D-lactate Levels Research Articles

Comparison of Deep and Moderate Neuromuscular Blockade on Intestinal Mucosal Barrier in Laparoscopic Gastrectomy: A Prospective, Randomized, Double-Blind Clinical Trial.

Deep neuromuscular blockade (NMB) improves the surgical conditions and is benefit for the postoperative recovery after laparoscopic surgery. However, the mechanisms of deep NMB in promoting the recovery of intestinal function have not been completely investigated. The objective of our study was to determine the impact of the deep NMB and moderate NMB strategy on the intestinal barrier function after laparoscopic gastrectomy. We collected patients undergoing elective laparoscopic gastrectomy. Patients were randomized to deep NMB (post-tetanic count 1–2) vs. moderate NMB (train-of-four count 1–2) during the surgery. Primary outcomes were time to flatus, serum diamine oxidase (DAO) and D-lactate, and gut microbiota. Other outcomes were surgical condition scores, postoperative visual analog pain scores, and length of hospital stay. Ninety patients in deep NMB group and sixty patients in moderate NMB group completed the study. Main results showed that the time to flatus was decreased in deep NMB group (74 ± 32 h) than that in moderate NMB group (93 ± 52 h, P = 0.006). The level of serum D-lactate was statistically increased in the moderate NMB group than that in the deep NMB group (1,209 ± 224 vs. 1,164 ± 185 ng/ml, p < 0.001). But no significant differences could be detected in the level of DAO between the groups. Additionally, the 16s rRNA analysis indicated that gut microbiota were similar in Alpha diversity but distinct in Beta diversity. Furthermore, the beneficial bacteria, such as genus Lactobacillus and Bifidobacterium, were more abundant in the deep NMB group, while the potentially harmful bacteria were more abundant in the moderate NMB group. Our findings suggested that the intestinal mucosal barrier and gut microbiota were better preserved in deep NMB, which greatly improved the postoperative recovery of intestinal function after laparoscopic gastrectomy.

The Gut-Liver Axis in Nonalcoholic Fatty Liver Disease: Association of Intestinal Permeability with Disease Severity and Treatment Outcomes.

Objective To investigate the association between intestinal permeability and severity of nonalcoholic fatty liver disease (NAFLD) and the value of intestinal permeability in predicting the efficacy of metabolic therapy for NAFLD. Methods Disease severity was compared between patients with normal and elevated intestinal permeability; correlations between D-lactate and different NAFLD parameters were analyzed; and the effects of metabolic therapy on NAFLD patients with normal and elevated intestinal permeability were evaluated. Results A total of 190 patients with NAFLD were enrolled. NAFLD patients with elevated intestinal permeability had significantly higher levels of liver test parameters, liver ultrasonographic fat attenuation parameter, triglyceride, homeostasis model assessment of insulin resistance value, and diamine oxidase (all P˂0.05) than NAFLD patients with normal intestinal permeability. Furthermore, serum D-lactate levels were positively correlated with alanine transaminase, aspartate transaminase, gamma-glutamyl transpeptidase, total bilirubin, indirect bilirubin, fat attenuation parameter, triglyceride, and diamine oxidase (all P ˂ 0.05). Moreover, NAFLD patients with elevated intestinal permeability showed less improvement in TG levels (P = 0.014) after metabolic therapy. Conclusion Intestinal permeability correlates with the disease severity in patients with NAFLD. Moreover, intestinal permeability may have value for predicting the efficacy of metabolic therapy for NAFLD patients.

Integrin αEβ7 is involved in the intestinal barrier injury of sepsis.

Background: IL-9-producing CD4(+) T (Th9) cell was related to acute intestinal barrier injury in sepsis. Integrin αEβ7 was an important lymphocyte homing receptor on the surface of intestinal Th9 cells. However, the roles of αEβ7 in the intestinal injury caused by Th9 cells were not clear in sepsis.Methods: To investigate the roles of αEβ7 in the intestinal injury caused by Th9 cells in sepsis model, the Th9 cells percentages, αEβ7, E-cadherin, IL-9, and D-lactate levels in both serum and intestinal tissue were measured. The intestinal histopathology, epithelium apoptosis, and mucosal permeability measurement were also performed. The survival rate of septic rats was recorded daily for 14 days.Results: Rats were assigned to four cohorts: control cohort, sepsis cohort, sepsis+αEβ7i (αEβ7 inhibition) cohort, and sepsis+αEβ7e (αEβ7 overexpression) cohort. The Th9 cells percentages, αEβ7, IL-9, and D-lactate levels of the sepsis cohort were significantly higher than those of the control cohort. The levels of these variables were also elevated progressively in the sepsis+αEβ7i cohort, sepsis cohort, and sepsis+αEβ7e cohort. The E-cadherin levels were decreased progressively in the control cohort, sepsis+αEβ7i cohort, sepsis cohort, and sepsis+αEβ7e cohort. Moreover, αEβ7 overexpression could decrease the 14-day survival rate. The findings of histopathology staining, apoptosis detection, and intestinal permeability test also confirmed that the barrier injury was deteriorated or relieved by elevating or decreasing the αEβ7 expression levels, respectively.Conclusions: Integrin αEβ7 was closely associated with the intestinal barrier injury caused by Th9 lymphocytes in sepsis.

Intestinal Microbiota and Serum Metabolic Profile Responded to Two Nutritional Different Diets in Mice.

There is an interaction and bidirectional selection between dietary intake and gut microbiota due to the different efficiency of nutrients in the gut. The nutritional composition of germ-free (GF) diets differs significantly from specific pathogen-free (SPF) diets. There is, however, no data revealing how SPF animals from the same microbial background respond to them and if they affect the host. We examined the growth of SPF mice on the GF diet and found that it reduced body weight, intestinal length and intestinal morphology. Interestingly, the GF diet increased the level of pro-inflammatory bacteria in the gut of SPF mice, including Proteobacteria, Burkholderiaceae, Alloprevotella and Parasutterella. Furthermore, GF diets caused significant increases in malondialdehyde (MDA), IL-1β, IL-6, and D-lactate levels in the serum of SPF mice and significantly altered their serum metabolic profile, especially amino acid metabolism. In conclusion, GF diets are not suitable for the growth and development of SPF mice. These findings, based on the role of gut microbiota in diet selection, provide new insights into the scientific and rational use of experimental animal diets.

Protective effects of nanocurcumin against stress-induced deterioration in the intestine

The therapeutic activities of curcumin have long been investigated in some chronic and inflammatory diseases. This study was designed to investigate the protective effects of nanocurcumin on intestinal barrier function, apoptosis, and oxidative stress in rats exposed to traffic noise. Forty rats were divided into four groups: two traffic noise-exposed groups of animals that received either vehicle (NOISE) or nanocurcumin (NCUR + NOISE) and two control groups that either remained intact (CON) or received nanocurcumin (NCUR). Nanocurcumin injection (15 mg/Kg/ip) and traffic noise exposure were administered daily for two weeks. The relative protein expression of intestinal tight junctions, occludin, and ZO-1 and Bax/Bcl-2 ratio was measured to evaluate barrier integrity and apoptosis in intestinal samples, respectively. Plasma D-lactate concentration was examined as a criterion of intestinal permeability. Corticosterone, superoxide dismutase (SOD) activity, glutathione (GSH), total antioxidant capacity (TAC), and nitrite were measured in serum. The noise exposure increased Bax/Bcl-2 ratio, corticosterone, and oxidative stress in the NOISE animals. Nanocurcumin treatment improved the Bax/Bcl-2 ratio and reduced corticosterone and oxidative stress in the NCUR + NOISE animals. The expression of tight junction proteins was decreased while the concentration of D-lactate was increased in the NOISE animals. Nanocurcumin did not efficiently impact the expression of tight junction proteins and the D-lactate level in the NCUR + NOISE group. Nanocurcumin administration displayed antioxidant and anti-apoptotic roles in the noise-exposed rats, however, it did not affect the intestinal barrier integrity. We concluded that reduced apoptosis in the intestine might be related to the antioxidant activity of nanocurcumin and its modulatory effects on the HPA axis in the nanocurcumin-treated animals.

Clostridium butyricum powders combined with zinc gluconate on inflammatory factors and intestinal microecology in children with infectious diarrhea

Objective: To investigate the clinical value of Clostridium butyricum powders combined with zinc gluconate in the treatment of infectious diarrhea in children, clinical data of 86 children with infectious diarrhea were retrospectively analyzed. Methods: Both groups received routine treatments to reduce symptoms, among which the control group ( n = 40) was given Clostridium butyricum powders, and the experiment group ( n = 46) was given Clostridium butyricum powders and zinc gluconate. Patients in both groups were treated for 5 days. The clinical efficacy, recovery time, safety, serum levels of inflammatory markers (interleukin-6, interleukin-17, C-reactive protein), indicators of the intestinal mucosal function (diamine oxidase, D-lactate) and indicators to intestinal microecology (bifidobacteria, lactobacillus) before and after treatment were compared between the two groups. Results: The clinical efficacy in the experiment group (95.65%) was higher than that in the control group (77.50%, p &lt; 0.05). The time of recovery from symptoms including diarrhea, abdominal pain and vomiting, and to normal body temperature in the experiment group were shorter than those in the control group (all p &lt; .001). After treatment, the serum IL-6, IL-17, CRP levels and DAO, D-lactate levels in both groups were lower than those before treatment, and were lower in the experiment group than in the control group (all p &lt; .001). After treatment, the intestinal bifidobacteria and lactobacillus levels in both groups were higher than those before treatment, and were higher in the experiment group than in the control group (all p &lt; .01). There was no significant difference in the incidence of adverse reactions between the experiment group (8.70%) and the control group (2.50%). Conclusion: In this open-label study, Clostridium butyricum powders combined with zinc gluconate are significantly effective in treating children with infectious diarrhea, which is conducive to relieving early symptoms, downregulating serum inflammatory factor levels, improving intestinal mucosal function, regulating intestinal microecology, and ensuring the safety of patients.

An unusual cause of acute encephalopathy: D-lactic acidosis secondary to short bowel syndrome

Background: Blood D-lactate levels increase in short bowel syndrome (SBS) and may lead to neurological manifestations. Clinical Description: A 5-year-old boy, postoperative case of SBS, presented with loose stools, generalized weakness, and lethargy for 2 days. The child had undergone significant intestinal resection in the past. On examination, he had some dehydration, and was drowsy, but arousable. Remaining examination was normal. Metabolic abnormalities detected included metabolic acidosis (pH of 7.1, HCO3 7 mmol/L), high anion gap (20 mmol/l), and normal lactate levels (2 mmol/L). Other baseline investigations were normal. He was treated as a case of acute gastroenteritis with some dehydration and metabolic acidosis and improved. He was discharged after 5 days. After 2 months, he was readmitted with drowsiness and unsteady gait. This time there was no diarrhea or dehydration. Investigations again revealed severe metabolic acidosis, high anion gap, and normal lactate levels. Management: We considered SBS induced D-lactate encephalopathy but were unable to prove it by assay due to unavailability of tests. The child was kept nil per orally and given bicarbonate infusion, on which he showed dramatic improvement. He was also given a low carbohydrate diet and oral metronidazole. The family was counseled at discharge 5 days later regarding dietary modifications and microsupplementation. The patient had 6 admissions for D-lactic encephalopathy over 4 years that coincided with dietary lapse. Conclusion: D-lactate acidosis is an underrecognized condition and its diagnosis and management is challenging. A high index of suspicion should be kept in patients with history of intestinal resection presenting with acute encephalopathy and unexplained metabolic acidosis.

Elevated Levels of Circulating Biomarkers Related to Leaky Gut Syndrome and Bacterial Translocation Are Associated With Graves' Disease.

BackgroundA growing number of studies have found dysbiosis of the intestinal microbiota in patients with Graves’ disease (GD). The intestinal epithelial barrier serves as the first line of defense, protecting the immune system from excessive stimulation of microbiota and toxins. Most autoimmune diseases are associated with a gut barrier dysfunction (leaky gut) which allows bacterial translocation. However, to date, potential correlations between intestinal barrier dysfunction and GD have not been explored.MethodsSerum lipopolysaccharide (LPS), intestinal fatty acid-binding protein (I-FABP), zonulin, D-lactate, and diamine oxidase (DAO) were measured to assess barrier integrity in 91 patients with GD (61 initial GD and 30 euthyroid GD) and 44 healthy controls. The quality of life (QOL) of patients with GD was assessed using the thyroid-specific patient-reported outcome (ThyPRO-39) questionnaire.ResultsThe serum levels of LPS, I-FABP, zonulin, and D-lactate were significantly higher in patients with initial GD than in healthy controls. Logistic regression analysis revealed that zonulin and D-lactate were independently associated with risk for GD and circulating zonulin could effectively distinguish patients with initial GD from healthy controls. Correlation analyses showed that I-FABP, LPS, and D-lactate were positively associated with FT4 and negatively associated with TSH. In addition, circulating LPS, zonulin, and D-lactate levels were all independent predictors of TRAb levels. Moreover, higher circulating LPS levels in patients with GD were associated with more severe hyperthyroidism (higher concentrations of FT3, FT4, and TRAb and lower TSH concentrations) and worse scores of hyperthyroid and eye symptoms.ConclusionPatients with initial GD show a disrupted intestinal barrier, characterized by elevated levels of leaky gut biomarkers. Increased intestinal permeability and bacterial translocation were associated with TRAb levels and hyperthyroidism in GD. Further research is required to elucidate the underlying mechanisms.

Effects of L-theanine on intestinal morphology, barrier function, and MAPK signaling pathways in diquat-challenged piglets

This study aimed to explore the protective effects of L-theanine supplementation on the diquat-challenged weaned piglets. A total of 160 weaned piglets were randomly divided into 4 groups using a 2 × 2 two-factor design, there were 4 replicates per group and 10 pigs per replicate. Piglets were fed diets (with 1000 mg/kg L-theanine addition or not), then challenged with diquat or saline on day 7. 21 days after challenge, two pigs from each replicate were selected for sample collection. Results showed that supplement with 1000 mg/kg L-theanine down-regulated the diarrhea rate, serum D-lactate level, tumor necrosis factor-α, and phosphorylation of extracellular regulated protein kinases (ERK1/2), p38 mitogen-activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK) signaling in pigs without diquat challenge (p < 0.05). While for diquat-challenged piglets, L-theanine addition increased average daily gain, jejunum villus height, and interferon-γ level (p < 0.05). Meanwhile, L-theanine addition decreased the diarrhea rates and mortality, serum D-lactate level, and phosphorylation of ERK and JNK in diquat-challenged pigs (p < 0.05). These results demonstrate that L-theanine pretreatment could alleviate diquat-induced oxidative stress and improve intestinal barrier function in diquat-challenged weaned piglets, which can be attributed to suppression of MAPK phosphorylation signaling pathways.

Dietary Seleno-l-methionine Alters the Microbial Communities and Causes Damage in the Gastrointestinal Tract of Japanese Medaka Oryzias latipes

Excess dietary seleno-l-methionine (Se-Met) induces various adverse effects in fish inhabiting the Se-contaminated environments. However, there is an extreme paucity of data on the effects of excess dietary Se-Met on the microbiota in the gastrointestinal (GI) tract in fish. In this study, Japanese medaka Oryzias latipes (three months old) were fed the Se-Met enriched diets at environmentally relevant concentrations: 2.90 (Control: (C), 6.69 (L), 11.89 (M), and 27.05 (H) μg Se/g dw) for 60 d. Histopathological, high throughput sequencing, and biochemical approaches were used to investigate the alterations in histology and microbial communities of the GI tract, enzymatic activity, and transcripts of closely related genes. The results showed that the fish weight was reduced at ∼13% from the L and H treatments. Decreased height and thickness of villus in the GI tract were observed in the H treatment. Meanwhile, the level of D-lactate and activity of diamine oxidase (DAO), protease, and lipase were inhibited in the H treatment. The transcripts of the genes related to the inflammation (i.e., IL-1β and IL-8) were elevated, while those of the genes related to the intestinal barrier (i.e., cdh1, ZO-1, ocln, and cldn7) were inhibited in the H treatment. In addition, alpha diversity at the genus level was higher in the L treatment than the control, and the composition of the microbial community was altered by dietary Se-Met. Furthermore, 5 genera (Rhodobacter, Cloacibacterium, Bdellovibrio, Shinella, and Aeromonas) exhibited the largest variation in abundance among treatments. This study has demonstrated that excess dietary Se-Met inhibits growth, causes hispathological damage to the GI tract, and alters the composition of the microbial community in Oryzias latipes.

Dietary palygorskite-based antibacterial agent supplementation as an alternative to antibiotic improves growth performance, intestinal mucosal barrier function, and immunity in broiler chickens

The aim of this study was to evaluate effects of palygorskite-based antibacterial agent (PAA) as an alternative to antibiotic on growth performance, intestinal barrier function, and immunity in broilers. Three hundred and eighty-four mixed-sex 1-day-old Ross 308 broiler chicks were allocated into 6 groups of 8 replicates with 8 birds each. Birds were given a basal diet, an antibiotic diet (50 mg/kg chlortetracycline), and the basal diet supplemented with 250, 500, 1,000, and 2,000 mg/kg PAA for 42 d, respectively. Compared with control group, supplementing 1,000 mg/kg PAA reduced overall feed conversion ratio (P < 0.05), with its value being similar to that of antibiotic group (P > 0.05). However, a higher level of PAA (2,000 mg/kg) increased feed conversion ratio during the late period (P < 0.05). The 1,000 and 2,000 mg/kg PAA decreased plasma endotoxin and D-lactate levels at 42 d (P < 0.05) to comparable values (P > 0.05). The 1,000 mg/kg PAA decreased jejunal crypt depth, while 500 and 1,000 mg/kg PAA increased the ratio between jejunal villus height and crypt depth at 42 d (P < 0.05), with their values being similar to antibiotic group (P > 0.05). The highest level of PAA increased 42-d jejunal mucosal secretory immunoglobulin A and immunoglobulin M concentrations (P < 0.05). The 1,000 and 2,000 mg/kg PAA reduced 21-d interleukin-1β and tumor necrosis factor-α (TNF-α) levels in serum and ileal mucosa and 42-d interferon-γ level in serum and jejunal mucosa (P < 0.05), which did not differ from antibiotic group (P > 0.05). Moreover, PAA administration, regardless of its dosage, reduced 42-d serum TNF-α concentration, and 500 to 2,000 mg/kg PAA decreased 21-d and 42-d jejunal and 42-d ileal mucosal TNF-α levels (P < 0.05), with their values being comparable with antibiotic group (P > 0.05). The results suggested that PAA as an alternative to antibiotic could improve growth performance, intestinal barrier function, and immunity of broilers, and its optimal dosage was 1,000 mg/kg.

CCAAT/Enhancer-Binding Protein Homologous Protein (CHOP) Deficiency Attenuates Heatstroke-Induced Intestinal Injury

The intestine is one of the main target organs involved in the pathological process of heatstroke. CCAAT/enhancer-binding protein homologous protein (CHOP) is involved in endoplasmic reticulum (ER) stress-induced apoptosis. This study aimed to explore the role of CHOP in heatstroke-induced intestinal injury and potential therapy. An in vitro heat stress (HS) model using Caco-2 cells was employed. We observed the role of CHOP in apoptosis-mediated intestinal epithelial cell injury secondary to HS by evaluating cell viability, lactate dehydrogenase release, apoptosis levels, and GRP78, PERK, ATF4, CHOP, Bcl-2, and BAX mRNA and protein expression. To further study the role of CHOP in HS-induced intestinal barrier dysfunction, we assessed transepithelial electrical resistance, paracellular tracer flux, ultrastructure of tight junctions, and protein expression of ZO-1 and occludin. Male wild-type mice and CHOP knockout mice were used for in vivo experiments. We evaluated serum d-lactate and diamine oxidase levels, histopathological changes, intestinal ultrastructure, and ZO-1 and occludin protein expression. HS activated the PERK-CHOP pathway and promoted apoptosis by upregulating BAX and downregulating Bcl-2; these effects were prevented by CHOP silencing. Intestinal epithelial barrier function was disrupted by HS in vitro and in vivo. CHOP silencing prevented intestinal barrier dysfunction in Caco-2 cells, whereas CHOP knockout mice exhibited decreased intestinal mucosal injury. The ER stress inhibitor 4-phenylbutyrate (4-PBA) prevented HS-induced intestinal injury in vitro and in vivo. This study indicated that CHOP deficiency attenuates heatstroke-induced intestinal injury and may contribute to the identification of a novel therapy against heatstroke associated with the ER stress pathway.

Analysis of pH and electrolytes in blood and ruminal fluid, including kidney function tests, in sheep undergoing long-term surgical procedures

BackgroundThe physiology of sheep as small ruminants is remarkably different from monogastric animals especially regarding the forestomach system. Using sheep for surgical procedures during scientific research thereby presents an exceptional setting for the anaesthetist. Long-term anaesthesia generally demands deprivation of food to reduce the risk of bloat in sheep. This might influence the energy and electrolyte balance. In horses and companion animals, close monitoring of mean arterial blood pressure, capnography and blood gas analysis are common procedures during long-term surgery. However, few data are available on reference ranges for blood gas in sheep and these cover only short periods of anaesthesia. To the authors’ knowledge, there is no study available that includes the monitoring of electrolytes and pH in ruminal fluid and kidney function tests in sheep undergoing long term anaesthesia. Thereby, the aim of the present study was to gather data on blood parameters, and data on ruminal fluid and kidney function during long-term anaesthesia in sheep. Data were obtained from eight sheep undergoing the invasive surgical procedure of left pneumonectomy and auto-transplantation or isolated left lung perfusion. After a 19-h fasting period, the animals were administered xylazine and ketamine and then intubated and maintained in general anaesthesia under artificial ventilation using isoflurane in oxygen. Blood samples were evaluated during 9 h of anaesthesia; ruminal fluid and kidney function tests were evaluated during 7 h of anaesthesia.ResultsBlood parameters such as electrolytes and partial pressure of carbon dioxide revealed few changes, yet blood glucose decreased and beta-hydroxybutyric acid increased significantly. All animals showed an elevated arterial pH and bicarbonate concentration despite artificial ventilation. In ruminal fluid, the pH significantly decreased and no significant changes in electrolytes occurred. Kidney function tests revealed no significant changes in any of the animals. However, fractional excretion of water and phosphate was slightly increased. One animal showed severe complications due to hypokalaemia.ConclusionInvasive surgery under long-term anaesthesia in sheep is possible without great imbalances of arterial pH and electrolytes. Nevertheless, potassium concentrations should be monitored carefully, as a deficiency can lead to life-threatening complications. The operated sheep tended not to develop metabolic acidosis and the mean kidney function could be maintained within the physiological range throughout anaesthesia. However, slight elevations in renal fractional water and phosphate excretion could suggest an early tubular reabsorption dysfunction. In ruminal fluid, acidification occurred, though no significant changes were observed in l- and d-lactate levels or in electrolyte concentrations. To our knowledge, the role of the rumen in storing fluids and balancing electrolytes in the blood has not yet been documented during anaesthesia. However, the importance of the rumen for fluid equilibrium in sheep indicates the necessity for routine monitoring and further research.

ApoB48 (remnant) Dyslipoproteinemia Correlates with D-lactate, an Advanced Glycation Precursor Catabolic Product, in Obese Adolescents

BackgroundMethyglyoxal (MG) is a toxic molecule that mediates damage produced by glycation, a key pathogenic factor in diabetes mellitus. We have previously shown that the MG catabolic product, D-lactate, is higher in obese adolescents (1) and that apoB48 is higher in adolescents with insulin resistance (IR) (2). Hypothesis and aims: We tested the hypothesis that increased D-lactate is associated with ApoB48 (remnant) dyslipoproteinemia and IR in adolescents. Material and MethodsThis cross-sectional study in anew cohort included euglycemic adolescents between 15 and 19 years old, classified in 4 groups according to BMI and HOMA-IR: lean adolescents without (MHL) alterations (n = 20), lean adolescents with metabolic alterations (MUL, n=20) metabolically healthy obese (MHO) n = 20 and metabolically unhealthy obese adolescents (MUO) n = 24. ResultsD-lactate levels did not differ between MHL and MUL participants (0.29 +/- 0.15 vs 0.29 +/- 0.13 mmol/L), they show a trend to be higher in MHO (0.38 +/- 0.15 mmol/L) and are significantly higher (0.46 +/- 0.15 mmol/L) in MUO vs MUL or MHL (p=0.003. D-lactate correlated with BMI (Rho=0.39, p=0.0002); HOMA-IR (Rho=0.33, p=0.002) and apoB48 HOMA-IR (Rho=0.33, p=0.002). Conclusion: MUO displayed 58% higher levels of D-lactate than lean adolescents. D-lactate correlates with BMI, HOMA-IR and with chylomicron remnants. We suggest that IR does not suffice to produce increased MG metabolism (MHL and MUL have the same levels) but potentiates the effect of obesity. Increased D-lactate show dysregulation of glycolysis leading to MG production, in association with poor chylomicron catabolism. 1. Rodríguez-Mortera, Reyna, Claudia Luevano-Contreras, Sergio Solorio-Meza, Russell Caccavello, Yasmin Bains, Ma Eugenia Garay-Sevilla, and Alejandro Gugliucci. Clinical Chemistry and Laboratory Medicine (CCLM) 56, no. 7 (2018): 1100-1108. 2. Rodríguez-Mortera R, Caccavello R, Garay-Sevilla ME, Gugliucci A. Clin Chim Acta. 2020; 508:61-68. doi:10.1016/j.cca.2020.05.014

Design and development of novel fasudil derivatives as potent antibreast cancer agent that improves intestinal flora and intestinal barrier function in rats.

This study was conducted to develop novel fasudil derivatives after incorporation of substituted thiazoles as potent anti-breast cancer (BC) agents. The compounds were developed using a facile synthetic route in excellent yields. The entire set of developed compounds was tested for inhibitory activity against rho-associated coiled-coil kinase (ROCK; ROCK1 and ROCK2) kinase, where they exhibit potent and selective inhibition of ROCK1 as compared to ROCK2. The most potent ROCK2 inhibitor, compound 6h significantly inhibited the viability of BC cells (MCF-7). It also causes inhibition of migration and invasion of MCF-7 cells. Moreover, the anti-BC activity of compound 6h was studied in 7,12 dimethyl Benz(a)anthracene (DMBA)-induced BC in female Sprague Dawley rats. Results suggest that it causes significant improvement in the bodyweight of the animals with a reduction in oxidative stress in the liver and mammary tissues of rats. It showed improvement in the intestinal barrier function of rats by restoring the level of Diamine oxidase, d-lactate, and endotoxin. In western blot analysis, it showed improvement in (ZO-1), occludin, and claudin-1 in the colon tissue of the rat as compared to the DMBA group. Our study demonstrated the development of the novel class of fasudil derivatives potent anti-BC agent that improves intestinal flora and intestinal barrier function in rats.

Diagnostic value of cerebrospinal fluid levels of D-lactate, tumour necrosis factor-alpha and interleukin-6, -8, and -17 in suspected nosocomial meningitis.

This study aimed to determine the diagnostic value of interleukin (IL)-6, IL-8, IL-17, tumour necrosis factor-alpha (TNF-α) and D-lactate levels in the cerebrospinal fluid (CSF) of nosocomial meningitis patients. The CSF levels of cytokines and D-lactate were compared across 29 episodes of nosocomial meningitis, 38 episodes of pleocytosis (without meningitis) and 54 control subjects. The CSF levels of IL-6, IL-8, and D-lactate were higher in the group with nosocomial meningitis compared to the control group and the group with pleocytosis without meningitis ( P < 0.05). For IL-6 levels (threshold: >440 pg/mL), the sensitivity and specificity were 55.17% and 94.74%, respectively. For IL-8 levels (threshold: >1,249 pg/mL), the sensitivity and specificity were 44.83% and 84.21%, respectively. In patients with nosocomial meningitis, when the threshold of D-lactate levels was >1.05 μmol/mL, the sensitivity and specificity were 75.86% and 63.16%, respectively. In pleocytosis (without meningitis) CSF samples and in nosocomial meningitis CSF samples, the highest area under the receiver operating characteristic curve (AUC) was calculated for triple combination model of IL-6, IL-8 and D-lactate levels (AUC 0.801, P < 0.001) and double combination model of IL-6 and IL-8 (AUC 0.790, P < 0.001). Our study findings suggest that IL-6, IL-8 and D-lactate levels could be diagnostic markers for nosocomial meningitis.

Generation of a GLO-2 deficient mouse reveals its effects on liver carbonyl and glutathione levels

ObjectiveHydroxyacylglutathione hydrolase (aka as GLO-2) is a component of the glyoxalase pathway involved in the detoxification of the reactive oxoaldehydes, glyoxal and methylglyoxal. These reactive metabolites have been linked to a variety of pathological conditions, including diabetes, cancer and heart disease and may be involved in the aging process. The objective of this study was to generate a mouse model deficient in GLO-2 to provide insight into the function of GLO-2 and to determine if it is potentially linked to endogenous oxalate synthesis which could influence urinary oxalate excretion. MethodsA GLO-2 knock out mouse was generated using CRISPR/Cas 9 techniques. Tissue and 24-h urine samples were collected under baseline conditions from adult male and female animals for biochemical analyses, including chromatographic measurement of glycolate, oxalate, glyoxal, methylglyoxal, D-lactate, ascorbic acid and glutathione levels. ResultsThe GLO-2 KO animals developed normally and there were no changes in 24-h urinary oxalate excretion, liver levels of methylglyoxal, glyoxal, ascorbic acid and glutathione, or plasma d-lactate levels. GLO-2 deficient males had lower plasma glycolate levels than wild type males while this relationship was not observed in females. ConclusionsThe lack of a unique phenotype in a GLO-2 KO mouse model under baseline conditions is consistent with recent evidence, suggesting a functional glyoxalase pathway is not required for optimal health. A lower plasma glycolate in male GLO-2 KO animals suggests glyoxal production may be a significant contributor to circulating glycolate levels, but not to endogenous oxalate synthesis.

The Serum Citrulline and D-Lactate are Associated with Gastrointestinal Dysfunction and Failure in Critically Ill Patients.

ObjectiveThis study attempted to screen and combine effective biomarkers to analyse the association between these biomarkers and gastrointestinal failure (GIF) in critically ill patients.MethodsA total of 110 critically ill patients with acute gastrointestinal injury (AGI) admitted to ICU were enrolled. The AGI grade was determined by the AGI classification proposed by ESICM. There were 67 patients in gastrointestinal dysfunction (GID) group (AGI grade II), 43 patients in GIF group (AGI grade III–IV), and 41 healthy adults in healthy control (HC) group. APACHE II and SOFA score were used to evaluate the disease severity. Peripheral blood samples of patients were collected within 24 hours of admission to the ICU (prior-treatment) and after the conventional medication therapy for 7 consecutive days (post-treatment). Citrulline serum level was detected by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method, and D-lactate and lipopolysaccharide (LPS) serum levels were measured by ELISA. Pearson correlation, logistic regression, and ROC curve analysis were used.ResultsPatients with GID or GIF had lower serum level of citrulline, while higher levels of D-lactate and LPS than HC. Compared with GID patients, serum level of citrulline was reduced, while D-lactate and LPS were elevated in GIF patients. Correlation analysis displayed that serum levels of citrulline, D-lactate, and LPS were associated with the APACHE II and SOFA score in patients with GID or GIF. Logistics regression analysis showed that citrulline and D-lactate were risk for both GID and GIF. ROC curve analysis exhibited that combination of citrulline and D-lactate had relatively high value to distinguish GID from HC, GIF from GID, and GIF from HC.ConclusionSerum citrulline and D-lactate were associated with severity of GIF, combination of citrulline and D-lactate improved the diagnostic efficacy to identify GIF in critically ill patients.

Diverse expression patterns of mucin 2 in colorectal cancer indicates its mechanism related to the intestinal mucosal barrier

BACKGROUNDAbnormal expression patterns of mucin 2 (MUC2) have been reported in a variety of malignant tumors and precancerous lesions. Reduced MUC2 expression in the intestinal mucosa, caused by various pathogenic factors, is related to mechanical dysfunction of the intestinal mucosa barrier and increased intestinal mucosal permeability. However, the relationship between MUC2 and the intestinal mucosal barrier in patients with colorectal cancer (CRC) is not clear. AIMTo explore the relationship between MUC2 and intestinal mucosal barrier by characterizing the multiple expression patterns of MUC2 in CRC.METHODSImmunohistochemical staining was performed on intestinal tissue specimens from 100 CRC patients, including both cancer tissues and adjacent normal tissues. Enzyme-linked immunosorbent assays were performed on preoperative sera from 66 CRC patients and 20 normal sera to detect the serum levels of MUC2, diamine oxide (DAO), and D-lactate (D-LAC). The relationship between MUC2 expression and clinical parameters was calculated by the χ2 test or Fisher’s exact test. Prognostic value of MUC2 was evaluated by Kaplan-Meier curve and log-rank tests.RESULTSImmunohistochemical staining of 100 CRC tissues showed that the expression of MUC2 in cancer tissues was lower than that in normal tissues (54% vs 79%, P < 0.05), and it was correlated with tumor-node-metastasis (TNM) stage and lymph node metastasis in CRC patients (P < 0.05). However, the serum level of MUC2 in CRC patients was higher than that in normal controls, and was positively associated with serum levels of human DAO (χ2 = 3.957, P < 0.05) and D-LAC (χ2 = 7.236, P < 0.05), which are the biomarkers of the functional status of the intestinal mucosal barrier. And the serum level of MUC2 was correlated with TNM stage, tumor type, and distant metastasis in CRC patients (P < 0.05). Kaplan-Meier curves showed that decreased MUC2 expression in CRC tissues predicted a poor survival.CONCLUSIONMUC2 in tissues may play a protective role by participating in the intestinal mucosal barrier and can be used as an indicator to evaluate the prognosis of CRC patients.

Systemic inflammation down-regulates glyoxalase-1 expression: an experimental study in healthy males.

Background: Hypoxia and inflammation are hallmarks of critical illness, related to multiple organ failure. A possible mechanism leading to multiple organ failure is hypoxia- or inflammation-induced down-regulation of the detoxifying glyoxalase system that clears dicarbonyl stress. The dicarbonyl methylglyoxal (MGO) is a highly reactive agent produced by metabolic pathways such as anaerobic glycolysis and gluconeogenesis. MGO leads to protein damage and ultimately multi-organ failure. Whether detoxification of MGO into D-lactate by glyoxalase functions appropriately under conditions of hypoxia and inflammation is largely unknown. We investigated the effect of inflammation and hypoxia on the MGO pathway in humans in vivo.Methods: After prehydration with glucose 2.5% solution, ten healthy males were exposed to hypoxia (arterial saturation 80–85%) for 3.5 h using an air-tight respiratory helmet, ten males to experimental endotoxemia (LPS 2 ng/kg i.v.), ten males to LPS+hypoxia and ten males to none of these interventions (control group). Serial blood samples were drawn, and glyoxalase-1 mRNA expression, MGO, methylglyoxal-derived hydroimidazolone-1 (MG-H1), D-lactate and L-lactate levels, were measured serially.Results: Glyoxalase-1 mRNA expression decreased in the LPS (β (95%CI); -0.87 (-1.24; -0.50) and the LPS+hypoxia groups; -0.78 (-1.07; -0.48) (P<0.001). MGO was equal between groups, whereas MG-H1 increased over time in the control group only (P=0.003). D-Lactate was increased in all four groups. L-Lactate was increased in all groups, except in the control group.Conclusion: Systemic inflammation downregulates glyoxalase-1 mRNA expression in humans. This is a possible mechanism leading to cell damage and multi-organ failure in critical illness with potential for intervention.