Good Quality Locally Procured Drugs Can Be as Effective as Internationally Quality Assured Drugs in Treating Multi-Drug Resistant Tuberculosis

Background

Owing toGiven the high costs of drugs to treat multi-drug resistant tuberculosis (MDR-TB), the Green Light Committee (GLC) initiative enables TB programs to procure quality-assured drugs at reduced prices. Despite price reductions, internationally quality assured (IQA) drugs can be more expensive than locally procured drugs. There is little evidence to inform decision-makers about whether IQA drugs are more effective than local drugs. This is the first study to compare outcomes between MDR-TB patients treated using IQA, and locally procured drugs in the same hospitals during the same time period.

Methods/Findings

A retrospective cohort study was conducted in three hospitals across Pakistan. Data on baseline characteristics and treatment outcomes during the first six months of treatment were extracted from hospital records of adult culture-positive pulmonary MDR-TB patients starting treatment between January 2011 and June 2012. Two cohorts were defined: patients receiving IQA drugs, and patients receiving locally procured non-IQA drugs. Data were analysed using Kaplan-Meier curves and Cox proportional hazards regression. The primary outcome compared between cohorts was time to culture conversion. Of 231 patients, 90 were in the IQA and 141 in the non-IQA cohorts. Baseline characteristics were similar except for higher frequency of quinolone resistance in the IQA cohort. Overall, 193 patients (84%) culture converted. Culture conversion was not faster in the IQA cohort; the median time was 81 and 68 days in the IQA and non-IQA cohorts, respectively. Unadjusted and adjusted hazard ratios for culture conversion in IQA verses non-IQA cohorts were 0.82 (95%-CI, 0.62-1.10) and 0.95 (95%-CI, 0.66-1.36) respectively.

Conclusions

Use of good quality, locally procured drugs can be effective in treating MDR-TB, may involve lower costs than using IQA drugs and could strengthen developing country drug quality assurance systems. This may be a suitable alternative in lieu of or whilst awaiting arrival of internationally procured medicines.     

Antioxidant Potential and In Situ Analysis of Major and Trace Element Determination of Ood-saleeb,a Known Unani Herbal Medicine by ICP-MS

The intention of the present research work was to investigate the antioxidant activity and trace element analysis of Ood-saleeb, a known herbal medicine. Preliminary screening of phytochemicals showed that the extract of Ood-saleeb had flavonoids and phenolics. The significant activities in all antioxidant assays were observed in the extract of Ood-saleeb in comparison with the standard antioxidant with respect to dose of Ood-saleeb. Incredible activities to scavenge reactive oxygen species were also observed by the extract of Ood-saleeb. The IC₅₀ values of all factors were determined using ascorbic acid as a standard. The inductive coupled plasma-mass spectroscopy (ICP-MS) was employed for the estimation of trace elements in Ood-saleeb extract. The concentrations of up to 18 elements were detected successfully. Silicon was found in high concentration (85.3 μg/g) while lithium was in low concentration (3 ng/g). The trace elements in the sample were found at different percentage levels which play a key role in the treatment of diseases.     

Mycorrhiza and Phosphate Solubilizing Bacteria: Potential Bioagents for Sustainable Phosphorus Management in Agriculture

Phosphorus (P) is a critical nutrient that plays an essential role in improving soil fertility for optimum plant growth and productivity. It is one of the most deficient macro-nutrients in agricultural soils after nitrogen and is considered inadequate for plant growth and production. To P availability in soils, the farmers are applying huge amounts of synthetic P fertilizers that adversely affect the wider environment, groundwater, soil fertility and microbial population. Many beneficial microbes are known to release and supply soluble P for improving growth and yield of a variety of plants in a sustainable manner in P deficient soils. Thus, inoculation of these microbes, including arbuscular mycorrhizal fungi (AMF) and phosphate solubilizing bacteria (PSB) to soil to enhance crop production without harming the environment, is an alternative approach to chemical fertilizers. The combined role of AMF and PSB in P solubilization is not well understood and the application and mode of action of these microbial groups are often naive due to variation in the environment. Therefore, the current review article would develop a better understanding of the interactive role and mechanisms of AMF and PSB in improving P availability from both organic and inorganic sources in a sustainable crop production system. Finally, the current review would loop out further avenues for researchers interested to commercially produce effective AMF and PSB-based biofertilizers for sustainable management of phosphorus over a wide range of agricultural crops worldwide.     

Kaolin and Jasmonic acid improved cotton productivity under water stress conditions

Drought is one of the most emerging threat that causes a severe reduction in cotton plant growth and development. Being cotton is a major cash crop has great threat to prevailing drought events in Pakistan. A field experiment was conducted in Kharif season 2018 at Research Area of MNS-University of Agriculture, Multan, Pakistan to assess the role of foliar applied kaolin and jasmonic acid on vegetative growth, gas exchange and reproductive traits of cotton under normal irrigated and artificial water deficit conditions. The experiment was laid -out in a factorial randomized complete block design with split – split plot arrangement. Main plots were allocated for irrigation levels, sub-plots for two -cotton genotypes viz. NIAB − 878 and SLH − 19 while sub – sub plots for treatments of kaolin and Jasmonic acid. Water deficit stress was created by skipping irrigation at flowering for 21 days. Foliar sprays of Kaolin (5%, w/v) and Jasmonic acid (100 μM) were applied alone or in combination at 60 days after planntinon both to normal irrigated and water-stresse skip irrigation while irrigation water alone was sprayed in control plots. Both cotton genotypes responded variably to normal irrigated and skip conditions. Skipping irrigation for up to 21 days at flowering caused a significant decrease in leaf relative water content, SPAD values, net photosynthetic rate and seed cotton yield in both the genotypes. Seed cotton yield showed an overall decline of 24.7% in skip over Normal irrigated crop. The genotype NIAB − 878 produced maximum seed cotton yield of 3.304 Mg ha⁻¹ in normal that dropped to 2.579 Mg ha⁻¹ in skip, thus showing an average decline of 21.9 %. Similarly, SLH − 19 produced 2.537 Mg ha⁻¹ seed cotton under normal that dropped to 1.822 Mg ha⁻¹ in skip, showing an average decline of 28.2%. The Application of Kaolin and JA Jasmonic acid, either applied individually or in combination, improved vegetative and reproductive development of both cotton varieties in normal and skip regimes. However, combined kaolin and Jasmonic Acid application proved to be more beneficial in terms of seed cotton production and other parameters studied.     

Ethnomedicinal assessment of Irula tribes of Walayar valley of Southern Western Ghats,India

The present study was aimed to explore the traditional knowledge of Irula tribal people who are practicing herbal medicine in Walayar valley, the Southern Western Ghats, India. A total number of 146 species of plants distributed in 122 genera belonging to 58 families were identified as commonly used ethnomedicinal plants by them. Interestingly, 26 new claims were also made in the present study. Through the data obtained from Irula tribal healers, the herbs were mostly used for medicine (40.4%) followed by trees (26.7%) and climbers (18.5%). In addition leaves were highly used for medicinal purposes, collected from 55 species (38%) followed by multiple parts from 18 species (12%). Acorus calamus is the species of higher use value (1.80) assessed to be prescribed most commonly for the treatment of cough. High informant consensus factor (1.0) obtained for insecticidal uses and cooling agent indicates that the usage of Canarium strictum and Melia dubia, and Mimosa pudica and Sesamum indicum respectively for that purposes had obtained high degree of agreement among the healers in using these species for the respective purposes. The most commonly used method of preparation was decoction (63%) followed by raw form (23%), paste (12%) and powder (2%). Therefore, it is suggested to take-up pharmacological and phytochemical studies to evaluate the species to confirm the traditional knowledge of Irulas on medicinal plants.     

A comparative gametogenesis and biochemical differentiation of goatfish (Family: Mullidae) from the Red Sea

Comparative study between few species of goatfishes (Mullidae), locally called Sultan Ibrahim, has been investigated from Jeddah coast. The present study compares gonad structure, oogenesis and spermatogenesis in three species of goatfish. The various stages are similar in the different species, but differ in time of spawning. The reproductive biology of three goatfishes, Mulloidichthys flavolineatus (1 black blotch), Mulloidichthys flavolineatus (3 black blotches), Mulloidichthys vanicolensis, were examined. A macroscopic and histological examination of the gonads demonstrated that all the three species are typically gonochoristic. Species-specific reproductive parameters including male stages, changes in gonado-somatic index (GSI), classification of oocyte stages were analyzed for each of the three targeted species. All three species have asynchronous ovary. The spawning season for the three studied species differ from one species to another i.e., the spawning season for M. vanicolensis females and males from April to July; M. flavolineatus, (1 black blotches) for both sexes from May until December; for M. flavolineatus (3 black blotches) showed a peak in May to early October for both sexes. Ultrastructurally the mature spermatozoon’s head shape appears like a computermouse, the tiny mid-piece consist two mitochondria and a flagellum. The chromatin, within the nucleus is electron-dense and homogenous in M. flavolineatus (with one black blotch), while the chromatin is granular in (M. vanicolensis and M. flavolineatus (3 black blotches)). The posterior region of the nucleus is deeply penetrated by the nuclear fossa which contains, the basal part of flagellum and centriolar complex. The ultrastructure of the eggs of the three Mullidae species is circular in shape and contains a simple, circular micropyle. The ovarian follicular epithelium of the three Mullidae species consists of two layers, an outer theca layer and an inner granulosa layer with zona radiata under them. Zona radiata consists of 2 layer-interna and externa. Zona radiata interna is thicker than zona radiata externa. Differences in protein patterns of muscles and gonads were observed in the present study.     

Exosomes Derived from HIV-1-infected Cells Contain Trans-activation Response Element RNA

Exosomes are nano-sized vesicles produced by healthy and virus-infected cells. Exosomes derived from infected cells have been shown to contain viral microRNAs (miRNAs). HIV-1 encodes its own miRNAs that regulate viral and host gene expression. The most abundant HIV-1-derived miRNA, first reported by us and later by others using deep sequencing, is the trans-activation response element (TAR) miRNA. In this study, we demonstrate the presence of TAR RNA in exosomes from cell culture supernatants of HIV-1-infected cells and patient sera. TAR miRNA was not in Ago2 complexes outside the exosomes but enclosed within the exosomes. We detected the host miRNA machinery proteins Dicer and Drosha in exosomes from infected cells. We report that transport of TAR RNA from the nucleus into exosomes is a CRM1 (chromosome region maintenance 1)-dependent active process. Prior exposure of naive cells to exosomes from infected cells increased susceptibility of the recipient cells to HIV-1 infection. Exosomal TAR RNA down-regulated apoptosis by lowering Bim and Cdk9 proteins in recipient cells. We found 10⁴–10⁶ copies/ml TAR RNA in exosomes derived from infected culture supernatants and 10³ copies/ml TAR RNA in the serum exosomes of highly active antiretroviral therapy-treated patients or long term nonprogressors. Taken together, our experiments demonstrated that HIV-1-infected cells produced exosomes that are uniquely characterized by their proteomic and RNA profiles that may contribute to disease pathology in AIDS.     

Induction of apoptosis by pyrazolo[3,4-d]pyridazine derivative in lung cancer cells via disruption of Bcl-2/Bax expression balance

In the rapidly expanding era of cancer target therapy, regulators of apoptosis are emerging as attractive therapeutic targets. X-linked inhibitor of apoptosis (XIAP) is of specific interest owing to its characteristic overexpression in a wide variety of neoplasms, with a resultant survival advantage for tumor cells and treatment resistance. In this study, we examined three pyrazolo [3,4-d] pyridazine derivatives (PPDs) through molecular modeling and studied their modes of interaction with XIAP-BIR3 domain. PPD-1, which possessed the highest binding affinity with XIAP, was tested on A549 (lung cancer cell line); HCT-116 (colorectal carcinoma cell line); HEPG2 (liver carcinoma cell line), HFB4 (normal human skin melanocyte cell line) and WI-38 (human embryonic lung fibroblasts). In comparison to cisplatin as a positive control, PPD-1 yielded remarkable cytotoxicity on all cancer cell lines, with the highest anti-tumor activity on A549 and a favorable therapeutic ratio. Flow cytometry studies concluded that PPD-1 treatment induces Sub G1 and G2/M cell cycle arrest and apoptosis. The percentage of apoptotic cells in PPD-1 treated A549 cells was considerably higher than that in untreated cells (10.06% vs 0.57%, respectively). To further investigate the mechanism of induction of apoptosis by PPD-1, Real time-PCR was used to quantify the expression levels of key apoptotic regulators. Significant overexpression of the effector capsase-3, pro-apoptotic bax and tumor suppressor gene p53 were noted as compared to untreated cells (7.19 folds, 7.28 folds, and 5.08 folds, respectively). Moreover, PPD-1 inhibited the expression of the anti-apoptotic bcl-2 gene to 0.22 folds. These findings demonstrate that PPD-1 treatment disrupts the Bcl-2/BAX balance in lung cancer cell lines, leading to apoptosis induction possibly through intrinsic mitochondria-dependent pathway. These novel insights elucidate the mechanism of PPD-1 cytotoxicity in lung cancer cell lines and offer a promising therapeutic approach that needs further study.     

Effervescence Assisted Fusion Technique to Enhance the Solubility of Drugs

The solubility of five poorly soluble drugs was enhanced by using an effervescence assisted solid dispersion (EASD) technique. EASDs were prepared by using modified fusion method. Drug and hydrophilic carrier were melted, and in this molten mixture, effervescence was generated by adding effervescence couple comprising organic acid (citric acid) and carbonic base (sodium bicarbonate). Solubility of drug powders, solid dispersions, and EASDs was determined at 25°C using shake flask method. Atorvastatin calcium, cefuroxime axetil, clotrimazole, ketoconazole, and metronidazole benzoate were estimated using a spectrophotometer at 246, 280, 260, 230, and 232 nm (λ_max), respectively. Solubility of atorvastatin calcium (from 100 to 345 μg/ml), cefuroxime axetil (from 441 to 1948 μg/ml), clotrimazole (from 63 to 677 μg/ml), ketoconazole (from 16 to 500 μg/ml), and metronidazole benzoate (from 112 to 208 μg/ml) in EASDs was enhanced by 3.45-, 4.4-, 10.7-, 31.2-, and 1.8-fold, respectively. Scanning electron micrographs of drug powder, solid dispersion, and EASDs were compared. Scanning electron micrographs of EASDs showed a uniform distribution of drug particles in the carrier matrix. Morphology (size and shape) of cefuroxime axetil particles was altered in solid dispersion as well as in EASD. EASDs showed better solubility enhancement than conventional solid dispersions. The present technique is better suitable for drugs having a low melting point or melt without charring. Effervescence assisted fusion technique of preparing solid dispersions can be employed for enhancing solubility, dissolution, and bioavailability of poorly soluble drugs.KEY WORDS: dissolution, effervescence, fusion, solid dispersion, solubility     

Genetic diversity and prevalence of CCR2-CCR5 gene polymorphisms in the Omani population

Polymorphisms in the regulatory region of the CCR5 gene affect protein expression and modulate the progress of HIV-1 disease. Because of this prominent role, variations in this gene have been under differential pressure and their frequencies vary among human populations. The CCR2V64I mutation is tightly linked to certain polymorphisms in the CCR5 gene. The current Omani population is genetically diverse, a reflection of their history as traders who ruled extensive regions around the Indian Ocean. In this study, we examined the CCR2-CCR5 haplotypes in Omanis and compared the patterns of genetic diversity with those of other populations. Blood samples were collected from 115 Omani adults and genomic DNA was screened to identify the polymorphic sites in the CCR5 gene and the CCR2V64I mutation. Four minor alleles were common: CCR5-2554T and CCR5-2086G showed frequencies of 49% and 46%, respectively, whereas CCR5-2459A and CCR5-2135C both had a frequency of 36%. These alleles showed moderate levels of heterozygosity, indicating that they were under balancing selection. However, the well-known allele CCR5Δ32 was relatively rare. Eleven haplotypes were identified, four of which were common: HHC (46%), HHE (20%), HHA (14%) and HHF*2 (12%).