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Abstract Background/Objectives This study aims to elucidate the genetic causes of congenital hypogonadotropic hypogonadism (CHH), a rare genetic disorder resulting in GnRH deficiency, in six families from Pakistan. Methods Eighteen DNA samples from six families underwent genome sequencing followed by standard evaluation for pathogenic single nucleotide variants (SNVs) and small indels. All families were subsequently analyzed for pathogenic copy number variants (CNVs) using CoverageMaster. Results Novel pathogenic homozygous SNVs in known CHH genes were identified in four families: two families with variants in GNRHR, and two others harboring KISS1R variants. Subsequent investigation of CNVs in the remaining two families identified novel unique large deletions in ANOS1. Conclusion A combined, systematic analysis of single nucleotide and CNVs helps to improve the diagnostic yield for variants in patients with CHH.

ObjectiveThis study aims to clinically and genetically assess 30 unrelated consanguineous Pakistani families from various ethnic backgrounds, all exhibiting features of neurodevelopmental disorders (NDDs).MethodsWe conducted clinical, genetic, biochemical, and molecular analyses on 30 consanguineous families with NDDs enrolled from various regions of Pakistan. The likely molecular causes of primary microcephaly and NDDs were identified. Detailed clinical investigations and molecular diagnoses were performed using whole exome sequencing (WES) of the proband, followed by Sanger sequencing for validation and segregation in the available family members of the affected families.ResultsWES identified likely disease-causing homozygous variants in 30 unrelated consanguineous families. Six families presented newly described variants in known NDD-related genes: ABAT (c.1439 T > G; p.Phe480Cys) [OMIM613163], SLC12A6 (c.2865_2865insT; p.Glu955Asnfs*5) [OMIM 218000], SHANK3 (c.1305-3_1,305-2delTT; p.Gln29-_Gly305del) [OMIM 606232], BCKDK (c.356_356insC; p.Gly119Alafs*24) [OMIM 614923], DDHD2 (c.2065G > T; p.Asp689Tyr) [OMIM 615033], ERCC2 (c.1255G > A; p.Glu419Lys) [OMIM 610756]. Additionally, 12 families had previously reported disease-causing variants associated with different types of NDDs: ATRX (c.109C > T; p.Arg37*) [OMIM 309580], GPR56 [ADGRG1] (c.1423C > T; p.Arg475*) [OMIM 606854], NAGLU (c.1694G > A; p.Arg565Gln) [OMIM 252920], DOLK (c.3G > A; p.Met1Ile) [OMIM 610768], GPT2 (c.815C > T; p.Ser272Leu) [OMIM 616281], DYNC1I2 (c.607 + 1G > A; p.?) [OMIM 618492], FBXL3 (c.885delT; p.Leu295Phefs25*) [OMIM 606220], LINGO1 (c.869G > A; p.Arg290His) [OMIM 618103], and ASPM (c.3978G > A; Trp1326*, c.9557C > G; p.Ser3186*, c.6994C > T; p.Arg2332*) [OMIM 608716]. All the identified variants showed segregation compatible with autosomal recessive inheritance.ConclusionIn the present study, we observed a high frequency of ASPM variants in the genetic analysis of 30 consanguineous families exhibiting features of NDDs, particularly those associated with autosomal recessive primary microcephaly. These findings contribute to studies on genotype–phenotype correlation, genetic counseling for families, and a deeper understanding of human brain function and development.

Cohen Syndrome (CS) is a rare autosomal recessive disorder caused by biallelic mutations in the VPS13B gene. It is characterized by multiple clinical features, including acquired microcephaly, developmental delay, intellectual disability, neutropenia, and retinal degeneration. VPS13B is part of the bridge-like lipid transport (BLTP) protein family, which in mammals also includes VPS13A, -C, and -D. The proteins of this family are peripheral membrane proteins with different sub-cellular localization, but all share similar structural features and have been proposed to act as lipid transport proteins at organellar membrane contact sites. VPS13B is localized at the Golgi apparatus and is essential for the maintenance of organelle architecture. Here we present a review of the experimental data on the function of the protein at the cellular level, discussing the potential link with disease phenotype and review the studies on animal models recapitulating features of the human disease.

Abstract Craniofacial microsomia (CFM; also known as Goldenhar syndrome), is a craniofacial developmental disorder of variable expressivity and severity with a recognizable set of abnormalities. These birth defects are associated with structures derived from the first and second pharyngeal arches, can occur unilaterally and include ear dysplasia, microtia, preauricular tags and pits, facial asymmetry and other malformations. The inheritance pattern is controversial, and the molecular etiology of this syndrome is largely unknown. A total of 670 patients belonging to unrelated pedigrees with European and Chinese ancestry with CFM, are investigated. We identify 18 likely pathogenic variants in 21 probands (3.1%) in FOXI3. Biochemical experiments on transcriptional activity and subcellular localization of the likely pathogenic FOXI3 variants, and knock-in mouse studies strongly support the involvement of FOXI3 in CFM. Our findings indicate autosomal dominant inheritance with reduced penetrance, and/or autosomal recessive inheritance. The phenotypic expression of the FOXI3 variants is variable. The penetrance of the likely pathogenic variants in the seemingly dominant form is reduced, since a considerable number of such variants in affected individuals were inherited from non-affected parents. Here we provide suggestive evidence that common variation in the FOXI3 allele in trans with the pathogenic variant could modify the phenotypic severity and accounts for the incomplete penetrance.

Abstract Seed shattering is a critical challenge that significantly reduces sesame production by 50%. These shattering losses can be reduced by selecting shattering resistant genotypes or by incorporating modern agronomic management such as paclobutrazol, which can boost productivity and prevent seed shattering in sesame. Two-years of field trials were conducted to examine the effect of sesame genotypes, environment, and paclobutrazol (PBZ) concentrations. Twelve sesame genotypes were used in a four-way factorial RCBD with three replications and five PBZ concentrations (T0 = Control; T1 = 150; T2 = 300; T3 = 450; and T4 = 600 mg L− 1) under rainfed conditions of Pothwar. The findings revealed significant variations in the major effects of all examined variables (genotypes, locations, years, and PBZ levels). Sesame genotypes PI-154304 and PI-175907 had the highest plant height, number of capsule plant− 1, seed capsule− 1, 1000 seed weight, biological yield, and seed yield, while also having the lowest seed losses and shattering percentage. Regarding environments, NARC-Islamabad generated the highest plant height, number of capsule plant− 1, shattering percentage, and biological yield; however, the URF-Koont produced the highest seed yield with the lowest shattering percentage. Additionally, plant height, capsules plant− 1, and biological yield were higher in 2021, while seed capsule− 1, 1000 seed weight, seed losses, shattering percentage, and seed yield were higher in 2020. PBZ concentration affected all measured parameters; plant height and number of seed capsule− 1 decreased with increasing PBZ concentrations. 450 mg L− 1 PBZ concentration generated the highest biomass, number of capsules plant− 1, and seed yield. At the same time, PBZ concentration 600 mg L− 1 generated the smallest plant, the lowest seed capsules− 1, the greatest thousand seed weight, and the lowest shattering percentage. The study concluded that paclobutrazol could dramatically reduce shattering percentage and shattering losses while increasing economic returns through better productivity. Based on the findings, the genotypes PI-154304 and PI-175907 with paclobutrazol level 450 mgL− 1 may be suggested for cultivation in Pothwar farming community under rainfed conditions, as they showed promising shattering resistance as well as enhanced growth and yield.

The vacuolar protein sorting-associated protein 13B (VPS13B) is a large and highly conserved protein. Disruption of VPS13B causes the autosomal recessive Cohen syndrome, a rare disorder characterized by microcephaly and intellectual disability among other features, including developmental delay, hypotonia, and friendly-personality. However, the underlying mechanisms by which VPS13B disruption leads to brain dysfunction still remain unexplained.To gain insights into the neuropathogenesis of Cohen syndrome, we systematically characterized brain changes in Vps13b-mutant mice and compared murine findings to 235 previously published and 17 new patients diagnosed with VPS13B-related Cohen syndrome.We showed that Vps13b is differentially expressed across brain regions with the highest expression in the cerebellum, the hippocampus and the cortex with postnatal peak. Half of the Vps13b−/− mice die during the first week of life. The remaining mice have a normal lifespan and display the core phenotypes of the human disease, including microcephaly, growth delay, hypotonia, altered memory, and enhanced sociability. Systematic 2D and 3D brain histo-morphological analyses reveal specific structural changes in the brain starting after birth. The dentate gyrus is the brain region with the most prominent reduction in size, while the motor cortex is specifically thinner in layer VI. The fornix, the fasciculus retroflexus, and the cingulate cortex remain unaffected. Interestingly, these neuroanatomical changes implicate an increase of neuronal death during infantile stages with no progression in adulthood suggesting that VPS13B promotes neuronal survival early in life. Importantly, whilst both sexes were affected, some neuroanatomical and behavioral phenotypes were less pronounced or even absent in females. We evaluate sex differences in Cohen patients and conclude that females are less affected both in mice and patients.Our findings provide new insights about the neurobiology of VPS13B and highlight previously unreported brain phenotypes while defining Cohen syndrome as a likely new entity of non-progressive infantile neurodegeneration.

The excessive use of chemical fertilizers is deteriorating both the environment and soil, making it a big challenge faced by sustainable agriculture. To assist the efforts for the solution of this burning issue, nine different potential native strains of plant growth-promoting bacteria (PGPB) namely, SA-1(Bacillus subtilis), SA-5 (Stenotrophomonas humi),SA-7(Azospirillum brasilense), BH-1(Azospirillum oryzae), BH-7(Azotobacter armeniacus), BH-8(Rhizobium pusense), BA-3(Azospirillum zeae), BA-6(Rhizobium pusense), and BA-7(Pseudomonas fragi) were isolated that were characterized morphologically, biochemically and molecularly on the basis of 16S rRNA sequencing. Furthermore, the capability of indigenous PGPB in wheat (Triticum aestivum, Chakwal-50) under control, DAP+FYM, SA-1,5,7, BH-1,7,8, BA-3,6,7, DAP+ FYM + SA-1,5,7, DAP+FYM+ BH-1,7,8 and DAP+FYM+ BA-3,6,7 treatments was assessed in a randomized complete block design (RCBD). The results of the study showed that there was a significant increase in plant growth, nutrients, quality parameters, crop yield, and soil nutrients at three depths under SA-1,5,7, BH-1,7,8, and BA-3,6,7 in combination with DAP+FYM. Out of all these treatments, DAP+ FYM + BA-3,6,7 was found to be the most efficient for wheat growth having the highest 1000-grain weight of 55.1 g. The highest values for plant height, no. of grains/spike, spike length, shoot length, root length, shoot dry weight, root dry weight, 1000 grain weight, biological yield, and economic yield were found to be 90.7 cm, 87.7 cm, 7.20 cm, 53.5 cm, 33.5 cm, 4.87 g, 1.32 g, 55.1 g, 8209 kg/h, and 4572 kg/h, respectively, in the DAP+FYM+BA treatment. The DAP+FYM+BA treatment had the highest values of TN (1.68 µg/mL), P (0.38%), and K (1.33%). Likewise, the value of mean protein (10.5%), carbohydrate (75%), lipid (2.5%), and available P (4.68 ppm) was also highest in the DAP+FYM+BA combination. C:P was found to be significantly highest (20.7) in BA alone but was significantly lowest (11.9) in DAP+FYM+BA. Hence, the integration of strains BA-3, BA-5, and BA-7 in fertilizers can be regarded as the most suitable choice for agricultural growth in the sub-mountainous lower region of AJK. This could serve as the best choice for sustainable wheat growth and improved soil fertility with lesser impacts on the environment.

Abstract TET3 encodes an essential dioxygenase involved in epigenetic regulation through DNA demethylation. TET3 deficiency, or Beck-Fahrner syndrome (BEFAHRS; MIM: 618798), is a recently described neurodevelopmental disorder of the DNA demethylation machinery with a nonspecific phenotype resembling other chromatin-modifying disorders, but inconsistent variant types and inheritance patterns pose diagnostic challenges. Given TET3’s direct role in regulating 5-methylcytosine and recent identification of syndrome-specific DNA methylation profiles, we analyzed genome-wide DNA methylation in whole blood of TET3-deficient individuals and identified an episignature that distinguishes affected and unaffected individuals and those with mono-allelic and bi-allelic pathogenic variants. Validation and testing of the episignature correctly categorized known TET3 variants and determined pathogenicity of variants of uncertain significance. Clinical utility was demonstrated when the episignature alone identified an affected individual from over 1000 undiagnosed cases and was confirmed upon distinguishing TET3-deficient individuals from those with 46 other disorders. The TET3-deficient signature - and the signature resulting from activating mutations in DNMT1 which normally opposes TET3 - are characterized by hypermethylation, which for BEFAHRS involves CpG sites that may be biologically relevant. This work expands the role of epi-phenotyping in molecular diagnosis and reveals genome-wide DNA methylation profiling as a quantitative, functional readout for characterization of this new biochemical category of disease.

Abstract Intellectual disability (ID) is a highly heterogeneous disorder with hundreds of associated genes. Despite progress in the identification of the genetic causes of ID following the introduction of high-throughput sequencing, about half of affected individuals still remain without a molecular diagnosis. Consanguineous families with affected individuals provide a unique opportunity to identify novel recessive causative genes. In this report, we describe a novel autosomal recessive neurodevelopmental disorder. We identified two consanguineous families with homozygous variants predicted to alter the splicing of ATP9A which encodes a transmembrane lipid flippase of the class II P4-ATPases. The three individuals homozygous for these putatively truncating variants presented with severe ID, motor and speech impairment, and behavioral anomalies. Consistent with a causative role of ATP9A in these patients, a previously described Atp9a−/− mouse model showed behavioral changes.

OBJECTIVE: To identify the disease-causing mutation in a family with autosomal recessive primary microcephaly (MCPH). METHODOLOGY: This cross-sectional study was the continuation of an ongoing family-based study initiated in 2016 at the Department of Biochemistry, Quaid-e-Azam University, Islamabad. The family was selected randomly and recruited from Sahiwal and has three members with MCPH. DNA was isolated from blood samples and the genome-wide scan was performed to map homozygous regions. Whole exome sequencing (WES) was performed to identify the plausible gene variant. RESULTS: Whole genome data analysis identified multiple homozygous regions, but none of these contain known MCPH genes. Whole exome sequencing (WES) data identified six potentially pathogenic variants but only the Laminin subunit alpha-3 (LAMA3) (c.5260C/T) variant segregates in the family and is also present within the genomic region mapped on chromosome 18. The reevaluation of affected members of the family revealed the presence of blisters on their hands and feet indicating the presence of epidermolysis bullosa along with microcephaly. CONCLUSION: The casual finding of the LAMA3 variant (c.5260C/T; p. Arg1754Trp) and absence of any other MCPH causing variant in affected members of this family expands the phenotypic spectrum of LAMA3 associated phenotype. Therefore, we can conclude that the LAMA3 variant can probably cause recessive microcephaly and epidermolysis bullosa, but additional studies are needed to establish the role of LAMA3 in microcephaly.

The increasing contamination of soil with arsenic (As), and salinity has become a menace to food security and human health. The current study investigates the comparative efficacy of plain biochar (BC), and silicon-nanoparticles doped biochar (SBC) for ameliorating the As and salinity-induced phytotoxicity in quinoa (Chenopodium quinoa Willd.) and associated human health risks. Quinoa was grown on normal and saline soils (ECe 12.4 dS m−1) contaminated with As (0, 20 mg kg−1) and supplemented with 1% of BC or SBC. The results demonstrated that plant growth, grain yield, chlorophyll contents, and stomatal conductance of quinoa were decreased by 62, 44, 48, and 66%, respectively under the blended stress of As and salinity as compared to control. Contrary to this, the addition of BC to As-contaminated saline soil caused a 31 and 25% increase in plant biomass and grain yield. However, these attributes were increased by 45 and 38% with the addition of SBC. The H2O2 and TBARS contents were enhanced by 5 and 10-fold, respectively under the combined stress of As and salinity. The SBC proved to be more efficient than BC in decreasing oxidative stress through overexpressing of antioxidant enzymes. The activities of superoxide dismutase, peroxidase, and catalase were enhanced by 5.4, 4.6, and 11-fold with the addition of SBC in As-contaminated saline soil. Contamination of grains by As revealed both the non-carcinogenic and carcinogenic risks to human health, however, these effects were minimized with the addition of SBC. As accumulation in grains was decreased by 65-fold and 25-fold, respectively for BC and SBC in addition to As-contaminated saline soil. The addition of SBC to saline soils contaminated with As for quinoa cultivation is an effective approach for decreasing the food chain contamination and improving food security. However, more research is warranted for the field evaluation of the effectiveness of SBC in abating As uptake in other food crops cultivated on As polluted normal and salt-affected soils.

Abstract Background Intellectual disability (ID) is a phenotypically and genetically heterogeneous disorder. Methods In this study, genome wide SNP microarray and whole exome sequencing are used for the variant identification in eight Pakistani families with ID. Beside ID, most of the affected individuals had speech delay, facial dysmorphism and impaired cognitive abilities. Repetitive behavior was observed in MRID143, while seizures were reported in affected individuals belonging to MRID137 and MRID175. Results In two families (MRID137b and MRID175), we identified variants in the genes CCS and ELFN1, which have not previously been reported to cause ID. In four families, variants were identified in ARX, C5orf42, GNE and METTL4. A copy number variation (CNV) was identified in IL1RAPL1 gene in MRID165. Conclusion These findings expand the existing knowledge of variants and genes implicated in autosomal recessive and X linked ID.

Abstract Cohen syndrome (CS) is a rare syndromic form of rod-cone dystrophy. Recent case reports have suggested that cystoid maculopathy (CM) could affect CS patients with an early onset and high prevalence. Our study aims at improving our understanding and management of CM in CS patients through a retrospective case series of ten CS patients with identified pathogenic variants in VPS13B. Longitudinal optical coherence tomography (OCT) imaging was performed and treatment with carbonic anhydrase inhibitors (CAI) was provided to reduce the volume of cystoid spaces. CM affected eight out of ten patients in our cohort. The youngest patient showed a strong progression of macular cysts from the age of 4.5 to 5 years despite oral CAI medication. Other teenage and young adult patients showed stable macular cysts with and without treatment. One patient showed a moderate decrease of cystoid spaces in the absence of treatment at 22 years of age. Through a correlative analysis we found that the volume of cystoid spaces was positively correlated to the thickness of peripheral and macular photoreceptor-related layers. This study suggests that CAI treatments may not suffice to improve CM in CS patients, and that CM may resolve spontaneously during adulthood as photoreceptor dystrophy progresses.

This study investigates the effect of female chief executive officers (FCEOs) on investment efficiency. Our study suggests that CEO gender plays a significant role in efficient investment decisions by improving governance and disciplining the management. We document that FCEOs are associated with higher investment efficiency. We also find that FCEOs, when making investment decisions, pay more attention to curbing overinvestment than to underinvestment. Furthermore, we find that FCEOs play no role in improving the investment efficiency of state-owned enterprises (SOEs). Our findings suggest that SOEs’ investment decisions are independent of factors that shape investment decisions at non-state-owned enterprises and are more reliant on sociopolitical factors.

Crop irrigation with untreated wastewater is a routine practice in developing countries that causes multiple human health consequences. A comparative study was performed to regulate total Cr and Pb stress in soil and Solanum melongena L. plant. For this purpose, 0.2% chitosan polymerized silica gel (CP-silica gel), 1.5% zinc-enriched biochar (ZnBc), and three bacterial species such as Trichococcus sp. (B1), Pseudomonas alcaligenes (B2), and Bacillus subtilis (B3) were selected. Initially, a biosorption trial was conducted to test the heavy metal removal efficiency of three bacterial species B1, B2, and B3 for 24 h. Hence, B3 showed maximum Cr and Pb removal efficiency among the studied bacterial isolates. Then, a pot study was conducted with 12 different treatments having three replicates. After harvesting, different growth and biochemical parameters such as chlorophyll concentration, proteins, phenolics, reactive oxygen species, and antioxidant enzymes were analyzed. The results demonstrated that wastewater application significantly (p ≤ 0.01) reduced the fresh and dry weights of the root, stem, and leaves due to high total Cr and Pb toxicity. However, CP-silica gel and ZnBc treatments performed best when applied in combination with B3. The concentration of leaf total Cr was significantly decreased (91 and 85%) with the application of ZnBc + B3 and CP-Silica gel + B3, respectively, as compared to control. There was a reduction in stem hydrogen peroxide (87%) and malondialdehyde (81%) recorded with CP-silica gel + B3 treatment due to enhanced activities of antioxidant enzymes viz. ascorbate peroxidase (6-folds) and catalase (7-folds) relative to control. Similarly, leaf total phenolics (3-folds) and protein (6-folds) contents were enhanced with CP silica gel+B3 application relative to control. Overall, CP-silica gel and ZnBc with B3 application proved to be the most appropriate treatments and can be used in developing countries to limit the deleterious effects of total Cr and Pb pollution.

Under natural conditions, plants constantly encounter various biotic and abiotic factors, which can potentially restrict plant growth and development and even limit crop productivity. Among various abiotic factors affecting plant photosynthesis, light serves as an important factor that drives carbon metabolism in plants and supports life on earth. The two components of light (light quality and light intensity) greatly affect plant photosynthesis and other plant’s morphological, physiological and biochemical parameters. The response of plants to different spectral radiations and intensities differs in various species and also depends on growing conditions. To date, much research has been conducted regarding how different spectral radiations of varying intensity can affect plant growth and development. This review is an effort to briefly summarize the available information on the effects of light components on various plant parameters such as stem and leaf morphology and anatomy, stomatal development, photosynthetic apparatus, pigment composition, reactive oxygen species (ROS) production, antioxidants, and hormone production.

Objective(s): Lapachone is a natural naphthoquinone-derived compound found in Tabebuia avellanedae. It is well-known for its analgesic, anti-inflammatory, anti-microbial, diuretic, and anti-cancerous effects. However, the wound-healing effects of this compound are not known yet. The aim of this study was to investigate the wound healing activity of naphthoquinones (α-lapachone and β-lapachone) from Handroanthus impetiginosus. Materials and Methods: Expression of Sirt3, migration-related proteins (Rac1, Cdc42, α-Pak) and angiogenesis-related protein of vascular endothelial growth factor (VEGF) was monitored using western blot analysis. Blood vessel formation and tissue development were monitored by angiogenesis assay and hematoxylin & eosin (H & E) staining, respectively on mouse skin tissue samples. Both α-lapachone and β-lapachone increased Sirt3 expression in vivo, but only β-lapachone increased Sirt3 expression in vitro. Results: Both the compounds accelerated wound healing in cultured skin cells as well as mouse skin; however, β-lapachone was more effective at lower concentrations. Both of the compounds increased the expression of migration-related proteins both in vitro and in vivo. Similarly, α-lapachone and β-lapachone increased VEGF expression, tissue development and blood vessel formation in mouse skin.Conclusion: These findings indicated that α-lapachone and β-lapachone are novel Sirt3 activators, and Sirt3 has a role in wound healing. Thus, Sirt3 and its regulators come out as a novel target and potential drug candidates, respectively in the important field of cutaneous wound healing.

Strigolactones, a new group of phytohormones, are reported to improve plant tolerance to multiple abiotic stresses. A pot experiment was conducted to investigate the impact of synthetic strigolactone (GR24 at 0.001, 0.01 and 0.1 mg L−1) application on ornamental sunflowers (Helianthus annuus cv. Vincent’s Choice) grown under salt stress (150 mM NaCl). Salt stress was applied after 14 days, and SL was applied 25 days seed sowing. The results showed that amongst various GR24 concentrations, 0.01 mg L−1 proved to be superior, as it enhanced the photosynthetic rate (9.29%), transpiration rate (0.76%), stomatal conductance (77.5%), total soluble protein (0.55%) and K+ (14.63% in roots; 14.87% in shoots) and Ca2+ (12.63% in roots; 11.48% in shoots) contents under control conditions. Similarly, the leaf turgor potential (Ψp), osmotic potential (Ψs) and free proline, glycinebetaine (GB), superoxide dismutase (SOD), catalase (CAT) and peroxide (POD) contents increased by 58.17, 89.95, 159.04, 101.54, 74.42, 175.68 and 53.62%, respectively, under salt stress conditions. The leaf water potential (Ψw) decreased (−0.14%) and the malondialdehyde (MDA) content increased (16.65%) when treated with the 0.001 mg L−1 GR24 level. Meanwhile, hydrogen peroxide (H2O2) and Na+ concentrations in roots and shoots increased by 62.53%, 74.66% and 38.55% under saline conditions with a GR24 level of 0 mg L−1. Regarding the plant biomass, a GR24 level of 0.01 mg L−1 with salt stress greatly decreased the root (−47.27% and −50.45%) and shoot (−44.79% and −59.42%) fresh and dry weights, respectively, compared to control conditions. These results reveal that exogenously applied GR24 might be an effective way to mitigate the perilous impacts of salt stress in ornamental sunflower production. It is suggested that the use of molecular techniques to study different processes in which GR24 could play a vital part in various commercial floricultural crops is extremely imperative and can open novel horizons for future investigations in this exhilarating field of plant hormones.

Deterioration of asphalt pavements due to massive load of vehicles and climatic variation has demanded the use of pavements construction material with an excellent resilience characteristic, resistance to permanent deformation, and most importantly, a much longer service lifespan. The main structural distresses in pavement construction are permanent deformation at high temperatures and fatigue cracking under repetitive traffic loadings. To comprehensively investigate the performance of bitumen penetration grade (PG) 70 against rutting, fatigue, and high temperature cracking in hot mix asphalt (HMA) pavements, polycarbonate (PC) and polytetrafluoroethylene (PTFE) were used. The investigation of the internal structure, rheological, and physical properties of base and modified bitumen (MB) mixes with different percentages of modifiers (0%, 2.5%, and 5%) by weight were performed via scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR) analysis, X-ray diffraction (XRD) pattern analysis, rolling thin-film oven test (RTFOT), pressurized aging vessel (PAV), dynamic shear rheometer (DSR), rotational viscosity (RV), and bending beam rheometer (BBR). The results of the RV test indicate that modification of neat bitumen with polycarbonate and polytetrafluoroethylene increased the viscosity for polycarbonate-modified bitumen (PCMB), polytetrafluoroethylene-modified bitumen (PTFEMB), and for a blend of PCMB-PTFEMB by 44%, 50%, and 55.75% at 135 °C and 111.10%, 127.80%, and 138.88% at 165 °C, accordingly. BBR test results revealed that modifiers increased the rigidity of neat bitumen by 74.8%, 75.8%, and 74.5% at −16 °C, −22 °C, and −28 °C, respectively.

We present a rare case of transient central diabetes insipidus occurring in the setting of endoscopic sinus surgery, without evidence of structural intracranial/pituitary complications. Other etiologic factors were excluded. Literature review is presented about prior reported cases of diabetes insipidus associated with non-intracranial surgeries. Keywords: Sinus surgery, Maxillary antrostomy, Ethmoidectomy, Septoplasty, Diabetes insipidus

Split hand/split foot malformation (SHFM) or ectrodactyly is characterized by a deep median cleft of the hand or foot, hypoplasia or aplasia of the metacarpals, metatarsals, and phalanges. It is a clinically and genetically heterogeneous group of limb malformations. This study aimed to identify the pathogenic variant in a consanguineous Pakistani family with autosomal recessive SHFM. Peripheral blood samples were obtained, DNA was extracted, WNT10B coding and noncoding regions were PCR amplified and Sanger sequencing was performed using workflow suggested by Thermo Fisher Scientific. A novel homozygous nonsense variant (c.1098C>A; p.Cys366*) was identified in the WNT10B gene in the index patients, which probably explains SHFM type 6 in this family in comparison with similar data from the literature.

Salinity limits the growth and nutrient uptake in crop species. Studies show that both potassium (K) and humic acid (HA) improved plant tolerance to salinity. However, the interactive effect of K and HA on plant tolerance to salinity stress remains unknown. This pot study examined the effect of application of K (0, 5 or 10 mM) and HA (0 or 2 g kg−1), alone or in combination, on the growth and physiology under salinity (100 mM NaCl) in two wheat genotypes (SARC 1, salt tolerant; and SARC 5, salt sensitive). The results revealed that salt stress reduced shoot biomass by 35% and 49% in SARC 1 and SARC 5, respectively. Salinity induced overproduction of H2O2 and lipid peroxidation in both genotypes, but the decline in pigments and stomatal conductance was more profound in SARC 5 than in SARC 1. Combined application of 10 mM K and HA was most effective in alleviating salt stress with improved plant biomass by 47% and 43% in SARC 1 and SARC 5, respectively. Combined application of 10 mM K and HA mitigated salt and induced oxidative stress with the activities of APX, CAT, POD and SOD increased by up to 2.8 folds in SARC 1, and by upto 2.5 folds in SARC 5, respectively. Root and shoot Na contents were increased, while K, Fe and Zn contents were decreased under saline conditions. HA combined with K decreased Na and increased K, Fe and Zn contents in both genotypes. Combined application of 10 mM K and HA was more promising for increasing wheat salt tolerance and nutrient uptake and genotype SARC 1 performed better than SARC 5 for cultivation on saline soils.

Heat stress during reproductive and grain filling phases adversely affects the growth of cereals through reduction in grain’s number and size. However, exogenous application of antioxidants, plant growth regulators and osmoprotectants may be helpful to minimize these heat induced yield losses in cereals. This two year study was conducted to evaluate the role of exogenous application of ascorbic acid (AsA), salicylic acid (SA) and hydrogen peroxide (H2O2) applied through seed priming or foliar spray on biochemical, physiological, morphological and yield related traits, grain yield and quality of late spring sown hybrid maize. The experiment was conducted in the spring season of 2007 and 2008. We observed that application of AsA, SA and H2O2 applied through seed priming or foliar spray improved the physiological, biochemical, morphological and yield related traits, grain yield and grain quality of late spring sown maize in both years. In both years, we observed higher superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activity in the plants where AsA, SA and H2O2were applied through seed priming or foliar spray than control. Membrane stability index (MSI), relative water contents (RWC), chlorophyll contents, grain yield and grain oil contents were also improved by exogenous application of AsA, SA and H2O2 in both years. Seed priming of AsA, SA and H2O2was equally effective as the foliar application. In conclusion, seed priming with AsA, SA and H2O2 may be opted to lessen the heat induced yield losses in late sown spring hybrid maize. Heat tolerance induced by ASA, SA and H2O2 may be attributed to increase in antioxidant activities and MSI which maintained RWC and chlorophyll contents in maize resulting in better grain yield in heat stress conditions.

Abstract Background Jalili syndrome (JS) is a rare cone‐rod dystrophy (CRD) associated with amelogenesis imperfecta (AI). The first clinical presentation of JS patients was published in 1988 by Jalili and Smith. Pathogenic mutations in the Cyclin and CBS Domain Divalent Metal Cation Transport Mediator 4 (CNNM4) magnesium transporter protein have been reported as the leading cause of this anomaly. Methods In the present study, a clinical and genetic investigation was performed in a consanguineous family of Pakistani origin, showing characteristic features of JS. Sanger sequencing was successfully used to identify the causative variant in CNNM4. Molecular dynamics (MD) simulations were performed to study the effect of amino acid change over CNNM4 protein. Results Sequence analysis of CNNM4 revealed a novel missense variant (c.1220G>T, p.Arg407Leu) in exon‐1 encoding cystathionine‐β‐synthase (CBS) domain. To comprehend the mutational consequences in the structure, the mutant p.Arg407Leu was modeled together with a previously reported variant (c.1484C>T, p.Thr495Ile) in the same domain. Additionally, docking analysis deciphered the binding mode of the adenosine triphosphate (ATP) cofactor. Furthermore, 60ns MD simulations were carried out on wild type (p.Arg407/p.Thr495) and mutants (p.Arg407Leu/p.Thr495Ile) to understand the structural and energetic changes in protein structure and its dynamic behavior. An evident conformational shift of ATP in the binding site was observed in simulated mutants disrupting the native ATP‐binding mode. Conclusion The novel identified variant in CNNM4 is the first report from the Pakistani population. Overall, the study is valuable and may give a novel insight into metal transport in visual function and biomineralization.

Abstract Background Hearing loss or hearing impairment is a clinically and genetically heterogeneous disorder. More than 117 genes were discovered to date in hereditary, nonsyndromic hearing loss (NSHL). Identifying novel gene variants and their frequency in specific populations is valuable for public health and potentially for genetic screening of NSHL. Aims To identify the gene variants underlying NSHL in a Pakistani cohort. Methods and Results A cohort of 40 school‐aged children with NSHL was initially screened for variants in GJB2, the gene with the highest incidence of variants in other populations with NSHL. We found known homozygous as well as compound heterozygous GJB variants in 15 individuals. Next, we used targeted next generation sequencing (TNGS) for the remaining 25 individuals and identified 20 different variants in 14 genes (SLC26A4, KCNQ4, MYO7A, MYO15A, TMPRSS3, ESPN, TMC1, GIPC3, LHFPL5, WFS1, DFNB59, GRXCR1, ESRRB, and LRTOMT). Conclusions We described common and novel variants in 15 genes in a Pakistani cohort of NSHL.

Abstract Planting arrangements affect radiation use efficiency (RUE) and competitiveness of intercrop species in intercropping systems. Here, we reveal that narrow‐wide‐row planting arrangement in maize‐soybean relay‐intercropping system increases the dry matter and competitiveness of soybean, increased the RUE of maize and soybean, and compensates the yield loss of maize by substantially increasing the yield of soybean. In this field study, maize was planted with soybean in different planting arrangements (P1, 20:180, P2, 40:160; P3, 60:140, and P4, 80:120) of relay intercropping, all the relay‐intercropping treatments were compared with sole crops of maize (SM) and soybean (SS). Results showed that P1 improved the total RUE 3.26 g/MJ (maize RUE + soybean RUE) of maize and soybean in relay‐intercropping system. Compared to P4, treatment P1 increased the soybean competition ratio (CR) values (by 55%) but reduced the maize CR values (by 29%), which in turn significantly improved the yield of soybean by maintaining the maize yield. Generally, in P1, soybean produced 82% of SS yield, and maize produced 88% of SM yield, and it achieved the land equivalent ratio of 1.7. These results suggest that by maintaining the appropriate planting distances between maize and soybean we can improve the competitiveness and yield of intercrop species in relay‐intercropping system.

Background Maize-soybean relay-intercropping (MSR) is a famous system of crop production in developing countries. However, maize shading under this system directly affects the light quality and intensity of soybean canopy. This is a challenging scenario in which to implement the MSR system, in terms of varieties selection, planting pattern, and crop management since the duration of crop resource utilization clearly differs. Methods Therefore, this experiment aimed to elucidate the effect of leaf excising treatments from maize top to fully clarify the needs and balance of light quality and intensity of intercrop-soybean under MSR in field conditions. The effects of different leaf excising treatments (T0, no removal of leaves; T2, removal of two topmost leaves; T4, removal of four topmost leaves; T6, removal of six topmost leaves from maize plants were applied at first-trifoliate stage (V1) of soybean) on photosynthetically active radiation transmittance (PART), red to far-red ratio (R:FR), morphological and photosynthetic characteristics and total biomass production at second-trifoliate stage (V2), fifth-trifoliate stage (V5), and flowering-stage (R1) of soybean were investigated through field experiments for 2-years under MSR. Results As compared to T0, treatment T6 increased the PART and R:FR ratio at soybean canopy by 77% and 37% (V2), 70% and 34% (V5), and 41% and 36% (R1), respectively. This improved light environment in T6 considerably enhanced the leaf area index, SPAD values and photosynthetic rate of soybean plants by 66%, 25% and 49% at R1, respectively than T0. Similarly, relative to control, T6 also increased the stem diameter (by 29%) but decreased the plant height (by 23%) which in turn significantly increased stem breaking strength (by 87%) by reducing the lodging rate (by 59%) of soybean plants. Overall, under T6, relay-cropped soybean produced 78% of sole soybean seed-yield, and relay-cropped maize produced 81% of sole maize seed-yield. Our findings implied that by maintaining the optimum level of PART (from 60% to 80%) and R:FR ratio (0.9 to 1.1), we can improve morphological and photosynthetic characteristics of soybean plants in MSR. Therefore, more attention should be paid to the light environment when considering the sustainability of MSR via appropriate planting pattern selection.

Salinity is a major abiotic stress which limits crop production, especially under rainfed conditions. Selenium (Se), as an important micronutrient, plays a vital role in mitigating detrimental effects of different abiotic stresses. The objective of this research was to examine the effect of Se fertilization on black gram (Vigna mungo) under salt stress. Our results showed that salt stress (100 mM NaCl) in leaves significantly induced oxidative damage and caused a decline in relative water content, chlorophyll (Chl), stomatal conductance (gs), photochemical efficiency (Fv/Fm), sucrose, and reducing sugars. A low dose of Se (1.5 ppm) significantly reduced hydrogen peroxide content, malondialdehyde formation, cell membrane damage, and also improved antioxidative enzyme activities, including superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and glutathione peroxidase under salt stress. Se-treated plants exhibited higher Chl, gs, Fv/Fm, sucrose, and reducing sugars than untreated plants in response to salt stress. In addition, Se application enhanced Se uptake and reduced Na+ uptake, but Cl− remained unaffected. Our results indicated that a low dose of Se effectively alleviated salt damage via inhibition of Na+ uptake and enhanced antioxidant defense resulting in a significant decrease in oxidative damage, and maintained gaseous exchange and PS II function for sucrose and reducing sugars accumulation in black gram.

A better understanding of plant mechanisms in response to drought is a strong premise to achieving high yields while saving unnecessary water. This is especially true in the case of biomass crops for non-food uses (energy, fibre and forage), grown with limited water supply. In this frame, we investigated growth and physiological response of two genotypes of biomass sorghum (Sorghum bicolor (L.) Moench) to contrasting levels of soil moisture in a pot experiment carried out in a greenhouse. Two water regimes (high and low water, corresponding to 70% and 30% field capacity) were applied to JS-2002 and Trudan-8 sorghum genotypes, respectively bred for dry sub-tropical and mild temperate conditions. Two harvests were carried out at 73 and 105 days after seeding. Physiological traits (transpiration, photosynthesis and stomatal conductance) were assessed in four dates during growth. Leaf water potential, its components and relative water content were determined at the two harvests. Low watering curbed plant height and aboveground biomass to a similar extent (ca. 􀀀70%) in both genotypes. JS-2002 exhibited a higher proportion of belowground to aboveground biomass, i.e., a morphology better suited to withstand drought. Despite this, JS-2002 was more affected by low water in terms of physiology: during the growing season, the average ratio in transpiration, photosynthesis and stomatal conductance between droughty and well watered plants was, respectively, 0.82, 0.80 and 0.79 in JS-2002; 1.05, 1.08 and 1.03 in Trudan-8. Hence Trudan-8 evidenced a ca. 20% advantage in the three traits. In addition, Trudan-8 could better exploit abundant moisture (70% field capacity), increasing aboveground biomass and water use efficiency. In both genotypes, drought led to very low levels of leaf water potential and relative water content, still supporting photosynthesis. Hence, both morphological and physiological characteristics of sorghum were involved in plant adaptation to drought, in accordance with previous results. Conversely, the common assumption that genotypes best performing under wet conditions are less suited to face drought was contradicted by the results of the two genotypes in our experiment. This discloses a potential to be further exploited in programmes of biomass utilization for various end uses, although further evidence at greenhouse and field level is needed to corroborate this finding.

Oilseeds yield response to sulphur (S) has been well investigated but the dynamics of total biomass accumulation (TBA) and partitioning by sesame plants in response to S are not well understood. This study was initiated to investigate the effects of S on sesame, in which four sesame varieties V1, SG-27; V2, SG-30; V3, SG-36; V4, SG-51 and four S treatments T1, 20; T2, 30; T3, 40; T4, 50 kg ha−1 were used. Results revealed that the leaf area index and photosynthetic rate of sesame varieties were significantly higher under T3 with V3. Similarly, S fertilization considerably increased the TBA and maximum TBA was reached at late-flowering with V3 in T3. Relative to T1, plants in T3 had 33 and 23% higher capsule and seed biomass, respectively. Furthermore, total S accumulation and distribution in different plant organs changed with growth stage, at pre-flowering and mid-flowering stage maximum S was found in the stem, whereas at late-flowering and full-maturity stage highest S was recorded in reproductive parts. These results implied that S fertilizers should be applied to agricultural fields to improve oilseed production and by selecting the appropriate and area-specific genotype we can increase sesame seed yield under rainfed conditions.

Silicon (Si) serves as bioactive beneficial element. Si is highly abundant in soil, and occurs ubiquitously in all organisms including plants and humans. During the last three decades, nutritional significance of Si for plant and human health has received increasing attention. Plant Si plays a pivotal role in growth and development, and this beneficial effect depends usually on accumulation in plant tissues, which are then protected from various forms of biotic and abiotic stresses. Likewise, human exposure to Si imparts health benefits and essentially occurs through plant-derived food products. Si bioavailability in human diet e.g. strengthens bones and improves immune response, as well as neuronal and connective tissue health. Despite this empiric knowledge, the essentiality of Si still remains enigmatic. Thus the link between Si availability for plant development and its profound implication for human welfare should receive attention. This review aims to provide a broad perspective on Si as important element for plant and human nutrition and to define research fields for interdisciplinary research.

Ectodermal dysplasias (EDs) are a large heterogeneous group of inherited disorders exhibiting abnormalities in ectodermally derived appendages such as hair, nails, teeth and sweat glands. EDs associated with reticulated pigmentation phenotype are rare entities for which the genetic basis and pathophysiology are not well characterized. The present study describes a five generation consanguineous Pakistani family segregating an autosomal recessive form of a novel type of ectodermal dysplasia. The affected members present with sparse and woolly hair, severe nail dystrophy and reticulate skin pigmentation. After exclusion of known gene loci related with other skin disorders, genome-wide linkage analysis was performed using Illumina HumanOmniExpress beadchip SNP arrays. We linked this form of ED to human chromosome 18p11.32-p11.31 flanked by the SNPs rs9284390 (0.113Mb) and rs4797100 (3.14 Mb). A maximum two-point LOD score of 3.3 was obtained with several markers along the disease interval. The linkage interval of 3.03 Mb encompassed seventeen functional genes. However, sequence analysis of all these genes did not discover any potentially disease causing-variants. The identification of this novel locus provides additional information regarding the mapping of a rare form of ED. Further research, such as the use of whole-genome sequencing, would be expected to reveal any pathogenic mutation within the disease locus.