Serology and molecular genetic analysis of two unrelated individuals with the same novel CisAB blood type.

OBJECTIVE: This study aims to report two unrelated individuals with the same novel CisAB blood type and confirm this rare blood type using a comprehensive approach that combines serological and molecular biology techniques. METHODS: Peripheral blood samples were collected from two patients and their family members. ABO blood typing and antibody detection were performed using conventional tube methods. Molecular biology techniques were employed to amplify and sequence the 6th and 7th exons of the ABO gene, with reference to gene mutation databases provided by NCBI and ISBT. RESULTS: The genotypes of the two unrelated individuals were identical and were confirmed as a new genotype through ISBT gene database comparison. Serological testing results showed different antigen reaction patterns, especially in terms of reverse typing. Gene sequencing identified a series of mutation points, and both unrelated individuals and one of their daughters had mutations at 297 A>G, 526 C>G, 657 C>T, 703 G>A, 803 G>C, and 930 G>A. According to the comprehensive results from The Blood Group Antigen Gene Mutation Database provided by NCBI, the genotype was determined as Bw37. However, based on the results from Names for ABO (ISBT 001) blood group alleles v1.1 171023, the sequencing results indicated a novel mutation combination not found in the ISBT database. Considering the serological reactions of all three individuals, the final determination was CisAB. CONCLUSIONS: This study confirmed the novel CisAB blood type in two individuals through the comprehensive application of serology and molecular biology techniques. The identified gene mutation points were not recorded in known databases, emphasizing the uniqueness of CisAB blood types. This research provides important insights into the genetic basis of ABO subtypes and the characteristics of CisAB blood types, and the relevant results have been submitted to the ISBT website for further research.

Assessing the impact of non-nutritive sweeteners on anthropometric indices and leptin levels in adults: A GRADE-assessed systematic review, meta-analysis, and meta-regression of randomized clinical trials.

In today's world, non-nutritive sweeteners (NNSs) are recognized as substitutes for sugar or other high-calorie sweeteners, and their consumption is increasing dramatically. However, there is ongoing debate regarding the impact of NNSs on anthropometric indices. To fill this gap in knowledge, the current GRADE-assessed systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted to evaluate the effects of artificial- and stevia-based sweeteners consumption on anthropometric indices and serum leptin level which is known as an appetite-regulating hormone. A comprehensive search was conducted on the Scopus, PubMed, and Embase databases up to November 2022 to identify randomized controlled trials (RCTs) investigating the effects of NNSs on anthropometric indices and serum leptin levels. Data extraction from qualified studies was performed independently by two researchers. A random- or fixed-effects model was used to estimate weighted mean differences (WMDs) and 95% confidence intervals (CIs) for anthropometric indices such as body weight (BW), body mass index (BMI), fat mass (FM), fat-free mass (FFM), waist circumference (WC) and serum leptin level. Heterogeneity between studies was assessed using Cochran's Q test and quantified using the I2 statistic. From a pool of 3212 studies initially identified, 20 studies with a total sample size of 2158 subjects were included in the analysis. Results of the pooled analysis showed that NNSs consumption had a significant reducing effect on BW (WMD: -1.02, 95% CI: -1.57, -0.46 Kg), FM (WMD: -1.09, 95% CI: -1.90, -0.29), and FFM (WMD: -0.83, 95% CI: -1.42, -0.23), but did not have any significant effect on BMI (WMD: -0.16, 95% CI: -0.35, 0.02), WC (WMD: -1.03, 95% CI: -2.77, 0.72), or serum leptin level (WMD: -2.17, 95% CI: -4.98, 0.65). The findings of this study indicate that the consumption of artificial- and stevia-based sweeteners may lead to a reduction in body weight, fat mass, and free fat mass.

Effect of third trimester maternal vitamin D levels on placental weight to birth weight ratio in uncomplicated pregnancies.

OBJECTIVES: Vitamin D has critical role for the fetal and placental development. Today, placental weight (PW), fetal birth weight (BW), and the PW/BW ratio are used as markers of fetal development. The aim of this study is to evaluate the relationship between vitamin D levels and these markers in uncomplicated pregnancies. METHODS: This study included 108 women with uncomplicated pregnancies, defined as full-term and healthy pregnancies without perinatal complications. Vitamin D levels <12 ng/mL were classified as deficient, 12-20 ng/mL as insufficient, and >20 ng/mL as normal. Postnatal BW and PW were compared according to maternal serum vitamin D levels. RESULTS: Maternal age, maternal height, maternal weight, body mass index, nulliparity, gestational age at delivery, mode of delivery, and fetal gender were similar between groups. Postnatal BW, PW, fetal height at birth, and fetal head circumference parameters were similar between the groups. The PW/BW ratio was 21.77±2.20 in the vitamin D deficient group, 21.20±2.40 in the insufficient group, and 19.98±2.37 in the normal group (p=0.012). In addition, there was a significant negative correlation between vitamin D level and the PW/BW ratio (p=0.012, r=0.031). CONCLUSIONS: Our results indicated that PW/BW ratio which is the marker for prediction adverse perinatal outcomes were significantly increased in the presence of vitamin D deficiency and insufficiency.

Neuropeptide B (NPB) and NPB receptor 2b (NPBWR2b) in the ricefield eel Monopterus albus: expression and potential involvement in the regulation of gonadotropins.

The neuropeptide B/W signaling system is composed of neuropeptide B (NPB), neuropeptide W (NPW), and two cognate receptors, NPBWR1 and NPBWR2, which are involved in diverse physiological processes, including the central regulation of neuroendocrine axes in vertebrates. The components of this signaling system are not well conserved during vertebrate evolution, implicating its functional diversity. The present study characterized the ricefield eel neuropeptide B/W system, generated a specific antiserum against the neuropeptide B/W receptor, and examined the potential roles of the system in the regulation of adenohypophysial functions. The ricefield eel genome contains npba, npbb, and npbwr2b but lacks the npw, npbwr1, and npbwr2a genes. The loss of npw and npbwr1 probably occurred at the base of ray-finned fish radiation and that of npbwr2a species specifically in ray-finned fish. Npba and npbb genes are produced through whole-genome duplication (WGD) in ray-finned fish. The ricefield eel npba was expressed in the brain and some peripheral tissues, while npbb was predominantly expressed in the brain. The ricefield eel npbwr2b was also expressed in the brain and in some peripheral tissues, such as the pituitary, gonad, heart, and eye. Immunoreactive Npbwr2b was shown to be localized to Lh and Fsh cells but not to Gh or Prl cells in the pituitary of ricefield eels. Npba upregulated the expression of fshb and cga but not lhb mRNA in pituitary fragments of ricefield eels cultured in vitro. The results of the present study suggest that the NPB system of ricefield eels may be involved in the neuroendocrine regulation of reproduction.

Effects of delta-opioid receptor agonist pretreatment on the cardiotoxicity of bupivacaine in rats.

BACKGROUND: Delta-opioid receptor is widely expressed in human and rodent hearts, and has been proved to protect cardiomyocytes against ischemia/reperfusion and heart failure. The antagonist of delta-opioid receptor could block the rescue effect of lipid emulsion against local anesthetic cardiotoxicity. However, no evidence is available for the direct effect of delta-opioid-receptor agonists on the cardiotoxicity of local anesthetics. METHODS: Anesthetized Sprague Dawley rats were divided into five groups. Group NS received 2 ml·kg-1·min-1 normal saline, group LE received 2 ml·kg-1·min-1 30% lipid emulsion and group BW received 0.1, 1.0, or 5.0 mg/kg BW373U86, a delta-opioid-receptor agonist, for 5 min. Then 0.5% bupivacaine was infused intravenously at a rate of 3.0 mg·kg-1·min-1 until asystole. The time of arrhythmia, 50% mean arterial pressure-, 50% heart rate-reduction and asystole were recorded, and the dose of bupivacaine at each time point was calculated. RESULTS: All three different doses of BW373U86 did not affect the arrhythmia, 50% mean arterial pressure-reduction, 50% heart rate-reduction and asystole dose of bupivacaine compared with group NS. 30% LE significantly increased the bupivacaine threshold of 50% mean arterial pressure-reduction (17.9 [15.4-20.7] versus 7.2 [5.9-8.7], p = 0.018), 50% heart rate-reduction (18.7 ± 4.2 versus 8.8 ± 1.7, p < 0.001) and asystole (26.5 [21.0-29.1] versus 11.3 [10.7-13.4], p = 0.008) compared with group NS. There was no difference between group LE and group NS in the arrhythmia dose of bupivacaine (9.9 [8.9-11.7] versus 5.6 [4.5-7.0], p = 0.060). CONCLUSIONS: Our data show that BW373U86 does not affect the cardiotoxicity of bupivacaine compared with NS control in rats. 30% LE pretreatment protects the myocardium against bupivacaine-induced cardiotoxicity.

Fetal Weight Estimation Using Automated Fractional Limb Volume With 2-Dimensional Size Parameters in Diabetic Pregnancies.

OBJECTIVES: To examine the effect of adding automated fetal fractional limb volume (FLV) with conventional 2-dimensional (2D) fetal weight estimation procedures in a cohort of diabetic pregnancies. METHODS: A pilot study of diabetic pregnancies measured standard fetal biometry within 7 days of delivery. Fractional arm volume (AVol) and fractional thigh volume (TVol) soft tissue parameters were measured with a commercially available automated software utility (5D Limb Vol; Samsung Medison Co, Ltd, Seoul, Korea). Three conventional weight prediction models that included only 2D size parameters were compared to FLV models that included AVol or TVol. Estimated and actual birth weight (BW) were assessed for the mean percent difference ± standard deviation of the percent differences. Systematic errors were evaluated by the Student t test, and random errors were compared by the Pitman test for correlated variances. The proportion of neonates with estimated fetal weight within 10% of BW was compared between groups by the McNemar test. RESULTS: Ninety gravid women were enrolled with pregestational (26.7%) or gestational (73.3%) diabetes. All prediction models were accurate, with the exception of small underestimations by the model of Schild et al (-3.8%; Ultrasound Obstet Gynecol 2004; 23:30-35). Random errors for the AVol (6.2%) and TVol (6.9%) models were significantly more precise than the other 3 prediction models: Hadlock et al (7.8%; Am J Obstet Gynecol 1985; 151:333-337), INTERGROWTH-21st (8.0%; Ultrasound Obstet Gynecol 2017; 49:478-486), and Schild et al (8.6%; P < .01). The greatest proportion of cases with BW ±10% was also classified by the AVol (91.1%) and TVol (91.1%) models, followed by Hadlock (83.3%), INTERGROWTH-21st (78.9%), and Schild (76.7%) predictions. CONCLUSIONS: The addition of automated FLV measurements to conventional 2D biometry was associated with improved weight predictions in this cohort of diabetic pregnancies.

Birthweight predicts glomerular filtration rate in adult-life: population based cross sectional study.

INTRODUCTION: Worldwide, there is a global progressive rise of chronic kidney disease. In parallel, children born after intra-uterine growth retardation are surviving to adult-life and beyond. This study describes the association of birthweight with and estimated glomerular filtration rate (eGFR). METHODS: Australian Diabetes, Obesity and Lifestyle (AusDiab) study participants were asked to complete a birthweight questionnaire. The associations between birthweight and eGFR were determined. RESULTS: A total of 4502 reported information related to their birthweight, with the other responders did not provide a value. The birthweight of the participants ranged from 0.4 to 7.0 kg with a mean-(SD) of 3.37 (0.7) kg. The mean (95%CI) birthweight was lower for females, 3.28 (0.6) kg, when compared to males, 3.5 (0.7) kg. Eight percent had a birthweight less than 2.5 kg. The eGFR was strongly and positively associated with birthweight, with people in the lowest sex-specific birthweight-quintiles having the lowest mean eGFR. This relationship persisted with adjustment for confounding factors. The OR(CI) for eGFR <10th-percentile (<61.4 ml/min for females and <73.4 for males) for people in the lowest vs. the higher birthweight-quintile was 2.19 (95%CI 1.14-4.2) for females and 2.37 (1.1-5.3) for males, after adjustment for other factors. CONCLUSIONS: Birthweight had a positive relationship with eGFR. Possible explanations include an association of birthweight with nephron-endowment. From a global health perspective but more in developing countries and in populations in epidemiologic transition, where substantially lower birthweights coexist with recently improved infant and adult survivals, the overall impact of this phenomenon on the population health profile could be more substantial.

Novel Hydrogen Sulfide (H2S)-Releasing BW-HS-101 and Its Non-H2S Releasing Derivative in Modulation of Microscopic and Molecular Parameters of Gastric Mucosal Barrier.

Hydrogen sulfide (H2S) is an endogenously produced molecule with anti-inflammatory and cytoprotective properties. We aimed to investigate for the first time if a novel, esterase-sensitive H2S-prodrug, BW-HS-101 with the ability to release H2S in a controllable manner, prevents gastric mucosa against acetylsalicylic acid-induced gastropathy on microscopic and molecular levels. Wistar rats were pretreated intragastrically with vehicle, BW-HS-101 (0.5-50 µmol/kg) or its analogue without the ability to release H2S, BW-iHS-101 prior to ASA administration (125 mg/kg, intragastrically). BW-HS-101 was administered alone or in combination with nitroarginine (L-NNA, 20 mg/kg, intraperitoneally) or zinc protoporphyrin IX (10 mg/kg, intraperitoneally). Gastroprotective effects of BW-HS-101 were additionally evaluated against necrotic damage induced by intragastrical administration of 75% ethanol. Gastric mucosal damage was assessed microscopically, and gastric blood flow was determined by laser flowmetry. Gastric mucosal DNA oxidation and PGE2 concentration were assessed by ELISA. Serum and/or gastric protein concentrations of IL-1α, IL-1ß, IL-2, IL-4, IL-6, IL-10, IL-13, VEGF, GM-CSF, IFN-γ, TNF-α, and EGF were determined by a microbeads/fluorescent-based multiplex assay. Changes in gastric mucosal iNOS, HMOX-1, SOCS3, IL1-R1, IL1-R2, TNF-R2, COX-1, and COX-2 mRNA were assessed by real-time PCR. BW-HS-101 or BW-iHS-101 applied at a dose of 50 µmol/kg protected gastric mucosa against ASA-induced gastric damage and prevented a decrease in the gastric blood flow level. H2S prodrug decreased DNA oxidation, systemic and gastric mucosal inflammation with accompanied upregulation of SOCS3, and EGF and HMOX-1 expression. Pharmacological inhibition of nitric oxide (NO) synthase but not carbon monoxide (CO)/heme oxygenase (HMOX) activity by L-NNA or ZnPP, respectively, reversed the gastroprotective effect of BW-HS-101. BW-HS-101 also protected against ethanol-induced gastric injury formation. We conclude that BW-HS-101, due to its ability to release H2S in a controllable manner, prevents gastric mucosa against drugs-induced gastropathy, inflammation and DNA oxidation, and upregulate gastric microcirculation. Gastroprotective effects of this H2S prodrug involves endogenous NO but not CO activity and could be mediated by cytoprotective and anti-inflammatory SOCS3 and EGF pathways.

Role of astaxanthin in the modulation of brain-derived neurotrophic factor and spatial learning behavior in perinatally undernourished Wistar rats.

Objective: Maternal health and nutrition during the perinatal period is the predominant factor influencing the functional development of the brain. Maternal malnutrition during the perinatal period causes retardation of brain development. The current study investigates the role of Astaxanthin (AsX) in spatial learning and memory and BDNF in perinatally undernourished Wistar rats.Methods: The albino wistar rats were perinatally undernourished and administered with different dosages of AsX. The spatial learning and memory performance and BDNF level were assessed. Data were collected and analysed.Results: The % Correct choice during the acquisition phase, performance at the end of the acquisition phase and the mean BDNF level at the Hippocampus, Cerebellum, and Cerebral cortex showed significant decline (P<0.001) in the PUN group and significantly high (P<0.001) in the PUNA2 group compared to the control. However, the mean RME and mean WME during different days of the acquisition phase were significantly high (P<0.001) in the PUN group and insignificant (P>0.05) in PUNA2 compared to the control.Discussion: The results showed that AsX effectively modulated the cognitive deficit that occurred in perinatally undernourished rats. This can be attributed to BDNF upregulation as evidenced by the significant increase of the BDNF level.

Blocking properties of terguride at the 5-HT2 receptor subtypes mediating cardiovascular responses in the rat.

In vitro studies have suggested that terguride blocks the contractile and relaxant responses produced by 5-hydroxytryptamine (5-HT) via 5-HT2A/2B receptors. This study has now investigated terguride's blocking properties on central/peripheral 5-HT2 receptors in anaesthetized or pithed rats. Male Wistar anaesthetized/pithed rats were cannulated for recording blood pressure and heart rate and for i.v. administration of several compounds. In both groups of rats, i.v. bolus injections of 5-HT or (±)-DOI (a 5-HT2 receptor agonist; 1-1000 µg/kg) produced dose-dependent increases in diastolic blood pressure and heart rate. These responses were dose-dependently antagonized by terguride (10-3000 µg/kg). In anaesthetized rats, i.v. bolus injections of BW723C86 (a 5-HT2B receptor agonist; 1-1000 µg/kg) produced dose-dependent increases in diastolic blood pressure and not dose-dependent increases in heart rate, while in pithed rats, these responses were attenuated. The vasopressor responses elicited by BW723C86 in anaesthetized rats were dose-dependently blocked by terguride (10-300 µg/kg), whereas its the tachycardic responses were dose-independently blocked. These results, taken together, suggest that terguride behaved as an antagonist at the 5-HT2 receptors located in the central nervous system and (or) the systemic vasculature. This is the first evidence demonstrating that terguride can block central/peripheral 5-HT2 receptors mediating cardiovascular responses in anaesthetized or pithed rats.

BW373U86 upregulates autophagy by inhibiting the PI3K/Akt pathway and regulating the mTOR pathway to protect cardiomyocytes from hypoxia-reoxygenation injury.

The purpose of this study was to explore the protective effect of BW373U86 (a δ-opioid receptor (DOR) agonist) on ischemia-reperfusion (I/R) injury in rat cardiomyocytes and its underlying mechanism. Primary rat cardiomyocytes were cultured and pretreated with BW373U86 for intervention. The cardiomyocytes were cultured under the condition of 94% N2 and 5% CO2 for 24 h to perform hypoxia culture and conventionally cultured for 12 h to perform reoxygenation culture. The cell viability of cardiomyocytes was detected by an MTT assay (Sigma-Aldrich). The autophagy lysosome levels in cardiomyocytes were evaluated by acidic vesicular organelles with dansylcadaverine (MDC) staining (autophagy test kit, Kaiji Biology, kgatg001). The protein expression levels of LC3, p62, and factors in the PI3K/Akt/mTOR signaling pathway were detected by Western blot. Pretreatment with BW373U86 could improve the cell viability of cardiomyocytes with hypoxia-reoxygenation (H/R) injury (p < 0.05). Interestingly, after coculture of BW373U86 and PI3K inhibitor (3-methyladenine), the protein expression levels of p-Akt in cardiomyocytes were markedly increased in comparison with those in the BW373U86 group (p < 0.05). However, there were no significant differences in the protein expression levels of mTOR between the coculture group and the BW373U86 group (p > 0.05). BW373U86 upregulated autophagy to protect cardiomyocytes from H/R injury, which may be related to the PI3K/Akt/m TOR pathway.

CLONAL SPREAD OF YERSINIA ENTEROCOLITICA 1B/O:8 IN MULTIPLE ZOO SPECIES.

Yersinia enterocolitica (YE) bioserotype 1B/O:8 (YE 1B/O:8) was identified in routine culture of a variety of zoo species housed at Omaha's Henry Doorly Zoo and Aquarium (OHDZA) from April to July 2011. Animal cases representing 12 species had YE detected from 34 cases during routine fecal monitoring and/or during postmortem examination: Coquerel's sifakas (Propithecus coquereli, two cases), black & white (BW) ruffed lemurs (Varecia variegata variegata, six cases), red ruffed lemurs (Varecia rubra, seven cases), white handed gibbon (Hylobates lar albimana, one case), black lemurs (Eulemur macaco, three cases), mongoose lemurs (Eulemur mongoz, two cases), African hunting dogs (Lycaon pictus, five cases), agile gibbons (Hylobates agilis, three cases), siamangs (Hylobates syndactylus, two cases), colobus monkey (Colobus angolensis palliates, one case), argus pheasant (Argusianus argus, one case), and orangutan (Pongo pygmaeus, one case). Most species were not symptomatic; however, three symptomatic cases in Coquerel's sifakas (two) and a white handed gibbon (one) showed clinical signs of diarrhea and lethargy that resulted in death for the Coquerel's sifakas. One unexpected death also occurred in a BW ruffed lemur. To the authors' knowledge, this is the first report of YE 1B/O:8 in such a large variety of zoo species. The source of the YE could not be identified, prompting the initiation of a diseases surveillance program to prevent further cases for the species that are sensitive to YE. To date, no additional cases have been identified, thus suggesting a single introduction of the YE 1B/O:8 strain into the zoo environment.

The association between dietary protein intake, energy intake and physical frailty: results from the Rotterdam Study.

Sufficient protein intake has been suggested to be important for preventing physical frailty, but studies show conflicting results which may be explained because not all studies address protein source and intake of other macronutrients and total energy. Therefore, we studied 2504 subjects with data on diet and physical frailty, participating in a large population-based prospective cohort among subjects aged 45+ years (the Rotterdam Study). Dietary intake was assessed with a FFQ. Frailty was defined according to the frailty phenotype as the presence of at least three out of the following five symptoms: weight loss, low physical activity, weakness, slowness and fatigue. We used multinomial logistic regression models to evaluate the independent association between protein intake and frailty using two methods: nutrient residual models and energy decomposition models. With every increase in 10 g total, plant or animal protein per d, the odds to be frail were 1·06 (95 % CI 0·98, 1·15), 0·87 (95 % CI 0·71, 1·07) and 1·07 (95 % CI 0·99, 1·15), respectively, using the nutrient residual method. Using the energy partition model, we observed that the odds to be frail were lower with higher vegetable protein intake (OR per 418·4 kJ (100 kcal): 0·61, 95 % CI 0·39, 0·97), however, results disappeared when adjusting for physical activity. For energy intake from any source we observed that with every 418·4 kJ (100 kcal) increase, the odds to be frail were 5 % lower (OR: 0·95, 95 % CI 0·93, 0·97). Our results suggest that energy intake, but not protein specifically, is associated with less frailty. Considering other macronutrients, physical activity and diet quality seems to be essential for future studies on protein and frailty.

Fluoride retention in infants living in fluoridated and non-fluoridated areas: effects of weaning.

Limited knowledge is available on total fluoride exposure, excretion and retention in infants, despite the first year of human life being the critical period for dental development and risk of dental fluorosis. This study investigated total daily fluoride intake (TDFI), excretion (TDFE) and retention (TDFR) in infants living in fluoridated and non-fluoridated water areas at pre- and post-weaning stages of development. Healthy infants, aged 0-12 months, were recruited and their TDFI (mg/kg body weight (BW) per d), from diet and toothpaste ingestion, was assessed over a 3-d period using a dietary diary and tooth-brushing questionnaire. TDFE (mg/kg BW per d) was estimated by collecting 48-h urine and faeces. TDFR (mg/kg BW per d) was estimated by subtracting TDFE from TDFI. A total of forty-seven infants completed the study: sixteen at pre-weaning and thirty-one at post-weaning stages, with a mean age of 3·4 and 10·0 months, respectively. TDFI was lower in the non-fluoridated area (P<0·001) and at the pre-weaning stage (P=0·002) but higher in formula-fed infants (P<0·001). TDFE was mainly affected by type of feeding, with higher excretion in formula-fed infants (P<0·001). TDFR was lower in the non-fluoridated area (P<0·001) and at the pre-weaning stage (P<0·001) but higher in formula-fed infants (P=0·001). In conclusion, a relatively large proportion of fluoride intake is retained in the body in weaned infants. This is an important consideration in fluoride-based prevention programmes, with goals to maximise caries prevention while minimising the risk of dental fluorosis.

Interactive effects of dietary fibre and lipid types modulate gastrointestinal flows and apparent digestibility of fatty acids in growing pigs.

A total of eight ileal and caecal cannulated Yorkshire barrows were used to determine the interactions of dietary fibre (DF) and lipid types on apparent digestibility of DM and fatty acids (FA) and FA flows in gastrointestinal segments. Pigs were offered four diets that contained either pectin or cellulose with or without beef tallow or maize oil in two Youden square designs (n 6). Each period lasted 15 d. Faeces, ileal and caecal contents were collected to determine apparent ileal digestibility (AID), apparent caecal digestibility and apparent total tract digestibility (ATTD) of dietary components. The interactions between DF and lipid types influenced (P <0·05) the digestibility of DM and FA flows. The addition of maize oil decreased (P <0·05) AID of DM in pectin diets, and the addition of beef tallow depressed (P <0·001) ATTD of DM in cellulose diets. Dietary supplementation with beef tallow decreased (P <0·05) the AID of FA in pectin-containing diets but had no effects in cellulose-containing diets. Dietary supplementation with beef tallow increased (P <0·05) AID of SFA and PUFA and the flow of ileal oleic, vaccenic, linolenic and eicosadienoic acids and reduced the flow of faecal lauric, docosatetraenoic and docosapentaenoic acids in pectin- and cellulose-containing diets. In conclusion, the interaction between DF type and lipid saturation modulates digestibility of DM and lipids and FA flows but differs for soluble and insoluble fibre sources, SFA and unsaturated fatty acids and varies in different gastrointestinal segments.

Effect of pre-exercise ingestion of α-lactalbumin on subsequent endurance exercise performance and mood states.

This study investigated the effect of pre-exercise α-lactalbumin ingestion on subsequent endurance exercise performance, muscle pain and mood states. In a two-stage cross-over counterbalance design, eleven male endurance runners (age: 31 (se 2) years, height: 169·5 (se 4·4) cm, weight: 63·6 (se 5·1) kg, V̇O2max: 58·8 (se 6·3) ml/kg per min) consumed two solutions (carbohydrate+α-lactalbumin, CA; carbohydrate+whey protein isolate, CW) 2 h before a self-paced 21-km run. Creatine kinase, IL-6, muscle pain, pressure pain threshold (PPT) and mood states were assessed 2 h before exercise, immediately before exercise (Pre-ex0) and immediately after exercise (Post-ex0). No difference was found in 21-km running performance between two trials (CA v. CW: 115·85 (se 5·20) v. 118·85 (se 5·51) min, P=0·48). Compared with CW, CA led to higher PPT at Pre-ex0 (41·77 (se 2·27) v. 35·56 (se 2·10) N/cm2, P<0·01) and Post-ex0 (38·76 (se 3·23) v. 35·30 (se 3·55) N/cm2, P=0·047). Compared with CW, CA reduced the feeling of fatigue at Post-ex0 (P<0·01); CA also reduced salivary cortisol levels at Post-ex0 (0·72 (se 0·07) v. 0·83 (se 0·13) ng/ml, P<0·01). In conclusion, the ingestion of α-lactalbumin did not improve the 21-km time-trial performance. However, compared with the pre-exercise ingestion of whey protein, that of α-lactalbumin led to superior results during similar levels of endurance exercise: it elevated PPT and reduced the feeling of fatigue and the cortisol levels.

The effect of chronic oral vitamin D supplementation on adiposity and insulin secretion in hypothalamic obese rats.

Reduced plasma vitamin D (VD) levels may contribute to excessive white adipose tissue, insulin resistance (IR) and dyslipidaemia. We evaluated the effect of chronic oral VD supplementation on adiposity and insulin secretion in monosodium glutamate (MSG)-treated rats. During their first 5 d of life, male neonate rats received subcutaneous injections of MSG (4 g/kg), while the control (CON) group received saline solution. After weaning, groups were randomly distributed into VD supplemented (12 µg/kg; three times/week) and non-supplemented (NS) rats, forming four experimental groups (n 15 rats/group): CON-NS, CON-VD, MSG-NS and MSG-VD. At 76 d of life, rats were submitted to an oral glucose tolerance test (OGTT; 2 g/kg), and at 86 d, obesity, IR and plasma metabolic parameters were evaluated. Pancreatic islets were isolated for glucose-induced insulin secretion (GIIS), cholinergic insulinotropic response and muscarinic 3 receptor (M3R), protein kinase C (PKC) and protein kinase A (PKA) expressions. Pancreas was submitted to histological analyses. VD supplementation decreased hyperinsulinaemia (86 %), hypertriacylglycerolaemia (50 %) and restored insulin sensibility (89 %) in MSG-VD rats, without modifying adiposity, OGTT or GIIS, compared with the MSG-NS group. The cholinergic action was reduced (57 %) in islets from MSG-VD rats, without any change in M3R, PKA or PKC expression. In conclusion, chronic oral VD supplementation of MSG-obese rats was able to prevent hyperinsulinaemia and IR, improving triacylglycerolaemia without modifying adiposity. A reduced cholinergic pancreatic effect, in response to VD, could be involved in the normalisation of plasma insulin levels, an event that appears to be independent of M3R and its downstream pathways.

Effect of lifelong carnitine supplementation on plasma and tissue carnitine status, hepatic lipid metabolism and stress signalling pathways and skeletal muscle transcriptome in mice at advanced age.

While strong evidence from clinical studies suggests beneficial effects of carnitine supplementation on metabolic health, serious safety concerns associated with carnitine supplementation have been raised from studies in mice. Considering that the carnitine doses in these mice studies were up to 100 times higher than those used in clinical studies, the present study aimed to address possible safety concerns associated with long-term supplementation of a carnitine dose used in clinical trials. Two groups of NMRI mice were fed either a control or a carnitine-supplemented diet (1 g/kg diet) from weaning to 19 months of age, and parameters of hepatic lipid metabolism and stress signalling and skeletal muscle gene expression were analysed in the mice at 19 months of age. Concentrations of free carnitine and acetylcarnitine in plasma and tissues were higher in the carnitine than in the control group (P<0·05). Plasma concentrations of free carnitine and acetylcarnitine were higher in mice at adult age (10 and 15 months) than at advanced age (19 months) (P<0·05). Hepatic mRNA and protein levels of genes involved in lipid metabolism and stress signalling and hepatic and plasma lipid concentrations did not differ between the carnitine and the control group. Skeletal muscle transcriptome analysis in 19-month-old mice revealed only a moderate regulation between carnitine and control group. Lifelong carnitine supplementation prevents an age-dependent impairment of plasma carnitine status, but safety concerns associated with long-term supplementation of carnitine at doses used in clinical trials can be considered as unfounded.

Starch digestion kinetics and mechanisms of hydrolysing enzymes in growing pigs fed processed and native cereal-based diets.

This study aimed to examine in vivo starch digestion kinetics and to unravel the mechanisms of starch hydrolysing enzymes. Ninety pigs (23 (sd 2·1) kg body weight) were assigned to one of nine treatments in a 3×3 factorial arrangement, with starch source (barley, maize, high-amylose (HA) maize) and form (isolated, within cereal matrix, extruded) as factors. We determined starch digestion coefficients (DC), starch breakdown products and digesta retention times in four small-intestinal segments (SI1-4). Starch digestion in SI2 of pigs fed barley and maize, exceeded starch digestion of pigs fed HA maize by 0·20-0·33 DC units (P<0·01). In SI3-4, barley starch were completely digested, whereas the cereal matrix of maize hampered digestion and generated 16 % resistant starch in the small intestine (P<0·001). Extrusion increased the DC of maize and HA maize starch throughout the small intestine but not that of barley (P<0·05). Up to 25 % of starch residuals in the proximal small intestine of pigs was present as glucose and soluble α(1-4) maltodextrins. The high abundance of glucose, maltose and maltotriose in the proximal small intestine indicates activity of brush-border enzymes in the intestinal lumen, which is exceeded by α-amylase activity. Furthermore, we found that in vivo starch digestion exceeded our in vitro predictions for rapidly digested starch, which indicates that the role of the stomach on starch digestion is currently underestimated. Consequently, in vivo glucose release of slowly digestible starch is less gradual than expected, which challenges the prediction quality of the in vitro assay.

Acute dietary zinc deficiency in rats exacerbates myocardial ischaemia-reperfusion injury through depletion of glutathione.

Zn plays an important role in maintaining the anti-oxidant status within the heart and helps to counter the acute redox stress that occurs during myocardial ischaemia and reperfusion. Individuals with low Zn levels are at greater risk of developing an acute myocardial infarction; however, the impact of this on the extent of myocardial injury is unknown. The present study aimed to compare the effects of dietary Zn depletion with in vitro removal of Zn (N,N,N',N'-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN)) on the outcome of acute myocardial infarction and vascular function. Male Sprague-Dawley rats were fed either a Zn-adequate (35 mg Zn/kg diet) or Zn-deficient (<1 mg Zn/kg diet) diet for 2 weeks before heart isolation. Perfused hearts were subjected to a 30 min ischaemia/2 h reperfusion (I/R) protocol, during which time ventricular arrhythmias were recorded and after which infarct size was measured, along with markers of anti-oxidant status. In separate experiments, hearts were challenged with the Zn chelator TPEN (10 µm) before ischaemia onset. Both dietary and TPEN-induced Zn depletion significantly extended infarct size; dietary Zn depletion was associated with reduced total cardiac glutathione (GSH) levels, while TPEN decreased cardiac superoxide dismutase 1 levels. TPEN, but not dietary Zn depletion, also suppressed ventricular arrhythmias and depressed vascular responses to nitric oxide. These findings demonstrate that both modes of Zn depletion worsen the outcome from I/R but through different mechanisms. Dietary Zn deficiency, resulting in reduced cardiac GSH, is the most appropriate model for determining the role of endogenous Zn in I/R injury.