Homozygous CCDC146 mutation causes oligoasthenoteratozoospermia in humans and mice

Infertility represents a significant health concern,with sperm quantity and quality being crucial determinants of male fertility.Oligoasthenoteratozoospermia(OAT)is characterized by reduced sperm motility,lower sperm concentration,and morphological abnormalities in sperm heads and flagella.Although variants in several genes have been implicated in OAT,its genetic etiologies and pathogenetic mechanisms remain inadequately understood.In this study,we identified a homozygous nonsense mutation(c.916C>T,p.Arg306*)in the coiled-coil domain containing 146(CCDC146)gene in an infertile male patient with OAT.This mutation resulted in the production of a truncated CCDC146 protein(amino acids 1-305),retaining only two out of five coiled-coil domains.To validate the pathogenicity of the CCDC146 mutation,we generated a mouse model(Ccdc146^(mut/mut))with a similar mutation to that of the patient.Consistently,the Ccdc146mut/mut mice exhibited infertility,characterized by significantly reduced sperm counts,diminished motility,and multiple defects in sperm heads and flagella.Furthermore,the levels of axonemal proteins,including DNAH17,DNAH1,and SPAG6,were significantly reduced in the sperm of Ccdc146^(mut/mut) mice.Additionally,both human and mouse CCDC146 interacted with intraflagellar transport protein 20(IFT20),but this interaction was lost in the mutated versions,leading to the degradation of IFT20.This study identified a novel deleterious homozygous nonsense mutation in CCDC146 that causes male infertility,potentially by disrupting axonemal protein transportation.These findings offer valuable insights for genetic counseling and understanding the mechanisms underlying CCDC146 mutant-associated infertility in human males.

Foliar application of micronutrients enhances crop stand, yield and the biofortification essential for human health of different wheat cultivars

Globally about half of the world’s population is under micronutrient malnutrition due to poor quality food intake.To overcome this problem,fortification and biofortification techniques are often used.Biofortification is considered a better option than fortification due to the easy control of nutrient deficiencies present in daily food.This field experiment was conducted to evaluate the effects of foliar application of a micronutrient mixture(MNM)consisting of zinc(Zn),iron(Fe),copper(Cu),manganese(Mn)and boron(B)on yield and flour quality of wheat.The results show the effectiveness of foliar feeding for growth and yield parameters,in addition to the enriching of wheat grains with Zn,Cu,Fe,Mn and B.Compared to the control without foliar feeding,foliar application on wheat crop increased tillering ability,spike length,grain yield and the contents of Zn,Cu,Mn,Fe and B by 21,47,22,22 and 25%in wheat flour,respectively.Therefore,foliar feeding of micronutrients could be an effective approach to enrich wheat grains with essential nutrients for correcting malnutrition.

Uncertainty analysis of runoff and sedimentation in a forested watershed using sequential uncertainty fitting method

The Soil and Water Assessment Tool （SWAT） was implemented in a small forested watershed of the Soan River Basin innorthern Pakistan through application of the sequential uncertainty fitting （SUFI-2） method to investigate the associateduncertainty in runoff and sediment load estimation. The model was calibrated for a 10-year period （1991–2000） with aninitial 4-year warm-up period （1987–1990）, and was validated for the subsequent 10-year period （2001–2010）. Themodel evaluation indices R2 （the coefficient of determination）, NS （the Nash-Sutcliffe efficiency）, and PBIAS （percentbias） for stream flows simulation indicated that there was a good agreement between the measured and simulated flows.To assess the uncertainty in the model outputs, p-factor （a 95% prediction uncertainty, 95PPU） and r-factors （averagewideness width of the 95PPU band divided by the standard deviation of the observed values） were taken into account.The 95PPU band bracketed 72% of the observed data during the calibration and 67% during the validation. The r-factorwas 0.81 during the calibration and 0.68 during the validation. For monthly sediment yield, the model evaluation coefficients（R2 and NS） for the calibration were computed as 0.81 and 0.79, respectively; for validation, they were 0.78and 0.74, respectively. Meanwhile, the 95PPU covered more than 60% of the observed sediment data during calibrationand validation. Moreover, improved model prediction and parameter estimation were observed with the increasednumber of iterations. However, the model performance became worse after the fourth iterations due to an unreasonableparameter estimation. Overall results indicated the applicability of the SWAT model with moderate levels of uncertaintyduring the calibration and high levels during the validation. Thus, this calibrated SWAT model can be used for assessmentof water balance components, climate change studies, and land use management practices.

Computationally predicted pathogenic USP9X mutation identified in infertile men does not affect spermatogenesis in mice

Infertility is a major health issue,affecting approximately 15%of couples of child-bearing age.Although nearly half of idiopathic infertility cases are assumed to have a genetic basis,the underlying causes remain largely unknown in most infertile men.

Comparative study of probable maximum precipitation and isohyetal maps for mountainous regions, Pakistan

Probable maximum precipitation(PMP) is widely used by hydrologists for appraisal of probable maximum flood(PMF)used for soil and water conservation structures, and design of dam spillways. A number of methods such as empirical, statistical and dynamic are used to estimate PMP, the most favored being statistical and hydro-meteorological. In this paper,PMP estimation in mountainous regions of Pakistan is studied using statistical as well as physically based hydro-meteorological approaches. Daily precipitation, dew point, wind speed and temperature data is processed to estimate PMP for a one-day duration. Maximum precipitation for different return periods is estimated by using statistical approaches such as Gumble and Log-Pearson type-III(LP-III) distribution. Goodness of fit(GOF) test, chi-square test, correlation coefficient and coefficient of determination were applied to Gumble and LP-III distributions. Results reveal that among statistical approaches, Gumble distribution performed the best result compared to LP-III distribution. Isohyetal maps of the study area at different return periods are produced by using the GIS tool, and PMP in mountainous regions varies from 150 to 320 mm at an average value of 230.83 mm. The ratio of PMP for one-day duration to highest observed rainfall(HOR) varied from 1.08 to 1.29 with an average value of 1.18. An appropriate frequency factor(K_m) is very important which is a function of mean for observed precipitation and PMP for 1-day duration, and K_m values varies from 2.54 to 4.68. The coefficient of variability(C_v) varies from minimum value of 28% to maximum value of 43.35%. It was concluded that the statistical approach gives higher results compared to moisture maximization(MM) approach. In the hydro-meteorological approach, moisture maximization(MM) and wind moisture maximization(WMM) techniques were applied and it was concluded that wind moisture maximization approach gives higher results of PMP as compared to moisture maximization approach as well as for Hershfield technique. Therefore, it is suggested that MM approach is the most favored in the study area for PMP estimation, which leads to acceptable results, compared to WMM and statistical approaches.

Bioceutical Properties of Culinary Herbs, Spices and Their Possible Outcomes with Standard Antibiotics

Culinary condiments have been used for centuries to treat several types of ailments. Four ethanolic lipid fractions including Nigella sativa, Foeniculum vulgare, Laurus nobilis, and Coriandrum sativum were selected to assess their antimicrobial potential alone and combined with antibiotics. Antibacterial activity was determined by various conventional procedures such as aromatogram test, well diffusion, macro-broth dilution, disc diffusion assay. The tested lipid fractions exhibited antibacterial activity against selected bacterial strains. Antibacterial activity of lipid fractions was also seen within two hours of incubation. These lipid fractions attacked the cell wall i.e. the penetrability of bacterial cell and hence intracellular contents released in the environment which was detected at 260 nm of absorbance and verified via scanning electron microscopy. Antagonistic effect was mostly found by the combination of antibiotics and lipid fractions. Though, synergistic effect was obtained with beta-lactam drugs when combined with lipid fraction of Laurus nobilis. Hence the lipid fractions may be used to treat various communicable diseases. However antagonistic outcomes of the combinations suggested that herbal and allopathic therapies should not consume concurrently.

Investigation of Iron Complex Formation of Anti-Hypertensive Drug: Methyldopa

The drug administered for any disease may play an unwanted function in biological system. They may have multiple counter effects, one of which is their interaction to bioactive metals. Iron is most common bio essential metal and is reported to interact with antihypertensive drug methyldopa. In the present study, above said complex is analyzed by UV-Visible spectrophotometry. Formation constant of the complex is calculated by using mole ratio method and single point statistical method which is in the range of 1010, values are also calculated which are independent of pH like formation constant. Absorbance maxima were found to be dependent on pH. At lower pH complex shows two broad bands centered at 430 nm and 730 nm. With the rise in pH later peak shifts toward lower wavelength, so 615 nm is selected for further studies. Molar extinction coefficient of the complex is explored by serial dilution method. At all wavelengths it increases with increase in pH. Mole ratio and slope ratio methods are used for exploring stoichiometry. Metal to ligand combining ratio in the complex is 1:2 at pH 4.0 and pH 4.5 while 1:3 at pH 5.0 and pH 5.5.

A novel missense mutation of CCDC34 causes male infertility with oligoasthenoteratozoospermia in a consanguineous Pakistani family

Male infertility is a worldwide health issue,affecting 8%–12%of the global population.Oligoasthenoteratozoospermia(OAT)represents a severe type of male infertility,characterized by reduced sperm count and motility and an increased frequency of sperm with aberrant morphology.Using whole-exome sequencing,this study identified a novel missense mutation(c.848C>A,p.A283E)in the coiled-coil domain-containing 34 gene(CCDC34)in a consanguineous Pakistani family.This rare mutation was predicted to be deleterious and to affect the protein stability.Hematoxylin and eosin staining of spermatozoa from the patient with OAT revealed multiple morphological abnormalities of the flagella and transmission electron microscopy indicated axonemal ultrastructural defects with a lack of outer dynein arms.These findings indicated that CCDC34 plays a role in maintaining the axonemal ultrastructure and the assembly or stability of the outer dynein arms,thus expanding the phenotypic spectrum of CCDC34 missense mutations.

A recurrent homozygous missense mutation in CCDC103 causes asthenoteratozoospermia due to disorganized dynein arms

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

Parameter evaluation for soil erosion estimation on small watersheds using SWAT model

This research was undertaken for the evaluation of soil erosion using the semi-distributed basin scale SWAT model for four subcatchments of the Dhrabi River Catchment(DRC),which is located in the Pothwar Plateau region.Two subcatchments(catchment-25 and-31)are characterized by gullies while the other two(catchment-27 and-32)are managed with terraced landuse system.The performance of the model was satisfactory with coefficient of determination(R^(2))=0.67 to 0.91 and Nash-Sutcliffe efficiency(ENS)=0.54 to 0.85 for both surface runoff and sediment yield during the calibration(2009-2010)and validation(2011)periods.The PUSLE factor was found to be the most sensitive parameter during model calibration.It was observed that all of the rainfall-runoff events occurred during the monsoon season(June to September).The estimated annual sediment loss ranged from 2.6 t/hm^(2) to 31.1 t/hm^(2) over the duration of the simulation period for the non-terraced catchments,in response to annual precipitation amounts that were between 194.8 mm to 579.3 mm.In contrast,the predicted annual sediment levels for the terraced catchments ranged from 0.52 t/hm^(2) to 10.10 t/hm^(2) due to similar precipitation amounts.The terraced catchments resulted in 4 to 5 times lower sediment yield as compared to non-terraced catchments.The results suggest that there is a huge potential for terraces to reduce soil erosion in the DRC specifically and Pothwar area generally,which have proven to be an efficient approach to establishing soil and water conservation structures in this region.