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

Aim: We designed this study to compare the presence of depressive symptoms pre and 6 months post-MI and to identify the association of various biological and non-biological risk factors with the development of depressive symptoms post-MI. Objective: Coronary artery disease and depressive illness are the largest contributors to global mortality and disability. These disorders frequently occur together and have major health implications. We conducted this study to determine the frequency of increased depressive symptoms following Myocardial Infarction (MI) and analyze associated baseline risk factors. Study Design: Descriptive case series. Place and duration of study: Tertiary care cardiology hospital from Sept 2019 to Jan 2021. Patients and Methods: Patients presenting with a recent episode of myocardial infarction at the out-patient department of cardiology were recruited for this study. Demographic variables and baseline health status were recorded. Hospital Anxiety and Depression Scale (HADS) was applied in two instances, at first contact for depressive symptoms pre-MI and at the second instance 6 months post-MI. An increase in HADS score of 4 or more points after 6 months of MI was considered significant. Association of age, gender, smoking status, employment status, previous MI episode, depression and history of stressful life event with an increase in depression was analyzed. Results: The sample consisted of 140 patients, 88 males and 52 females. The mean age was 51.22 years (SD= ± 12.35). 60.7% (n=85) of patients had an increase in depressive symptoms scores as measured by HADS. Younger age (30 to 50 years), being a smoker and having a previous history of myocardial infarction were associated with a significant increase in depressive symptoms. Conclusion 60.7% of patients have increased depressive symptoms after myocardial infarction. Patients should be regularly screened for emerging depressive symptoms and special attention should be paid to younger patients, smokers and those who have a previous history of coronary artery disease.

Opioid drugs are the first line of defense in severe pain but the adverse effects associated with opioids are considered as a serious issue worldwide. Acupuncture/electroacupuncture is a type of Chinese medicine therapy which is an effective analgesic therapy, well documented in animals and human studies. Electroacupuncture stimulation could release endogenous opioid peptides causing analgesia in a variety of pain models. It can be used as an alternative therapy to control the opioid crisis.

Objective To report the indigenous people's uses of plants from a multidimensional perspective in a remote area where strong ethnobotanical cultural practices prevail. Methods An ethnobotanical survey was conducted in the field during 2014–2016. Ethnobotanical data were gathered from 182 informants through oral interviews and semi-structured questionnaires. The distribution of plants was explored using both descriptive and graphical methods. Further, a Multinomial Logit Specification was applied to find out the probability of the occurrence of diverse utilization of plants in multipurpose domains. Results The study identified 202 plant species distributed among 71 families and 156 genera. Ethnobotanical data indicate that there are more medicinal (36.96%) uses of plants as compared to all other use categories. The output from the Multinomial Logit Specifications (MLS) model reveals that perennial and non-woody plants are exploited more for medicinal and food uses than annual and woody plants. In the context of ethnomedicinal uses, aerial plant parts particularly leaves are more extensively used for the preparation of herbal recipes as compared to underground parts. Conclusions The results of the study emphasize the need to create awareness among the local communities about the conservation status of plant species in order to maintain a sustainable resource of plant-derived materials into the future. The novel econometric approach employed in this study adds a new insightful methodology to the existing body of literature in the field of ethnobotany. We strongly recommend conservation measures, alongside phytochemical and pharmacological studies on the useful plant species identified in this study in order to ensure their sustainable and effective utilization.

Objective: To document the medicinal and other folk uses of native plants of the area with a view to preserve the ethnobotanical knowledge associated with this area. Methods: The fieldwork was conducted during a period of one year. Data were collected through a semi-structured questionnaire and interviews with indigenous tribal people and traditional health practitioners residing in the study area. Results: The present study documented ethnobotanical uses of 104 plant species belonged to 93 genera and 51 families. Results revealed that most of the documented species were used medicinally (78 spp., 44.07%). Leaves were found to be the most frequently used part (69 spp., 42.86%) for the preparation of indigenous recipes and for fodder. Conclusions: The current research contributes significantly to the ethnobotanical knowledge, and depicts a strong human-plant interaction. There is an urgent need to further document indigenous uses of plants for future domestication.

OBJECTIVE: To document the medicinal and other folk uses of native plants of the Bana Valley of district of Kotli-Azad Jammu Kashmir with a view to preserve the ethnobotanical knowledge of this area and to develop an ethnobotanical inventory of the species diversity. METHODS: The fieldwork was conducted during a period of one year. Data were collected through semi-structured questionnaire and interview of the informants including indigenous people, tribal people, and traditional health practitioners residing in the study area. RESULTS: The present study documented etnobotanical uses of 86 plant species belonging to 81 genera and 47 families. This study revealed that most of the species were used medicinally (74 spp; 42.29%). Leaves found to be the most frequently used part (56 spp., 36.13%) for preparation of indigenous recipes and fodder purpose. CONCLUSIONS: The current research provides a huge lump of ethnobotanical knowledge and depicts strong human-plant interaction. It is an urgent need to document indigenous uses of plants for future domestication.

There are 300 known Saussurea species. Among them, Saussurea lappa (S. lappa) is a representative perennial herb, globally distributed across Himalaya region. S. lappa has been traditionally used in medicines without obvious adverse effects. Despite significant progress in phytochemical and biological analyses of S. lappa over the past few years, inclusive and critical reviews of this plant are anachronistic or quite limited in scope. The present review aims to summarize up-to-date information on the active constituents, pharmacology, traditional uses, trade and challenges in conservation and sustainable use of S. lappa from the literature. In addition to botanical studies and records of the traditional use of S. lappa in over 43 diseases, scientific studies investigating the latent medicinal uses of this species and its constituent phytochemicals for a range of disorders are presented and discussed. The structure, bioactivity, and likely mechanisms of action of S. lappa and its phytochemicals are highlighted. Although some progress has been made, further scrupulous efforts are required to investigate the individual compounds isolated from S. lappa to validate and understand its traditional uses and develop clinical applications. The present review offers preliminary information and gives direction for further basic and clinical research into this plant.

Objective: To report the patterns of species diversity, and regenerating capacity in the forest of Nikyal valley in relation to environmental variables and underlying anthropogenic influence. Methods: A study area was selected in a traditionally managed mountain woody pasture during July 2012 to June 2013. The area was sampled by quadrat method. The quadrats were laid down at regular intervals of 150 m. The size of quadrats was kept 10×10, 5×5 and 1×1 m