Phyto-Synthesis, Characterization, and In Vitro Antibacterial Activity of Silver Nanoparticles Using Various Plant Extracts.

Aloe vera, Mentha arvensis (mint), Coriandrum sativum (coriander), and Cymbopogon citratus (lemongrass) leaf extracts were used to synthesize stable silver nanoparticles (Ag-NPs) by green chemistry. UV-vis spectrophotometry, X-ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) spectroscopy techniques were used to characterize these biosynthesized nanoparticles. The data indicated that the silver nanoparticles were successfully synthesized, and the narrower particle size distribution was at 10-22 nm by maintaining a specific pH. As a short-term post-sowing treatment, Ag-NP solutions of different sizes (10 and 50 ppm) were introduced to mung bean seedlings, and the overall increase in plant growth was found to be more pronounced at 50 ppm concentration. The antibacterial activity of Ag-NPs was also investigated by disc diffusion test, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) test. The zones of inhibition (ZOI) were shown by Escherichia coli (E. coli) (1.9, 2.1, 1.7, and 2 mm), followed by Staphylococcus aureus (S. aureus) (1.8, 1.7, 1.6, and 1.9 mm), against coriander, mint, Aloe vera, and lemongrass, respectively. MIC and MBC values of E. coli, and S. aureus ranged from 7 to 8 µg/mL. Overall, this study demonstrates that Ag-NPs exhibit a strong antimicrobial activity and thus might be developed as a new type of antimicrobial agent for the treatment of bacterial infection.

CDC14A phosphatase is essential for hearing and male fertility in mouse and human.

The Cell Division-Cycle-14 gene encodes a dual-specificity phosphatase necessary in yeast for exit from mitosis. Numerous disparate roles of vertebrate Cell Division-Cycle-14 (CDC14A) have been proposed largely based on studies of cultured cancer cells in vitro. The in vivo functions of vertebrate CDC14A are largely unknown. We generated and analyzed mutations of zebrafish and mouse CDC14A, developed a computational structural model of human CDC14A protein and report four novel truncating and three missense alleles of CDC14A in human families segregating progressive, moderate-to-profound deafness. In five of these families segregating pathogenic variants of CDC14A, deaf males are infertile, while deaf females are fertile. Several recessive mutations of mouse Cdc14a, including a CRISPR/Cas9-edited phosphatase-dead p.C278S substitution, result in substantial perinatal lethality, but survivors recapitulate the human phenotype of deafness and male infertility. CDC14A protein localizes to inner ear hair cell kinocilia, basal bodies and sound-transducing stereocilia. Auditory hair cells of postnatal Cdc14a mutants develop normally, but subsequently degenerate causing deafness. Kinocilia of germ-line mutants of mouse and zebrafish have normal lengths, which does not recapitulate the published cdc14aa knockdown morphant phenotype of short kinocilia. In mutant male mice, degeneration of seminiferous tubules and spermiation defects result in low sperm count, and abnormal sperm motility and morphology. These findings for the first time define a new monogenic syndrome of deafness and male infertility revealing an absolute requirement in vivo of vertebrate CDC14A phosphatase activity for hearing and male fertility.

Epidemiology, major risk factors and genetic predisposition for breast cancer in the Pakistani population.

Occurrence of breast cancer is related to genetic as well as cultural, environmental and life-style factors. Variations in diversity of these factors among different ethnic groups and geographical areas emphasize the immense need for studies in all racial-ethnic populations. The incidence of breast cancer in Pakistan is highest in Asians after Jews in Israel and 2.5 times higher than that in neighboring countries like Iran and India, accounting for 34.6% of female cancers. The Pakistani population is deficient in information regarding breast cancer etiology and epidemiology, but efforts done so far had suggested consanguinity as a major risk factor for frequent mutations leading to breast cancer and has also shed light on genetic origins in different ethnic groups within Pakistan. World-wide research efforts on different ethnicities have enhanced our understanding of genetic predisposition to breast cancer but despite these discoveries, 75% of the familial risk of breast cancer remains unexplained, highlighting the fact that the majority of breast cancer susceptibility genes remain unidentified. For this purpose Pakistani population provides a strong genetic pool to elucidate the genetic etiology of breast cancer because of cousin marriages. In this review, we describe the known breast cancer predisposition factors found in the local Pakistani population and the epidemiological research work done to emphasize the importance of exploring factors/variants contributing to breast cance, in order to prevent, cure and decrease its incidence in our country.

Making personalized prostate cancer medicine a reality: challenges and opportunities in the re-establishment of gold standards.

Prostate cancer is a serious multidimensional disorder that arises because of misrepresentation of signaling cascades and acquired resistance against apoptosis. It is progressively becoming more insurmountable because of rheostat like switching of oncogenic signaling in androgen dependent and androgen depleted microenvironment. Additionally, oncogenic fusion proteins have been explored in prostate cancer tissues thus adding another layer of complexity to the targeting of protein network in cancer cell and generate hurdles in the standardization of therapy. In this Review we briefly describe identified oncogenic fusion transcripts in prostate cancer and suggest utilization of this biomarker for prostate cancer diagnosis alongwith standard PSA and immunohistochemistry analysis in Pakistan. We also provide overview of animal model studies to interpret the efficacy of vitamins.

Prevalence and molecular characterization of dengue viruses serotypes in 2010 epidemic.

During August to November 2010, a dengue (DEN) outbreak occurred, and several cities of Pakistan were affected. In this study, serum samples from 317 patients, based on clinical symptoms and blood test, were included from Lahore, Sheikhpura and Gujranwala cities for further diagnosis of different genotypes of DEN virus. The samples were categorized into 2 groups: group 1 containing patients with dengue fever (DF) without hemorrhagic manifestations (DF) (n=228) and group 2 involving patients having DF with hemorrhagic manifestations (dengue hemorrhagic fever) (n=89). The samples found positive for DEN in reverse transcriptase polymerase chain reaction (PCR) by targeting capsid and premembrane genes containing fragment of size ∼2.5 kb for groups 1 and 2 were found to be 83.33% and 91.01%, respectively; however, the detection limit was quite low by immunoglobulin M antibody capture enzyme linked immunosorbent assay in case of both groups and was found to be 34.70% and 68.53%, respectively. The 16.67% and 8.99% samples of groups 1 and 2, respectively, were found negative in reverse transcriptase PCR analysis. In case of group 1 and group 2, by adopting seminested multiplex PCR strategy, predominant genotype was DEN virus type 2 with prevalence rates of 45.17% and 65.16%, respectively. DEN virus type 1 was the second most prevailing genotype in the population with prevalence rates of 18.85% and 19.10% for groups 1 and 2, respectively.

Variable expressivity of FGF3 mutations associated with deafness and LAMM syndrome.

BACKGROUND: Recessive mutations of fibroblast growth factor 3 (FGF3) can cause LAMM syndrome (OMIM 610706), characterized by fully penetrant complete labyrinthine aplasia, microtia and microdontia. METHODS: We performed a prospective molecular genetic and clinical study of families segregating hearing loss linked to FGF3 mutations. Ten affected individuals from three large Pakistani families segregating FGF3 mutations were imaged with CT, MRI, or both to detect inner ear abnormalities. We also modeled the three dimensional structure of FGF3 to better understand the structural consequences of the three missense mutations. RESULTS: Two families segregated reported mutations (p.R104X and p.R95W) and one family segregated a novel mutation (p.R132GfsX26) of FGF3. All individuals homozygous for p.R104X or p.R132GfsX26 had fully penetrant features of LAMM syndrome. However, recessive p.R95W mutations were associated with nearly normal looking auricles and variable inner ear structural phenotypes, similar to that reported for a Somali family also segregating p.R95W. This suggests that the mild phenotype is not entirely due to genetic background. Molecular modeling result suggests a less drastic effect of p.R95W on FGF3 function compared with known missense mutations detected in fully penetrant LAMM syndrome. Since we detected significant intrafamilial variability of the inner ear structural phenotype in the family segregating p.R95W, we also sequenced FGF10 as a likely candidate for a modifier. However, we did not find any sequence variation, pointing out that a larger sample size will be needed to map and identify a modifier. We also observed a mild to moderate bilateral conductive hearing loss in three carriers of p.R95W, suggesting either a semi-dominant effect of this mutant allele of FGF3, otitis media, or a consequence of genetic background in these three family members. CONCLUSIONS: We noted a less prominent dental and external ear phenotype in association with the homozygous p.R95W. Therefore, we conclude that the manifestations of recessive FGF3 mutations range from fully penetrant LAMM syndrome to deafness with residual inner ear structures and, by extension, with minimal syndromic features, an observation with implications for cochlear implantation candidacy.