Disease-Specific haptoglobin N-Glycosylation in inflammatory disorders between cancers and benign diseases of 3 types of female internal genital organs.

BACKGROUND: N-glycosylation of the haptoglobin is closely related to pathological states. This study aims to evaluate the association of glycosylation of disease-specific Hp (DSHp) ß chain with different pathological states of the cervix, uterus, and ovary to explore differences in their inflammatory responses and to screen potential biomarkers to distinguish cancer from benign diseases. METHODS: DSHp-ß chains of 1956 patients with cancers and benign diseases located in the cervix, uterus, and ovary organs were separated from serum immunoinflammatory-related protein complexes (IIRPCs). The N-glycopeptides from DSHp-ß chains were detected using mass spectrometry, followed by an analysis of machine learning algorithms. RESULTS: 55 N-glycopeptides at N207/N211, 19 at N241, and 21 at N184 glycosylation sites of DSHp for each sample were identified. Fucosylation and sialylation of DSHp in cervix, uterus, and ovary cancer were significantly increased compared to their corresponding benign diseases (p < 0.001). The cervix diagnostic model, a combination of G2N3F, G4NFS, G7N2F2S5, GS-N&GS-N, G2N2&G4N3FS, G7N2F2S5, G2S2&G-N, and GN2F&G2F at N207/N211 sites, G3NFS2 and G3NFS at N241site, G9N2S, G6N3F6, G4N3F5S, G4N3F4S2, and G6N3F4S at N184 site), has shown a good diagnostic capability to distinguish cancer from benign diseases, with the area under curve (AUC) of 0.912. The uterus diagnostic model including G4NFS, G2S2&G2S2, G3N2S2, GG5N2F5, G2&G3NFS, and G5N2F3S3 at N207/N211 sites, and G2NF3S2 at N184 site, with an AUC of 0.731. The ovary diagnostic model including G2N3F, GF2S-N &G2F3S2, G2S&G2, and G2S&G3NS at N207/N211 sites; G2S and G3NFS at N241 site, G6N3F4S at N184 site, with an AUC of 0.747. CONCLUSIONS: These findings provide insights into differences in organ-specific inflammatory responses of DSHp for different pathological states among the organs of the cervix, uterus, and ovary.

The Effects of Clove and Its Constituents on Reproductive System: a Comprehensive Review.

Clove with the scientific name of Syzygium aromaticum (L.) Merr. & L.M. Perry is an evergreen tree in which its buds are used for medicinal purposes. Traditional medicine manuscripts as well as recent studies reported its effects on male and female reproductive systems. The aim of this study is to investigate the reported contradictory effects of clove and its phytochemicals on the reproductive system of both males and females. All types of in vitro, animal, and human studies of clove and its main constituents in the field of reproductive systems were collected via searching electronic databases including PubMed and Scopus from the onset till 2021. In this review, 76 articles were included, of which 25 were related to male reproduction, 32 were related to female reproduction, and 19 were related to reproductive malignancies. Analysis of the literature indicates the effects of clove and its constituents especially eugenol and ß-caryophyllene on the level of sex hormones, fertility, sperm abnormalities, endometriosis, menstrual cycle, as well as gynecological infections, and reproductive tumors. The main mechanism of clove has not been understood yet but it seems that different parameters affect its pharmacological activity including the type of extract, dose, and duration of administration as well as the primary cause of the disorder. According to the effects of clove on different parts of the reproductive system, it seems that it can be a suitable candidate for related disorders, provided that more and more detailed studies are done on it.

Evaluation of the Concentration of Selected Elements in Patients with Cancer of the Reproductive Organs with Respect to Treatment Stage-Preliminary Study.

(1) Background: The aim of this study was to assess the concentrations of selected elements in female patients with cancer of the reproductive organs, taking into account the stage of treatment. (2) Methods: The study sample consisted of 51 patients with advanced endometrial cancer and ovarian cancer, undergoing chemotherapy. The median age of the studied patients with endometrial cancer was 66.0 years (IQR: from 60.75 to 70.25), and with ovarian cancer­60.0 years (IQR: from 49.0 to 64.0). Each of the qualified women, after consent to participate in the study, had her blood drawn several times (before surgery, the first course of chemotherapy, the third course of chemotherapy, and the sixth course of chemotherapy) in order to determine serum levels of macro- and micronutrients (Na, Mg, Ca, Zn, P, Cu, Fe, Cd, Ni, and Sr). (3) Results: In the study group of patients with cancer of the reproductive tract, the concentrations of iron (<0.001), magnesium (0.038), sodium (0.014), and nickel (0.037) varied significantly over the course of the study. The analysis showed that the interaction between the stage of chemotherapy and the type of cancer had an effect on the concentrations of magnesium and cadmium (p < 0.05). (4) Conclusions: In the studied group of patients with ovarian and endometrial cancer, the applied chemotherapy significantly changed the concentrations of Fe, Na, and Ni, regardless of the type of tumor. Changes in Mg and Cd concentrations resulted from the interaction between the stage of chemotherapy and the type of cancer. The results of serum concentrations of selected elements in women with cancer of the reproductive organs may help understand the physiological changes resulting from the applied chemotherapy.

Effects of Polychlorinated Biphenyls on Animal Reproductive Systems and Epigenetic Modifications.

Polychlorinated biphenyls (PCBs) are a group of highly toxic endocrine-disrupting chemicals comprising 209 homologs. PCBs are extensively found in the environment and can induce typical estrogenic and profound, long-lasting effects on animals. In this article, the introduction of PCB residues into the environment and the pathways of PCB enrichment in animals are described. PCBs are widely deposited and eventually accumulate in human tissues and body fluids through biomagnification. PCBs can significantly decrease animal fertility and interfere with endocrine processes, leading to the development of various diseases and even cancer. The effects of PCBs on the reproductive systems of animals can also be passed to their offspring, indicating that PCBs may affect the epigenetic modification process. There is currently no treatment to effectively inhibit the toxicity of PCBs in organisms; therefore, the severity of PCB toxicity needs to be widely recognized.

Transition of Radioactive Cesium Deposition in Reproductive Organs of Free-Roaming Cats in Namie Town, Fukushima.

We investigated the internal contamination by radioactive cesium associated with the FDNPP accident, in the testes or uterus and ovaries of free-roaming cats (Felis silvestris catus), which were protected by volunteers in the Namie Town, Fukushima. A total of 253 samples (145 testes and 108 uterus and ovaries) obtained from adult cats and 15 fetuses from 3 pregnant female cats were measured. Free-roaming cats in Namie Town had a higher level of radioactive contamination in comparison to the control group in Tokyo, as the 134Cs + 137Cs activity concentration ranged from not detectable to 37,882 Bq kg-1 in adult cats. Furthermore, the radioactivity in the fetuses was almost comparable to those in their mother's uterus and ovaries. The radioactivity was also different between several cats protected in the same location, and there was no significant correlation with ambient dose-rates and activity concentrations in soil. Moreover, radioactive cesium levels in cats decreased with each year. Therefore, it is likely that decontamination work in Namie Town and its surroundings could affect radioactive cesium accumulation, and thus possibly reduce the internal radiation exposure of wildlife living in contaminated areas. It is hence necessary to continue radioactivity monitoring efforts for the residents living in Namie Town.

Microplastic pollution as a grand challenge in marine research: A closer look at their adverse impacts on the immune and reproductive systems.

Microplastic (MP) pollution of the marine environment is now a growing global concern posing a threat to a variety of species through the ingestion and transfer within food webs. This is considered a potential toxicological threat to marine species due to the chemical additives used to make many plastic products, or the persistent organic pollutants that may accumulate on them while residing in the environment. While the presence of MPs in the marine environment is widely documented, there are no other review articles providing a summary of published effect studies of MPs on the immune and reproductive systems of marine species. This manuscript reviews reproductive and immune-system changes in response to MPs in 7 and 9 species, respectively. Some species such as Mytilus galloprovincialis and oyster Crassostrea gigas were investigated in multiple papers. Most studies have been conducted on invertebrates, and only 3 studies have been performed on vertebrates, with exposure times ranging between 30 min and 60 days. A review of the literature revealed that the most common MPs types studied in relation to adverse impacts on immune system and reproductive success in marine species were polystyrene (PS) and polyethylene (PE). The immune system's responses to MPs exposure varied depending on the species, with altered organismal defense mechanisms and neutrophil function observed in fish and changes in lysosomal membrane stability and apoptotic-like nuclear alterations in phagocytes reported in invertebrate species. Reproductive responses to MPs exposure, varied depending on species, but included significant reduction in gamete and oocyte quality, fecundity, sperm swimming speed, and quality of offspring. The lack of published data means that developing a clear understanding of the impact across taxonomic groups with different feeding and behavioral traits is often difficult. Further work is required to better understand the risk MPs pose to the immune and reproductive systems of marine species in order to fully evaluate the impact these ubiquitous pollutants are having on marine ecosystems and the associated goods and services they provide.

Trichome Density in Relation to Volatiles Emission and 1,8-Cineole Synthase Gene Expression in Thymus albicans Vegetative and Reproductive Organs.

1,8-Cineole is the main volatile produced by Thymus albicans Hoffmanns. & Link 1,8-cineole chemotype. To understand the contribution of distinct plant organs to the high 1,8-cineole production, trichome morphology and density, as well as emitted volatiles and transcriptional expression of the 1,8-cineole synthase (CIN) gene were determined separately for T. albicans leaves, bracts, calyx, corolla and inflorescences. Scanning electron microscopy (SEM) and stereoscope microscopy observations showed the highest peltate trichome density in leaves and bracts, significantly distinct from calyx and corolla. T. albicans volatiles were collected by solid phase micro extraction (SPME) and analyzed by gas chromatography-mass spectrometry (GC/MS) and by GC for component identification and quantification, respectively. Of the 23 components identified, 1,8-cineole was the dominant volatile (57-93 %) in all T. albicans plant organs. The relative amounts of emitted volatiles clearly separated vegetative from reproductive organs. Gene expression of CIN was assigned to all organs analyzed and was consistent with the relatively high emission of 1,8-cineole in leaves and bracts. Further studies will be required to analyze monoterpenoid biosynthesis by each type of glandular trichome.

Gender proteomics II. Which proteins in sexual organs.

In continuity with the review dealing with differences by gender in non-sexual organs [1], this review collects data on the proteomes of the sexual organs as involved in human reproduction, under both physiological and pathological conditions. It also collects data on the tissue structures and biological fluids typical of pregnancy, such as placenta and amniotic fluid, as well as what may be tested on preimplantation embryos during medically assisted reproduction. The review includes as well mention to all fluids and secretions connected with sex organs and/or reproduction, including sperm and milk, to exemplify two distinctive items in male and female physiology. SIGNIFICANCE: The causes of infertility are only incompletely understood; the same holds for the causes, and even the early markers, of the most frequent complications of pregnancy. To these established medical challenges, present day practice adds new issues connected with medically assisted reproduction. Omics approaches, including proteomics, are building the database for basic knowledge to possibly translate into clinical testing and eventually into medical routine in this critical branch of health care.

Estimating mating rates in wild Drosophila melanogaster females by decay rates of male reproductive proteins in their reproductive tracts.

Female Drosophila melanogaster frequently mate with multiple males in nature as shown through parentage analysis. Although polyandry is well documented, we know little about the timing between mating events in wild Drosophila populations due to the challenge of following behaviours of individual females. In this study, we used the presence of a male reproductive protein that is transferred to the female during mating (Sex Peptide, SP) to determine whether she had recently mated. We sampled females throughout the day, conducted control matings to determine the decay rate of SP within the female reproductive tract and performed computer simulations to fit the observed proportion of mated females to a nonhomogenous Poisson process that defined the expected time between successive matings for a given female. In our control matings, 100% of mated females tested positive for SP 0.5 h after the start of mating (ASM), but only 24% tested positive 24 h ASM. Overall, 35% of wild-caught females tested positive for the presence of SP. Fitting our observed data to our simple nonhomogenous Poisson model provided the inference that females are mating, on average, approximately every 27 h (with 95% credibility interval 23-31 h). Thus, it appears that females are mating a bit less frequently that once per day in this natural population and that mating events tend to occur either early in the morning or late in the afternoon.

Intracellular Tenofovir and Emtricitabine Anabolites in Genital, Rectal, and Blood Compartments from First Dose to Steady State.

The pharmacokinetics (PK) of tenofovir-diphosphate (TFV-DP) and emtricitabine-triphosphate (FTC-TP), the active anabolites of tenofovir disoproxil fumarate (TDF), and emtricitabine (FTC) in blood, genital, and rectal compartments was determined in HIV-positive and seronegative adults who undertook a 60-day intensive PK study of daily TDF/FTC (plus efavirenz in HIV positives). Lymphocyte cell sorting, genital, and rectal sampling occurred once per subject, at staggered visits. Among 19 HIV-positive (3 female) and 21 seronegative (10 female) adults, TFV-DP in peripheral blood mononuclear cells (PBMC) accumulated 8.6-fold [95% confidence interval (CI): 7.2-10] from first-dose to steady-state concentration (Css) versus 1.7-fold (95% CI: 1.5-1.9) for FTC-TP. Css was reached in ∼11 and 3 days, respectively. Css values were similar between HIV-negative and HIV-positive individuals. Css TFV-DP in rectal mononuclear cells (1,450 fmol/106 cells, 898-2,340) was achieved in 5 days and was >10 times higher than PBMC (95 fmol/106 cells, 85-106), seminal cells (22 fmol/106 cells, 6-79), and cervical cells (111 fmol/106 cells, 64-194). FTC-TP Css was highest in PBMC (5.7 pmol/106 cells, 5.2-6.1) and cervical cells (7 pmol/106 cells, 2-19) versus rectal (0.8 pmol/106 cells, 0.6-1.1) and seminal cells (0.3 pmol/106 cells, 0.2-0.5). Genital drug concentrations on days 1-7 overlapped with estimated Css, but accumulation characteristics were based on limited data. TFV-DP and FTC-TP in cell sorted samples were highest and achieved most rapidly in CD14+ compared with CD4+, CD8+, and CD19+ cells. Together, these findings demonstrate cell-type and tissue-dependent cellular pharmacology, preferential accumulation of TFV-DP in rectal mononuclear cells, and rapid distribution into rectal and genital compartments.

A Proteomic Analysis of the Body Wall, Digestive Tract, and Reproductive Tract of Brugia malayi.

Filarial worms are parasitic nematodes that cause devastating diseases such as lymphatic filariasis (LF) and onchocerciasis. Filariae are nematodes with complex anatomy including fully developed digestive tracts and reproductive organs. To better understand the basic biology of filarial parasites and to provide insights into drug targets and vaccine design, we conducted a proteomic analysis of different anatomic fractions of Brugia malayi, a causative agent of LF. Approximately 500 adult female B. malayi worms were dissected, and three anatomical fractions (body wall, digestive tract, and reproductive tract) were obtained. Proteins from each anatomical fraction were extracted, desalted, trypsinized, and analyzed by microcapillary reverse-phase liquid chromatography-tandem-mass spectrometry. In total, we identified 4,785 B. malayi proteins. While 1,894 were identified in all three anatomic fractions, 396 were positively identified only within the digestive tract, 114 only within the body wall, and 1,011 only within the reproductive tract. Gene set enrichment analysis revealed a bias for transporters to be present within the digestive tract, suggesting that the intestine of adult filariae is functional and important for nutrient uptake or waste removal. As expected, the body wall exhibited increased frequencies of cytoskeletal proteins, and the reproductive tract had increased frequencies of proteins involved in nuclear regulation and transcription. In assessing for possible vaccine candidates, we focused on proteins sequestered within the digestive tract, as these could possibly represent "hidden antigens" with low risk of prior allergic sensitization. We identified 106 proteins that are enriched in the digestive tract and are predicted to localize to the surface of cells in the the digestive tract. It is possible that some of these proteins are on the luminal surface and may be accessible by antibodies ingested by the worm. A subset of 27 of these proteins appear especially promising vaccine candidates as they contain significant non-cytoplasmic domains, only 1-2 transmembrane domains, and a high degree of homology to W. bancrofti and/or O. volvulus.

Expression of steroid hormone receptors in the genital structures of a true hermaphrodite pug dog.

Hermaphroditism is a rare and a not well-understood disordered sexual development (DSD) in dogs. The objective of the study was to analyse the sex steroid hormone receptor (STHR) expression patterns in the internal genital structures, because the responsiveness of the different tissue types to the steroid hormones may have a key role in pathological alterations based on DSDs. Furthermore, the adhesion molecule ß-catenin was investigated by means of immunohistochemistry because of its important role in development, tissue integrity and disease. Molecular sexing was performed via PCR targeting DBX/DBY genes to identify the pug dog as a true XX hermaphrodite. The portions of uterine tissue revealed comparable expression patterns for STHRs as investigated in normal female reproductive tissue. In the male parts, ß-catenin showed strong expression in the Sertoli cells of the seminiferous tubules; this was in contrast to normal testicular tissue. Likewise, the layers of smooth muscle actin-positive cells surrounding the seminiferous tubules were reduced in the hermaphrodite. The results of this study deepen the knowledge of tissue characteristics in a hermaphrodite dog and highlight the importance of early diagnosis because the STH responsiveness in maldeveloped reproductive tissue might lead to serious problems for the dog.

Regulatory neuropeptides (ghrelin, obestatin and nesfatin-1) levels in serum and reproductive tissues of female and male rats with fructose-induced metabolic syndrome.

Although, the exact mechanisms underlying the development of the metabolic syndrome (MetS) are not still completely understood, obesity, circulated peptide hormone levels and their interaction with genetic factors are considered largely responsible. The purpose of this study is to explore how the levels of ghrelin, obestatin (OBS) and NUCB2/nesfatin-1 (NES)/NUCB2 change in serum and the reproductive tissues of female and male rats with fructose-induced metabolic syndrome, and whether the levels of each hormone is correlated with the hormones involved with fertility. Experiments were conducted on 5-week-old Sprague-Dawley male and female rats assigned to either a control group or a MetS group. Controls were fed standard rat food and water ad libitum, while the MetS group was fed standard food with 10% (v/v) fructose solution added to their drinking water for 12 weeks with a 12/12h photoperiod circle. Then, all animals were sacrificed after a one night fast. Peptides levels in the serum and reproductive tissues of rats were studied using the ELISA method while the immunoreactivity of reproductive system peptide hormones were shown by immunohistochemical staining method. Furthermore, the other biochemical parameters were measured using Konelab-60 equipment and infertility hormones were measured with Immulite2000. Fasting serum insulin, glucose, triglyceride, alanine aminotransferase (ALT), gamma glutamyl transpeptidase (GGT), low-density lipoprotein cholesterol (LDL-C), and total cholesterol (TC) levels were statistically significantly higher, and the amount of high density lipoprotein cholesterol (HDL-C) was significantly lower, in the MetS groups. Serum and tissue supernatant NES levels were significantly higher in the rats with MetS than the control group. Ghrelin, OBS and NES were expressed in the cytoplasm, concentrated around the apical parts of the epithelial cells in the reproductive tissues of the rats. The amounts of ghrelin were lower in the reproductive tissues of the animals with MetS, while NES levels in the same tissues increased. Obestatin also decreased, though not in the seminal glands.

Novel parasitic nematode-specific protein of Setaria digitata largely localized in longitudinal muscles, reproductive systems and developing embryos.

Parasitic nematodes may have common properties in parasitizing the host which are conferred by related parasitic proteins encoded by their genome. A novel protein characterized from bovine filarial nematode Setaria digitata was found to be present only in the parasitic nematodes and expressed at all the stages of the nematode's life. In immunohistochemical staining using polyclonal antibodies prepared against recombinant S. digitata protein, the highest expression of S. digitata novel protein (SDNP) was seen in the longitudinal muscles of the body wall of adult males and females indicating its possible involvement in parasite locomotion. Moderate expression was observed in the reproductive organs of both sexes while showing gradual increase in the expression as the development of the reproductive tissue progressed suggesting its role in tissue transformation in male and female reproduction. A low level of expression was observed in the cuticle, syncytial hypodermis region, lateral line and the intestinal wall. Further, the expression of SDNP was also seen in developing microfilaria within the uterus of female worms, developing spermatozoa of males and different developmental stages of embryos implicating its involvement in nematode growth and development. Subcellular localization of SDNP carried out in yeast, Pichia pastoris using green fluorescence construct revealed that this protein localized mainly in the nucleus and partly in the cytoplasm. Comprehensive bioinformatics analyses indicated that this protein contains a nuclear localization signal, RNAP_Rpb7_N_like domain, regions that are homologous to a part of the nuclear factor localization-like domain, interdomain linkers of muscle specific twitchin kinase of Caenorhabditis elegans and calcium-dependent protein kinase isoform CDPK1 of Arabidopsis thaliana. Therefore, considering all the outcomes together, it can be suggested that the SDNP is a parasitic nematode-specific, nuclear and cytoplasmic protein that is likely to be regulated by reversible phosphorylation-dephosphorylation reaction, expressed in all the stages of nematode's life having pivotal functional roles in muscle, reproductive systems, embryogenesis, and also in the growth and development.

Serotonin signaling in the brain of adult female mice is required for sexual preference.

A role for serotonin in male sexual preference was recently uncovered by our finding that male mutant mice lacking serotonin have lost sexual preference. Here we show that female mouse mutants lacking either central serotonergic neurons or serotonin prefer female over male genital odors when given a choice, and displayed increased female-female mounting when presented either with a choice of a male and a female target or only with a female target. Pharmacological manipulations and genetic rescue experiments showed that serotonin is required in adults. Behavioral changes caused by deficient serotonergic signaling were not due to changes in plasma concentrations of sex hormones. We demonstrate that a genetic manipulation reverses sexual preference without involving sex hormones. Our results indicate that serotonin controls sexual preference.

Pharmacokinetics and pharmacodynamics in HIV prevention; current status and future directions: a summary of the DAIDS and BMGF sponsored think tank on pharmacokinetics (PK)/pharmacodynamics (PD) in HIV prevention.

Thirty years after its beginning, the HIV/AIDS epidemic is still raging around the world. According to UNAIDS, in 2011 alone 1.7M deaths were attributable to AIDS, and 2.5M people were newly infected by the virus. Despite the success in treating HIV-infected people with potent antiretroviral drugs, preventing HIV infection is the key to ending the epidemic. Recently, the efficacy of topical and systemic antiviral chemoprophylaxis (i.e., preexposure prophylaxis or "PrEP"), using the same drugs used for HIV treatment, has been demonstrated in a number of clinical trials. However, results from other trials have been inconsistent, especially those evaluating PrEP in women. These inconsistencies may result from our incomplete understanding of pharmacokinetics (PK)/pharmacodynamics (PD) at the mucosal sites of sexual transmission: the male and female gastrointestinal and reproductive tracts. The drug concentrations used in these trials were derived from those used for treatment; however, we still do not know the relationship between the therapeutic and the preventive dose. This article presents the first comprehensive review of the available data in the HIV pharmacology field from animal models to human studies, and outlines gaps, challenges, and future directions. Addressing these pharmacological gaps and challenges will be critical in selecting and advancing future PrEP candidates and strategies with the greatest impact on the HIV epidemic.

Temporally variable selection on proteolysis-related reproductive tract proteins in Drosophila.

In order to gain further insight into the processes underlying rapid reproductive protein evolution, we have conducted a population genetic survey of 44 reproductive tract-expressed proteases, protease inhibitors, and targets of proteolysis in Drosophila melanogaster and Drosophila simulans. Our findings suggest that positive selection on this group of genes is temporally heterogeneous, with different patterns of selection inferred using tests sensitive at different time scales. Such variation in the strength and targets of selection through time may be expected under models of sexual conflict and/or host-pathogen interaction. Moreover, available functional information concerning the genes that show evidence of selection suggests that both sexual selection and immune processes have been important in the evolutionary history of this group of molecules.

Haematology, genotoxicity, enzymatic activity and histopathology as biomarkers of metal pollution in the shrew Crocidura russula.

Haematological (WBC, RBC, Hgb and Hct) and genotoxicity (MNT) parameters, hepatic enzymatic activities (GST, GPx and GR), and a histopathological evaluation of liver, kidneys and gonads were assessed as general biomarkers of metal pollution in the shrew Crocidura russula inhabiting a pyrite mining area. Specimens exposed to metals presented a few significant alterations when compared with reference animals: GST activity decreased; micronuclei increased; and evident liver alterations related to metal exposure were observed. On the basis of all the parameters studied, age was an important factor that partly explained the observed variation, whereas sex was the least important factor. Significant correlations were also found between heavy metal concentrations and biomarkers evaluated, demonstrating the great influence of these metals in the metabolic alterations. To the best of our knowledge, these data constitute the first measurements of a battery of biomarkers in shrews from a mine site and are among the few available for insectivorous mammals.

Lipopolysaccharide modulation of thyrotropin-releasing hormone (TRH) and TRH-like peptide levels in rat brain and endocrine organs.

Lipopolysaccharide (LPS) is a proinflammatory and depressogenic agent whereas thyrotropin-releasing hormone (TRH; pGlu-His-Pro-NH2) is an endogenous antidepressant and neuroprotective peptide. LPS and TRH also have opposing effects on K+ channel conductivity. We hypothesized that LPS can modulate the expression and release of not only TRH but also TRH-like peptides with the general structure pGlu-X-Pro-NH2, where "X" can be any amino acid residue. The response might be "homeostatic," that is, LPS might increase TRH and TRH-like peptide release, thereby moderating the cell damaging effects of this bacterial cell wall constituent. On the other hand, LPS might impair the synthesis and release of these neuropeptides, thus facilitating the induction of early response genes, cytokines, and other downstream biochemical changes that contribute to the "sickness syndrome." Sprague-Dawley rats (300 g) received a single intraperitoneal injection of 100 microg/kg LPS. Animals were then decapitated 0, 2, 4, 8, and 24 h later. Serum cytokines and corticosterone peaked 2 h after intraperitoneal LPS along with a transient decrease in serum T3. TRH and TRH-like peptides were measured by a combination of high-performance liquid chromatography and radioimmunoassay. TRH declined in the nucleus accumbens and amygdala in a manner consistent with LPS-accelerated release and degradation. Various TRH-like peptide levels increased at 2 h in the anterior cingulate, hippocampus, striatum, entorhinal cortex, posterior cingulate, and cerebellum, indicating decreased release and clearance of these peptides. These brain regions are part of a neuroimmunomodulatory system that coordinates the behavioral, endocrine, and immune responses to the stresses of sickness, injury, and danger. A sustained rise in TRH levels in pancreatic beta-cells accompanied LPS-impaired insulin secretion. TRH and Leu-TRH in prostate and TRH in epididymis remained elevated 2-24 h after intraperitoneal LPS. We conclude that these endogenous neuroprotective and antidepressant-like peptides both mediate and moderate some of the behavioral and toxic effects of LPS.

Assessing the potential of immunohistochemistry for systematic gene expression profiling.

Immunohistochemistry (IHC) is a powerful technique for identifying sites of protein expression in tissues at the cellular and sub-cellular level. Here we have investigated the potential of using IHC for genome-wide expression screening by measuring the success rate and specificity of a panel of 35 monoclonal antibodies recognizing 5 well characterised CD antigens. Antibodies were pre-screened on acetone fixed frozen sections of spleen, tonsil and colon tissues. 19/35 antibodies gave staining with a success rate of 0/7 for JAM-2, 1/4 for CD99, 3/6 for CD138, 5/8 for CD45 and 10/10 for MHC-class II. 16/19 of these antibodies also gave staining on formalin fixed paraffin embedded tissue sections of tonsil and colon. All antibodies that had given staining were then profiled on tissues presented in human tissue microarrays. In the frozen microarrays 216 cores from 29 normal tissue types were present and in the formalin fixed paraffin array 344 cores from 35 normal and 4 cancers were represented. Where multiple antibodies were positive, there was evidence of consistent staining of the same tissues with several antibodies. In some cases differences in staining were observed potentially due to differential splice variants, polymorphisms or protein modification. With some antibodies there was evidence of cross-reactivity to inappropriate cells or structures. In addition the staining intensity with formalin fixation was changed quantitatively for some antibodies and in a few cases qualitatively, representing differential sensitivity of specific and non-specific epitopes to fixation. Accordingly, whilst IHC has potential for describing protein expression of unknown genes, these results emphasise a need to systematically address issues of specificity and sensitivity if appropriate profiles are to be described.