Histomorphological Changes Research Articles

Histological study of ovary of indian major carp Labeo rohita (ham), in pond condition

The scientific management for obtaining high yield of fish production eventually calls for the adequate and indepth study of breeding mechanism. Month wise variations with reference to histomorphological changes in oocytes of a fresh water fish, Labeo rohita (Ham.) have been described. The different stages of oocytes development were examined microscopically. Five stages of oocytes development of Labeo rohita under study were identified. The matured ova were found in the month of May, June and July in the ovary of this fish.. KEYWORDS :Histology, Ovary, Labeo rohita (Ham.)

Acupotomy ameliorates knee osteoarthritis-related collagen deposition and fibrosis in rabbit skeletal muscle through the TGF-β/Smad pathway

ObjectiveTo investigate the effects of acupotomy on skeletal muscle fibrosis and collagen deposition in a rabbit knee osteoarthritis (KOA) model. MethodsRabbits (n = 18) were randomly divided into control, KOA, and KOA + acupotomy (Apo) groups (n = 6). The rabbits in the KOA and Apo groups were modeled using the modified Videman's method for 6 weeks. After modeling, the Apo group was subjected to acupotomy once a week for 3 weeks on the vastus medialis, vastus lateralis, rectus femoris, biceps femoris, and anserine bursa tendons around the knee. The behavior of all animals was recorded, rectus femoris tissue was obtained, and histomorphological changes were observed using Masson staining and transmission electron microscopy. The expression of transforming growth factor-β1 (TGF-β1), Smad 3, Smad 7, fibrillar collagen types I (Col-I) and III (Col-III) was detected using Western blot and real-time polymerase chain reaction (RT-PCR). ResultsHistological analysis revealed that acupotomy improved the microstructure and reduced the collagen volume fraction of rectus femoris, compared with the KOA group (P = .034). Acupotomy inhibited abnormal collagen deposition by modulating the expression of fibrosis-related proteins and mRNA, thus preventing skeletal muscle fibrosis. Western blot and RT-PCR analysis revealed that in the Apo group, Col-I, and Col-III protein levels were significantly lower than those in the KOA group (both P < .01), same as Col-I and Col-III mRNA levels (P = .0031; P = .0046). Compared with the KOA group, the protein levels of TGF-β1 and Smad 3 were significantly reduced (both P < .01), as were the mRNA levels of TGF-β1 and Smad 3 (P = .0007; P = .0011). Conversely, the levels of protein and mRNA of Smad 7 were significantly higher than that in the KOA group (P < .01; P = .0271). ConclusionAcupotomy could alleviate skeletal muscle fibrosis and delay KOA progress by inhibiting collagen deposition through the TGF-β/Smad pathway in the skeletal muscle of KOA rabbits.

Effect of solid-state fermentation of Brewer's spent grain on digestibility and digestive function of european seabass (Dicentrarchus labrax) juveniles

This study assessed the effects of dietary inclusion of solid-state fermented (SSF) brewer’s spent grain (BSG) with Aspergillus ibericus on nutrient and energy digestibility, digestive enzyme activity, and intestinal histomorphology of juvenile European seabass (Dicentrarchus labrax). Five diets (18 % crude lipids and 45 % crude protein) were formulated, including a control diet (without BSG), two diets with 10 % and 20 % unfermented BSG (10BSG and 20BSG), and two other diets with 10 % and 20 % fermented BSG (10BSG-SSF and 20BSG-SSF). SSF affected the BSG's nutritional composition, including a 21 % increase in protein content and reductions in lipid (49 %), cellulose (30 %), hemicellulose (34 %), and lignin (7.3 %) content. Antioxidant activity and total phenolic compounds were minimal before SSF but significantly increased after SSF. Dietary incorporation of 20 % BSG-SSF increased the digestibility of dry matter (p<0.01), protein (p=0.03), isoleucine (p=0.03), glutamate (p=0.02), lipids (p=0.04) and energy (p=0.04) compared to the 20BSG diet. Moreover, SSF also modulated digestive enzyme activity, reducing total protease (p=0.03) and trypsin (p=0.01) activities in fish fed the 10BSG-SSF diet compared to the control diet. Fish fed the 20BSG diet showed changes in intestinal histomorphology compared to those fed the control diet, and SSF of BSG appeared to mitigate these effects.Overall, these results indicate that SSF is a promising technique for enhancing the nutritional quality of low-value agro-industrial by-products, such as BSG, increasing their potential use as feed ingredients and contributing to reducing the climate and environmental footprint of aquaculture production.

Histomorphological assessment of the germanium-doped calcium phosphate ceramics on reparative osteogenesis in rabbits with systemic osteoporosis

Bone fractures are mostly urgent in nature, complex in pathogenetic, diagnostic and therapeutic aspects, and reparative osteogenesis is multimodal and depends on the balanced and reciprocal interaction of many factors. The results of histological studies for osteoreplacement of bone defects in rabbits with systemic osteoporosis are presented. The purpose of the work is histomorphological evaluation of bone regenerates after osteoreplacement with germanium-doped hydroxyapatite ceramics in rabbits with secondary osteoporosis. Experimental osteoporosis in rabbits (n=18) was induced by administration of 0.4% dexamethasone solution. In animals of the experimental group, bone defects were replaced with granules of hydroxyapatite ceramics doped with germanium, and in animals of the control group, they healed under a blood clot. Histological sections were made on a rotary microtome with a thickness of 5 to 10 μm and stained with Weigert's iron hematoxylin and 1% alcohol-based eosin solution (manufactured by Diapath, Italy). On the 60th day of reparative osteogenesis in the animals of the experimental group, the site of the bone defect was flled with compact bone tissue with minor remnants of spongy bone tissue. Slightly expanded Haversian canals were noted. In the control group, the defect site was flled with coarse and spongy bone tissue. Bone beams of various thicknesses with a small number of osteoblasts and single walled osteocytes were visualized. A signifcant number of osteocyte lacunae were empty due to the process of osteocyte lysis. Haversian canals are signifcantly dilated with a small number of vessels. Also, lacunae of bone tissue resorption and acellular areas were observed on histological specimens. Histomorphological assessment of bone regenerates confrms the realization of osteoconductive, osteointegrative and osteoinductive properties of calcium-phosphate ceramics doped with germanium in conditions of osteoporotic fractures of tubular bones. Keywords: systemic osteoporosis, bone fractures, cancellous and compact bone tissue, histological sections, histomorphological changes, rabbits.

Exposure to atrazine and endosulfan alters oviductal adenogenesis in the broad-snouted caiman (Caiman latirostris).

The molecular pathways involved in oviductal adenogenesis are highly conserved among vertebrates. In this work, we study the histomorphological changes and molecular pathways involved in Caiman latirostris oviductal adenogenesis and the effects of in ovo exposure to environmentally relevant doses of endosulfan (END) and atrazine (ATZ) on these processes. To this end, the histomorphological changes at epithelial and subepithelial compartments, the protein expressions of β-catenin and Wnt-7a, and the gene expression of metalloproteinases (MMPs) and its inhibitors (TIMPs) were evaluated as biomarkers of oviductal adenogenesis in prepubertal juvenile C. latirostris. Exposure to END altered adenogenesis-related epithelium characteristics and mRNA expression of MMP2, MMP9, and TIMP1. Exposure to ATZ increased the width of the subepithelial stroma with loosely arranged collagen fibers and increased β-catenin expression in buds (invaginated structures that precede glands). The results demonstrate that in ovo exposure to ATZ and END alters oviductal adenogenesis at tissue, cellular, and molecular levels. An altered oviductal adenogenesis could impair fertility, raising concern on the effects of pesticide pollution in wildlife and domestic animals.

Betaine Attenuates The Expression of Vasoactive Mediators and Histological Alterations Associated with Ovarian Hyperstimulation Syndrome in Rats.

Ovarian hyperstimulation syndrome (OHSS) is one female reproductive disorder that can occur after administration of injectable hormonal drugs to stimulate ovulation. Betaine (BET) is an intracellular biomolecule with anti-inflammatory and tissue protective effects. There is no information about its effects in an experimental model of OHSS. The current study aims to investigate the possible effects of BET on abnormal expressions of vasoconstrictor proteins and ovarian histological changes in an experimental OHSS rat model. In this experimental study, 30 adult female rats (two months old) were randomly divided into six groups (n=5 per group): i. Control, ii. OHSS [10 IU sc equine chorionic gonadotropin (eCG) for 4 days followed by 30 IU sc human chorionic gonadotropin (hCG) on the fifth day], iii. OHSS+BET (200 mg/kg/day, orally for seven days), iv. OHSS+Cabergoline (CAB, 100 mg/kg/day, orally for six days), v. BET, and vi. CAB. Expression levels of vascular endothelial growth factor (VEGF), cyclooxygenase-2 (COX-2), and blood levels of oestradiol (E2) and progesterone (P4) were measured at the end of the experiment. The ovaries were studied for histomorphological changes. Induction of OHSS altered tissue histology, including an increase in the number of corpora lutea and atretic follicles, and decreased the number of follicular reserves. In this group, we observed increased expressions of the VEGF and COX-2 proteins, and increased serum E2 and P4 levels. Administration of CAB and BET significantly attenuated all molecular and histological alterations observed in the OHSS animals. Our findings, for first time, indicate the beneficial effects of BET to reduce OHSS complications in patients by reducing the expressions of vasoactive proteins and improving changes to the ovarian tissues. The findings are similar to CAB and can be a new avenue for future research on BET.

FINERENONE TREATMENT PREVENTS GLOMERULAR DAMAGE AND RENAL INTERSTITIAL INFLAMMATION IN TYPE 1 DIABETIC RATS WITH CHRONIC KIDNEY DISEASE

Objective: Finerenone (FIN), a non-steroidal mineralocorticoid receptor antagonist, has shown a protective effect on kidney and cardiovascular damage progression in type 1 diabetic Munich Wistar Frömter rats with established chronic kidney disease (CKD). The gradual decline in kidney function in diabetic CKD is due to a thickening of glomerular and Bowman capsule basement membranes, mesangial matrix expansion, and tubulointerstitial fibrosis. In this study we aimed to characterize whether the renal protection of FIN is related to a decrease in kidney damage through histomorphological changes in diabetic MWF rats. Design and method: Diabetes was induced by a streptozotocin injection (15 mg/Kg, i.p.) together with exposure to a high fat/high sucrose (HF/HS) diet (MWF-D) for 6 weeks. The MWF-D-FIN group was additionally treated with 10 mg/Kg/day of FIN included in the HF/HS diet (n = 6/group). Morphometrical analysis of renal structural changes was performed under hematoxylin and eosin staining, and light microscopy. Matrix metalloproteinases (MMP)-2 and MMP-9 activities were determined by zymography. Results: MWF-D and MWF-D-EP rats exhibited significantly elevated glycemia, water intake, urine volume, glucosuria, and kidney weight compared to control MWF. Histological examination of kidney samples of MWF and MWF-D rats showed hypertrophic glomeruli with an increase of the glomerular tuft and the Bowman's space. These pathological alterations were prevented by FIN treatment. The percentage of glomerulosclerosis was significantly higher in MWF and MWF-D rats and 59% lower in the MWF-D-FIN group (p&lt;0.001) whereas the glomerulosclerosis index was reduced to 33% by FIN (p&lt;0.001). Moreover, MWF-D rats showed an increased interstitial inflammation index compared to MWF rats (p&lt;0.001) which was reduced to 44% (p&lt;0.001) in the MWF-D-FIN group. Since an increase in MMP-2 and MMP-9 activities are associated with fibrosis development, these were determined by zymography in kidney samples of all groups. Both MMP-2 and MMP-9 were significantly higher in MWF-D compared to MWF and decreased by FIN treatment. Conclusions: In conclusion FIN prevents the development of diabetic kidney damage due to a reduction in glomerular alterations and interstitial inflammation associated to an inhibition of MMP-2 and MMP-9 activities.

Histomorphological changes in gills and hepatopancreas of crabs: an integrative review

Crustaceans are a biological model of ecotoxicology and consequently are widely used to assess the impact of xenobiotics. The toxicity of xenobiotics starts in the interaction of biomolecules and can even cause changes in an entire ecosystem. This paper aims to compile the histomorphological changes caused by xenobiotics in the gills and hepatopancreas of crabs through an integrative review. The literature search was done using PubMed with keywords in English and the filters "full text" and "publications from 2001-21". A total of 299 studies were found and 35 of them were selected based on the title and abstract. Later, 17 were included in the study. There were 18 categories of histomorphological changes in the gills and 22 in the hepatopancreas. We noticed that there is no consensus on the terminology used for histomorphological changes in this group. Furthermore, it was observed that the quality of the histological slides was quite diverse, making the identification and comparison of changes in the images problematic. The aforementioned considerations justify the need for this study to address a gap and encourage future research to develop a guide of histomorphological changes in crabs and its use as a tool for environmental biomonitoring.

Blood flow restriction training does not negatively alter the mechanical strength or histomorphology of uninjured equine superficial digital flexor tendons.

Low load exercise training with blood flow restriction (BFR) has become increasingly used by human physical therapists to prescribe controlled exercise following orthopaedic injury; its effects on the equine superficial digital flexor tendon (SDFT), however, are unknown. To investigate outcomes of pressure specific BFR walking exercise on uninjured equine SDFT biomechanics and histomorphology. Controlled in vivo experiment. Four forelimbs of four horses were exposed to 40 BFR-walk sessions (10-min interval walking) on a treadmill over a 56-day study period with their contralateral forelimbs serving as untreated controls. Similarly, four forelimbs of four control horses were exposed to 40 sham cuff walk sessions. On study Day 56, all horses (n = 8) were humanely euthanised and forelimb SDFTs underwent non-destructive biomechanical testing and corresponding histomorphological analysis. Significance in biomechanical parameters between treatment groups was analysed using a mixed-effects ANOVA with Tukey's post-hoc tests. Statistically significant differences in SDFT stiffness for both first (p = 0.02) and last cycles (p = 0.03) were appreciated within the BFR treated group only, with BFR exposed forelimbs being significantly stiffer than the contralateral unexposed forelimbs. When normalised to cross-sectional area, no significant differences were appreciated among treatment groups in elastic modulus for the first (p = 0.5) or last cycles (p = 0.4). No histological differences were appreciated among treatment groups according to Bonar, Movin, or musculotendinous junction evaluation criteria. Short-term comparisons were performed in a small sample population without correlation to performance outcome measures. Optimal occlusion percentages and walk protocols remain unknown. This study demonstrated no negative impact of BFR on mechanical strength of the equine SDFT; however, evidence suggests that BFR results in increased tendon stiffness based on biomechanical testing and subsequent calculations. No consistent detrimental histomorphological changes were seen.

Experimental study on the inhibition of the NLRP3 signaling pathway with Shengsan Jiedu Huayu decoction to alleviate inflammatory injury in rats with acute-on-chronic liver failure

Objective: To observe the therapeutic effect of Shengsan Jiedu Huayu decoction in alleviating inflammatory liver injury in rats with acute-on-chronic liver failure (ACLF) and its effect on the activation intensity for the NLRP3 signaling pathway. Methods: 63 SD rats were randomly divided into a blank group, a model group, and low-, medium-, and high-dose groups of Shengsan Jiedu Huayu decoction (7.29 g/kg/d, 14.58 g/kg/d, and 29.16 g/kg/d). The ACLF rat model was replicated using carbon tetrachloride combined with d-galactosamine and lipopolysaccharide. Different dose gradients of the Shengsan Jiedu Huayu decoction were used for a five-day intervention treatment, and then rat serum and tissue samples were collected. A biochemical analyzer was used to detect the serum levels of ALT, AST, and TBIL in rats. ELISA was used to detect serum IL-18 and IL-1β content. HE staining was used to observe histomorphological changes in liver tissue. Immunohistochemistry was used to detect GSDMD expression in liver tissue. Western blot and PCR were used to detect NLRP3, Caspase1, ASC, TLR4, IL-1β, IL-18 protein, and mRNA expression levels.The groups were compared using analysis of variance and the rank-sum test. Results: Compared with the blank group, the model group's rat liver tissue was severely injured. Serum levels of ALT, AST, and TBIL, inflammatory factors IL-1β and IL-18, and the GSDMD protein expression level, NLRP3 expression level, TLR4, caspase 1, ASC, IL-1β, IL-18 protein, and mRNA (P<0.01) were all significantly increased in the model than the blank group (P<0.01). Additionally, compared with the model group, the low-, medium-, and high-dose groups of Shengsan Jiedu Huayu decoction had improved liver tissue injury in ACLF rats, while the serum levels of ALT, AST, TBIL, IL-1β, IL-18, liver tissue GSDMD protein, NLRP3, TLR4, caspase 1, and ASC expressions were all lower in the different dose gradients of the Shengsan Jiedu Huayu decoction than the model group, with the most evident reduction in the high-dose group (P<0.01). Conclusion: Shengsan Jiedu Huayu decoction can weaken the activation intensity of the NLRP3 signaling pathway, alleviate liver tissue pathological injury, reduce inflammatory factor release, and alleviate inflammatory liver injury in ACLF rats.

Hepatoprotective effect of jojoba oil against lead-induced toxicity in rats

Background Previous investigations showed that jojoba oil exhibited a protective effect against hepatotoxicity caused by different toxicants, however, to the best of our knowledge, no prior research has been done to determine the effectiveness of jojoba oil in protecting against lead toxicity. Objective This study assessed the hepatoprotective properties of jojoba oil against lead toxicity in rats. Materials and methods The study included four groups, each consisting of six Sprague Dawley male rats, and orally administered jojoba oil (JO group), lead acetate (LA group), and lead acetate plus jojoba oil (protective group) Results and conclusion The results showed that lead acetate-induced hepatotoxic effects were revealed by increased serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase activities with hepatic histomorphological changes. There was a significant increase in serum total cholesterol, triglycerides, and low-density lipoprotein-cholesterol, while levels of high-density lipoprotein-cholesterol significantly declined compared to normal rats. Additionally, lead acetate triggered oxidative damage of hepatocytes, evidenced by a significant increase of malondialdehyde levels and a decrease of reduced glutathione levels and activities of superoxide dismutase and glutathione S-transferase. Administration of lead was associated with a change in the distribution of cells over different cell cycle phases, characterized by a marked increase in the sub G1 cell population and a significant decrease in the G0/G1 cell population. Supplementation of jojoba oil with lead acetate relieved the toxic impacts of lead acetate with an enhancement of the liver enzyme activities, antioxidant status, lipid profile parameters, and histopathological alterations. In conclusion, jojoba oil might be an effective natural product that offers a promising preventive action towards lead-induced liver damage in rats.

Dental Rehabilitation of Patients with Oncological Diseases of the Maxillofacial Region Using Dental Mini-Implants

The traditional method of dental rehabilitation of Malignant diseases of the oral mucosa and maxillofacial region is the use of a removable plate prosthesis. The disadvantage of this prothesis is a significant volume, insufficient fixation and mobility, which leads to problems with speech function and taste sensitivity. As an alternative, rehabilitation of patients by prosthetics based on dental mini-implants was proposed, which allows implantation in the alveolar process of patients with adverse histomorphological changes in the alveolar bone after radiation exposure. The aim of the study is to analyze the effectiveness of dental rehabilitation of patients with acute respiratory infections using dental mini-implants. According to the results of the study, the average values of periotestometry ranged from –0.3 to 0.6 cu (p &lt; 0,002), the average level of interleukin-6 (IL-6) from 0.18 ± 0.23 to 3.8 ± 1.1 pg/ml, while in the postoperative period – from 4.45 ± 0.22 to 5.5 ± 1.1 pg/ml. (p &lt; 0,002), the average value of quality of life (OHIP-14) is 14 ± 1.2 points (p &lt; 0,004). The use of mini-implantation is the optimal method of restoring the function of chewing and aesthetics in patients who have suffered from cancer, within the permissible radiation dose (no more than 70 gy).

Beating the heart failure odds: long-term survival after myocardial ischemia in juvenile rainbow trout.

Salmonid fish include some of the most valued cultured fish species worldwide. Unlike most other fish, the hearts of salmonids, including Atlantic salmon and rainbow trout, have a well-developed coronary circulation. Consequently, their hearts' reliance on oxygenation through coronary arteries leaves them prone to coronary lesions, believed to precipitate myocardial ischemia. Here, we mimicked such coronary lesions by subjecting groups of juvenile rainbow trout to coronary ligation, assessing histomorphological myocardial changes associated with ischemia and scarring in the context of cardiac arrhythmias using electrocardiography (ECG). Notable ECG changes resembling myocardial ischemia-like ECG in humans, such as atrioventricular blocks and abnormal ventricular depolarization (prolonged and fragmented QRS complex), as well as repolarization (long QT interval) patterns, were observed during the acute phase of myocardial ischemia. A remarkable 100% survival rate was observed among juvenile trout subjected to coronary ligation after 24 wk. Recovery from coronary ligation occurred through adaptive ventricular remodeling, coupled with a fast cardiac revascularization response. These findings carry significant implications for understanding the mechanisms governing cardiac health in salmonid fish, a family particularly susceptible to cardiac diseases. Furthermore, our results provide valuable insights into comparative studies on the evolution, pathophysiology, and ontogeny of vertebrate cardiac repair and restoration.NEW & NOTEWORTHY Juvenile rainbow trout exhibit a remarkable capacity to recover from cardiac injury caused by myocardial ischemia. Recovery from cardiac damage occurs through adaptive ventricular remodeling, coupled with a rapid cardiac revascularization response. These findings carry significant implications for understanding the mechanisms governing cardiac health within salmonid fishes, which are particularly susceptible to cardiac diseases.

Pathomorphological changes in laboratory animals exposed to lethal doses of disinfec-tants

The use of disinfectants is a crucial aspect of preventive and health improvement measures for infectious diseases in farm and domestic animals. Regulatory documents require the determination of toxicity to macroorganisms, including the establishment of lethal doses and toxicity groups, during the development and registration of antimicrobial agents. This study aimed to investigate the histological changes in the internal organs of laboratory animals when determining lethal doses of innovative disinfectants. The experiments used domestic disinfectants containing glutaraldehyde as the active ingredient. Histological studies were conducted on the internal organs (kidney, liver, stomach, intestines, and spleen) of 75 laboratory animals using an Axioskop 40/40FL microscope (Carl Zeiss, Germany) with video microscopic photography. The methods used were under current standards. Hemodynamic disorders were observed in the renal tissue under the influence of lethal doses of aldehyde disinfectants. These disorders were characterized by capillary dilation and blood filling. Glomerular capillary dilation and overflow with blood cells were also detected. Additionally, stasis was observed in the lumen of the microcirculatory vessels throughout the entire length. The examination of histological sections from animal liver samples revealed a significant expansion of Dissé spaces, variations in the size of hepatocyte nuclei, beam decomposition and fragmentation, small acute perivascular hemorrhages, leukostasis in sinusoids, and hemodynamic disorders. The structure of the organ's beams was also disturbed, and a significant number of venous vessels were dilated and excessively filled with blood cells. Minor changes were detected in the stomach, including desquamation of the epithelial cells of the glands and their exfoliation into the gastric lumen, as well as circulatory disorders. Epithelial desquamation, blood vessel dilation, and signs of connective tissue edema were observed in the intestine. The kidneys exhibited signs of acute venous hemorrhage and stasis in vessels of various calibers, with the development of small acute parenchymal hemorrhages and localized lymphoid cell death in the white pulp. The prospect of further research is to investigate the histomorphological changes in the internal organs of laboratory animals when exposed to modern complex disinfectants with different active ingredients.

Protective effect of FKBP12 on dextran sulfate sodium-induced ulcerative colitis in mice as a tacrolimus receptor

Ulcerative colitis (UC) is a multifactorial intestinal disease with a high incidence. In recent years, there has been an urgent need for pleiotropic drugs with a clear biosafety profile. Tacrolimus (TAC) is an immunosuppressant with stronger in vivo effects and better gastrointestinal absorption and is considered a potential treatment for UC. FKBP12 is a mediator of TAC immunosuppression; however, it is unclear whether it can participate in the development of UC in combination with TAC. The purpose of this study is to preliminarily validate the function of FKBP12 by establishing dextran sulfate sodium (DSS)-induced UC model and TAC treatment. The results revealed that TAC was effective in alleviating DSS-induced UC symptoms such as body weight and disease activity index (DAI). TAC significantly protects colonic tissue and attenuates DSS-induced histomorphological changes. In addition, FKBP12 is down-regulated in the intestinal tissue of DSS-induced UC mice and in serum samples of UC patients. In conclusion, our study revealed that FKBP12 may act as a TAC receptor to have anti-inflammatory and protective effects on DSS-induced UC in mice, which will provide a new option for the treatment of UC.

Platelet-rich plasma-derived extracellular vesicles inhibit NF-κB/NLRP3 pathway-mediated pyroptosis in intervertebral disc degeneration via the MALAT1/microRNA-217/SIRT1 axis

BackgroundIntervertebral disc degeneration (IDD) is a main contributor to lower back pain, and compression stress-induced apoptosis of nucleus pulposus (NP) cells and extracellular matrix (ECM) degradation has been implicated in the IDD progression. The functions of platelet-rich plasma (PRP)-derived extracellular vesicles (PRP-EVs) in regulating these biological processes remain unclear in IDD. Here, we aimed to investigate the key role of long noncoding RNA (lncRNA) MALAT1 incorporated in PRP-EVs in IDD. MethodsTert-butyl hydroperoxide (TBHP)-induced damage in NP cells was treated with PRP-EVs extracted from healthy volunteers, followed by MTT, EdU, TUNEL, and Western blot assays. IDD mice were also treated with PRP-EVs. Histomorphological and pathological changes were evaluated. The pyroptosis of cells and the degradation of ECM were detected by ELISA and immunohistochemistry. We screened the differentially expressed lncRNAs in NP cells after PRP-EVs treatment by microarray analysis. The downstream targets of MALAT1 in NP cells were predicted and validated by rescue experiments. FindingsTBHP induction reduced cell proliferation and exacerbated pyroptosis and ECM degradation, and PRP-EVs inhibited TBHP-induced cell damage. PRP-EVs-treated mice with IDD had reduced Thompson scores, increased NP tissue content, and restored ECM. PRP-EVs upregulated MALAT1 expression in vivo and in vitro, whereas MALAT1 downregulation exacerbated NP cell pyroptosis and ECM degradation. MALAT1 upregulated SIRT1 expression by downregulating microRNA (miR)-217 in NP cells. SIRT1 blocked the NF-κB/NLRP3 pathway-mediated pyroptosis, thereby alleviating IDD. InterpretationPRP-EVs deliver MALAT1 to regulate miR-217/SIRT1, thereby controlling NP cell pyroptosis in IDD.

The changes in tissue histomorphology and quality of DNA in healthy human autopsied tissues stored at −20 °C and −150 °C

The aim of this study was to compare the changes in tissue histomorphology and DNA quality in six different healthy tissues (brain, heart, lung, liver, spleen and kidney) exempted during autopsy of healthy individuals and storage at −20 °C and −150 °C three month. Tissue samples were obtained, divided by tissue and temperature group, and for each sample, tissue histomorphology and DNA (isolated from all tissues in duplicated − 72 samples of DNA) quality were analysed. Morphology of tissue samples was studied using H&E staining. DNA was isolated using the phenol–chloroform-isoamyl alcohol method. To assess the concentration and purity of the DNA samples, we used a spectrophotometer to measure absorbance at wavelengths of 280 nm and 260 nm. The fragments of human telomerase reverse transcriptase (hTERT) gene were amplified from the DNA using PCR reaction and then visualised using the 2 % agarose gel. Samples stored at −150 °C sustained the highest degree of histomorphological damage, while samples stored at −20 °C were less degraded, compared to control. The liver samples stored at −20 °C had a mean DNA concentration (1030.4 ± 51.5 ng/μl) higher than the samples of liver tissue stored at −150 °C (497.4 ± 167.1 ng/μl) (p < 0.001). Other tissues did not have statistically significantly different DNA concentration at both temperatures. Liver samples at −20 °C had degraded DNA, showed as the absence of hTERT gene in most of samples. Other tissue samples in both temperature groups had unfragmented DNA. Storing tissue samples at −20 °C is not inferior in terms of DNA yield and integrity, and possibly superior for tissue histomorphology, comparing with samples stored at −150⁰C.

Fish meal replacement by Brachytrupes portentosusas for Oreochromis niloticus: Effect on growth, feed utilization, fatty acid profiles, hematology, and histological changes

In the aquaculture section, the scarcity of sustainable feed components is the major challenge. Insects may become an important, sustainable resource for expanding the raw material repertoire. Herein, this study evaluated the effects of fish meal (FM) replacement with giant cricket meal (GCM) on growth, feed utilization, fatty acids profiles, hematology, and histological changes of Nile tilapia. Five isonitrogenous and isolipidic diets were formulated with a FM-based diet (without GCM or GCM0), the other four diets were prepared to replace FM with 55, 111, 164, and 217 g/kg of GCM (referred to as GCM55 - GCM217). A total of 300 fish (approximately 7.27 ± 0.01 g/fish) were randomly allocated to triplicate groups with 20 fish per tank and fed until satiation twice daily. The results showed a linear and quadratic relationship were observed between growth parameters and dietary replacement of GCM levels. Although the total replacement of FM by GCM217 significantly diminished, but GCM55 – GCM164 replacement was higher than a control group and improved the feed conversion ratio. The condition factor was similar among dietary treatments, but the hepatosomatic index and viscerosomatic index were affected by GCM inclusion levels (P < 0.05). Whole-body dry matter and ash were linearly and quadratically increased up to GCM164, and slightly decreased in the GCM217 group. Dietary GCM inclusion regulated whole-body composition in a dose-dependent manner. Muscle fatty acids were affected by dietary GCM replacement with a linearly decreased in 18:3n-3, 20:5n-3, and 22:6n-3, and the quadratic effect was observed for 22:6n-3. Dietary inclusion of GCM quadratically increased hemoglobin, total protein, albumin, and globulin, and linearly improved red blood cell and hemoglobin levels. However, GCM inclusions higher than 164 g/kg were linearly and quadratically increased aspartate aminotransferase and alanine aminotransferase compared to a control group. A mild degree of vacuole enlargement was found in GCM164, while a high degree of irregular shape, cytoplasm vacuolization, and swelling was found in GCM217 group. In addition, intestinal morphometrics of GCM217 group detected a significantly linear and quadratic decrease in villi height. Results of the present study suggested that the dietary replacement of FM by GCM could replace up to 164 g/kg without negatively affecting growth, feed utilization, and improved blood hematology and serum proteins of Nile tilapia, even if mild histomorphological changes of hepatocytes were observed. In addition, replacing FM with GCM111 obtained the highest growth performance without any negative effects to Nile tilapia.

Zinc-aspirin preconditioning reduces endothelial damage of arterial grafts in a rodent model of revascularization.

Coronary artery bypass grafting (CABG) is the most common cardiac surgical procedure. The prognosis of revascularization via CABG is determined by the patency of the used grafts, for which an intact endothelium is essential. The degree of ischemia-reperfusion injury (IRI), which occurs during the harvest and implantation of the grafts, is an important determinant of graft patency. Preconditioning with aspirin, a nonsteroidal anti-inflammatory drug has been shown to reduce the functional and molecular damage of arterial grafts in a rodent model. Studies have found that the zinc-aspirin complex may be able to exert an even better protective effect in pathological cardiovascular conditions. Thus, our aim was to characterize the protective effect of zinc-aspirin complex on free arterial grafts in a rodent model of revascularization. Donor Lewis rats were treated with either zinc-aspirin, aspirin, or placebo (n = 8) for 5 days, then the aortic arches were harvested and stored in cold preservation solution and implanted heterotopically in the abdominal cavity of the recipient rats, followed by 2 h of reperfusion. There was also a non-ischemia-reperfusion control group (n = 8). Functional measurements using organ bath and histomorphological changes using immunohistochemistry were analyzed. The endothelium dependent maximal vasorelaxation was improved (non-transplanted control group: 82% ± 3%, transplanted control group: 14% ± 2%, aspirin group: 31% ± 4%, zinc-aspirin group: 52% ± 4%), the nitro-oxidative stress and cell apoptosis decreased, and significant endothelial protection was shown in the groups preconditioned with aspirin or zinc-aspirin. However, zinc-aspirin proved to be more effective in the reduction of IRI, than aspirin alone. Preconditioning with zinc-aspirin could be a promising way to protect the function and structural integrity of free arterial grafts, thus improving the outcomes of CABG.

Histomorphological changes and alterations in the reproductive parameters of male wistar rats following exposure to ethanolic leaf extract of Datura stramonium (GEGEMU)

Histomorphological changes and alterations in the reproductive parameters of male wistar rats following exposure to ethanolic leaf extract of Datura stramonium (GEGEMU)