Successful Results of Intracytoplasmic Sperm Injection of a Chinese Patient With Multiple Morphological Abnormalities of Sperm Flagella Caused by a Novel Splicing Mutation in CFAP251

Asthenospermia is one of the most important causes of male infertility. Among asthenospermia, multiple morphological abnormalities of sperm flagella (MMAF) are relatively rare idiopathic conditions characterized by multiple defects in sperm flagella. Although many studies focusing on the genetic factors of MMAF have been conducted, its pathogenesis and treatment effect remain largely unknown. Here, we report a male patient from a nonconsanguineous Chinese family who exhibited a typical MMAF phenotype revealed by morphological analysis. We identified splicing mutations in CFAP251 (c.1192-3C>G), and the mutation was proven to cause exon skipping. In addition, western blotting and immunofluorescence analysis of the spermatozoa from the proband and a control subject revealed a significantly lower expression of CFAP251 protein due to pathogenic mutation. Interestingly, the patient’s mother was a heterozygous carrier for the mutation, but his father was not, and finally, the inheritance pattern was proven to be maternal uniparental disomy. We applied an intracytoplasmic sperm injection and achieved a successful pregnancy. Above all, our findings expand the spectrum of CFAP251 pathogenic mutations and provide more indications for clinical genetic counseling and assisted reproductive treatment for such patients.

Short notes on teratology in Neocollyris (Heterocollyris) similior (Horn, 1893) (Coleoptera: Cicindelidae): Input to morphological systematics

In this paper, we presented a short observation on the morphological abnormalities of female Neocollyris (Heterocollyris) similior (Horn, 1893) which possibly could create confusion especially to the beginning taxonomists or hobbyists who wishes to describe species new to science.

Nevus Lipomatosus Cutaneous Superficialis with a Histopathological Appearance Resembling Sclerotic Fibroma: A Rare Case Report

Background: Nevus lipomatosus cutaneous superficialis (NLCS) of Hoffmann–Zurhelle is a benign idiopathic hamartoma. There are two types of NLCS, multiple and solitary. They are found in the abdomen, lower back, buttocks, hips, upper posterior thighs, and pelvis. The diagnosis can be evaluated with a typical histopathological of mature fat cells in the dermis, with 10%–50% of the dermis. Case Report: We reported a case of NLCS with clinical papules and multiple nodules on the buttocks since the age of 6 years with a history of lipoma removal. The dermoscopic examination was conducted to confirm the diagnosis. The histopathological examination showed a dominant sclerotic fibroma with two sessions of biopsy and a few mature fats on the dermis after deeper cuts paraffin block. Cryotherapy with an open spray method is treatment of choice in this patient. Discussion: The appearance of the dermis in NLCS can be normal or an increase in collagen. Interestingly, collagen has sclerosis partially and resembles sclerotic fibroma never been reported. NLCS increases the amount of collagen; however, collagen as sclerosis remains obscure. The features of NLCS histopathological with other morphological abnormalities in the dermis have been reported, such as NLCS with perifollicular fibroma (PF) features. The sclerotic fibroma features are other morphological abnormalities in NLCS, as reported in the PF.

A 1-bp deletion in bovine QRICH2 causes low sperm count and immotile sperm with multiple morphological abnormalities

Background: Semen quality and male fertility are monitored in artificial insemination bulls to ensure high insemination success rates. Only ejaculates that fulfill minimum quality requirements are processed and eventually used for artificial inseminations. We examined 70,990 ejaculates from 1343 Brown Swiss bulls to identify bulls from which all ejaculates were rejected due to low semen quality. This procedure identified a bull that produced twelve ejaculates with an aberrantly low number of sperm (0.2±0.2 x 109 sperm per ml) which were mostly immotile due to multiple morphological abnormalities. Results: The genome of the bull was sequenced at 12-fold coverage to investigate a suspected genetic cause. Comparing the sequence variant genotypes of the bull with those from 397 fertile bulls revealed a 1-bp deletion in the coding sequence of QRICH2 encoding glutamine rich 2 as a compelling candidate causal variant. The 1-bp deletion causes a frameshift in translation and induces a premature termination codon (ENSBTAP00000018337.1:p.Cys1644AlafsTer52). The analysis of testis transcriptomes from 76 bulls showed that the transcript with the premature termination codon is subjected to nonsense-mediated mRNA decay. The 1-bp deletion resides on a 675 kb haplotype spanning 181 SNPs from the Illumina BovineHD Bead chip. The haplotype segregates at a frequency of 5% in the Brown Swiss cattle population. This analysis also identified another bull that carried the 1-bp deletion in the homozygous state. Semen analyses from the second bull confirmed low sperm concentration and immotile sperm with multiple morphological abnormalities primarily affecting the sperm flagellum and, to a lesser extent, the sperm head. Conclusions: A recessive loss-of-function allele of bovine QRICH2 likely causes low sperm concentration and immotile sperm with multiple morphological abnormalities. Routine sperm analyses unambiguously identify homozygous bulls. A direct gene test can be implemented to monitor the frequency of the undesired allele in cattle populations.

Loss of iqgap1 (IQ Motif Containing GTPase Activating Protein 1) in Zebrafish Leads to Intracerebellar Hemorrhage, Morphological Abnormalities and Activates Hematopoietic Response

Defects in multiple cell signaling molecules lead to disruptions of vascular integrity given the need for fine-tuned regulation of the cell adhesion complexes. These genetic defects have been linked to the development of intracerebral hemorrhage (ICH). There is genetic evidence in humans for ICH due to some genetic variants, while other variants have been identified in preclinical animal models. Signaling adaptor proteins play a crucial role in cell signaling by promoting interactions between effector proteins and even enabling integration of different pathways. IQGAP1 is a conserved signaling adaptor known for its roles in cell adhesion, cancer and for other cell biological effects. We engineered a zebrafish null mutant in the zebrafish iqgap1 gene by introducing an 11-bp deletion using a CRISPR/Cas9 genome editing method and characterized its phenotype. Homozygous mutants exhibit severe brain hemorrhage and morphological abnormalities, which are ultimately lethal, in about 30-40% of cases, whereas the other embryos survive to adulthood. We visualized the expression pattern of iqgap1 relative to the established fli1a vascular marker and found that iqgap1 strongly overlapped with fli1a expression, but was expressed much more broadly in tissues, such as muscle, branchial arches, and the caudal hematopoietic tissue (equivalent to the mammalian fetal liver). Critically, iqgap1 exhibited co-localization with fli1a in the blood vessels of the central nervous system, whose disruption is likely responsible for the brain hemorrhage. Whole embryo RNA sequencing-based comparison of hemorrhage-positive iqgap1-/- embryo pools with wild-type embryos at 52 hours post-fertilization (hpf) shortly after the onset of hemorrhage identified approximately 800 differentially regulated genes. The most striking feature of this dataset was up-regulation of hematopoietic markers especially those of erythrocytes, neutrophils, mast cells and HSPCs (hematopoietic stem and progenitor cells), but not macrophages. We have confirmed by in situ hybridization with marker gene probes that erythrocyte and neutrophil production is up-regulated most strongly in iqgap1-/- embryos undergoing some level of hemorrhage. By contrast, fli1a endothelial and stem cell marker was downregulated. This animal model provides a compelling genotype-phenotype correlation, implicating IQGAP1 as a new player in vascular disorders such as ICH and identifying a previously unrecognized relationship between IQGAP and regulation of hematopoiesis. Furthermore, this model is now poised to identify ameliorating and exacerbating modifier lesions and potential therapeutic agents that restore normal vascular integrity and prevent ICH. Disclosures Robitaille: Novartis: Consultancy. Berman: Oxford Immune Algorithmics: Membership on an entity's Board of Directors or advisory committees.

Eleven Mutations in Six Genes Causing Abnormal Sperm Morphology: A Case Series

Objective: Infertility has become a major health problem worldwide, approximately 10%–15% of male infertility cases can be attributed to genetic mutations. However, the genetic factors contributing to male infertility remain largely unknown. This study aimed to identify genetic mutations associated with morphological abnormalities in sperm.Methods: In this study, we assessed the semen parameters of 60 patients with sperm morphological abnormalities by semen analysis, and subsequently performed targeted next-generation sequencing (NGS) to identify gene mutations. Results: Of the 60 cases of male infertility, 8 individuals (13%) were identified as carrying genetic mutations. In brief, patient 1 carried two heterozygous mutations of SUN5, including c.1066C>T and c.638delC; Patient 2 and Patient 3 had a 109.73 kb homozygous deletion on chromosome 12 in DPY19L2; patient 4 presented two heterozygous mutations (c.1670_1671delinsTT and c.255_258del) of DPY19L2; patient 5 carried a homozygous variant c.1159+1G>A of CCDC40; patient 6 had a heterozygous mutation c.38_43del in the SEPT12 gene; patient 7 had two heterozygous mutations in DNAH1 (c.10060_10061insATCT and c.12278T>C), and patient 8 presented two heterozygous mutations of CFAP43 (c.3658C>T and c.823del). Conclusions: We identified gene mutations associated with sperm morphological abnormalities in a series of male infertility cases. Eight of sixty individuals were identified as carrying gene mutations. Genetic mutations of SUN5, DPY19L2, CCDC40, SEPT12, DNAH1, and CFAP43 might be responsible for male infertility. Our study expanded the landscape of gene mutations related to sperm morphological abnormalities, which might provide new insights into the genetics underlying male infertility.