Temporal profiling of physiological, histological, and transcriptomic dissection during auxin-induced adventitious root formation in tetraploid Robinia pseudoacacia micro-cuttings.

MAIN CONCLUSION: Optimal levels of indole-3-butyric acid (IBA) applied at the stem base promote adventitious root (AR) initiation and primordia formation, thus promoting the rooting of leafy micro-cuttings of tetraploid Robinia pseudoacacia. Tetraploid Robinia pseudoacacia L. is a widely cultivated tree in most regions of China that has a hard-rooting capability, propagated by stem cuttings. This study utilizes histological, physiological, and transcriptomic approaches to explore how root primordia are induced after indole butyric acid (IBA) treatment of micro-cuttings. IBA application promoted cell divisions in some cells within the vasculature, showing subcellular features associated with adventitious root (AR) founder cells. The anatomical structure explicitly showed that AR initiated from the cambium layer and instigate the inducible development of AR primordia. Meanwhile, the hormone data showed that similar to that of indole-3-acetic acid, the contents of trans-zeatin and abscisic acid peaked at early stages of AR formation and increased gradually in primordia formation across the subsequent stages, suggesting their indispensable roles in AR induction. On the contrary, 24-epibrassinolide roughly maintained at extremely high levels during primordium initiation thoroughly, indicating its presence was involved in cell-specific reorganization during AR development. Furthermore, antioxidant activities transiently increased in the basal region of micro-cuttings and may serve as biochemical indicators for distinct rooting phases, potentially aiding in AR formation. Transcriptomic analysis during the early stages of root formation shows significant downregulation of the abscisic acid and jasmonate signaling pathways, while ethylene and cytokinin signaling seems upregulated. Network analysis of genes involved in carbon metabolism and photosynthesis indicates that the basal region of the micro-cuttings undergoes rapid reprogramming, which results in the breakdown of sugars into pyruvate. This pyruvate is then utilized to fuel the tricarboxylic acid cycle, thereby sustaining growth through aerobic respiration. Collectively, our findings provide a time-course morphophysiological dissection and also suggest the regulatory role of a conserved auxin module in AR development in these species.

Bone microstructure as an indicator of digging ability in moles (Talpidae, Eulipotyphla).

Talpid moles (Talpidae, Eulipotyphla) are mammals highly specialised in burrowing using their forelimbs. Fossoriality has allowed moles to expand their ecological niche by enabling access to subterranean resources and spaces. This specialisation in burrowing has led to adaptations in the forelimb bones of moles for humeral rotation digging, a distinctive strategy unparalleled among other diggers. While bone robustness has been examined in moles through external morphology, the adaptation of bone microstructure to digging strategy remains unclear. Based on two assumptions, (1) the humerus of moles is subjected to a torsional load due to humeral rotation digging, and (2) the magnitude of torsional load correlates with the compactness of the substrate in which the individuals can dig, we hypothesised that humeral rotation digging influences bone microstructure. Comparative analyses of transverse sections from the humeri and femora of three mole species (Mogera imaizumii, Mogera wogura and Urotrichus talpoides; Talpidae) and an outgroup eulipotyphlan (Suncus murinus; Soricidae) revealed that (1) vascular canals distributed in the humeri of moles align more predominantly circumferential along the bone walls, indicating an adaptation to the torsion generated by humeral rotation digging, and (2) the laminarity of vascular canals, particularly in Mogera species compared with Urotrichus, potentially reflects differences in the magnitude of load due to substrate compactness during digging. The aligned vascular canals are distinctive traits not observed in mammals employing other digging strategies. This suggests that vascular canal laminarity can be an indicator of not only humeral rotation digging in fossorial animals, but also the variation of eco-spaces in talpid species.

Salivary duct carcinoma with squamous differentiation: histomorphological and immunophenotypical analysis of six cases.

BACKGROUND AND AIMS: Salivary duct carcinoma (SDC) is an aggressive salivary malignancy with multiple morphological subtypes. Primary salivary squamous cell carcinoma (SCC) requires exclusion of high-grade salivary malignancies and metastatic disease and is considered exceptionally rare. We report six cases of SDC with resemblance to SCC on account of variable, but often extensive, squamous differentiation. METHODS AND RESULTS: A retrospective review (2009-2023) at two institutions of SDC with histological and immunophenotypical evidence of squamous differentiation identified six cases. Medical charts and available glass slides were reviewed. There were five males and one female with a median age of 63 years, with tumours involving the parotid (five of six) and submandibular (one of six) glands. All six tumours showed a conventional SDC component comprising < 5-90% of viable tumour. Squamous differentiation comprised 10-95%+ (> 75% in three of six cases) of total viable tumour, and demonstrated CK5/6, p63 and/or p40 immunoexpression in all cases. A sarcomatoid component, comprising 10-60% of viable tumour, was present in three of six (50%) cases. All tumours were androgen receptor (AR)-positive, but only two of six (33.3%) retained AR immunoreactivity in the squamous component. Metastatic SDC to regional lymph nodes exhibited exclusive squamous differentiation in two of six (33.3%) cases. CONCLUSION: Squamous differentiation, histologically and immunophenotypically, can be extensive in SDC. AR expression may be lost in the squamous component and metastases may demonstrate only squamous differentiation. These findings cast further doubt on the existence of primary salivary SCC. SDC should be considered whenever encountering a carcinoma with squamous differentiation in major salivary glands or within cervical lymph nodes in the setting of a salivary mass.

Dietary Pediastrum boryanum microalgal extract improves growth, enhances immunity, and regulates immune-related genes in Nile tilapia.

BACKGROUND: Identifying alternative sustainable feed sources with high nutritional values is crucial for the future of environmentally and socially responsible aquaculture. In this regard, microalgae have been proven to have positive effects on fish health, which overwhelmed our interest in this study. METHODS: Pediastrum boryanum (P. boryanum) was incorporated into Nile tilapia feed at concentrations of 0, 0.75, and 1.5 mg/kg, as control, PbExt0.75, and PbExt1.5 groups to assess its effects on growth and biochemical indices, oxidant/antioxidant activities, immune and stress-related gene expression, and intestinal morphology. RESULTS: After 8 weeks, fish fed P. boryanum supplemented feed exhibited significant increases in final weight, length, condition factor, body weight gain, and specific growth rate, while the spleen-somatic index (SSI) and hepatosomatic index (HSI) showed no significant differences compared to the control group. Dietary P. boryanum supplementation also enhanced IgM levels and lysozyme activity, along with no marked effect on markers of liver function enzymes (alanine aminotransferase/ALT and aspartate aminotransferase/AST) or protein status (total protein and albumin). Furthermore, P. boryanum addition increased the activity of superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) enzymes, highlighting its antioxidant potential, whereas malondialdehyde (MDA) concentrations showed no significant differences among the groups. Gene expression analysis revealed that tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and transforming growth factor-ß1 (TGF-ß1) expression notably increased in groups fed P. boryanum containing feed, while no significant difference was observed in hepatic Heat Shock Protein 70 (HSP70) mRNA expression. Histopathological examination revealed no adverse effects of P. boryanum supplementation on the liver, spleen, or intestinal tissues. Villous height and villous surface area were notably increased in the high P. boryanum supplementation group, suggesting improved intestinal integrity and nutrient absorption. CONCLUSION: Dietary P. boryanum supplementation can potentially improve growth performance, immune response, antioxidant status, and intestinal health of Nile tilapia, making it a promising candidate for sustainable aquaculture.

Effects of graded levels of dietary microbial 6-phytase on performance, intestinal histomorphology, caecal microbial population and short-chain fatty acid composition of Lohmann white-classics.

1. This study was conducted to determine the effects of graded levels of phytase on the performance, egg quality and gut health of white laying hens.2. Treatments consisted of a negative control (NC) diet containing 0.14% available phosphorus (avP), positive control (PC) diet containing 0.35% avP provided via dicalcium phosphate (DCP) and DCP replaced in the PC by with three graded levels of phytase derived from Komagataella phaffii at 500 (PC-500), 750 (PC-750) and 1000 (PC-1000) FTU/kg which provided 0.176%, 0.188% and 0.200% of avP, respectively.3. Egg production, feed intake, feed conversion ratio and jejunal morphometry were negatively affected in NC-fed birds (p < 0.05). Considering the whole period, birds fed a diet supplemented with graded levels of phytase shared the same egg production and feed intake levels with PC birds (p < 0.05). Feed conversion ratio was significantly lowered by 4.9%, 1.6% and 7.6% in hens fed on diets PC-500, PC-750 and PC-1000, respectively compared to those fed the PC (p < 0.05).4. Neither of the dietary treatments affected cracked eggs, dirty eggs, eggshell breaking strength and eggshell thickness. Dietary supplementation of phytase significantly increased villus surface area by 15%, 36% and 40% in PC-500, PC-750 and PC-1000 birds, respectively compared to PC (p < 0.05).5. A significant increase in lactobacillus count was observed in line with increasing the level of phytase (p < 0.05). Dietary treatments had no effect on the caecal coliform or aerobic populations. Furthermore, phytase supplementation significantly increased the concentrations of total caecal short-chain fatty acid (SCFA; p < 0.01).6. In conclusion, along with improving performance parameters, the inclusion of phytase in laying hen diets can ameliorate intestinal morphology and stimulate caecal microflora and increase SCFA concentrations.

Physiological and immunological aspects of feed restriction and its beneficial impacts in fattening rabbits' productivity-an updated review.

Two feed restriction (FR) regimens are utilized with weaned rabbits including a quantitative FR (amount-limited feed or time-restricted admission to feed) and a qualitative FR (modulation of diet content especially protein and energy). The use of post-weaning FR may help in preventing post-weaning digestive disorders, stimulating compensating growth, improving feed efficiency, and decreasing carcass fat content. Interestingly, FR may contribute to changing meat's chemical composition and its physical quality attributes by regulating the morphological and biochemical characteristics of muscle fibers. Also, FR could enhance the gastrointestinal tract development, its histomorphology, and improve feed digestibility and absorption. Furthermore, FR regimens are involved in establishing gut microbial balance and enhancing the host immunological response. It might be concluded that post-weaning FR is involved in influencing the physiological and immunological aspects of growing rabbits. It might be documented that light to mild FR (i.e., 80-90% AL), early (i.e., at the first 2 weeks post-weaning), and relatively short in duration (i.e., for 2-3 weeks) had no negative effects on live body weight, while severing FR reduced live body weight in comparison with ad libitum rabbits.

Molecular mechanism of somatic embryogenesis in paeonia ostii 'Fengdan' based on transcriptome analysis combined histomorphological observation and metabolite determination.

BACKGROUND: Tree peony (Paeonia sect. Moutan DC.) is a famous flower native to China with high ornamental, medicinal, and oil value. However, the low regeneration rate of callus is one of the main constraints for the establishment of a genetic transformation system in tree peony. By histomorphological observation, transcriptomic analysis and metabolite determination, we investigated the molecular mechanism of somatic embryogenesis after the establishment of a culture system and the induction of somatic embryo(SE) formation. RESULTS: We found that SE formation was successfully induced when cotyledons were used as explants. A total of 3185 differentially expressed genes were screened by comparative transcriptomic analysis of embryogenic callus (EC), SE, and non-embryogenic callus (NEC). Compared to NEC, the auxin synthesis-related genes GH3.6 and PCO2 were up-regulated, whereas cytokinin dehydrogenase (CKX6) and CYP450 family genes were down-regulated in somatic embryogenesis. In SE, the auxin content was significantly higher than the cytokinin content. The methyltransferase-related gene S-adenosylmethionine synthase (SAMS) and the flavonoid biosynthesis-related gene (ANS and F3'5'H) were down-regulated in somatic embryogenesis. The determination of flavonoids showed that rhoifolin and hyperoside had the highest content in SE. The results of transcriptome analysis were consistent with the relative expression of 8 candidate genes by quantitative polymerase chain reaction analysis. CONCLUSION: The results revealed that auxin and cytokinin may play a key role in 'Fengdan' somatic embryogenesis. The genes related to somatic embryogenesis were revealed, which has partly elucidated the molecular mechanism of somatic embryogenesis in 'Fengdan'.

Fin systems comparative anatomy in model Batoidea Raja asterias and Torpedo marmorata: Insights and relatioships between musculo-skeletal layout, locomotion and morphology.

The macroscopic and microscopic morphology of the appendicular skeleton was studied in the two species Raja asterias (order Rajiformes) and Torpedo marmorata (Order Torpediniformes), comparing the organization and structural layout of pectoral, pelvic, and tail fin systems. The shape, surface area and portance of the T. marmorata pectoral fin system (hydrodynamic lift) were conditioned by the presence of the two electric organs in the disk central part, which reduced the pectoral fin surface area, suggesting a lower efficiency of the "flapping effectors" than those of R. asterias. Otherwise, radials' rays alignment, morphology and calcification pattern showed in both species the same structural layout characterized in the fin medial zone by stiffly paired columns of calcified tiles in the perpendicular plane to the flat batoid body, then revolving and in the horizontal plane to continue as separate mono-columnar rays in the fin lateral zone with a morphology suggesting fin stiffness variance between medial/lateral zone. Pelvic fins morphology was alike in the two species, however with different calcified tiles patterns of the 1st compound radial and pterygia in respect to the fin-rays articulating perpendicularly to the latter, whose tile rows lay-out was also different from that of the pectoral fins radials. The T. marmorata tail-caudal fin showed a muscular and connective scaffold capable of a significant oscillatory forward thrust. On the contrary, the R. asterias dorsal tail fins were stiffened by a scaffold of radials-like calcified segments. Histomorphology, heat-deproteination technique and morphometry provided new data on the wing-fins structural layout which can be correlated to the mechanics of the Batoid swimming behavior and suggested a cartilage-calcification process combining interstitial cartilage growth (as that of all vertebrates anlagen) and a mineral deposition with accretion of individual centers (the tiles). The resulting layout showed scattered zones of un-mineralized matrix within the calcified mass and a less compact texture of the matrix calcified fibers suggesting a possible way of fluid diffusion throughout the mineralized tissue. These observations could explain the survival of the embedded chondrocytes in absence of a canalicular system as that of the cortical bone.

The occurrence, progression and development of four types of gastric mucosal atrophic lesions and their histopathological characteristics.

OBJECTIVE: To investigate the occurrence and development of gastric mucosal atrophic lesions and their histopathological characteristics. METHODS: Histopathological diagnosis and immunohistochemical staining using the EnVision two-step method were conducted on 1969 gastric mucosal atrophic lesions obtained from gastroscopic biopsy specimens. A total of 48-month three-stage endoscopic biopsy follow-ups were performed. RESULTS: When the gastric mucosal epithelium was affected by infection, chemical irritation, or immune or genetic factors, the gastric mucosal epithelium glands atrophied, the mucosa became thinner, the number of glands decreased, the intestinal epithelium progressed to metaplasia and smooth muscle fibre became hyperplasia. Such changes may lead to the proliferation and dysplasia of epithelial cells of the gastric mucosa and neoplastic hyperplasia in nature; this is referred to as gastric mucosal atrophic lesions in this study. According to this definition, the present study divided gastric mucosal atrophy into four types: (1) glandular atrophy of the lamina propria; (2) compensatory proliferative atrophy; (3) intestinal metaplasia atrophy; and (4) smooth muscle proliferative atrophy. The incidence rates of the above were 40.1% (789/1969), 14.3% (281/1969), 27.8% (547/1969) and 17.9% (352/1969), respectively. One- to 4-year follow-ups found that the changes were not significant and that the percentages of patients with disease exacerbation were 85.7% (1688/1969) and 9.8% (192/1969). The percentages of patients who developed low-grade intraepithelial neoplasia and high-grade intraepithelial neoplasia were 2.8% (55/1969) and 1.1% (21/1969), respectively; 0.7% (13/1969) of patients developed intramucosal cancer. CONCLUSION: Gastric mucosal atrophic lesions and histopathological staging are based on the morphological characteristics of gastric mucosal atrophy and the hypothesis of malignant transformation of cells during the occurrence and development of mucosal atrophy. Mastering pathological staging is beneficial to clinicians for enacting precise treatment and is important for reducing the incidence of gastric cancer.

Nutritional and immunological evaluation of Nannochloropsis oculata as a potential Nile tilapia-aquafeed supplement.

Nannochloropsis oculata (N. oculata) is a marine microalga containing bioactive compounds and a high omega-3 polyunsaturated fatty acid (ω-3 PUFAs). Therefore, it is very promising for nutraceutical and the functional food industry applications. Three groups of Nile tilapia (forty-five fish/group) were fed on basal diets or diets containing 5% (N5) or 10% (N10) of the microalga N. oculata for seven weeks. Fish growth performance, proximate composition, and lipid (fatty acids/ FAs and lipoproteins) profile were estimated. In addition, the expression pattern of some lipid metabolism and immune-relevant genes were assessed. An enhancement in whole body crude protein and growth indices of Nile tilapia was observed on both the supplemented groups N5 and N10. Higher levels of high-density lipoproteins (HDL); and lower levels of the low-density lipoproteins (LDL) were evident in both supplemented groups, while the cholesterol and triglycerides (TG) levels were similar among groups. Ω-3 PUFAs were the significant FAs profile of tilapia fed on N. oculata-supplemented diets in terms of eicosapentaenoic acid, docosahexaenoic acid, and n3/n6 ratio. Concerning the gene expression pattern, heat-shock protein70, glutathione-S-transferase, glutathione peroxidase, and interleukin-1ß (IL-1ß) were elevated significantly in both supplemented groups. IL-10 is only upregulated in the N10 group. The lipid metabolism-related gene expression showed downregulation of only fatty acid synthase (FAS) in both supplemented groups, with no statistical changes in Peroxisome proliferator-activated receptor alpha (PPARα). Tumor necrosis factor-α (TNF-α), Transforming growth factor-ß1 (TGF-ß1), and the apoptotic related genes [caspase3 and Proliferating cell nuclear antigen (PCNA)] showed insignificant changes among groups. The histopathological examination of the intestine, liver, and spleen supports our findings and confirms the benefits and safeness of N. oculata dietary inclusion. Collectively, N. oculata is a very promising nutraceutical for improving fish health and sustainability of aquaculture production.

Gastrointestinal permeability and inflammatory status of preweaning dairy calves in response to decreasing the ratio of n-6 to n-3 fatty acid in milk replacer.

The ratio of n-6 to n-3 fatty acid (FA) is between 2 and 10 times higher in milk replacer (MR) than in whole milk, which may promote inflammation and compromise the integrity of the intestinal epithelium. To evaluate how decreasing the n-6:n-3 FA ratio of MR affects gastrointestinal (GIT) permeability and inflammatory status, 30 dairy calves (2.8 ± 1.06 d of age; mean ± standard deviation) were randomly assigned to be fed an MR with an n-6:n-3 FA ratio of 40:1 (CON; 29.3% crude fat of DM; n = 15) or 6.5:1 (n-3; 29.1% crude fat of DM; n = 15). Calves were fed 7.0 L/d in 2 meals. Calves were weighed and fecal consistency was analyzed weekly. On d 22, calves were administered Cr-EDTA, lactulose, and d-mannitol to assess GIT permeability. Blood and total urine were sequentially collected for 6 and 24 h, respectively, and analyzed for marker content. Whole blood collected 4 h after the meal was subjected to an ex vivo lipopolysaccharide (LPS) challenge to evaluate cytokine secretion from blood cells. Calves were euthanized on d 25 for collection of intestinal tissue samples. Tissue samples were processed to assess FA composition by gas chromatography, histomorphology by bright-field microscopy, and gene expression of tight junction proteins, lipid metabolism enzymes, and immune molecules by real-time quantitative PCR. Data were analyzed using PROC GLIMMIX in SAS (version 9.4, SAS Institute Inc.). Growth performance and fecal consistency were unaffected. Calves fed MR with a lower ratio of n-6 to n-3 FA had 2-fold higher n-3 FA contents and 2-fold lower ratios of n-6 to n-3 FA in proximal jejunum and ileum tissues. Total urinary recovery (0-24 h relative to marker administration) and plasma concentrations of the markers were unaffected. Expression of TJP1 tended to be higher in proximal jejunum tissue and lower in ileum tissue of n-3 calves. The expression of TLR4 and TNFA tended to be higher and CD14 was higher in ileum tissue of n-3 calves. Plasma concentrations of interleukin-4 were decreased in response to the ex vivo LPS challenge in n-3 calves. Histomorphology and GIT permeability were largely unaffected by treatment. Furthermore, the inclusion of linseed and algae oil may promote inflammation, as suggested by greater concentrations of the acute-phase proteins haptoglobin and serum amyloid A postprandially, demonstrating that fat sources should be evaluated for their suitability for MR formulations. Understanding how MR composition affects dairy calf health may improve nutritional strategies on farm.

Antireproductive changes instigated by efficient drug delivery via papaya seed chloroform extract-based nanoparticles in male rat Bandicota bengalensis.

Anti-reproductive potential of papaya seed chloroform extract-based solid lipid nanoparticles (PSCEN) was investigated for the first time in lesser bandicoot rat, Bandicota bengalensis. Mature male rats (n = 30 per group) were fed bait (loose mixture of cracked wheat, powdered sugar, and groundnut oil in the ratio 88:10:2) containing two different concentrations of PSCEN (5% and 10%) in a bi-choice condition for 15 days with one group as vehicle control. The ingestion of active ingredient in 15 days treatment was significantly (P ≤ 0.05) higher by rats treated with 10% PSCEN (39.17-58.70 g/kg body weight) as compared to rats treated with 5% PSCEN (21.30-33.23 g/kg body weight). A dose dependent significant (P ≤ 0.05) decrease was observed in the level of testosterone, FSH, LH and GnRH in plasma of treated rats. A significant (P ≤ 0.05) decrease was also observed in level of total soluble proteins, total lipids, phospholipids and cholesterol in both plasma and testicular tissue, and level of 17ß-HSD and 3ß-HSD in testicular tissue indicating anti-reproductive effects of PSCEN treatment. There was observed significant (P ≤ 0.05) effect of treatment on histomorphology of testis and cauda epididymis in the form of reduced tubular diameter, germinal epithelial thickness, number of germ cells and dissociation of epithelial cycle in seminiferous tubules, and reduced tubular diameter, increased epithelial thickness, vacuolization, loose contact of principle cells and reduced number of spermatozoa in the cauda epididymal tubules. Maximum antifertility effect was observed with 10% PSCEN treatment, which was not reversed upto 105 days of treatment withdrawal indicating long-term efficacy. The current investigation suggests the use of PSCEN in the management of reproduction of B. bengalensis by exerting influence on testicular and cauda epididymal functions and biochemical parameters.

A case of racemiform trichoblastoma in a dog.

Trichoblastoma, which is common in dogs, is now occurring with other cellular changes outside the recognized forms to warrant their continuous evaluation for proper elucidation even as their causes largely remain unknown. A case at hand involved a 9-year-old Caucasian dog, which weighed 35 kg with chief complaint of a progressive bleeding mass on the scalp. The dog had an up-to-date vaccination record and all vital parameters were within optimum ranges. The surgical excision of the firm, solitary, and alopecic mass with traumatized upper surface revealed the presence of a well-demarcated and unencapsulated mass composed of grapes-like nests of basaloid epithelial cells within follicular stroma devoid of stromal necrosis, inflammatory cellular infiltration, and neoplastic epidermal connection. However, there was tissue necrosis, hemorrhages, and inflammatory cellular infiltrates on the exposed upper part of the traumatized growth. Immunohistochemical analysis showed positive reactivity to AE1/AE3, CK5/6, and p63 but negative immunoreactivity to CK7, CK20, CEA, and TTF-1. The histomorphological and immunohistochemical evaluation of the mass on the scalp of the dog suggested a solitary racemiform trichoblastoma with a traumatized exposed upper part despite basal cell carcinoma mimicry where histological diagnosis currently hold sway over immunohistochemical evaluation.

Melatonin ameliorates osteoarthritis rat cartilage injury by inhibiting matrix metalloproteinases and JAK2/STAT3 signaling pathway.

OBJECTIVE: To observe the effect of melatonin intervention on rat knee osteoarthritis (KOA) model and explore its mechanism. METHODS: A total of 81 Sprague-Dawley (SD) rats were employed. Haematoxylin and eosin (H&E) staining and safranin o-solid green staining were used to observe the changes of pathology in KOA, and inflammation factors in serum were detected by enzyme-linked immunosorbent assay (ELISA), type II collagen (Col-II) was detected by immunohistochemistry, chondrocyte apoptosis was detected by TdT-mediated dUTP nick-end labeling (TUNEL). The expression of matrix metalloproteinases (MMPs) and JAK2/STAT3 signaling were detected by western blot. RESULTS: Melatonin treatment ameliorated the histomorphology of knee joint in rats compared to the model group. The contents of TNF-α, IL-6, and IL-1ß in serum were decreased after melatonin treatment. In addition, compared to the model group, the positive expression of Col-II increased, the chondrocyte apoptosis decreased after melatonin treatment. Interestingly, the expression levels of MMP3, MMP9, MMP13, p-JAK2 and p-STAT3 decreased (p < 0.05). Importantly, melatonin combined with AG490 is significantly ameliorates histomorphology of knee joint, reduced cartilage loss compared with melatonin treatment alone. CONCLUSIONS: Melatonin treatment can effectively diminish the cartilage injury. Its mechanism may be related to protect the articular cartilage by reducing the release of inflammatory factors, inhibit the expression of MMPs and JAK2/STAT3 signaling.

Exogenous protease influences protein digestibility, growth performance, and gut microflora in weanling pigs on a limited protein diet.

The study was conducted to evaluate the impact of dietary level of crude protein (CP) and protease supplementation on growth performance, digestibility of nutrients, intestinal morphology, and gut microbiota in weaning pigs. Three hundred cross-bred piglets (Duroc × Landrace × Yorkshire) were allotted to five dietary treatments on the basis of initial body weight (BW) and sex. Pigs were group-housed in pens with each treatment with 10 replicate pens with six pigs per pen. The treatments included a standard diet (STD), STD with 0.6% lower protein (STD0.6), STD with 0.6% lower protein and protease supplementation (Pro0.6), STD with 1.0% lower protein (STD1.0), STD with 1.0% lower protein and protease supplementation (Pro1.0). Results indicated a higher BW (p < 0.05) of piglets in the Pro0.6 group at days 0-42 compared to the STD0.6 and STD1.0 groups. The average daily gain was higher (p < 0.05) in the Pro0.6 treatments at days 0-42 compared to the STD0.6 and STD1.0. The gain to feed ratio was higher (p < 0.05) in the STD, and Pro0.6 groups compared to the STD0.6, Pro1.0 and the STD1.0 groups at days 0-42. Dry matter digestibility was lower (p < 0.05) in the STD1.0 group than the Pro0.6 and Pro1.0 groups. The crude protein digestibility was higher (p < 0.05) in the Pro0.6 group compared to the STD, STD0.6 and STD1.0 treatment groups while crude fat digestibility was higher (p < 0.05) in the STD and Pro0.6 compared with the STD0.6 and STD1.0 groups. Digestibility was higher for histidine (p < 0.05), leucine (p < 0.05) in the protease Pro0.6 and Pro1.0 groups than in the STD0.6 and STD1.0 groups. The digestibility of non-essential AA was higher for alanine (p < 0.05) in the Pro0.6 than the STD1.0 group. For faecal microbial population, Faecalibacterium abundance was higher (p < 0.05) in the Pro0.6 compared to all the other groups while the population of Actinobacteria was greater (p < 0.05) in the STD group and lowest in the Pro1.0 treatment. In the ileum, villus height was greater (p < 0.05) in the protease Pro0.6, and Pro1.0 groups compared to the STD0.6, and STD1.0 groups while the villus height to crypts depth ratio was lower (p < 0.05) in the STD 1.0 group compared to the STD, Pro0.6, and Pro1.0 groups. Based on these results, dietary protease supplementation improved nutrient digestibility and gut histo-morphology translating to improved utilisation of nutrients thus positively impacting growth performance in weaned pigs. Further, reducing the CP content in the diets increased the abundance of Muribaculaceae while protease supplementation increased the population of Faecalibacterium in the gut of the weanling piglets on the STD0.6 diet.

Efficacy of mono- and multistrain synbiotics supplementation in modifying performance, caecal fermentation, intestinal health, meat and bone quality, and some blood biochemical indices in broilers.

The superiority of synbiotics in terms of their biological effects depends primarily on a suitable combination of both components, pro-biotic and pre-biotic. The present study was conducted to compare the efficacy of mono- and multistrain synbiotics on overall performance, caecal fermentation, intestinal health, meat and bone quality along with some blood biochemical indices in broilers. A total of 231, 1-day-old male Ross 308, broiler chicks were randomly assigned to three experimental groups using 11 replicates each and seven chicks/replicate. The dietary treatments included control group with no synbiotic supplementation, monostrain (Maflor) and multistrain (Maflor plus) synbiotic groups with 1 g/kg of added synbiotics each. Synbiotics feeding significantly improved animal performance with a clear impact on meat quality in terms of low-fat, optimum ultimate pH24, higher water holding capacity, and lower drip and cooking losses. Of the two synbiotics, multistrain seemed to have responded better in modifying small intestinal epithelia and fermentation metabolites, although both synbiotics were comparable in reducing the pathogen load. Load-bearing capacity of both leg bones (femur and tibia) was also enhanced with synbiotics supplementation, which was also reflected in their mineral profile. The blood serum biochemical analysis showed a reduction in circulating cholesterol and triglycerides levels and an increment in IgA and IgG concentrations. In conclusion, the remarkable efficacy of tested synbiotics in providing higher growth, better meat quality in tandem with the optimum gut environment, lower pathogen load, healthy epithelia, immunomodulation, hypocholesterolemic, and hypotriglyceridemic effects affirms their great potential to be used as feed additives in broiler diets. Contrary to our expectations, the effectiveness of a mono-strain in comparison to multistrain synbiotic in improving almost all the features investigated was also notable. Further evaluation under challenging conditions should be explicitly conducted to achieve more comprehensive results.

Effects of supplementing organic- and inorganic-based selenium with vitamin E on intestinal histomorphology, caecal bacterial proliferation, and short-chain fatty acid profile in layer hens.

The effects of supplementing different dietary selenium (Se) sources on intestinal histomorphology, caecal bacterial proliferation, and caecum short-chain fatty acid in layer chickens were studied. A total of 120 of 54-week-old Lohman Brown Classic laying hens were subjected to four dietary treatments: control group with no Se supplementation (T1), 0.3 mg/kg of sodium selenite (T2), 0.3 mg/kg of selenium yeast (T3), and 0.3 mg/kg of bacterial Se (Stenotrophomonas maltophilia, ADS18) (T4). All treatments were also supplemented with 250 mg/kg of vitamin E. Results showed significantly (p < 0.05) higher villus height in different small intestine parts in hens fed with diets containing organic Se sources (T3 and T4), as compared to inorganic sources (T2) and control (T1). A greater crypt depth was observed for the T1 group, as compared to T3 and T4 groups. The total bacterial populations of Bifidobacteria spp. and Lactobacilli spp. were significantly increased (p < 0.05), and tEscherichia coli population was significantly decreased (p < 0.05) in T3 and T4 groups. No effect on the total caecal short-chain fatty acid (SCFAs) content was observed. However, there was a significant increase (21.5%) of the butyric acid content in T4 group. In conclusion, organic Se supplementation, particularly bacterial organic Se, enhances intestinal histomorphology, increases the beneficial caecal bacterial proliferation, and increases butyric acid content.

A Comparative Study of Bone Tissue Morphology and Bone Turnover Markers in Different Stages of Kümmell's Disease.

Although the incidence of Kümmell's disease (KD) has increased significantly in recent years, its pathological mechanism is still unclear. The aim of this study is to investigate the histomorphological characteristics and the kinetics of bone turnover markers following KD. This study involves 82 patients with KD, and fasting blood samples were collected to detect the serum concentration of bone turnover markers. A transpedicular bone biopsy was performed to collect bone biopsy specimens during vertebroplasty surgery. According to Li's staging system for KD, all cases were divided into 3 stages. Comparisons of the 3 stages and their kinetics were conducted. 19 (23.2%) patients were classified as Stage I, 39 (47.5%) as Stage II, 24 (29.2%) as Stage III. Bone histomorphological analysis showed that the ratios of WBV and FV reached a peak value (14.23 ± 0.62 and 54.63% ± 3.52%; p = 0.001 and 0.001) at Stage I. The ratios of NBV remained low (4.81% ± 2.61%) in Stage I, but reached a peak value (18.50% ± 2.77%; p = 0.001) in Stage III. Bone metabolism index level showed that the serum concentration of OST and ß-CTX continued to rise after fracture, reaching a peak value of (38.15 ± 3.84 and 1.31 ± 0.16 ng/Ml; p = 0.073 and 0.026), while PINP reached its valley value (48.57 ± 7.25 ng/Ml; p = 0.069) in Stage III. A significant and negative correlation was found between the ratio of ß-CTX and EBV/TV (p= 0.0194, r = -0.2037), and FV/TV (p= 0.0001, r = -0.5368). At the same time, ß-CTX had a positive significant correlation with the NBV/TV (p= 0.0001, r = 0.6218). Bone histomorphometric analysis and bone turnover markers showed that KD has a possibility of healing in the early stage.

Cytology of angiofibroma of soft tissue of the inguinal region.

Angiofibroma of the soft tissue is a recently described benign fibroblastic/myofibroblastic tumour. This report describes the cytology of an angiofibroma of soft tissue occurring in a 30-year-old lady which showed bland spindle cells, occasional polygonal cells with nuclear grooving, prominent vessels, frayed stroma around the blood vessels, and scattered lymphocyte-rich inflammatory cells in the background.

Type 2 diabetes and the clinically normal pulp: An in vitro study.

AIM: The aim of this study was to investigate the effect of type 2 diabetes (T2D) on clinically normal dental pulp tissue by using special stains and immunohistochemistry (IHC) to determine the morphology of the coronal pulp and distribution of immune markers in non-T2D and T2D groups. METHODOLOGY: Ethics approval for this in vitro pilot study was obtained from the University of Otago Human Ethics Committee (16/069). Twenty extracted permanent molar teeth diagnosed as having clinically normal pulp status were collected. Ten teeth were from participants with well-controlled T2D and ten from participants without diabetes (non-T2D). Each tooth was sectioned transversely at the cemento-enamel junction before the crowns were decalcified and embedded in paraffin. Sections were stained with haematoxylin and eosin, Massons trichrome, and van Gieson stains for histological and morphological evaluation. IHC using anti-CD4, anti-CD68 and anti-CD83 and anti-IL1ß, anti-IL6, anti-IL17, anti-TNF-α, anti-TLR2, anti-TLR4 and anti-FOXP3 identified proteins of interest. Qualitative and semi-quantitative analyses evaluated the morphology of the dental pulp and protein expression. Data analyses were performed with GraphPad Prism, using Student's t-test and multiple regression using SPSS at p < .05. RESULTS: Special stains demonstrated morphological differences in the T2D dental pulp compared with non-T2D. Qualitative analysis indicated that the pulp in the T2D samples was consistently less cellular, less vascular, showed evidence of thickened blood vessel walls, increased pulp calcification and collagen deposition. Semi-quantitative analysis of IHC samples showed the T2D pulp had significantly increased expression of macrophage and dendritic cell markers CD68 (p < .001) and CD83 (p = .04), and there was significantly greater expression of inflammatory cytokines IL1ß (p = .01), IL6 (p < .0001), IL17 (p < .0001) and TNF-α (p = .01). T2D samples showed a significant increase in markers of innate inflammation, TLR2 (p < .001) and TLR4 (p < .001) and decreased expression of regulatory T-cell marker, FOXP3 (p = .01). Multiple regression showed that age-corrected differences were statistically significant. CONCLUSION: Preliminary findings suggest that T2D may exert a similar response in the pulp to complications in other body sites. Hyperglycaemia is associated with changes in the morphology of the clinically normal dental pulp with altered immune cell and cytokine expression.