Genomic insights into local adaptation of upland cotton in China and Pakistan.

KEY MESSAGE: Different kinship and resistance to cotton leaf curl disease (CLCuD) and heat were found between upland cotton cultivars from China and Pakistan. 175 SNPs and 82 InDels loci related to yield, fiber quality, CLCuD, and heat resistance were identified. Elite alleles found in Pakistani accessions aided local adaptation to climatic condition of two countries. Adaptation of upland cotton (Gossypium hirsutum) beyond its center of origin is expected to be driven by tailoring of the genome and genes to enhance yield and quality in new ecological niches. Here, resequencing of 456 upland cotton accessions revealed two distinct kinships according to the associated country. Fiber quality and lint percentage were consistent across kinships, but resistance to cotton leaf curl disease (CLCuD) and heat was distinctly exhibited by accessions from Pakistan, illustrating highly local adaption. A total of 175 SNP and 82 InDel loci related to yield, fiber quality, CLCuD and heat resistance were identified; among them, only two overlapped between Pakistani and Chinese accessions underscoring the divergent domestication and improvement targets in each country. Loci associated with resistance alleles to leaf curl disease and high temperature were largely found in Pakistani accessions to counter these stresses prevalent in Pakistan. These results revealed that breeding activities led to the accumulation of unique alleles and helped upland cotton become adapted to the respective climatic conditions, which will contribute to elucidating the genetic mechanisms that underlie resilience traits and help develop climate-resilient cotton cultivars for use worldwide.

Berberine: Pharmacological Features in Health, Disease and Aging.

BACKGROUND: Berberine is the main active compound of different herbs and is defined as an isoquinoline quaternary botanical alkaloid found in barks and roots of numerous plants. It exhibits a wide range of pharmacological effects, such as anti-obesity and antidiabetic effects. Berberine has antibacterial activity against a variety of microbiota, including many bacterial species, protozoa, plasmodia, fungi, and trypanosomes. OBJECTIVE: This review describes the role of berberine and its metabolic effects. It also discusses how it plays a role in glucose metabolism, fat metabolism, weight loss, how it modulates the gut microbiota, and what are its antimicrobial properties along with its potential side effects with maximal tolerable dosage. METHODS: Representative studies were considered and analyzed from different scientific databases, including PubMed and Web of Science, for the years 1982-2022. RESULTS: Literature analysis shows that berberine affects many biochemical and pharmacological pathways that theoretically yield a positive effect on health and disease. Berberine exhibits neuroprotective properties in various neurodegenerative and neuropsychological ailments. Despite its low bioavailability after oral administration, berberine is a promising tool for several disorders. A possible hypothesis would be the modulation of the gut microbiome. While the evidence concerning the aging process in humans is more limited, preliminary studies have shown positive effects in several models. CONCLUSION: Berberine could serve as a potential candidate for the treatment of several diseases. Previous literature has provided a basis for scientists to establish clinical trials in humans. However, for obesity, the evidence appears to be sufficient for hands-on use.

Drought and heat stress on cotton genotypes suggested agro-physiological and biochemical features for climate resilience.

This study aimed to investigate the impact of individual drought, heat, and combined drought and heat stress on twelve cotton genotypes, including eight tolerant and four susceptible genotypes. A field experiment was carried out by employing a randomized complete block split-plot design, with treatments (control, drought, heat, drought + heat), and cotton genotypes assigned to the main plots and sub-plots respectively. The results showed that the combined stress had a more severe impact on the yield and fiber quality of cotton genotypes compared to individual stresses. Among the studied genotypes, FB-Shaheen, FH-207, MNH-886, and White Gold exhibited superior performance in regard to agronomic and fiber quality characters under combined stress environments. Physiological parameters, including transpiration rate, stomatal conductance, relative water contents, and photosynthetic rate, were significantly reduced under combined stress. However, specific genotypes, MNH-886, FH-207, White Gold, and FB-Shaheen, demonstrated better maintenance of these parameters, indicating their enhanced tolerance to the combined stress. Furthermore, the accumulation of reactive oxygen species was more pronounced under combined stress compared to individual stressors. Tolerant genotypes showed lower levels of H2O2 and MDA accumulation, while susceptible genotypes exhibited higher levels of oxidative damage. Antioxidant enzyme activities, such as superoxide dismutase, peroxidase, and catalase, increased under combined stress, with tolerant genotypes displaying higher enzyme activities. Conversely, susceptible genotypes (AA-703, KZ 191, IR-6, and S-15) demonstrated lower increases in enzymatic activities under combined stress conditions. Biochemical traits, including proline, total phenolic content, flavonoids, and ascorbic acid, exhibited higher levels in resistant genotypes under combined stress, while sensitive genotypes displayed decreased levels of these traits. Additionally, chlorophyll a & b, and carotenoid levels were notably decreased under combined stress, with tolerant genotypes experiencing a lesser decrease compared to susceptible genotypes.

Potential of GJA8 gene variants in predicting age-related cataract: A comparison of supervised machine learning methods.

Cataracts are the problems associated with the crystallins proteins of the eye lens. Any perturbation in the conformity of these proteins results in a cataract. Age-related cataract is the most common type among all cataracts as it accounts for almost 80% of cases of senile blindness worldwide. This research study was performed to predict the role of single nucleotide polymorphisms (SNPs) of the GJA8 gene with age-related cataracts in 718 subjects (400 age-related cataract patients and 318 healthy individuals). A comparison of supervised machine learning classification algorithm including logistic regression (LR), random forest (RF) and Artificial Neural Network (ANN) were presented to predict the age-related cataracts. The results indicated that LR is the best for predicting age-related cataracts. This successfully developed model after accounting different genetic and demographic factors to predict cataracts will help in effective disease management and decision-making medical practitioner and experts.

Characterization and implication of microplastics on riverine population of the River Ravi, Lahore, Pakistan.

Microplastic (MP) pollution in the aquatic environment is an emerging subject worldwide. So far, very few investigations have been reported on the riverine fish population. This study investigated the implications of microplastics for three freshwater fish species (Labeo rohita, Cirrihinus mrigala, and Sperata seenghala) as bioindicators of this pollution. Raman spectroscopy was used to confirm MP polymer type and their distribution in water, sediments, and in different organs (gut, gills, liver, and muscles) of Labeo rohita, Cirrihinus mrigala, and Sperata seenghala collected from River Ravi at two sites (site I, Dhand Nano Dogar and site II, Jhamra). These selected sites were situated predominantly near agricultural lands and received polluted water from nearby sewerage and industries that represented potential sources of microplastic pollution. Histological analysis was combined with Raman spectroscopy to assess the effects of MPs on fish organs. MPs were identified in water and sediment samples with an average load (per 0.5 L or per 0.5 kg) of 33 items and 64 items for water and sediments at site I and 27 items and 19 items at site II, respectively. Of total MPs identified, 56.9% were found in bottom feeder C. mrigala, 37.91% in column feeder L. rohita, and 5.21% in S. seenghala at site I while at site II 60% were found in C. mrigala, 29% in L. rohita and 10.34% in S. seenghala. This was linked with more plastic accumulation in sediments from the nearby residential sewerage and industrial effluent flow. In this study, the identified MPs polymers were in the order of polyvinyl chloride (PVC) > polystyrene (PS) > propylene (PP) > polyethylene (PE). Among plastic shapes, fiber (58%) was the dominant plastic in water followed by fragment (21%), sheet (12%), and cube (9%). In sediment, the fragment was the common plastic shape with 51% followed by fiber (28%), sheet (19%), and cube (2%). Fragments (62.9%) in water and fibers (68.4%) in sediments were abundant at site 2. Microplastic mean occurrence in organs was in the order of gut > gills > muscles > liver at both sites. Significant histological alterations were observed in all three species including intestinal edema, hyperplasia, hepatocyte infiltration, accumulation of lipid droplets in the liver, lamellar fusion and breakage in gills, and muscle fiber necrosis. This study showed MP occurrence in the selected freshwater fishes, so further research is needed to assess plastic pollution in the riverine fish population of Pakistan. This study appeared to be the first in the selected area, as no significant information regarding plastic pollution in that riverine system was found when this study was conducted.

The Role of Astaxanthin as a Nutraceutical in Health and Age-Related Conditions.

The current review provides an up-to-date analysis of scientific data on astaxanthin (ASX) sources and experimental studies on its health benefits as a potent antioxidant in the aging process. ASX is a liposoluble carotenoid nutrient and reddish-orange pigment, naturally synthesized by numerous microalgae, yeasts, and bacteria as secondary metabolites. Provides a reddish hue to redfish and shellfish flesh that feed on ASX-producing microorganisms. The microalga Haematococcus pluvialis is the most important source for its industrial bioproduction. Due to its strong antioxidant properties, numerous investigations reported that natural ASX is a more significant antioxidant agent than other antioxidants, such as vitamin C, vitamin E, and ß-carotene. Furthermore, several data show that ASX possesses important nutraceutical applications and health benefits, especially in healthy aging processes. However, further studies are needed for a deeper understanding of the potential mechanisms through which ASX could lead to its effective role in the healthy aging process, such as supporting brain health and skin homeostasis. This review highlights the current investigations on the effective role of ASX in oxidative stress, aging mechanisms, skin physiology, and central nervous system functioning, and shows the potential clinical implications related to its consumption.

The Use of Wearable ECG Devices in the Clinical Setting: a Review.

Purpose of Review: This review investigates the use of wearable electrocardiograms (ECGs) in the clinic and acute care setting, and their impact on patient care, particularly pertaining to the management of cardiac arrhythmias. Recent Findings: Wearable ECGs have consistently demonstrated their non-inferiority in detecting arrhythmias when compared to the current standard of care. Different studies have highlighted their ability to improve patient care and reduce healthcare costs, while more devices are being created to work as a screening tool at a larger scale or to fit the physical abilities of a variety of patients. Summary: The use of wearable cardiac monitoring devices demonstrated considerable symptom-rhythm correlation in various clinical settings, which often resulted in a reduction in time to diagnosis and lower rates of ED visits. However, this relatively new technology raised concerns for patient accessibility and privacy among others. Further research is needed to assess their sensitivity and specificity in the clinical setting, as well as their limitations.

Comparative studies of anticancer, antimicrobial, antidiabetic, antioxidant activities of Daphne oleoides and Berberis baluchistanica extracts native to Pakistan.

To investigate in vitro anticancer, antimicrobial, antioxidant and in vivo hypoglycaemic effects of crude methanolic extracts (CMEs) of Berberis baluchistanica and Daphne oleoides. MTT assay for cytotoxicity on HeLa and NIH cells, disc diffusion protocols for antimicrobial and DPPH assay for antioxidant potential were applied. In vivo hypoglycaemic effect was investigated on Alloxan-induced diabetic rabbits. D. oleoides CME exhibited moderate cytotoxic behaviour against HeLa cells (IC50 77.87µg/mL) whereas B. baluchistanica CME was found deficient (IC50 170.02µg/mL). P. aeruginosa was susceptible to both CMEs. M. luteus and B. subtilis was prone to the bactericidal effects of D. oleoides and B. baluchistanica CMEs respectively. D. oleoides CME inhibited more than 80% S. cerevisiae and 60% C. glabrata colonies. B. baluchistanica CME showed significant antioxidant activity (IC50 52.86µg/ml) than D. oleoides CME (IC50 87.30µg/ml) and standard resveratrol (IC50 109.46µg/ml). B. baluchistanica CME showed superior antidiabetic effect (135.75 mg/dl ±0.53) as compared to D. oleoides CME (191.50 mg/dl ± 0.48) but less antidiabetic effect than metformin hydrochloride (standard). All the above potentials exhibited by D. oleoides and B. baluchistanica CMEs propose further investigations to isolate and purify responsible biologically active lead molecule(s) with diverse capabilities.

Interrelated Oncogenic Viruses and Breast Cancer.

Breast Cancer is a multifactorial disease and recent evidence that viruses have a greater role in its aetiology and pathophysiology than previously hypothesized, has garnered a lot of attention in the past couple of years. After the role of Mouse Mammary Tumour Virus (MMTV) in the oncogenesis of breast cancer has been proved in mice, search for similar viruses found quite a plausible relation of Human Papilloma Virus (HPV), Epstein-Barr virus (EBV), and Bovine Leukaemia Virus (BLV) with breast cancer. However, despite practical efforts to provide some clarity in this issue, the evidence that viruses cause breast cancer still remains inconclusive. Therefore, this article aims to clarify some ambiguity and elucidate the correlation of breast cancer and those particular viruses which are found to bring about the development of tumorigenesis by a previous infection or by their own oncogenic ability to manipulate the molecular mechanisms and bypass the immune system of the human body. Although many studies have reported, both, the individual and co-existing presence of HPV, EBV, MMTV, and BLV in patient sample tissues, particularly in Western women, and proposed oncogenic mechanisms, majority of the collective survey of literature fails to provide a delineated and strong conclusive evidence that viruses do, in fact, cause breast cancer. Measures to prevent these viral infections may curb breast cancer cases, especially in the West. More studies are needed to provide a definite conclusion.

REVIEW: Nanobiotechnology: Cradle for revolution in drug carrier systems.

The role of nanobiotechnology in the treatment of diseases is limitless. In this review we tried to focus main aspects of nanotechnology in drug carrier systems for treatment and diagnosis of various diseases such as cancer, pulmonary diseases, infectious diseases, vaccine development, diabetes mellitus and the role of nanotechnology on our economy and its positive social impacts on our community. We discussed here about the different "Biotechnano Strategies" to develop new avenues and ultimately improve the treatment of multiple diseases.

Phytochemical, acute toxicity and renal protective appraisal of Ajuga parviflora hydromethanolic leaf extract against CCl4 induced renal injury in rats.

BACKGROUND: Degenerative kidney diseases are mostly associated with oxidative stress. Natural products are considered as the antioxidants enrich food that can restrict the progress of oxidative stress induced disorders. Therefore, the present study was aimed to evaluate the renal protective effect of Ajuga parviflora leaf extract in carbon tetrachloride intoxicated rats. METHODS: The hydromethanolic extract of A. parviflora leaves was obtained by extracting twice in 60% methanol. The principal bioactive constituents were detected by LC/MS analysis. Toxicity of plant extract was assessed using brine shrimp lethal toxicity test and acute toxicity model on healthy Sprague-Dawley male rats. Nephroprotective effects of plant extract were also evaluated on rats by inducing CCl4 renal toxicity in comparison with positive control and naïve groups. The dose of A. parviflora administered to animal was 100, 200 and 300 mg/kg. All administrations were given orally on an alternate day basis for 30 days. Urine and serum biomarkers were analyzed, along with antioxidant enzymes. Finally, the DNA damages, lipid peroxides, hydrogen peroxides and nitrites were assessed in rat's renal tissue. The histopathology alterations in renal tissues were further studied for kidney damages. RESULTS: The LC/MS analysis confirmed the presence of different important pharmacological compounds in A. parviflora methanolic leaf extract. The key bioactive compounds include pyocyanin, zonisamide, D Saccharic acid, altretamine, carbocyclic thromboxane A2, Sinapyl alcohol, and vitamin C. The important polypeptides identified include Lys-Tyr-Lys, His-His-Lys, Met-Asp-Arg, Phe-Val-Arg, and PyroGlu-Val-Arg. The LD50 of A. parviflora was found to be > 1000 µg/mL. A. parviflora administration significantly subsides CCl4 toxicity in rats, reduced the elevated level of RBCs, pus and epithelial cells. The abnormal elevated level of specific gravity, creatinine, urobilinogen, urea and albumin were also reduced to normal physiological level. The reduced urinary protein and pH were also normalized. The serum urobilinogen, urea and total bilirubin levels were also reversed to normal levels while the diminished albumin and total protein levels also came to normal. The important phase I and II enzyme levels were also reversed in A. parviflora administered rats. The H2O2, thiobarbituric acid reactive substance (TBARS) and nitrite levels were significantly decreased. Furthermore, the damaged DNA and histopathological changes in CCl4 exposed rats were also highly significantly reversed after the administration of A. parviflora. All effects were significant (P < 0.05) and highly significant (P < 0.005) at 100 and 300 mg/kg respectively. CONCLUSION: The restored urine and serum profile of various parameters to normal physiological levels suggests that the A. parviflora has potential antioxidant and repairing potential in renal disorders.

Osthole: A Multifunctional Natural Compound with Potential Anticancer, Antioxidant and Anti-inflammatory Activities.

Nature has always proved to be a significant reservoir of bioactive scaffolds that have been used for the discovery of drugs since times. Medicinal plants continue to be a solid niche for biologically active and therapeutically effective chemical entities, opening up new avenues for the successful treatment of several human diseases. The contribution of plant-derived compounds to drug discovery, either in their original or in the semi-synthetic derivative form, extends far back in time. This review aims to focus on the sources, biological, and pharmacological profile of a pharmacologically active plant-derived coumarin, osthole, which is an important component of numerous remedial plants such as Cnidium monnieri. Several studies have revealed that osthole possess pharmacological properties such as anticancer, antioxidant, anti-hyperglycemic, neuroprotective, and antiplatelet. Osthole has been reported to regulate various signaling pathways, which in turn modulate several apoptosis-related proteins, cell cycle regulators, protein kinases, transcriptional factors, cytokines, and growth receptors affiliated with inflammation, proliferation and several other ailments. Osthole is known to halt proliferation and metastasis of cancerous cells by arresting the cell cycle and inducing apoptosis. The data in this review paper supports the pharmacological potential of osthole but further experimentation, biosafety profiling and synergistic effects of this compound need to be focused by the researchers to understand the full spectrum of pharmacological potential of this therapeutically potent compound.

Novel Mutations in CLPP, LARS2, CDH23, and COL4A5 Identified in Familial Cases of Prelingual Hearing Loss.

We report the underlying genetic causes of prelingual hearing loss (HL) segregating in eight large consanguineous families, ascertained from the Punjab province of Pakistan. Exome sequencing followed by segregation analysis revealed seven potentially pathogenic variants, including four novel alleles c.257G>A, c.6083A>C, c.89A>G, and c.1249A>G of CLPP, CDH23, COL4A5, and LARS2, respectively. We also identified three previously reported HL-causing variants (c.4528C>T, c.35delG, and c.1219T>C) of MYO15A, GJB2, and TMPRSS3 segregating in four families. All identified variants were either absent or had very low frequencies in the control databases. Our in silico analyses and 3-dimensional (3D) molecular modeling support the deleterious impact of these variants on the encoded proteins. Variants identified in MYO15A, GJB2, TMPRSS3, and CDH23 were classified as "pathogenic" or "likely pathogenic", while the variants in CLPP and LARS2 fall in the category of "uncertain significance" based on the American College of Medical Genetics and Genomics/Association for Molecular Pathology (ACMG/AMP) variant pathogenicity guidelines. This paper highlights the genetic diversity of hearing disorders in the Pakistani population and reports the identification of four novel mutations in four HL families.

Cardamonin: A new player to fight cancer via multiple cancer signaling pathways.

Nature's pharmacy has undoubtedly served humans as an affordable and safer health-care regime for a long times. Cardamonin, a chalconoid present in several plants has been known for a longtime to have beneficial properties towards human health. In this review, we aimed to highlight the recent advances achieved in discovering the pharmacological properties of cardamonin. Cardamonin is cardamom-derived chalcone, which plays a role in cancer treatment, immune system modulation, inflammation and pathogens killing. Through the modulation of cellular signaling pathways, cardamonin activates cell death signal to induce apoptosis in malignant cells that results in the inhibition of cancer development. Moreover, cardamonin arrests cell cycle by altering the expression of regulatory proteins during malignant cells division. Due to its relatively selective cytotoxic potential against host malignant cells, cardamonin is emerging as a promising novel experimental anticancer agent. The potential of cardamonin to target various signaling molecules, transcriptional factors, cytokines and enzymes, such as mTOR, NF-κB, Akt, STAT3, Wnt/ß-catenin and COX-2 enhances the opportunity to explore it as a new multi-target therapeutic agent. The pharmacokinetic and biosafety profile of cardamonin favor it as a potentially safe biomolecule for pharmaceutical drug development.

Biochanin A: A novel bioactive multifunctional compound from nature.

Natural products (NPs) will continue to serve humans as matchless source of novel drug leads and an inspiration for the synthesis of non-natural drugs. As our scientific understanding of 'nature' is rapidly expanding, it would be worthwhile to illuminate the pharmacological distinctions of NPs to the scientific community and the public. Flavonoids have long fascinated scientists with their remarkable structural diversity as well as biological functions. Consequently, this review aims to shed light on the sources and pharmacological significance of a dietary isoflavone, biochanin A, which has been recently emerged as a multitargeted and multifunctional guardian of human health. Biochanin A possesses anti-inflammatory, anticancer, neuroprotective, antioxidant, anti-microbial, and hepatoprotective properties. It combats cancer development by inducing apoptosis, inhibition of metastasis and arresting cell cycle via targeting several deregulated signaling pathways of cancer. It fights inflammation by blocking the expression and activity of pro-inflammatory cytokines via modulation of NF-κB and MAPKs. Biochanin A acts as a neuroprotective agent by inhibiting microglial activation and apoptosis of neurons. As biochanin A has potential to modulate several biological networks, thus, it can be anticipated that this therapeutically potent compound might serve as a novel lead for drug development in the near future.

Comparative study of antibacterial activity between Schiff base nicotinic hydrazide derivative and its silver architected nanoparticles with atomic force microscopic study of bacterial cell wall.

The threat of multi-drug resistant bacterial pathogens evokes researchers to synthesized safe and effective chemotherapeutic agents for nano-drug delivery system. In current study, Schiff base of nicotinic hydrazide(NHD) and its silver nanoparticles(NHD-AgNPs) were synthesized and characterized. These compounds were investigated for cytotoxicity, antibacterial and AFM activity. The NHD showed LD50 at >1000µg/mL while NHD-AgNPs didn't exhibit toxicity at 1000µg/mL against 3T3 cell line. The NHD showed zone of inhibition against two strains of salmonella enteric (ATCC 14028 and 700408) 45.29±1.66 and 48.01±1.43mm respectively at 160µg/mL (p<0.01) while NHD-AgNPs exhibited 55.87±2.08 and 52.88±1.42 mm respectively at 130µg/mL (p<0.001) in disc diffusion method. NHD showed more than 70% growth inhibition for both strains at 85 and 125µg/ml (p<0.01) respectively, while NHD-AgNPs inhibit 80% and 75% respectively at 75 and 125 µg/ml (p<0.01, p<0.001) against Alamar blue antibacterial assay. For morphological changes in bacterial cell wall NHD and NHD-AgNPs treated bacterial cells were observed under atomic force microscope(AFM) and treated bacterial cells were severely damaged with leaked cytoplasmic contents as compare to untreated bacterial cell. These results validate that NHD-AgNPs were highly active as compared to NHD against both strains at their MIC concentrations. In future, comparative wound healing potential will be emphasized.

In vitro cytotoxic, antioxidant, antibacterial and antifungal activity of Saussurea heteromalla indigenous to Pakistan.

Medicinal plants are proven to reveal vast promising potential providing novel drug candidates to combat health-related problems. The aim of current study is to discover new drug compounds with anti-anticancer, antioxidant, antibacterial and antifungal potential, to serve the purpose Saussurea heteromalla (Family: Asteraceae) indigenous to Pakistan was screened for the in vitro cytotoxicity against HeLa cells (Human cervical cancer cell line) compared to the NIH / 3T3 cells (mouse normal fibroblast cells) by performing the MTT colorimetric assay and antifungal, antibacterial and antioxidant potential by adopting standard protocols. S. heteromalla crude methanolic extract (CME) demonstrated strong cytotoxic potential against HeLa cells at 200µg/mL; (77.28 ±1.53% kill; IC50: 62.13µg/mL) compared to standard doxorubicin (95.90% kill; IC50: 0.2µg/mL). Inhibitory Zone of the extract at concentrations (30, 60, 90µg/mL) against Bacillus subtilis, Serratia marecescens, Staphylococcus aureus, Micrococcus luteus, Pseudomonas aeruginosa, Strptotropomonas maltophilia, Escherichia coli, Salmonella typhi, and Saccharomyces cerevisiae, Candida albicans, Candida glabrata was measured. Manifestation of intensified results against Gram-negative Serratia marecescens qualifies the S. heteromalla extract as a considerable source of narrow spectrum antibiotic. However, antifungal activity against C. albicans was found to be logical. Antioxidant potential was determined through DPPH assay which declared no notable antioxidant effects. To the best of our knowledge this is first research and report on above mentioned biological studies of S. heteromalla.

Global genetic insight contributed by consanguineous Pakistani families segregating hearing loss.

Consanguineous Pakistani pedigrees segregating deafness have contributed decisively to the discovery of 31 of the 68 genes associated with nonsyndromic autosomal recessive hearing loss (HL) worldwide. In this study, we utilized genome-wide genotyping, Sanger and exome sequencing to identify 163 DNA variants in 41 previously reported HL genes segregating in 321 Pakistani families. Of these, 70 (42.9%) variants identified in 29 genes are novel. As expected from genetic studies of disorders segregating in consanguineous families, the majority of affected individuals (94.4%) are homozygous for HL-associated variants, with the other variants being compound heterozygotes. The five most common HL genes in the Pakistani population are SLC26A4, MYO7A, GJB2, CIB2 and HGF, respectively. Our study provides a profile of the genetic etiology of HL in Pakistani families, which will allow for the development of more efficient genetic diagnostic tools, aid in accurate genetic counseling, and guide application of future gene-based therapies. These findings are also valuable in interpreting pathogenicity of variants that are potentially associated with HL in individuals of all ancestries. The Pakistani population, and its infrastructure for studying human genetics, will continue to be valuable to gene discovery for HL and other inherited disorders.