (DAAs). However, resistance-associated mutations, especially in the NS3 and NS5B regions are gradually decreasing the efficacy of DAAs. The aim of the current study was to identify such mutations in the NS3, and NS5B genes in DAAs treatment-naïve Pakistani chronic HCV 3a patients. Peripheral blood samples were collected from 233 chronic HCV 3a patients at different tertiary care hospitals in Karachi, Pakistan, between August 2020 to September 2021. PCR-amplified target regions of the NS3/NS5B gene were subjected to Sanger sequencing to identify resistance-associated mutations. Phylogenetic analysis of the identified amino acid sequences was performed using HCV3a sequences of the global population in the virus pathogen resource (VIPR) database. Sequence analysis identified five amino acid mutations, Leu36Pro, Gln41His, Gln80Lys/Arg, Ala156Tyr, and Gln168Arg in the NS3 region, and two mutations Leu159Phe and Cys316Arg in the NS5B region. Phylogenetic analysis revealed a high genetic diversity in the studied isolates. Overall, the prevalence of resistance-associated substitutions was almost similar to other geographic regions worldwide. This data could be helpful in selecting the most effective treatment regimen for HCV chronically infected people in Pakistan.

New cocrystals of heterocyclic drugs: structural, antileishmanial, larvicidal and urease inhibition studies.

Many heterocycles have been developed as drugs due to their capacity to interact productively with biological systems. The present study aimed to synthesize cocrystals of the heterocyclic antitubercular agent pyrazinamide (PYZ, 1, BCS III) and the commercially available anticonvulsant drug carbamazepine (CBZ, 2, BCS class II) to study the effect of cocrystallization on the stability and biological activities of these drugs. Two new cocrystals, namely, pyrazinamide-homophthalic acid (1/1) (PYZ:HMA, 3) and carbamazepine-5-chlorosalicylic acid (1/1) (CBZ:5-SA, 4), were synthesized. The single-crystal X-ray diffraction-based structure of carbamazepine-trans-cinnamic acid (1/1) (CBZ:TCA, 5) was also studied for the first time, along with the known cocrystal carbamazepine-nicotinamide (1/1) (CBZ:NA, 6). From a combination drug perspective, these are interesting pharmaceutical cocrystals to overcome the known side effects of PYZ (1) therapy, and the poor biopharmaceutical properties of CBZ (2). The purity and homogeneity of all the synthesized cocrystals were confirmed by single-crystal X-ray diffraction, powder X-ray diffraction and FT-IR analysis, followed by thermal stability studies based on differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). Detailed intermolecular interactions and the role of hydrogen bonding towards crystal stability were evaluated quantitatively via Hirshfeld surface analysis. The solubility of CBZ at pH 6.8 and 7.4 in 0.1 N HCl and H2O were compared with the values of cocrystal CBZ:5-SA (4). The solubility of CBZ:5-SA was found to be significantly improved at pH 6.8 and 7.4 in H2O. All the synthesized cocrystals 3-6 exhibited a potent urease inhibition (IC50 values range from 17.32 ±â€…0.89 to 12.3 ±â€…0.8 µM), several times more potent than standard acetohydroxamic acid (IC50 = 20.34 ±â€…0.43 µM). PYZ:HMA (3) also exhibited potent larvicidal activity against Aedes aegypti. Among the synthesized cocrystals, PYZ:HMA (3) and CBZ:TCA (5) were found to possess antileishmanial activity against the miltefosine-induced resistant strain of Leishmania major, with IC50 values of 111.98 ±â€…0.99 and 111.90 ±â€…1.44 µM, respectively, in comparison with miltefosine (IC50 = 169.55 ±â€…0.20 µM).

Repurposing of US-FDA approved drugs against SARS-CoV-2 main protease (Mpro) by using STD-NMR spectroscopy, in silico studies and antiviral assays.

SARS-CoV-2 Main protease (Mpro) is a well-known drug target against SARS-CoV-2 infection. Identification of Mpro inhibitors is vigorously pursued due to its crucial role in viral replication. The present study was aimed to identify Mpro inhibitors via repurposing of US-FDA approved drugs by STD-NMR spectroscopy. In this study, 156 drugs and natural compounds were evaluated against Mpro. Among them, 10 drugs were found to be interacting with Mpro, including diltiazem HCl (1), mefenamic acid (2), losartan potassium (3), mexiletine HCl (4), glaucine HBr (5), trimebutine maleate (6), flurbiprofen (7), amantadine HCl (8), dextromethorphan (9), and lobeline HCl (10) in STD-NMR spectroscopy. Their interactions were compared with three standards (Repurposed anti-viral drugs), dexamethasone, chloroquine phosphate, and remdesivir. Thermal stability of Mpro and dissociation constant (Kd) of six interacting drugs were also determined using DSF. RMSD plots in MD simulation studies showed the formation of stable protein-ligand complexes. They were further examined for their antiviral activity by plaque reduction assay against SARS-CoV-2, which showed 55-100% reduction in viral plaques. This study demonstrates the importance of drug repurposing against emerging and neglected diseases. This study also exhibits successful application of STD-NMR spectroscopy combined with plaque reduction assay in rapid identification of potential anti-viral agents.

COVID-19 independently predicts poor outcomes in Acute Ischemic Stroke- Insights from a multicenter study from Pakistan and United Arab Emirates.

BACKGROUND: Ischemic stroke associated with coronavirus 2019 (COVID-19) has been well recognized by now. Few studies have compared COVID related versus unrelated strokes. We intend to report on a large group of Asian patients from two countries and compare COVID with non-COVID strokes admitted during the same time period. METHODS: Consecutive cases of acute ischemic stroke either presenting or developing, between March 2020 and December 2021 in four tertiary care hospitals (1 in Dubai, UAE and 3 in Karachi, Pakistan) and testing positive for COVID-19 were included in the study. Patients admitted with ischemic stroke during the same time period and who tested negative for COVID-19 were also randomly selected from the four hospitals. All data was collected from the medical records of the patients and recorded on a standard questionnaire before it was entered in SPSS version 21 for analysis. RESULTS: There were 139 COVID positive and 271 COVID negative patients with acute ischemic stroke included in the current study. There were significantly more males (80.6% vs 64.9%, p=0.001) and more large vessel strokes in the COVID positive group (41% vs 21.8%, p<0.001). Being COVID positive was an independent predictor of poor outcome at discharge, defined as a modified Rankin score of 3-6 (OR 3.87, 95% CI 2.21-6.77) after adjusting for country, age, sex, vascular comorbid conditions and stroke subtype. CONCLUSIONS: In this largest series of patients with COVID related strokes from Asia, COVID-19 was an independent predictor of poor outcomes at discharge after adjusting for other variables.

Anti-leishmanial physalins-Phytochemical investigation, in vitro evaluation against clinical and MIL-resistant L. tropica strains and in silico studies.

Cutaneous leishmaniasis (CL) is a major health problem in over 98 countries of the world, including Pakistan. The current treatments are associated with a number of adverse effects and availability problem of drugs. Therefore, there is an urgent need of easily available and cost effective treatments of CL- in Pakistan. The bioassay-guided fractionation and purification of crude extract of Physalis minima has led to the isolation of a new aminophysalin B (1), and eight known physalins, physalin B (2), 5ß,6ß-epoxyphysalin B (3), 5α-ethoxy-6ß-hydroxy-5,6-dihydrophysalin B (4), physalin H (5), 5ß,6ß-epoxyphysalin C (6), and physalin G (7), K (8), and D (9). It is worth noting that compound 1 is the second member of aminophysalin series, whereas compound 6 was fully characterized for the first time. The structures of compounds 1-9 were elucidated by spectroscopic techniques Whereas, the structural assignments of compounds 1 and 8 were also supported by single-crystal X-ray diffraction studies. The anti-leishmanial activity of isolated physlains 1-9 was evaluated against Leishmania major and Leishmania tropica promastigotes. Compounds 2, 3, and 5-7 (IC50 = 9.59 ± 0.27-23.76 ± 1.10 µM) showed several-fold more potent activity against L. tropca than tested drug miltefosine (IC50 = 42.75 ± 1.03 µm) and pentamidine (IC50 = 27.20 ± 0.01 µM). Whereas compounds 2, 3 and 5 (IC50 = 3.04 ± 1.12-3.76 ± 0.85 µM) were found to be potent anti-leishmanial agents against L. major, several fold more active than tested standard miltefosine (IC50 = 25.55 ± 1.03 µM) and pentamidine (IC50 = 27.20 ± 0.015 µM). Compounds 4 (IC50 = 74.65 ± 0.81 µM) and 7 (IC50 = 39.44 ± 0.65 µM) also showed potent anti-leishmanial ativity against the miltefosine-unresponsive L. tropica strain (MIL resistant) (miltefosine IC50 = 169.55 ± 0.78 µM). Molecular docking and predictive binding studies indicated that these inhibitors may act via targeting important enzymes of various metabolic pathways of the parasites.

Identification of quinoline derivatives as growth inhibitors of MDR pathogen Klebsiella pneumoniae.

Aims: This study was aimed to identify compounds with significant inhibitory potential against multidrug-resistant (MDR), multidrug-sensitive and clinical isolates of Klebsiella pneumoniae. Materials & methods: Antibacterial activity of the nitroquinoline derivatives was assessed by micro-plate Alamar Blue assay. Results: Nitroquinoline derivatives 9, 11 and 14 showed inhibitory activity against MDR K. pneumoniae. Docking studies of these compounds with topoisomerase IV of K. pneumonia indicated the interactions of these compounds at the active site residues of enzyme near to cofactor (Mg+2). Furthermore, compound 11 was identified as a reactive oxygen species (ROS) inducer. None of the compounds showed hemolytic effect. Conclusion: This study was designed to identify compounds active against MDR K. pneumoniae which causes infections, such as pneumonia and urinary tract infections.

Genome sequencing and analysis of genomic diversity in the locally transmitted SARS-CoV-2 in Pakistan.

Surveillance of genetic diversity of the SARS-CoV-2 is extremely important to detect the emergence of more infectious and deadly strains of the virus. In this study, we evaluated mutational events in the SARS-CoV-2 genomes through whole genome sequencing. The samples were collected from COVID-19 patients in different major cities of Pakistan during the four waves of the pandemic (May 2020 to July 2021) and subjected to whole genome sequencing. Using in silico and machine learning tools, the viral mutational events were analyzed, and variants of concern and of interest were identified during each of the four waves. The overall mutation frequency (mutations per genome) increased during the course of the pandemic from 12.19 to 23.63, 31.03, and 41.22 in the first, second, third, and fourth waves, respectively. We determined that the viral strains rose to higher frequencies in local transmission. The first wave had three most common strains B.1.36, B.1.160, and B.1.255, the second wave comprised B.1.36 and B.1.247 strains, the third wave had B.1.1.7 (Alpha variant) and B.1.36 strains, and the fourth waves comprised B.1.617.2 (Delta). Intriguingly, the B.1.36 variants were found in all the waves of the infection indicating their survival fitness. Through phylogenetic analysis, the probable routes of transmission of various strains in the country were determined. Collectively, our study provided an insight into the evolution of SARS-CoV-2 lineages in the spatiotemporal local transmission during different waves of the pandemic, which aided the state institutions in implementing adequate preventive measures.

16S metagenomics dataset of Zea mays and Triticum aestivum rhizosphere from Kallar Syedan Punjab, Pakistan.

Plant microbiome referred to as plant second genome, plays pivotal role in determination of vigor and productivity of plant. Current high-throughput sequence technologies provide remarkable insight into microbial diversity and host microbe interaction. The obtained dataset aimed to reveal the core bacterial community residing the rhizosphere of two leading cereal crops Zea mays and Triticum aestivum grown in different seasons at the same geographical area. The rhizosphere bacterial communities were explored via amplicon sequencing of V3-V4 region of 16S rRNA region using IonS5™XL sequencing platform. The classified tags for 16S rRNA from both the samples were clustered into 1502 Microbial operational taxonomic units (OTUs) at 97% similarity with 1340 OTUs in Zea mays and 1337 OTUs in Triticum aestivum. Ten bacterial phyla predominant in the rhizosphere were Proteobacteria, Actinobacteria, Firmicutes, Acidobacteria, Bacteroidetes, Chloroflexi, Gemmatimonadetes, Verrucomicrobia, Nitrospirae and Thermomicrobia. These bacterial phyla accounted for 98% and 98.9% of the OTUs in Zea mays and Triticum aestivum, respectively. Statistical analysis depicted the presence of slight variations in the relative abundance of bacterial groups residing the rhizosphere of Zea mays and Triticum aestivum. The community data produced in the present work can be used for meta-analysis studies to understand rhizosphere bacterial community of two major cereal crops. Furthermore, bacterial composition and diversity data is prerequisite for rhizosphere engineering to enhance cereal production to cope with upcoming global challenges of climate change and population growth.

Rice straw biochar in combination with farmyard manure mitigates bromoxynil toxicity in wheat (Triticum aestivum L.).

Herbicide residues in agriculture commodities are a serious threat for human health and crop production. We investigated the efficient utilization of rice straw biochar and farmyard manure as organic amendments in mitigating the bromoxynil residues in leaves, grain, husk and root tissues of wheat plants. Growth and yield of wheat plants were also examined under field conditions through a 02-year field experiment. The experiment was set up using two wheat cultivars (Faisalabad-08 and Galaxy-2013) as main plot factors and different formulations of biochar and farmyard manure as sub-plot factors in a randomized complete Block design (split plot arrangement) with 03 replications. Different formulations of biochar (BC) and farmyard manure (FYM) i.e., 100 BC: 00 FYM, 75 BC: 25 FYM, 50 BC: 50 FYM, 25 BC: 75 FYM, 00 BC: 100 FYM and 00 BC: 00 FYM as control were prepared and mixed with soil (2% m/m) at the time of sowing. The wheat crop was sprayed with recommended dose (Buctril Super 60 EC, 825 mL ha-1) of bromoxynil 40 days after sowing of the crop. Grain, husk, and root samples were collected at maturity while leave samples were taken 10 days after the herbicide application. Results revealed that the organic amendments significantly reduced the bromoxynil concentration in different tissues of wheat plant besides increasing the growth and yield of plants. The highest concentration of bromoxynil residues was found in control treatment whereas; sole biochar (100 BC:00 FYM) reduced the herbicide residues up to 78% in grain and husk of wheat, 40% in leaves and 64% in root tissues along with 41-44% increase in the grain yield. So, rice straw biochar along with farmyard manure could be considered as a promising option for mitigating the residual herbicide issues in the crop plants along with increase in the yield of wheat crop.

Biosynthesis of silver nanoparticles from Mukia maderaspatana (L) and their biological activities.

Nanotechnology is a field of science that consists of atoms, molecules and supramolecular molecules that create nanoparticles ranging in size from 1-100nm. Silver nanoparticles are widely used that are considered as effective antimicrobial agents. In this paper, the antioxidant activity of biosynthesized SNPs were analyzed by the DPPPH activity, hydrogen peroxide activity, hydroxyl RSA, TAC, TFC; their results confirmed that the phenolic compounds of this plant peels extracts enhanced the antioxidant and antiglycation activity with respect to silver nanoparticles. Biosynthesized nanoparticles of this plant extracts also showed strong zone of inhibition against the different Xanthomas, Pseudomonas and E. coli. This study concluded that biosynthesized nanoparticles of Mukia maderaspatna (M.M) plant peels extracts have the great biological activities i.e. antiglycation, antioxidant and antibacterial. More research is needed to know the exact dose rate and to compare the different dose combination of the plant with the strong antibiotic agents against these bacteria.

Frequency of Cerebral Venous Thrombosis in Dubai, UAE-A Multicenter, Retrospective Study.

Background and Purpose: Cerebral venous thrombosis (CVT), also known as cerebral dural venous thrombosis, is not a very common cause of stroke. It has a diverse clinical presentation, with multiple predisposing factors and imaging findings. The incidence of CVT varies in different studies. We hypothesized that the incidence of CVT in Dubai is more than that reported in the West owing to dehydration caused by the high atmospheric temperature. Methods: We retrospectively reviewed the medical records of all patients aged more than 13 years with appropriate International Classification of Diseases, 9th Revision, codes for the diagnosis of CVT from January 1, 2010 to December 31, 2018 from three major hospitals with an acute stroke service in Dubai, United Arab Emirates. Results: We identified 138 patients who presented with CVT. The average frequency of CVT was 6.6 per 100,000 population. Fifty percent of the patients presented between May and September, which are considered the hottest months in the UAE. Headache was the most common presentation, followed by seizures. The most important risk factors were the use of oral contraceptive pills, anemia, and polycythemia. Our study showed a higher incidence of anemia, polycythemia, thrombophilia, and abnormal CT brain findings. Conclusion: In our study, CVT was more common during the months of summer. Anemia and polycythemia were strongly associated with CVT. Public awareness about CVT, its higher incidence during summer months and strategies to avoid dehydration might be considered to reduce the incidence of CVT in summer months. However, larger studies are needed to confirm definite associations.

In-vitro and in-vivo anticandidal activity of Trachyspermum ammi (L.) sprague seeds ethanolic extract and thymol-containing hexanes fraction.

Candida albicans is the causative agent of candidiasis in immunocompromised and diseased individuals. The current study was designed to evaluate the antifungal activity of Trachyspermum ammi (L.) Sprague seeds ethanolic extract and hexanes fraction against Candida albicans, both in-vitro and in-vivo. The minimal inhibitory concentration of hexanes fraction was found to be 225 µg/mL, as compared to the standard drug amphotericin B (200 µg/mL). For the treatment of cutaneous candidiasis in BALB/c mice model, the extract and fraction containing ointment was topically applied on flank of mice at various concentrations, and 90-100% recovery in mice was observed which was better than the standard drug, clotrimazole. The GC/MS analysis of hexanes extract indicated the presence of three major components, including (43.91%), O-cymene (25.53%), and γ-terpinene (22.64%). The results showed that T. ammi possess potential antifungal effect, and can be used for the development of novel antifungal agents after further pre-clinical and clinical trials.

Mesomeric Effects of Azobenzene Bearing Natural Product-Based Molecules for Liquid Crystal Materials: An Overview.

Latest progress in the liquid crystal (LC) field related to azo molecules incorporated into natural product- based moieties for the improvement of LC texture and mesomeric phases has received great interest among researchers. A LC containing natural product-based moieties i.e. menthol, kojic acid, cholesterol and chalcone with stable azo and azobenzene scaffolds with specific optical tunability, has been widely used in photo-active materials such as Liquid Crystal Display (LCD), LC films, smart windows and other devices. This review discusses the influence of azobenzene, a renowned photo-responsive and stable LC scaffold, in mesogenic phases due to photo-isomerization and optical switching. The incorporation of mesomeric phases of natural product moieties to azo molecules has improved the properties of LC, i.e, from the nematic phase to the smectic phase with proper magnetic field alignment. Natural product-based LC can be useful in numerous applications, especially practical electronic or optic devices such as optical image storage, display devices, solar cells, optical switching.

Characterization of bacterial community structure in the rhizosphere of Triticum aestivum L.

The plant microbiome influence plant health, yield and vigor and has attained a considerable attention in the present era. In the current study, native bacterial community composition and diversity colonizing Triticum aestivum L. rhizosphere at two distant geographical locations including Mirpur Azad Kashmir and Islamabad was elucidated. Based on IonS5™XL platform sequencing of respective samples targeting 16S rRNA gene that harbor V3-V4 conserved region revealed 1364 and 1254 microbial operational taxonomic units (OTUs) at ≥97% similarity and were classified into 23, 20 phyla; 70, 65 classes; 101, 87 orders; 189,180 families; 275, 271 genera and 94, 95 species. Respective predominant phyla accounting for 97.90% and 98.60% of bacterial community were Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Firmicutes, Chloroflexi and Gemmatimonadetes. Diversity indices revealed variations in relative abundance of bacterial taxa owing to distant geographical locations however predominant bacterial taxa at both locations were similar. These findings paved a way to dissect consequence of associated microbiota on future wheat production system.

Antimicrobial and synergistic activity of thiazoline derivatives in combination with conventional antibiotics against multidrug resistant Staphylococcus aureus isolated from abscess drainage samples.

Emergence and spread of multidrug resistant (MDR) Staphylococcus aureus strains is becoming major challenge in treatment of soft tissue infections. This study aimed to explore antimicrobial and synergistic antimicrobial potential of three commercially available thiazoline derivatives (2-amino-2-thiazoline, 2-thiazoline-2-thiol and 2-acetyl-2-thiazoline) against MDR Staphylococcus aureus strains isolated from abscess drainage samples (n=20). MDR Staphylococcus aureus isolates were identified by Kirby-Bauer disk diffusion assay and were further subjected to molecular identification by 16srRNA amplification and DNA sequencing. Minimum Inhibitory Concentration (MIC) values of test compounds and antibiotics (0.25-512µg/mL) were measured and subsequently, synergism assay was performed to calculate Fractional Inhibitory Concentration (FIC) index. Out of twenty Staphylococcus aureus isolates, sixteen (80%) were found to be MDR whereas four (20%) were Non-MDR. Moxifloxacin and vancomycine were found most effective antibiotics, inhibiting 100% (n=20) and 95% (n=19) strains respectively. Antimicrobial activity of 2-amino-2-thiazoline (MIC: 32µg/mL), 2-thiazoline-2-thiol (MIC: 64µg/mL) and 2-acetyl-2-thiazoline (MIC: 32µg/mL) was found significant against all ten tested MDR strains. Synergistic combinations of thiazoline derivatives with test antibiotics reduced MIC values significantly. Therefore, combination of tested thiazoline derivatives with antibiotics could be used as alternative therapeutic approach to treat soft tissue infections caused by MDR Staphylococcus aureus after further pre-clinical and clinical studies.

Probiotic potential of Saccharomyces strains isolated from Litchi chinensis (Lychee fruit).

Recently, probiotic yeasts have become an interesting topic of research all over the world. Saccharomyces cerevisiae is well proven probiotic yeast against several gastrointestinal diseases. Current study aimed to explore the probiotic potential and antibacterial properties of Saccharomyces strains isolated from fresh lychee fruits available in local markets of Karachi, Pakistan. Probiotic potential and antibacterial activity of locally isolated probiotic yeast strains (named as S. cerevisiae BEL 1 and S. cerevisiae BEL 9) was studied against gastrointestinal pathogens using standard in vitro screening methods. Comparative analysis was also carried out with commercially available S. boulardii probiotic preparations. Furthermore, for probiotic potential, all the studied yeast strains were exposed to various stress conditions inherent of gastrointestinal tract i.e., thermo tolerance, pH tolerance, bile salts survivability and osmo-tolerance. Isolated strains (BEL 1 and BEL 9) were able to tolerate at the temperatures (40oC and 45oC), moreover survived in the presence of gastric juices, extreme bile salt concentrations (range 0.5%-2%) and different osmotic stress conditions (1M and 1.5 M NaCl). Optimal growth was observed at 37oC. Similar growth pattern and viability of BEL 1 and 9 was found for most of the stress conditions, when compared with the commercially available strains of S. boulardii. Therefore, isolated yeast strains BEL 1 and 9 will be considered as a potential bio-therapeutic agent because of the promising probiotic potential.

Case series of rare Interstitial Lung Disease (ILD).

Pulmonary alveolar proteinosis (PAP) is a rare lung disease where periodic acid Schiff (PAS)-positive eosinophilic material accumulates in the alveoli of the lungs. Here we describe two cases of young males who presented with dynpnoea and weight loss. The HRCT scan of the chest in both cases showed the typical "crazy-paving" pattern and lung biopsies confirmed the diagnosis of PAP. They showed remarkable symptomatic improvement with therapeutic whole lung lavage.

Harmaline and its Derivatives Against the Infectious Multi-Drug Resistant Escherichia coli.

BACKGROUND: Multidrug resistance (MDR) is a major challenge in the treatment of infectious diseases. The MDR in urinary tract infection causing bacteria, such as Escherichia coli, has made treatment of UTI very difficult. OBJECTIVE: The aims of the current study were to synthesize a library of harmaline derivatives, and to evaluate their activity against various strains of multi-drug resistance (MDR) E. coli. METHOD: Harmaline derivatives were synthesized by the reaction of harmaline (1) with various acid halides and anhydrides. These compounds were subjected to susceptibility determination by in vitro MTT assay. The changes in morphology of the bacterial cells after the treatment with harmaline (1) and its new derivatives 2 and 3 were studied through scanning electron, atomic force and fluorescence microscopy. Effect of harmaline and its derivatives on the production of Reactive Oxygen Species (ROS) in MDR E. coli was assessed through lucigenin chemiluminescence assays. RESULTS: The selected compounds assisted the fluorescently labeled dye DiBAC4(3) to bind to the lipid rich intra-cellular entities, and thus produced a sharp green fluorescence by easily penetrating into the compound-induced depolarized membrane of MDR E. coli. These compounds have also triggered a significant generation of ROS from bacterial cells as compared to the conventional antibiotics. The current study demonstrated that harmaline (1), and its derivatives 2 and 3 were identified as anti-MDR agents against MDR strains of E. coli. Antibacterial effect of compounds 1-3 on MDR E. coli is possibly due to membrane depolarization due to ROS-induced damage to the bacterial cell membrane. CONCLUSION: Harmaline and its derivatives were identified as anti-MDR agents against various highly resistant and Pakistani MDR clinical isolates of E. coli. These compounds may serve as the leads for further studies towards the development of treatment against the infections caused by MDR E. coli.

Self-medication with antibiotics among non-medical university students of Karachi: a cross-sectional study.

BACKGROUND: The prevalence of self -medication with antibiotics is quite high in developing countries as opposed to developed countries. Antibiotics are often taken erroneously for certain ailments, without having the appropriate knowledge of their use. This carries potential risks for the individual as well as the community, in form of several side effects such as antibiotic resistance. Therefore the prevalence of self-medicated antibiotics in developing countries needs to be studied. METHODS: A descriptive cross-sectional study was carried out at six different non-medical universities of Karachi. 431 students were included in the study. Data was collected using self-administered questionnaires and analyzed using SPSS version 19. RESULTS: 50.1% students reported having self-medicated themselves in the past 6 months and 205 (47.6%) reported self-medication with antibiotics. Amoxicillin was the most self-prescribed antibiotic (41.4%). Awareness of the adverse effects of antibiotics was demonstrated by 77.3% of the students and sleep disturbance was the most commonly known (46.5%) side effect. 63.1% denied having any knowledge about antibiotic resistance and only 19.9% correctly knew that indiscriminate use of antibiotics can lead to increased antibiotic resistance. CONCLUSION: The prevalence of self-medication with antibiotics among the non-medical university students was high despite the awareness of adverse effects. Antibiotic resistance was a relatively unknown terminology.