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

BACKGROUND/AIMS: Classical M1 macrophage activation exhibits an inflammatory phenotype while alternative M2 macrophage activation exhibits an anti-inflammatory phenotype. We aimed to determine whether there are discriminant patterns of macrophage polarization in Crohn's disease (CD) and intestinal tuberculosis (iTB). METHODS: Colonic mucosal biopsies from 29 patients with iTB, 50 with CD, and 19 controls were examined. Dual colored immunohistochemistry was performed for iNOS/CD68 (an M1φ marker) and CD163/CD68 (an M2φ marker), and the ratio of M1φ to M2φ was assessed. To establish the innate nature of macrophage polarization, we analyzed the extent of mitochondrial depolarization, a key marker of inflammatory responses, in monocyte-derived macrophages obtained from CD and iTB patients, following interferon-γ treatment. RESULTS: M1φ polarization was more prominent in CD biopsies (P=0.002) than in iTB (P=0.2) and control biopsies. In granuloma-positive biopsies, including those in CD, M1φ predominance was significant (P=0.001). In iTB, the densities of M1φ did not differ between granuloma-positive and granuloma-negative biopsies (P=0.1). Interestingly, higher M1φ polarization in CD biopsies correlated with high inflammatory response exhibited by peripheral blood-derived monocytes from these patients. CONCLUSIONS: Proinflammatory M1φ polarization was more common in colonic mucosa of CD patients, especially in the presence of mucosal granulomas. Further characterization of the innate immune system could help in clarifying the pathology of iTB and CD.
Biopsy ; Colon ; Crohn Disease* ; Granuloma ; Humans ; Immune System ; Immunohistochemistry ; Macrophage Activation ; Macrophages* ; Monocytes ; Mucous Membrane ; Pathology ; Phenotype ; Tuberculosis*

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.