A serological and molecular study on Francisella tularensis in rodents from Hamadan province, Western Iran.
Tularemia is a zoonotic disease, the most important hosts of which are rodents. Endemic regions and reservoirs of F. tularensis are not well-researched areas in Iran. The present study aimed to study F. tularensis infection in the rodent populations of western Iran. Samples were collected in different areas of Kabudar Ahang County in Hamadan province (west of Iran) from 2014 to 2017. Tularemia serological and molecular tests were conducted using the tube agglutination test and Real-time PCR method tracking the ISFtu2 gene. Positive serum samples were evaluated for cross-reactivity with brucellosis. A total of 433 rodents, collected from 33 localities, were included in the study. The most abundant species belonged to the Persian jird (Meriones persicus; 75.5%), and Libyan jird (Meriones libycus; 10.1%). Among the studied samples, three (0.74 %) were seropositive and five (1.15%) were PCR positive. Seropositive samples were two M. persicus and one M. libycus, and PCR positive rodents were four M. persicus and one M. vinogradovi. Tularemia seropositive samples showed no cross-reactivity with brucellosis. Given the presence of infection in rodents with tularemia agent in the studied area, it is crucial to elucidate the risks of rodent exposure to tularemia for physicians, health personnel and the general population.


Relationship between intracellular proton buffering capacity and intracellular pH.
In a recent publication the widely held view that the intracellular proton buffering power [defined as the amount of acid or base that has to be added to the cytosol to change the intracellular pH (pHi) by one pH unit] increases as the intracellular pH decreases, has been challenged, with the opposite relationship being proposed. In that publication, buffering was defined not in terms of pH change, but in terms of the change in proton concentration. The reason for this re-definition was the fear that the conventional analysis, using as it does a logarithmic function (pHi), could bias the outcome in favor of an increasing buffering power with decreasing pHi. The new system uses a "buffering co-efficient," defined as the number of protons necessary to be added to the cytosol to change the intracellular proton concentration by 1 mM. We report the use of both of these methods to analyze the relationship of pHi and buffering power, using human peripheral leucocytes loaded with the pH-sensitive fluorophore BCECF examined over a very wide range of pHi values (pHi 6.0 to 7.5). The most common method for pHi perturbation for the measurement of buffering is used, the rapid diffusion of ammonia across the cell membrane. In this study, analysis for both a bicarbonate-containing "open" system and for a Hepes-buffered "closed" system was performed.(ABSTRACT TRUNCATED AT 250 WORDS)


A comparison of the effects of lactose and of two chemically modified waxy maize starches on mineral metabolism in the rat.
Diets containing 30% by weight of waxy maize starch, lactose monohydrate, acetylated distarch phosphate (EEC No. 1414) or acetylated distarch adipate (EEC No. 1422) were fed to weanling female Specified Pathogen-Free Sprague-Dawley rats for 1 yr and to similar 9-month-old rats for 34 wk. Behaviour and general health were unaffected by the different diets and there were no diet-related differences in food consumption. AT the end of the experiment with 9-month-old rats the mean body weight of the animals receiving lactose was significantly lower than that of the controls receiving starch. The animals receiving the modified starches were slightly but not significantly heavier than the controls at the end of both experiments. The main treatment-related changes in rats on the three test diets were (1) caecal enlargement, (2) increased urinary excretion of calcium, (3) increased renal calcification as measured by chemical analysis of renal tissue obtained at autopsy and (4) increased medullary and pelvic nephrocalcinosis as assessed histopathologically. Acetylated distarch adipate had a slightly greater effect on the above parameters than acetylated distarch phosphate but both modified starches had less effect than lactose. The calcium content of the kidneys, as measured by chemical analysis or histopathology, increased with age, even in the animals receiving the control diet. This change may be due to excessively high concentrations of calcium and phosphorus in all the diets, including the control diet. Cortico-medullary mineral deposits were not a feature in these studies possibly because the diets were not deficient in magnesium. The importance of correct dietary formulation in long-term toxicity studies is emphasized.


Evaluation of transcriptional biomarkers using a high-resolution regression approach: Concentration-dependence of selected transcripts in copper-exposed freshwater mussels (Anodonta anatina).
We tested concentration-dependence of selected gene transcripts (cat, gst, hsp70, hsp90, mt and sod) for evaluation as biomarkers of chemical stress. Contrary to the common approach of factorial designs and few exposure concentrations, we used regression across a high-resolution concentration series. Specifically, freshwater mussels (Anodonta anatina) were acutely (96 h) exposed to Cu (13 nominal concentrations, measuring 0.13-1 600 µg/L), and transcripts were measured by RT-qPCR. In digestive glands, cat, hsp90 and mt decreased with water Cu (p < 0.05), but response magnitudes saturated at < 2-fold decreases. In gills, gst, hsp70, hsp90 and mt increased with water Cu (p < 0.05). While hsp70, hsp90 and mt exceeded 2-fold increases within the exposure range, high Cu concentrations were required (38-160 µg/L). Although gill responses were generally more robust compared to digestive glands, overall small response magnitudes and moderate sensitivity may set limit for potential application as general biomarkers of chemical stress.


Quantification of microbubble-induced sonothrombolysis in an ex vivo non-human primate model.
In vitro studies with ultrasound (US) and microbubbles (MB) have reported that sono-thrombolysis can be achieved at high peak rarefactional acoustic pressure amplitudes (PRAPAs) using 0.25 and 1.05 MHz US frequencies. The aim of the current study was to determine if these parameters work on an ex vivo physiological model of thrombosis. A thrombogenic device was placed in an ex vivo chronic arteriovenous shunt in juvenile baboons. Platelet accumulation was measured by dynamic imaging of the device and the 10 cm thrombus tail with 111 In-labeled platelets. After 15 minutes of thrombus formation, treatment with either low-dose recombinant tissue plasminogen activator (rtPA) or low-dose rtPA + MB+US was performed for 20 minutes. Four US settings at 0.25% duty cycle were used: 0.25 MHz at PRAPAs of 1.20 and 2.20 MPa, and 1.05 MHz at 1.75 and 4.75 MPa. Platelet accumulation was not inhibited by low-dose rtPA or MB with US alone. Platelet accumulation was significantly reduced with 0.25 MHz US at 2.20 PRAPA (P < .001) and with 1.05 MHz at 1.75 MPa and 4.75 MPa (P < .05) when used with MB and low-dose rtPA. Although this approach prevented platelet accumulation it did not cause thrombolysis on the device. rtPA + MB + US (0.25 and 1.05 MHz) resulted in inhibition of platelet accumulation on the thrombogenic device when moderately high PRAPAs (≥1.75 MPa) were used. These results taken in context with lytic effects of US on myocardial microthrombi and direct effect on myocardial blood flow and function provide direction for the use of therapeutic US in acute coronary syndromes.


The Efficacy and Safety of Iron Supplementation in Patients With Heart Failure and Iron Deficiency: A Systematic Review and Meta-analysis.
Iron deficiency (ID) is a common comorbidity in patients with heart failure (HF) and has been associated with increased mortality and hospitalizations. However, the benefit and safety of iron supplementation in treating HF and ID in randomized controlled trials (RCTs) are inconclusive. We therefore performed a meta-analysis to overcome this limitation. PubMed, the Cochrane Library, and ClinicalTrials.gov were systematically searched for eligible trials up to December 31, 2014. We also searched the references of all relevant studies and reviews for more trials. Only RCTs reporting the clinical impact of iron therapy in patients with HF with ID compared with no iron treatment were enrolled in our meta-analysis. Five clinical trials comprising a total of 907 patients were finally included. Compared with placebo or no treatment, additional iron therapy was associated with a significantly reduced rate of hospitalization for HF (odds ratio [OR], 0.28; 95% confidence interval [CI], 0.16-0.49), though all-cause mortality was not significantly different (OR, 0.81; 95% CI, 0.42-1.57). In 4 studies where these endpoints were combined, the incidence of hospitalization for HF and death was lowered in the iron supplementation group (OR, 0.47; 95% CI, 0.29-0.76). There was no increase in the risk of adverse events. Iron supplementation significantly reduced the risk of (a) hospitalization for HF and (b) the combined endpoint of hospitalization for HF and death, without increasing the risk of adverse events in patients with symptomatic systolic HF and ID. However, the current data are inadequate to make a clear determination upon mortality.


[Advantages of digital angiography in the study of vascular and parenchymal pathology in childhood].
The experience on 54 cases of vasculo-parenchimal pathology of paediatric age studied in our Institution in the last thirty (february '83-june '85) by venous or arterial DSA is reported. The advantages are analyzed provided by DSA in these patients, first represented by: highly satisfying and constant image quality obtained with general anaesthesia and controlled suspended breathing; minimal pharmacological and technical invasivity. A partial revision of diagnostic protocols, based on these considerations, may be proposed.


Antibody to fibroblast growth factor 23-peptide reduces excreta phosphorus of laying hens.
Novel strategies to minimize the excretion of phosphorus in swine and poultry are critical in minimizing environmental degradation. We have developed a synthetic peptide vaccine to produce autoantibodies to fibroblast growth factor 23 (FGF-23), a bone-derived hormone that blocks kidney phosphate resorption and indirectly reduces intestinal phosphate absorption. Single Comb White Leghorn laying hens, fed a standard diet (inorganic phosphorus, Pi = 0.4%), were immunized over the course of 4 weeks with either a FGF-23 peptide vaccine or adjuvant control (without FGF-23 peptide). At peak antibody titer to the peptide (week 5), 24-h excreta were collected and hens were blood sampled (represents 0.4% Pi treatment). Hens were then fed a 0.8% Pi diet and blood was sampled at 24 and 72 h and 24-h excreta were collected at 12 to 36 and 60 to 84 h (represents 0.8% Pi treatment). Increasing Pi from 0.4 to 0.8% increased (P < 0.05) percent excreta phosphorus, total 24-h phosphorus excretion, and plasma levels of FGF-23 and phosphate in either control or FGF-23 peptide vaccinated hens as early as the first sampling period. FGF-23 peptide vaccinated hens fed 0.4% Pi had reduced (P < 0.05) percent excreta phosphorus, total 24 h phosphorus excretion, and plasma levels of FGF-23 and iPTH, and increased (P < 0.05) plasma levels of phosphate and 1,25(OH)2D3 when compared to control vaccinated hens fed 0.4% Pi. In the first collection period post 0.8% Pi feeding, FGF-23 peptide vaccinated hens had reduced (P < 0.05) plasma levels of FGF-23 and iPTH, and increased (P < 0.05) plasma levels of phosphate and 1,25(OH)2D3, and tended to have reduced percent excreta phosphorus (P = 0.085) and total 24 h phosphorus excretion (P = 0.078) when compared to control vaccinated hens. Results during the second collection period post 0.8% Pi feeding were similar to that at the first collection period. These results are the first to show that the inhibition of FGF-23 action by a peptide vaccine (via neutralizing antibody) reduced phosphorus excretion. The approach presented provides new information on phosphorus metabolism in the laying hen.


Studies on the assembly of apo B-100-containing lipoproteins in HepG2 cells.
The relationship between apoB-100 and the membrane of the endoplasmic reticulum (ER) has been studied by a combination of pulse-chase methodology and subcellular fractionation. HepG2 cells were pulse-labeled with [35S]methionine for 3 min and chased with cold methionine for periods between 0 and 20 min. ApoB-100 and albumin, present in the membrane as well as in the luminal content of the ER vesicles, were isolated after each chase period. The results indicated that apoB-100 was cotranslationally bound to the membrane of the ER, and from this membrane-bound form, was transferred to the lumen after a delay of 10-15 min. Albumin was, as could be expected for a typical secretory protein, cotranslationally sequestered in the lumen of the ER. Apo-B-100-containing lipoproteins present in the microsomal lumen were analyzed by ultracentrifugation in a sucrose gradient. ApoB-100 occurred on rounded particles in three density regions: (i) d 1.1065-1.170 g/ml (Fraction I), (ii) d 1.011-1.045 g/ml (Fraction II), and (iii) d less than 1.011 g/ml (Fraction III). Fraction I, isolated from cells cultured in the absence of oleic acid, contained a homogenous population of particles with a mean diameter of approximately 200 A. Fraction I isolated from cells cultured in the presence of oleic acid was slightly more heterogeneous and had a mean diameter of approximately 250 A. Fractions II and III had mean diameters of 300 and 500 A, respectively. Cholesterol esters and triacylglycerol were the quantitatively dominating lipid constituents of all three fractions. Pulse-chase experiments indicated that Fraction I contained the newly assembled lipoproteins. With increasing chase time, the apoB-100 radioactivity was redistributed from Fraction I to Fractions II and III, indicating that Fraction I is converted into Fractions II and III during the intracellular transfer. Particles corresponding to Fractions II and III were by far the most abundant lipoproteins found in the medium. The results presented support the possibility of a sequential assembly of apoB-100-containing lipoproteins.


Reproductive performance of anovulatory anoestrous postpartum dairy cows following treatment with two progesterone and oestradiol benzoate-based protocols, with or without resynchrony.
(a) To compare the reproductive performance of anovulatory anoestrous (AA) postpartum dairy cows following treatment with 1 of 2 progesterone (P4) and oestradiol benzoate (ODB)-based treatment regimens; (b) To determine whether resynchronisation of cows initially treated for AA would improve reproductive performance and; (c) to determine whether cows not detected in oestrus but with a corpus luteum (CL) present (NDO/CL+), treated with P4 and ODB, would conceive earlier than untreated controls. Cows (n=1386) from 11 herds, that had not been detected in oestrus before the start of the seasonal mating period (PSM) and in which a CL was not detected were diagnosed AA, blocked by age (2 or >2 years old), then randomly assigned to be treated with an intravaginal P4-releasing device for either 6 (6-Day group) or 8 days (8-Day group). Cows in the 8-Day group were injected intramuscularly (IM) with 2 mg ODB at device insertion and all cows were injected with 1 mg ODB 24 h after device removal (Day 0). Cows detected in oestrus from Days 0-3 were subsequently assigned to be either resynchronised or left as untreated controls. Resynchronised cows had a used P4-releasing device reinserted on Day 14 for 8 days and were injected with 1 mg ODB at device reinsertion and again 24 h after device removal. NDO/CL+ cows were assigned to be either treated the same as the 8-Day group or left as untreated controls. All cows were inseminated on detection of oestrus and pregnancy tested approximately 10 weeks after the PSM and again 6-8 weeks after the end of the mating period. For AA cows, the 14-day submission rate was similar between 6-Day and 8-Day groups (p0.1). However, the pregnancy rate by Day 14 was higher for the 8-Day than the 6Day group (43.0% vs 35.2%; p=0.006). Resynchrony treatment decreased the 14-day pregnancy rate compared with untreated controls (35.2% vs 42.5%; p=0.026). The resynchrony treatment increased the submission rate between Days 14-28 for non-pregnant cows compared with untreated controls (80.6% vs 57.4%; p=0.049). However, conception rate to resynchronised heats was lower than for cows that returned to oestrus naturally (56.6% vs 67.9%; p=0.025). Neither initial treatment type nor resynchrony treatment increased the 28-day pregnancy rate (p>0.1). There were no differences between treatment groups in the final non-pregnant rate (4.5% vs 4.6%; p>0.1). Treated NDO/CL+ cows had a higher 14-day submission rate (88.1% vs 49.4%; p>0.001), higher 14-day and 28-day pregnancy rates (42.9% vs 20.7%, p>0.001 and; 56.0% vs 42.5%, p=0.094, respectively) and conceived earlier (21 vs 36 days from PSM to median day of conception; p>0.05), than untreated NDO/CL+ cows. The 8-Day, ODB-P4-ODB treatment regimen resulted in a higher pregnancy rate by 14 days but not 28 days than the 6-Day, P4-ODB treatment. The resynchrony treatment increased the proportion of non-pregnant cows inseminated on days 14-28, but did not increase the 28-day pregnancy rate or final pregnancy rate. Treatment of NDO/CL+ cows with the 8-Day, ODB-P4-ODB treatment improved reproductive performance compared with no treatment.


Treatment of Chronic Hepatitis C in Adolescent Patients With Positive HBsAg or With Occult Hepatitis B: Is the Risk of Hepatitis B Reactivation Significant?
Reactivation of hepatitis B virus (HBV) infection in patients treated for chronic hepatitis C (HCV) with direct-acting antiviral agents has emerged recently as an important safety issue; however, it has not been adequately studied in pediatric age groups. We aimed to evaluate this risk in adolescent patients infected with chronic HCV and positive for HBsAg and HBcAbs. One hundred and fifteen adolescent patients from 12 to 17 years of age, infected with chronic HCV and positive for HBcAbs with or without HBsAg were included in this study. All patients were treated with 1 tablet daily of the fixed-dose combination sofosbuvir/ledipasvir for 12 weeks. Patients were closely monitored throughout the study for virus load, liver functions, and other safety and efficacy outcome measures. The sustained virologic response 12 (SVR12) rates were 96.7% (95% confidence interval: 88.6-99.1%) in HBsAg positive group and 98.2% (95% confidence interval: 90.4-99.7%) in HBsAg negative with HBcAbs positive group. Throughout the treatment period and the 12 weeks follow-up after treatment, there has been no single case in both HBsAg negative or positive that showed any manifestation of reactivation of hepatitis B, detected levels of HBV-DNA, or deterioration of liver functions. No HBV reactivation was observed in adolescents treated for chronic HCV with direct-acting antiviral agents in our study, in both HBsAg positive or occult hepatitis B. Although results are reassuring, we still recommend close monitoring of liver functions to not miss even rare cases of such a potentially serious condition.


Insulin-like growth factor-1 receptor polymorphism and ischemic stroke: a case-control study in Chinese population.
Low levels of insulin-like growth factor 1 (IGF-1) are associated with atherosclerosis, and insulin-like growth factor-1 receptor (IGF-1R) polymorphisms can change plasma levels of IGF-1 and may alter the function of the receptor. Whether there is any association of genetic variation in IGF-1R gene with ischemic stroke (IS) is presently unknown. A 1:1 case-control study was conducted. The G --> A polymorphism of IGF-1R gene (rs2229765) were analyzed by TaqMan SNP genotyping technique in Chinese patients with IS (n = 309) and old subjects without IS (n = 309). The frequency of A allele in the patients and controls was 45.79% and 39.64%, respectively. The AA genotype distribution of IGF-1R gene was significantly higher in the patients (27.51%) than controls (18.23%; P = 0.022). Conditional logistic regression revealed that the AA genotype of IGF-1R was associated with IS (OR = 1.641, P = 0.022). After adjustment for smoking, alcohol drinking, history of hypertension, and body mass index, IGF-1R AA genotype was still significantly associated with an increased risk of IS (OR = 1.787, P = 0.029), compared with IGF-1R GG. The G --> A polymorphism in IGF-1R gene may affect the susceptibility to IS in Chinese population.


Axitinib pharmacologic therapeutic monitoring reveals severe under-exposure despite titration in patients with metastatic renal cell carcinoma.
Introduction New therapeutic strategies combining axitinib and immune checkpoint blockers are ongoing in metastatic renal cell carcinoma (mRCC). These strategies do not consider the pharmacokinetic variability of axitinib. We aimed to describe the risk of axitinib under-exposure using routine pharmacologic therapeutic monitoring (PTM). Methods We analyzed axitinib dosage in nine patients with mRCC. Routine axitinib concentration measurements were centralized at Henri Mondor University Hospital (Créteil, France) using a validated method. The primary objective was to describe the evolution of Cmax dosages (1 to 6 h after oral intake) during routine axitinib titration. Results Nine patients with available Cmax axitinib dosages were included. Four out of the nine patients had axitinib titration and Cmax dosages were performed before and after titration. All but one corrected their plasma axitinib exposure after titration, suggesting of a titration success. The last patient was monitored in the Henri Mondor Hospital routine PTM program and a pharmacokinetic profiling was performed after controlled oral intake. Results suggested a poor axitinib absorption. This patient experienced early tumor progression as best response. Conclusion We report a patient with significant axitinib under-exposure, possibly due to a poor absorption. PTM should be evaluated and considered in drug developments evaluating combination therapies based on axitinib.


Sulforaphane prevents and reverses allergic airways disease in mice via anti-inflammatory, antioxidant, and epigenetic mechanisms.
Sulforaphane has been investigated in human pathologies and preclinical models of airway diseases. To provide further mechanistic insights, we explored L-sulforaphane (LSF) in the ovalbumin (OVA)-induced chronic allergic airways murine model, with key hallmarks of asthma. Histological analysis indicated that LSF prevented or reversed OVA-induced epithelial thickening, collagen deposition, goblet cell metaplasia, and inflammation. Well-known antioxidant and anti-inflammatory mechanisms contribute to the beneficial effects of LSF. Fourier transform infrared microspectroscopy revealed altered composition of macromolecules, following OVA sensitization, which were restored by LSF. RNA sequencing in human peripheral blood mononuclear cells highlighted the anti-inflammatory signature of LSF. Findings indicated that LSF may alter gene expression via an epigenetic mechanism which involves regulation of protein acetylation status. LSF resulted in histone and α-tubulin hyperacetylation in vivo, and cellular and enzymatic assays indicated decreased expression and modest histone deacetylase (HDAC) inhibition activity, in comparison with the well-known pan-HDAC inhibitor suberoylanilide hydroxamic acid (SAHA). Molecular modeling confirmed interaction of LSF and LSF metabolites with the catalytic domain of metal-dependent HDAC enzymes. More generally, this study confirmed known mechanisms and identified potential epigenetic pathways accounting for the protective effects and provide support for the potential clinical utility of LSF in allergic airways disease.


Perinatal and early life risk factors for childhood brain tumors: Is instrument-assisted delivery associated with higher risk?
The childhood peak of brain tumors suggests that early-life exposures might have a role in their etiology. Hence, we examined in the Greek National Registry for Childhood Hematological Malignancies and Solid tumors (NARECHEM-ST) whether perinatal and early-life risk factors influence the risk of childhood brain tumors. In a nationwide case-control study, we included 203 cases (0-14 years) with a diagnosis of brain tumor in NARECHEM-ST (2010-2016) and 406 age-, sex-, and center-matched hospital controls. Information was collected via interviews with the guardians and we analyzed the variables of interest in multivariable conditional logistic regression models. Instrument-assisted delivery was associated with higher (OR: 7.82, 95%CI: 2.18-28.03), whereas caesarean delivery with lower (OR: 0.67, 95%CI: 0.45-0.99) risk of childhood brain tumors, as compared to spontaneous vaginal delivery. Maternal alcohol consumption during pregnancy (OR: 2.35, 95%CI: 1.45-3.81) and history of living in a farm (OR: 4.98, 2.40-10.32) increased the odds of childhood brain tumors. Conversely, higher birth order was associated with lower risk (OR for 2nd vs. 1st child: 0.60, 95%CI: 0.40-0.89 and OR for 3rd vs. 1st: 0.34, 95%CI: 0.18-0.63). Birth weight, gestational age, parental age, history of infertility, smoking during pregnancy, allergic diseases, and maternal diseases during pregnancy showed no significant associations. Perinatal and early-life risk factors, and specifically indicators of brain trauma, exposure to toxic agents and immune system maturation, might be involved in the pathogenesis of childhood brain tumors. Larger studies should aim to replicate our findings and examine associations with tumor subtypes.


[Etanercept].
Etanercept (ETN) is a fusion protein of the receptor (CD120b) for tumor necrosis factor (TNF) and the Fc portion of IgG1. A phase III trail and the JESMR study, both performed in Japan, implicated the importance of concomitant methotrexate (MTX) use in the treatment of rheumatoid arthritis (RA). Further evidences, including the TEAR study and the PRESERVE study, supported the recent treatment strategy for RA, being divided into at least two phases: remission induction phase(step-up or initial combination) and its maintenance phase (step-down). Thus, optimization of RA treatment chiefly based on MTX and anti-TNF biological agents is in progress.


Adenovirus-mediated expression of p33(ING1b) induces apoptosis and inhibits proliferation in gastric adenocarcinoma cells in vitro.
Inhibitor of growth 1b (ING1b) is considered to be a class II tumor suppressor gene. Although reduced expression of p33(ING1b) has been reported in many human malignancies, including gastric cancers, the effect of p33(ING1b) on gastric cancer cells has yet to be investigated. Expression of p33(ING1b) in gastric adenocarcinoma tissues and their adjacent non-malignant gastric mucosa, as well as in gastric adenocarcinoma cell lines and normal gastric epithelial cells, was detected by using Western blotting. Recombinant adenoviruses were prepared to mediate the ectopic expression of p33(ING1b) (Ad-ING1b) and green fluorescent protein (GFP)(Ad-GFP) in the gastric adenocarcinoma cell lines, SGC-7901, MKN28, and MKN45 and the normal gastric epithelial cell line GES-1. Alterations in the proliferation and apoptosis of the cells after adenoviral infection were determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively, and cell cycle distribution was analyzed in a fluorescence-activated cell sorter. Western blotting confirmed the reduced expression of p33(ING1b) in gastric adenocarcinoma tissues and gastric adenocarcinoma cell lines. The ectopic expression of p33(ING1b) mediated by Ad-ING1b resulted in decreased growth, increased apoptosis, and cell cycle arrest at the G1 phase in both benign and malignant gastric epithelial cells regardless of their p53 status. Addition of a p53 inhibitor, pifithrin-α, did not abolish the pro-apoptotic and cell cycle-arresting effects of p33(ING1b) in p53 wild-type cells. Down-regulation of p33(ING1b) might play an important role in the development of gastric adenocarcinoma. Targeted local expression of p33(ING1b) may offer a promising alternative therapeutic measure for gastric cancer.


Reliability and accuracy in radiographic measurements of musculoskeletal tumors.
Tumor size and growth are important parameters when evaluating bone and soft tissue neoplasms. There are no reports comparing the intra- and interobserver reliability among physicians in their evaluation of musculoskeletal (MSK) tumor imaging. This study investigates the accuracy and precision of measurements made by orthopedic and radiology physicians in different stages of training. Blinded magnetic resonance imaging (MRI) scans from six patients, three soft tissue, and three bone tumors were selected: each case included an "old" and "new" scan that was performed at least 3 months apart. Fourteen participants were selected, representing varying levels of education and experience, including two of each of the following: medical students, orthopedic and radiology residents, oncology and nononcologic orthopedic attendings, and MSK and non-MSK radiology attendings. Participants compared the old and new studies, recording tumor size in the transverse, cranial-caudal, and anterior-posterior dimensions, and determined if the tumor was stable or unstable. The MRI's official report served as the "gold standard." Average intraobserver variability (|Trial 1 - Trial 2|/[(Trial 1 + Trial 2)/2])) in size measurements was 11.08% (0.00%-68.62%). The lowest variability was recorded by the MSK radiologist 1 (6.16%), and the greatest variability by Orthopedic Surgery Resident 1 (16.70%). Participants correctly determined stability 82% of the time (71%-100%). Only MSK radiologists correctly determined stability in over 90% of cases. There is considerable variability and inaccuracy in MRI-based measurements of MSK tumors. These findings motivate opportunities for improving MSK imaging education of radiology and orthopedic residents. Physicians ordering MRI scans should evaluate them themselves, instead of relying on the radiology report alone, to inform clinical decision-making.


Tick infestation on caimans: a casual tick-host association in the Atlantic rainforest biome?
Ticks are parasites of birds, mammals, amphibians, and reptiles, but information about tick communities that parasitize reptiles in the Neotropical region is still fragmentary. In the present study, we assessed the presence of ticks on broad-snouted caimans (Caiman latirostris) and Cuvier's dwarf caimans (Paleosuchus palpebrosus) trapped in the Atlantic rainforest biome in Pernambuco state, north-eastern Brazil, to determine which tick species feed on these animals and how frequent or rare this parasite-wildlife association is. We also report an occasional finding of Amblyomma rotundatum on a smooth-fronted caiman (Paleosuchus trigonatus) in the Amazon biome in Pará state, northern Brazil. Out of 490 animals trapped in the Atlantic rainforest biome, four (0.82%) broad-snouted caimans were infested by ticks. Ticks belonged to two Amblyomma species: A. rotundatum (three females) and A. fuscum (one male). Our findings indicate that ticks are infrequent parasites of caimans in the Atlantic rainforest biome. Tick infestation on broad-snouted caimans is probably of minor clinical significance and probably a casual finding due to habitat sharing with the common tick hosts.


Softgel capsule technology as an enhancer device for the absorption of natural principles in humans. A bioavailability cross-over randomised study on silybin.
In order to evaluate if a patented soft gelatine capsule could improve the bioavailability of silybin (CAS 22888-70-6) in comparison to a hard shell capsule, an open, single dose, two-way, balanced cross-over study, was performed. The study was conducted on 12 healthy subjects (6 M and 6 F). 80 mg of silybin in a 1:2 complex with phosphatidylcholine was administered. Blood was sampled from the subjects in two occasions at the following times after drug administrations: 0 (sample before dosing), 1, 2, 3, 4, 6 and 8 h. The pharmacokinetic parameters calculated from the results of the plasma analyses demonstrated that the mean values of both Cmax and AUC0-1 were increased when the patented soft gelatine capsule formulations were administered (i.e. Cmax more than 3-fold and AUC0-1 more than 2-fold).


Psychometric testing of the Chinese Mandarin version of the Medical Outcomes Study Social Support Survey in patients with coronary heart disease in mainland China.
To evaluate the psychometric properties of the Chinese Mandarin version of the Medical Outcomes Study Social Support Survey (MOS-SSS-CM) in patients with coronary heart disease (CHD) in mainland China. Measurement performance of the MOS-SSS-CM was tested on a convenience sample of 200 Chinese patients with CHD in a University-affiliated hospital in Xi'an, P. R. China. To establish reliability of the instrument, 40 patients were retested 2 weeks later through telephone interview. The traditional Chinese version of the MOS-SSS was transformed into a simplified Chinese Mandarin version and administered to participants, together with Chinese Mandarin versions of the Short Form-36 item Health Survey (SF-36) and the Hospital Anxiety and Depression Scale (HADS). The MOS-SSS-CM had acceptable internal consistency with Cronbach α coefficients of 0.91 for the overall scale and 0.71-0.84 for the four subscales. The high correlation (r = 0.56-0.87) between items and the remainder of the scale provides further evidence of internal consistency. The test-retest reliability was generally acceptable with intraclass correlation coefficients of 0.89 for the overall scale and 0.74-0.88 for the four subscales. There was acceptable concurrent validity with moderate significant correlations (r ≥ 0.3, p < 0.01) between the MOS-SSS-CM and the Chinese Mandarin version of the HADS. Confirmatory factor analysis supported a four-factor structure of the MOS-SSS-CM measuring the self-perceived adequacy of functional support of Chinese patients with CHD. The MOS-SSS-CM is a valid and reliable measure for Chinese Mandarin-speaking patients with CHD.


A CASE OF RAPIDLY PROGRESSIVE RETINOPATHY ASSOCIATED WITH PEMBROLIZUMAB IMMUNOTHERAPY FOR METASTATIC UROTHELIAL CARCINOMA.
To describe a novel case of bilateral rapidly progressive retinopathy after immunotherapy with pembrolizumab for metastatic urothelial carcinoma. Case report. A 64-year-old man undergoing pembrolizumab immunotherapy was referred to our hospital because of bilateral acute vision loss. His best-corrected visual acuity was 20/30 in the right eye and 20/320 in the left eye, and a visual field test revealed central and paracentral scotomas in the right eye and central scotoma in the left eye. We suspected immune-related retinopathy based on the progressive photoreceptor damage with abnormal electroretinogram findings, absence of overt intraocular inflammation, and presence of malignancy. Cessation of pembrolizumab and steroid pulse therapy followed by decreasing oral prednisolone improved visual function and photoreceptor damage, although there was recurrence after pembrolizumab was restarted. We reported a case of rapidly progressive retinopathy that may have been triggered by pembrolizumab immunotherapy for metastatic urothelial carcinoma. High-dose corticosteroid pulse therapy was effective in reversing photoreceptor damage.


Aortic valve calcification in hypertensive patients: prevalence, risk factors and association with transvalvular flow velocity.
The important role of the "nonobstructive" aortic valve calcification (AVC) in cardiovascular morbidity and mortality has recently been emphasized. The present work had two goals: (1) to analyze the prevalence and factors determining the extent of AVC in hypertensive patients; (2) to investigate a possible association between the extent of AVC and flow velocity across the aortic valve. This was a prospective study performed in a university hospital. The sample included 263 consecutive patients (139 men and 124 women), mean age 65+/-6, who underwent echo-Doppler. The upper quartile of peak flow velocity across the aortic valve (>130 mm/s in our population) was defined as augmented flow velocity. There were 31 (12%) patients in the advanced AVC group and 122 (46%) without any calcified deposits (the no AVC group). The remaining 110 (42%), who did not meet criteria for advanced AVC, comprised the trivial AVC group. Peak flow velocity was significantly higher in patients with advanced vs. trivial AVC and no AVC groups: 135+/-45, 116+/-23 and 113+/-23 cm/s, respectively; p=0.0002. Prevalence of augmented transvalvular aortic flow was significantly higher (p=0.01) among patients with advanced AVC (41.9%) vs. trivial (20.9%) and no AVC (20.5%). Multivariate analyses identified age as the only independent variable associated with advanced AVC [OR 1.6 (CI 1.2-2.3), 5 years increment]. Advanced AVC and female gender were identified as independent variables for augmented transvalvular aortic flow with OR 2.9 (CI 1.3-6.4) and 2.5 (CI 1.4-4.6), respectively. Prevalence of AVC among hypertensive patients is high and clearly age-related. Female gender and advanced (but not trivial) AVC are associated with augmented aortic transvalvular peak flow. Our results stress the role of protruding calcium deposits in augmentation of rest flow velocity across the aortic valve regardless of visible restriction of leaflet opening.


Isolation of partial toxR gene of Vibrio harveyi and design of toxR-targeted PCR primers for species detection.
To differentiate Vibrio harveyi from closely related Vibrio species by toxR sequence analysis and design primers for the specific detection of the shellfish pathogen. The partial toxR homologue from the shellfish pathogen V. harveyi was isolated by PCR using degenerate primers. The 578-bp toxR fragment from V. harveyi, that exhibited highest homology with partial toxR of V. parahaemolyticus (68%), is predicted to encode for a polypeptide with 192 amino acid residues. Alignment of the V. harveyi toxR nucleotide and deduced amino acid sequence with those from other Vibrio species revealed the presence of the fairly characteristic conserved transcription activation and transmembrane domain as well as the divergent membrane tether region that may be targeted for the development of species-specific oligonucleotide primers. Consequently, PCR primers that could amplify a 390-bp gene fragment in V. harveyi were designed by targeting portions of the V. harveyi toxR that display variability with toxR sequences from other Vibrio species. The 390-bp-amplicon was detected in all V. harveyi strains examined except in the nontarget bacteria and unexpectedly, in two shrimp-derived strains (VIB 391 and STD 3-101) from Thailand and Ecuador. Results show that strains exhibiting the 390-bp amplicon mostly belong to the same cluster based on previous amplified fragment length polymorphism data while strains which were previously unclustered or unclassified did not display the 390-bp PCR product. The toxR sequence variation could differentiate V. harveyi from closely related Vibrio species. A PCR protocol amplifying a 390-bp fragment of the V. harveyi toxR was established and could be useful in the specific and rapid detection of the species. The molecular approaches reported in this study could facilitate the early diagnosis and surveillance of luminous vibriosis in hatchery-reared fish and shellfish species through rapid identification and specific detection of causal agent.


Allele frequency covariance among cohorts and its use in estimating effective size of age-structured populations.
A general expression for the covariance of allele frequencies among cohorts in age-structured populations is derived. The expression is used to extend the so-called temporal method for estimating effective population size from allele frequency shifts among samples from cohorts born any number of years apart. Computer simulations are used to check on the accuracy and precision of the method, and an application to coastal Atlantic cod is presented.


The significance of the metabolism of the neurohormone melatonin: antioxidative protection and formation of bioactive substances.
Recent findings suggest that the ability of melatonin to enter all body tissues and to be metabolized, enzymatically or nonenzymatically, in any of them results in a spectrum of effects, which exceed substantially those transduced by membrane receptors. These actions comprise the formation of various bioactive compounds such as N-acetylserotonin, 5-methoxytryptamine, N,N-dimethyl-5-methoxytryptamine, 5-methoxytryptophol, cyclic 2-hydroxymelatonin, pinoline, and 5-methoxylated kynuramines. Apart from enzymatic metabolism, nonenzymatic reactions with free radicals, in particular the superoxide anion and the hydroxyl radical, represent a new and significant aspect of melatonin's biological role. Melatonin represents the most potent physiological scavenger of hydroxyl radicals found to date, and recent findings suggest an essential role of this indoleamine for protection from hydroxyl radical-induced carcinogenesis and neurodegeneration.


[Peripheral mechanisms of the responses of small intestine capacitance vessels to noradrenaline].
Noradrenalin was shown to elicit either constrictory or dilatory responses depending on the initial level of venous pressure (10-15 mm Hg or 0 and 25 mm Hg) in the capacitance vessels of the cat small intestine. Blockade of beta-adrenoreceptors with propranolol has no effect on the capacitance vessels responses. Blockade of alpha-adrenoreceptors with dihydroergotoxin makes noradrenaline to elicit dilatory responses only, their magnitude depending on the initial level of venous pressure: more obvious responses at 10-15 mm Hg, less obvious those - at 0 and 25 mm Hg.


"Triple-H" therapy for cerebral vasospasm following subarachnoid hemorrhage.
The combination of induced hypertension, hypervolemia, and hemodilution (triple-H therapy) is often utilized to prevent and treat cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH). Although this paradigm has gained widespread acceptance over the past 20 years, the efficacy of triple-H therapy and its precise role in the management of the acute phase of SAH remains uncertain. In addition, triple-H therapy may carry significant medical morbidity, including pulmonary edema, myocardial ischemia, hyponatremia, renal medullary washout, indwelling catheter-related complications, cerebral hemorrhage, and cerebral edema. This review examines the evidence underlying the implementation of triple-H therapy, and makes practical recommendations for the use of this therapy in patients with aneurysmal SAH.


High level expression, efficient purification and bioactivity assay of recombinant human platelet-derived growth factor AA dimer (PDGF-AA) from methylotrophic yeast Pichia pastoris.
Platelet-derived growth factors (PDGFs) are important biochemical mediators regulating many physiological and pathophysiological processes, including promotion of the chemotactic recruitment and proliferation of cells involved in wound repair. Previously, homodimers of rhPDGF-AA protein were purified from Escherichia coli. However, eukaryotic proteins often contain posttranslational modifications, such as glycosylation, that are required for biological functions. In this study, an efficient method was established to purify a glycosylated rhPDGF-AA dimer from P. pastoris culture media by one step CM Sepharose ion exchange chromatography yielding about 20mg/L of over 95% highly purified rhPDGF-AA. Mass spectrometry analysis of the purified rhPDGF-AA displayed a molecular weight (MW) of 27,825.513Da, composed of a subunit with MW of 15,042.945Da and a subunit with MW of 12,904.374Da. The size difference is accounted for by differential glycosylation of the monomers. Biological activity of the rhPDGF-AA was confirmed by its ability to induce NIH/3T3 cells proliferation. The experimental procedure we have developed facilitates production of an active glycosylated rhPDGF-AA in large amounts for further research and drug development.


Syngeneic lymph-node-targeting model of green fluorescent protein-expressing Lewis lung carcinoma.
The Lewis lung tumor has been extensively studied in both syngeneic and allogeneic mouse models. However, its metastatic potential and mechanism are poorly understood. The aim of the present study was to develop a highly metastatic lymph-node targeting, imageable model of the Lewis lung carcinoma in a syngeneic host. We report here a syngeneic model of the Lewis lung carcinoma in which the carcinoma cells are labeled with green fluorescent protein (GFP). The tumor cells were transplanted in the dorsal side of the ear of C57-B16 mice in order to give the tumor cells access to the lymphatic system. This model of the Lewis lung carcinoma extensively metastasized to numerous lymph nodes throughout the body of the animal as well as visceral organs, as visualized by fluorescence microscopy using the bright GFP signal. Twenty-one different metastatic sites, including lymph nodes throughout the body, were identified among the cohort of transplanted animals. The data demonstrate a predilection of the Lewis lung carcinoma for lymphatic pathways for metastasis throughout the animal body. The concomitant macrometastases to the visceral organs observed in this model may be remetastasis from the lymph nodes. This model of the Lewis lung carcinoma should be very useful in defining cellular trafficking and targeting mechanisms of metastasis, in particular those involving lymphatic pathways.


Standardized echographic-histopathologic correlations in liposarcoma.
A 57-year-old woman had metastatic liposarcoma of the left orbit that underwent transformation. The primary focus in the abdomen, resected in 1975, was a well-differentiated liposarcoma. Later metastases to the neck and orbit were poorly differentiated, pleomorphic, and highly anaplastic liposarcomas. The ultrasonographic and histopathologic correlations of the different stages of the tumor during the disease process were analyzed and compared. This patient's disease had echopathologic similarities to other orbital sarcomas and adult cavernous hemangiomas.


Improving access to healthcare for women in Tanzania by addressing socioeconomic determinants and health insurance: a population-based cross-sectional survey.
This study was performed to explore the factors associated with accumulation of multiple problems in accessing healthcare among women in Tanzania as an example of a low-income country. Population-based cross-sectional survey. Nationwide representative data for women of reproductive age obtained from the 2015-2016 Tanzania Demographic and Health Survey were analysed. A composite variable, 'problems in accessing healthcare', with five (1-5) categories was created based on the number of problems reported: obtaining permission to go to the doctor, obtaining money to pay for advice or treatment, distance to a health facility and not wanting to go alone. Respondents who reported fewer or more problems placed in lower and higher categories, respectively. A total of 13 266 women aged 15-49 years, with a median age (IQR) of 27 (20-36) years were interviewed and included in the analysis. About two-thirds (65.53%) of the respondents reported at least one of the four major problems in accessing healthcare. Furthermore, after controlling for other variables included in the final model, women without any type of health insurance, those belonging to the poorest class according to the wealth index, those who had not attended any type of formal education, those who were not employed for cash, each year of increasing age and those who were divorced, separated or widowed were associated with greater problems in accessing healthcare. This study indicated the additive effects of barriers to healthcare in low-income countries such as Tanzania. Based on these results, improving uptake of health insurance and addressing social determinants of health are the first steps towards reducing women's problems associated with accessing healthcare.


Comparison of anti-malarial drugs efficacy in the treatment of uncomplicated malaria in African children and adults using network meta-analysis.
Artemisinin-based combination therapy (ACT) and novel drug combinations are available and used in African countries to treat uncomplicated malaria. Network meta-analysis methods are rarely and poorly applied for the comparison of their efficacies. This method was applied on a set of randomized controlled trials to illustrate its usefulness. A literature review available in Pubmed was conducted in July 2016. Eligible studies, conducted in sub-Saharan Africa, published between 2002 and 2016, focused on randomized controlled trials of at least two artemisinin-based combinations to treat uncomplicated malaria in children and adults. Agglomerate data were: the number of PCR-corrected adequate clinical and parasitological response (ACPR) on day 28, used as the primary enDHAPoint in all interventions, the number of participants and the list of treatments. A Bayesian random effect meta-analysis using a binary outcome was the method to compare the efficacy. Ranking measure was used to obtain a hierarchy of the competing interventions. In total, 76 articles were included; 13 treatment regimens were involved and tested in 36,001 patients. Using artemether-lumefantrine (AL) as the common comparator for the entire network, 12 relative treatment effects were estimated and indirect comparisons were obtained. Dihydroartemisinin-piperaquine (DHAP) was shown to be more effective than AL (odds ratio [OR] = 1.92; 95% CI 1.30-2.82; 19,163 patients), ASAQ (OR = 1.70; 95% CI 1.10-2.64; 14,433 patients), and amodiaquine-sulfadoxine-pyrimethamine (AQSP): OR = 2.20; 95% CI 1.21-3.96; 8863 patients. Artesunate-amodiaquine (ASAQ) was comparable to AL (OR = 1.11; 95% CI 0.84-1.45; 21,235 patients). No significant difference was found between artesunate and mefloquine (ASMQ) and AL (OR = 1.20; 95% CI = 0.52-2.8; 13,824 participants). According to treatment ranking, among the WHO-recommended ACT medicines, DHAP was shown to be the most efficacious. Based on the available evidence, this study demonstrated the superiority of DHAP among currently recommended artemisinin-based combinations. The application of the methods described here may be helpful to gain better understanding of treatment efficacy and improve future decisions. However, more data are needed to allow robust conclusions about the results in comparison with novel drugs. Further surveillance of the efficacy of anti-malarial drugs and clinical trials are needed to closely follow the evolution of the epidemiology of drug-resistant malaria in Africa.


A phenylalanine in peptide substrates provides for selectivity between cGMP- and cAMP-dependent protein kinases.
Bovine lung cGMP-binding cGMP-specific phosphodiesterase (cG-BPDE) is a potent and relatively specific substrate for cGMP-dependent protein kinase (cGK) as compared to cAMP-dependent protein kinase (cAK) (Thomas, M. K., Francis, S. H., and Corbin, J. D. (1990) J. Biol. Chem. 265, 14971-14978). A synthetic peptide, RKISASEFDRPLR (BPDEtide), was synthesized corresponding to the sequence surrounding the phosphorylation site in cG-BPDE. BPDEtide retained the cGK/cAK kinase specificity demonstrated by native cG-BPDE: the apparent Km of BPDEtide for cGK was 5-fold lower than that for cAK (Km = 68 and 320 microM, respectively). Vmax values were 11 mumol/min/mg for cGK and 3.2 mumol/min/mg for cAK. The peptide was not phosphorylated to a measurable extent by protein kinase C or by calcium/calmodulin-dependent protein kinase II. Thus, the primary amino acid sequence of the peptide substrate was sufficient to confer kinase specificity. Studies in crude tissue extracts indicated that BPDEtide was the most selective peptide substrate documented for measuring cGK activity. Peptide analogs of BPDEtide were synthesized to determine the contribution of specific residues to cGK or cAK substrate specificity. Substitution of a Lys for the amino-terminal Arg did not reduce cGK/cAK specificity; neither did the exchange of an Ala for the non-phosphorylated Ser nor the removal of the 3 carboxyl-terminal residues. A truncated BPDEtide (RKISASE) served equally well as substrate (Km approximately 90 microM) for both kinases. However, restoration of the Phe, to yield RKISASEF, reproduced the original cGK/cAK specificity for BPDEtide (Km = 120 and 480 microM, respectively), primarily by decreasing the affinity of cAK. Addition of a carboxyl-terminal Phe to the peptide RKRSRAE (derived from the sequence of the cGK phosphorylation site in histone H2B) or to the peptide LRRASLG (derived from the sequence of the cAK phosphorylation site in pyruvate kinase) also improved the cGK/cAK specificity by decreasing the affinity of cAK. These data suggested that the Phe in each substrate tested is a negative determinant for cAK.


Surgical aspects and early morbidity of patients undergoing open recurrent inguinal hernia repair.
This study aims to characterize the patterns of recurrence associated with specific types of primary inguinal hernia repair techniques used for and their respective correlations with early morbidity, in patients undergoing open repair for their first hernia recurrence. After ethics approval was obtained, a retrospective review of charts from patients who underwent open surgery for repair of a first recurrence after a previous inguinal hernia repair during 2013-2017 was completed. Statistical analyses were performed and p-values < .05 are reported as statistically significant. 1393 patients underwent 1453 surgeries for recurrent inguinal hernias at this institution. Operations for recurrence were longer (61.9 ± 21.1 vs. 49.3 ± 11.9; p < .001), required more frequent intra-operative surgical consultation (1% vs. 0.2%; p < .001) and had a higher incidence of surgical-site infections (0.8% vs. 0.4%; p = .03) than primary inguinal hernia repairs. When comparing the patterns of recurrence among different techniques of primary repairs, patients undergoing laparoscopic hernia repair presented with a higher incidence of indirect recurrences. Reoperations after a Shouldice repair and open mesh repair represented markers for higher surgical difficulty in the recurrent operation (longer operative time, higher identification of heavy scarring, less nerve identification, and higher frequency of intra-operative consultation), but not higher rates of complications when compared with other techniques. Open reoperations for inguinal hernia first recurrences are more complex, with noticeable differences according to the index operation, and associated with higher morbidity when compared with primary hernia repairs. This complexity varies according to the type of primary surgery, with a previous Shouldice repair and open hernia repair with mesh presenting higher surgical difficulties although this did not translate to higher incidence of early complications. This information may allow adequate allocation of surgeons with an expertise in recurrent hernias and choice of recurrent repair method (laparoscopic or open) based on the primary surgery.


Single midline thalamic neurons projecting to both the ventral striatum and the prefrontal cortex in the rat.
The midline thalamic nuclei have been known to send projection fibres to the ventral striatum and the autonomic/limbic-associated areas of the prefrontal cortex. In the present study, we sought to determine whether or not single midline thalamic neurons project both to the ventral striatum and to the cerebral cortical areas. Experiments were performed on chloral hydrate-anaesthetized male Sprague Dawley rats; two fluorescent retrograde tracers were centred on the medial or lateral part of the nucleus accumbens--the major part of the ventral striatum--and the medial or lateral prefrontal viscerolimbic cortex. Our retrograde double-labelling study revealed that a subset of midline thalamic neurons send projection fibres to both the nucleus accumbens and the cerebral cortex. Such neurons projecting to both targets were principally identified in the paraventricular thalamic nucleus. The majority of the dually-labelled neurons in the paraventricular thalamic nucleus projected to the lateral part of the nucleus accumbens and the medial wall of the prefrontal cortex. Dually-labelled neurons were additionally found in other midline nuclei, including the paratenial, intermediodorsal, rhomboid, and reuniens nuclei, as well as in the medial part of the parafascicular thalamic nucleus. Dually-projecting neurons identified in the present study may represent a potential link between the limbic striatum and the viscerolimbic-associated cortex, thus suggesting that non-discriminative information relayed to the prefrontal cortex might exert an influence through the same neurons on the nucleus accumbens implicated in affective behaviour.


Our experience with transanal endorectal pull-through in Hirschsprung's disease.
The aim of the study was to evaluate the functional results, complications, the problems caused, and the outcomes of the transanal endorectal pull-through (TEPT) operation in Hirschsprung's disease. The 22 patients who were operated for Hirschsprung's disease with TEPT between November 2003 and September 2006 were reviewed retrospectively. The patients were evaluated for age, gender, operational findings, duration of hospitalization and functional outcomes after the operation. A total of 22 patients, sixteen males (72.7 %) and 6 females (27.3 %) aged 23 days to 11 years (mean 19.3 +/- 6.9 months), were operated using TEPT over a 34-month period. The mean length of the resected aganglionic segment was 23 +/- 2.4 cm; the shortest segment was 7 cm and the longest 40 cm. The postoperative hospital stay was 3 - 10 days, oral feeding was started at 1 - 4 days, the first bowel movement was at 1 - 7 days and the number of daily movements for patients in whom the colostomy was closed was 2 - 5. The mean postoperative follow-up period was 18 +/- 2.4 months (1 - 33 months). Two patients (9 %) were hospitalized once for enterocolitis. One patient had a constipation problem that resolved with medical treatment. One patient needed colostomy for anastomosis leakage on the 5th postoperative day, followed by a redo pull-through using a posterior sagittal approach. None of the patients had a continence problem. No urethral damage was observed and there were no abscesses at the muscular cuff. We observed that mucosal dissection was more difficult in the rectal biopsy area. Although only recently accepted, TEPT has quickly found a place in clinical practice as it is based on an operational technique whose results are well identified and accepted and with which there is extensive experience. It seems that TEPT has the advantages of having no additional problems compared to the classical techniques with respect to complications and functional outcomes while providing better patient comfort and cosmetic outcomes. We conclude that TEPT may be preferred in appropriate cases and will evolve to become a more practical and effective technique.


The Relationship Between APOE Gene Polymorphism and In-stent Restenosis After Stenting at the Beginning of the Vertebral Artery.
To retrospectively investigate the relationship between apolipoprotein E (APOE) gene polymorphism and in-stent restenosis (ISR) after stenting at the beginning of the vertebral artery. The study included 155 patients who successfully underwent stenting at the beginning of the vertebral artery and had postoperative digital subtraction angiography or computed tomography angiography. Based on the follow-up results, they were divided into the restenosis (ISR) group and non-restenosis (non-ISR) group. The clinical information and APOE genotypes of both groups were analyzed. A binary logistic regression model was used to analyze independent risk factors for ISR. After 1 year of follow-up, 49 (31.6%) patients had ISR and 106 (68.4%) did not. Binary logistic regression analysis showed that serum low-density lipoprotein cholesterol (LDL-C), serum lipoprotein-related phospholipase A2 (Lp-PLA2), and E3/E4 genotype were independent risk factors for ISR after stenting at the beginning of the vertebral artery. In addition, the LDL-C level of patients with the E3/E4 genotype was higher compared with the E3/E3 genotype group (P < 0.05). APOE gene polymorphism is associated with ISR, and the E3/E4 genotype is an independent risk factor for ISR after stenting at the beginning of the vertebral artery. Further genetic studies can identify risk genotypes to facilitate the early prediction and identification of high-risk patients with ISR.


Brief report: Neighborhood disadvantage and hair cortisol among older urban African Americans.
Previous studies have shown that living in poor neighborhoods is associated with increased morbidity and mortality. However, researchers are now investigating the biological pathways responsible for the deleterious effects of neighborhood disadvantage on health. This study investigated whether neighborhood disadvantage (i.e., a measure of relative neighborhood quality derived by combining social and built environmental conditions) was associated with hair cortisol-a retrospective indicator of long-term hypothalamic pituitary adrenal (HPA) axis activation-and whether this link would be mediated by self-reported neighborhood satisfaction. Forty-nine older African Americans were recruited from thirty-nine Detroit census tracts across five strata of census tract adversity. Participants were interviewed face-to-face to collect psychosocial measures. Each provided a hair sample for analysis of cortisol. Multiple regression analyses revealed that higher neighborhood disadvantage was associated with higher levels of hair cortisol levels and that neighborhood satisfaction partially explained this association. These results are the first to our knowledge to demonstrate a direct link between neighborhood disadvantage and hair cortisol in a sample of older adults and to show that self-reported neighborhood satisfaction may be a psychological intermediary of this association.


The role of teachers and schools in explaining STEM outcome gaps.
We develop a novel methodology to explore the extent to which teacher and school assignments between fourth and eighth grade contribute to inequalities between advantaged and disadvantaged students-as defined by underrepresented minority (URM) status and eligibility for free or reduced-price lunch (FRL)-in their eighth-grade math and science test scores and high school math and science course-taking. We find that differences between advantaged and disadvantaged students in teacher and school assignments predict about 25% of the eighth-grade math test gaps, 10% of the eighth-grade science test gaps, and 35% of the gaps in advanced math course-taking. For science course-taking, school and teacher assignments predict about 20% of the URM gap and 10% of the FRL gap. While this methodology does not permit causal conclusions, the results are robust to analyses of a subsample in which students do not appear to be assigned to teachers based on prior test achievement.


Unloading work of breathing during high-frequency oscillatory ventilation: a bench study.
With the 3100B high-frequency oscillatory ventilator (SensorMedics, Yorba Linda, CA, USA), patients' spontaneous breathing efforts result in a high level of imposed work of breathing (WOB). Therefore, spontaneous breathing often has to be suppressed during high-frequency oscillatory ventilation (HFOV). A demand-flow system was designed to reduce imposed WOB. An external gas flow controller (demand-flow system) accommodates the ventilator fresh gas flow during spontaneous breathing simulation. A control algorithm detects breathing effort and regulates the demand-flow valve. The effectiveness of this system has been evaluated in a bench test. The Campbell diagram and pressure time product (PTP) are used to quantify the imposed workload. Using the demand-flow system, imposed WOB is considerably reduced. The demand-flow system reduces inspiratory imposed WOB by 30% to 56% and inspiratory imposed PTP by 38% to 59% compared to continuous fresh gas flow. Expiratory imposed WOB was decreased as well by 12% to 49%. In simulations of shallow to normal breathing for an adult, imposed WOB is 0.5 J l-1 at maximum. Fluctuations in mean airway pressure on account of spontaneous breathing are markedly reduced. The use of the demand-flow system during HFOV results in a reduction of both imposed WOB and fluctuation in mean airway pressure. The level of imposed WOB was reduced to the physiological range of WOB. Potentially, this makes maintenance of spontaneous breathing during HFOV possible and easier in a clinical setting. Early initiation of HFOV seems more possible with this system and the possibility of weaning of patients directly on a high-frequency oscillatory ventilator is not excluded either.


Upregulation of high-affinity GABA(A) receptors in cultured rat dorsal root ganglion neurons.
Despite evidence that high-affinity GABA(A) receptor subunit mRNA and protein are present in dorsal root ganglia (DRG), low-affinity currents dominate those detected in acutely dissociated DRG neurons in vitro. This observation raises the possibility that high-affinity receptors are normally trafficked out of the DRG toward central and peripheral terminals. We therefore hypothesized that with time in culture, there would be an increase in high-affinity GABA(A) currents in DRG neurons. To test this hypothesis, we studied dissociated DRG neurons 2 h (acute) and 24 h (cultured) after plating with whole-cell patch-clamp techniques, Western blot, and semiquantitative reverse transcriptase polymerase chain reaction (sqRT-PCR) analysis. GABA(A) current density increases dramatically with time in culture in association with the emergence of two persistent currents with EC50's of 0.25±0.01 μM and 3.2±0.02 μM for GABA activation. In a subpopulation of neurons, there was also an increase in the potency of GABA activation of the transient current from an EC50 of 78.16±10.1 μM to 9.56±1.3 μM with time in culture. A fraction of the high-affinity current was potentiated by δ-subunit agonist 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridine-3-ol (THIP). δ-subunit immunoreactivity was largely restricted to the cytosolic fraction in acute, but the membrane fraction in cultured, DRG neurons, with no detectable change in δ-subunit mRNA. However, the emergence of a high-affinity current blocked by THIP and insensitive to bicuculline was detected in a subpopulation of cultured neurons as well in association with an increase in ρ2- and ρ3-subunit mRNA in cultured DRG neurons. Our results suggest that high-affinity δ-subunit-containing GABA(A) receptors are normally trafficked out of the DRG where they are targeted to peripheral and central processes. They also highlight that the interpretation of data obtained from cultured DRG neurons should be made with caution.


A highly branched α-d-glucan facilitates antitumor immunity by reducing cancer cell CXCL5 expression.
Tumor immunotherapy has emerged as a major pillar of anticancer therapeutic strategies. Natural polysaccharides, known for their strong immunomodulatory activities with relatively low cost and toxicity, are becoming promising prospects for cancer immunotherapy. In this study, we investigated the antitumor mechanism of JNY2PW, a highly branched α-d-glucan previously purified from the traditional marine Chinese medicine Arca inflata. JNY2PW was shown to enhance the sensitivity of tumor cells to co-culture macrophage supernatants by decreasing cancer cell CXCL5 expression. Furthermore, JNY2PW exerted antitumor effects without obvious toxic side effects in tumor-bearing mice by triggering the Akt/mTOR and ERK/GSK3β/β-catenin pathways and attenuating expression of CXCL5 in cancer cells. Remarkably, JNY2PW reduced tumor proliferation and dampened CXCL5 expression in tumor cells overexpressing CXCL5 both in vitro and in vivo. Additionally, JNY2PW blocked epithelial-mesenchymal transition (EMT) in both CXCL5-overexpressing and wild type tumor cells. Our data therefore uncovered a previously unrecognized antitumor mechanism for JNY2PW, suggesting that JNY2PW is a promising adjuvant as an immunomodulator for cancer immunotherapy.


Are animal models useful for understanding the pathophysiology of lysosomal storage disease?
Spontaneously occurring genetic lysosomal storage diseases are as rare in other mammalian species as in man. However, the advent of gene targeting technology has revolutionized the state of animal models of genetic diseases. Nearly all lysosomal storage diseases known in man have been duplicated in the mouse. The technology now allows, not only complete inactivation of endogenous genes, but also the introduction of essentially any type of mutation. These animal models can overcome many of the limitations inherent in studies of human patients--rarity of the disease, extremely complex genetic background and logistical and ethical constraints in the design and execution of experiments with human subjects. For example, genetic manipulations of germ cells or cross-breeding experiments between two mutants are readily feasible with animal models. Two major areas of the utility of animal models are the clarification of the pathophysiology/pathogenetic mechanism of disease and the exploration of therapeutic approaches. Examples of experiments using animal models of lysosomal storage disease are presented, primarily from studies undertaken in our own laboratory. Animal models have proved invaluable in extending our knowledge of the lysosomal storage diseases and exploring potential therapies.


Durability evaluation of biopolymer coating on titanium alloy substrate.
For this study, a commercially available phosphorylcholine (PC) polymer was applied to Ti6Al4V ELI. A multivariate approach to design a statistically significant array of experiments was employed to evaluate and estimate optimization of PC-immobilization process factors. The seven process factors analyzed were (1) power level for RFGD plasma treatment, (2) duration of plasma treatment, (3) concentration of PC solution used to coat samples, (4) rate at which samples were dipped in/out of the solution, (5) temperature for curing, (6) relative humidity level during curing, and (7) duration of curing. Imaging and analysis of the coating were done via fluorescence microscopy (FM), confirming the uniform coverage of PC polymer on titanium substrate. The process factors were evaluated by three measured responses: initial thickness, coating durability and degree of cross-linked coating, which were assessed by FM, a spray test and extraction in IPA, respectively. Variations in PC solution concentration showed no impact on fouling resistance of the resultant coating. It was hypothesized that the PC-application process factors could be optimized to yield favorable outcomes in durability and degree of cross-linked coating responses. The resulting statistical model indicates that PC solution concentration, dip rate, and cure temperature are the three greatest singular effects on both durability and degree of cross-linking. In addition, plasma treatment of the substrate with O2 was effective in enhancing the degree of cross-linking of the polymer surface.


Sleeve lobectomy in patients with non-small-cell lung cancer: a report from the European Society of Thoracic Surgery database 2021.
For centrally located lung tumours, sleeve lobectomy is preferred over pneumectomy. We report on the surgical practices and perioperative outcomes of sleeve resections based on data from the European Society of Thoracic Surgeons database. We retrieved data of patients undergoing sleeve lobectomy or bilobectomy from 2007 to 2021. We evaluated baseline characteristics, surgical approach, neoadjuvant treatments, morbidity and postoperative outcomes of open and video-assisted thoracoscopic surgery (VATS) procedures. In total, 1652 patients (median age: 63 years; females/males: 446/1206) underwent sleeve lobectomy (n = 1536) or bilobectomy (n = 116) by open thoracotomy (n = 1491; 90.2%) or VATS (n = 161; 9.8%) with a thoracotomy conversion rate of 21.1% (n = 34); 398 (24.1%) patients received neoadjuvant treatment. Overall morbidity and 30-day mortality were 40.6% and 2.2%, respectively. Bronchial anastomotic complications occurred in 29 patients (1.8%) with conservative treatment in 6 cases (20.7%) and operative management in 23 (79.3%). On multivariable analysis, factors related to the elevated risk of cardiopulmonary complications were body mass index < 20 [odds ratio (OR): 2.26; P < 0.001] and bilobectomy (OR : 2.28, P < 0.001). Age <60 years (OR: 0.71, P = 0.013), female sex (OR: 0.54, P < 0.001) and VATS (0.64, P < 0.001) were associated with decreased risk. Neoadjuvant treatment was not associated with increased risks of cardiopulmonary complications (OR: 1.05; P = 0.664). Compared to open thoracotomy, VATS was associated with significantly decreased overall morbidity (30.4% vs 41.7%, P = 0.006) and length of stay (median: 5 days vs 8 days; P < 0.001). Sleeve lobectomies can be safely performed after neoadjuvant treatment. The VATS approach fosters shorter length of stay and decreased morbidity.


Ficus crocata leaf extracts decrease the proliferation and invasiveness of breast cancer cells.
Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype due to its greater invasive capacity and non-response to hormone therapy. Several species of the Ficus genus have been used as an alternative to traditional medicine against malignant diseases. Previously, leaf extracts from Ficus crocata (Miq.) Mart. ex Miq. (F. crocata) showed antiproliferative activity in vitro against breast and cervical tumor cells without having a cytotoxic effect on non-tumor cell lines. The purpose of the study was to evaluate the effect of hexane (Hex-EFc), dichloromethane (Dic-EFc), and acetone (Ace-EFc) extracts from F. crocata on the proliferative and invasive capacity of breast cancer cells MCF-7 and MDA-MB-231. The phytochemical profile was carried out by gas chromatography-mass spectrometry (GC-MS). Cell proliferation, migration, and invasion were determined by MTT, wound closure, and transwell assays, respectively. MMPs activity was analyzed using gelatin zymography, and fluorescence microscopy was used to visualize F-actin distribution. Hex-EFc, Dic-EFc, and Ace-EFc showed cytotoxic activity on MDA-MB-231 tumor cells and, to a lesser extent, on MCF-7 cells, without presenting cytotoxicity at the same concentrations in MCF-10A non-tumor cells. Dic-EFc and Ace-EFc (5-10 μg/mL) reduced the migration capacity of MCF-7 and MDA-MB-231 cells. Interestingly, exposure to Dic-EFc and Ace-EFc (5-10 μg/mL) inhibited the invasive ability of MDA-MB-231 cells, reducing the secretion and activity of MMP-2 and MMP-9, as well as the F-actin distribution. Dic-EFc and Ace-EFc at low concentrations decreased breast cancer cell proliferation and invasiveness, mainly of MDA-MB-231 cells. The above supports the potential use of compounds from leaf extracts of F. crocata in neoadjuvant therapy to reduce the progression of breast cancer tumors, mainly triple-negative tumors.


Breakthrough instruments and products: Lattice Lightsheet 7-Non-invasive imaging of the subcellular dynamics of life.
Light sheet fluorescence microscopy has become an established method for fast and sensitive imaging of living specimens with minimum phototoxicity and photobleaching. By adding lattice structures to the light sheet, the ZEISS Lattice Lightsheet 7 makes this technique available for live cell imaging at subcellular resolution while also allowing microscopists to use their standard sample carriers.


Endogenous lateral electric fields around bovine corneal lesions are necessary for and can enhance normal rates of wound healing.
The strength of the electric fields in the vicinity of 1.5 mm circular lesions in the bovine cornea has been found to influence the rate of re-epithelization. A decrease in the field strength by submersion of the lesions or by treating the lesions with the Na+-channel blocker, benzamil, significantly retarded healing. An increase in the field strength of lesions treated with Na+-depleted Hanks' solution, by the addition of direct current, increased epithelization. Epithelization was fastest in wounds with field strengths raised to - 80 mV/mm, more than twice the normal field strength present in wounds maintained in Hanks' solution alone. Epithelization decreased, however, when the field strengths were increased to -120 mV/mm. A similar pattern was also observed when the field's polarity was reversed. By manipulating and monitoring the field strengths, we have been able to show for the first time that increased wound field strengths enhance corneal wound epithelization, and that field strengths with reversed polarity also enhance this epithelization.


Competitive interaction between two functional S-haplotypes confer self-compatibility on tetraploid Chinese cherry (Prunus pseudocerasus Lindl. CV. Nanjing Chuisi).
Self-incompatibility (SI) has been studied extensively at the molecular level in Solanaceae, Rosaceae and Scrophulariaceae, all of which exhibit gametophytic self-incompatibility (GSI). In the present study, four PpsS-haplotypes (Prunus pseudocerasus S-haplotypes) comprising at least two genes, i.e., PpsS-RNase (P. pseudocerasus S-RNase) and PpsSFB (P. pseudocerasus S-haplotype-specific F-box) have been successfully isolated in tetraploid P. pseudocerasus Lindl. CV. Nanjing Chuisi ("NC") which exhibited self-compatibility (SC), and its S-genotype was determined as S-1/S-3'/S-5/S-7. These PpsS-RNases, which were expressed exclusively in style, shared the typical structural features with S-RNases from other Prunus species exhibiting GSI. All PpsSFBs showed similar structure characteristics of SFBs from other Prunus species, and matched with the necessary conditions for pollen S-determinant. No mutations leading to dysfunction of S-haplotype were found in their full-length c-DNA sequences, except for PpsS-3'-haplotype which was not amplified by PCR. These four S-haplotypes complied with tetrasomic inheritance. Diploid pollen grains with S-genotypes S-7/S-1, S-7/S-5 and S-1/S-5 can grow the full length of the style after self-pollination, while pollen grains with S-3'/S-7, S-3'/S-1 and S-3'/S-5 cannot. These results suggest that PpsS-haplotypes-1, -5 and -7 are functional, and that competitive interaction between two of them confer self-compatibility on cultivar "NC". Furthermore, in terms of recognition specificity, diploid pollen grains carrying PpsS-3'-haplotype are equal to monoploid pollen grains carrying the other functional S-haplotype.


Optical control of filamentation-induced damage to DNA by intense, ultrashort, near-infrared laser pulses.
We report on damage to DNA in an aqueous medium induced by ultrashort pulses of intense laser light of 800 nm wavelength. Focusing of such pulses, using lenses of various focal lengths, induces plasma formation within the aqueous medium. Such plasma can have a spatial extent that is far in excess of the Rayleigh range. In the case of water, the resulting ionization and dissociation gives rise to in situ generation of low-energy electrons and OH-radicals. Interactions of these with plasmid DNA produce nicks in the DNA backbone: single strand breaks (SSBs) are induced as are, at higher laser intensities, double strand breaks (DSBs). Under physiological conditions, the latter are not readily amenable to repair. Systematic quantification of SSBs and DSBs at different values of incident laser energy and under different external focusing conditions reveals that damage occurs in two distinct regimes. Numerical aperture is the experimental handle that delineates the two regimes, permitting simple optical control over the extent of DNA damage.


A novel STAT3 mutation associated with hyper immunoglobulin E syndrome with a paucity of connective tissue signs.
A heterozygous mutation of STAT3 causes autosomal dominant hyper immunoglobulin E (IgE) syndrome; however, there are still many unclear points regarding the clinical spectrum of this syndrome. In addition to a clinical description of patients in terms of pedigree, a genetic analysis, quantitation of peripheral blood Th17 and ex vivo IL-17 production were carried out. The proband, a 2-year-old boy (Patient 1) with early onset atopic dermatitis-like eczema and recurrent bacterial infections, was suspected of autosomal dominant hyper immunoglobulin E syndrome on the basis of his symptoms and family history. His mother (Patient 2) also had skin eczema and recurrent bacterial infections, and his sister (Patient 3) had skin eczema. A novel STAT3 mutation (p.S476F) was detected in all three patients, but not in the father, who had no such symptoms. A significant decrease in peripheral blood Th17 subsets and IL-17 production was found in all the patients. Curiously, all three patients carrying the p.S476F mutation in STAT3 lacked connective tissue signs such as distinctive facial features, retention of primary teeth, and joint hyperextensibility. Autosomal dominant hyper IgE syndrome should, perhaps, be considered even if patients lack connective tissue signs, as long as hypersensitivity to infection and skin manifestations with hyper IgE are present.


Ultrathin dendrimer-graphene oxide composite film for stable cycling lithium-sulfur batteries.
Lithium-sulfur batteries (Li-S batteries) have attracted intense interest because of their high specific capacity and low cost, although they are still hindered by severe capacity loss upon cycling caused by the soluble lithium polysulfide intermediates. Although many structure innovations at the material and device levels have been explored for the ultimate goal of realizing long cycle life of Li-S batteries, it remains a major challenge to achieve stable cycling while avoiding energy and power density compromises caused by the introduction of significant dead weight/volume and increased electrochemical resistance. Here we introduce an ultrathin composite film consisting of naphthalimide-functionalized poly(amidoamine) dendrimers and graphene oxide nanosheets as a cycling stabilizer. Combining the dendrimer structure that can confine polysulfide intermediates chemically and physically together with the graphene oxide that renders the film robust and thin (<1% of the thickness of the active sulfur layer), the composite film is designed to enable stable cycling of sulfur cathodes without compromising the energy and power densities. Our sulfur electrodes coated with the composite film exhibit very good cycling stability, together with high sulfur content, large areal capacity, and improved power rate.


Structural changes in nucleoli during inhibition of protein- and RNA-biosynthesis.
A variety of cells (unstimulated human lymphocytes, phytohemagglutinin-stimulated human lymphocytes, diploid human fibroblasts, human melanoma cells, and Hela cells) were subjected in vitro to inhibition of protein biosynthesis by puromycin. Hela cells were also treated with actinomycin D to inhibit RNA-synthesis. Under puromycin treatment, the fibrillar centers of the nucleoli were smaller in all actively dividing cell types, whereas in small inactive lymphocytes from peripheral blood the inhibition of protein synthesis had no noticeable effect. Nucleoli with nucleolonema changed into compact nucleoli under puromycin treatment. When RNA-synthesis was inhibited, the fibrillar centers remained at an approximately constant volume. These findings indicate that proteins localized in the fibrillar centers are involved in, and are used up during, rDNA-transcription and/or further steps of ribosome biogenesis. The changes in nucleolar architecture after the inhibition of protein synthesis suggest that transcriptional processes become concentrated near sites where proteins have been stored, i.e. the fibrillar centers.


It Is a Brave New World: Alternative Payment Models and Value Creation in Total Joint Arthroplasty: Creating Value for TJR, Quality and Cost-Effectiveness Programs.
The increasing cost of our country's healthcare is not sustainable. To address this crisis, the federal government is transiting healthcare reimbursement from the traditional volume-based system to a value-based system. As such, increasing healthcare value has become an essential point of discussion for all healthcare stakeholders. The purpose of this study is to discuss the importance of healthcare value as a means to achieve this goal of value-based medicine and 3 methods to create value in total joint arthroplasty. These methods are to: (1) improve outcomes greater than the increased costs to achieve this improvement, (2) decrease costs without affecting outcomes, and (3) decrease costs while simultaneously improving outcomes. Following these guidelines will help practitioners thrive in a bundled care environment.


Right Ventricular Strain to Assess Early Right Heart Failure in the Left Ventricular Assist Device Candidate.
Right heart failure (RHF) following left ventricular assist device implantation (LVAD) remains the primary cause of postoperative mortality and morbidity, and prediction of RHF is the main interest of the transplantation community. In this review, we outline the role and impact of right ventricular strain in the evaluation of the right ventricle function before LVAD implantation. Accumulating data suggest that measurement of right ventricular longitudinal strain (RVLS) has a critical role in predicting RHF preoperatively and may improve morbidity and mortality following LVAD implantation. However, the significant intraobserver, interobserver variability, the lack of multicenter, prospective studies, and the need for a learning curve remain the most critical limitations in the clinical practice at present. This review highlighted the importance of right ventricular strain in the diagnosis of RHF preoperatively and revealed that RVLS might have a crucial clinical measurement for the selection and management of LVAD patients in the future with the more extensive multicenter studies.


Effects of obesity and acute resistance exercise on skeletal muscle angiogenic communication pathways.
What is the central question of this study? Do obesity and acute resistance exercise alter the regulation of muscle intercellular communication pathways consistent with inadequate compensatory angiogenesis in response to muscle loading present in individuals with obesity? What is the main finding and its importance? Obesity is associated with differences in both pro- and anti-angiogenic signalling consistent with lower muscle capillarization. Acute resistance exercise increases the release of skeletal muscle small extracellular vesicles independent of body mass. These results identify new cellular factors associated with impaired angiogenesis in obesity and the positive effects of acute resistance exercise in lean and obese skeletal muscle. Obesity (OB) impairs cell-to-cell communication signalling. Small extracellular vesicles (EVs), which include exosomes, are released by skeletal muscle and participate in cell-to-cell communication, including the regulation of angiogenesis. Resistance exercise (REx) increases muscle fibre size and capillarization. Although obesity increases muscle fibre size, there is an inadequate increase in capillarization such that capillary density is reduced. It was hypothesized that REx-induced angiogenic signalling and EV biogenesis would be lower with obesity. Sedentary lean (LN) and OB subjects (n = 8 per group) performed three sets of single-leg knee-extension REx at 80% of maximum. Muscle biopsies were obtained at rest, 15 min and 3 h postexercise and analysed for angiogenic and EV biogenesis mRNA and protein. In OB subjects, muscle fibre size was ∼20% greater and capillary density with type II fibres ∼25% lower compared with LN subjects (P < 0.001). In response to REx, the increase in VEGF mRNA (pro-angiogenic) was similar (3-fold) between groups, while thrombospondin-1 (TSP-1) mRNA (anti-angiogenic) increased ∼2.5-fold in OB subjects only (P = 0.010). miR-130a (pro-angiogenic) was ∼1.4-fold (P = 0.011) and miR-503 (anti-angiogenic) ∼1.8-fold (P = 0.017) greater in OB compared with LN subjects at all time points. In both groups, acute REx decreased the EV surface protein Alix by ∼50%, consistent with the release of exosomes (P = 0.016). Acute REx appears to induce the release of skeletal muscle small EVs independent of body mass. However, with obesity there is predominantly impaired angiogenic signalling, consistent with inadequate angiogenesis in response to basal muscle hypertrophy.


Controlled trial of furosemide therapy in infants with chronic lung disease.
To study the effects of furosemide therapy in infants with chronic lung disease (CLD), a double-blind controlled trial was designed. Seventeen infants with evidence of CLD (oxygen requirements greater than 30% at greater than 3 weeks of age and chest radiographic findings consistent with CLD) were studied. Pulmonary function was measured immediately before, and after 48 hours and 7 days of treatment with furosemide (1 mg/kg/12 hr intravenously or 2 mg/kg/12 hr orally) or placebo. Clinical status improved in six of seven infants who received furosemide and in two of 10 infants who received placebo (P less than 0.002). In the furosemide group, ventilator and oxygen requirements decreased (P less than 0.003); minute ventilation, alveolar ventilation, and dynamic compliance increased; and venous admixture decreased (P less than 0.05). There were no significant changes in the placebo group. Our findings suggest that furosemide significantly improves lung function during therapy in infants with CLD and allows earlier weaning from ventilatory support and supplemental oxygen.


Unexpected Septal and Truncal Valve Endocarditis After Transcatheter Pulmonary Valve Replacement.
Despite early clinical success of transcatheter pulmonic valve replacement, there is concern for an increased risk of endocarditis requiring complex surgery to repair. We present a case of endocarditis of a Melody (Medtronic, Minneapolis, MN) valve in a 33-year-old male patient with prior neonatal repair of persistent truncus arteriosus and 2 subsequent right ventricular outflow tract to pulmonary artery conduit replacements. The infection had extended from the Melody valve through the prior ventricular septal defect patch to the truncal valve and highlights the risk of endocarditis, particularly with the Melody transcatheter valve.


Human proinsulin C-peptide prevents proliferation of rat aortic smooth muscle cells cultured in high-glucose conditions.
Proinsulin C-peptide is involved in several biological activities. However, the role of C-peptide in vascular smooth muscle cells is unclear. We therefore investigated its effects, in vascular smooth muscle cells in high-glucose conditions. Rat aortic smooth muscle cells were cultured with 5.5 or 20 mmol/l glucose with or without C-peptide (1 to 100 nmol/l) for 3 weeks. Proliferation activities, the protein expression of platelet-derived growth factor (PDGF)-beta receptor, the phosphorylation of p42/p44 mitogen-activated protein (MAP) kinases, and glucose uptake were measured. The proliferation activities increased approximately three-fold under high-glucose conditions (p<0.05). C-peptide suppressed hyperproliferation activities that were induced by high glucose. This happened in a dose-dependent manner from 1 to 100 nmol/l of C-peptide. C-peptide (10 and 100 nmol/l) inhibited the increased protein expression of PDGF-beta receptor and the phosphorylation of p42/p44 MAP kinases that had been induced by high glucose (p<0.05). Furthermore, 100 nmol/l of C-peptide augmented the impaired glucose uptake in the high-glucose conditions. These observations suggest that C-peptide could prevent diabetic macroangiopathy by inhibiting smooth muscle cell growth and ameliorating glucose utilisation in smooth muscle cells. C-peptide may thus be a novel agent for treating diabetic macroangiopathy in patients with type 1 and type 2 diabetes.


[Effect of sinigrin on sucrose uptake by some polyphagous and oligophagous aphids (Aphididae)].
The effect of 0.1% sinigrin on sucrose uptake (20% aqueous solution) by some polyphagous and oligophagous aphid species was tested using the tracer method. In the presence of sinigrin the sucrose ingestion of Brevicoryne brassicae, Myzus persicae and Megoura viciae was promoted. With Rhopalosiphum padi there was a strong, and with Aphis fabae and Acyrthosiphon pisum only a slight inhibition of sucrose uptake. Aulacorthum circumflexum reacted positively in some experiments, but negatively in others. Thus the acceptance or refusal of cruciferous plants as a host by the majority of the species examined corresponded to their reaction to sinigrin. Contrasting results were obtained with M. viciae. Although cruciferous plants are non hosts for this species, it ingested more sucrose when sinigrin was present. Therefore other physical or chemical properties must be considered for the refusal of cruciferous plants as a host.


Lipomatous hypertrophy of the interatrial septum: A potential pitfall resolved on comparing previous PET/CT.
In this case report, we describe a rare case of inter-atrial septal lipomatosis occasionally found in a PET-CT scan performed in an 81-year-old patient with a history of periprosthetic endocarditis post aortic prosthesis implantation. The patient reappeared in 2022 in the emergency department complaining of symptomatology of worsening asthenia, fever, and elevated inflammatory indices. He was hospitalized in Cardiology department on suspect of recurrence of endocarditis and underwent PET-CT examination that showed an increased metabolic finding at the interatrial septum. On possible suspicion of recurrence of infective endocarditis or cardiac tumor pathology, further diagnostic investigation by cardio-RM examination was requested. However, after radiologic consultation, the previous performed FDG-PET/TC examinations were re-evaluated and a misdiagnosed uptake at this level was revealed, which was assumed to be due to the endocarditis condition because of widespread uptake throughout the perivalvular area.


Identification of the novel HLA-DQB1*03:483 allele by sequencing-based typing.
A single nucleotide change in exon 1 of HLA-DQB1*03:01:01:03 results in the novel HLA-DQB1*03:483 allele.


A rationale for repression and/or loss of motility by pathogenic Yersinia in the mammalian host.
Pathogenic yersiniae either repress flagella expression under host conditions (Yersinia enterocolitica and Yersinia pseudotuberculosis) or have permanently lost this capability by mutation (Yersinia pestis). The block in flagella synthesis for the enteropathogenic Yersinia centers on fliA (sigmaF) repression. This repression ensures the downstream repression of flagellin structural genes which can be cross-recognized and secreted by virulence type III secretion systems. Y. pestis carries several flagellar mutations including a frame shift mutation in flhD, part of the flagellar master control operon. Repression of flagellins in the host environment may be critical because they are potent inducers of innate immunity. Artificial expression of flagellin in Y. enterocolitica completely attenuates virulence, supporting the hypothesis that motility is a liability in the mammalian host.


Trends in spontaneous adverse drug reaction reports to the French pharmacovigilance system (1986-2001).
The French pharmacovigilance system is based on a network of 31 regional centres located in teaching hospitals and coordinated by the French Medicines Agency ("Agence Française de Sécurité Sanitaire des Produits de Santé" [Afssaps]). Since 1984, they have shared a common database of adverse drug reactions (ADRs) that are spontaneously reported by healthcare professionals. The objective of this study is to describe the characteristics of the reports and the reporting trends in the French pharmacovigilance spontaneous reporting database from 1986 to 2001. All the reports from January 1986 to December 2001 were included. Drugs and ADRs were translated to anatomical therapeutic chemical (ATC) codes and MedDRA classifications, respectively. The total number of reports was 197 580 over the 16-year period, with linearly increase over time. The median (interquartile range [IQR]) age of the patients was 53 (34-70) and the male/female ratio was 0.82. The median (IQR) time between the date of occurrence of the ADR and the date of report was 73 days (34-166). The reporter was a specialist in 74% of the reports and a general practitioner in 17%. The annual rate of reporting according to medical demography strongly increased for the specialists, especially since 1994. At least one ADR was considered as serious in 44.8% of the reports. The ADRs were most frequently related to nervous system drugs (23%), followed by cardiovascular drugs (19%) and systemic anti-infectives (17%). The latter class had the fastest progression mostly due to antiretroviral therapy since 1996. According to the Medical Dictionary for Regulatory Activities (MedDRA) coding, the system organ most often reported was skin and subcutaneous tissue disorders (29%), followed by nervous system disorders (19%), gastrointestinal disorders (12%), blood and lymphatic system disorders (12%), vascular disorders (12%) and general disorders and administration site conditions (12%). All spontaneous reporting systems are affected by under-reporting. One of their goals is to generate early signals, which might be more affected by reporting bias than by under-reporting. Some improvements should be made in the design of the French database, but data collected since 1986 constitute an essential tool for the routine work of the 31 pharmacovigilance centres. This first description of the data of the French pharmacovigilance database involving all drugs and ADRs shows an increasing tendency to reporting over time, especially in specialists and for systemic anti-infective drugs. The database that uses hierarchical international classifications for drugs and adverse reactions may be used for further studies and could be the basis for an automatic signal generation system.