Deficient Background Research Articles

Abstract 2516: Loss of cadherin 11 in pancreatic cancer induces altered immune cell infiltration

Abstract Pancreatic ductal adenocarcinoma (PDAC) is one of the top five deadliest forms of cancer with very few treatment options. The 5-year survival rate for PDAC is 10% following diagnosis. Preclinical murine models have been developed that leverage key driver mutations and have significantly contributed to our understanding of PDAC. One such genetically engineered mouse model (GEMM) that has emerged as an important tool is the KPC mouse (LSL-KrasG12D/+;LSL-Trp53R172H/+; p48-Cre) that spontaneously develops pancreatic tumors at ~14-16 weeks of age. Cadherin-11 (Cdh11), a cell-to-cell adhesion molecule has been suggested to play a role in development of the desmoplastic stroma in PDAC, that leads to difficulties in drug accessibility and has been hypothesized to contribute to chemotherapeutic resistance and correlate with poor prognosis. However, the mechanisms by which Cdh11 deficiency in the stromal microenvironment of PDAC-bearing KPC mice influences tumor infiltrating immune cells, has yet to be fully understood. Single-cell RNA sequencing (scRNAseq) of the immune (CD45+) compartment of tumor bearing Cdh11 proficient (KPC/Cdh11+/+), tumor bearing Cdh11 deficient (KPC/Cdh11+/-), non-tumor bearing Cdh11 deficient (Cdh11+/-) and wildtype (Cdh11+/+) mice was performed. We observed a sharp decrease in the presence of myeloid/monocyte lineage cells (CD14+) in KPC/Cdh11+/- tumors and also an increase in T, B and plasma cells, compared to KPC/Cdh11+/+ tumors. Genes upregulated in infiltrating T- and NK cells specific to a Cdh11 deficient background include Cd8a, Nkg7, Maf. Additionally, genes found to be upregulated in B cell clusters in Cdh11 deficient mice include those related to B cell differentiation/activation such as Lgals1, Id2, Itgb1, Rgs1. The increase in B and T cell infiltration was specific to the Cdh11 deficient background, since both pancreata from KPC/Cdh11+/+and Cdh11+/- mice had elevated levels of infiltration. Immunohistochemical validation of these findings has confirmed these changes in tumor infiltrating immune cells. Additionally of note, an increase in antibody-producing plasma cells was observed specifically in a Cdh11 deficient background. Igkc, an immunoglobulin found to be enriched in plasma cells was highly expressed in this group of immune cells. We also observed that KPC/Cdh11+/-had significantly more Igkc expressing cells than KPC/Cdh11+/+. Future work is needed to clearly define the role of Cdh11 in modulating B, T and plasma cell behavior and subsequent contributions to PDAC outcome. This study received funding by LDRD 19-SI-003. This work was conducted under the auspices of the USDOE by LLNL (DE-AC52-07NA27344), LLNL-ABS-820889. This work was supported by AACR-AstraZeneca Fellowship in Immunooncology Research, grant 17-40-12-PERA; The Ruesch Center for the Cure of Gastrointestinal Cancers grant award; NIH R01 CA170653; and NIH Cancer Center Support Grant P30 CA051008. Citation Format: Kelly A. Martin, Aimy Sebastian, Nicholas Hum, Ivana Peran, Stephen Byers, Elizabeth Wheeler, Matthew Coleman, Gabriela Loots. Loss of cadherin 11 in pancreatic cancer induces altered immune cell infiltration [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2516.

Myeloid Cell PKM2 Deletion Enhances Efferocytosis and Reduces Atherosclerosis.

The glycolytic enzyme PKM2 (pyruvate kinase muscle 2) is upregulated in monocytes/macrophages of patients with atherosclerotic coronary artery disease. However, the role of cell type-specific PKM2 in the setting of atherosclerosis remains to be defined. We determined whether myeloid cell-specific PKM2 regulates efferocytosis and atherosclerosis. We generated myeloid cell-specific PKM2-/- mice on Ldlr (low-density lipoprotein receptor)-deficient background (PKM2mye-KOLdlr-/-). Controls were littermate PKM2WTLdlr-/- mice. Susceptibility to atherosclerosis was evaluated in whole aortae and cross sections of the aortic sinus in male and female mice fed a high-fat Western diet for 14 weeks, starting at 8 weeks. PKM2 was upregulated in macrophages of Ldlr-/- mice fed a high-fat Western diet compared with chow diet. Myeloid cell-specific deletion of PKM2 led to a significant reduction in lesions in the whole aorta and aortic sinus despite high cholesterol and triglyceride levels. Furthermore, we found decreased macrophage content in the lesions of myeloid cell-specific PKM2-/- mice associated with decreased MCP-1 (monocyte chemoattractant protein 1) levels in plasma, reduced transmigration of macrophages in response to MCP-1, and impaired glycolytic rate. Macrophages isolated from myeloid-specific PKM2-/- mice fed the Western diet exhibited reduced expression of proinflammatory genes, including MCP-1, IL (interleukin)-1β, and IL-12. Myeloid cell-specific PKM2-/- mice exhibited reduced apoptosis concomitant with enhanced macrophage efferocytosis and upregulation of LRP (LDLR-related protein)-1 in macrophages in vitro and atherosclerotic lesions in vivo. Silencing LRP-1 in PKM2-deficient macrophages restored inflammatory gene expression and reduced efferocytosis. As a therapeutic intervention, inhibiting PKM2 nuclear translocation using a small molecule reduced glycolytic rate, enhanced efferocytosis, and reduced atherosclerosis in Ldlr-/- mice. Genetic deletion of PKM2 in myeloid cells or limiting its nuclear translocation reduces atherosclerosis by suppressing inflammation and enhancing efferocytosis.

Targeting ApoC3 Paradoxically Aggravates Atherosclerosis in Hamsters With Severe Refractory Hypercholesterolemia.

RationaleApoC3 plays a central role in the hydrolysis process of triglyceride (TG)-rich lipoproteins mediated by lipoprotein lipase (LPL), which levels are positively associated with the incidence of cardiovascular disease (CVD). Although targeting ApoC3 by antisense oligonucleotide (ASO), Volanesorsen markedly reduces plasma TG level and increase high-density lipoprotein cholesterol (HDL-C) in patients with hypertriglyceridemia (HTG), the cholesterol-lowering effect of ApoC3 inhibition and then the consequential outcome of atherosclerotic cardiovascular disease (ASCVD) have not been reported in patients of familial hypercholesterolemia (FH) with severe refractory hypercholesterolemia yet.ObjectiveTo investigate the precise effects of depleting ApoC3 on refractory hypercholesterolemia and atherosclerosis, we crossed ApoC3-deficient hamsters with a background of LDLR deficiency to generate a double knockout (DKO) hamster model (LDLR−/−, XApoC3−/−, DKO).Approach and ResultsOn the standard laboratory diet, DKO hamsters had reduced levels of plasma TG and total cholesterol (TC) relative to LDLR−/− hamsters. However, upon high-cholesterol/high-fat (HCHF) diet feeding for 12 weeks, ApoC3 deficiency reduced TG level only in female animals without affecting refractory cholesterol in the circulation, whereas apolipoprotein A1 (ApoA1) levels were significantly increased in DKO hamsters with both genders. Unexpectedly, loss of ApoC3 paradoxically accelerated diet-induced atherosclerotic development in female and male LDLR−/− hamsters but ameliorated fatty liver in female animals. Further analysis of blood biological parameters revealed that lacking ApoC3 resulted in abnormal platelet (PLT) indices, which could potentially contribute to atherosclerosis in LDLR−/− hamsters.ConclusionsIn this study, our novel findings provide new insight into the application of ApoC3 inhibition for severe refractory hypercholesterolemia and ASCVD.

Effect of Acetaminophen Against the Background of Alimentary Protein Deficiency on the Features of Sulfur-Containing Amino Acids Metabolism in Rats

Effect of Acetaminophen Against the Background of Alimentary Protein Deficiency on the Features of Sulfur-Containing Amino Acids Metabolism in Rats

PYRUVATE DEHYDROGENASE COMPLEX ACTIVITY IN THE RAT`S KIDNEYS UNDER CONDITIONS OF TOXIC ACETAMINOPHEN DAMAGE WITH PROTEIN DEFICIENCY

The aim of this work was to study the research the enzymatic activity of pyruvate dehydrogenase complex and lac-tate dehydrogenase and determination of the lactate/pyruvate ratio in rat kidneys under conditions of toxic damage by acetaminophen against the background of protein deficiency.The activity of the pyruvate dehydrogenase complex activity was determined according to a method based on the reaction of oxidative decarboxylation of pyruvate with simultaneous reduction of NAD+, which is measured spectrophotometrically at 340 nm. LDH activity was studied by an optimized optical method, which is based on the reaction of conversion of pyruvate to lactate with concomitant oxidation of NADH, which was measured using a spectrophotometer at λ = 340 nm. Lactate content was measured spectrophotometrically by the ability of FeCl3 to interact with lactate ions, resulting in the formation of a bright yellow iron lactate solution, which is recorded at a wavelength of 390 nm. The concentration of pyruvate was deter-mined by the modified Umbright method, which is based on the reaction of the interaction of pyruvate with 2,4-Dinitrophenylhydrazine (2,4-DNPH) in an alkaline medium, during which hydrazine is formed, which was recorded using a photoelectrocolorimeter at λ = 440 nm. The study was conducted on 4 groups of experimental animals: I - control animals (K); II – rats kept on a low-protein diet (LPD); ІІІ – animals in which acute toxic damage was caused by acetaminophen (TI); IV – rats on a low-protein diet, simulated acute toxic damage with acetaminophen (LPD/TI). It is demonstrated that under the conditions of dietary protein deficiency in the kidneys, there is a decrease in the activi-ty of the pyruvate dehydrogenase complex while maintaining the control indicators of the lactate/pyruvate ratio and lactate dehydrogenase activity. However, under the conditions of the toxic effect of acetaminophen against the back-ground of dietary protein deficiency in the kidneys, an intensification of the anaerobic pathway of energy supply is observed, as evidenced by a significant decrease in the activity of the pyruvate dehydrogenase complex, an increase in the lactate/pyruvate ratio against the background of lactate dehydrogenase activation. The obtained results can be used to justify approaches to correct energy deficit in animals under the conditions of acetaminophen overdose against the background of protein deficiency.

PECULIARITIES OF CELLULAR IMMUNE REACTIONS IN WOMEN WITH CHRONIC GASTROINTESTINAL DISEASES IN THE STAGE OF LONG REMISSION IN THE ARCTIC LIVING AREA

Ecological and physiological studies conducted in recent years show that one of the most important factors determining the state of health is the environment. Natural and climatic factors of the Arctic zone contribute to a decrease in the reserve capacity of human immune homeostasis, the reduction of which forms secondary environmentally dependent immune imbalances, manifested by chronic pathology and frequent recurrences of diseases in the anamnesis, in particular, erosive gastritis. The study of the adaptive immune response during the period of long-term remission is extremely important, it allows us to predict the recurrence of the disease and carry out timely prevention. The study involved 30 women, aged 50–70 years with a history of chronic gastritis in the stage of long-term remission (3–5 years), residents of the Arctic region of the village. Pinega Arkhangelsk region. The study of the immune status included the determination in the peripheral blood of: the content of lymphocytes with CD3⁺, CD5⁺, CD8⁺, CD16⁺, CD25⁺, CD95⁺ receptors; phagocytic activity, phagocytic number using the application package Statistica 10.0, Microsoft Excel 2010. It was found that elevated concentrations of natural killer cells (CD16⁺) and cytotoxic cells (CD8⁺) occurred in 28,57±1,77% and 60,00±2,56% of cases, respectively, against the background of a pronounced deficiency in the content of T-lymphocytes (CD5⁺) in 96,66±3,26% and (CD3⁺) in 96,29±3,25% of the subjects, which indicates a reduction in the reserve capacity of immune homeostasis. Deficiency of phagocytic activity was registered in 23,33±1,60% of individuals. The revealed increased concentrations of cells with CD25⁺ and CD95⁺ receptors, combined with a high activity of phagocytosis, in our opinion, should be considered a compensatory mechanism of the adaptive immune response with a favorable prognosis for maintaining a long-term remission of the disease.

GDF15 Suppresses Lymphoproliferation and Humoral Autoimmunity in a Murine Model of Systemic Lupus Erythematosus

Growth and differentiation factor 15 (GDF15), a divergent member of the transforming growth factor-β superfamily, has been associated with acute and chronic inflammatory conditions including autoimmune disease, i.e., type I diabetes and rheumatoid arthritis. Still, its role in systemic autoimmune disease remains elusive. Thus, we studied GDF15-deficient animals in Fas-receptor intact (C57BL/6) or deficient (C57BL/6lpr/lpr) backgrounds. Further, lupus nephritis (LN) microdissected kidney biopsy specimens were analyzed to assess the involvement of GDF15 in human disease. GDF15-deficiency in lupus-prone mice promoted lymphoproliferation, T-, B- and plasma cell-expansion, a type I interferon signature, and increased serum levels of anti-DNA autoantibodies. Accelerated systemic inflammation was found in association with a relatively mild renal phenotype. Splenocytes of phenotypically overall-normal Gdf15−/− C57BL/6 and lupus-prone C57BL/6lpr/lpr mice displayed increased in vitro lymphoproliferative responses or interferon-dependent transcription factor induction in response to the toll-like-receptor (TLR)-9 ligand CpG, or the TLR-7 ligand Imiquimod, respectively. In human LN, GDF15 expression was downregulated whereas type I interferon expression was upregulated in glomerular- and tubular-compartments versus living donor controls. These findings demonstrate that GDF15 regulates lupus-like autoimmunity by suppressing lymphocyte-proliferation and -activation. Further, the data indicate a negative regulatory role for GDF15 on TLR-7 and -9 driven type I interferon signaling in effector cells of the innate immune system.

Loss of function of GATA3 induces basal-like mammary tumors.

Purpose: GATA3 is a transcription factor essential for mammary luminal epithelial cell differentiation. Expression of GATA3 is absent or significantly reduced in basal-like breast cancers. Gata3 loss-of-function impairs cell proliferation, making it difficult to investigate the role of GATA3 deficiency in vivo. We previously demonstrated that CDK inhibitor p18INK4c (p18) is a downstream target of GATA3 and restrains mammary epithelial cell proliferation and tumorigenesis. Whether and how loss-of-function of GATA3 results in basal-like breast cancers remains elusive.Methods: We generated mutant mouse strains with heterozygous germline deletion of Gata3 in p18 deficient backgrounds and developed a Gata3 depleted mammary tumor model system to determine the role of Gata3 loss in controlling cell proliferation and aberrant differentiation in mammary tumor development and progression.Results: Haploid loss of Gata3 reduced mammary epithelial cell proliferation with induction of p18, impaired luminal differentiation, and promoted basal differentiation in mammary glands. p18 deficiency induced luminal type mammary tumors and rescued the proliferative defect caused by haploid loss of Gata3. Haploid loss of Gata3 accelerated p18 deficient mammary tumor development and changed the properties of these tumors, resulting in their malignant and luminal-to-basal transformation. Expression of Gata3 negatively correlated with basal differentiation markers in MMTV-PyMT mammary tumor cells. Depletion of Gata3 in luminal tumor cells also reduced cell proliferation with induction of p18 and promoted basal differentiation. We confirmed that expression of GATA3 and basal markers are inversely correlated in human basal-like breast cancers.Conclusions: This study provides the first genetic evidence demonstrating that loss-of-function of GATA3 directly induces basal-like breast cancer. Our finding suggests that basal-like breast cancer may also originate from luminal type cancer.

High-risk pregnancy: effectiveness of personalized pregravid preparation and positive perinatal experience

Research objective: to study the possibilities of improving perinatal results by optimizing pergravid preparation in women with high perinatal risk.Materials and methods. The first stage – a cross-sectional study – consisted in determining the vitamin D (VD) status and assessing the course of pregnancy in 459 women in the II and III trimester; the second stage was a randomized controlled study of women with VD deficiency in group IIA (planning stage, 54 women) and IIB (I trimester, 60 women), who received the vitamin and mineral complex (VMC) Pregna-5 with a high calcitriol content and perinatal results were evaluated. VD in the blood was determined by the enzyme-linked immunosorbent assay.Results. VD level was optimal only in 30.7% of pregnant women (group ІА), in 69.3% of women its insufficiency or deficiency was found (group ІB). The frequency of threatened termination (9.9 vs. 45.6%; p < 0.01), preeclampsia (2.0 vs. 11%; p < 0.05), intrauterine growth retardation (7% only in group IIB, p < 0.01) was significantly higher against the background of BD deficiency. Normalization of the VD level (groups IIA and IIB) after VMC administration was observed within 1–3 months. Pregnancy complications value was higher in the group where the VMC was started in the first trimester: early gestosis (15 vs. 28.57%; p < 0.05), the threat of termination (22.2 vs. 36.7%; p < 0.05), bacterial vaginosis (7.4 vs. 31.7%; p < 0.05), placental dysfunction (18.5 vs. 40.0%; p <0.01), placental pathology in group IIB was detected 2–3 times more often. The differences between the preeclampsia (1.9 vs. 6.7%; p > 0.05) and anemia (16.6 vs. 28.3%; p > 0.05) were insignificant. Analysis of the childbirth results showed a significantly higher frequency of cesarean sections in group IIB (38.3 vs. 22.2%; p < 0.05), the average weight of newborns was less (3299.11 ± 128 g vs. 3643.24 ± 136 g; p < 0.01). The most effective was the VMC start at the stage of pregnancy planning (F = 13.35482; p = 0.000016); a significant difference was revealed in the course of pregnancy in the groups where VMC started in the first, second and third trimesters of pregnancy (Q = 4.67, p = 0.00458).Conclusions. Timely donation of VMC with a high content of cholecalciferol in women with high perinatal risk may lead to a positive course of pregnancy. Further research is required to obtain a convincing evidence base.

EPIDEMIOLOGICAL ASSESSMENT OF DYMAMICS OF THE PREVALENCE AND INCIDENCE OF THE THYROID GLAND DISEASES IN UKRAINE AND CHERNIVTSI REGION

Objective – to assess the dynamics of the prevalence and incidence of the thyroid diseaseamong the adult population of Ukraine and Chernivtsi region during 2015-2020.Material and methods. A comparative analysis of official accounting and reportingstatistics of the Ministry of Health of Ukraine and the main indicators of the endocrinologyservice of Chernivtsi region on the prevalence and incidence of adult pathology of thethyroid gland over the past 5 years.Results. In Ukraine, the number of pathologies of the thyroid gland has increased,accounting for 48% of the total incidence of endocrine diseases. The most commonpathology is diffuse and nodular goitres. Their level in the western region, includingChernivtsi region, is higher than the national average. In Ukraine the incidence ofhypothyroidism has increased 20.3% for 5 years. Thyrotoxicosis increased 9.1%, andthe prevalence of thyroiditis increased 16.8%. The increase in the Chernivtsi region wasslightly lower. The high frequency of thyroid pathologies depends on the imbalance oftrace elements and vitamins in the diet against a background of iodine deficiency in thediet, environmental degradation and insufficient preventive measures.Conclusions. The analysis shows that in the western regions of Ukraine it is necessaryto apply a differentiated approach to the diagnosis, treatment and prevention of patientswith various thyroid diseases.

Immune Dysregulation in Patients With Chromosome 18q Deletions-Searching for Putative Loci for Autoimmunity and Immunodeficiency.

IntroductionAutoimmune disorders, IgA deficiency, and allergies seem to be common among individuals with 18q deletion syndrome [OMIM 601808]. We aimed to determine the prevalence, mechanism, and genetic background of autoimmunity, immune deficiency, and allergy in a cohort of patients with 18q deletions.Material and MethodsMedical registries and social media were used to recruit the patients. Microarray oligonucleotide comparative genomic hybridization (aCGH) (Agilent, Santa Clara, CA, USA) was performed in all patients to identify size and location of chromosome 18 deletion. Clinical evaluation and medical record collection were performed in each of the study participants. The history of autoimmune disorders, severe and/or recurrent infections, and symptoms of allergy were noted. Total immunoglobulin IgG, IgA, IgM, IgE, and IgG1-4 serum levels were measured using nephelometry and ELISA methods. Lymphocyte T subset phenotyping was performed in 24 subjects from 18q del cohort. To predict the most promising candidate genes, we used the ENDEAVOUR—a free web resource for gene prioritization.Results18q deletion was confirmed by means of array CGH analysis in 27 individuals, 15 (55.6%) females and 12 males, referred to the project by specialists in medical genetics, diabetology, or pediatric endocrinology between May 2015 and December 2019. The mean age at examination was 11.8 years (min–max: 4.0–33.5). Autoimmune disorders were present in 14/27 (51.8%) of the cohort. In eight of patients, symptoms of immune deficiency coexisted with autoimmunity. Allergy was reported in nine of 27 (33.4%) patients. Over 89% of patients presented with at list one type of immunoglobulin (IgA, IgM, IgG, IgE, and IgG1-4) deficiency and eight of 25 (32%) had abnormalities in at least two major immunoglobulin (IgG, IgA, IgM) measurements (CVID-like phenotype). Patients with 18q del exhibited a significantly decreased CD4, Treg FOXP3+, TregFOXP3+Helios+, and TemCD4 cell numbers in comparison with the control groups of 24 T1DM patients and 28 healthy controls.ConclusionsPatients with 18q deletions frequently suffer from autoimmune disorders, recurrent infections, and allergy due to immune dysregulation presenting with variable antibody deficiencies and T-regulatory cell deficiency (CD4+CD25+CD127lowFOXP3+). The spectrum of speculations regarding which gene might be responsible for such phenotype ranges from single gene haploinsufficiency to deletion of a cluster of immunogenes located distally to 18q21.

Abstract 9035: Scavenger Receptor-BI: Implications in Lipoprotein(a) Metabolism

Introduction: Lipoprotein(a) [Lp(a)] is a unique lipoprotein that has emerged as an independent risk factor for developing vascular disease and stroke. Hypothesis: We hypothesize that SR-BI is critical to the regulation of Lp(a) metabolism and that mutations in SR-BI contribute to the pathogenesis of CVD and stroke in relationship to Lp(a). Methods: Transgenic mice carrying the human Lp(a) [hLp(a)] on the SR-BI (mSR-BI) deficient mouse background, Lp(a) transgenic mice on the human SR-BI (hSR-BI) with and without the mSR-BI, and Lp(a) transgenic mice on the mutant hSR-BI with and without mSR-BI were used to determine the impact on Lp(a) catabolism and atherosclerosis. Results: We determined the levels of Lp(a) in wildtype and SR-BI KO mice, lipid levels and profiles and oxidized state of the Lp(a). In addition, studies on the impact of SR-BI and Lp(a) on atherosclerosis were examined. We showed that Lp(a) was elevated in SR-BI deficient mice and Lp(a) was reduced in hSR-BI tg mice, compared to wildtype SR-BI mice. Lp(a) mice with the mutant hSR-BI protein, showed significantly elevated Lp(a) levels. When Lp(a) levels were increased, Lp(a) was highly oxidized, lipid profiles were altered and mice develop significant atherosclerosis. We also showed that Lp(a) mice showed a higher incidence of hemorrhagic strokes and had worse outcomes following ischemic strokes. Conclusions: These studies will lay the ground work for both basic and translational studies to understand the impact of Lp(a) in CVD and stroke.

UBB+1 reduces amyloid-β cytotoxicity by activation of autophagy in yeast.

UBB+1 is a mutated version of ubiquitin B peptide caused by a transcriptional frameshift due to the RNA polymerase II “slippage”. The accumulation of UBB+1 has been linked to ubiquitin-proteasome system (UPS) dysfunction and neurodegeneration. Alzheimer’s disease (AD) is defined as a progressive neurodegeneration and aggregation of amyloid-β peptides (Aβ) is a prominent neuropathological feature of AD. In our previous study, we found that yeast cells expressing UBB+1 at lower level display an increased resistance to cellular stresses under conditions of chronological aging. In order to examine the molecular mechanisms behind, here we performed genome-wide transcriptional analyses and molecular/cellular biology assays. We found that low UBB+1 expression activated the autophagy pathway, increased vacuolar activity, and promoted transport of autophagic marker ATG8p into vacuole. Furthermore, we introduced low UBB+1 expression to our humanized yeast AD models, that constitutively express Aβ42 and Aβ40 peptide, respectively. The co-expression of UBB+1 with Aβ42 or Aβ40 peptide led to reduced intracellular Aβ levels, ameliorated viability, and increased chronological life span. In an autophagy deficient background strain (atg1Δ), intracellular Aβ levels were not affected by UBB+1 expression. Our findings offer insights for reducing intracellular Aβ toxicity via autophagy-dependent cellular pathways under low level of UBB+1 expression.

Expression and content of VHL in kidney cancer tissue, relationship with clinical and morphological parameters of the disease, expression of transcriptional and growth factors

Introduction. The molecular picture of the development of HIF overexpression against VHL deficiency background is a key event associated with the manifestation and progression of clear cell renal cell carcinoma. However, this indicator's significance for the development of the disease and the formation of a response to targeted therapy is not clear. The study aimed to study the relationship between the expression and content of VHL in kidney cancer tissue with clinical, morphological parameters, mRNA level NF-κB p65, NF-κB p50, HIF-1, HIF-2, VEGF, and CAIX. Material and methods. The study included 39 patients with renal cell carcinoma. The disease's localized form (T1-2N0M0) was diagnosed in 20 patients; 19 patients had a disseminated process (T1-3N0-1M1). The study's material was normal, and tumor tissue was obtained during the surgical stage of treatment. VHL expression was determined by real-time PCR. The pVHL protein content was estimated using the Western Blotting method. Results. As a result of the study, it was noted that an increase in the VHL expression, together with a decrease in its protein level, was associated with an increase in tumor size. The prevalence of the disease was accompanied by an increase in both mRNA and VHL content in cancers. There was a negative correlation between the level of VHL expression, the content of the corresponding protein, and a positive association with the expression of NF-κB p65. VHL content in cancers was associated with decreased expression of NF-κB p50, NF-κB p65, and VEGF. Discussion. An increase in VHL expression in tumor tissue promoted the growth of mRNA of transcription factors, in particular NF-κB, which is accompanied by tumor spread. An increase in the VHL protein content led to the suppression of the mRNA level of transcriptional and growth factors, which is probably associated with their inactivation, including through proteasome cleavage, and is an important stage in oncogenesis. Conclusion. The relationship between the expression and content of the VHL protein is of decisive importance in the development of the disease, the formation of distant metastases.

Influence of androgen deficiency on the endometrium structure in women of reproductive age

Aim — to identify the relationship between androgen deficiency and the development of endometrial hypoplasia in women of reproductive age, to develop an algorithm for the diagnosis and treatment of this category of women. Materials and methods. Examination of patients with androgen deficiency revealed 48 patients with endometrial hypoplasia based on the ultrasound markers. After examination for CD138 and detection of chronic endometritis during the study, 9 patients were excluded. At the second stage, an immunohistochemical examination was performed for the expression of receptors for estrogen, progesterone and androgens. According to the results, the patients were divided into 2 groups: the first group (24 patients) with a high level of expression of androgen receptors and the second group (15 patients) with a low level of expression of androgen receptors. Theexpressionof receptors to estrogens and progesterone was on medium level and comparative in both groups. Both groups of patients underwent hormonal therapy for 3 months: estradiol valerate 1 g per day in a continuous mode and 200 mg of micronized progesterone from the sixteenth to twenty-fifth days of the menstrual cycle. Additionally, patients of the first group received dehydro­epian­drosterone(DHEA)in a dose of 25 mg per day continuously in the form sublingual spray. Results. According to the data of ultrasound examination in the first group of patients, the endometrium corresponded to normal parameters both during treatment and 1 and 3 months after stopping treatment. At the same time, in the second group of patients, there was an improvement in the thickness (more than 7 mm) and structure of the endometrium during treatment and the absence of these effects after the termination of hormonal therapy. Considering the recommendations of the Association of Endocrinologists on the superiority of non-tablet forms of androgen preparations in the treatment of androgen deficiency and having a positive and long-term effect when taking sublingual DHEA, it is possible to recommend adding the above form of DHEA to systemic therapy of endometrial hypoplasia against the background of androgen deficiency. Conclusions. Women with androgen deficiency are more likely to have concomitant endometrial hypoplasia. Immunohistochemical examination of the endometrium of women of reproductive age with androgen deficiency in 24 patients (61.5 %) revealed a high level of expression of androgen receptors. The effectiveness of therapy for endometrial hypoplasia in women with androgen deficiency with addition of androgens to the standard regimens is more effective and has a long-lasting effect. The combination of estrogen-gestagenic therapy and androgens has a positive effect on the gestational potential of the endometrium in women of reproductive age with androgen deficiency.

Co-production of hydrogen and ethyl acetate in Escherichia coli

BackgroundEthyl acetate (C4H8O2) and hydrogen (H2) are industrially relevant compounds that preferably are produced via sustainable, non-petrochemical production processes. Both compounds are volatile and can be produced by Escherichia coli before. However, relatively low yields for hydrogen are obtained and a mix of by-products renders the sole production of hydrogen by micro-organisms unfeasible. High yields for ethyl acetate have been achieved, but accumulation of formate remained an undesired but inevitable obstacle. Coupling ethyl acetate production to the conversion of formate into H2 may offer an interesting solution to both drawbacks. Ethyl acetate production requires equimolar amounts of ethanol and acetyl-CoA, which enables a redox neutral fermentation, without the need for production of by-products, other than hydrogen and CO2.ResultsWe engineered Escherichia coli towards improved conversion of formate into H2 and CO2 by inactivating the formate hydrogen lyase repressor (hycA), both uptake hydrogenases (hyaAB, hybBC) and/or overexpressing the hydrogen formate lyase activator (fhlA), in an acetate kinase (ackA) and lactate dehydrogenase (ldhA)-deficient background strain. Initially 10 strains, with increasing number of modifications were evaluated in anaerobic serum bottles with respect to growth. Four reference strains ΔldhAΔackA, ΔldhAΔackA p3-fhlA, ΔldhAΔackAΔhycAΔhyaABΔhybBC and ΔldhAΔackAΔhycAΔhyaABΔhybBC p3-fhlA were further equipped with a plasmid carrying the heterologous ethanol acyltransferase (Eat1) from Wickerhamomyces anomalus and analyzed with respect to their ethyl acetate and hydrogen co-production capacity. Anaerobic co-production of hydrogen and ethyl acetate via Eat1 was achieved in 1.5-L pH-controlled bioreactors. The cultivation was performed at 30 °C in modified M9 medium with glucose as the sole carbon source. Anaerobic conditions and gas stripping were established by supplying N2 gas.ConclusionsWe showed that the engineered strains co-produced ethyl acetate and hydrogen to yields exceeding 70% of the pathway maximum for ethyl acetate and hydrogen, and propose in situ product removal via gas stripping as efficient technique to isolate the products of interest.

Modern principles of diagnosis and treatment of severe traumatic brain injury

Traumatic brain injury (TBI) is a heterogeneous syndrome that includes various types of trauma. About 10–15 % of patients have severe TBI, accompanied by high rates of disability and mortality. Currently, the methods of intensive care from the standpoint of evidence-based medicine are widely used for their treatment. Along with intensive care, the comprehensive cytoprotective program using Neurocitin®, which contains citicoline and electrolytes, is of great importance. Neurocitin® has a combined effect aimed at correcting metabolic and volemic disorders, namely, maintaining adequate cerebral perfusion, timely elimination of water-electrolyte disorders, energy supply of brain tissues and protection of the neurovascular unit from damage. Lactate in the composition of Neurocitin® provides a detoxification effect, preserves and increases the energy supply of neurons, improves metabolism, cerebral blood flow and neuronal activity. For the treatment of TBI, the neurometabolic drug of ethylmethylhydroxypyridine succinate (Lodixem®) is also used. Lodixem® normalizes cerebral blood flow, provides an earlier resolution of post-traumatic encephalopathy and restoration of consciousness, leads to a decrease in the severity of endogenous intoxication syndrome, the elimination of pathological vicious circles that arise against the background of hypoxia, ischemia and energy deficiency in various organs and tissues.

Naphthalene Poisoning During a Decade: A Retrospective Clinical Study

: In this study, we investigated the naphthalene poisoning cases using the archived medical records of patients who were referred to Loghman-Hakim Hospital in Tehran, Iran, from 2008 to 2018. In the 52 studied cases, the highest prevalence was observed in the age groups of below five years and 20 - 25 years. Eighteen (34%) patients had no symptoms, and in others, gastrointestinal problems were the most common complaints. None of the patients needed intubation, and no one was admitted to the intensive care unit. Hemolysis was observed only in one patient with a background of G6PD deficiency. The average length of hospital stay was one day, and all patients improved without any complications. In general, naphthalene poisoning is not highly prevalent in Iran and usually does not cause severe morbidity and mortality. However, it is an available substance and can cause serious complications, especially in children. Therefore, it is necessary to investigate the clinical characteristics of these patients to improve the quality of health care.

A саsе of effective treatment of acne in patient with secondary immune deficiency of the first degree

Acne is an urgent problem in modern dermatology. The management of this disease is a complex, lengthy and multi-step process that requires a combination of different treatment methods and an individualized approach.
 Objeсtive — to demonstrate a clinical case of effective treatment of acne in a patient with secondary immune deficiency of the first degree.
 Materials and methods. A clinical case is presented of treatment of a 23-­year­-old man suffering from papulopustular form of moderate acne developed on the background of secondary immune deficiency of the first degree. The disease had a chronic course and resistance to the therapy used. The retinoid drug isotretinoin was used for treatment with a gradual increase and then decrease in its daily dose; antibiotic therapy; the drug Propes as an immunomodulator; as well as a set of cosmetic measures — chemical peelings and laser skin resurfacing. After the onset of dryness of certain areas of the skin, as a side effect of isotretinoin, emollients were locally intended.
 Results and discussion. At the end of the seventh month of treatment, the inflammatory rash on the skin completely stopped. The phenomenon of postacne was eliminated. During the control immunogram 2 months after the start of complex therapy, there was complete normalization of its parameters. In addition, during the first month of taking the drug isotretinoin , the patient did not have the exacerbation of the inflammatory process on the skin which was usual for this stage of treatment.
 Conclusions. The success in the treatment of the patient with acne who had secondary immune deficiency of the first degree as concomitant pathology, in our opinion, is due to a well­chosen comprehensive treatment which included antibacterial therapy, systemic retinoid isotretinoin in an appropriate for this clinical stage dose, Propes medication of the defensins group as an immunomodulatory agent, as well as the use of a complex of cosmetic measures.

Особливості перебігу бронхообструктивного синдрому у дітей раннього віку залежно від рівня забезпеченості вітаміном D

Актуальність. Захворювання органів дихання, і в тому числі бронхообструктивний синдром (БОС), є досить частою патологією дитячого віку. Відомо, що частота і тяжкість респіраторних захворювань корелюють із рівнем забезпеченості вітаміном D (VD), дефіцит якого може сприяти розвитку повторних епізодів БОС у дітей раннього віку поряд з іншими факторами ризику, такими як еозинофілія, обтяжений сімейний та індивідуальний алергоанамнез. Мета. Вивчення рівня забезпеченості VD і прогностичних критеріїв розвитку повторних епізодів БОС у дітей раннього віку. Матеріали та методи. Обстежено 120 дітей віком від 6 місяців до 3 років зі встановленим діагнозом «гострий обструктивний бронхіт» (1-ша група — 60 дітей з епізодичним БОС, 2-га група — 60 дітей із повторним БОС). Усім дітям проводили оцінку анамнестичних даних, загальне клінічне обстеження з визначенням ступеня тяжкості БОС за шкалою The Seattle Children’s respiratory scoring tool. Лабораторні дослідження включали визначення концентрації 25-гідроксивітаміну D (25(OH)D) в сироватці крові електрохемілюмінесцентним методом на аналізаторі Cobas e411 (серійний номер 1041-24, виробник Roche Diagnostics GmbH, Німеччина) та сироваткової концентрації загального кальцію (Са) за загальнопри­йнятою методикою. При аналізі отриманих даних використовували непараметричні статистичні критерії. Різниця між показниками, що порівнювались, вважалась вірогідною при р < 0,05. Результати. У дітей із повторним БОС на відміну від дітей з епізодичним БОС був значно нижчий рівень саплементації VD (р < 0,01). Перебіг БОС у дітей 2-ї групи вірогідно частіше супроводжувався розвитком синдрому дихальної недостатності порівняно з 1-ю групою (p < 0,01). За наявності повторних епізодів БОС в анамнезі захворювання характеризувалось більш тяжким перебігом. Середній показник 25(OH)D в сироватці крові у дітей 1-ї групи становив 33 (28,1; 41,9) нг/мл, 2-ї групи — 13,68 (7,96; 19,51) нг/мл (U = 152,0; p < 0,01). Достатній рівень забезпеченості VD частіше реєструвався у дітей 1-ї групи, тоді як у 2-й групі переважали діти, які мали дефіцит VD. Виявлено зворотну залежність між рівнем VD і частотою БОС. Рівень Са в сироватці крові в обох групах знаходився в межах нормальних величин. Висновки. Розвиток повторних епізодів БОС асоційований із більш низькими показники 25(OH)D у сироватці крові. Тяжкий перебіг БОС частіше зустрічається у дітей із повторними епізодами та на тлі дефіциту VD, який, однак, не супроводжується порушеннями кальцієвого обміну. Адекватний рівень забезпеченості VD найбільшою мірою забезпечується за рахунок достатнього рівня його саплементації, особливо на першому році життя. Таким чином, перебіг та частота епізодів БОС у дітей раннього віку залежать від рівня забезпеченості VD.