Decrease In Number Of Cells Research Articles

Intravenous haloperidol and cocaine alter the distribution of T CD3+ CD4+ , non-T/NK and NKT cells in rats.

The modulation of dopamine transmission evokes strong behavioural effects that can be achieved by commonly used psychoactive drugs such as haloperidol or cocaine. Cocaine non-specifically increases dopamine transmission by blocking dopamine active transporter (DAT) and evokes behavioural arousal, whereas haloperidol is a non-specific D2-like dopamine receptor antagonist with sedative effects. Interestingly, dopamine has been found to affect immune cells in addition to its action in the central nervous system. Here, we address the possible interactions between haloperidol and cocaine and their effects on both immune cells and behaviour in freely moving rats. We use an intravenous model of haloperidol and binge cocaine administration to evaluate the drugs' impact on the distribution of lymphocyte subsets in both the peripheral blood and the spleen. We assess the drugs' behavioural effects by measuring locomotor activity. Cocaine evoked a pronounced locomotor response and stereotypic behaviours, both of which were completely blocked after pretreatment with haloperidol. The results suggest that blood lymphopenia, which was induced by haloperidol and cocaine (except for natural killer T cells), is independent of D2-like dopaminergic activity and most likely results from the massive secretion of corticosterone. Haloperidol pretreatment prevented the cocaine-induced decrease in NKT cell numbers. Moreover, the increased systemic D2-like dopaminergic activity after cocaine administration is a significant factor in retaining T CD3+ CD4+ lymphocytes and non-T/NK CD45RA+ cells in the spleen.

The possible effect of lycopene in ameliorating experimentally induced ulcerative colitis in adult male albino rats (A histological, immunohistochemical, and ultrastructural study)

ABSTRACT Ulcerative colitis (UC) is considered a long-term inflammatory disorder worldwide. Its pathogenesis is associated with reduced antioxidant capacity. Lycopene (LYC) is a powerful antioxidant with strong free radical scavenging property. The present work has done to assess changes of colonic mucosa in induced UC and the possible ameliorative effects of LYC. Forty-five adult male albino rats were randomly divided into four groups: group I (control), group II was given 5 mg/kg/day (LYC) by oral gavage for 3 weeks. Group III (UC) was received single intra-rectal injection of acetic acid. Group IV (LYC+UC) received LYC in same dose and duration as before and acetic acid on 14th day of the experiment. UC group showed loss of surface epithelium with destructed crypts. Congested blood vessels with heavy cellular infiltration were observed. Significant decrease in goblet cell numbers and the mean area percentage of ZO-1 immunoexpression were noticed. Significant increase in the mean area percentage of collagen and the mean area percentage of COX-2 were also noticed. Ultrastructural changes were matched with light microscopic results that showed abnormal destructive columnar and goblet cells. Histological, immunohistochemical, and ultrastructural findings in group IV supported the ameliorative role of LYC against destructive changes induced by UC.

Secular decrease in chromosomal mosaicism in cultured and uncultured peripheral blood cells of six patients with mosaic Down syndrome.

As noted in a review paper,1 a large-scale study revealed that approximately 1.1%–3.8% of all children with Down syndrome (DS) are born with mosaic DS.2, 3 DS is diagnosed postnatally by chromosome G-banding analysis of peripheral blood cultures, and most patients do not undergo subsequent repeat analysis. We present secular changes in the mosaic ratio of six patients with mosaic DS. The G-banding, chromosome 21 fluorescence in situ hybridization (chr21 FISH) from peripheral blood cultures and the chr21 FISH analysis of uncultured blood cells showed a drastic reduction in the mosaic ratio in five patients aged between 7 and 23 years. The mosaic ratio of a child aged 3 years and 3 months gradually, but not drastically, decreased over time. However, no decrease in the mosaic ratio was seen in the buccal mucosa or fibroblasts by chr21 FISH in any mosaic patient. Low-invasive chromosomal analyses of 100 cells per patient from different tissues were performed by SRL, Inc. The analyses were as follows (Table 1). (1) T lymphocyte chromosome analyses of cultured peripheral blood samples by PHA (phytohemagglutinin) stimulation, including (1-a) G-banding and (1-b) chr21 FISH analysis using chromosome 21q22.13-q22.2 probes (Vysis, Abbott Molecular); (2) chr21 FISH analysis of uncultured blood cells, including various leukocytes, instead of bone marrow puncture; (3) chr21 FISH analysis of epithelial cells from the buccal mucosa; and (4) chr21 FISH analysis of fibroblasts cultured from a tooth extraction. The six patients (A–F) with mosaic DS (five females and one male) were aged from 3 years and 3 months to 23 years (Table 1). Five patients (B–F) aged between 7 and 23 years showed drastically decreased mosaic ratios (1%–8%) by G-banding, and FISH analysis of the peripheral blood culture and of uncultured blood cells compared with the postnatal result (Table 1, the red font). Patient A showed a gradual decrease in mosaicism on G-banding and FISH of the peripheral blood culture and by FISH analysis of uncultured blood cells at 11 months, 2 years and 9 months, and 3 years and 3 months of age compared with the postnatal result (Table 1, the red font). However, none of the six patients showed a decreased mosaic ratio in the buccal mucosa analysis or tooth-extraction-derived-fibroblast FISH analysis. Previously, two reports documented a decreased mosaic ratio in mosaic DS patients during early childhood, mainly in peripheral blood cultures.4, 5 We studied longitudinal changes in the mosaic ratio of six mosaic DS patients, including young adults. No decrease was observed in the epithelial cells of the buccal mucosa or cultured fibroblasts. The observed secular decrease in the mosaic ratio in cultured and uncultured peripheral blood cells did not appear to be a mechanism of trisomy rescue because six adult patients with standard trisomy 21 and one patient with translocated trisomy 21 showed no decrease in trisomy cell number in cultured and uncultured peripheral blood cells over time (Table S1, the green font). Within the bone marrow, hematopoietic stem cells exist in a hypoxic microenvironment (niche) and are in a quiescent (G0), undifferentiated state.6 In this microenvironment, trisomy cells are considered more detrimental to viability than normal cells, which may explain the secular decrease in chromosomal mosaicism with mosaic DS in this study. Permission has been obtained from the patients' mothers for the participation in this study. Financial support was provided by JSPS KAKENHI (Grant number: JP18K02497). The authors declare no conflict of interest. This study has received ethical approval from Tokyo Kasei University Ethics Committee (Ita h-29-5) and the Tokyo Metropolitan Tobu Medical Center for Children with Developmental Disabilities Ethics Committee (29MoriTobuCe256-1). TABLE S1. Chromosomal analyses of the trisomy ratio of eight patients, seven with standard trisomy 21 and one translocated trisomy 21 (patient H). Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Design, synthesis, and anti-triple negative breast cancer activity of novel Toosendanin derivatives

Toosendanin (TSN) is a natural anti-cancer compound that is isolated from the traditional Chinese herbal Melia toosendan Sieb et Zucc. However, the research effect of TSN in the treatment of Triple negative breast cancer (TNBC) is still far from ideal. In this work, we investigated TSN and its derivatives in terms of their actions against MDA-MB-231 and HCC1806 TNBC cell lines. The results indicated that TSN and its derivative 11 showed excellent antitumor activity. Preliminary mechanistic studies showed that both compounds TSN and 11 induced S-phase arrest and G2/M phase cell number decrease in HCC1806 cells. Also, TSN and 11 significantly reduced the protein level of the well-known cancer suppressor gene p53, reduced the phosphorylation of AKT and ERK, and also induced the accumulation of phosphorylated p38 and p21.

Nonylphenol reduced the number of haploids in in vitro spermatogenesis of the endangered cyprinid Gnathopogon caerulescens

Nonylphenol (NP), an endocrine disrupting chemical, is widely used in industrial and agricultural processes, causing NP influx into aquatic environments. NP induces hormonal imbalance, and male feminization, and reduces germ cell production during spermatogenesis; however, the mechanism by which it affects spermatogenesis remains unknown. Here, we investigated the effect of NP on spermatogenesis in honmoroko (Gnathopogon caerulescens), an endangered fish endemic to Lake Biwa, Japan, using an in vitro differentiation system. We collected spermatogonia from the testes of non-spawning G. caerulescens and subjected them to suspension culture. The spermatogonia differentiated into flagellated spermatozoa in 3 weeks, regardless of the presence of NP. NP concentrations as low as 1 nM caused a decrease in the number of germ cells in a dose-dependent manner, whereas the number of somatic cells decreased only at a high concentration of 1 μM. Flow cytometric analysis revealed that the decrease in germ cell number was attributed to haploids (spermatids and spermatozoa); the number of spermatogonia and spermatocytes was not affected by NP treatment. This result is consistent with the hypothesis that NP might repress the second meiosis or induce apoptosis in haploids. This study demonstrated that the combination of in vitro germ cell differentiation and flow cytometric analysis is useful for evaluating the direct effects of NP on germ cell differentiation in endangered endemic fish.

Biosynthesized silver nanoparticles have anticoccidial and jejunum-protective effects in mice infected with Eimeria papillata.

Eimeriosis, an infection with Eimeria spp. that affects poultry, causes huge economic losses. Silver nanoparticles (AgNPs) have antibacterial and antifungal properties, but their action against Eimeria infection has not yet been elucidated. This study demonstrates the action of AgNPs in the treatment of mice infected with Eimeria papillata. AgNPs were prepared from Zingiber officinale rhizomes. Phytochemical screening by gas chromatography-mass spectrometry analysis (GC-MS) was used to detect active compounds. Mice were divided into five groups: uninfected mice, uninfected mice that were administered AgNPs, untreated mice infected with 103 sporulated oocysts of E. papillata, infected mice treated with AgNPs, and infected mice treated with amprolium. Characterization of the samples showed the AgNPs to have nanoscale sizes and aspherical shape. Phytochemical screening by GC-MS demonstrated the presence of 38 phytochemical compounds in the extract of Z. officinale. Mice infected with E. papillata-sporulated oocysts were observed to have many histopathological damages in the jejuna, including a decrease in the goblet cell numbers affecting the jejunal mucosa. Additionally, an increased oocyst output was also observed. The treatment of infected mice with AgNPs resulted in the improvement of the jejunal mucosa, increase in the number of goblet cell, and decrease in the number of meronts, gamonts, and developing oocysts in the jejuna. Moreover, AgNPs also led to decreased oocyst shedding in feces. The results revealed AgNPs to have an anticoccidial effect in the jejunum of E. papillata-infected mice and, thus, could be a potential treatment for eimeriosis.

Glucocorticoid-sensitive period of corticotroph development-Implications for mechanisms of early life stress.

Corticotrophs are intermediaries in the hypothalamic-pituitary-adrenal (HPA) axis, which plays a crucial role in stress response in vertebrates. The HPA axis displays an intricate mode of negative feedback regulation, whereby the peripheral effector, cortisol inhibits the secretion of its upstream regulator, adrenocorticotropic hormone (ACTH) from proopiomelanocortin (POMC)-expressing cells in the pituitary. While the feedback regulation of the HPA axis is well characterized in the adult organism, the effect of feedback regulation on the development of corticotrophs is poorly understood. Here, we studied the effect of glucocorticoids on the development of POMC-expressing cells in the zebrafish pituitary. The development of POMC cells showed a steady increase in numbers between 2-6 days post fertilization. Inhibition of endogenous glucocorticoid synthesis resulted in an increase in POMC cell number due to reduced developmental feedback inhibition of cortisol on POMC cells. Conversely, addition of exogenous dexamethasone at a critical developmental window led to a decrease in corticotroph cell number, mimicking greater feedback control due to increased cortisol levels. Finally, developmental dysregulation of ACTH levels resulted in impaired anxiety-like and stress-coping behaviours. Hence, we identified a sensitive developmental window for the effect of glucocorticoids on corticotrophs and demonstrate the downstream effect on stress-responsive behaviour.

The Role of Purine Metabolism-Related Genes PPAT and IMPDH1 in the Carcinogenesis of Intrahepatic Cholangiocarcinoma Based on Metabonomic and Bioinformatic Analyses.

In this study, we investigated the role of tumor microenvironment and serum differential metabolites in intrahepatic cholangiocarcinoma (ICC) carcinogenesis, providing new evidence for ICC treatment. Serum samples from healthy individuals and ICC patients were collected for metabolomic analysis. The purine metabolites such as inosine, guanosine, hypoxanthine, and xanthine were increased in patient serum. TCGA database samples were collected, and the correlation between purine metabolism-related genes and ICC clinical features was analyzed using R language to obtain the differential genes including PPAT, PFAS, ATIC, and IMPDH2. High PPAT expression was associated with poor ICC prognosis. A PPAT silencing model in HCCC-9810 cells was constructed. The cell phenotype was examined by qRT-PCR, CCK-8, transwell, and flow cytometry, showing a decrease in IMPDH1 expression, colony and invasive cells numbers, and an increase in apoptosis. Guanosine reversed IMPDH1 expression in HCCC-9810 cells, promoting the secretion of inflammatory factors IL-6, IL-8, OPN, VEGF, and VCAM-1 and intensifying epithelial-mesenchymal transition (EMT) progression in the cells. In nude mice, the IMPDH1 inhibitory drug MMF inhibited tumor growth and reduced the expression of tumor stem cell characteristic markers CD133 and SOX2. Guanosine accelerated the malignant progression of ICC inhibition of purine metabolism-related genes, PPAT and IMPDH2, suppressed the malignant phenotype in HCCC-9810 cells, and inhibited tumor growth.

23-Hydroxybetulinic Acid, A Natural Compound, Alleviates DSS-induced Colitis by Regulating NF-κB Signaling

Article 23-Hydroxybetulinic Acid, A Natural Compound, Alleviates DSS-induced Colitis by Regulating NF-κB Signaling Shuangli Xiang 1, # , Miaojuan Wang 2, # , and Xiuping Chen 2, * 1 Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi, Guizhou Province, China. 2 State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macao, China. * Correspondence: xpchen@um.edu.mo, Tel.: +853-88224679, Fax: +853-28841358 # Co-First author. Received: 8 November 2022 Accepted: 2 December 2022 Published: 11 January 2023 Abstract: Ulcerative colitis (UC), an inflammatory intestinal disease, is a growing epidemic affecting people worldwide and requires the development of effective therapeutic drugs. In this study, the effect of 23-hydroxybetulinic acid (23-HBA), a compound isolated from the traditional herb Pulsatilla chinensis (Bunge) Regel, on experimental UC was studied. C57BL/6J male mice were administrated with 3% dextran sodium sulfate (DSS) in drinking water to establish the UC model. 23-HBA was orally administrated at either 3.75, 7.5, or 15 mg/kg for 6 days. Mesalazine was used as a positive control. Examination of the body weight, colon length, disease activity index (DAI), histopathology examination, inflammatory cytokines, oxidative stress, and protein expression was performed. The pathological changes were examined with hematoxylin and eosin (H&E) and Aixian blue-glycogen (AB-PAS) staining. In cultured RAW 264.7 cells, the effects of 23-HBA on lipopolysaccharide (LPS)-stimulated cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and oxidative stress were analyzed. Compared with the colitis model, 23-HBA treatment significantly increased the body weight and colon length and decreased the DAI score. Pathological staining showed that 23-HBA mitigated the damage in intestinal structures, the increase in inflammatory cell infiltration, the increase in submucosa edema, and the decrease in goblet cell number. Furthermore, 23-HBA decreased IL-1β, IL-6, and MDA levels in the colon tissues. In addition, 23-HBA inhibited the protein expressions of COX-2, iNOS, and NF-κB p65 both in the colon tissues and in LPS-stimulated RAW 264.7 cells. In conclusion, these results showed that 23-HBA alleviated DSS-induced acute UC in mice and inhibited LPS-stimulated inflammation in RAW 264.7 cells possibly mediated by regulating the NF-κB pathway.

Effects of thyroid hormones on the functional state of bovine granulosa cells in vitro

Any dysfunction of the thyroid gland causes abnormal changes in the functioning of the reproductive system, primarily the ovaries. Therefore, the question of the possible direct effect of thyroid hormones on the bovine ovarian function by modulating the functional state or functional activity of granulosa cells seems relevantPurpose: to study in vitro the effect of thyroxine and triiodothyronine on the proliferative and steroidogenic activity, as well as apoptotic changes of bovine granulosa cells.Materials and methods. Granulosa cells were isolated from follicles with a diameter of 1-5 mm and precultured for two days in a medium containing 10 % serum. The cells were then placed in serum-free medium containing thyroxine (25-200 ng/mL) or triiodothyronine (0.5-4.0 ng/mL) and incubated for another 48 h. After culturing, the content of estradiol-17β and progesterone in the media was determined by ELISA. The proliferative activity and apoptotic changes in the cells were assessed by immunocytochemical assay, based on the expression level of proliferating cell nuclear antigen PCNA and pro-apoptotic protein Bax, respectively.Results. It was found that the proportion of cells with a positive reaction to PCNA increased 1.1 times (P<0.01) compared with that in the control at a triiodothyronine concentration of 1 ng/ml and did not change with its further increase to 4 ng/ml. In addition, the introduction of triiodothyronine at a concentration of 1 ng/ml into the medium led to a decrease in the relative number of Bax-positive cells from 25.6 ± 0.3% to 23.3 ± 0.6 % (P<0.01). A further increase in this concentration to 4 ng/ml enhanced the observed anti-apoptotic effect 1.1 times (P<0.05). The pattern of the effect of thyroxine on the proliferative activity and apoptotic changes of granulosa cells in culture was similar to that for triiodothyronine. Concurrently, the growth-stimulating and anti-apoptotic effects of thyroxine were achieved at a concentration of 50-200 ng/ml. At the same time, both thyroid hormones did not affect the secretion of estradiol-17β or progesterone by the cells.Conclusions. Thus, thyroxine and triiodothyronine can stimulate in vitro the proliferation of bovine granulosa cells, as well as inhibit the expression of the proapoptotic Bax protein in these cells, which is not associated with the regulation of the production of ovarian steroid hormones. Overall, these data suggest that thyroid hormones at physiological concentrations are able to exert a regulatory effect on the growth and atresia of bovine small antral follicles and, therefore, directly modulate the ovarian activity.

Management of compensatory shift of immunohematological parameters in poultry of industrial crosses in the presence of feed antibiotics in the diets

A comparative analysis of the immunological parameters in chickens of industrial crosses, which were fed diets with antibiotics (group 1) and antibiotics in combination with a phytobiotic (group 2), is presented. After 30 days of feeding antibiotic hemoglobin concentration in the blood of laying hens remained unchanged – 87.3-93.2 g/l. On the 30th day of the experiment, in group 1 was noted an increase in the number of leukocytes up to 30.13±2.84∙109/l and the proportion of eosinophils by 29.6%. In the same period, a decrease in the absolute number of immunocompetent cells – T-lymphocytes by 36.6% was observed and absorption capacity of phagocytes by an average of 27.9%. From the 30th to the 60th day of the experiment in the blood of the chickens of group 1, the dynamics of the increase in the CIC level was recorded (up to 143.8±12.1 c.u.). When antibiotic was introduced into the diet of laying hens in combination with a phytobiotic in group 2, only by the 60th day, the number of erythrocytes was increased by 17.4% compared to the “background values” (82.1±5.2∙1012/l). Hemoglobin fluctuation amplitude did not exceed 7%. On the 30th day of the experiment in laying hens of group 2, a decrease in the number of immunocompetent cells by 20.8% was registered compared to the background values. On the 45th day of the experiment after the abolition of antibiotic and phytobiotic, the content of T-lymphocytes did not significantly differ from the background values.

Effects of Brimonidine on Microglia Morphology in the Transient Retinal Ischemia Model

Background:This study aimed to investigate the effects of brimonidine on microglia cell morphology by creatinga transient retinal ischemia model in rats. Methods:In the right eyes of male Wistar rats (n= 12), a transient retinal ischemia model was created. The ratswere divided into three groups: (1) eyes treated with topical brimonidine in the transient retinal ischemia model,(2) sham-treated eyes, and (3) control eyes. Four main phenotypes (ramified, primed, reactive, and amoeboid-phagocytic) of Iba-1 positive microglia cells were examined in the retinal layers. Results:In the transient retinal ischemia model, the number of Iba-1 positive microglia cells was 100.67±7.50 cellsin the sham group and 57.67±14.64 cells in the topical brimonidine group. The decrease in the total number of Iba-1 positive microglial cells was statistically significant (p<0.05). When we compared ramified and primed microgliacells, there was no statistically significant difference between the groups. The number of amoeboid-phagocyticmicroglial cells was 9.5±1.29 cells in the sham treatment group and 2.25±0.50 cells in the topical brimonidinegroup (p<0.05). There was a statistically significant decrease in the number of reactive and amoeboid cells. Conclusion:Studies in the central nervous system in rats demonstrated that cell metabolism and functions werechanged after acute injury and that microglia transformed from a ramified form to an amoeboid-phagocytic form.In this study, the decrease in the transition of cells from ramified to reactive and amoeboid forms showed us thattopical brimonidine treatment could have neuroinflammation suppressor features.

Morphology of the testes of animals during estrogen injection in the prenatal period

The aim of the study was to investigate the morphological changes in the testes of the offspring of white non-pedigreed laboratory mice when the synthetic oestrogen analogue synestrol was prenatally injected to the mother. After fertilization, the females were divided into 2 groups which received a single intramuscular injection of the synthetic oestrogen analogue synestrol on day E 11.5 of gestation at the same time of the day. The intact group was not exposed to any treatment. The experimental group was exposed to synestrol, a synthetic oestrogen analogue, as a 2% oil solution at a dose of 40 µg/kg (C-40). The results of morphometric analysis of testes of descendants of white outbred laboratory mice after prenatal single injection of synthetic oestrogen analogue synestrol in a dose of 40 µg/kg showed morphological changes in the organ parenchyma, manifested as decrease in mean number of Sertoli cells (C-40) 17.4±1.1 compared to intact group 20.8±1.9; decrease in mean number of spermatogonia (C-40) 24.4±1.1 compared to intact group, decrease in mean number of spermatozoa (C-40) 178.0±4.2 compared to intact group 196.6±5.3. There was observed a change in the endocrine apparatus of testes, expressed as a decrease in the mean area of Leydig cell nuclei in the intact group of 6,72±1,78. Prenatal effects of synestrol revealed in an experimental model, allow us to use it to search for means of postnatal developmental testicular dysfunction correction as well as to develop optimal doses of oestrogenic preparations during pregnancy.

HEMATOLOGICAL PARAMETERS IN THE MODEL OF IRON DEFICIENCY AND HEMATOLOGICAL ANEMIA

According to the World Health Organization, the most common (up to 90% of cases) hematological pathology is anemia, a condition characterized by a decrease in the level of hemoglobin in the blood due to a decrease in the total number of red blood cells or a violation of hemoglobin synthesis, which can have serious consequences for the body, such as decreased immunity, impaired respiratory and metabolic processes. Currently, it is necessary to improve the methods of prevention and treatment of anemia. To test new drugs for these purposes, it is necessary to create new models, including on laboratory animals. The aim of the study was to develop experimental models of iron deficiency (IDA) and hemolytic anemia (HA) on laboratory animals. The objects of the study were sexually mature male laboratory rats of the Wistar breed, aged 7 months and weighing 260 ± 20 g. 38 animals were involved in the experiment (for IDA n= 24, for HA n= 14). Laboratory animals were kept in the same external conditions for 3-4 individuals in a cage. To simulate IDA, rats were subcutaneously injected with the drug “Desferal”. To simulate GA, rats were intraperitoneally injected with the drug “Hydrochloric acid phenylhydrozine”. At each stage, blood was taken by cutting off the tip of the tail. A detailed blood test was performed on a hematological analyzer “Heska Element HT5”. As a result, it was found that when the drug “Desferal” was administered at concentrations of 125 and 250 mg / 100g per animal weight, hematological changes were observed in rats, such as an increase in red blood cells, platelets, leukocytes by stage 2 and a decrease to control values by stage 4. When the drug “Hydrochloric acid phenylhydrozine” was administered, hematological changes were detected in rats only within the experimental group. There was an increase in the indicators of erythrocytes, platelets, leukocytes by stages 2 and 3 and a decrease to control values by stage 4. Thus, when the studied drugs were administered in the presented dosages in vivo, a change in the hematological parameters of the blood was detected in the direction of an increase in their number due to adaptation to the effects of substances.

In Vitro Self-Assembly of a Modified Diphenylalanine Peptide to Nanofibers Induced by the Eye Absent Enzyme and Alkaline Phosphatase and Its Activity against Breast Cancer Cell Proliferation.

Drug-resistant breast cancers such as Triple negative breast cancer (TNBC) do not respond successfully to chemotherapy treatments because they lack the expression of receptor targets. Drug-resistant anti-cancer treatments require innovative approaches to target these cells without relying on the receptors. Intracellular self-assembly of small molecules induced by enzymes is a nanotechnology approach for inhibiting cancer cell growth. In this approach, enzymes will induce the self-assembly of small molecules to nanofibers, which leads to cell death. Here, we investigate the self-assembly of a modified small peptide induced by two different phosphatases: alkaline phosphatase (ALP) and eye absent tyrosine phosphatase (EYA). ALPs are expressed in many adult human tissues and are critical for many cellular functions. EYAs are embryonic enzymes that are over-expressed in drug-resistant breast cancers. We synthesized a small diphenylalanine-based peptide with a tyrosine phosphate end group as the substrate of phosphatase enzymes. Peptides were synthesized with solid phase techniques and were characterized by HPLC and MALDI-TOF. To characterize the self-assembly of peptides exposed to enzymes, different techniques were used such as scattering light intensity, microscopes, and phosphate detection kit. We then determined the toxicity effect of the peptide against normal breast cancer cells, MCF-7, and drug-resistant breast cancer cells, MDA-MB-231. The results showed that the EYA enzyme is able to initiate self-assembly at lower peptide concentration with higher self-assembling intensity compared to ALP. A significant decrease in the TNBC cell number was observed even with a low peptide concentration of 60 μM. These results collectively support the exploration of enzyme self-assembly to treat TNBC.

Sodium Butyrate Supplementation Modulates Neuroinflammatory Response Aggravated by Antibiotic Treatment in a Mouse Model of Binge-like Ethanol Drinking.

Growing evidence supports the pivotal role of the bidirectional interplay between the gut microbiota and the central nervous system during the progression of alcohol use disorder (AUD). In our previous study, supplementation with sodium butyrate (SB) in C57BL/6J mice prevented increased ethanol consumption in a binge-like drinking paradigm (DID) as a result of treatment with a non-absorbable antibiotic cocktail (ABX). In this study, we tested the hypothesis that SB protection against enhanced ABX-induced ethanol consumption in mice is partially due to modulation of neuroinflammatory responses. Pro- and anti-inflammatory cytokines, as well as changes in microglia and astrocytes were analyzed in hippocampus tissues from ABX-, SB-, ABX+SB-treated mice subjected to 4-week DID. We found that ethanol without or with ABX treatment increased mRNA levels of key brain cytokines (MCP-1, TNF-α, IL-1β, IL-6 and IL-10) while SB supplementation prevented these changes. Additionally, SB supplementation prevented changes in microglia, i.e., increase in Iba-1 positive cell number and morphology, and in astrocytes, i.e., decrease in GFAP-positive cell number, induced by combination of ethanol and ABX treatments. Our results suggest that gut microbiota metabolites can influence drinking behavior by modulation of neuroinflammation, highlighting the potential for microbiome-targeting strategies for treatment or prevention of AUD.

Subpopulations of lymphocytes and monocytes in blood of patients with atrial fibrillation or atrial flutter associated with hypertension

Atrial fibrillation (AF) remains one of the most common arrhythmias, second only to supraventricular extrasystoles, but the universal cause of its occurrence is still unknown. The inflammatory theory of arrhythmogenesis attracts the attention of researchers around the world. The purpose of the study is to compare the subpopulations of lymphocytes and monocytes in blood of patients with paroxysmal and persistent forms of atrial fibrillation or atrial flutter (AFL) that associated with arterial hypertension. Materials and methods. The study involved 103 patients with atrial fibrillation and flutter that occurred secondary to hypertension. Depending on the form of arrhythmia, they were divided into three main groups: group I – with paroxysmal form of atrial fibrillation, group II – with a persistent form of atrial fibrillation, group III – with a persistent form of atrial flutter. The control groups included patients with hypertension, but without these arrhythmias and healthy individuals who entered groups IV and V, respectively. The lymphocytes and monocytes subpopulation was assessed by flow cytometry in peripheral blood. Results and discussion. Analyzing the lymphocyte subpopulations in peripheral blood of patients with atrial fibrillation and flutter (groups I, II and III), it was found that the number of cells with cytotoxic activity (NK and NKT) in both absolute count and percentage values was significantly higher than in healthy individuals. A statistically significant decrease of T-regulatory cells number was found in patients with arrhythmias compared to control groups (p ≤ 0.05). In patients with AF and AFL associated with hypertension, compared to patients with hypertension without these rhythm disturbances or healthy individuals, there is an increased number of classical and intermediate monocytes subpopulations. Conclusions. In patients with atrial fibrillation and atrial flutter that occurred as a result of hypertension, compared to patients without arrhythmias or healthy people, there is an increased content of pro-inflammatory subpopulations of blood monocytes, T-cytotoxic cells and a decrease in the content of T-regulatory cells.

Syndecan-4 regulates the HER2-positive breast cancer cell proliferation cells via CK19/AKT signalling

Despite the use of the highly specific anti-HER2 receptor (trastuzumab) therapy, HER2-positive breast cancers account for 20–30% of all breast cancer carcinomas, with HER2 status a challenge to treatment interventions. The heparan sulfate proteoglycans (HSPGs) are prominently expressed in the extracellular matrix (ECM), mediate breast cancer proliferation, development, and metastasis with most studies to date conducted in animal models. This study examined HSPGs in HER2-positive human breast cancer cell lines and their contribution to cancer cell proliferation. The study examined the cells following enhancement (via the addition of heparin) and knockdown (KD; using short interfering RNA, siRNA) of HSPG core proteins. The interaction of HSPG core proteins and AKT signalling molecules was examined to identify any influence of this signalling pathway on cancer cell proliferation. Our findings illustrated the HSPG syndecan-4 (SDC4) core protein significantly regulates cell proliferation with increased BC cell proliferation following heparin addition to cultures and decreased cell number following SDC4 KD. In addition, along with SDC4, significant changes in CK19/AKT signalling were identified as mediators of BC HER2-positive BC cell proliferation. This study provides evidence for a cell growth regulatory axis involving HSPGs/CK19 and AKT that represents a potential molecular target to prevent proliferation of HER2-positive breast cancer cells.

Isolation, Characterization and Anticancer Activity of Two Bioactive Compounds from Arisaema flavum (Forssk.) Schott.

Medicinal plants play important role in the public health sector worldwide. Natural products from medicinal plants are sources of unlimited opportunities for new drug leads because of their unique chemical diversity. Researchers have focused on exploring herbal products as potential sources for the treatment of cancer, cardiac and infectious diseases. Arisaema flavum (Forssk.) is an important medicinal plant found in the northwest Himalayan regions of Pakistan. It is a poisonous plant and is used as a remedy against snake bites and scorpion stings. In this study, two bioactive compounds were isolated from Arisaema flavum (Forssk.) and their anticancer activity was evaluated against human breast cancer cell line MCF-7 using an MTT assay. The crude extract of Arisaema flavum (Forssk.) was subjected to fractionation using different organic solvents in increasing order of polarity. The fraction indicating maximum activity was then taken for isolation of bioactive compounds using various chromatographic and spectroscopic techniques such as column chromatography, thin-layer chromatography (TLC), gas chromatography-mass spectrometry (GC-MS), Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy (NMR). Crude extract of Arisaema flavum (Forssk.), as well as various fractions extracted in different solvents such as n-hexane, chloroform and ethyl acetate, were tested against human breast cancer cell line MCF-7 using an MTT assay. The crude extract exhibited significant dose-dependent anticancer activity with a maximum activity of 78.6% at 500 µg/mL concentration. Two compounds, hexadecanoic acid ethyl ester with molecular formula C18H36O7 and molar mass 284 and 5-Oxo-19 propyl-docosanoic acid methyl ester with molecular formula C26H50O3 and molecular mass 410, were isolated from chloroform fraction. These compounds were tested against the MCF-7cell line for cytotoxic activity and exhibited a significant (p < 0.00l) decrease in cell numbers for MCF-7 cells with IC50 of 25 µM after 48 h of treatment. Results indicated that Arisaema flavum (Forssk.) possesses compounds with cytotoxic activity that can further be exploited to develop anticancer formulations.

ClNAC84 interacts with ClMIP to regulate the cell cycle and reduce the size of Chrysanthemum lavandulifolium organs

The NAC transcription factor is plant-specific proteins and one of the largest families of transcription factors in plants. NAC proteins are involved in various aspects of plant growth and development, but little is known about how NAC proteins regulate the cell cycle. Here, we characterized ClNAC84 from C. lavandulifolium (an NAC transcription factor). ClNAC84 overexpression in C. lavandulifolium resulted in a semi-dwarf phenotype with shorter plant height, smaller leaf size, and smaller flower size than wild-type plants. The number of cells in the S phase during the cell cycle was less in ClNAC84-overexpression transgenic C. lavandulifolium than in wild-type C. lavandulifolium. This indicates that ClNAC84 overexpression can induce cell cycle arrest at the S and G2 phases. To elucidate the ClNAC84 regulatory network, ClMIP protein was shown to interact with ClNAC84 in vitro and in vivo. ClMIP overexpression in C. lavandulifolium also resulted in dwarfism and decreased cell numbers, and the expression level of ClKRP5 was higher in transgenic C. lavandulifolium than in wild-type plants. We also found that ClMIP can bind to the promoter of ClKRP5. Our data indicate that the interaction between ClNAC84 and ClMIP may promote ClKRP5 expression and inhibit S and G2 phases of the cell cycling.