Sperm retrieval rate and reproductive outcome of infertile men with azoospermia factor c deletion.

To evaluate the success rate in sperm retrieval (SR) through microdissection testicular sperm extraction (micro-TESE) in infertile azoospermia factor c (AZFc)-deleted men and determining their reproductive outcomes following ICSI, medical records of couples with AZFc-deleted male partners were reviewed on patient's age, serum hormone levels, karyotype, testicular pathology and pregnancy outcomes. A comparison on age and serum hormone level was conducted between groups with positive and negative sperm retrieval in both azoospermic and oligozoospermic AZFc-deleted men. Of 225 who had AZFc deletion, 195 cases followed clinical treatments. From 195 cases, 116 were azoospermic, 79 were oligozoospermic. Pathology profile was available in 103 of 195 subjects which the predominant trait was SCOS and was seen in 66.9% of cases (69 of 103). Success rate of sperm retrieval in azoospermic patients who underwent micro-TESE was 36.3% (28/77). Forty-three oligozoospermic and 17 azoospermic patients started ART cycle. Pregnancy rate in oligozoospermic group was 35.4% (17 cases), whilst there was no clinical pregnancy in azoospermic group. In conclusion, the pregnancy and delivery in oligozoospermic patients with AZFc deletion are comparable with other studies, but despite of sperm retrieval in azoospermic patients with AZFc deletion, the chance of pregnancy or delivery in these patients was very low.

Numerical simulation to optimize power conversion efficiency of an FTO/GO/Cs2AgBiBr6/Cu2O solar cell.

Efficient conversion of solar power to electrical power through the development of smart, reliable, and environmentally friendly materials is a key focus for the next-generation renewable energy sector. The involvement of degradable and toxic elements present in hybrid perovskites presents serious concerns regarding the commercial viability of these materials for the solar cell industry. In this study, a solar cell with a stable, nondegradable, and lead-free halide-based double perovskite Cs2AgBiBr6 as the absorber layer, Cu2O as a hole transport layer, and GO as the electron transport layer has been simulated using SCAPS 1D. The thickness of the absorber, electron transport, and hole transport layers are tuned to optimize the performance of the designed solar cell. Notably, perovskite solar cells functioned most efficiently with an electron affinity value of 4.0 eV for Cu2O. In addition, the effect of variation of series resistance and temperature on generation and recombination rates, current density, and quantum efficiency has been elaborated in detail. The findings of this study provide valuable insight and encouragement toward the realization of a non-toxic, inorganic perovskite solar device and will be a significant step forward in addressing environmental concerns associated with perovskite solar cell technology.

Specific Deoxyceramide Species Correlate with Expression of Macular Telangiectasia Type 2 (MacTel2) in a SPTLC2 Carrier HSAN1 Family.

Hereditary sensory and autonomic neuropathy type 1 (HSAN1/HSN1) is a peripheral neuropathy most commonly associated with pathogenic variants in the serine palmitoyltransferase complex (SPTLC1, SPTLC2) genes, which are responsible for sphingolipid biosynthesis. Recent reports have shown that some HSAN1 patients also develop macular telangiectasia type 2 (MacTel2), a retinal neurodegeneration with an enigmatic pathogenesis and complex heritability. Here, we report a novel association of a SPTLC2 c.529A>G p.(Asn177Asp) variant with MacTel2 in a single member of a family that otherwise has multiple members afflicted with HSAN1. We provide correlative data to suggest that the variable penetrance of the HSAN1/MacTel2-overlap phenotype in the proband may be explained by levels of certain deoxyceramide species, which are aberrant intermediates of sphingolipid metabolism. We provide detailed retinal imaging of the proband and his HSAN1+/MacTel2- brothers and suggest mechanisms by which deoxyceramide levels may induce retinal degeneration. This is the first report of HSAN1 vs. HSAN1/MacTel2 overlap patients to comprehensively profile sphingolipid intermediates. The biochemical data here may help shed light on the pathoetiology and molecular mechanisms of MacTel2.

First principles investigation of halide based Rb2NaGaZ6 (Z = Br, I) double perovskites for energy harvesting applications.

Extensive investigations have been conducted on the thermoelectric and optoelectronic characteristics of double perovskite compounds using the full potential linearized augmented plane wave (FP-LAPW) approach. Here we investigated Rb2NaGaZ6 (Z = Br, I) to explore its band structure, and electronic, optical and transport properties. Born's stability criteria have confirmed the mechanical stability of these compounds. Analysis of the elastic properties reveals their ductile nature, as indicated by a Poisson coefficient (υ) greater than 0.26 and a Pugh ratio exceeding 1.75 for Rb2NaGaZ6 (Z = Br, I). Computation of the bandgap values shows that both compositions possess a direct bandgap nature, with respective values of 2.90 eV and 1.25 eV. This suggests that substituting Br with I brings the band edges closer together, resulting in a decrease in the bandgap value. The optical properties are assessed based on the absorption coefficient, reflectivity, and dielectric constants. The thermoelectric properties, including thermal and electrical conductivities, power factor (PF), and figure of merit (ZT), are determined using the BoltzTrap code. The ZT values indicate that both compositions exhibit promising potential for various transportation applications.

VEGF gene mRNA expression in patients with coronary artery disease.

To assess the expression of vascular endothelium growth factor (VEGF) mRNA in unstimulated peripheral blood mononuclear cells of patients with and without coronary artery disease (CAD). We also studied whether the functional VEGF -2,578C/A polymorphism may influence the level of VEGF mRNA expression in individuals undergoing coronary angiography because chest pain. We assessed 50 consecutive patients with angiographically confirmed CAD (CAD+). Also, 50 consecutive individuals with normal coronary studies were included in the study for comparison. VEGF mRNA expression was examined using quantitative real-time PCR and genotyping for VEGF -2,578C/A was performed using ARMS-PCR technique. VEGF mRNA expression was significantly decreased in CAD+ patients when compared to CAD- individuals (p = 0.01). The frequency of VEGF -2578 allele C and genotype CC was increased in CAD+ patients. In this regard, homozygosity for the CC genotype was more commonly observed in CAD+ (30 %) than in those without CAD disease (18 %). However, the difference was slightly out of the range of significance (p = 0.1). In addition, a trend for reduction in the expression of VEGF mRNA was observed when patients carrying the VEGF -2,578AA genotype were compared with those VEGF -2,578AC heterozygous or those homozygous for the VEGF -2,578CC genotype. VEGF gene expression is decreased in individuals with CAD+ disease. The VEGF -2,578C/A polymorphism may influences the expression of VEGF.

Principles and Applications of CRISPR Toolkit in Virus Manipulation, Diagnosis, and Virus-Host Interactions.

Viruses are one of the most important concerns for human health, and overcoming viral infections is a worldwide challenge. However, researchers have been trying to manipulate viral genomes to overcome various disorders, including cancer, for vaccine development purposes. CRISPR (clustered regularly interspaced short palindromic repeats) is becoming one of the most functional and widely used tools for RNA and DNA manipulation in multiple organisms. This approach has provided an unprecedented opportunity for creating simple, inexpensive, specific, targeted, accurate, and practical manipulations of viruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), human immunodeficiency virus-1 (HIV-1), and vaccinia virus. Furthermore, this method can be used to make an effective and precise diagnosis of viral infections. Nevertheless, a valid and scientifically designed CRISPR system is critical to make more effective and accurate changes in viruses. In this review, we have focused on the best and the most effective ways to design sgRNA, gene knock-in(s), and gene knock-out(s) for virus-targeted manipulation. Furthermore, we have emphasized the application of CRISPR technology in virus diagnosis and in finding significant genes involved in virus-host interactions.

Demyelination in hereditary sensory neuropathy type-1C.

OBJECTIVE: Sphingolipids are enriched in the nerves. Serine-palmitoyltransferase (SPT) catalyzes the key step of sphingolipids biosynthesis. Mutations in SPT subunits (SPTLC) lead to the excessive production of neurotoxic deoxysphingolipids (DoxSLs) in patients with Hereditary Sensory Neuropathy Type-1C (HSN1C). HSN1C is an autosomal dominant peripheral neuropathy characterized by sensory loss and distal muscle weakness. In this study, by leveraging a HSN1C family with a previously reported N177D mutation in SPTLC2, we aim to further define the spectrum of DoxSL species and the peripheral neve pathology of the disease. METHODS: Next-generation sequencing along with Sanger confirmation was performed for family members and healthy controls. LC-MS was used for lipidomic analysis in participants' plasma. Quantitative magnetic resonance imaging (qMRI) was performed to study sciatic nerve pathologies. RESULTS: A heterozygous N177D mutation in SPTLC2 was co-segregated in individuals with sensory-motor deficits in the limbs. Nerve conduction studies (NCS) revealed nonuniform slowing of conduction velocities. In line with the NCS, qMRI detected a pattern of nerve changes similar to those in acquired demyelinating polyneuropathies. Additionally, we detected a significant increase in multiple species of deoxysphingoid bases and deoxyceramides in patients' plasma. INTERPRETATION: Mutations in the SPTLC2 cause a demyelinating phenotype resembling those in acquired demyelinating polyneuropathy. The species of increased DoxSLs in HSN1C may be more diverse than originally thought.

Fatigue in patients with hereditary neuropathy with liability to pressure palsies.

OBJECTIVE: Hereditary Neuropathy with Liability to Pressure Palsies (HNPP) is caused by a heterozygous deletion of peripheral myelin protein-22 (PMP22) gene resulting in focal sensorimotor deficits. Our lab has identified a disruption of myelin junctions in excessively permeable myelin that impairs action potential propagation. This mechanism is expected to cause fatigue in patients with HNPP. Therefore, the objective was to characterize fatigue in patients with HNPP and determine the relationship of fatigue to nerve pathology, disability, and quality of life. METHODS: Nine females with HNPP participated in a single visit that included genotyping, nerve conduction studies, neurological exam, quantitative magnetic resonance imaging, and a physical therapy exam incorporating upper and lower extremity function and survey measures of fatigue. This visit was followed by 2 weeks of ecological momentary assessment (wrist-worn device) that captured fatigue ratings five times per day. RESULTS: Participants demonstrated mild neurological impairment (CMTNS: 5.7 ± 2.8), yet reported high fatigue levels (average fatigue intensity over 2 weeks: 5.9 out of 10). Higher fatigue levels were associated with poorer quality of life and more pain. Higher fatigue was associated with significantly greater distal nerve proton density changes on peripheral nerve MRI, which is in line with hyper-permeable myelin in HNPP. INTERPRETATION: Fatigue is common and severe among patients with HNPP whose disabilities are minimal by conventional outcome measures. Therapeutic interventions targeting fatigue have the potential to improve quality of life and may serve as a robust outcome measure to show longitudinal changes for patients with HNPP.

Expanded genetic insight and clinical experience of DNMT1-complex disorder.

OBJECTIVE: To report novel causal mutations, expanded clinical phenotypes, and clinical management of DNA methyltransferase 1 (DNMT1)-complex disorder. METHODS: Neurophysiologic testing, imaging, and genetic findings were summarized in clinical context for 5 cases with DNMT1-complex disorder. RESULTS: We identified 2 novel DNMT1 mutations (p.E510K and p.P1546A) by whole-exome sequencing (WES). Case 1 (p.E510K) presented with childhood ataxia, treatment-refractory seizures, and rapid cognitive decline in his 50s. Case 2 also had childhood onset and presented with seizures, language regression, hearing loss, narcolepsy with cataplexy symptoms, optic atrophy, sensory neuropathy, and hypogammaglobulinemia requiring IV immunoglobulin. Case 2 (p.P1546A) was identified with a de novo and the first mutation residing outside the targeting sequence domain. Case 3 (p.A570V) had paralytic asymmetric onset attacks triggered by emotionality and lasting sometimes for weeks. Neuropsychological testing showed executive dysfunction localizing to frontosubcortical and frontoparietal structures. He gradually developed left predominant brain atrophy. MRI showed T2 hyperintense lesions that enhanced on T1 postgadolinium images, and brain PET showed hypometabolism in atrophied regions. Case 4 (p.T497P) underwent left cochlear implant, resulting in significant hearing improvements at all tested frequencies (250-6,000 Hz). Case 5 (p.Y511H) had profound gait ataxia with posterior column atrophy of the spinal cord and abnormal evoked potentials primarily affecting the fasciculus gracilis. CONCLUSIONS: Broader application of WES further expands genotype-phenotype correlations of DNMT1-complex disorder. Two mutations are identified with early childhood onsets. The expanded new phenotypes include asymmetric brain hemiatrophy with parenchymal gadolinium enhancement, spinal cord atrophy, prolonged cataplectic spells, and hypogammaglobulinemia. Hearing loss treatment by cochlear implantation is helpful and should be considered.

Association between epstein barr virus and tongue squamous cell carcinoma in iranian patients.

Detection of Epstein-Barr virus in oral squamous cell carcinoma suggests its involvement in the carcinogenesis of oral cavity. But, there are few studies on the incidence of EBV genome in squamous cell carcinomas at specific locations in the oral cavity like tongue and with different tumor progression. In this study the presence of EBV genome in tongue Squamous Cell Carcinoma (TSCC) in Iranian patients were investigated. Accordingly, a total of 94 cases with TSCC were firstly analyzed for the presence of viral genome through Nested PCR. Patients were divided into different groups based on their gender and the size, nodal involvement, grade and stage of their tumor. Results showed the presence of EBV genome in 72.3% of TSCCs with no significant difference between two genders, although slightly higher in females. Interestingly, PCR products of EBV genome showed a statistically significant higher distribution in TSCCs at IVa stage (p=0.04), while a considerable low involvement of EBV genome was seen in T1-sized tumors. The result of this study further emphasizes the role of EBV in oral SCCs - mainly at tongue. This is the first investigation to clarify the association between EBV genome and different tumor size and stage in TSCCs; however, more studies in different regions and larger populations should be performed to be able to draw a firmed conclusion.

A Comparative Study of the Impact of Calcium Ion on Structure, Aggregation and Chaperone Function of Human αA-crystallin and its Cataract- Causing R12C Mutant.

BACKGROUND: The chaperone activity of α-crystallin (α-Cry) plays an important role in maintenance of eye lens transparency. Various mutations in the α-Cry genes have been indicated to cause cataract diseases in human. Also, the calcium imbalance has been shown to induce aggregation in α-Cry. We investigated the impact of calcium ion on structure, chaperone activity of the recombinant wild-type and mutant R12C αA-Cry. We suggested that the raise of calcium level in eye lens is an additional contributory factor accelerating the development of cataract diseases in patients with R12C mutation. OBJECTIVES: The main objective of this study was to investigate the impact of calcium ion on structure, chaperone activity and amyloidogenic properties of the recombinant wild-type and mutant R12C αA-Cry, in a comparative study. METHODS: The mutagenesis was performed on confirmed αA-Cry cDNA in pET-28b (+) which applied as a template to generate R12C mutant, using polymerase chain reaction (PCR) and a Quick Change Lightning Multi Site-Directed Mutagenesis kit (Stratgene). Both wild-type and mutant plasmids were chemically transformed into E.coli BL21 (DE3) and the respective recombinant proteins over-expressed in LB broth. The protein purification was done using Q-Sepharose anion exchange and Sephacryl S-300 gel filtration chromatography. The purified αA-Cry samples were incubated with different concentrations of calcium ion (0-40 mM) at 37 °C for 1 week. The secondary and tertiary structural analyses of each protein were performed by far-UV CD and Try/Trp and ANS fluorescence assessments, respectively. The assessment of chaperone activity was done spectrophotometrically in both thermal and chemical-induced aggregation systems using γ-Cry and bovine pancreatic insulin as the substrate proteins, respectively. Also, the amyloidogenic properties of proteins was investigated by CR absorption and ThT fluorescence measurements. RESULTS: The results of fluorescence and CD assessments suggested the significant secondary and tertiary structural alterations upon R12C mutation. R12C mutant αA-Cry demonstrated preserved secondary and tertiary structures in the presence of calcium. The chaperone activity of wild-type and mutant R12C αA-Cry was reduced in the presence of calcium. Also, the extent of chaperone activity reduction was significantly higher for R12C αA-Cry. Both wild-type and mutant R12C αA-Cry revealed slight amount of aggregation when incubated with different calcium concentrations for 1 week, at 37 °C. However, the susceptibility of both proteins for aggregation was significantly increased in the presence of 40 mM calcium, at the elevated temperature (60 °C). Also, the mutant protein exhibited extensive disulfide bridge cross-linking as indicated by gel electrophoresis. Moreover, the mutant R12C αA-Cry significantly resists against amyloid fibril formation in the presence of calcium ion compared to the wild-type protein as indicated by CR and ThT assessments. CONCLUSION: Our data suggested that αA-Cry conformational changes occurring upon R12C mutation and further functional damages induced by calcium may play an important role in the pathomechanism of the cataract development by this mutant protein.

Detection of KCNJ11 gene mutations in a family with neonatal diabetes mellitus: implications for therapeutic management of family members with long-standing disease.

BACKGROUND: Activating mutations of potassium inwardly-rectifying channel, subfamily J, member 11 (KCNJ11), which encodes Kir6.2 (beta-cell adenosine triphosphate-sensitive potassium [K(ATP)] channel subunit), have been associated with neonatal diabetes mellitus (NDM) in different studies. Treatment with oral sulfonylureas in place of exogenous insulin injections results in improved glycemic control in most patients carrying these mutations. Exploration of genetic causes of NDM occurring before the age of 6 months has been proposed as an important issue in identification of monogenic forms of diabetes, which might be critical in their therapeutic management, as a consequence. METHODS: Mutation screening of the KCNJ11 gene was carried out using PCR amplification followed by direct sequencing in three family members: the proband, ND1, diagnosed at 40 days of age (current age 7 years); his sibling, ND2, diagnosed at 2 years of age (current age 14 years); and their father, ND3, diagnosed at 15 years of age (current age 35 years), who had been exclusively treated with insulin. The effect of the E227K mutation was also examined in a homology model of Kir6.2. RESULTS: Our results revealed the presence of the heterozygous missense mutation c. 679 G/A (E227K) in all three patients, who were all able to successfully transfer from insulin injections to an oral sulfonylurea, with improved glycemic control. CONCLUSION: We found that three members of a family with highly variable age of onset of insulin-treated diabetes, diagnosed at 40 days, 2 years, and 15 years of age, all carried the E227K mutation in KCNJ11 and could switch to an oral sulfonylurea. This mutation has been previously reported in patients with permanent and transient NDM, as well as later-onset diabetes; this report adds to the variability in phenotypic presentation and further supports genetic testing in all diabetic members of any family affected by NDM.