Metabolic characteristics of transmembrane prolyl 4-hydroxylase (P4H-TM) deficient mice.

Transmembrane prolyl 4-hydroxylase (P4H-TM) is an enigmatic enzyme whose cellular function and primary substrate remain to be identified. Its loss-of-function mutations cause a severe neurological HIDEA syndrome with hypotonia, intellectual disability, dysautonomia and hypoventilation. Previously, P4H-TM deficiency in mice was associated with reduced atherogenesis and lower serum triglyceride levels. Here, we characterized the glucose and lipid metabolism of P4h-tm-/- mice in physiological and tissue analyses. P4h-tm-/- mice showed variations in 24-h oscillations of energy expenditure, VO2 and VCO2 and locomotor activity compared to wild-type (WT) mice. Their rearing activity was reduced, and they showed significant muscle weakness and compromised coordination. Sedated P4h-tm-/- mice had better glucose tolerance, lower fasting insulin levels, higher fasting lactate levels and lower fasting free fatty acid levels compared to WT. These alterations were not present in conscious P4h-tm-/- mice. Fasted P4h-tm-/- mice presented with faster hepatic glycogenolysis. The respiratory rate of conscious P4h-tm-/- mice was significantly lower compared to the WT, the decrease being further exacerbated by sedation and associated with acidosis and a reduced ventilatory response to both hypoxia and hypercapnia. P4H-TM deficiency in mice is associated with alterations in whole-body energy metabolism, day-night rhythm of activity, glucose homeostasis and neuromuscular and respiratory functions. Although the underlying mechanism(s) are not yet fully understood, the phenotype appears to have neurological origins, controlled by brain and central nervous system circuits. The phenotype of P4h-tm-/- mice recapitulates some of the symptoms of HIDEA patients, making this mouse model a valuable tool to study and develop tailored therapies.

Relationship between Ketones, Ghrelin, and, Appetite on Isocaloric Diets with Varying Carbohydrate Quality and Amount: Results from a Randomized Controlled Trial in People with Obesity (CARBFUNC).

BACKGROUND: Low-carbohydrate high-fat (LCHF) diets may suppress the increase in appetite otherwise seen after diet-induced fat loss. However, studies of diets without severe energy restriction are lacking, and the effects of carbohydrate quality relative to quantity have not been directly compared. OBJECTIVES: To evaluated short- (3 mo) and long-term (12 mo) changes in fasting plasma concentrations of total ghrelin, ß-hydroxybutyrate (ßHB), and subjective feelings of appetite on 3 isocaloric eating patterns within a moderate caloric range (2000-2500 kcal/d) and with varying carbohydrate quality or quantity. METHODS: We performed a randomized controlled trial of 193 adults with obesity, comparing eating patterns based on "acellular" carbohydrate sources (e.g., flour-based whole-grain products; comparator arm), "cellular" carbohydrate sources (minimally processed foods with intact cellular structures), or LCHF principles. Outcomes were compared by an intention-to-treat analysis using constrained linear mixed modeling. This trial was registered at clinicaltrials.gov as NCT03401970. RESULTS: Of the 193 adults, 118 (61%) and 57 (30%) completed 3 and 12 mo of follow-up. Throughout the intervention, intakes of protein and energy were similar with all 3 eating patterns, with comparable reductions in body weight (5%-7%) and visceral fat volume (12%-17%) after 12 mo. After 3 mo, ghrelin increased significantly with the acellular (mean: 46 pg/mL; 95% CI: 11, 81) and cellular (mean: 54 pg/mL; 95% CI: 21, 88) diets but not with the LCHF diet (mean: 11 pg/mL; 95% CI: -16, 38). Although ßHB increased significantly more with the LCHF diet than with the acellular diet after 3 m (mean: 0.16 mmol/L; 95% CI: 0.09, 0.24), this did not correspond to a significant group difference in ghrelin (unless the 2 high-carbohydrate groups were combined [mean: -39.6 pg/mL; 95% CI: -76, -3.3]). No significant between-group differences were seen in feelings of hunger. CONCLUSIONS: Modestly energy-restricted isocaloric diets differing in carbohydrate cellularity and amount showed no significant differences in fasting total ghrelin or subjective hunger feelings. An increase in ketones with the LCHF diet to 0.3-0.4 mmol/L was insufficient to substantially curb increases in fasting ghrelin during fat loss.

Correlation of Antithrombin Functional Status and Levels with Liver Fibrosis in Patients with Hepatitis C.

BACKGROUND: Abnormal levels of coagulation factors and their inhibitors have shown association with liver diseases. The objective of this study was to determine the qualitative and quantitive status of antithrombin (AT) in patients with chronic liver disease associated hepatitis C infection and their correlation with severity of liver fibrosis. METHODS: In this study, 75 (43 male and 32 female) known patients with chronic liver disease associated hepatitis C infection were enrolled. AT activity and quantitative immunoassays were carried out using Stachrom AT reagent kit (Diagnostica Stago, France) and Liatest AT reagent (Diagnostica Stago, France), respectively. Hepatic biopsies were obtained and graded for liver fibrosis from all study participants. RESULTS: Of the 75 patients, 45 had normal AT while 30 showed lower activity of AT. Similarly, the quantitative assay showed reduced levels of AT in 30 patients and normal levels in 45 patients. CONCLUSIONS: In the early stages of liver fibrosis, AT activity and antigenic levels were found to be normal or minimally affected. While advanced stages of the disease showed markedly reduced levels of AT and activity. Hence, it can be concluded that the degree of fibrosis affects the status of AT.

ER-stress promotes VHL-independent degradation of hypoxia-inducible factors via FBXW1A/ßTrCP.

Metabolic adaptation and signal integration in response to hypoxic conditions is mainly regulated by hypoxia-inducible factors (HIFs). At the same time, hypoxia induces ROS formation and activates the unfolded protein response (UPR), indicative of endoplasmic reticulum (ER) stress. However, whether ER stress would affect the hypoxia response remains ill-defined. Here we report that feeding mice a high fat diet causes ER stress and attenuates the response to hypoxia. Mechanistically, ER stress promotes HIF-1α and HIF-2α degradation independent of ROS, Ca2+, and the von Hippel-Lindau (VHL) pathway, involving GSK3ß and the ubiquitin ligase FBXW1A/ßTrCP. Thereby, we reveal a previously unknown function of the GSK3ß/HIFα/ßTrCP1 axis in ER homeostasis and demonstrate that inhibition of the HIF-1 and HIF-2 response and genetic deficiency of GSK3ß affects proliferation, migration, and sensitizes cells for ER stress promoted apoptosis. Vice versa, we show that hypoxia affects the ER stress response mainly through the PERK-arm of the UPR. Overall, we discovered previously unrecognized links between the HIF pathway and the ER stress response and uncovered an essential survival pathway for cells under ER stress.

Dairy products influence gut hormone secretion and appetite differently: A randomized controlled crossover trial.

Little is known about how dairy products with different nutrient contents and food matrices affect appetite sensation and gut hormone secretion. The objective of this study was to investigate how appetite sensation and gut hormone secretion in healthy adults are affected by meals with the same amount of fat but from different dairy products. Forty-seven healthy adults (70% women) were recruited to a randomized controlled crossover study with 4 dairy meals consisting of butter, cheese, whipped cream, or sour cream, corresponding to 45 g (approximately 60 energy percent) of fat. Plasma samples were collected for analysis of cholecystokinin (CCK), pancreatic polypeptide (PP), peptide YY (PYY), and ghrelin concentrations at 0, 2, 4, and 6 h after the meals and analyzed as the incremental area under the curve (iAUC0-6h) in a mixed model. Hunger, satiety, and appetite sensations were measured with a visual analog scale (VAS) immediately after finishing the meals and at 4 and 6 h postprandially. Intake of cheese induced a higher level of plasma PP-iAUC0-6h compared with butter or whipped cream, and a higher level of plasma CCK-iAUC0-6h compared with whipped cream. Intake of whipped cream increased VAS appetite at 4 h compared with cheese or sour cream, and at 6 h compared with cheese or butter. No significant meal effect was found for hunger, satiety, plasma PYY, or plasma ghrelin concentration. Intake of cheese increased postprandial plasma PP and CCK concentrations and decreased appetite compared with whipped cream but not with sour cream. These findings encourage further investigations of how different dairy products affect gut hormone secretion and appetite sensation.

Linking Dysregulated AMPK Signaling and ER Stress in Ethanol-Induced Liver Injury in Hepatic Alcohol Dehydrogenase Deficient Deer Mice.

Ethanol (EtOH) metabolism itself can be a predisposing factor for initiation of alcoholic liver disease (ALD). Therefore, a dose dependent study to evaluate liver injury was conducted in hepatic alcohol dehydrogenase (ADH) deficient (ADH-) and ADH normal (ADH+) deer mice fed 1%, 2% or 3.5% EtOH in the liquid diet daily for 2 months. Blood alcohol concentration (BAC), liver injury marker (alanine amino transferase (ALT)), hepatic lipids and cytochrome P450 2E1 (CYP2E1) activity were measured. Liver histology, endoplasmic reticulum (ER) stress, AMP-activated protein kinase (AMPK) signaling and cell death proteins were evaluated. Significantly increased BAC, plasma ALT, hepatic lipids and steatosis were found only in ADH- deer mice fed 3.5% EtOH. Further, a significant ER stress and increased un-spliced X-box binding protein 1 were evident only in ADH- deer mice fed 3.5% EtOH. Both strains fed 3.5% EtOH showed deactivation of AMPK, but increased acetyl Co-A carboxylase 1 and decreased carnitine palmitoyltransferase 1A favoring lipogenesis were found only in ADH- deer mice fed 3.5% EtOH. Therefore, irrespective of CYP2E1 overexpression; EtOH dose and hepatic ADH deficiency contribute to EtOH-induced steatosis and liver injury, suggesting a linkage between ER stress, dysregulated hepatic lipid metabolism and AMPK signaling.

Meals with Similar Fat Content from Different Dairy Products Induce Different Postprandial Triglyceride Responses in Healthy Adults: A Randomized Controlled Cross-Over Trial.

BACKGROUND: Postprandial lipemia is a risk factor for cardiovascular disease. Dairy products differ in nutrient content and food matrix, and little is known about how different dairy products affect postprandial triglyceride (TG) concentrations. OBJECTIVE: We investigated the effect of meals with similar amounts of fat from different dairy products on postprandial TG concentrations over 6 h in healthy adults. METHODS: A randomized controlled cross-over study was performed on 47 subjects (30% men), with median (25th-75th percentile) age of 32 (25-46) y and body mass index of 23.6 (21.0-25.8) kg/m2. Meals included 1 of butter, cheese, whipped cream, or sour cream, corresponding to 45 g of fat (approximately 60 energy%). Serum concentrations of TGs (primary outcome), and total cholesterol (TC), low density lipoprotein cholesterol (LDL cholesterol), high density lipoprotein cholesterol (HDL cholesterol), insulin, glucose, non-esterified fatty acids, and plasma glucose-dependent insulinotropic polypeptide (secondary outcomes) were measured before the meal and 2, 4, and 6 h postprandially. Incremental AUC (iAUC) was calculated for the responses, and data were analyzed using a linear mixed model. RESULTS: Sour cream induced a 61% larger TG-iAUC0-6 h compared to whipped cream (P < 0.001), a 53% larger TG-iAUC0-6 h compared to butter (P < 0.001), and a 23% larger TG-iAUC0-6 h compared to cheese (P = 0.05). No differences in TG-iAUC0-6 h between the other meals were observed. Intake of sour cream induced a larger HDL cholesterol-iAUC0-6 h compared to cheese (P = 0.01). Intake of cheese induced a 124% larger insulin iAUC0-6 h compared to butter (P = 0.006). No other meal effects were observed. CONCLUSIONS: High-fat meals containing similar amount of fat from different dairy products induce different postprandial effects on serum TGs, HDL cholesterol, and insulin in healthy adults. The potential mechanisms and clinical impact of our findings remain to be further elucidated. The study was registered at www.clinicaltrials.gov as NCT02836106.

Identification and Characterization of Colletotrichum Species Associated with Mango Anthracnose in Guangxi, China.

Mango (Mangifera indica) is widely grown across southern China, especially in the provinces of Guangxi, Hainan, Yunnan, Sichuan, and Taiwan. Guangxi itself has over 86,667 ha of mango production. The purpose of this study was to identify Colletotrichum species associated with mango in different parts of Guangxi and examine their pathogenicity on leaves and fruits of mango in vitro. Diseased leaves were collected from 25 mango orchards in different areas of Guangxi province. Sixty-five isolates were obtained from mango leaves with anthracnose symptoms, and these were further characterized based on morphology and DNA sequencing. Twenty-nine isolates from different areas were selected for sequencing and analyses of the internal transcribed spacer region, glyceraldehyde-3-phosphate dehydrogenase, partial actin, ß-tubulin, and chitin synthase genomic regions. The most common fungal isolates were these three species: Colletotrichum asianum, C. fructicola, and C. siamense. C. asianum was the most common and widely distributed in Guangxi (51.7%), followed by C. fructicola (37.9%) and C. siamense (10.2%), both found in Tiandong, Tianyang, and Wuming counties. There was no evidence of geographical specialization of the different species. Pathogenicity assays showed that all isolates were pathogenic to mango leaves and fruit (cultivar Tainong). No relationship was found between origin of isolates and their virulence. This is the first description of C. asianum, C. fructicola, and C. siamense as causal agents of mango leaf anthracnose from Guangxi province, China.

Whole genome re-sequencing of date palms yields insights into diversification of a fruit tree crop.

Date palms (Phoenix dactylifera) are the most significant perennial crop in arid regions of the Middle East and North Africa. Here, we present a comprehensive catalogue of approximately seven million single nucleotide polymorphisms in date palms based on whole genome re-sequencing of a collection of 62 cultivars. Population structure analysis indicates a major genetic divide between North Africa and the Middle East/South Asian date palms, with evidence of admixture in cultivars from Egypt and Sudan. Genome-wide scans for selection suggest at least 56 genomic regions associated with selective sweeps that may underlie geographic adaptation. We report candidate mutations for trait variation, including nonsense polymorphisms and presence/absence variation in gene content in pathways for key agronomic traits. We also identify a copia-like retrotransposon insertion polymorphism in the R2R3 myb-like orthologue of the oil palm virescens gene associated with fruit colour variation. This analysis documents patterns of post-domestication diversification and provides a genomic resource for this economically important perennial tree crop.

Comparative study of detection of bacteremia after different oral surgical procedures.

INTRODUCTION: The purpose of this study is to compare the efficiency of culture methods in detecting microorganisms appearing in the bloodstream after various oral surgical procedures and effect of preoperative antibiotics and antiseptic rinses on bacteremia. MATERIALS AND METHODS: The prevalence of bacteremia at various intervals of time (pre- and post-surgical) was studied in three different groups presenting with indications to surgical intervention. Two of the groups were given presurgical prophylaxis (systemic antibiotic and antiseptic rinse) while, in one group, no prophylactic measure was adopted. Blood samples were withdrawn from the subjects at specific pre- and post-surgical time intervals, and microbiological analysis was done. The findings were compared with a control group involving healthy individuals and a group involved in maxillofacial trauma where the breach of oral mucosal integrity was not surgically planned. RESULTS: The presence of bacteremia was not observed in all five groups at baseline while postsurgery (30 min, 60 min, and 90 min) in only three groups (preoperative antibiotic, without prophylaxis, and 1% Povidone iodine rinse). At all postsurgery periods (30 min, 60 min and 90 min), the presence of bacteremia was evident highest in without prophylaxis group followed by 1% Povidone iodine rinse and preoperative antibiotic the least.

DNA barcoding of Bemisia tabaci complex (Hemiptera: Aleyrodidae) reveals southerly expansion of the dominant whitefly species on cotton in Pakistan.

BACKGROUND: Although whiteflies (Bemisia tabaci complex) are an important pest of cotton in Pakistan, its taxonomic diversity is poorly understood. As DNA barcoding is an effective tool for resolving species complexes and analyzing species distributions, we used this approach to analyze genetic diversity in the B. tabaci complex and map the distribution of B. tabaci lineages in cotton growing areas of Pakistan. METHODS/PRINCIPAL FINDINGS: Sequence diversity in the DNA barcode region (mtCOI-5') was examined in 593 whiteflies from Pakistan to determine the number of whitefly species and their distributions in the cotton-growing areas of Punjab and Sindh provinces. These new records were integrated with another 173 barcode sequences for B. tabaci, most from India, to better understand regional whitefly diversity. The Barcode Index Number (BIN) System assigned the 766 sequences to 15 BINs, including nine from Pakistan. Representative specimens of each Pakistan BIN were analyzed for mtCOI-3' to allow their assignment to one of the putative species in the B. tabaci complex recognized on the basis of sequence variation in this gene region. This analysis revealed the presence of Asia II 1, Middle East-Asia Minor 1, Asia 1, Asia II 5, Asia II 7, and a new lineage "Pakistan". The first two taxa were found in both Punjab and Sindh, but Asia 1 was only detected in Sindh, while Asia II 5, Asia II 7 and "Pakistan" were only present in Punjab. The haplotype networks showed that most haplotypes of Asia II 1, a species implicated in transmission of the cotton leaf curl virus, occurred in both India and Pakistan. CONCLUSIONS: DNA barcodes successfully discriminated cryptic species in B. tabaci complex. The dominant haplotypes in the B. tabaci complex were shared by India and Pakistan. Asia II 1 was previously restricted to Punjab, but is now the dominant lineage in southern Sindh; its southward spread may have serious implications for cotton plantations in this region.

Pharmacokinetic interaction of ciprofloxacin with diclofenac: a single-dose, two-period crossover study in healthy adult volunteers.

BACKGROUND AND OBJECTIVE: Ciprofloxacin is a broad-spectrum, synthetic antibacterial used for the treatment of various bacterial infections. In multidrug therapy, ciprofloxacin is commonly prescribed with analgesics for the management of infection, pain and inflammation. The objective of this study was to evaluate the pharmacokinetic properties of ciprofloxacin tablets with concurrent administration of diclofenac tablets in healthy adult human volunteers. METHODS AND DESIGN: The disposition pharmacokinetics of a single oral dose of ciprofloxacin 500 mg alone and with co-administration of a diclofenac 50 mg tablet in 12 healthy male volunteers was investigated using a two-period, crossover design. The blood samples were collected at 0 (predose), 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 16 and 24 hours after administration of the drugs and the concentration of ciprofloxacin in serum was determined using reversed phase high-performance liquid chromatography. The pharmacokinetic parameters were calculated using a noncompartmental model and a two-compartment model. RESULTS: The maximum plasma concentration (C(max)) of ciprofloxacin increased from 2.48 +/- 0.33 microg/mL when administered alone to 3.91 +/- 0.8 microg/mL with co-administration of diclofenac. Time to reach C(max) (t(max)) with ciprofloxacin reduced from 2.02 +/- 0.3 hours when administered alone to 1.49 +/- 0.2 h with co-administration of diclofenac. Significant increases in ciprofloxacin area under the serum concentration-time curve (AUC) and elimination half-life, together with a significant decrease in total body clearance of ciprofloxacin, were observed with concurrent administration of diclofenac. CONCLUSION: Oral co-administration of ciprofloxacin tablets with diclofenac tablets increased ciprofloxacin AUC and C(max), and reduced ciprofloxacin t(max) and total body clearance.

Congenital giant melanocytic nevi.

Nevi are common skin tumors caused by abnormal overgrowth of cells from the epidermal and dermal layers of the skin. Most nevi are benign, but some pre-cancerous nevi must be monitored or removed. The giant congenital nevus is greater than 10 cm in size, pigmented and often hairy. Between 4% and 6% of these lesions will develop into a malignant melanoma. Since approximately 50% of the melanoma develop by the age of two, and 80% by the age of seven, early removal is recommended. The objective of this paper is to present a unique case of giant nevi and their surgical management.

Jarman index related to post-perinatal mortality.

Post-perinatal infant deaths in North and Southern Derbyshire District Health Authorities were calculated by electoral ward over a period of two years and related to Jarman scores of deprivation. The deaths were categorised into clinical-pathological groupings after full confidential enquiry. The post-perinatal death rates were significantly related to Jarman score of deprivation.

Differences in postperinatal infant deaths between north and southern Derbyshire.

In depth confidential inquiries on all post-perinatal infant deaths were carried out for two years in North and Southern Derbyshire District Health Authorities in order to explore why the postperinatal death rates and cot death rates have been consistently higher in Southern than in North Derbyshire. It was found that the death rates for probably inevitable deaths and for 'idiopathic' cot deaths were the same in both districts. The differences in the death rates lay in the group of partly explained cot deaths. Adverse social factors associated with each death were recorded and an estimate made of their relevance to the individual deaths. The group of deaths most amenable to intervention was those of category B (partly explained cot deaths). As judged by the Jarman index there are more areas of deprivation in Southern than in North Derbyshire.