miR-188-3p-targeted Regulation of ATG7 Affects Cell Autophagy in Patients With Non-obstructive Azoospermia

Abstract Background: Studies have found that ATG7 knockout mice have defects in acrosome development, and the sperm formed by them are similar to human round-headed sperm. miR-188-3p expression is alleviated in the testis tissues of NOA samples. It is unclear whether ATG7 gene is related to non-obstructive azoospermia and whether miR-188-3p and ATG7 have direct target regulation relationship to affect spermatogenesis.Methods: The testicular tissues of 14 non-obstructive azoospermia (NOA) patients and 13 healthy patients were collected for basic scientific research from October 2017 to June 2018. High-throughput sequencing, qRT-PCR, Western blot, immunohistochemical staining, immunofluorescence, electron microscope, and luciferase experiment were performed to confirm the correlation between miR-188-3p and ATG7 in non-obstructive azoospermia.Results: ATG7 was highly expressed in NOA group. ATG7 protein was localized in the cytoplasm of spermatogenic cells at various levels, and the expression in NOA group was dramatically higher than that of normal individuals. After overexpression of miR-188-3p, the ATG7 3'UTR-WT luciferase activity was impeded; while the ATG7 3'UTR-MUT luciferase activity had no significant difference. LC3 and Beclin-1 gene and protein expression in the NOA samples were elevated in comparison with the normal samples; LC3 was mainly located in the cytoplasm of spermatogenic cells at all levels. LC3 and Beclin-1 genes expression after miR-188-3p overexpression were prominently in comparison with the normal samples; after miR-188-3p was inhibited, LC3 and Beclin-1 expressions were dramatically higher. The degree of LC3 punctate aggregation in the miR-188-3p mimic NC samples was greater, and that in the miR-188-3p inhibitor group was greater. The number of autophagosomes of the miR-188-3p mimic samples was less compared with the mimic NC group.Conclusion: We demonstrated here that the ATG7 gene was located in all levels of spermatogenic cells and was highly expressed in the NOA group. The expression of ATG7 gene was negatively correlated with miR-188-3p that may regulate the target gene ATG7 to participate in autophagy and affect the occurrence of NOA.

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Auditory Selective Attention in Cerebral-Palsied Individuals

Language Speech and Hearing Services in Schools ◽

10.1044/0161-1461.1604.260 ◽

1985 ◽

Vol 16 (4) ◽

pp. 260-266 ◽

Cited By ~ 4

Author(s):

Lee Ann Laraway

Keyword(s):

Selective Attention ◽

White Noise ◽

Normal Subjects ◽

Auditory Selective Attention ◽

Normal Individuals ◽

Significant Difference

The purpose of this study was to determine whether there is a statistically significant difference between the auditory selective attention abilities of normal and cerebral-palsied individuals. Twenty-three cerebral-palsied and 23 normal subjects between the ages of 5 and 21 were asked to repeat a series of 30 items consisting of from 2 to 4 digits in the presence of intermittent white noise. Results of the study indicate that cerebral-palsied individuals perform significantly poorer than normal individuals when the stimulus is accompanied by noise. Noise was not a significant factor in the performance of the normal subjects regardless of age.

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Enrichment of common carp (Cyprinus carpio) diet with Malic acid: Effects on skin mucosal immunity, antioxidant defecne and growth performance

Annals of Animal Science ◽

10.2478/aoas-2020-0092 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Roghieh Safari ◽

Seyed Hossein Hoseinifar ◽

Maryam Dadar ◽

Hien Van Doan

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Malic Acid ◽

Control Group ◽

Skin Mucus ◽

Antioxidant Genes ◽

Treatment Comparison ◽

Genes Expression ◽

Significant Difference ◽

And Control

AbstractThe present study investigated possible effects of dietary malic acid on the expression of immunity, antioxidant and growth related genes expression as well as skin mucus immune parameters in common carp. Common carp (Cyprinus carpio) fingerlings were fed diets supplemented with different levels (0 [control], 0.5%, 1%, 2%) of malic acid (MA) for 60 days. The results revealed highest expression levels of immune-related genes (tnf-alpha, il1b, il8 and lyz) in skin of common carp fed 2% MA (P < 0.05). Regarding 1% MA treatment comparison with control group, significant difference was noticed just in case of lyz (P < 0.05). Evaluation of growth related genes expression revealed no significant difference between treatments (P > 0.05). The study of antioxidant related genes (gsta and gpx) in common carp skin fed with MA, showed significant difference between treated groups and control (P < 0.05). Carps fed with 2% MA had highest alkaline phosphatase activity in skin mucus compared other treated groups and control (P < 0.05). There were no significant difference between 0.5% and 1% and control (P > 0.05). The study of total protein and total immunoglobulin (Ig) in common carp skin musus revealed no alteration following MA treatment (P > 0.05). The present data demonstrated that feeding with MA altered immune and antioxidant genes expression in skin mucus of common carp.

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Decreased expression of DNA methyltransferases in the testes of patients with non-obstructive azoospermia leads to changes in global DNA methylation levels

Reproduction Fertility and Development ◽

10.1071/rd18246 ◽

2019 ◽

Vol 31 (8) ◽

pp. 1386 ◽

Cited By ~ 1

Author(s):

Fatma Uysal ◽

Gokhan Akkoyunlu ◽

Saffet Ozturk

Keyword(s):

Dna Methylation ◽

Male Infertility ◽

De Novo ◽

Biological Effects ◽

Testicular Biopsy ◽

Round Spermatid ◽

Dna Methyltransferases ◽

Global Dna Methylation ◽

Obstructive Azoospermia ◽

Spermatogenic Cells

DNA methylation plays key roles in epigenetic regulation during mammalian spermatogenesis. DNA methyltransferases (DNMTs) function in de novo and maintenance methylation processes by adding a methyl group to the fifth carbon atom of the cytosine residues within cytosine–phosphate–guanine (CpG) and non-CpG dinucleotide sites. Azoospermia is one of the main causes of male infertility, and is classified as obstructive (OA) or non-obstructive (NOA) azoospermia based on histopathological characteristics. The molecular background of NOA is still largely unknown. DNA methylation performed by DNMTs is implicated in the transcriptional regulation of spermatogenesis-related genes. The aim of the present study was to evaluate the cellular localisation and expression levels of the DNMT1, DNMT3A and DNMT3B proteins, as well as global DNA methylation profiles in testicular biopsy samples obtained from men with various types of NOA, including hypospermatogenesis (hyposperm), round spermatid (RS) arrest, spermatocyte (SC) arrest and Sertoli cell-only (SCO) syndrome. In the testicular biopsy samples, DNMT1 expression and global DNA methylation levels decreased gradually from the hyposperm to SCO groups (P<0.05). DNMT3A expression was significantly decreased in the RS arrest, SC arrest and SCO groups compared with the hyposperm group (P<0.05). DNMT3B expression was significantly lower in the RS arrest and SCO groups than in the hyposperm group (P<0.05). Although both DNMT1 and DNMT3A were localised in the cytoplasm and nucleus of the spermatogenic cells, staining for DNMT3B was more intensive in the nucleus of spermatogenic cells. In conclusion, the findings suggest that significant changes in DNMT expression and global DNA methylation levels in spermatogenic cells may contribute to development of male infertility in the NOA groups. Further studies are needed to determine the molecular biological effects of the altered DNMT expression and DNA methylation levels on development of male infertility.

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A proton NMR study of the effect of a new intravasal injectable male contraceptive RISUG on seminal plasma metabolites

Reproduction ◽

10.1530/rep.0.1220431 ◽

2001 ◽

pp. 431-436 ◽

Cited By ~ 33

Author(s):

U Sharma ◽

K Chaudhury ◽

NR Jagannathan ◽

SK Guha

Keyword(s):

Nuclear Magnetic Resonance ◽

Seminal Plasma ◽

Vas Deferens ◽

Plasma Metabolites ◽

Obstructive Azoospermia ◽

Male Contraceptive ◽

Nmr Study ◽

Significant Difference ◽

Styrene Maleic Anhydride ◽

Prostatic Diseases

Nuclear magnetic resonance (NMR) spectroscopy was used to quantify citrate, glucose, lactate, glycerophosphorylcholine and choline in seminal plasma from subjects injected with a new male contraceptive RISUG, a copolymer of styrene maleic anhydride dissolved in dimethyl sulphoxide, and in seminal plasma from normal ejaculates. No significant difference in the concentration of citrate was observed between the groups, indicating that the prostate is not affected by the contraceptive. The concentrations of glucose, lactate, glycerophosphorylcholine and choline were significantly lower (P < 0.01) in subjects injected with RISUG compared with controls. In addition, metabolite ratios such as choline:citrate, citrate:lactate, choline:lactate and glycerophosphorylcholine:choline were calculated. Citrate:lactate and glycerophosphorylcholine:choline ratios were significantly lower in RISUG-injected subjects than in controls (P < 0.01), thereby indicating the occurrence of partial obstructive azoospermia. The most important finding of the present study was that the intervention of RISUG in the vas deferens even for a period as long as 8 years is absolutely safe and does not lead to prostatic diseases.

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Periphyton diversity in two different Antarctic lakes assessed using metabarcoding

Antarctic Science ◽

10.1017/s0954102021000316 ◽

2021 ◽

pp. 1-9

Author(s):

Paulo E.A.S. Câmara ◽

Láuren M.D. De Souza ◽

Otávio Henrique Bezerra Pinto ◽

Peter Convey ◽

Eduardo T. Amorim ◽

...

Keyword(s):

Dna Sequences ◽

High Throughput Sequencing ◽

Shannon Index ◽

South Shetland Islands ◽

King George Island ◽

Deception Island ◽

Maritime Antarctic ◽

Shetland Islands ◽

Antarctic Lakes ◽

Significant Difference

Abstract Antarctic lakes have generally simple periphyton communities when compared with those of lower latitudes. To date, assessment of microbial diversity in Antarctica has relied heavily on traditional direct observation and cultivation methods. In this study, sterilized cotton baits were left submerged for two years in two lakes on King George Island and Deception Island, South Shetland Islands (Maritime Antarctic), followed by assessment of diversity by metabarcoding using high-throughput sequencing. DNA sequences of 44 taxa belonging to four kingdoms and seven phyla were found. Thirty-six taxa were detected in Hennequin Lake on King George Island and 20 taxa were detected in Soto Lake on Deception Island. However, no significant difference in species composition was detected between the two assemblages (Shannon index). Our data suggest that metabarcoding provides a suitable method for the assessment of periphyton biodiversity in oligotrophic Antarctic lakes.

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Unravelling the developmental and functional significance of an ancient Argonaute duplication

10.1101/2020.02.04.933887 ◽

2020 ◽

Cited By ~ 1

Author(s):

Arie Fridrich ◽

Vengamanaidu Modepalli ◽

Magda Lewandowska ◽

Reuven Aharoni ◽

Yehu Moran

Keyword(s):

Messenger Rna ◽

High Throughput Sequencing ◽

Genetic Manipulation ◽

Sea Anemones ◽

Differential Distribution ◽

Animal Physiology ◽

Rna Targets ◽

Target Regulation ◽

Bilaterian Animal ◽

Almost All

AbstractmicroRNAs (miRNAs), base-pair to messenger RNA targets and guide Argonaute proteins to mediate their silencing. This target regulation is considered crucial for animal physiology and development. However, this notion is based exclusively on studies in bilaterians, which comprise almost all lab model animals. To fill this glaring phylogenetic gap, we characterized the functions of two Argonaute paralogs in the sea anemone Nematostella vectensis of the phylum Cnidaria, which is separated from bilaterians by ∼600 million years. Using genetic manipulation, Argonaute-immunoprecipitations and high-throughput sequencing we provide experimental evidence for the developmental importance of miRNAs in a non-bilaterian animal. Additionally, we uncover unexpected differential distribution of distinct miRNAs between the two Argonautes and the ability of one of them to load additional types of small RNAs. This enables us to postulate a novel model for evolution of miRNA precursors in sea anemones and their relatives, revealing alternative trajectories for metazoan miRNA evolution.

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Comparison of Oral Hygiene Status and Relative Abundance of Periodontal Pathogens Between Individuals With and Without Intellectual Disabilities: a Cross-Sectional Study

10.21203/rs.3.rs-49931/v1 ◽

2020 ◽

Author(s):

Yuki Oda ◽

Chiaki Furutani ◽

Reo Kawano ◽

Jumpei Murakami ◽

Yuika Mizota ◽

...

Keyword(s):

Periodontal Disease ◽

Relative Abundance ◽

Outpatient Clinic ◽

High Throughput Sequencing ◽

Analysis Of Covariance ◽

University Hospital ◽

Permanent Teeth ◽

Control Group ◽

Periodontal Pathogens ◽

Significant Difference

Abstract Background: Studies have reported a high prevalence of periodontal disease in individuals with intellectual disability (ID). The aim of this study was to compare the relative abundance of periodontal pathogens (red, orange, yellow, purple, and green complexes) between individuals with ID and healthy controls.Methods: Of the 31 subjects enrolled in this study, 16 with severe ID were selected from the outpatient clinic of the Special Care Dentistry of Hiroshima University Hospital, and 14 healthy subjects were selected from the outpatient clinic of another department at the same hospital. Dental plaque was sampled after oral examination. Decayed, missing, and filled permanent teeth (DMFT) counts were obtained and periodontal measurements were taken using the papillary-marginal-attached (PMA) index, plaque index (PI), gingival index (GI), and probing depth (PD). Gene sequencing (16S rRNA) was performed for each sample using next-generation high-throughput sequencing methods. The relative abundance of the periodontal pathogens and the clinical parameters were compared. An unpaired t-test was used to compare the oral health status in both groups and analysis of covariance was performed to compare the relative abundance of each pathogen.Results: No statistically significant difference in DMFT was observed between the two groups. However, significant differences in the median PMA index, PI, and GI were noted between the groups (P < 0.0001). In addition, the mean PD in the ID group was significantly higher than that in the control group (P < 0.0001). The relative abundances of Tannerella spp. and Treponema spp. were significantly higher in the ID group when compared to the control group at the genus level (P = 0.0383 and 0.0432, respectively); alternatively, the relative abundance of Porphyromonas spp. was significantly lower in the ID group (P < 0.0001).Conclusions: The PMA index, PI, GI, and PD were significantly lower in the ID group than in the control group. On the other hand, no significant difference in DMFT was observed between the two groups. Furthermore, our findings indicate that Tannerella forsythia might be more closely associated with periodontal disease than Porphyromonas gingivalis in individuals with ID.

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Anaplasma marginale Superinfection Attributable to Pathogen Strains with Distinct Genomic Backgrounds

Infection and Immunity ◽

10.1128/iai.02537-14 ◽

2014 ◽

Vol 82 (12) ◽

pp. 5286-5292 ◽

Cited By ~ 11

Author(s):

Eduardo Vallejo Esquerra ◽

David R. Herndon ◽

Francisco Alpirez Mendoza ◽

Juan Mosqueda ◽

Guy H. Palmer

Keyword(s):

High Throughput Sequencing ◽

Anaplasma Marginale ◽

Host Population ◽

Content Type ◽

Tropical Regions ◽

Disease Epidemiology ◽

Infectious Disease Epidemiology ◽

Perfect Sequence ◽

Significant Difference ◽

High Prevalence

ABSTRACTStrain superinfection occurs when a second pathogen strain infects a host already infected with a primary strain. The selective pressures that drive strain divergence, which underlies superinfection, and allow penetration of a new strain into a host population are critical knowledge gaps relevant to shifts in infectious disease epidemiology. In regions of endemicity with a high prevalence of infection, broad population immunity develops againstAnaplasma marginale, a highly antigenically variant rickettsial pathogen, and creates strong selective pressure for emergence of and superinfection with strains that differ in their Msp2 variant repertoires. The strains may emerge either bymsp2locus duplication and allelic divergence on an existing genomic background or by introduction of a strain with a differentmsp2allelic repertoire on a distinct genomic background. To answer this question, we developed a multilocus typing assay based on high-throughput sequencing of non-msp2target loci to distinguish among strains with different genomic backgrounds. The technical error level was statistically defined based on the percentage of perfect sequence matches of clones of each target locus and validated using experimental single strains and strain pairs. Testing ofA. marginale-positive samples from tropical regions whereA. marginaleinfection is endemic identified individual infections that contained unique alleles for all five targeted loci. The data revealed a highly significant difference in the number of strains per animal in the tropical regions compared to infections in temperate regions and strongly supported the hypothesis that transmission of genomically distinctA. marginalestrains predominates in high-prevalence areas of endemicity.

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Patterns in the Parathyroid Response to Sodium Bicarbonate Infusion Test in Healthy Volunteers

BioMed Research International ◽

10.1155/2014/704394 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 1

Author(s):

Theodossis S. Papavramidis ◽

Olympia E. Anastasiou ◽

Ioannis Pliakos ◽

Nick Michalopoulos ◽

Mike Polyzonis ◽

...

Keyword(s):

Vitamin D ◽

Sodium Bicarbonate ◽

Healthy Volunteers ◽

Infusion Test ◽

Normal Individuals ◽

Vitamin D Levels ◽

Significant Difference ◽

Low Levels ◽

Bicarbonate Infusion ◽

Sodium Bicarbonate Infusion

Background. The sodium bicarbonate infusion test evaluates the function of the parathyroid glands. The present study aims to evaluate the range of parathyroid response in healthy individuals and the potential influence of various factors.Methods. Fifty healthy volunteers were subjected to the test. Levels of vitamin D, calcium, albumin, and PTH were measured before infusion. PTH was measured at 3, 5, 10, 30, and 60 minutes after infusion.Results. A curve describing the response of parathyroids to the test was drawn. Twenty percent of the subjects had blunted PTH response. No significant difference was observed between normal and blunted responders concerning age, BMI, baseline PTH, or calcium levels. Nonetheless, there was a significant difference in vitamin D levels (P=0.024).Interpretation. The test is easy to perform and may be used for everyday screening. It has to be clarified whether our observations are, at least partly, produced due to the presence of individuals with a constitutively blunted response or if low levels of vitamin D decrease the ability of the parathyroids to respond. Whichever the case, PTH response of normal individuals to sodium bicarbonate infusion test is more varied than previously thought and vitamin D levels influence it.

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INFANTILE SPASTIC HEMIPLEGIA

PEDIATRICS ◽

10.1542/peds.15.6.676 ◽

1955 ◽

Vol 15 (6) ◽

pp. 676-682

Author(s):

Meyer A. Perlstein ◽

Philip N. Hood

Keyword(s):

Close Agreement ◽

Mental Deficiency ◽

Consecutive Series ◽

Selective Population ◽

Normal Individuals ◽

Significant Difference ◽

Spastic Hemiplegia ◽

Left And Right ◽

The Mean ◽

Mental Defectives

From this study the following conclusions emerge regarding the relation of intelligence of infantile spastic hemiplegics to various factors. The spastic hemiplegics of the present series were retarded approximately 20 I.Q. points below normal individuals. The mean I.Q. scores and the incidence of mental retardation were in close agreement with several early reports. The marked difference from the recent reports of Hohman and others reporting consecutive series of cerebral palsied patients is attributed to the fact that the present study is based upon a selective population. Left and right hemiplegics did not differ significantly either in mean I.Q. scores or in the incidence of mental deficiency. Male and female hemiplegics did not differ significantly, either in mean I.Q. scores or in the incidence of mental deficiency. The mean I.Q. scores of the congenital and acquired cases did not differ significantly nor was there a greater incidence of mental deficiency in either of the 2 groups. Mental deficiency was significantly related to the presence of convulsions in this series of spastic hemiplegics; the mean I.Q. score of the convulsives was significantly lower than that for the nonconvulsives by 12 I.Q. points. There was a significantly greater number of mental defectives below the age of 6 than above that age, although the mean I.Q. scores did not differ significantly between the 2 groups. When the subjects were grouped into age levels of 1-year intervals, there was no greater incidence of mental deficiency nor did the mean I.Q. scores differ significantly at any year level. The infectious and traumatic groups of acquired etiologies showed no significant difference in mean I.Q. scores. The toxemia of pregnancy group showed approximately the same incidence of mental deficiency and the same mean I.Q. score as the group composed of all other congenital etiologies of spastic hemiplegia.

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