The Local Neuropeptide System of Keratinocytes

Neuropeptides have been known for over 50 years as chemical signals in the brain. However, it is now well established that the synthesis of this class of peptides is not restricted to neurons. For example, human skin not only expresses several functional receptors for neuropeptides but, also, can serve as a local source of neuroactive molecules such as corticotropin-releasing hormone, melanocortins, and β-endorphin. In contrast, an equivalent of the hypothalamic-pituitary axis in the oral mucosa has not been well characterized to date. In view of the differences in the morphology and function of oral mucosal and skin cells, in this review I surveyed the existing evidence for a local synthesis of hypothalamic-pituitary, opiate, neurohypophyseal, and neuroendocrine neuropeptides in both epidermal and oral keratinocytes.

Axonal TDP-43 condensates drive neuromuscular junction disruption through inhibition of local synthesis of nuclear encoded mitochondrial proteins

Mislocalization of the predominantly nuclear RNA/DNA binding protein, TDP-43, occurs in motor neurons of ~95% of amyotrophic lateral sclerosis (ALS) patients, but the contribution of axonal TDP-43 to this neurodegenerative disease is unclear. Here, we show TDP-43 accumulation in intra-muscular nerves from ALS patients and in axons of human iPSC-derived motor neurons of ALS patient, as well as in motor neurons and neuromuscular junctions (NMJs) of a TDP-43 mislocalization mouse model. In axons, TDP-43 is hyper-phosphorylated and promotes G3BP1-positive ribonucleoprotein (RNP) condensate assembly, consequently inhibiting local protein synthesis in distal axons and NMJs. Specifically, the axonal and synaptic levels of nuclear-encoded mitochondrial proteins are reduced. Clearance of axonal TDP-43 or dissociation of G3BP1 condensates restored local translation and resolved TDP-43-derived toxicity in both axons and NMJs. These findings support an axonal gain of function of TDP-43 in ALS, which can be targeted for therapeutic development.

Cortical wiring by synapse-specific control of local protein synthesis

Neurons use local protein synthesis as a mechanism to support their morphological complexity, which requires independent control across multiple subcellular compartments including individual synapses. However, to what extent local translation is differentially regulated at the level of specific synaptic connections remains largely unknown. Here, we identify a signaling pathway that regulates the local synthesis of proteins required for the formation of excitatory synapses on parvalbumin-expressing (PV+) interneurons in the mouse cerebral cortex. This process involves the regulation of the mTORC1 inhibitor Tsc2 by the receptor tyrosine kinase ErbB4, which enables the local control of mRNA translation in a cell type-specific and synapse-specific manner. Ribosome-associated mRNA profiling reveals a molecular program of synaptic proteins that regulates the formation of excitatory inputs on PV+ interneurons downstream of ErbB4 signaling. Our work demonstrates that local protein translation is regulated at the level of specific connections to control synapse formation in the nervous system.

The Inflammatory Feed-Forward Loop Triggered by the Complement Component C3 as a Potential Target in Endometriosis

The complement system is a major component of humoral innate immunity, acting as a first line of defense against microbes via opsonization and lysis of pathogens. However, novel roles of the complement system in inflammatory and immunological processes, including in cancer, are emerging. Endometriosis (EM), a benign disease characterized by ectopic endometrial implants, shows certain unique features of cancer, such as the capacity to invade surrounding tissues, and in severe cases, metastatic properties. A defective immune surveillance against autologous tissue deposited in the peritoneal cavity allows immune escape for endometriotic lesions. There is evidence that the glandular epithelial cells found in endometriotic implants produce and secrete the complement component C3. Here, we show, using immunofluorescence and RT-qPCR, the presence of locally synthesized C3 in the ectopic endometriotic tissue, but not in the eutopic tissue. We generated a murine model of EM via injection of minced uterine tissue from a donor mouse into the peritoneum of recipient mice. The wild type mice showed greater amount of cyst formation in the peritoneum compared to C3 knock-out mice. Peritoneal washings from the wild type mice with EM showed more degranulated mast cells compared to C3 knock-out mice, consistent with higher C3a levels in the peritoneal fluid of EM patients. We provide evidence that C3a participates in an auto-amplifying loop leading to mast cell infiltration and activation, which is pathogenic in EM. Thus, C3 can be considered a marker of EM and its local synthesis can promote the engraftment of the endometriotic cysts.

Testicular Melatonin and Its Pathway in Roe Deer Bucks (Capreolus capreolus) during Pre- and Post-Rut Periods: Correlation with Testicular Involution

Roe deer are seasonal breeders with a complete yearly testicular cycle. The peak in reproductive activity is recorded during summer, the rutting period, with the highest levels of androgens and testicular weight. Melatonin plays a pivotal role in seasonal breeders by stimulating the hypothalamus–pituitary–gonads axis and acting locally; in different species, its synthesis within testes has been reported. The aim of this study was to evaluate the physiological melatonin pattern within roe deer testes by comparing data obtained from animals sampled during pre- and post-rut periods. Melatonin was quantified in testicular parenchyma, along with the genetic expression of enzymes involved in its local synthesis (AANAT and ASMT) and function (UCP1). Melatonin receptors, MT1-2, were quantified both at protein and gene expression levels. Finally, to assess changes in reproductive hormonal profiles, testicular dehydroepiandrosterone (DHEA) was quantified and used for a correlation analysis. Melatonin and AANAT were detected in all samples, without significant differences between pre- and post-rut periods. Despite DHEA levels confirming testicular involution during the post-rut period, no correlations appeared between such involution and melatonin pathways. This study represents the first report regarding melatonin synthesis in roe deer testes, opening the way for future prospective studies in the physiology of this species.

Considerações sobre hormônios e sexo / Considerations on hormones and sex

Os esteroides sexuais, em particular os estrogênios e os androgênios, representam papel fundamental, na modulação da função sexual feminina. As mulheres após a menopausa, devido ao declínio da função ovariana, são dependentes da síntese local de estrogênios nos tecidos alvo extragonadais. Mantendo-se o quadro carêncial ocorre progressivo estreitamento da vagina, e o sintoma genital mais comum é a secura por diminuição da transudação e da quantidade de muco cervical. Esses fenômenos explicam os processos encontrados no período do climatério ou qualquer período em que ocorra a carência estrogênica, característicos da falência hormonal: vaginite atrófica, leucorréia, prurido, dor e sangramento ao coito. Com relação aos androgênios, vários estudos investigaram associações entre androgênios e função sexual em mulheres. Muitos, mas não todos, identificam uma relação entre o desejo sexual e as concentrações séricas de androgênios circulantes. A variabilidade nos resultados provavelmente reflete as limitações em aferir com precisão suas baixas concentrações, o impacto potencial de outros esteroides, as diferenças no desenho do estudo, na seleção das participantes e nos parâmetros de função sexual. Neste artigo, revisamos o conhecimento atual sobre o papel dos estrogênios e androgênios e seu uso clínico em mulheres com declínio da função ovariana.Palavras chave: Hormônios sexuais, Esteroides, Androgênios, Pós-Menopausa, Disfunções sexuais fisiológicasABSTRACTSex steroids, in particular estrogens and androgens, play a fundamental role in the modulation of female sexual function. Postmenopausal women, due to the decline in ovarian function, are dependent on the local synthesis of estrogens in extragonadal target tissues. If the deficiency is maintained, progressive narrowing of the vagina occurs, and the most common genital symptom is dryness due to decreased transudation and the amount of cervical mucus. These phenomena explain the processes found during the climacteric period or any period in which estrogen deficiency occurs, which are characteristic of hormonal failure: atrophic vaginitis, leukorrhea, itching, pain and bleeding on intercourse. With regard to androgens, several studies have investigated associations between androgens and sexual function in women. Many, but not all, identify a relationship between sexual desire and serum concentrations of circulating androgens. The variability in the results probably reflects the limitations in accurately measuring the low concentrations of androgens in women, the potential impact of other steroids, the differences in the study design, in the selection of participants and in the parameters of sexual function. In this article, we review current knowledge about the role of estrogens and androgens and their clinical use in women with declining ovarian function.Keywords: Sex hormones, Steroids, Androgens, Postmenopause, Physiological sexual dysfunctions

Can Extra Adrenal Cortisol Secretion Occur in an Adrenalectomized Patient?

Background: Established medical practice is that adrenalectomized patients require lifelong steroid replacement. We report a case which challenges that assumption. Clinical Case: A 38 year old female underwent transsphenoidal resection of a proven ACTH secreting pituitary adenoma. Post operatively her pituitary function was preserved, but due to residual tumour activity and the desire to preserve fertility, she had a bilateral adrenalectomy one year later and commenced hydrocortisone replacement. Subsequent pituitary imaging did not show recurrence of the tumour and her visual fields were stable, however her ACTH remained elevated at 100 and 130ng/L (09:00 <50 ng/L) despite steroid replacement.14 years after adrenalectomy she noticed weight gain and increased body hair. She also reported sometimes missing her hydrocortisone due to her busy job without ill effects. Her 9 am cortisol pre-hydrocortisone was elevated at 333 nmol/L (9am: 140–690 nmol/L) with ACTH of 203.0 ng/L (09:00 <50ng/L) She was able to suppress cortisol normally after a Dexamethasone suppression test. CT of her adrenals found no adrenal tissue. By now she had weaned herself off hydrocortisone, keeping an emergency supply. SST showed a cortisol of 320 nmol/L at 30 minutes, 360 nmol/L at 60 minutes with an elevated ACTH of 140 ng/L (09:00 <50 ng/L). Her weight gain and body hair growth stabilised. Discussion: We have demonstrated that this adrenalectomized patient has ACTH driven endogenous cortisol secretion which may be due to residual adrenal tissue due to seeding after surgery or the presence of steroid synthesis elsewhere. Extra adrenal sources for glucocorticoid production are known such as skin, gonads and thymus. However, the levels are insufficient to mount a significant stress response. There is evidence of adrenal regeneration in adrenalectomized animals. The regeneration is primarily of the adrenal cortex and does not involve the medulla. There has been one case report in literature of a 11 year old German boy who had adrenal regeneration detected on adrenal scintigraphy (Bilateral normal adrenal glands with normal activity) 13 years after adrenalectomy for Cushing’s disease. References: Gotlieb N, Albaz E, Shaashua L, et al. Regeneration of Functional Adrenal Tissue Following Bilateral Adrenalectomy. Endocrinology. 2018;159(1):248–259. doi:10.1210/en.2017-00505 Taves MD, Gomez-Sanchez CE, Soma KK. Extra-adrenal glucocorticoids and mineralocorticoids: evidence for local synthesis, regulation, and function. Am J Physiol Endocrinol Metab. 2011;301(1):E11-E24. doi:10.1152/ajpendo.00100.2011

Evaluation of Free Light Chains of Immunoglobulins in Seminal Plasma of Infertile Patients: Preliminary Data

Seminal plasma is a complex fluid with various components (proteins, enzymes, macro-and microelements, lipids and nutrients) and its role is fundamental for spermatozoa motility, viability and fertilizing capacity maintenance. Many molecules have been measured in seminal plasma to explore some secretion functions of male accessory glands, but effects of biochemical components in human seminal plasma are still debated. Immunoglobulin-free light chains (FLCs) κ and λ are produced by plasma cells in slight excess for the need of immune response and are therefore assayable in blood and in other biological fluids, such as urine, saliva, liquor and synovial fluid. Recently, different biological functions have been attributed to these molecules, suggesting that they are not just a secondary product of immunoglobulin synthesis. No data are reported about presence of FLCs in seminal plasma and their role in physiology of male reproductive system and/or in pathophysiology of infertility. The aims of our study were to investigate the presence and detectability of FLCs in seminal plasma and to evaluate the usefulness of this assay in the diagnostic approach to infertility patients. We enrolled 32 patients aged 19-40 ys, affected by primary infertility; among them, 7 were normospermic (mean±SEM concentration 100.0±16.0 *106/ml; progressive motile forms 39.1±4.9%; normal forms 45.3±4.5%), 25 were oligo- and/or asthenoteratospermic (mean±SEM concentration 23.8±5.4*106/ml; progressive motile forms 19.3±4.1%; normal forms 36.05±2.7%); moreover, 17 patients presented II-IV degree varicocele (VAR) according to Dubin-Amelar classification by Doppler technique, the remaining 15 patients did not present varicocele (NO-VAR). After abstinence for 3-5 days, semen samples were collected. FLCs concentrations were assayed by turbidimetric method. Standard semen analysis was performed according to WHO laboratory manual for the examination and processing of human semen, fifth edition, 2010. As main results, independently from sperm count, a significantly difference was observed concerning FLCs concentrations, with higher levels of k and k/λ ratio in NO-VAR vs VAR patients (mean±SEM k 36.4±13.2 vs 17.7±9.0 g/l and 7.7±2.9 vs 2.65±0.7, respectively; p<0.05). λ FLCs did not significantly differ between the two groups. This work shows for the first time that FLCs are assayable in seminal plasma, even if their source is to be determined (plasma filtration or local synthesis from lymphoid tissue in accessory gland). Our preliminary data also showed a peculiar pattern with prevalence of k FLCs in infertile patients without VAR, suggesting that FLCs could be in interesting field of investigation in idiopathic infertility. Further studies in large and stratified patients may reveal a possible usefulness of FLCs as a biological marker and/or gain insight about their etiopathogenetic role in male infertility.

Indolaminergic System in Adult Rat Testes: Evidence for a Local Serotonin System

Serotonin (5-HT) is member of a family of indolamine molecules that participate in a wide variety of biological processes. Despite its important role in the regulation of local blood systems, little is known about the physiological function of 5-HT in reproductive organs, its functional implications, and its role in the reproduction of mammals. In the present work, we evaluated the localization and distribution of 5-HT (using histochemical analysis of indolamines) and different components of the serotoninergic system in rat testes. We detected local synthesis and degradation through immunofluorescence and western blot analyses against the TPH1, MAOA, 5-HTT, and VMAT1 serotonin transporters. We also identified the localization and distribution of the 5-HT1B, 5-HT2A, and 5-HT3A receptors. RT-PCR results showed the presence of the Tph1, Maoa, Slc6a4, and Htr3a genes in testes and in the brain stem (Tph1 was used as a negative control). High-performance liquid chromatography was used to determine the presence of 5-HT and the activity of tryptophan hydroxylase in testes homogenates in vitro. Our observations suggest that TPH1 activity and local 5-HT synthesis befall in rat testes. We propose that 5-HT could participate in the regulation of testosterone synthesis and in the spermatogenesis process via local serotoninergic system. However, more studies are needed before concluding that rat testes, or those of other mammals, contain an active form of tryptophan hydroxylase and produce 5-HT.

Management Of Enterprise Development Strategies Formation: Analysis And Synthesis Methods

The purpose of the presented research is to analyze, systematize and formulate an enterprise strategy based on the analysis and synthesis of opportunities and threats of the external environment in combination with its strengths and weaknesses. Unconventional structuring of SWOT-factors, as well as their local synthesis for determination of the driving and inhibiting forces of the enterprise, has been offered. Ways to identify trends and patterns of external and internal environment based on a generalized model of SWOT-factors have been identified. Decisive in formulating a strategy (self-organized in nature) is a harmonization model of the goals system. General theoretical methods (analogy, abstraction, analysis and synthesis, idealization, imaginary modeling) and practical methods of research (observation, comparison, matching, analysis of statistical and other indicators) were used to solve the problems of the research. Methods such as empirical, statistical, graphical, etc. were used to process and summarize information.Thus, the approach study to the enterprise strategy formation based on the analysis and synthesis of the environment opportunities and threats in combination with its strengths and weaknesses allows to state that the development of a common structured list of major areas of enterprise activity is carried out taking into account the synthesis of all SWOT-analysis of the brand product, competitors and market segments, which allows the company to achieve a certain specific purpose of its activities.