Abstract: Objective: To investigate the role of CRX-527, a Toll-like receptor 4 agonist, as the possible adjuvant for recombinant Mycobacterium bovis Bacillus Calmette-Guerin expressing merozoite surface protein 1C (BCG-MSP-1C). Methods: The mice were immunized with BCG and BCG-MSP- 1C in the presence and absence of CRX-527. The untreated mice (injected with PBS-T80 only) were the negative control. The ability of CRX-527 to enhance IgG and its subclasses, as well as IL-4 and IFN-γ production in the serum and spleen supernatant was evaluated using ELISA. Results: Mice immunized with BCG-MSP-1C exhibited the highest production of IgGs, IL-4 and IFN-γ after third immunization. In addition, CRX-527 further promoted the production of total IgG and IgG subclasses as well as IFN-γ and IL-4 in the serum and splenocytes of immunized mice. Conclusions: CRX-527 has the potential as an adjuvant candidate for the candidate vaccines. Further study is needed to verify appropriate dosage for immunization and its efficacy.

Purpose: We investigated research barriers among Jordanian medical postgraduates to understand the current context of the local health research landscape and improve scholarly output. Methods: Using a validated questionnaire, Jordanian interns, residents, specialists, and consultants were examined for their perceived attitudes and barriers towards research. Participants were conveniently sampled from public, university, military, and private institutions. Differences in responses were examined using the Student t-test and analysis of variance. Binary logistic regression was utilized to examine predictors of being able to publish. Results: A total of 1,141 Jordanian medical postgraduates were recruited, of which 61.3% were junior postgraduates (i.e., interns and residents in their first 2 years of residency) while 38.7% were senior postgraduates (i.e., senior residents, specialists, and consultants). Around 76.0% of participants had no peer-reviewed publications. Of those with least one publication (n=273), only 31.1% had first authorships. Participants portrayed dominantly positive attitudes towards the importance of research. There were no significant differences between junior and senior postgraduates for overall attitudes (p=0.486) and knowledge barriers scores (p=0.0261). Conversely, senior postgraduates demonstrated higher mean organizational barriers (p<0.001). Seniority (odds ratio [OR], 5.268; 95% confidence interval [CI], 3.341–8.307), age (OR, 1.087; 95% CI, 1.019–1.159), academic standing (OR, 1.730; 95% CI, 1.103–2.715), and confidence (OR, 1.086; 95% CI, 1.009–1.169) were positive predictors of publication in peer reviewed journals. Conclusion The Jordanian medical research landscape is riddled with all forms of different barriers. The reworking of current and integration of new research training programs are of utmost importance.

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia.
Asthenozoospermia/pathology* ; Dyneins/genetics* ; Homozygote ; Humans ; Male ; Microtubule-Associated Proteins ; Mutation ; Mutation, Missense ; Sperm Tail/metabolism*

Aims: The study was aimed to isolate and characterize the mycotoxin-producing filamentous Aspergillus parasiticus from the feed samples. The sensitivity pattern of the isolates was assessed against different disinfectants. Methodology and results: Fifty different feed samples were screened for A. parasiticus isolation. Isolates were subjected to macroscopic and microscopic characterization. Polymerase chain reaction was performed to confirm the isolates at the genomic level. Mycotoxin producing potential of the isolates was assessed by thin-layer chromatography (TLC). To quantify the toxins, high performance liquid (HPLC) was employed. The antifungal potential of disinfectants was determined by the well diffusion method followed by minimum inhibitory concentration (MIC) calculation. Out of twenty isolates of A. parasiticus, 11(55%) isolates were observed positive for toxin production. Three toxigenic isolates (AspP2, AspP4 and AspP8) were selected to evaluate their susceptibility against disinfectants by well diffusion method. AspP2 produced maximum (5.90 ng/mL) toxin, followed by AspP4 (3.11 ng/mL) and AspP8 (18.47 ng/mL). Terralin showed maximum fungicidal activity with 29.66 ± 8.08 mm zone of inhibition at 0.42 μg/mL MIC. Hypochlorite and Instru Star showed 99% disinfection with 30, 60 and 90 min contact time (6 mean log reduction) for all A. parasiticus isolates. Alpha Guard inhibited growth after 15 min contact time for all the isolates. Conclusion, significance and impact of study This study provides data indicating the contamination of feed samples with mycotoxin-producing A. parasiticus isolates and their sensitivity against commercially available disinfectants. Use of these disinfectants in appropriate concentrations and time could help prevent the contamination of food, feed and healthcare settings with the fungal species.
Mycotoxins ; Aspergillus

The study aimed to achieve enhanced targeted cytotoxicity and cell-internalization of cisplatin-loaded deoxyribonucleic acid-nanothread (CPT-DNA-NT),mediated by scavenger receptors into HeLa cells.DNA-NT was developed with stiff-topology utilizing circular-scaffold to encapsulate CPT.Atomic force microscopy (AFM) characterization of the DNA-NT showed uniformity in the structure with a diameter of 50-150 nm and length of 300-600 nm.The successful fabrication of the DNA-NT was confirmed through native-polyacrylamide gel electrophoresis analysis,as large the molecular-weight (polymeric) DNA-NT did not split into constituting strands under applied current and voltage.The results of cell viability confirmed that blank DNA-NT had the least cytotoxicity at the highest concentration (512 nM) with a viability of 92％ as evidence of its biocompatibility for drug delivery.MTT assay showed superior cyto-toxicity of CPT-DNA-NT than that of the free CPT due to the depot release of CPT after DNA-NT inter-nalization.The DNA-NT exhibited targeted cell internalizations with the controlled intracellular release of CPT (from DNA-NT),as illustrated in confocal images.Therefore,in vitro cytotoxicity assessment through flow cytometry showed enhanced apoptosis (72.7％) with CPT-DNA-NT (compared to free CPT;64.4％).CPT-DNA-NT,being poly-anionic,showed enhanced endocytosis via scavenger receptors.

Objective: To determine the effects of toll-like receptor 2 (TLR-2) agonist, Pam3CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin (rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum. Methods: Six groups of mice (n=6 per group) received intraperitoneal phosphate buffered saline T80 (PBS-T80), BCG or rBCG in the presence or absence of Pam3CSK4. Enzyme-linked immunosorbent assay was carried out to measure serum total IgG, IgG1, IgG2a, and IgG2b production. Spleens were also harvested and splenocytes were co-cultured with rBCG antigen for in vitro determination of IL-4 and IFN-γ via enzyme-linked immunosorbent assay. Results: The production of total IgG and the isotype IgG1, IgG2a and IgG2b was significantly higher in rBCG-immunised mice than in the BCG and PBS-T80-immunised mice, and Pam3CSK4 further enhanced their productions. A similar pattern was also observed in IFN-γ production. Moreover, there was no significant difference in IL-4 production in all groups either in the presence or absence of Pam3CSK4. Conclusions: We present evidence of the adjuvant effects of TLR-2 agonist in enhancing the production of total IgG, IgG1, IgG2a, IgG2b, as well as IFN-γ in response to rBCG. However, the presence or absence of Pam3CSK4 had no effect on IL-4 production.

Objective: To determine the role of toll-like receptor 4 (TLR-4) in eliciting cellular and humoral immune responses against recombinant Mycobacterium bovis bacille Calmette-Guérin (rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum. Methods: Six groups of mice (n=6 per group) were injected with phosphate buffered saline T80, BCG or rBCG intraperitoneally, in the presence or absence of a TLR-4 inhibitor; TAK-242. Enzyme-linked immunosorbent assay was carried out for serum total IgG, IgG1, IgG2a and IgG2b determination. Spleens were also harvested and splenocytes cultured for determination of intracellular cytokines; IL-4 and IFN-γ via enzyme-linked immunosorbent assay. Results: The production of total IgG, and the subclasses IgG1, IgG2a and IgG2b was significantly higher in rBCG-immunised mice than BCG and phosphate buffered saline immunised mice in the absence of TAK-242. A significant rise in total IgG occurred with more booster immunisations. The level of IgG2a was highest, followed by IgG2b, then IgG1. The production of both IL-4 and IFN-γ was also highest in the rBCG immunised groups. These significant rises were inhibited in the presence of TAK-242. Conclusions: We present evidence of the role of TLR-4 in the increased production of total IgG, IgG1, IgG2a and IgG2b, as well as IL-4 and IFN-γ in response to our rBCG construct.

Scabies occurs in human due to the burrowing ectoparasite Sarcoptes scabiei var. hominis resulting in intense itching and inflammation, and manifesting as a skin allergy. Limited information is available about the genetic diversity of S. scabiei in human. In this study, we characterized S. scabiei var. hominis using nuclear marker ITS-2, mitochondrial marker 16S and microsatellite markers. To examine the extent of the genetic variation, individual mite gDNA was first amplified using ITS-2, 16S and microsatellite primers by PCR, later amplicons were sequenced directly and analysed by MEGA 7. Sequence analysis of ITS- 2 showed no host segregation and geographical isolation, whereas 16S indicated presence of both hosts adapted and geographically segregated populations of S. scabiei. Results of microsatellites revealed polymorphism as allelic variability between and within the populations studied. The different varieties of Sarcoptes mites belong to different host species and geographic regions recommended that Sarcoptes mites are genetically isolated. This is the first report on the molecular characterization of S. scabiei var. hominis from Pakistan. Additionally, genetic studies including a greater number of specimens are required to comprehend the molecular epidemiology of Sarcoptes mite in Pakistan.

Aims: To study the performance of SMFC in the terms of power generation and toxic metals removal. This study was also focused on the characterization of SMFC electro-microbiology. Methodology and results: A SMFC was designed and loaded with sediment and overlying water. This SMFC was synchronized with wireless data logger acquisition system. The toxic metals removal capacity was measured by atomic absorption spectroscopy. The characterization of SMFC bacteria was done by 16S rRNA.In this study the experiments were carried out in a dual-chamber SMFC with external resistances 30 kΩ-50 Ω. The SMFC was produced power about 630 mV with maximum power density 40 mW/m2and current density 250 mA/m2. After 120 days of operation, SMFC removed cadmium and copper about 22.6 and 150 mg/kg, respectively. The SMFC also showed high cadmium (86%) and copper (90%) removal at pH 7.0 and temperature 40 °C. The most dominant bacterial community at anode and cathode was identified as Pseudomonas spp. which could be function as exoelectrogen. Conclusion, significance and impact of the study: The results indicated that the SMFC system could be applied as a long term and effective tool for the removal of cadmium and copper contaminated sediments and supply power for commercial devices. The Pseudomonas spp. may be used as a genetic donor for the other non-exoelectrogens strains.