Molecular genetics, neuroimaging outcomes, and structural analyses of novel and recurrent variants of WDR62 gene in two consanguineous Pakistani families with autosomal recessive primary microcephaly.

BACKGROUND: Autosomal recessive primary microcephaly (MCPH) is a rare neurodevelopmental and genetically heterogeneous disorder, characterized by small cranium size (> - 3 SD below mean) and often results in varying degree of intellectual disability. Thirty genes have been identified for the etiology of this disorder due to its clinical and genetic heterogeneity. METHODS AND RESULTS: Here, we report two consanguineous Pakistani families affected with MCPH exhibiting mutation in WDR62 gene. The investigation approach involved Next Generation Sequencing (NGS) gene panel sequencing coupled with linkage analysis followed by validation of identified variants through automated Sanger sequencing and Barcode-Tagged (BT) sequencing. The molecular genetic analysis revealed one novel splice site variant (NM_001083961.2(WDR62):c.1372-1del) in Family A and one known exonic variant NM_001083961.2(WDR62):c.3936dup (p.Val1313Argfs*18) in Family B. Magnetic Resonance Imaging (MRI) scans were also employed to gain insights into the structural architecture of affected individuals. Neurological assessments showed the reduced gyral and sulcal patterns along with normal corpus callosum in affected individuals harboring novel variant. In silico assessments of the identified variants were conducted using different tools to confirm the pathogenicity of these variants. Through In silico analyses, both variants were identified as disease causing and protein modeling of exonic variant indicates subtle conformational alterations in prophesied protein structure. CONCLUSION: This study identifies a novel variant (c.1372-1del) and a recurrent pathogenic variant c.3936dup (p.Val1313Argfs*18) in the WDR62 gene among the Pakistani population, expanding the mutation spectrum for MCPH. These findings emphasize the importance of genetic counseling and awareness to reduce consanguinity and address the burden of this disorder.

Advances in developing metabolically engineered microbial platforms to produce fourth-generation biofuels and high-value biochemicals.

Producing bio-based chemicals is imperative to establish an eco-friendly circular bioeconomy. However, the compromised titer of these biochemicals hampers their commercial implementation. Advances in genetic engineering tools have enabled researchers to develop robust strains producing desired titers of the next-generation biofuels and biochemicals. The native and non-native pathways have been extensively engineered in various host strains via pathway reconstruction and metabolic flux redirection of lipid metabolism and central carbon metabolism to produce myriad biomolecules including alcohols, isoprenoids, hydrocarbons, fatty-acids, and their derivatives. This review has briefly covered the research efforts made during the previous decade to produce advanced biofuels and biochemicals through engineered microbial platforms along with the engineering approaches employed. The efficiency of the various techniques along with their shortcomings is also covered to provide a comprehensive overview of the progress and future directions to achieve higher titer of fourth-generation biofuels and biochemicals while keeping environmental sustainability intact.

HPLC Quantification of Thymoquinone Extracted from Nigella sativa L. (Ranunculaceae) Seeds and Antibacterial Activity of Its Extracts against Bacillus Species.

The medicinal importance of Nigella sativa seeds for treating various ailments is portrayed by its traditional uses. Owing to its immense pharmacological importance, the thymoquinone phytoconstituent of N. sativa can prove beneficial for the South Asian countries including Pakistan, where this seed is commonly produced and healthcare facilities are limited. In this study, the antibacterial activity of various extracts of N. sativa seeds, extracted thymoquinone, and oil samples have been investigated against Bacillus subtilis and Bacillus licheniformis using well and disc diffusion assay. The inhibition zones ranged between 7 and 44 mm against both the bacterial strains by well diffusion assay, while disc diffusion assay provided inhibition zones in the range of 7-23 mm. Commercial and local Kalonji oil samples were included in the study. Oil samples dissolved in methanol showed increased inhibition of bacteria. However, the extracted thymoquinone showed highest antibacterial activity. Medicine formulated using thymoquinone will prove to be an herbal alternate against the resistant microbiota associated with bacterial infections. Antibacterial activity against some Bacillus species will help signify the effect on normal gut flora when oral therapy is followed. Trying different extraction protocols can help increase extraction efficiency. Study on extraction of thymoquinone in local produce of black seed can be fruitful for conducting the stability studies and can help to gain maximum benefits from the bioactives. The crude extracts from 10 g of these seeds were subjected to preliminary phytochemical investigation. Results showed that although methanol extract had the presence of maximum phytochemicals, hexane extract was the most potent in terms of antibacterial activity. Thymoquinone, a therapeutically important bioactive in N. sativa seed, was extracted employing both solvents. TLC assay and UV spectroscopy were used for its qualitative assessment, while HPLC-UV quantification showed that 250 mg/mL of methanol extract had 368.3 µg/mL thymoquinone, while its successive extraction yielded 32.94 µg/mL thymoquinone.

Microalgal flocculation: Global research progress and prospects for algal biorefinery.

Microalgal research has made significant progress due to versatile and high-value industrial applications of microalgal biomass or its derivatives. However, to explore their full potential and to achieve commercial robustness, microalgal biorefinery needs cost-effective technologies to produce, harvest, and process the microalgal biomass on large scale as higher production and harvesting cost is one of the key hindrances in the commercialization of algae-based products. Among several other algal biomass harvesting technologies, self-flocculation seems to be an attractive, low-cost, and eco-friendly harvesting technology. This review covers various flocculation-based methods that have been employed to harvest microalgal biomass with a special emphasis on self-flocculation in microalgae. Moreover, genetic engineering approaches to induce self-flocculation in non-flocculating microalgae along with the factors affecting self-flocculation and recent research trends have also been discussed. It is concluded that self-flocculation is the most desired approach for the energy- and environment-efficient harvesting of microalgal biomass. However, its poorly understood genetic basis needs to be deciphered through detailed studies to harness its potential for the algal biorefinery.

Intrauterine growth retardation--small events, big consequences.

Intrauterine growth retardation refers to a rate of growth of a fetus that is less than normal for the growth potential of a fetus (for that particular gestational age). As one of the leading causes of perinatal mortality and morbidity, intrauterine growth retardation has immense implications for the short term and long term growth of children. It is an important public health concern in the developing countries. Health statistics encompassing parameters for maternal and child health in the Indian subcontinent have shown improvement in the past few years but they are still far from perfect. Maternal health, education and empowerment bears a strong influence on perinatal outcomes including intrauterine growth retardation and should be the primary focus of any stratagem targeted at reducing the incidence of intrauterine growth retardation. A concerted liaison of various medical and social disciplines is imperative in this regard.

Knowledge, attitudes and practices survey on organ donation among a selected adult population of Pakistan.

BACKGROUND: To determine the knowledge, attitudes and practices regarding organ donation in a selected adult population in Pakistan. METHODS: Convenience sampling was used to generate a sample of 440; 408 interviews were successfully completed and used for analysis. Data collection was carried out via a face to face interview based on a pre-tested questionnaire in selected public areas of Karachi, Pakistan. Data was analyzed using SPSS v.15 and associations were tested using the Pearson's Chi square test. Multiple logistic regression was used to find independent predictors of knowledge status and motivation of organ donation. RESULTS: Knowledge about organ donation was significantly associated with education (p = 0.000) and socioeconomic status (p = 0.038). 70/198 (35.3%) people expressed a high motivation to donate. Allowance of organ donation in religion was significantly associated with the motivation to donate (p = 0.000). Multiple logistic regression analysis revealed that higher level of education and higher socioeconomic status were significant (p < 0.05) independent predictors of knowledge status of organ donation. For motivation, multiple logistic regression revealed that higher socioeconomic status, adequate knowledge score and belief that organ donation is allowed in religion were significant (p < 0.05) independent predictors. Television emerged as the major source of information. Only 3.5% had themselves donated an organ; with only one person being an actual kidney donor. CONCLUSION: Better knowledge may ultimately translate into the act of donation. Effective measures should be taken to educate people with relevant information with the involvement of media, doctors and religious scholars.