Cell Atrophy Research Articles

Growth, Biochemical Characteristics, Flesh Quality, and Gut Microbiota of the Pacific White Shrimp (Penaeus vannamei) Fed a Defatted Superworm (Zophobas atratus) Larvae Meal.

This study evaluated the effects of defatted superworm (Zophobas atratus) larvae meal (DBWLM) as an alternative protein ingredient for juvenile Pacific white shrimp (Penaeus vannamei). Six isonitrogenous and isolipidic experimental diets were characterized by replacing 0%, 15%, 30%, 45%, 60%, and 75% fish meal (DBWLM0, DBWLM15, DBWLM30, DBWLM45, DBWLM60, and DBWLM75, respectively) with DBWLM on a w/w basis and feeding them to juvenile shrimp (0.34 ± 0.04 g) for 56 days. The results showed that the replacement of up to 75% fish meal by DBWLM had no negative effect on the growth performance of P. vannamei. The survival of shrimp in the DBWLM30 group was the highest, and the weight gain, specific growth rate, feed conversion ratio, condition factor, and apparent digestibility coefficients of dry matter in the DBWLM15 group were the highest. The substitution of DBWLM for fish meal significantly increased the elasticity of flesh, improved the total content of umami amino acids in flesh (aspartic acid, glutamic acid, glycine, and alanine), promoted lipid metabolism in shrimp, and reduced serum lipid levels. With the increase in DBWLM level, serum acid phosphatase, alkaline phosphatase activity, and intestinal inflammatory gene expression (IGF-1 and IL-6) were inhibited, malondialdehyde content decreased, and total antioxidant capacity level and superoxide dismutase activity increased significantly. Histological sections of the hepatopancreas showed that when 60% or more fish meal was replaced, the hepatopancreas atrophied and had irregular lumen distortion, but the cell membrane was not damaged. Microbiome analysis showed that the abundance of Bacteroidetes and Firmicutes increased and the abundance of Proteobacteria decreased in the DBWLM replacement group, and it was rich in "metabolism"-related functional pathways. It is worth mentioning that the expression of amino-acid-related enzymes was upregulated in the DBWLM15 and DBWLM30 groups, and the DBWLM75 group inhibited the biosynthesis of steroids and hormones. To conclude, the replacement of 15%-45% fish meal with DBWLM can result in better growth and immune status, improved meat elasticity, and reduced inflammation in P. vannamei. However, it is recommended that the replacement level should not exceed 60%, otherwise it will cause atrophy of hepatopancreas cells.

P-030 Clinical outcomes of microdissection testicular sperm extraction (micro TESE) and intracytoplasmic sperm injection (ICSI) in non-obstructive azoospermia (NOA) with the history of cryptorchidism

Abstract Study question What is sperm retrieval rate (SRR) by micro TESE and the clinical outcomes using testicular sperm in couples with the history of cryptorchidism? Summary answer NOA couples with the history of cryptorchidism had a higher SRR by micro TESE but lower clinical outcomes by ICSI compared of unexplained NOA. What is known already Undescended testis (UT) which is exposed to a higher temperature compared with the scrotal temperature is associated with impairment of germ cell maturation, and progressive Leydig and Sertoli cell atrophy, and subsequent infertility in adulthood. There have been very few studies of ICSI with a focus on, or large enough numbers to examine, the specific outcomes associated with male factor infertility, however improvement in sperm retrieval techniques including micro TESE and micromanipulation techniques, such as ICSI, has led to excellent fertilization and pregnancy outcomes of treatment cycles. Study design, size, duration We performed a retrospective study based on two reproduction centers in Japan and evaluated 1521 azoospermic patients in our clinics between September 2013 and December 2022. We investigated SRR by micro TESE in these patients and therefore aimed to evaluate the prevalence and the significance of ICSI outcomes with embryonic development in NOA couples with the history of cryptorchidism. Participants/materials, setting, methods We evaluated SRR of micro TESE, two pronuclei (2PN) oocyte rates, blastocyst development, good-quality blastocyst (Grade 3BB and above on day 5 by the Gardner scoring), and clinical pregnancy rates per embryo transfer (ET) in 72 NOA cases with the history of UT, 953 cases of unexplained NOA, not including after orchidopexy, Klinefelter syndrome, cryptozoospermia, mumps orchitis, and 284 cases of obstructive azoospermia (OA). Statistical analysis was performed using unpaired t-tests and chi-squared tests. Main results and the role of chance SRR of first attempt micro TESE in UT (44/56=78.6%) was higher than unexplained NOA (160/748=21.4%) (p < 0.001). Spermatozoa were successfully retrieved in 13 of 22 (59.1%) UT group and 36 of 226 (15.9%) unexplained NOA who had previously undergone micro TESE with no sperm found. No correlation was found between serum FSH, LH, and T level with the success of sperm retrieval. Testicular volume and patient age at orchidopexy also did not affect the SRR for micro TESE. 2PN oocytes, blastocysts development, and good-quality blastocysts rates were 45.5%, 47.4%, and 18.9% in UT, 51.2%, 45.0%, and 20.9% in unexplained NOA, and 62.6%, 52.1%, and 23.7% in OA, respectively. Clinical pregnancy rates per ET were 29.3% in UT, 32.3% in unexplained NOA, and 41.6% in OA. Significant differences were only observed in 2PN oocytes between unexplained NOA and OA (p < 0.05) and clinical pregnancy between UT and unexplained NOA (p < 0.05), and OA and unexplained NOA (p < 0.01). Several UT patients showed a very small number of spermatozoa and even only immotile sperm of retrieved by micro TESE. Limitations, reasons for caution We included the patients only after a surgery of cryptorchidism, but not a delayed testicular descent without surgery. The cohort size of this study is not small, however, our screened population of azoospermic patients may be biased. Wider implications of the findings This study shows a high impact in micro TESE and ICSI outcomes with embryonic development for the NOA couples with the history of cryptorchidism. Our study emphasizes that history of cryptorchidism provides clinically valuable prognostic information to couples considering surgical sperm retrieval. Trial registration number not applicable

Growth inhibition, oxidative stress and characterisation of mortality in green algae under the influence of beta-blockers and non-steroidal anti-inflammatory drugs

Bisoprolol and ketoprofen are widely used pharmaceuticals in medical treatment hence these substances are occurring in wastewaters and in water environment. This research investigated the toxic effects of bisoprolol and ketoprofen on two microalgae taxa, Chlorella vulgaris and Desmodesmus armatus. The results showed that both drugs inhibited the growth of the species tested and induced a decrease in chlorophyll a content compared to controls. Ketoprofen turned out to be harmful to algae as the half maximal effective concentration (EC50) values (14 days) were 37.69 mg L−1 for C. vulgaris and 40.93 mg L−1 for D. armatus. On the other hand, for bisoprolol, the EC50 values were greater than the established NOEC, 100 mg L−1. Bisoprolol and ketoprofen induced oxidative stress in the tested microorganisms, as indicated by changes in the activities of antioxidant enzymes. Exposure to 100 mg L−1 of drugs significantly increased the activity of catalase, peroxidase and superoxide dismutase. Fluorescence microscopy showed that both medicaments changed the cells' morphology. There was atrophy of chlorophyll in the cells, moreover, dying multinuclear cells and cells without nuclei were observed. In addition, there were atrophic cells, namely cells that lacked nuclei and chlorophyll. Profile area analyses showed that bisoprolol and ketoprofen treated C. vulgaris cells were approximately 4 and 2 times greater compared to control ones. Our experimental findings highlight the ecotoxicological threats for aquatic primary producers from bisoprolol and ketoprofen and provide insight into the characteristics of their death.

Stimulated myotube contractions regulate membrane-bound and soluble TLR4 to prevent LPS-induced signaling and myotube atrophy in skeletal muscle cells.

Toll-like receptor 4 (TLR4) activation by lipopolysaccharides (LPS) increases pro-inflammatory cytokine production and upregulation of muscle atrophy signaling pathways. Muscle contractions can suppress LPS/TLR4 axis activation by reducing the protein expression of TLR4 on immune cells. However, the mechanism by which muscle contractions decreases TLR4 remains undefined. Moreover, it is not clear whether muscle contractions affect TLR4 expressed on skeletal muscle cells. The purpose of this study was to uncover the nature and mechanisms by which stimulated myotube contractions using electrical pulse stimulation (EPS) as an in vitro model of skeletal muscle contractions affect TLR4 expression and intracellular signaling to combat LPS-induced muscle atrophy. C2C12 myotubes were stimulated to contract via EPS with and without subsequent LPS exposure. We then examined the isolated effects of conditioned media (CM) collected following EPS and soluble TLR4 (sTLR4) alone on LPS-induced myotube atrophy. Exposure to LPS decreased membrane-bound and sTLR4, increased TLR4 signaling (decreased inhibitor of κBα), and induced myotube atrophy. However, EPS decreased membrane-bound TLR4, increased sTLR4, and prevented LPS-induced signaling and myotube atrophy. CM, that contained elevated levels of sTLR4, prevented LPS-induced upregulation of atrophy-related gene transcripts muscle ring finger 1 (MuRF1) and atrogin-1 and reduced myotube atrophy. Recombinant sTLR4 added to media prevented LPS-induced myotube atrophy. In summary, our study provides the first evidence that sTLR4 has anti-catabolic effects by reducing TLR4-mediated signaling and atrophy. Additionally, the study reveals a novel finding, by demonstrating that stimulated myotube contractions decrease membrane-bound TLR4 and increase the secretion of sTLR4 by myotubes.

Evaluation of cinnamaldehyde derivatives as potential protective agents against oxidative-stress induced myotube atrophy using chemical, biological and computational analysis

Skeletal muscle atrophy, associated with increased morbidity, mortality and poor quality of life, is a metabolic disorder with no FDA approved drug. Oxidative stress is one of the key mediators of atrophy that influences various cell signaling molecules. The goal of this study is to identify potential antioxidant agents that could be used to treat atrophy. In this study in vitro and in situ screening of different cinnamaldehyde (CNA) derivatives for their antioxidant effects was done along with computational analysis to understand the relationship between their chemical structure and biological activity.Data show that 2-hydroxycinnamaldehyde (2HCNA) worked better than other CNA analogues at physiological pH, while 4-Fluoro-2-methoxycinnamaldehyde (4FoCNA) showed the maximum antioxidant activity under acidic conditions. However, these derivatives (2HCNA and 4FoCNA) were found to be toxic to the cultured myotubes (mature myofiber) under both physiological and pathophysiological conditions. Immunofluorescence, bright-field microscopic and biochemical studies conducted using live C2C12 cells showed that pre-incubation with other CNA analogues i.e. 2-methoxycinnamaldehyde (2MeCNA) and 2-benzyloxycinnamaldehyde (2BzCNA) not only maintained the normal morphology of myotubes but also protected them from H2O2-induced atrophy. These compounds (2MeCNA and 2BzCNA) showed higher stability and antioxidant potential, as indicated by computer simulation data analyzed by Density Functional Theory (DFT) based molecular modeling. Overall, the chemical, biological, and computational studies reveal the therapeutic potential of CNA analogues (BzCNA and MeCNA) against oxidative-stress induced muscle atrophy in C2C12 cells.

Autoimmune Gastritis Accompanied by a Schwannoma Presenting as a Subepithelial Tumor

Autoimmune gastritis (AIG), a chronic inflammatory disease occurs as a result of a complex interaction between host-related and environmental factors. AIG may progress to severe atrophic gastritis secondary autoimmune-mediated parietal cell destruction in the stomach. AIG can be diagnosed based on anti-parietal cell antibody tests and endoscopy, which reveals widespread gastric corpus atrophy in patients with low serum pepsinogen I levels, a low pepsinogen I/II ratio, and elevated serum gastrin levels on serological testing. Tissue biopsy findings, which include mucosal atrophy and lymphocytic infiltration of the lamina propria may be useful for diagnostic confirmation. Decreased gastric acid secretion causes hypergastrinemia and enterochromaffin-like (ECL) cell proliferation, which can lead to neuroendocrine tumor development. Additionally, an autoimmune response results in parietal and chief cell injury, and proliferating ECL cells are detected in the deep mucosal layers in patients with AIG. Therefore, this condition may easily be misdiagnosed as a subepithelial tumor, and establishing a differential diagnosis for other types of subepithelial tumor accompanied by AIG is challenging. We present the case of a 54-year-old woman who was diagnosed with AIG with a concomitant subepithelial tumor based on serologic tests and biopsy findings and underwent wedge resection, which confirmed diagnosis of a schwannoma.

#4277 EGF, GDF-15 AND NEOPTERIN AS PREDICTORS OF CHRONIC KIDNEY DISEASE PROGRESSION IN CHILDREN BY MEANS OF ARTIFICIAL NEURAL NETWORK MODELS

Abstract Background and Aims Progression of chronic kidney disease (CKD) is a compound process, where activation of immunocompetent cells and subclinical inflammation play pivotal role. Enhanced atrophy of the tubular cells, and finally, gradual fibrosis of tubulointerstitial tissue, are responsible for irreversible character of the disease. Multiple molecules influence above-mentioned processes. Growth differentiation factor (GDF)15, a member of TGF-β cytokine superfamily, is a marker of inflammation and an integrative signal in both acute and chronic stress conditions. Elevated serum concentrations of GDF15 were associated with increased risk of development and progression of CKD in adults, as well as with mortality in this group of patients. Our previous investigation revealed increased serum GDF15 concentrations in children on chronic dialysis. Epidermal growth factor (EGF), a tubule-specific protein, promotes proliferation, differentiation and migration of epithelial cells, and therefore, modulates regeneration of injured renal tubules. Decreased concentrations of EGF in urine were observed in variety of kidney diseases, including diabetic nephropathy, lupus nephritis or CKD. Our previous analysis of EGF serum concentrations in CKD children confirmed their decreased values on chronic dialysis. Neopterin is a product of activated monocytes and macrophages and serves as a marker of cell-mediated immunity. Elevated serum concentrations of neopterin were observed in CKD adult patients, our investigation revealed its increased values in children on chronic dialysis. None of the above mentioned markers was analyzed in the population of CKD children treated conservatively. Therefore, the aim of study was to assess the serum concentrations of EGF, GDF-15 and neopterin in children with CKD on conservative treatment and verify the usefulness of these markers in predicting CKD progression by means of artificial intelligence tools. Method The study group consisted of 153 children with pre-dialysis CKD stages 1-5 (stage 1 – 27 patients, stage 2 – 26 patients, stage 3 – 51 patients, stage 4 - 28 patients, stage 5 – 21 patients). EGF, GDF-15 and neopterin serum concentrations were assessed by ELISA. The patient database was implemented into the artificial neural network. In detail, the recursively selected subsets of input variables constituted the input layer of an artificial neural network built of perceptrons (multi-layer perceptron). Anthropometric data, biochemical parameters, EGF, GDF15 and neopterin were included into the model, serum creatinine and eGFR, as direct classifiers of CKD stage, were excluded. Various models were tested, regarding their accuracy, AUROC and Matthews correlation coefficient (MCC) values. Results EGF serum concentrations decreased gradually, whereas GDF15 and neopterin values rose systematically with CKD progression, keeping statistically significant inter-stage differences. Moreover, the most precise ANN model, among the tested artificial neural networks, contained EGF, GDF15 and neopterin as input parameters and classified patients into either CKD 1-3 or CKD 4-5 groups. This model has put new patients into appropriate classes with excellent Accuracy of 96.77%, AUROC 0.9169 and Matthews correlation coefficient (MCC) of 0.9157. Conclusion The presented model of an artificial neural network, with serum concentrations of EGF, GDF15 and neopterin as input parameters, may serve as a useful predictor of CKD progression in the pediatric population. It suggests the essential role of inflammatory processes, defined by newly discovered markers, in the renal function decline towards advanced stages of CKD in children.

The effect of smoking and electronic cigarettes on rat testicles

ObjectiveAfter the negative effects of smoking on public health were proven, smoking cessation campaigns were initiated by health ministries and non-governmental organizations. Many drugs have been tried to reduce the addiction to smoking and the nicotine contained in it. Recently, e-cigarettes (EC) are widely used for smoking cessation efforts, although the effects and possible harms are not fully known. In our study, we planned to show the effect of cigarette and EC smoke on the male urogenital system. MethodsAdult male wistar rats were exposed to cigarette and EC smoke in a specially designed glass bell jar. Urine cotinine levels, testicular weights, gonadosomatic index, sperm count and sperm motility, testicular histology, and biochemical findings were compared with the control group. ResultsIn some rats in the cigarette and EC group, the seminiferous tubules were disorganized, and the germ cells and Sertoli cells were separated and shed. Stopped germ cell separation, cavity formation, necrosis, fibrosis, and atrophy were observed in severe cases. Higher PCO levels were found in the cigarette group compared to controls. Tissue homogenates levels of LPO were higher in both EC and cigarette groups compared to controls. No significant differences were observed between groups in terms of sperm motility and sperm count. ConclusionCigarette and EC liquid can increase oxidative stress as well as cause morphological changes in the testicle. To be a safe option in smoking cessation studies, its effect on people needs to be enlightened.

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

ObjectiveThe present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats. MethodsTwenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment. ResultsIntragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC. ConclusionThe present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.

Anthocyanins from Malus spp. inhibit the activity of Gymnosporangium yamadae by downregulating the expression of WSC, RLM1, and PMA1.

Malus plants are frequently devastated by the apple rust caused by Gymnosporangium yamadae Miyabe. When rust occurs, most Malus spp. and cultivars produce yellow spots, which are more severe, whereas a few cultivars accumulate anthocyanins around rust spots, forming red spots that inhibit the expansion of the affected area and might confer rust resistance. Inoculation experiments showed that Malus spp. with red spots had a significantly lower rust severity. Compared with M. micromalus, M. 'Profusion', with red spots, accumulated more anthocyanins. Anthocyanins exhibited concentration-dependent antifungal activity against G. yamadae by inhibiting teliospores germination. Morphological observations and the leakage of teliospores intracellular contents evidenced that anthocyanins destroyed cell integrity. Transcriptome data of anthocyanins-treated teliospores showed that differentially expressed genes were enriched in cell wall and membrane metabolism-related pathways. Obvious cell atrophy in periodical cells and aeciospores was observed at the rust spots of M. 'Profusion'. Moreover, WSC, RLM1, and PMA1 in the cell wall and membrane metabolic pathways were progressively downregulated with increasing anthocyanins content, both in the in vitro treatment and in Malus spp. Our results suggest that anthocyanins play an anti-rust role by downregulating the expression of WSC, RLM1, and PMA1 to destroy the cell integrity of G. yamadae.

Roflumilast alleviates adenine-induced chronic kidney disease by regulating inflammatory biomarkers

The present investigation was carried out to explore the role of roflumilast, a PDE4 inhibitor, as a potential treatment option for chronic kidney disease. Forty-six male Wistar rats were divided into five groups: Control, Disease control (50 mg/kg Adenine p.o.), Adenine + Roflumilast (0.5, 1 and 1.5 mg/kg, p.o.). Various urinary and serum biomarkers, antioxidant status, histopathology, and protein expression of inflammatory markers were measured to investigate the effects of roflumilast on kidney functions. Adenine was found to elevate the levels of serum creatinine, urea, uric acid, sodium, potassium, chloride, magnesium, and phosphorus and reduce the level of serum calcium. Further, adenine significantly increased the serum TGF-β levels and reduced the anti-oxidant indices. Significant elevation was observed in protein expression of IL-1β, TNF-α, MCP-1, ICAM-1, and Fibronectin. Histopathologically, adenine caused thickening of the glomerular basement membrane, inflammatory cells infiltration, atrophy, and glomeruli deterioration. However, Roflumilast administration (1 mg/kg) remarkably decrease serum creatinine, urea, uric acid, sodium, potassium, chloride, magnesium, phosphorus by 61%, 40%, 44%, 41%, 49%, 58%, 59% and 42% respectively, and increase in calcium by 158%. Moreover, Roflumilast (1 mg/kg) significantly reduced serum TGF-β levels by 50% and elevated anti-oxidant indices by 257%, 112%, and 60%, respectively. The protein expression was significantly reduced by 5.5-fold, 7-fold, 5.7-fold, 6.2-fold, and 5.1-fold individually. Roflumilast noticeably improved the structure of glomeruli, tubules, and cellular functioning. The study confirmed that Roflumilast has the potential to ameliorate renal injury by reducing and regulating inflammatory responses.

Acinar Cell Proliferation Promoted by BMP2 in Injured Mouse Parotid Gland: BMP2 Promotes Cell Proliferation in Parotid Gland.

To identify factors that affect salivary gland recovery, we investigated the expression and function of bone morphogenetic protein 2 (BMP2) in mice. Using a micro clip, mice parotid glands were removed 7 days after the ligation of the unilateral parotid excretory duct. Thereafter, they were weighed and stained with hematoxylin and eosin, and BMP2 expression was examined via real-time reverse transcription-polymerase chain reaction. Primary cultures of parotid glands were prepared, and BMP2 protein was added to the culture medium for 48 hr to examine its effect on cell proliferation. E-cadherin and vimentin expression was examined using western blotting. Finally, immunohistochemical staining using an anti-Ki67 antibody was performed. Duct-ligated parotid glands weighed less than those that were collected after sham surgery and showed acinar cell atrophy. They also showed higher BMP2 expression than control glands. Primary-cultured parotid acinar cells supplemented with BMP2 showed higher proliferative potential than control cells. Furthermore, they showed E-cadherin, but not vimentin, expression, and their percentage of Ki67-positive cells were higher than that corresponding to the controls. Injury to salivary glands by excretory duct ligation increased BMP2 expression, which may be involved in maintaining salivary gland function by inducing acinar cell proliferation.

Base editing rescue of spinal muscular atrophy in cells and in mice.

Spinal muscular atrophy (SMA), the leading genetic cause of infant mortality, arises from survival motor neuron (SMN) protein insufficiency resulting from SMN1 loss. Approved therapies circumvent endogenous SMN regulation and require repeated dosing or may wane. We describe genome editing of SMN2, an insufficient copy of SMN1 harboring a C6>T mutation, to permanently restore SMN protein levels and rescue SMA phenotypes. We used nucleases or base editors to modify five SMN2 regulatory regions. Base editing converted SMN2 T6>C, restoring SMN protein levels to wild type. Adeno-associated virus serotype 9-mediated base editor delivery in Δ7SMA mice yielded 87% average T6>C conversion, improved motor function, and extended average life span, which was enhanced by one-time base editor and nusinersen coadministration (111 versus 17 days untreated). These findings demonstrate the potential of a one-time base editing treatment for SMA.

5′-CMP and 5′-UMP alleviate dexamethasone-induced muscular atrophy in C2C12 myotubes

Atrogin-1 and muscle RING finger 1 (MuRF1) are ubiquitin ligases specifically expressed during skeletal muscle atrophy and mediate muscle protein degradation. In contrast, PGC-1α (peroxisome proliferator-activated receptor γ coactivator 1α), which is a master regulator of mitochondrial biosynthesis, protects skeletal muscle from atrophy. Pyrimidine nucleoside 5′-monophosphates, such as cytidine 5′-monophosphate (5′-CMP) and uridine 5′-monophosphate (5′-UMP), induce PGC-1α expression and promote myotube formation in mouse C2C12 cells. In this study, we determined the effect of 5′-CMP and 5′-UMP on muscular atrophy in C2C12 myotube cells. 5′-UMP decreased Atrogin-1 and MuRF1 mRNA levels that were upregulated by dexamethasone treatment. 5′-CMP and 5′-UMP ameliorated dexamethasone-mediated atrophy in C2C12 myotubes. Furthermore, the combination of 5′-CMP and 5′-UMP further alleviated dexamethasone-mediated atrophy. In addition, cytidine and uridine, the precursors of 5′-CMP and 5′-UMP, markedly ameliorated dexamethasone-mediated atrophy. Considering nucleotide metabolism and absorption, the active metabolites underlying the observed effects of 5′-CMP and 5′-UMP appear to be cytidine and uridine. Our results indicate that 5′-CMP alleviates muscle atrophy by activating PGC-1α and differentiation, and 5′-UMP alleviates muscle atrophy by suppressing the activation of the myolytic system, whereas the combined use of both enhances the muscle atrophy inhibitory effect. 5′-CMP and 5′-UMP may be an effective and safe treatment for muscular atrophy.

Effects of Single and Combined Exposure to Environmentally Relevant Concentrations of Cyfluthrin and Copper on Digestive Gland Histology of Lymnaea stagnalis

Pyrethroid insecticides and heavy metals frequently co-exist in aquatic systems, due to intensive anthropogenic activities, and their effects on aquatic organisms are needed to be investigated. In the present work, single and combined effects of environmentally realistic concentrations of cyfluthrin and copper on the digestive gland histology of the freshwater pond snail (Lymnaea stagnalis) were evaluated. Snails were exposed to 1 µg/L cyfluthrin, 10 µg/L copper, and 1 µg/L cyfluthrin + 10 µg/L copper mixture for 96 h. Cyfluthrin-exposed samples showed apparent vacuolization, basal lamina separations, and disrupted digestive cells. Exposure to copper alone gave rise to enlargements of the tubule lumens and the intertubular area, degenerated tubules, atrophied basophilic cells, prominently disrupted and ruptured digestive cells, and nuclear enlargements in some basophilic cells. Cyfluthrin and copper mixture caused more severe histopathological changes in the digestive gland. General tissue appearance was altered by prominently degenerated, and fused tubules lacking cellular structure; tubule lumens filled with cellular content were noted. Increased intertubular regions were filled with connective tissue formations. Progressive disruption of digestive cells; and basophilic cell atrophy were also observed. The findings of the current study highlighted that cyfluthrin and copper at environmentally relevant concentrations caused a biological response in L. stagnalis; however, this response was more intense following their co-exposure.

Testicular histopathology in rats co-exposed to heat and psychological stressors

ObjectivesThe present study aimed to investigate the effect of co-exposure to heat and psychological stressors on testicular tissue as one of the major male reproductive organs in rats. MethodsForty adult male Wistar rats were divided into four groups of 10 animals each. The first group was exposed to heat stress (Temperature Humidity Index: 57.75 ± 3), the second group was exposed to three psychological stressors including strobe light ultrasonic sound, and tilting cage, and the third group was co-exposed to both heat and psychological stress daily. The order of exposure to various psychological stressors was randomized. Following the last day of the 40 -day exposure, the rats were euthanized and their testicular tissues were fixed in Bouin's solution. Moreover, a tissue processor, a microtome as well as hematoxylin, and eosin staining were used for tissue preparation. ResultsThe co-exposure to psychological and heat stress can cause negative testis histopathological changes including spermatid retention, degeneration of round spermatids and spermatocytes, degeneration and depletion of elongated Spermatid, Sertoli cell status, tubular degeneration/atrophy, Leydig cell atrophy, tubular dilatation, re-tubular dilation, and sclerosis status in a number of rats. Moreover, the histopathological changes were higher in the group exposed to heat stress compared with those exposed to psychological stress. ConclusionsAlthough exposure to either stressor alone can affect testicular tissue as part of the reproductive system, co-exposure to both stressors may result in an increased risk of adverse effects on testicular tissue.

Xenogeneic transplantation of mitochondria induces muscle regeneration in an in vivo rat model of dexamethasone-induced atrophy.

Muscle atrophy significantly impairs health and quality of life; however, there is still no cure. Recently, the possibility of regeneration in muscle atrophic cells was suggested through mitochondrial transfer. Therefore, we attempted to prove the efficacy of mitochondrial transplantation in animal models. To this end, we prepared intact mitochondria from umbilical cord-derived mesenchymal stem cells maintaining their membrane potential. To examine the efficacy of mitochondrial transplantation on muscle regeneration, we measured muscle mass, cross-sectional area of muscle fiber, and changes in muscle-specific protein. In addition, changes in the signaling mechanisms related to muscle atrophy were evaluated. As a result, in mitochondrial transplantation, the muscle mass increased by 1.5-fold and the lactate concentration decreased by 2.5-fold at 1 week in dexamethasone-induced atrophic muscles. In addition, a 2.3-fold increase in the expression of desmin protein, a muscle regeneration marker, showed a significant recovery in MT 5µg group. Importantly, the muscle-specific ubiquitin E3-ligases MAFbx and MuRF-1 were significantly decreased through AMPK-mediated Akt-FoxO signaling pathway by mitochondrial transplantation compared with the saline group, reaching a level similar to that in the control. Based on these results, mitochondrial transplantation may have therapeutic applications in the treatment of atrophic muscle disorders.

The effect of neuronal atrophy on the pathogenesis of Huntington's disease.

Computational models play a pivotal role in the integration of complex biochemical reaction kinetics with the subcellular organization to obtain a deeper understanding of the cellular response to a drug. We have created a realistic 3D reconstruction of a segment of the axon including the endoplasmic reticulum and the subcellular arrangement of molecules involved in calcium homeostasis and calcium-induced calcium release. The dynamics of the system are modeled using MCell4, a stochastic, particle-based Monte Carlo simulator. We studied the effect of cell atrophy on the disease state of Huntington's disease (HD), a fatal neurological disorder that has no cure yet. HD is characterized by motor dysfunction, mental impairment, and cognitive decline. At the molecular level, HD is caused by a mutation in the first exon of the gene encoding the Huntingtin protein resulting in an expanded polyglutamine. Calcium dyshomeostasis is believed to be one of the key causes of HD. In particular, defective RyR function has been reported in HD leading to elevated intracellular Ca2+ levels. One of the drugs currently under development that can potentially be used for HD is a small-molecule drug RycalTM that may act by stabilizing leaky ryanodine receptor (RyR)/Ca2+ release channels that are in development for a severe form of muscle atrophy. Interestingly, our model indicates that even if all receptors and calcium pumps work normally, cell atrophy alone could lead to the “leaky RyR” receptors phenomenon, and will further result in saturation of the calcium buffer, calbindin, and accumulation of intercellular calcium which can be quite toxic to the cell. The computer model that we have built can be further developed to help us understand whether drugs in development or proposed for HD might be worthwhile or not in an effort to treat/cure HD.

Biological Assessment of Stevioside and Sucralose as Sucrose Substitutes for Diabetics on STZ-Induced Diabetes in Rats.

Numerous food organizations have identified excessive calorie consumption and accompanying ailments as significant health risks associated with high sugar consumption. Administering stevioside (ST), sucralose (SU), and the two synergically (SU+ST) affected normal rats' weight gain. In the current study, SU showed the highest undesired effect. Indeed, administering the three treatments to diabetic rats (DR) did not improve the rats' weight gain. Although, insulin injection synergically with the treatments improved the weight gain, as recorded after three weeks. The best-improving rate was observed in the ST group. After the administration of ST and ST+SU to the DR, the blood glucose level (GL) was positively affected, with SU having no effects on reducing the GL. A considerable reduction in serum insulin (SIL) was noted in the DR+SU group. On the contrary, ST did not negatively affect the SIL, rather an improvement was recorded. In addition, giving SU did not significantly affect the ALT level in the DR or normal rats (NR). A significant improvement in total bilirubin (TBILI) was observed when insulin was injected with ST or SU in DR groups. Further, triglycerides (TG) after administering ST, SU, or ST+SU to NR had no significant difference compared to the control group (NR). Although, the three treatments markedly but not significantly lowered TG in the DR. For total cholesterol (CHO), both DR and NR had no significant effect after the three treatments. No histopathological alterations were recorded in the NR group. Diffuse and severe atrophy of the islands of Langerhans due to depletion of their cells and mild papillary hyperplasia of the pancreatic ducts were represented by a slightly folded ductal basement membrane and newly formed ductules in STZ-DR. Simultaneous atrophy and absence of the cells of islands of Langerhans besides ductal hyperplasia were evident in DR+SU. Hyperplastic ductal epithelium and atrophic Langerhans cells were seen in DR+SU+In. Degeneration and mild atrophy were observed in the islands of Langerhans structures. There was essentially no noticeable change after utilizing ST. A slight shrinkage of the Langerhans' islets was detected in DR+ST. In DR+ST+In, no histopathological alterations in the islands of Langerhans were recorded. Congestion in the stromal blood vessels associated with degenerative and necrotic changes in the cells of the islands of Langerhans in DR+SU+ST was observed. In NR+SU, congestion of the blood vessels associated with mild atrophy in the islands of Langerhans and dilatation in stromal blood vessels was noticed. In conclusion, ST is safe, and SU should be taken cautiously, such as mixing with ST and/or taken at a very low concentration to avoid its drastic effect on the human body.

Gastrodin attenuates renal injury and collagen deposition via suppression of the TGF-β1/Smad2/3 signaling pathway based on network pharmacology analysis.

Background: Gastrodin has been widely used clinically in China as an antihypertensive drug. However, its effect on hypertensive renal injury is yet to be elucidated. The current study aimed to investigate the effects of gastrodin on hypertensive renal injury and its underlying mechanisms by network pharmacology analysis and validation in vivo and in vitro. Methods: A total of 10 spontaneously hypertensive rats (SHRs) were randomly categorized into the following two groups: SHR and SHR + Gastrodin groups. Wistar Kyoto (WKY) rats were used as the control group (n = 5). The SHR + Gastrodin group was intragastrically administered gastrodin (3.5mg/kg/day), and the rats in both WKY and SHR groups were intragastrically administered an equal amount of double-distilled water for 10 weeks. Hematoxylin-eosin, Masson's trichrome, and Sirius red staining were used to detect the pathological changes and collagen content in the renal tissues. Network pharmacology analysis was performed to explore its potential targets and related pathways. In vitro, the CCK-8 assay was used to determine the cell viability. Immunohistochemistry and western-blotting analyses were employed to assess the protein expression associated with renal fibrosis and transforming growth factor-β1 (TGF-β1) pathway-related proteins in the renal tissues or in TGF-β1-stimulated rat kidney fibroblast cell lines (NRK-49F). Results: Gastrodin treatment attenuates renal injury and pathological alterations in SHRs, including glomerular sclerosis and atrophy, epithelial cell atrophy, and tubular dilation. Gastrodin also reduced the accumulation of collagen in the renal tissues of SHRs, which were confirmed by downregulation of α-SMA, collagen I, collagen III protein expression. Network pharmacology analysis identified TGFB1 and SMAD2 as two of lead candidate targets of gastrodin on against hypertensive renal injury. Consistently, gastrodin treatment downregulated the increase of the protein expression of TGF-β1, and ratios of both p-Smad2/Smad2 and p-Samd3/Smad3 in renal tissues of SHRs. In vitro, gastrodin (25-100μM) treatment significantly reversed the upregulation of α-SMA, fibronectin, collagen I, as well as p-Smad2 and p-Smad3 protein expressions without affecting the cell viability of TGF-β1 stimulated NRK-49F cells. Conclusion: Gastrodin treatment significantly attenuates hypertensive renal injury and renal fibrosis and suppresses TGF-β1/Smad2/3 signaling in vivo and in vitro.