SYN Group Research Articles

Gastrointestinal, Behaviour and Anxiety Outcomes in Autistic Children Following an Open Label, Randomised Pilot Study of Synbiotics vs Synbiotics and Gut-Directed Hypnotherapy.

Alterations of the microbiome-gut-brain (MGB) axis have been associated with autism spectrum disorder (ASD) and disorders of gut-brain interaction (DGBI). DGBI are highly prevalent in autistic children and are associated with worsening behaviour and anxiety. Treatments such as probiotics, prebiotics and gut-directed hypnotherapy (GDH) have shown efficacy in improving gut symptoms in children. The primary objective of the study was to compare changes in gastrointestinal (GI) scores following a 12-week intervention of synbiotics (prebiotic + probiotic) +/- GDH with a follow-up at 24 weeks. Secondary objectives included changes in behavioural and anxiety symptoms, while changes in gut microbiome composition were assessed as an exploratory objective. Children diagnosed with ASD aged 5.00-10.99 years (n = 40) were recruited and randomised (1:1) to a 12-week intervention of either synbiotics (SYN group) or synbiotics + GDH (COM group). Both the SYN and COM group experienced significant reductions in total GI scores post-intervention and at follow-up (p < 0.001), with no superiority of the COM treatment over the SYN treatment. The COM group showed beneficial reductions in anxiety scores (p = 0.002) and irritability behaviours (p < 0.001) which were not present in the SYN group. At follow-up, only those in the COM group maintained significant reductions in GI pain scores (p < 0.001). There were significant changes in gut microbiota such as increases in Bifidobacterium animalis and Dialister in both groups over time. In conclusion,synbiotics with or without GDH may help support standard care for autistic children who suffer comorbid DGBI. The trial was prospectively registered at clinicialtrials.gov on 16 November 2020 (NCTO4639141).

Diagnostic value of the Sirtuins family in acute rejection of kidney transplantation assessed on the basis of transcriptomics and animal experiments

BackgroundThe Sirtuins (SIRT) family plays a key role in the diagnosis and treatment of many renal diseases, but no studies have been reported in acute rejection of kidney transplantation. The aim of this study was to explore the diagnostic value of SIRT family change characteristics in acute rejection of kidney transplantation. MethodsWe first explored the SIRT family expression profile in renal tissues using the HPA database; subsequently, we explored the potential biological functions and mechanistic changes during acute rejection of kidney transplantation by GSEA enrichment analysis. The Cibersort algorithm specifies the level of immune cell infiltration and explores the correlation between the SIRT family and immune cells using correlation analysis; Next, we constructed a diagnostic model using “Logistic regression analysis” and “Nomogram model”, and evaluated the diagnostic model using calibration curves and ROC curves, and the decision curve (DCA) was used to evaluate the clinical diagnostic value of SIRT family changes; Finally, we constructed a model of acute rejection of rat kidney transplantation, and assessed rat kidney function by detecting the levels of urea nitrogen and creatinine in serum. Meanwhile, the expression level of SIRT family in kidney tissues was initially verified by transcriptome sequencing and RT-PCR. ResultsWe found that all seven SIRT family members were located and expressed in renal tissues. The results of enrichment analysis revealed that a large number of immune-related biological functions and pathways are activated during acute rejection of kidney transplantation, the difference was statistically significant (p < 0.05). The Cibersort algorithm revealed significant changes in the level of infiltration of 10 immune cells (p < 0.05), while correlation analysis revealed a strong link between the SIRT family and immune cells (p < 0.05). We constructed a diagnostic model for acute rejection using seven SIRT families, and the ROC curves(AUC = 0.71)and calibration curves proved their good diagnostic value, and the DCA curves also proved the role of SIRT families in clinical decision-making. Next, we again demonstrated the good diagnostic performance of the SIRT family in ABMR and TCMR, respectively(ROC curves:AUC = 0.64，AUC = 0.81). Finally, in a rat model of acute rejection of kidney transplantation, we found that renal function (BUN and creatinine) was significantly impaired in rats in the Allo group compared to rats in the Syn group (P < 0.05). Meanwhile, by transcriptome analysis and RT-PCR assay, we found that, except for SIRT1, the remaining SIRT family members were significantly changed in kidney tissues (P < 0.05). ConclusionThe SIRT family has significant changes during acute rejection in kidney transplantation, and the SIRT family may be able to serve as a potential therapeutic target for alleviating acute rejection in kidney transplantation.

The Influence of Dietary Synbiotic on Agonistic Behavior, Stress, and Brain Monoamines via Modulation of the Microbiota–Gut–Brain Axis in Laying Hens

A complex system of neural pathways, collectively known as the microbiota–gut–brain (MGB) axis, interconnects the gut microbiota, the gastrointestinal system, and the brain along with its periphery. Previous studies have demonstrated that modulation of the MGB axis can influence stress-related behaviors such as anxiety. This connection becomes apparent in scenarios like agonistic behavior in laying hens, which is characterized by aggressive head and feather pecks, that can ultimately result in cannibalism and death. The objective was to examine the effects of a dietary synbiotic on agonistic behavior, plasma and brain monoamines, stress parameters, and cecal microbiota counts via modulation of the MGB axis. A total of 396 W36 Hy-Line laying hens were provided at random with a control (CON: basal diet) or treatment (SYN: basal diet supplemented with synbiotic) diet from 50 to 60 weeks old (nine pens/treatment, 22 birds/pen). Blood samples and video recordings (three consecutive days/week) were taken at 50 and 60 weeks. At 60 weeks, three hens/pen were euthanized for brain and cecal microbiota collection. Threatening, fighting, head, body, and feather pecking all occurred less frequently at 60 weeks in the SYN group (p &lt; 0.05). Plasma corticosterone, adrenocorticotropic hormone, dopamine, and serotonin were significantly lower while tryptophan and 5-hydroxyindoleacetic acid were significantly higher in birds from the SYN group (p &lt; 0.05). Significant differences in serotonin, 5-hydroxyindoleacetic acid, dopamine, homovanillic acid, and 3,4-dihydroxyphenylacetic acid were observed in the hypothalamus, hippocampus, and amygdala of the brain. Serotonin and dopamine turnover rates were significantly different in all three regions of the brain (p &lt; 0.05). Cecal counts of Lactobacillus and Bifidobacterium were significantly higher in the SYN group (p &lt; 0.05). Synbiotic supplementation resulted in many significant differences, indicating activation of the serotonergic systems and modulation of both the MGB axis and HPA axis with positive effects on welfare and stress.

Better clinical outcomes and faster weight bearing after medial opening-wedge high tibial osteotomy using allogeneic than synthetic graft: A secondary analysis of a Francophone Arthroscopy Society Symposium

IntroductionAlthough an autogenous graft has the highest rate of bone union to fill the void created in medial opening wedge high tibial osteotomy (MOWHTO), it also has some disadvantages, such as prolonged surgical time, donor site pain and morbidity. Two possible candidates for ideal grafts to replace autogenous grafts are allogeneic and synthetic graft, which are free from donor site pain and morbidity. However, previous reports comparing the clinical results of allogeneic to synthetic graft have been limited and controversial. The purpose of this study is to compare radiological findings and clinical outcomes of using synthetic versus allogenic graft to fill the void created in MOWHTO. HypothesisThe present clinical study hypothesized that allogenic graft to fill the void would allow the higher rate of bone union and better clinical outcomes. Material and methodsThis study compared the clinical and radiological outcomes of 95 patients who received MOWHTO to fill the void with either synthetic or allogenic graft (44 in Syn group, 51 in Allo group). Preoperatively and postoperatively, all patients were clinically evaluated; Return to work, Tegner activity score, and the Western Ontario and Macmaster University scores were reported. Radiographically, osteoarthritis grade and pre- and postoperative parameters were reported, including Hip-knee-ankle angle, mechanical lateral distal femoral angle, medial proximal tibial angle, joint line convergence angle, proximal posterior tibial angle, and limb length discrepancy. Perioperative details and complications were also reported. ResultsMean follow-up (months) were 24.0±1.3 in Syn group and 26.8±1.2 in Allo group (p=0.13). The postoperative improvement of pain and global WOMAC scores in Allo group were significantly better than in Syn group (ΔPain of WOMAC: Syn group 27.8±4.4, Allo group 49.3±3.8, p value <0.001*) (ΔGlobal score of WOMAC: Syn group 16.7±3.2, Allo group 37.4±4.9, p value=0.002*). The risk of hinge fracture in Syn group was significantly higher than in Allo group (Hinge fracture by Takeuchi grade (0/1/2/3): Syn group 37/3/3/1, Allo group 43/8/0/0, p value=0.04*). The timing of full weight bearing in Allo group was significantly earlier than in Syn group (Weight Bearing (1=FWB, 2=PWB 3wk, 3=PWB 6wk): Syn group 2.7±0.1, Allo group 2.3±0.1, p value=0.01*). DiscussionThe use of allogenic graft to fill the void in MOWHTO does not show superiority in bone union compared to synthetic graft, however it improves pain, function, decreases the risk of hinge fracture and allows faster weight bearing than synthetic graft. Level of proofIII; Case-control study.

Synbiotic modulates intestinal microbiota metabolic pathways and inhibits DMH-induced colon tumorigenesis through c-myc and PCNA suppression

The use of probiotic and synbiotic is a promising strategy to modulate the intestinal microbiota, and thereby modify the risk of diseases. In this study, the effect of probiotic VSL#3, isolated or associated with a yacon-based product (PBY), on the functional metabolic pathways of the microbiota, in a colorectal carcinogenesis model, was evaluated. For this, mice induced to carcinogenesis were fed with standard diet AIN-93 M (CON), diet AIN-93 M and VSL#3 (PRO) or diet AIN-93 M with yacon and VSL#3 (SYN). The SYN group showed a highly differentiated intestinal community based on the MetaCyc pathways. Of the 351 predicted functional pathways, 222 differed between groups. Most of them were enriched in the SYN group, namely: amino acid biosynthesis pathways, small molecule biosynthesis pathways (cofactors, prosthetic groups, electron carriers and vitamins) carbohydrate degradation pathways and fermentation pathways. In addition, the synbiotic was able to stimulate the anti-inflammatory immune response and reduce the gene expression of PCNA and c-myc. Thus, we conclude that the synbiotic impacted more significantly the metabolic functions of the microbiota compared to the isolated use of probiotic. We believe that the enrichment of these pathways can exert antiproliferative action, reducing colorectal carcinogenesis. The prediction of the functional activity of the microbiota is a promising tool for understanding the influence of the microbiome on tumor development.

Effect of Bacillus species–fermented products and essential oils on growth performance, gut morphology, cecal short-chain fatty acid levels, and microbiota community in broilers

In this study, the effects of Bacillus species–fermented products (synbiotics [SYNs]) and essential oils (EOs) on the growth performance, gut morphology, cecal short-chain fatty acid (SCFA) levels, and microbiota of broilers were investigated. A total of 180 one-day-old unsexed broiler chicks (Ross 308) were randomly assigned to 5 dietary treatments as follows: basal diet (control group), basal diet plus enramycin (10 mg/kg; A group), basal diet plus SYNs (3 × 1011 CFU spore/kg of feed; SYN group), basal diet plus EOs (100 mg/kg; EO group), and basal diet plus SYNs and EOs (SYN + EO group), with 6 replicate cages per treatment group and 6 birds per cage. The SYN+EO treatment resulted in a higher (P = 0.003) average daily gain at 1 to 14 d of age than did the control and EO treatments. SYNs had a significant effect on the average daily gain at 1 to 14 d (P < 0.001) and 1 to 35 d (P = 0.045) of age. EOs had a significant effect on the villus height of the duodenum (P = 0.015) and jejunum (P = 0.027). Superoxide dismutase (SOD) and mucin 2 (MUC2) mRNA expression in the duodenum, jejunum, and ileum in the SYN + EO group was higher (P < 0.001) than that in any of the other groups. The SYN+EO treatment resulted in higher (P < 0.001) 2-methylbutyric acid and 3-methylbutyric acid levels in the cecal digesta of the broilers than did the control treatment. Cecal species evenness in the SYN + EO group was higher (P < 0.001) than that in the control group. The abundance of the phylum Firmicutes in the cecal digesta of the broilers was higher (P < 0.001) in the SYN+EO group than in the control group. SYNs had a significant effect (P < 0.001) on the abundance of the genus Lactobacillus in the cecal digesta of the broilers. The abundance of the genus Lactobacillus was positively associated with 2-methylbutyric acid and 3-methylbutyric acid levels. The 2-methylbutyric acid and 3-methylbutyric acid levels were positively correlated with the villus height of the duodenum and ileum. These results suggest that simultaneous supplementation with SYNs and EOs can increase the average daily gain, improve gut health–associated gene expression, increase SCFA levels, and modulate the gut microbiota composition of broilers.

Effect ofpre-synchronisation with progestogen and eCG onreproductive activity insynchronised ewes during anoestrous season.

The objective ofthe study was toinvestigate the effect ofpre-synchronisation onthe occurrence oftheoestrus and pregnancy rate after fixed time artificial insemination (FTAI) insynchronised ewes during theanoestrous season. Kivircik ewes (n=84) were randomly assigned toone ofthe two treatment groups with (PRE; n=42) orwithout (SYN; n=42) pre-synchronisation. Inthe SYN group, the ewes were subjected toa7-d short-term protocol (P4insertion-6d-PGF2α-1d-P4 removal+ eCG). In the PRE group, the same short-term protocol asintheSYN group was applied with 7 days apart for a pre-synchronised synchronisation protocol. Acervical FTAI was performed with fresh semen at54h after sponge removal. Atthe beginning ofthe synchronisation protocol, theoestrous response (66.7% vs. 0.0%) and cyclicity rates (64.3% vs. 14.3%) based onprogesterone (P4) were higher inthe PRE group compared tothose inthe SYN group, respectively (P<0.01). However, the oestrous response after synchronisation was lower within 96h (57.1% vs. 95.2%; P<0.01) inthe PRE group compared tothat intheSYN group. Although the pregnancy rate after the FTAI was significantly (P<0.05) lower inthe PRE group (14.3%) than theSYN group (35.7%), the overall pregnancy rate after natural mating was not different (95.2%) between the groups. Inconclusion, the pre-synchronisation decreased the oestrous response leading toalower pregnancy rate after theFTAI inthesynchronised ewes during the anoestrous season. Thus, attention should bepaid totwo consecutive administrations ofeCG inapre-synchronisation and synchronisation protocol inewes.

A synbiotic containing prebiotic prepared from a by-product of king oyster mushroom, Pleurotus eryngii and probiotic, Lactobacillus plantarum incorporated in diet to improve the growth performance and health status of white shrimp, Litopenaeus vannamei

This study aimed to evaluate the effects of a synbiotic composite an extract from a by-product of king oyster mushroom, Pleurotus eryngii (KOME), and probiotic Lactobacillus plantarum 7–40 on the growth performance and health status of white shrimp, Litopenaeus vannamei. The KOME was able to stimulate the growth of probiotic, but not the growth of Vibrio pathogens, including V. alginolyticus, V. parahaemolyticus, and V. harveyi. Four diets were formulated, including a control diet supplemented without prebiotic and probiotic, a basal diet supplemented with KOME (5 g kg−1) (ME), a basal diet supplemented with probiotic (1 × 108 CFU kg−1) (LP), and a basal diet supplemented with KOME (5 g kg−1) and probiotic (1 × 108 CFU kg−1) (SYN). Shrimp fed the ME, LP, and SYN diets had significantly higher survival than that of shrimp fed with the control diet for 8 weeks. Shrimp in the SYN group also had a significantly higher weight gain and total final weight in comparison with the control and other treatments. In the intestinal tract, lactic acid bacteria count was significantly higher in the SYN group, whereas the Vibrio-like bacteria count was significantly higher in the ME group than in the control group. For the health status assessment, the disease resistance of shrimp against V. alginolyticus was improved in all treatments compared to the shrimp in control. Shrimps in the SYN group had significantly lower cumulative mortality due to the significant increase in immune responses, including phenoloxidase, respiratory burst, and lysozyme activity, and the gene expression of pexn and pen4 in the haemocytes, and lgbp, sp, propoii, pexn, pen3a, pen4, and gpx in the haepatopancreas of shrimp as compared to the control. Therefore, it is suggested that a combination of KOME and probiotics can be used as a synbiotic to improve the growth performance and reduce the risk of infectious diseases caused by Vibrio and at the same time significantly contribute to the circular economy.

Syngeneic hematopoietic stem cell transplantation for acute myeloid leukemia: a propensity score-matched analysis

The present study evaluated outcomes and prognostic factors in adult patients with acute myeloid leukemia (AML) after syngeneic hematopoietic stem cell transplantation (HSCT). Among patients in first complete remission (CR1), outcomes of syngeneic HSCT (Syn) were compared with those of autologous HSCT (Auto), allogeneic HSCT from human leukocyte antigen (HLA)-matched sibling donor (MSD), or allogeneic HSCT from HLA-matched unrelated donor (MUD). Among 11,866 patients receiving first HSCT, 26 in the Syn group were analyzed. The 5-year overall survival (OS) rate, the cumulative incidence of relapse, and the cumulative incidence of non-relapse mortality (NRM) were 47.8%, 59.6%, and 4.6%, respectively. The OS was significantly better in patients in CR1 (n = 13) than in patients in non-CR1 (P = 0.012). Furthermore, 39 patients in CR1 each were assigned to the Auto, MSD, and MUD groups using propensity score matching. The 5-year OS in the Syn (68.4%) was not significantly different from those in the Auto (55.9%, P = 0.265), MSD (62.4%, P = 0.419), or MUD (63.7%, P = 0.409) groups. A higher relapse in the Syn than in the MSD and MUD groups was offset by lower NRM. In summary, syngeneic HSCT might be an alternative option for AML patients in CR1.

Effects of the Administration of Bifidobacterium longum subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001 and Their Synbiotic Combination With Galacto-Oligosaccharides Against Enterotoxigenic Escherichia coli F4 in an Early Weaned Piglet Model

We evaluated the potential of multi-strain probiotic (Bifidobacterium longum subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001) with or without galacto-oligosaccharides against enterotoxigenic Escherichia coli (ETEC) F4 infection in post-weaning pigs. Ninety-six piglets were distributed into 32 pens assigned to five treatments: one non-challenged (CTR+) and four challenged: control diet (CTR−), with probiotics (>3 × 1010 CFU/kg body weight each, PRO), prebiotic (5%, PRE), or their combination (SYN). After 1 week, animals were orally inoculated with ETEC F4. Feed intake, weight, and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized and samples from blood, digesta, and tissues collected. Microbiological counts, ETEC F4 real-time PCR (qPCR) quantification, fermentation products, serum biomarkers, ileal histomorphometry, and genotype for mucin 4 (MUC4) polymorphism were determined. Animals in the PRO group had similar enterobacteria and coliform numbers to the CTR+ group, and the ETEC F4 prevalence, the number of mitotic cells at day 4 PI, and villus height at day 8 PI were between that observed in the CTR+ and CTR− groups. The PRO group exhibited reduced pig major acute-phase protein (Pig-MAP) levels on day 4 PI. The PRE diet group presented similar reductions in ETEC F4 and Pig-MAP, but there was no effect on microbial groups. The SYN group showed reduced fecal enterobacteria and coliform counts after the adaptation week but, after the inoculation, the SYN group showed lower performance and more animals with high ETEC F4 counts at day 8 PI. SYN treatment modified the colonic fermentation differently depending on the MUC4 polymorphism. These results confirm the potential of the probiotic strains and the prebiotic to fight ETEC F4, but do not show any synergy when administered together, at least in this animal model.

Effects of Bifidobacterium longum Subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001, Combined or Not With Oligofructose-Enriched Inulin, on Weaned Pigs Orally Challenged With Salmonella Typhimurium.

Salmonella is a common causative agent of enteric disease and is developing mechanisms of resistance to antimicrobials. Probiotics, such as bifidobacteria and lactobacilli, and prebiotic fibers are a potential alternative to counteract this pathogen as they have demonstrated effectiveness in preventing its adhesion, reducing intestinal damage, and enhancing the host immune system. Furthermore, the benefits are expected to be potentiated when these compounds are administered together. A trial was performed to evaluate the efficacy of two probiotic strains (Bifidobacterium longum subsp. infantis CECT 7210 (Laboratorios Ordesa S.L.) and Lactobacillus rhamnosus HN001, combined or not with a prebiotic containing oligofructose-enriched inulin, against Salmonella Typhimurium. Ninety-six piglets (28 days old) were distributed into 32 pens assigned to 5 treatments: one non-challenged (control diet, CTR+) and four challenged: control diet (CTR−) or supplemented with probiotics (>3 × 1010 cfu/kg each strain, PRO), prebiotic (5%, PRE), or their combination (SYN). After 1 week of adaptation, animals were orally challenged with Salmonella Typhimurium. Feed intake, weight, and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized, and samples from blood, digestive content, and ileal tissues were collected to determine Salmonella counts, fermentation products, ileal histomorphology, and serum TNF-α and Pig-MAP concentrations. The effect of the oral challenge was evidenced by animal performance, fecal consistency, and intestinal architecture. Regarding the experimental treatments, animals belonging to the PRO group experienced a faster clearance of the pathogen, with more pigs being negative to its excretion at the end of the study and recovering the impaired ileal villi/crypt ratio more rapidly. Animals receiving the PRE diet showed a lower intestinal colonization by Salmonella, with no countable levels (<3 cfu/g) in any of the analyzed samples, and an augmented immune response suggested by serum Pig-MAP concentrations. Treatments including the prebiotic (PRE and SYN) showed similar changes in the fermentation pattern, with an increase in the molar percentage of valeric acid concentration in the colon. The SYN group, however, did not show any of the outcomes registered for PRO and PRE in Salmonella colonization or in immunity markers, suggesting the lack of synbiotic action in this animal model. Further research is needed to better understand the complex mechanisms behind these effects.

Treatment of locally advanced or multiple basal cell carcinoma with vismodegib in real world setting.

e22065 Background: Evaluation of efficacy and safety of vismodegib (V) was done in a retrospective analysis of patients (pts) with locally advanced or multiple basal cell carcinoma (laBCC or multiple BCC) and pts with Goltz-Gorlin Syndrom (G-G Syn) in routine clinical practice. Methods: Baseline characteristics, efficacy data and treatment-related adverse events were collected from 30 laBCC or multiple BCC and 6 G-G Syn pts who were treated with V. Results: In 86-month period, 36 pts were diagnosed with laBCC (18 pts), multiple BCC (12 pts) or G-G Syn (6 pts), all inappropriate for surgery or radiotherapy. Baseline characteristics: median age was 72.6 years in laBCC + multiple BCC pts group and 51.3 years in G-G Syn pts group. Sixty percent of pts in laBCC + multiple BCC group were females; majority (70%) of pts were previously treated by surgery (S) and/or radiotherapy (RT); 43% of pts had 1 lesion with predominant localization in central face (eyes, nose, lips or ears in 84% of pts), 20% had 2-3 lesions and 37% more than 3 lesions. Fifty percent of pts in G-G Syn pts group were males; all pts were previously treated with S and/or RT. The overall response rate (ORR) was 76% in laBCC + multiple BCC and 83% in G-G Syn pts group. Disease control rate (DCR) was 93% in laBCC + multiple BCC and 100% in G-G Syn pts group. Median duration of treatment (DoT) was 7.8 months (range: 1.3-29.8) in laBCC + multiple BCC group and 27.1 months (range: 4.8-86.4) in G-G Syn group. At the time of analysis in laBCC or multiple BCC group one patient died due to other reasons than cancer, in 30% of pts treatment has been interrupted during the treatment course [in most cases due to complete response or adverse events (AEs)], 40% of pts are still on treatment. In G-G Syn group treatment has been interrupted in 50% of pts (in most cases due to adverse events), 67% of pts are still on treatment. AEs of any grade were reported in 97% of pts in laBCC or multiple BCC group and 83% in G-G Syn group. Majority of AEs in laBCC or multiple BCC group were grade 1 or 2 (96%)., 4% of AEs were grade 3: muscle cramps in 3 pts, respiratory infection, vomiting and anemia in 1 patient each. Majority of AEs in G-G Syn group were also grade 1 or 2 (87%), 13% of AEs were grade 3: muscle cramps in 2 pts, weight loss and diarrhea in 1 patient each. No grade 4 or 5 AEs were reported. Conclusions: Vismodegib has shown meaningful efficacy with manageable safety profile in pts with laBCC or multiple BCC as well as in pts with G-G Syn in real world setting.

Formation and function of resident memory t cells in renal allografts

Abstract The role of TRM in transplantation is unknown. In this study, we investigated the formation and function of TRM in a mouse kidney transplantation model. (B6xBALB/c) F1.ova kidneys were transplanted to B6 recipients and 1m OT-I Teff cells transferred on day 2. Graft, blood, bone marrow, SLO, and liver were harvested 4 and 8 wks after transplantation. TRM were identified as CD44hiCD62LlowCD69+ CD103+/− cells after excluding in vivo labeled T cells. OT-I and polyclonal TRM were transcriptionally characterized using scRNAseq. TRM residency was tested by parabiosis and re-transplantation. To further establish a causal relationship between the TRM OT-Is and rejection we depleted them at wk 4 post adoptive transfer using anti-Thy1.1 (250 ug IV per day for 7d). Mean serum creatinine (mg/dl) was significantly higher in allo vs syn group at wk 8 (0.7 vs 0.2, p&amp;lt;0.05). Graft histology showed mixed acute and chronic rejection in the allo group. Flow analysis of allograft cells demonstrated TRM cells among OT-I and endogenous T cell populations at 4 &amp; 8 wks. The OT-I population was exclusively TRM phenotype by flow and scRNAseq, rapidly produced IFNγ upon re-stimulation, and was not detected anywhere else. There was no significant difference in mean number of OT-Is between wk 4 and wk 8 (277K vs 234k, p=0.74). OT-I T cells could not be detected in the parabiont kidney graft or tissues or in the secondary host outside the re-transplanted kidney, indicating that the TRM are indeed resident in the graft and do not re-circulate. Depleting the OT-Is preserved kidney function at wk 8 compared to the non-depleted group (Cr= 0.7 vs 0.2, p&amp;lt;0.05). Our findings show that donor-specific TRM form in kidney allografts, are functional, and contribute to rejection.

17 Combining two probiotic strains with oligofructose-enriched inulin abolishes their benefits against Salmonella Typhimurium in piglets

Abstract The aim of this study was to evaluate the efficacy of two probiotic strains Bifidobacterium longum subsp. infantis CECT7210 (Laboratorios Ordesa S.L.) and Lactobacillus rhamnosus HN001 combined or not with a prebiotic mixture of oligofructose and inulin against Salmonella typhimurium. A total of 96 piglets (28 days) were distributed into 32 pens assigned to 5 treatments: one non-challenge (control diet, CTR+) and four challenged: control diet (CTR-), or supplemented with probiotic (&amp;gt;3x1010cfu/kg each, PRO), prebiotic (5%, PRE) or their combination (SYN). After one adaptation week, animals were inoculated with Salmonella. Feed intake, weight and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized and samples collected for Salmonella counts, fermentation products, ileal histomorphology and serum inflammatory markers. After the challenge, feed intake was decreased (P = 0.001) more markedly in the SYN group with a lower final weight (P = 0.009). PRE and SYN groups showed more liquid ileal consistency on day 8PI (P = 0.009). A higher percentage of animals receiving PRO became negative to Salmonella in feces at the end of the study (65% PRO vs 0% CTR-, P = 0.028). Animal receiving PRE did not reach countable levels in feces any time during the trial (vs 23.2% for CTR-group, P = 0.013). Histologically, on day 4PI a decrease in the ileal villous/crypts ratio was found in challenged animals (P &amp;lt; 0.001), being PRO able to improve the recovery from this damage at day 8PI (P = 0.008). Animals receiving PRE had higher number of intraepithelial lymphocytes (IEL) on day 8PI (P = 0.028). In conclusion, the probiotic strains were able to promote a faster clearance of the pathogen from the gut with a faster recovery of damaged histological architecture. The prebiotic reduced Salmonella colonization and stimulated the local immune response. However, these beneficial effects disappeared when they were administered in a synbiotic form.

Effect of synbiotic dietary supplementation on histological and histopathological parameters of Pectoralis major muscle of broiler chickens

Bioactive substances may constitute an alternative to antibiotics. Probiotics are mixtures of microorganisms that enhance the effectiveness and yield of nutrient absorption by competing for the substrate against pathogens that may cause intestinal infections. Prebiotics are organic substances which include complex carbohydrates and have an influence on the growth and activity of desirable intestinal microflora. Synbiotics are a combination of both of them. The aim of this study was to analyze the impact of synbiotics added to commercial feed on Pectoralis major muscle microstructure in broiler chickens. Research material consisted of 240 one-day-old Ross 308 female chicks. Birds were divided into 2 groups, 12 broilers each (replicated 10 times). The control group (C) was fed a commercial diet, and the SYN group was fed the same diet with added synbiotic: 0.8% of prebiotic RFO (raffinose family oligosaccharides) extracted from lupin seeds and 1% of probiotic Lavipan&lt;sup&gt;®&lt;/sup&gt;. Birds were slaughtered at day 42. Immediately after slaughter, samples of the Pectoralis major muscle for histological analysis were taken and frozen in liquid nitrogen. The staining procedures performed were: hematoxylin and eosin staining to evaluate fibre diameter and histopathological changes, oil red staining to determine intramuscular fat content, NADH-TR (tetrazolium reductase) activity to distinguish muscle fibre types and alkaline phosphatase reaction for counting the number of capillaries. The tendency to reduced muscle fibre diameter in chickens supplemented with synbiotics indicates an increase in muscle fibre density, which gives meat a more delicate structure. When assessing the occurrence of histopathological changes, significantly more fibre splitting was observed in the control group. Moreover, the positive effect of feed supplementation with synbiotics on muscle microvascularization could have an important practical meaning, because low physical activity of chickens during rearing may lead to ischemic muscles, increased changes in the structure of muscle fibres, and reduction of meat quality.

A synbiotic improves the immunity of white shrimp, Litopenaeus vannamei: Metabolomic analysis reveal compelling evidence

In this study, we examined the synergistic effects of a diet-administered synbiotic comprising galactooligosaccharide (GOS) and the probiotic Lactobacillus plantarum 7–40 on immune responses, immune-related gene expressions, and disease resistance to Vibrio alginolyticus in white shrimp Litopenaeus vannamei. To unravel the regulatory role of the synbiotic in activating the immune system of shrimp, 1H nuclear magnetic resonance (NMR)-based metabolomic analysis were used to investigate hepatopancreas metabolites, then significantly altered metabolites were confirmed in both the hepatopancreas and plasma by reverse-phase high-performance liquid chromatography (RP-HPLC) and spectrophotometric analysis. Shrimp were fed four experimental diets for 60 days, including a basal diet with no GOS or probiotic (control), 0.4% GOS (PRE), probiotic (PRO), and 0.4% GOS in combination with the probiotic (SYN). Results showed that the SYN diet significantly increased survival of L. vannamei 24 h after a V. alginolyticus injection. Immune parameters such as phenoloxidase activity, respiratory bursts, phagocytic activity and gene expressions, including prophenoloxidase I, serine proteinase, and peroxinectin, of shrimp fed the SYN diet significantly increased, compared to the other treatments and control. In addition, results from the 1H NMR analysis revealed that 22 hepatopancreas metabolites were matched and identified between the SYN and control groups, among which three metabolites, i.e., inosine monophosphate (IMP), valine, and betaine, significantly increased in the SYN group. Confirmation using RP-HPLC and spectrophotometric methods showed that IMP presented high amounts in the hepatopancreas, but not in the plasma of shrimp; in contrast, valine and betaine metabolites were in high concentrations in both the hepatopancreas and plasma. Our results suggested that GOS and the probiotic had a synergistic effect on enhancing immunity and disease resistance of L. vannamei against V. alginolyticus infection through inducing syntheses of a nucleotide (IMP), a branched amino acid (valine), and a methyl group donor (betaine) in the hepatopancreas, which were then released into the plasma and directly taken up by hemocytes, resulting in a triggering of melanization and phagocytosis processes in cells.

Effects of mannan-oligosaccharides and Lactobacillus acidophilus supplementation on growth performance, nutrient utilization and faecal characteristics in Murrah buffalo calves.

A study of 120days was undertaken to ascertain the effect of mannan-oligosaccharides (MOS) and Lactobacillusacidophilus supplementation on growth performance, nutrient utilization and faecal characteristics in Murrah buffalo calves. Twenty Murrah buffalo calves of 5-7 days old and 31±2.0kg of body weight (BW) were randomly assigned into four groups. Group I served as the control (CON) in which only basal diet (concentrate mixture and green fodder) was provided, without any supplementation. Mannan-oligosaccharides at 4g/calf/day were supplemented as prebiotic to Group II (PRE), whereas Group III (PRO) received Lactobacillusacidophilus in the form of fermented milk as probiotic at 200ml/calf/day having 108 CFU/ml and Group IV (SYN) was supplemented with both MOS and Lactobacillusacidophilus as synbiotic at similar dose. Final BW (kg), dry matter intake, average daily gain, feed conversion efficiency and structural growth measurements were improved (p<.05) in the treatment groups compared to control. Digestibility of neutral detergent fibre was higher (p<.05) in SYN followed by PRE and PRO than control. The faecal lactobacilli and bifidobacterium population was higher (p<.05) in all the supplemented groups with a concomitant reduction in faecal coliform count as compared to control. Faecal ammonia, lactate and pH were also altered favourably (p<.05) in all the supplemented groups as compared to CON. The faecal volatile fatty acids were higher (p<.05) in PRE, PRO and SYN group than CON. The incorporation of MOS and Lactobacillusacidophilus in diet either individually or in combination as synbiotic has the potential to improve the performance and faecal characteristics in Murrah buffalo calves; however, the observed responses among the treatment groups were more evident in the synbiotic fed group compared to individual supplementation of MOS and Lactobacillusacidophilus.

Effects of dietary supplementation with Pediococcus acidilactici MA18/5M, galactooligosaccharide and their synbiotic on growth, innate immunity and disease resistance of rockfish (Sebastes schlegeli)

The trial investigated the effects of dietary supplementation with the commercial prebiotic galactooligosaccharide (1% GOS; GOS diet), the commercial probiotic Pediococcus acidilactici MA18/5M (6.3 log CFU g−1 PA; PA diet) and their combination as synbiotic (1% GOS and 6.3 log CFU g−1 PA; SYN diet) on the growth, innate immunity and disease resistance of juvenile rockfish (Sebastes schlegeli). Triplicate groups of fish (18.2±0.3g) were fed one of the four diets for 8weeks and then subjected to a standard challenge with Edwardsiella tarda by intraperitoneal injection. The results showed significant (P<0.05) improvement in growth performance in the SYN compared to the control group with intermediary benefits for PA and GOS. White blood cells count was significantly enhanced in the PA and GOS groups with a further significant increase for the SYN diet. In accordance and compared to the control; serum lysozyme and bacteriostatic activities against Vibrio anguillarum and Escherichia coli were increased in all supplemented groups while serum alternative complement activity was elevated in the SYN group. In addition, the skin mucus bacteriostatic activities were significantly influenced by dietary treatments. The beneficial effects observed on circulating innate immune parameters were validated by a higher survival pattern following intraperitoneal challenge with E. tarda in the PA and SYN groups. This study contributes to the knowledge on the mode of action of GOS and PA and demonstrates their potential as natural health and growth promoters in the rockfish on-growing aquaculture industry.

Effects of microencapsulated Lactobacillus plantarum and fructooligosaccharide on growth performance, blood immune parameters, and intestinal morphology in weaned piglets

ABSTRACT This experiment aimed to investigate the effects of a symbiotic, consisting of microencapsulated Lactobacillus plantarum (MLP) and fructooligosaccharide (FOS) on growth performance, blood immune parameters, and intestinal microbiota in weaned piglets. Ninety weaned piglets were assigned to three dietary groups (CON: basal diet; ANT: basal diet + aureomycin; SYN: basal diet + MLP and FOS) for a four-week trial. Compared to CON, pigs in the SYN group had higher weight gain and feed intake, and lower diarrhea rate (P < .05). Also, pigs in the SYN group had improved plasma IgA and IgG concentrations, and increased (P < .05) numbers of lactic acid bacteria in the colon compared to CON. In conclusion, feeding a synbiotic based on MLP and FOS had beneficial effects on growth performance, plasma immune parameters, and intestinal microbiota, indicating the potential of it to serve as an alternative to feed antibiotics in weaned pig diets.

Impacts of albumin synergized with hydroxyethyl starch on early microvascular albumin leakage after major abdominal surgery in rabbits.

To investigate the impacts of albumin synergized with hydroxyethyl starch (HES) on early microvascular albumin leakage after major abdominal surgery in rabbits. Forty male Japanese rabbits were randomly divided into four groups: the control group, the saline group, the albumin group, and the Syn group (hydroxyethyl starch+albumin). The latter three groups were performed gastrectomy plus resection of pancreatic body and tail and splenectomy. The serum albumin concentration was detected before and 48h after surgery, and the conditions of mesenteric microvascular leakage in these 4 groups were observed under microscope 48 h after surgery to calculate the leakage rate. Compared with the saline group, the albumin group and the Syn group exhibited significantly increased serum albumin concentrations 48h after surgery (P<0.05). The albumin leakage rate was the most obvious in the albumin group, followed by the saline group, while that in the Syn group was the minimal, and there existed significant differences among these groups (P<0.05) . Simple administration of albumin in the early stage after major abdominal surgery could increase the albumin leakage, while the synergization of albumin and hydroxyethyl starch could reduce the albumin leakage.