Analysis Of Differential Profiles Research Articles

(063) The Role of Leptin as a Testicular Immune Microenvironment Modulator and Leydig Stem Cell Function

Abstract Introduction The testicular microenvironment, consisting of cells including Sertoli and peritubular cells, releases paracrine factors that may regulate testicular function. In previous research, Leptin was identified as a critical paracrine factor in the testicular microenvironment. Leptin is secreted by Sertoli and peritubular cells and binds to its receptor in Leydig cells. This interaction triggers the expression of the desert hedgehog signaling pathway, which is essential for Leydig stem cell differentiation into adult Leydig cells, which are capable of producing testosterone. However, our earlier study had a limitation: the human cell model used was derived from testes biopsies, lacking an intact immune microenvironment. This study investigates the effect of Leptin on a complete testicular immune microenvironment in a murine model. Objective To investigate the impact of Leptin on a complete testicular immune microenvironment. Methods We used 6-week-old C57BL6 mice, exposed to two different concentrations of Leptin, 10 and 100 mg/ml, for seven days. After the treatment period, the animals were humanely euthanized, and their blood and testes were collected. Blood samples underwent a complete blood count differential profile analysis using a Hemavet Mascot Multispecies Hematology System Counter 1500R. Testes samples were subjected to comprehensive immunophenotyping, allowing us to assess the impact of Leptin treatment on various immune cell types within the testes Results Leptin at a low dose (10 mg/ml) showed increased, but not significant, levels of serum testosterone (13.20 + 0.37 ng/dl), while LH and FSH levels significantly (p<0.05) increased. In contrast, animals receiving 100 mg/ml Leptin showed no significant changes in hormone levels. CBC profiling indicated that lymphocytes and platelets were significantly influenced, specifically with low doses of Leptin. Comprehensive immunophenotyping revealed a significant increase in CD4+ (helper T cells) and CD8+ (cytotoxic T lymphocytes) cell numbers in animals treated with low doses of Leptin, a pattern not observed with high doses. Conclusions This study uncovers previously unexplored aspects of how paracrine factors, released by Sertoli and peritubular myoid cells, can modulate the immune microenvironment within the testis, potentially impacting testosterone production. Future research will investigate the relationship between changes in immune cell types, increased T cell numbers, and their influence on Leydig stem cell differentiation into adult Leydig cells Disclosure No.

(251) Exogenous Leptin Treatment Augments Testicular T-cell Secretion and Function Differentially in Context to BMI in Murine Models

Abstract Introduction In previous research, we explored the effects of exogenous Leptin treatment on modulating the immune landscape within the testicular microenvironment (TME). We found that within the TME, it is the CD8+ T cells which were enriched upon Leptin treatment. However, our earlier study did not took into account the impact of variations in BMI which could potentially confound the effects of Leptin on T cells and on TME as a whole. Objective To investigate the impact of Leptin on a complete testicular immune microenvironment. Methods We used 6-week-old C57BL6 mice (n=30), grouped into three, with first group subjected to regular diet, second to Lean fat and third to high fat inducing diet. Once the mice reached the desired weights, they were exposed to two different concentrations of Leptin, 10 and 100 mg/ml, for seven days. After the treatment period, the animals were humanely euthanized, and their blood and testes were collected. Blood samples underwent a CBC differential profile analysis using a Hemavet Mascot Multispecies Hematology System Counter 1500R. Testes samples were subjected to comprehensive immunophenotyping, allowing us to assess the impact of Leptin treatment on various immune cell types within the testes. Results In mice with regular BMI, Leptin at a low dose (10 mg/ml) showed increased, but not significant, levels of serum testosterone (13.20 + 0.37 ng/dl), while LH and FSH levels significantly (p<0.05) increased. In contrast, animals receiving 100 mg/ml Leptin showed no significant changes in hormone levels. CBC profiling indicated that lymphocytes and platelets were significantly influenced, specifically upon low doses of Leptin. Comprehensive immunophenotyping revealed a significant increase in CD4+ (helper T cells) and CD8+ (cytotoxic T lymphocytes) cell numbers in animals treated with low doses of Leptin, a pattern not observed with high doses. These results were in contrast to the mice which represented lean and obese conditions. Conclusions This study uncovers previously unexplored aspects of how exogenous Leptin treatments can modulate the immune microenvironment within the testis, potentially impacting testosterone production. Future research will investigate the relationship between changes in immune cell types, increased T cell numbers, and their influence on Leydig stem cell differentiation into adult Leydig cells. Disclosure No.

Differential profiling analysis of miRNAs reveals a regulatory role in low N stress response of Populus.

Nitrogen (N) is an essential mineral element for plant growth processes, and its availability severely affects the productivity of plants, especially trees. MicroRNAs (miRNAs) are a class of non-coding RNAs approximately 21 nucleotides in length that play important roles in plant growth, development and stress responses. To identify Populus miRNAs and their functions in response to nutrition stress, high-throughput sequencing was performed using Populus tomentosa plantlets treated with or without low concentrations of N. We identified 160 conserved miRNAs, 15 known but non-conserved miRNAs, 2 candidate novel miRNAs and 71 corresponding miRNA*s. Differential expression analysis showed that expression of the 21 conserved miRNA families was significantly altered. Real-time quantitative PCR (qPCR) was used to further validate and analyze the dynamic expression of the identified miRNAs. A total of 218 target genes from the low-N-responsive miRNAs were predicted, and their functions were further annotated in combination with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. These results suggest that miRNAs play important roles in the response of Populus to low N stress. Furthermore, this study provides the first identification and profiles of N stress-responsive miRNAs from trees.

Profiling of Ubiquitin-like Modifications Reveals Features of Mitotic Control

Ubiquitin and ubiquitin-like (Ubl) protein modifications affect protein stability, activity, and localization, but we still lack broad understanding of the functions of Ubl modifications. We have profiled the protein targets of ubiquitin and six additional Ubls in mitosis using a functional assay that utilizes active mammalian cell extracts and protein microarrays and identified 1,500 potential substrates; 80-200 protein targets were exclusive to each Ubl. The network structure is nonrandom, with most targets mapping to a single Ubl. There are distinct molecular functions for each Ubl, suggesting divergent biological roles. Analysis of differential profiles between mitosis and G1 highlighted a previously underappreciated role for the Ubl, FAT10, in mitotic regulation. In addition to its role as a resource for Ubl modifications, our study provides a systematic approach to analyze changes in posttranslational modifications at various cellular states.

Differential profile analysis of urinary cytokines in patients with overactive bladder: reply

Differential profile analysis of urinary cytokines in patients with overactive bladder: reply

Erratum to: Differential profile analysis of urinary cytokines in patients with overactive bladder

Erratum to: Differential profile analysis of urinary cytokines in patients with overactive bladder

Differential profile analysis of urinary cytokines in patients with overactive bladder

We hypothesize that overactive bladder (OAB) can produce inflammatory cytokines due to afferent neural plasticity or urothelial dysfunction. This study aimed to detect abnormal cytokine levels in urine of patients with OAB compared to urinary tract infections (UTI) and controls. This was a prospective, single blind study including 20 premenopausal women (control), 20 with OAB and 16 with UTI. Urine samples were collected, centrifuged, and stored (-80°C). Urinary total proteins were quantified and detected by antibody-based array chip for release of 120 human cytokines in the two groups relative to the controls. Majority of cytokines showed the same expression in the OAB compared with the controls. Cytokines exclusively expressed in OAB were: monocyte chemoattractant protein (MCP) 1, TARC, PARC, and Fas/TNFRSF6. MCP-2, MCP-3, tumor necrosis factor-β, GCSF and eotaxin-3 showed a shared expression in UTI and OAB. Conversely, few of the cytokines were downregulated in OAB (IL-5, IL-6, IL-7, and GM-CSF). Taken together, the results suggest that a subset of inflammatory cytokines and chemokines provides a framework for development of highly optimized urinary biomarker assay for differential diagnosis and treatment of OAB.

FD-LC-MS/MS 法によるタンパク質ディファレンシャルプロファイリング

FD-LC-MS/MS 法によるタンパク質ディファレンシャルプロファイリング

A Proteomics Method Revealing Disease-Related Proteins in Livers of Hepatitis-Infected Mouse Model

In this post-genome era, a sensitive quantitative method is required for differential profiling analyses of clinical proteomes to understand the disease progress. Here, we adopt the FD-LC-MS/MS method, consisting of fluorogenic derivatization (FD), separation by liquid chromatography (LC), and identification by LC-tandem mass spectrometry (MS/MS), to reveal disease-related proteins in livers of hepatocarcinogenesis in transgenic (Tg) and non-transgenic (NTg) mice at three developmental stages. After 6 months, the expression of apoptosis-related proteins is suppressed. After 12 months, proteins related to respiration, the electron-transfer system, and anti-oxidation are significantly up-regulated. After 16 months, proteins related to defense, beta-oxidation, and apoptosis are significantly suppressed. This fluctuating expression of proteins could explain the progression of hepatocarcinogenesis. The method would be useful for clinical proteomics analysis because of its high resolution, sensitivity, and reproducibility.

Coping e gênero em adolescentes

Este estudo avalia a associação entre bem-estar psicológico e estratégias de coping nos gêneros masculino e feminino, bem como o perfil diferencial destas entre os gêneros. A amostra foi composta de 389 jovens, estudantes do terceiro ano do ensino médio de escolas de diferentes regiões de Porto Alegre. Os resultados evidenciam que as meninas, ao assumirem uma postura ativa na resolução dos problemas, apresentam maior bem-estar psicológico. Já os meninos apresentam melhor bem-estar na medida em que buscam pertença e procuram amigos íntimos e ajuda profissional. A análise do perfil diferencial entre os gêneros em relação às estratégias de coping nos permite identificar uma função discriminante composta por 9 variáveis, que se distribuem diferencialmente de acordo com a utilização de estratégias referentes a papéis de gênero socialmente construídos.

Differential profiling analysis of proteins involved in anti-proliferative effect of interferon-α on renal cell carcinoma cell lines by protein biochip technology

Interferon-alpha (IFN) is widely used for the treatment of progressive renal cell carcinoma (RCC), but its effective response rate is only about 15%. New biomarkers of RCC contributing to the effective IFN treatment are needed to establish a sensitivity test for evaluating if the IFN is effective or not against RCC. All the proteins expressed in the IFN-susceptible and -resistant RCC cell lysates were analyzed by surface enhanced laser desorption ionization (SELDI) mass spectrometry using ProteinChip technology and their different protein expression were detected by the comparison of the profiles between them. We detected the following candidate markers that exhibited peak shifts: a) in the IFN-susceptible cell lines, a candidate marker with a molecular weight of 5,688 Da was detected on a hydrophobic (H4) chip: b) in the IFN-resistant cell lines, candidate markers each with a molecular weight of 8,049, 3,157, 3,993, and 8,959 Da were detected on strong anion exchange (SAX2, pH 9.0, two types) chips, H4 chip, and weak cation exchange (WCX, pH 9.0) chips, respectively. IFN treatment produced no weight increase in these four proteins, and c) candidate marker with a molecular weight of 1,623 Da that was expressed in both cell lines after the IFN treatment was detected on the H4 chip. These data suggest that the ProteinChip system is very useful in identifying proteins showing unique peaks in the RCC cell lines with different IFN susceptibility, and the comparison of these proteins measured in RCC cell lysates may help to identify the IFN sensitivity. Furthermore, the discovery of a susceptibility and or a inhibitory factor may eventually lead to the development of a novel drug targeting the respective factor for the improvement of anticancer chemotherapy.

YW3-10 Differential profiling analysis of proteins involved in anti-proliferative effect of interferon-α on RCC cell lines by protein biochip technology (Young Urologist Research Workshop)

YW3-10 Differential profiling analysis of proteins involved in anti-proliferative effect of interferon-α on RCC cell lines by protein biochip technology (Young Urologist Research Workshop)

Differential profile analysis of neuropsychological functions in schizophrenia

Differential profile analysis of neuropsychological functions in schizophrenia

Differential profile analysis of the General Aptitude Test Battery for rehabilitation clients

Differential profile analysis of the General Aptitude Test Battery for rehabilitation clients