Somatic Organs Research Articles

Extended longevity of termite kings and queens is accompanied by extranuclear localization of telomerase in somatic organs and caste-specific expression of its isoforms.

Kings and queens of termites are endowed with an extraordinary longevity coupled with lifelong fecundity. We recently reported that termite kings and queens display a dramatically increased enzymatic activity and abundance of telomerase in their somatic organs when compared to short-lived workers and soldiers. We hypothesized that this telomerase activation may represent a noncanonical pro-longevity function, independent of its canonical role in telomere maintenance. Here, we explore this avenue and investigate whether the presumed noncanonical role of telomerase may be due to alternative splicing of the catalytic telomerase subunit TERT and whether the subcellular localization of TERT isoforms differs among organs and castes in the termite Prorhinotermes simplex. We empirically confirm the expression of four in silico predicted splice variants (psTERT1-A, psTERT1-B, psTERT2-A, psTERT2-B), defined by N-terminal splicing implicating differential localizations, and C-terminal splicing giving rise to full-length and truncated isoforms. We show that the transcript proportions of the psTERT are caste- and tissue-specific and that the extranuclear full-length isoform TERT1-A is relatively enriched in the soma of neotenic kings and queens compared to their gonads and to the soma of workers. We also show that extranuclear TERT protein quantities are significantly higher in the soma of kings and queens compared to workers, namely due to the cytosolic TERT. Independently, we confirm by microscopy the extranuclear TERT localization in somatic organs. We conclude that the presumed pleiotropic action of telomerase combining the canonical nuclear role in telomere maintenance with extranuclear functions is driven by complex TERT splicing.

OR04-05 Prenatal Phthalate Exposure Alters The Pattern Of Fetal And Neonatal Endocrine Puberty In A Sex-specific Manner

Abstract Disclosure: M.A. Bunnell: None. P. Lin: None. S. Zhou: None. C. Park: None. S. Wang: None. Q. Hu: None. B. Marzbanabbasabadi: None. J. Oh: None. B. Wang: None. M. Laws: None. E. Fletcher: None. Z. Liu: None. E. Joo: None. H. Qiao: None. J. Flaws: None. C. Ko: None. Endocrine puberties (EP) are sex-specific transient elevations of gonadotropins and testosterone (T) in males and estradiol (E2) in females. Fetal EP occurs mid-gestation, neonatal EP is stimulated by the clearance of maternal estrogen after birth, and finally, reproductive function arises during classical puberty. EPs are critical for normal development and the sexual dimorphism of reproductive and some somatic organs. Disrupted T or E2 levels during fetal or neonatal EP lead to reproductive abnormalities in adults. The endocrine disrupting effects of phthalates have been well-documented in humans and animal models. Prepubescent children are highly susceptible to phthalate toxicity. In utero, exposure to phthalates causes gender-specific reproductive abnormalities. In this study, we tested the working hypothesis that in utero exposure to phthalates alters T and E2 levels in fetal and newborn mice in a gender-specific manner. Pregnant CD-1 dams (n=5-6/treatment/time point) were orally administered an environmentally relevant phthalate mixture (20 μg/Kg/day) or vehicle daily from embryonic day (ED) 10.5 until birth. On ED16.5, and postnatal hours (h) 1, 3, and 6, fetal tissues and neonatal sera and tissues were sampled for hormonal assays. Fetal and neonatal gonads were collected for testicular T and ovarian E2 measurements. The amount of serum per embryo was insufficient for determining circulating hormone levels in the fetus. Hence, at ED16.5, hormonal content of the full-body (excluding gonads) was used to assess the impact of phthalate exposure. Hormone levels were quantified by ELISA. At ED16.5, full-body T level was lower in exposed females (2.93 vs. 12.23 pg/body in control, P<0.0001), but higher in exposed males (277 vs. 216 pg/body in control, P=0.022). However, there was no difference in fetal testicular T (871 vs. 1027 pg/2 testes in control, P=0.089), indicating that testicular steroidogenesis was unaltered. A rapid decline in serum T levels occurred in both newborn control (h1= 623 pg/mL, h6= 249 pg/mL, P=0.009) and exposed males (h1= 600 pg/mL, h6= 224 pg/mL, P=0.0015), but serum LH remained constant (Control: h1= 0.83 ng/mL, h6= 0.63 ng/mL, P=0.36; exposed: h1= 0.69 ng/mL, h6= 0.82 ng/mL, P=0.43). Phthalate exposure did not impact neonatal serum T or LH in males. However, at h6, exposed females had lower serum E2 than controls (0.7 vs. 8.51 pg/mL in control, P=0.001). This study found that prenatal phthalate exposure altered T and E2 levels of males and females distinctly, without disrupting LH secretion and gonadal steroidogenesis. The results indicate that T and E2 levels were likely disrupted by altered steroid clearance due to a sex-dependent impact on hepatic T and E2 metabolism, rather than pituitary or gonadal hormone production. In conclusion, prenatal exposure to a phthalate mixture alters the levels of T and E2 during fetal and neonatal EP in a gender-specific manner. Presentation: Thursday, June 15, 2023

Cryoablation of the adenohypophysis in the treatment of chronic pain syndrome in patients with stage IV malignant neoplasms

Background. The data of the European Society For Medical Oncology (ESMO) indicate that 64.0% of cancer patients with the fourth stage of cancer have chronic pain syndrome, which is the main factor that significantly affects the quality of life. In 46.0% of patients, it is not possible to obtain a stable analgesic effect with modern methods of analgesia, including interventional methods and pharmacotherapy. Refractory oncological pain stimulates the search for new methods of pain relief.
 Purpose – retrospective assessment of the effectiveness of the selective stereo- tactic transnasal transsphenoidal cryoablation of the adenohypophysis in the treatment of chronic pain syndrome in patients with stage IV malignant neoplasms.
 Materials and methods. 45 microsurgeries were performed – stereotactic selective transnasal transsphenoidal cryoablation of the adenohypophysis with endoscopy. Over the period from 2014 to 2018, 45 patients with stage IV malignant hormone- dependent neoplasms of various somatic organs and chronic pain syndrome underwent microsurgery.
 Results. The analgesic effect appeared 4–6 hours after the microsurgery. In 95.6% of patients, an analgesic effect was achieved to a greater extent (according to the numeric rating scale (NRS) for pain, its intensity decreased from 7–9 points to 1–3 points); in other patients, the analgesic effect was achieved to a lesser extent (according to the NRS, pain intensity decreased from 7–9 points to 3–5 points), regardless of the condition and age of the patient. Accordingly, the dosage was reduced or opioid analgesics were discontinued. Severe complications after the cryoablation of adenohypophysis, such as meningitis, diabetes insipidus, and hypopituitary syndrome were not observed. In the postoperative period, three patients were treated for mild hypopituitary syndrome, and two patients were treated for liquorrhea with conservative therapy for 3–5 days.
 Conclusions. Stereotactic selective transnasal transsphenoidal cryoablation of the adenohypophysis is an effective microsurgery in the treatment of chronic pain syndrome in stage IV cancer patients. It improves the patient’s condition and quality of life. The analgesic effect appears in the first hours after surgery. Due to the use of cryoprobes with a diameter of 1.2 mm and 1.8 mm, the surgery is minimally traumatic and can be performed on patients in critical condition.

Pituitary crosstalk with bone, adipose tissue and brain.

Traditional textbook physiology has ascribed unitary functions to hormones from the anterior and posterior pituitary gland, mainly in the regulation of effector hormone secretion from endocrine organs. However, the evolutionary biology of pituitary hormones and their receptors provides evidence for a broad range of functions in vertebrate physiology. Over the past decade, we and others have discovered that thyroid-stimulating hormone, follicle-stimulating hormone, adrenocorticotropic hormone, prolactin, oxytocin and arginine vasopressin act directly on somatic organs, including bone, adipose tissue and liver. New evidence also indicates that pituitary hormone receptors are expressed in brain regions, nuclei and subnuclei. These studies have prompted us to attribute the pathophysiology of certain human diseases, including osteoporosis, obesity and neurodegeneration, at least in part, to changes in pituitary hormone levels. This new information has identified actionable therapeutic targets for drug discovery.

Y chromosome toxicity does not contribute to sex-specific differences in longevity.

While sex chromosomes carry sex-determining genes, they also often differ from autosomes in size and composition, consisting mainly of silenced heterochromatic repetitive DNA. Even though Y chromosomes show structural heteromorphism, the functional significance of such differences remains elusive. Correlative studies suggest that the amount of Y chromosome heterochromatin might be responsible for several male-specific traits, including sex-specific differences in longevity observed across a wide spectrum of species, including humans. However, experimental models to test this hypothesis have been lacking. Here we use the Drosophila melanogaster Y chromosome to investigate the relevance of sex chromosome heterochromatin in somatic organs in vivo. Using CRISPR-Cas9, we generated a library of Y chromosomes with variable levels of heterochromatin. We show that these different Y chromosomes can disrupt gene silencing in trans, on other chromosomes, by sequestering core components of the heterochromatin machinery. This effect is positively correlated to the level of Y heterochromatin. However, we also find that the ability of the Y chromosome to affect genome-wide heterochromatin does not generate physiological sex differences, including sexual dimorphism in longevity. Instead, we discovered that it is the phenotypic sex, female or male, that controls sex-specific differences in lifespan, rather than the presence of a Y chromosome. Altogether, our findings dismiss the 'toxic Y' hypothesis that postulates that the Y chromosome leads to reduced lifespan in XY individuals.

Gene therapy for cross-correction of somatic organs and the CNS in mucopolysaccharidosis II in rodents and non-human primates

Mucopolysaccharidosis II (MPS II) is a rare lysosomal storage disease characterized by deficient activity of iduronate-2-sulfatase (I2S), leading to pathological accumulation of glycosaminoglycans (GAGs) in tissues. We used iduronate-2-sulfatase knockout (Ids KO) mice to investigate if liver-directed recombinant adeno-associated virus vectors (rAAV8-LSP-hIDSco) encoding human I2S (hI2S) could cross-correct I2S deficiency in Ids KO mouse tissues, and we then assessed the translation of mouse data to non-human primates (NHPs). Treated mice showed sustained hepatic hI2S production, accompanied by normalized GAG levels in somatic tissues (including critical tissues such as heart and lung), indicating systemic cross-correction from liver-secreted hI2S. Brain GAG levels in Ids KO mice were lowered but not normalized; higher doses were required to see improvements in brain histology and neurobehavioral testing. rAAV8-LSP-hIDSco administration in NHPs resulted in sustained hepatic hI2S production and therapeutic hI2S levels in cross-corrected somatic tissues but no hI2S exposure in the central nervous system, perhaps owing to lower levels of liver transduction in NHPs than in mice. Overall, we demonstrate the ability of rAAV8-LSP-hIDSco to cross-correct I2S deficiency in mouse somatic tissues and highlight the importance of showing translatability of gene therapy data from rodents to NHPs, which is critical for supporting translation to clinical development.

Self Assessments of the Quality of Own Life during a Viral Pandemic and War

This publication is the result of a study of the dynamics of self-assessment of quality of life indicators by young medical workers (students) against the background of the negative impact of the infodemic due to the chronic stress situation during the covid 19 pandemic, quarantine restrictions and professional communication with various patients, and the full-scale war of complete destruction that began on 24.02.22 all citizens of Ukraine. General clinical methods were used as research methods - to assess the condition of the mucous membranes, skin and somatic organs, as well as psychodiagnostic questionnaire SF36 for anonymous self-determination of the quality of one's life. The analysis of the obtained results of two hundred young medical students during 2021-2022 revealed a number of correlational dependencies from the amount of time during which they are forced to study and work against the background of the mostly negative impact of the infodemic, initially caused only by the situation of the viral pandemic and the need daily contact with various patients, and after 02.24.22 and in conditions real large-scale war and martial law. That is, already in conditions of real danger of being injured or killed during another missile attack in the rear and being infected with another viral strain of coronavirus infection due to forced daily professional contact with various somatic patients. The awareness of the presence of these two factors directly reduces their self-esteem of the quality of their own life and is, in certain situations, a sufficient factor that contributes to the formation of psychosomatic disorders as a manifestation of the initial clinical signs of somatic-emotional professional pathology, which, in particular, manifests itself in the appearance of various rashes on the skin

Sanfilippo Syndrome: Optimizing Care with a Multidisciplinary Approach.

Sanfilippo syndrome, or mucopolysaccharidosis type III (MPS III), is a disease grouping five genetic disorders, four of them occurring in humans and one known to date only in a mouse model. In every subtype of MPS III (designed A, B, C, D or E), a lack or drastically decreased activity of an enzyme involved in the degradation of heparan sulfate (HS) (a compound from the group of glycosaminoglycans (GAGs)) arises from a genetic defect. This leads to primary accumulation of HS, and secondary storage of other compounds, combined with changes in expressions of hundreds of genes and many defects in organelles and various biochemical processes in the cell. As a result, dysfunctions of tissues and organs occur, leading to severe symptoms in patients. Although changes in somatic organs are considerable, the central nervous system is especially severely affected, and neurological, cognitive and behavioral disorders are the most significant changes, making the disease enormously burdensome for patients and their families. In the light of the current lack of any registered therapy for Sanfilippo syndrome (despite various attempts of many research groups to develop effective treatment, still no specific drug or procedure is available for MPS III), optimizing care with a multidisciplinary approach is crucial for managing this disease and making quality of patients’ life passable. This includes efforts to make/organize (i) accurate diagnosis as early as possible (which is not easy due to various possible misdiagnosis events caused by similarity of MPS III symptoms to those of other diseases and variability of patients), (ii) optimized symptomatic treatment (which is challenging because of complexity of symptoms and often untypical responses of MPS III patients to various drugs), and (iii) psychological care (for both patients and family members and/or caregivers). In this review article, we focus on these approaches, summarizing and discussing them.

Introducing Large Genomic Deletions in Human Pluripotent Stem Cells Using CRISPR-Cas3.

CRISPR-Cas systems provide researchers with eukaryotic genome editing tools and therapeutic platforms that make it possible to target disease mutations in somatic organs. Most of these tools employ Type II (e.g., Cas9) or Type V (e.g., Cas12a) CRISPR enzymes to create RNA-guided precise double-strand breaks in the genome. However, such technologies are limited in their capacity to make targeted large deletions. Recently, the Type I CRISPR system, which is prevalent in microbes and displays unique enzymatic features, has been harnessed to effectively create large chromosomal deletions in human cells. Type I CRISPR first uses a multisubunit ribonucleoprotein (RNP) complex called Cascade to find its guide-complementary target site, and then recruits a helicase-nuclease enzyme, Cas3, to travel along and shred the target DNA over a long distance with high processivity. When introduced into human cells as purified RNPs, the CRISPR-Cas3 complex can efficiently induce large genomic deletions of varying lengths (1-100 kb) from the CRISPR-targeted site. Because of this unique editing outcome, CRISPR-Cas3 holds great promise for tasks such as the removal of integrated viral genomes and the interrogation of structural variants affecting gene function and human disease. Here, we provide detailed protocols for introducing large deletions using CRISPR-Cas3. We describe step-by-step procedures for purifying the Type I-E CRISPR proteins Cascade and Cas3 from Thermobifida fusca, electroporating RNPs into human cells, and characterizing DNA deletions using PCR and sequencing. We focus here on human pluripotent stem cells due to their clinical potential, but these protocols will be broadly useful for other cell lines and model organisms for applications including large genomic deletion, full-gene or -chromosome removal, and CRISPR screening for noncoding elements, among others. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Expression and purification of Tfu Cascade RNP Support Protocol 1: Expression and purification of TfuCas3 protein Support Protocol 2: Culture of human pluripotent stem cells Basic Protocol 2: Introduction of Tfu Cascade RNP and Cas3 protein into hPSCs via electroporation Basic Protocol 3: Characterization of genomic DNA lesions using long-range PCR, TOPO cloning, and Sanger sequencing Alternate Protocol: Comprehensive analysis of genomic lesions by Tn5-based next-generation sequencing Support Protocol 3: Single-cell clonal isolation.

Asymmetric Phenotypes of Sterile Hybrid Males From Reciprocal Crosses Between Species of the Anopheles gambiae Complex

Haldane’s rule of speciation states that sterility or inviability affects the heterogametic sex of inter-species hybrids. Darwin’s corollary to Haldane’s rule implies that there are asymmetric phenotypes in inter-species hybrids from reciprocal crosses. Studying the phenotypes of F1 hybrids among closely related species of malaria mosquitoes can assist researchers in identifying the genetic factors and molecular mechanisms of speciation. To characterize phenotypes of sterile hybrid males in the Anopheles gambiae complex, we performed crosses between laboratory strains of An. merus and either An. gambiae or An. coluzzii. The reproductive tracts had normal external morphology in hybrid males from crosses between female An. merus and male An. gambiae or An. coluzzii. Despite being sterile, these males could copulate with females for a normal period of time and could transfer a mating plug to induce female oviposition and monogamy. In contrast, the entire reproductive tracts in hybrid males from crosses between female An. gambiae or An. coluzzii and male An. merus were severely underdeveloped. These males had atrophic testes and reduced somatic organs of the reproductive system including male accessary glands and ejaculatory duct. In addition, hybrid males with underdeveloped reproductive tracts displayed a shorter copulation time with females and failed to induce female oviposition and monogamy due to their inability to form and transfer a plug to females during mating. The asymmetry of the phenotypes associated with hybrid male sterility suggests that different genetic factors and molecular mechanisms are responsible for reproductive isolation in reciprocal crosses among species of the An. gambiae complex.

Influence of standard and stimulated growth of B16/F10 melanoma on AIF levels in mitochondria in cells of the heart and other somatic organs in female mice

studying AIF levels in mitochondria in cells of the heart and various organs in female mice with the growth of experimental melanoma B16 / F10 and comorbid pathology

Long-lived termite kings and queens activate telomerase in somatic organs.

Kings and queens of termites, like queens of other advanced eusocial insects, are endowed with admirable longevity, which dramatically exceeds the life expectancies of their non-reproducing nest-mates and related solitary insects. In the quest to find the mechanisms underlying the longevity of termite reproductives, we focused on somatic maintenance mediated by telomerase. This ribonucleoprotein is well established for pro-longevity functions in vertebrates, thanks primarily to its ability of telomere extension. However, its participation in lifespan regulation of insects, including the eusocial taxa, remains understudied. Here, we report a conspicuous increase of telomerase abundance and catalytic activity in the somatic organs of primary and secondary reproductives of the termite Prorhinotermes simplex and confirm a similar pattern in two other termite species. These observations stand in contrast with the telomerase downregulation characteristic for most adult somatic tissues in vertebrates and also in solitary insects and non-reproducing castes of termites. At the same time, we did not observe caste-specific differences in telomere lengths that might explain the differential longevity of termite castes. We conclude that although the telomerase activation in termite reproductives is in line with the broadly assumed association between telomerase and longevity, its direct phenotypic impact remains to be elucidated.

Caffeine protects against stress-induced murine depression through activation of PPAR\u03b3C1\u03b1-mediated restoration of the kynurenine pathway in the skeletal muscle

Exercise prevents depression through peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α)-mediated activation of a particular branch of the kynurenine pathway. From kynurenine (KYN), two independent metabolic pathways produce neurofunctionally different metabolites, mainly in somatic organs: neurotoxic intermediate metabolites via main pathway and neuroprotective end product, kynurenic acid (KYNA) via the branch. Elevated levels of KYN have been found in patients with depression. Herein, we investigated whether and how caffeine prevents depression, focusing on the kynurenine pathway. Mice exposed to chronic mild stress (CMS) exhibited depressive-like behaviours with an increase and decrease in plasma levels of pro-neurotoxic KYN and neuroprotective KYNA, respectively. However, caffeine rescued CMS-exposed mice from depressive-like behaviours and restored the plasma levels of KYN and KYNA. Concomitantly, caffeine induced a key enzyme converting KYN into KYNA, namely kynurenine aminotransferase-1 (KAT1), in murine skeletal muscle. Upon caffeine stimulation murine myotubes exhibited KAT1 induction and its upstream PGC-1α sustainment. Furthermore, a proteasome inhibitor, but not translational inhibitor, impeded caffeine sustainment of PGC-1α, suggesting that caffeine induced KAT1 by inhibiting proteasomal degradation of PGC-1α. Thus, caffeine protection against CMS-induced depression may be associated with sustainment of PGC-1α levels and the resultant KAT1 induction in skeletal muscle, and thereby consumption of pro-neurotoxic KYN.

The male germline-specific protein MAPS is indispensable for pachynema progression and fertility

Meiosis is a specialized cell division that creates haploid germ cells from diploid progenitors. Through differential RNA expression analyses, we previously identified a number of mouse genes that were dramatically elevated in spermatocytes, relative to their very low expression in spermatogonia and somatic organs. Here, we investigated in detail 1700102P08Rik, one of these genes, and independently conclude that it encodes a male germline-specific protein, in agreement with a recent report. We demonstrated that it is essential for pachynema progression in spermatocytes and named it male pachynema-specific (MAPS) protein. Mice lacking Maps (Maps-/- ) suffered from pachytene arrest and spermatocyte death, leading to male infertility, whereas female fertility was not affected. Interestingly, pubertal Maps-/- spermatocytes were arrested at early pachytene stage, accompanied by defects in DNA double-strand break (DSB) repair, crossover formation, and XY body formation. In contrast, adult Maps-/- spermatocytes only exhibited partially defective crossover but nonetheless were delayed or failed in progression from early to mid- and late pachytene stage, resulting in cell death. Furthermore, we report a significant transcriptional dysregulation in autosomes and XY chromosomes in both pubertal and adult Maps-/- pachytene spermatocytes, including failed meiotic sex chromosome inactivation (MSCI). Further experiments revealed that MAPS overexpression in vitro dramatically decreased the ubiquitination levels of cellular proteins. Conversely, in Maps-/- pachytene cells, protein ubiquitination was dramatically increased, likely contributing to the large-scale disruption in gene expression in pachytene cells. Thus, MAPS is a protein essential for pachynema progression in male mice, possibly in mammals in general.

Spinal neuron-glia-immune interaction in cross-organ sensitization

Inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS), historically considered as regional gastrointestinal disorders with heightened colonic sensitivity, are increasingly recognized to have concurrent dysfunction of other visceral and somatic organs, such as urinary bladder hyperactivity, leg pain, and skin hypersensitivity. The interorgan sensory cross talk is, at large, termed "cross-organ sensitization." These organs, anatomically distant from one another, physiologically interlock through projecting their sensory information into dorsal root ganglia (DRG) and then the spinal cord for integrative processing. The fundamental question of how sensitization of colonic afferent neurons conveys nociceptive information to activate primary afferents that innervate distant organs remains ambiguous. In DRG, primary afferent neurons are surrounded by satellite glial cells (SGCs) and macrophage accumulation in response to signals of injury to form a neuron-glia-macrophage triad. Astrocytes and microglia are major resident nonneuronal cells in the spinal cord to interact, physically and chemically, with sensory synapses. Cumulative evidence gathered so far indicate the indispensable roles of paracrine/autocrine interactions among neurons, glial cells, and immune cells in sensory cross-activation. Dichotomizing afferents, sensory convergency in the spinal cord, spinal nerve comingling, and extensive sprouting of central axons of primary afferents each has significant roles in the process of cross-organ sensitization; however, more results are required to explain their functional contributions. DRG that are located outside the blood-brain barrier and reside upstream in the cascade of sensory flow from one organ to the other in cross-organ sensitization could be safer therapeutic targets to produce less central adverse effects.

HSPA2 Chaperone Contributes to the Maintenance of Epithelial Phenotype of Human Bronchial Epithelial Cells but Has Non-Essential Role in Supporting Malignant Features of Non-Small Cell Lung Carcinoma, MCF7, and HeLa Cancer Cells.

Simple SummaryHeat shock proteins A (HSPA) are molecular chaperones that play a central role in the cellular protein quality control. One of them is HSPA2 which recently was indicated as a novel cancer-related protein due to its elevated expression in various tumors and reported prognostic significance. Several previous in vitro studies have shown significant role of HSPA2 in supporting cancer cells growth and invasiveness. Our data presented in this article contradict the current belief of the essential role of HSPA2 chaperone and show that HSPA2 is not crucial for maintenance of the malignant phenotype of lung, breast, and cervical cancer cells. Instead, we revealed HSPA2’s role in supporting clonogenic potential and adhesive ability of bronchial epithelial cells. Therefore, further research should concentrate on elucidating HSPA2 roles in epithelial cells.Heat Shock Protein A2 (HSPA2) is a member of the HSPA (HSP70) chaperone family and has a critical role for male fertility. HSPA2 is present in a number of somatic organs. Limited evidence suggests that HSPA2 may be involved in regulating epithelial cell differentiation. HSPA2 also emerged as a cancer-related chaperone; however, no consensus on its functional significance has been reached so far. In this study, we compared the phenotypic effects of HSPA2 deficit in non-transformed human bronchial epithelial cells (HBEC), and in lung, breast, and cervical cancer cells. We used various techniques to inhibit the HSPA2 gene expression in order to examine the impact of HSPA2 deficiency on cell growth, migration, adhesion, and invasion. Our results show that HBEC but not cancer cells are sensitive to HSPA2 deficit. HSPA2 knockdown in HBEC cells impaired their clone-forming ability and adhesiveness. Thus, our results indicate that epithelial cells can rely on a specific activity of HSPA2, but such dependence can be lost in epithelial cells that have undergone malignant transformation.

Cardiorenal metabolic biomarkers link early life stress to risk of non-communicable diseases and adverse mental health outcomes

Stress is one of the most critical determinants of lifetime health and increases the risk of chronic non-communicable diseases. To gain insight into underlying environment-gene interactions, we analyzed the cardiorenal metabolome of adult mice exposed to multidimensional early-life transportation stress. Using proton nuclear magnetic resonance (1H NMR) spectroscopy, we show that early life stress permanently programs metabolic pathways in somatic organs linked to cardiorenal and mental health disorders in later life. Heart and kidneys of stressed mice revealed robust metabolic markers linked to abnormal energy metabolism, branched-chain amino acid biosynthesis and degradation, methylhistidine metabolism, arginine and proline metabolism, glycine and serine metabolism, and aminoacyl-tRNA biosynthesis. These markers were strongly associated with anxiety-like behaviours. Dysregulation of energy and protein metabolism suggests an increased risk of metabolic diseases like insulin resistance, cardiorenal syndrome, diabetes, and obesity. These findings provide novel insights into the direct effects of early life stress on cardiorenal metabolism and are consistent with prior observations of increased non-communicable disease risk in stressed populations. Thus, stress-associated metabolic signatures in somatic organs may provide early predictors of health risks in later life and reveal new candidates for peripheral biomarker detection with diagnostic value.

Investigation of the Effect of Pulsatile and Nonpulsatile Flow on Kidney in Coronary Surgery With NIRS.

Acute renal insufficiency is a significant cause of morbidity and mortality after coronary artery bypass grafting performed with cardiopulmonary bypass. Functional near-infrared spectroscopy (fNIRS) is an emerging brain-imaging technique that can be used to detect organ perfusions in adults. This study aims to determine the effects of pulsatile and nonpulsatile flow on renal circulation by using functional near-infrared spectroscopy and biochemical markers. Forty patients, who had undergone isolated CABG between March 2014 and July 2014 in the authors' clinic, were included in the study. Patients were divided in the pulsatile and nonpulsatile groups by simple randomization. Urine outputs statistically were higher in the pulsatile group, during CPB (P = .045). Renal perfusion measurements via fNIRS intra-operatively statistically were parallel between the groups. When we compared biochemical markers within the groups, in the nonpulsatile group, creatinin statistically is higher on the postoperative first day (P = .003), and BUN statistically is higher on the postoperative first (P = .015), second (P = .001), and fifth (P = .020) days, according to preoperative value. In the pulsatile group, only the postoperative second day BUN is higher (P = .007). fNIRS is being used to track cerebral functions. During operation, it also provides a correct observation for blood feeding of somatic organs, such as the kidneys, but it requires more clinical study to be accepted as routine.

Clinical and genetic features of the formation of concomitant visceral pathology in workers with industrial fluorosis

Introduction. The urgency of the problem determines the high level of occupational morbidity in aluminum production workers and the prevalence of somatic pathology, which occupies one of the leading positions in the pathogenesis of fluorosis.The aim of the study is to determine the clinical and genetic features of the nature of concomitant visceral pathology in workers with industrial fluorosis.Materials and methods. A complex of clinical, biochemical, molecular, genetic, and instrumental research methods was used in patients with fluorosis (246 people) and in the control group (106 people).Results. A high frequency of health disorders in workers engaged in the production of aluminum was determined during long-term monitoring. An extended clinical and genetic study of the state of somatic organs in workers with fluorosis and in control is presented. It is revealed that in the group of subjects with proven diagnosis, pathologies concomitant with the main disease were significantly more common: pathologies of musculoskeletal and articular systems, blood circulation, respiratory (rhinitis, chronic, toxic and dust-borne bronchitis, chronic obstructive pulmonary disease), hepatobiliary (steatosis, fluoride hepatopathy), gastrointestinal tract (chronic gastritis, cholecystopancreatitis), renal systems (pyelonephritis, urolithiasis), as well as a combination of several nosologies was in 43%. The association of GG VEGF genotypes with chronic pyelonephritis and TT Il1ß, GSTT 1 0/0 with fluoride hepatosis was revealed in patients with fluorosis.Conclusions. The structure of occupational morbidity in workers of the main professions of aluminum production is dominated by concomitant pathology of functional systems against the background of bone destruction, determined by endogenous factors of the course of metabolic processes of the body.The authors declare no conflict of interests.

Investigating the role of BCAR4 in ovarian physiology and female fertility by genome editing in rabbit

Breast Cancer Anti-estrogen Resistance 4 (BCAR4) was previously characterised in bovine species as a gene preferentially expressed in oocytes, whose inhibition is detrimental to in vitro embryo development. But its role in oogenesis, folliculogenesis and globally fertility in vivo remains unknown. Because the gene is not conserved in mice, rabbits were chosen for investigation of BCAR4 expression and function in vivo. BCAR4 displayed preferential expression in the ovary compared to somatic organs, and within the ovarian follicle in the oocyte compared to somatic cells. The transcript was detected in follicles as early as the preantral stage. Abundance decreased throughout embryo development until the blastocyst stage. A lineage of genome-edited rabbits was produced; BCAR4 expression was abolished in follicles from homozygous animals. Females of wild-type, heterozygous and homozygous genotypes were examined for ovarian physiology and reproductive parameters. Follicle growth and the number of ovulations in response to hormonal stimulation were not significantly different between genotypes. Following insemination, homozygous females displayed a significantly lower delivery rate than their heterozygous counterparts (22 ± 7% vs 71 ± 11% (mean ± SEM)), while prolificacy was 1.8 ± 0.7 vs 6.0 ± 1.4 kittens per insemination. In conclusion, BCAR4 is not essential for follicular growth and ovulation but it contributes to optimal fertility in rabbits.