Multiple morphological abnormalities of the flagella (MMAF) represent a severe form of sperm defects leading to asthenozoospermia and male infertility. In this study, we identified a novel homozygous splicing mutation (c.871-4 ACA>A) in the adenylate kinase 7 (AK7) gene by whole-exome sequencing in infertile individuals. Spermatozoa from affected individuals exhibited typical MMAF characteristics, including coiled, bent, short, absent, and irregular flagella. Transmission electron microscopy analysis showed disorganized axonemal structure and abnormal mitochondrial sheets in sperm flagella. Immunofluorescence staining confirmed the absence of AK7 protein from the patients' spermatozoa, validating the pathogenic nature of the mutation. This study provides direct evidence linking the AK7 gene to MMAF-associated asthenozoospermia in humans, expanding the mutational spectrum of AK7 and enhancing our understanding of the genetic basis of male infertility.
Humans ; Male ; Sperm Tail/ultrastructure* ; Homozygote ; Consanguinity ; Asthenozoospermia/pathology* ; Infertility, Male/genetics* ; Mutation ; Pakistan ; Adenylate Kinase/genetics* ; Adult ; Pedigree ; RNA Splicing ; Exome Sequencing ; Spermatozoa

Male infertility can result from impaired sperm motility caused by multiple morphological abnormalities of the flagella (MMAF). Distinct projections encircling the central microtubules of the spermatozoal axoneme play pivotal roles in flagellar bending and spermatozoal movement. Mammalian sperm-associated antigen 17 ( SPAG17 ) encodes a conserved axonemal protein of cilia and flagella, forming part of the C1a projection of the central apparatus, with functions related to ciliary/flagellar motility, skeletal growth, and male fertility. This study investigated two novel homozygous SPAG17 mutations (M1: NM_206996.2, c.829+1G>T, p.Asp212_Glu276del; and M2: c.2120del, p.Leu707*) identified in four infertile patients from two consanguineous Pakistani families. These patients displayed the MMAF phenotype confirmed by Papanicolaou staining and scanning electron microscopy assays of spermatozoa. Quantitative real-time polymerase chain reaction (PCR) of patients' spermatozoa also revealed a significant decrease in SPAG17 mRNA expression, and immunofluorescence staining showed the absence of SPAG17 protein signals along the flagella. However, no apparent ciliary-related symptoms or skeletal malformations were observed in the chest X-rays of any of the patients. Transmission electron microscopy of axoneme cross-sections from the patients showed incomplete C1a projection and a higher frequency of missing microtubule doublets 1 and 9 compared with those from fertile controls. Immunofluorescence staining and Western blot analyses of spermatogenesis-associated protein 17 (SPATA17), a component of the C1a projection, and sperm-associated antigen 6 (SPAG6), a marker of the spring layer, revealed disrupted expression of both proteins in the patients' spermatozoa. Altogether, these findings demonstrated that SPAG17 maintains the integrity of spermatozoal flagellar axoneme, expanding the phenotypic spectrum of SPAG17 mutations in humans.
Humans ; Male ; Infertility, Male/pathology* ; Sperm Tail/ultrastructure* ; Homozygote ; Microtubule-Associated Proteins/genetics* ; Axoneme/genetics* ; Spermatozoa/ultrastructure* ; Adult ; Mutation ; Sperm Motility/genetics* ; Pedigree ; Microtubules ; Microtubule Proteins/genetics*

Multiple morphological abnormalities of sperm flagella (MMAF) is a severe form of asthenoteratozoospermia, characterized by morphological abnormalities and reduced motility of sperm, causing male infertility. Although approximately 60% of MMAF cases can be explained genetically, the etiology of the remaining cases is unclear. Here, we identified two novel compound heterozygous variants in the gene, dynein axonemal heavy chain 10 ( DNAH10 ), in three patients from two unrelated Pakistani families using whole-exome sequencing (WES), including one compound heterozygous mutation ( DNAH10 : c.9409C>A [p.P3137T]; c.12946G>C [p.D4316H]) in family 1 and another compound heterozygous mutation ( DNAH10 : c.8849G>A [p.G2950D]; c.11509C>T [p.R3687W]) in family 2. All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools. Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance. Hematoxylin and eosin (H&E) staining revealed MMAF, including sperm head abnormalities, in the patients. In addition, immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella. These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.
Adult ; Humans ; Male ; Alleles ; Asthenozoospermia/pathology* ; Axonemal Dyneins/genetics* ; Dyneins/genetics* ; Exome Sequencing ; Infertility, Male/pathology* ; Mutation ; Pakistan ; Pedigree ; Sperm Tail/pathology*

The syndrome of multiple morphological abnormalities of the sperm flagella (MMAF) is one of the most serious kinds of sperm defects, leading to asthenoteratozoospermia and male infertility. In this study, we use whole-exome sequencing to identify genetic factors that account for male infertility in a patient born from a consanguineous Pakistani couple. A homozygous frameshift mutation (c.1399_1402del; p.Gln468ArgfsTer2) in axonemal dynein light chain domain containing 1 ( AXDND1 ) was identified in the patient. Sanger sequencing data showed that the mutation was cosegregated recessively with male infertility in this family. Papanicolaou staining and scanning electron microscopy analysis of the sperm revealed severely abnormal flagellar morphology in the patient. Immunofluorescence and western blot showed undetectable AXDND1 expression in the sperm of the patient. Transmission electron microscopy analysis showed disorganized sperm axonemal structure in the patient, particularly missing the central pair of microtubules. Immunofluorescence staining showed the absence of sperm-associated antigen 6 (SPAG6) and dynein axonemal light intermediate chain 1 (DNALI1) signals in the sperm flagella of the patient. These findings indicate that AXDND1 is essential for the organization of flagellar axoneme and provide direct evidence that AXDND1 is a MMAF gene in humans, thus expanding the phenotypic spectrum of AXDND1 frameshift mutations.
Humans ; Male ; Sperm Tail/ultrastructure* ; Frameshift Mutation ; Infertility, Male/pathology* ; Pakistan ; Pedigree ; Consanguinity ; Axonemal Dyneins/genetics* ; Adult ; Spermatozoa ; Exome Sequencing

Objectives: Postmenopausal females often experience genitourinary symptoms like vulvovaginal dryness due to estrogen decline.Hormone replacement therapy is effective in alleviating vaginal atrophy and genitourinary syndrome in this population. Evaluate local estrogen’s safety and effectiveness for alleviating postmenopausal vaginal symptoms, including endometrial thickness, dyspareunia, vaginal pH, and dryness. Methods: We searched Google Scholar, Cochrane Library, ClinicalTrial.Gov, PubMed, and ScienceDirect databases until July 2023. All randomized controlled trials (RCTs) linking intravaginal estrogen supplementation to vaginal atrophy or vaginitis were included. The risk of bias was evaluated with RoB 2, and publication bias was assessed using Egger and Beggs analysis. Results: All evidence pertains to females. Eighteen studies (n = 4,723) compared estrogen with placebo. Patients using estrogen showed a significant increase in superficial cells (mean differences [MD]: 19.28; 95% confidence intervals [CI]: 13.40 to 25.16; I2 = 90%; P < 0.00001) and a decrease in parabasal cells (MD: –24.85; 95% CI: –32.96 to –16.73; I2 = 92%; P < 0.00001). Vaginal pH and dyspareunia significantly reduced in estrogen users (MD: –0.94; 95% CI: –1.05 to –0.84; I2 = 96%) and (MD: –0.52; 95% CI: –0.63 to –0.41; I2 = 99%), respectively. Estrogen did not significantly affect vaginal dryness (MD: –0.04; 95% CI: –0.18 to 0.11; I2 = 88%). Adverse events like vulvovaginal pruritis, mycotic infection, and urinary tract infection were reported, but the association was insignificant (risk ratio: 0.95; 95% CI: 0.88 to 1.02; I2 = 0%). Conclusions Our meta-analysis of 18 RCTs suggests promising potential for intravaginal estrogen therapy in alleviating vaginal atrophy and vaginitis in postmenopausal females.

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenozoospermia categorized by immotile spermatozoa with abnormal flagella in ejaculate. Whole-exome sequencing (WES) is used to detect pathogenic variants in patients with MMAF. In this study, a novel homozygous frameshift variant (c.6158_6159insT) in dynein axonemal heavy chain 8 (DNAH8) from two infertile brothers with MMAF in a consanguineous Pakistani family was identified by WES. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed DNAH8 mRNA decay in these patients with the DNAH8 mutation. Hematoxylin-eosin staining and transmission electron microscopy revealed highly divergent morphology and ultrastructure of sperm flagella in these patients. Furthermore, an immunofluorescence assay showed the absence of DNAH8 and a reduction in its associated protein DNAH17 in the patients' spermatozoa. Collectively, our study expands the phenotypic spectrum of patients with DNAH8-related MMAF worldwide.
Humans ; Male ; Consanguinity ; Pakistan ; Infertility, Male/metabolism* ; Semen/metabolism* ; Sperm Tail/metabolism* ; Spermatozoa/metabolism* ; Flagella/pathology* ; Mutation

Objective: To test the relative efficacy of pyriproxyfen and methoprene on mortality, deformity, inhibition and emergence to adult stages of Culex quinquefasciatus and Aedes albopictus. Methods: Serial dilutions (0.01-0.05 mg/L) of methoprene, pyriproxyfen 0.5 water dispersible granules (WDG) and pyriproxyfen 1.0 WDG were used to assess mortality and inhibition of 3rd instar larvae of Aedes albopictus and Culex quinquefasciatus. Each concentration and control was replicated four times in completely randomized design. Data on larval mortality, growth inhibition, deformities and adult's emergence was recorded weekly. On the basis of best comparative performance, the efficacy of pyriproxyfen 1.0 WDG at 0.1 g/m

Objective: To understand the impact of platelet associated immunoglobulin G (PAIgG)/platelet associated immunoglobulin M (PAIgM) on severity of dengue virus infection leading to thrombocytopenia. Methods: In this study we examined a total of 52 patients who were having secondary infection of dengue in acute phase by using competitive ELISA. Results: A decrease in the platelet count was observed at the acute phase of infection while all along the recovery stage the count of platelet was significantly increased. A significant decrease was observed in PAIgG and PAIgM in these subjects. Inverse correlation was found between platelets count and PAIgG/PAIgM among the subjects studied. In the platelets elution from ten subjects, anti-dengue virus immunoglobulin G and immunoglobulin M were observed. PAIgG and PAIgM with inclined levels were higher in dengue hemorrhagic fever than the classical dengue fever. In the development of dengue hemorrhagic fever PAIgM inclined level was independently associated with high specificity, showing a possible indication of dengue hemorrhagic fever. Conclusions: This study suggests that in secondary dengue virus infection, the PAIgG and PAIgM levels, and the activity of anti-dengue virus play key roles, both in the development and severity of the disease.

AIM: To study the pattern of presentation and to highlight the common causes of primary benign orbital lesions.of Jinnah Postgraduate Medical Centre Karachi from July 1997 to August 2001 and then from September 2001 to date (Continued) at the Department of Ophthalmology of Chandka Medical College & Hospital Larkana. Only patients with primary benign orbital lesions were included in this study. All the patients were admitted in eye ward from the out patients department. The diagnosis of the disease was based on the presentation, clinical examination, investigations and histopathology of excised mass. A total of 68 patients were included in this study. The age range was from 2 months to 60 years. Out of 68, 27 (39.7%) patients were male and 41 (60.3%) were female. The left orbit was involved in 35 (51.5%) and right orbit was involved in 33 (48.5%).revealed that superficial capillary hemangiomas of the eye lid were the most common lesion 26 (38.2%) followed by deep orbital cavernous hemangiomas 5 (7.4%), lymphangiomas 5 (7.4%), orbital varices 4 (5.9%), gliomas 7 (10.3%), meningiomas 5 (7.4%), neurofibromas 5 (7.4%), neurofibromatosis 4 (5.9%), schwannomas 2 (2.9%), and pleomorphic adenoma (benign mixed cell tumor) of lacrimal gland 5 (7.4%) cases.treatment can prevent the patient from visual and life threatening complications.

AIM:To evaluate the site, size,histology,and recurrence of the tumor.METHODS: We prospectively evaluated 53 patients with histological diagnosis of retinoblastoma involving the eye, orbit, intra cranial cavity and other tissues, at Department of Ophthalmology, Chandka Medical College Hospital, Larkana. To assess the extent of the tumor investigations, including X-ray-orbits, ocular B-scan, CT-scan/MRI-Oorbits-brain, whole body bone scanning, X-ray chest PA view, ultrasound abdomen, lumbar punc-ture, complete blood cell count, blood chemistry as-sessment, including renal and hepatic function para-meters, were performed. The selection of mode of treatment like surgical procedures, local radiotherapy and systemic chemotherapy used depend on the site, size, recurrence and metastasis of the tumor. The patients with local and systemic metastases were referred to the oncologist at the Larkana Institute of the nuclear medicine and radiotherapy for local radiation and systemic chemotherapy. Follow up examinations were carried out at 1, 3 , 6 and 12 months after the operation, and then annually for a further 5 years or longer. The tumor's site, size, histological type, and recurrence were evaluated.RESULTS: Of these 53 patients the age range was from 8 months to 8 years. 25 (47%) were male and 28 (53%) were female. The laterality of the tumor was in left eye 25 (47%) cases, right eye 19 (36%) cases, and both eyes 9 (17%) cases. The presentation of the patient at 1st visit was strabismus 3 (6%) cases, pseudohypopyon 4 (8%) cases, orbital pseudo orbital inflammation 6 cases (11%), mass with metastases 7 (13%) cases, fungating mass 8 (15%) cases, leu-kocoria 12(23%) cases and orbital invasion 13(25%) cases. The surgical procedures performed were enucleation alone in 19 (36%) cases, enucleation with modified exentration followed by local radiotherapy and systemic chemotherapy in 19 (36%) cases and enucleation with total exentration followed by local radiotherapy and systemic chemotherapy in 15 (28%) cases. The histological types of the tumor were well- differentiated retinoblastoma with abundant rosettes in 25 (47%) cases and highly undifferentiated retino- blastoma 28 (53%) cases . After primary surgery, the recu-rrences and metastases of the tumors were noticed in 37 (70%) cases due to optic nerve involvement beyond the point of surgical transaction, orbital invasion and sy-stemic metastasis.CONCLUSION: Any child with red eye or white pupillary reflex should be suspected of having retinoblastoma, until proved otherwise, and should be promptly referred to an ophthalmologist, because early presentation of patient in the initial stage of the tumor will result in decreased risk of tumor extension, recurrence and mortality.