CXCL12-mediated murine neural progenitor cell movement requires PI3Kβ activation.
The migratory route of neural progenitor/precursor cells (NPC) has a central role in central nervous system development. Although the role of the chemokine CXCL12 in NPC migration has been described, the intracellular signaling cascade involved remains largely unclear. Here we studied the molecular mechanisms that promote murine NPC migration in response to CXCL12, in vitro and ex vivo. Migration was highly dependent on signaling by the CXCL12 receptor, CXCR4. Although the JAK/STAT pathway was activated following CXCL12 stimulation of NPC, JAK activity was not necessary for NPC migration in vitro. Whereas CXCL12 activated the PI3K catalytic subunits p110α and p110β in NPC, only p110β participated in CXCL12-mediated NPC migration. Ex vivo experiments using organotypic slice cultures showed that p110β blockade impaired NPC exit from the medial ganglionic eminence. In vivo experiments using in utero electroporation nonetheless showed that p110β is dispensable for radial migration of pyramidal neurons. We conclude that PI3K p110β is activated in NPC in response to CXCL12, and its activity is necessary for immature interneuron migration to the cerebral cortex.


Influenza virus resistance to neuraminidase inhibitors: implications for treatment.
Oseltamivir and Zanamivir are the two main Neuraminidase inhibitors used for the treatment of Influenza. Oseltamivir resistance has been identified in non-pandemic influenza viruses, as well as H1N1 pandemic Influenza A viruses. Resistance is associated with increased morbidity, and poorer outcomes in severely immunocompromised hosts. Newer neuraminidase inhibitors, increased vaccination and combination therapy may be alternatives for the treatment of Influenza in this setting.


Regulation of vincamine biosynthesis and associated growth promoting effects through abiotic elicitation, cyclooxygenase inhibition, and precursor feeding of bioreactor grown Vinca minor hairy roots.
Hydroxylase/acetyltransferase elicitors and cyclooxygenase inhibitor along with various precursors from primary shikimate and secoiridoid pools have been fortified to vincamine less hairy root clone of Vinca minor to determine the regulatory factors associated with vincamine biosynthesis. Growth kinetic studies revealed that acetyltransferase elicitor acetic anhydride and terpenoid precursor loganin significantly reduce the growth either supplemented alone or in combination (GI = 140.6 ± 18.5 to 246.7 ± 24.3), while shikimate and tryptophan trigger biomass accumulation (GI = 440.2 ± 31.5 to 540.5 ± 40.3). Loganin also downregulates total alkaloid biosynthesis. Maximum flux towards vincamine production (0.017 ± 0.001 % dry wt.) was obtained when 20-day-old hairy roots were fortified with secologanin (10 mg/l) along with tryptophan (100 mg/l), naproxen (8.4 mg/l), hydrogen peroxide (20 μg/l), and acetic anhydride (32.4 mg/l). This was supported by RT PCR (qPCR) analysis where 2- and 3-fold increase in tryptophan decarboxylase (TDC; RQ = 2.0 ± 0.09) and strictosidine synthase (STR; RQ = 3.3 ± 0.36) activity, respectively, was recorded. The analysis of variance (ANOVA) for growth kinetics, total alkaloid content, and gene expression studies favored highly significant data (P < 0.05-0.01). Above treated hairy roots were also up-scaled in a 5-l stirred-tank bioreactor where a 40-day cycle yielded 8-fold increase in fresh root mass.


Synergistic effect of hen egg white lysozyme and lysosomotropic surfactants on cell viability and membrane permeability.
The interactions between two types of quaternary ammonium surfactants (N,N,N-trimethyl-2-(dodecanoyloxy)ethaneammonium bromide (DMM-11) and N,N,N-trimethyl-2-(dodecanoyloxy)propaneammonium bromide (DMPM-11)) and hen egg white lysozyme were studied through several techniques, including isothermal titration calorimetry (ITC), circular dichroism (CD) and fluorescence spectroscopy, and surface tension measurement. The average number of surfactants interacting with each molecule of lysozyme was calculated from the biophysical results. Moreover, the CD results showed that the conformation of lysozyme changed in the presence of DMM-11 and DMPM-11. The studies drew a detailed picture on the physicochemical nature of interactions between both surfactants and lysozyme. Both DMM-11 and DMPM-11, with and without lysozyme were studied against three target microorganisms, including Gram-negative (Escherichia coli) and Gram-positive (Enterococcus hirae and Enterococcus faecalis) bacteria. The results revealed a broad spectrum of antibacterial nature of surfactant/lysozyme complexes, as well as their effect on the membrane damage, hence providing the basis to further explore DMM-11 and DMPM-11 combined with lysozyme as possible antibacterial tools.


Development and application of an analytical approach to assess an antibody's potential for disulfide reduction.
Monoclonal antibody (mAb) interchain disulfide bond reduction has been observed in a recent large-scale clinical manufacturing operation. A massive reduction/precipitation at post-clarification steps has occurred. This note presents the development of a novel analytical approach to identify the "potential reduction"-a unique approach to predict the propensity of a monomeric-profiled mAb to be reduced in the post-harvest stage, such as harvest clarification and/or purification steps. The core of this new approach includes comparing the non-reducing capillary electrophoresis profiles of pre- and post-vacuum treated mAb in harvest cell culture fluid (HCCF). Using this approach, the potential reductions of two in-house mAbs in the unclarified and clarified cell culture harvest were assessed.


Isotachophoretic analysis of mandelic acid, phenylglyoxylic acid, hippuric acid and methylhippuric acid in urine after occupational exposure to styrene, toluene and/or xylene.
A simple, rapid and sensitive analytical method has been developed for the determination of phenylglyoxylic acid, mandelic acid, hippuric acid and methylhippuric acid; 0-, m- and p-methylhippuric acids are partly separated. These compounds are found as metabolites after occupational exposure to styrene, toluene and xylene. The method has been applied successfully to samples extracted from human urine by diethyl ether. The method can be used to accurately and simultaneously determine as little as 0.5 nmole of all of these acids in less than 20 min.


Expression of metallothionein in lung carcinoma: correlation with histological type and grade.
Over-expression of cellular metallothionein occurs frequently in human tumours but the underlying mechanism remains unknown. The aim of this study was to assess metallothionein expression in cases of lung carcinoma and to correlate it with histopathological parameters. Tumour tissue samples from 89 patients with lung carcinoma were immunostained by the streptavidin-biotin-peroxidase technique, using a monoclonal antibody against both metallothionein-1 and -2 isoforms. Positive matallothionein immunostaining was prominent in 44 out of 89 (49%) and negative in 45 out of 89 (51%) cases of lung carcinoma examined. Metallothionein positivity was prominent in 32 out of 43 (74%) cases of squamous cell lung carcinoma, and in 12 out of 35 (34%) cases of adenocarcinoma, while it was negative in all 11 cases of small-cell lung carcinoma examined, presenting a statistically significant difference between the different histological types. The intensity of metallothionein staining revealed a statistically significant difference between the squamous cell and adenocarcinoma cases examined. The pattern and extent of metallothionein staining in tumour cells and the expression of metallothionein in stromal cells were not correlated with histopathological parameters (type and grade) in metallothionein-positive cases of lung carcinoma examined. No association was found between metallothionein expression and lymph node status in the examined cases of lung carcinoma. Our findings indicate that expression of metallothionein was evident in squamous cell lung carcinoma and adenocarcinoma, but absent in small-cell lung carcinoma, supporting evidence for participation of this protein in the biological mechanisms underlying the carcinogenic evolution in the lung.


MicroRNA-22 inhibits tumor growth and metastasis in gastric cancer by directly targeting MMP14 and Snail.
MicroRNAs (miRNAs) deregulation is frequent in human gastric cancers (GCs), but the role of specific miRNAs involved in this disease remains elusive. MiR-22 was previously reported to act as tumor suppressors or oncogenes in diverse cancers. However, their accurate expression, function and mechanism in GC are largely unclear. Here, we found that the expression of miR-22 was significantly reduced in clinical GC tissues compared with paired adjacent normal tissues, and was significantly correlated with a more aggressive phenotype of GC in patients, and miR-22 low expression correlated with poor overall survival. The introduction of miR-22 markedly suppressed GC cell growth, migration and invasion, and inhibition of miR-22 promoted GC cell proliferation, migration and invasion in vitro. We further demonstrated that miR-22 acted as tumor suppressors through targeting extracellular matrix (ECM) remodeling member matrix metalloproteinase 14 (MMP14) and epithelial-to-mesenchymal transition (EMT) inducer Snail in GC. Moreover, ectopic expression of MMP14 or Snail restored inhibitory effects of miR-22 on cell migration and invasion in GC cells, and a negative relationship between the miR-22 expression and MMP14 or Snail mRNA levels was observed in GC. Finally, overexpression of miR-22 suppressed tumor growth, peritoneal dissemination and pulmonary metastasis in vivo. Taken together, we identified that miR-22 is a potent tumor suppressor in GC. MiR-22 downregulation promotes GC invasion and metastasis by upregulating MMP14 and Snail, and then inducing ECM remodeling and EMT. These findings provide a better understanding of the development and progression of GC and may be an important implication for future therapy of the GC.


An immunohistochemical study of a carcinoma of the parotid gland exhibiting both ductal and acinic cell differentiation.
A 76-year-old man underwent a subtotal parotidectomy for removal of a 3 cm. multicystic mass. The tumor was a salivary gland carcinoma, with both infiltrating and intraductal/intra-acinar components, exhibiting three histologic patterns: cribriform, papillary, and comedo-like. Immunohistochemical stain for keratin by the immunoperoxidase technique was strongly reactive in the vast majority of the tumor cells, indicating ductal differentiation of the tumor. Ultrastructural studies indicated primarily ductal differentiation of the tumor cells, with additional areas of acinous and myoepithelial differentiation.


FLUORESCENCE LIFETIME IMAGING OPHTHALMOSCOPY AS PREDICTOR OF LONG-TERM FUNCTIONAL OUTCOME IN MACULA-OFF RHEGMATOGENOUS RETINAL DETACHMENT.
To assess whether macular fluorescence lifetimes may serve as a predictor for long-term outcomes in macula-off rhegmatogenous retinal detachment. A single-center observational study was conducted. Patients with pseudophakic macula-off rhegmatogenous retinal detachment were included and evaluated 1 and 6 months after successful reattachment surgery. Fluorescence lifetime imaging ophthalmoscopy lifetimes in the central Early Treatment Diabetic Retinopathy Study grid subfield, in two distinct channels (short spectral channel and long spectral channel) were analyzed. Best-corrected visual acuity optical coherence tomography of the macula and fluorescence lifetimes were measured at month 1 and month 6. Nineteen patients were analyzed. Lifetimes of the previously detached retinas were prolonged compared with the healthy fellow eyes. Short lifetimes at month 1 were associated with better best-corrected visual acuity improvement (short spectral channel: r2 = 0.27, P < 0.05, long spectral channel: r2 = 0.23, P < 0.05) and with good final best-corrected visual acuity (short spectral channel: r2 = 0.43, P < 0.01, long spectral channel: r2 = 0.25, P < 0.05). Lifetimes were prolonged in some cases of outer retinal damage in optical coherence tomography scans. Fluorescence lifetime imaging ophthalmoscopy might serve as a prediction tool for functional recovery in pseudophakic macula-off rhegmatogenous retinal detachment. Retinal fluorescence lifetimes could give insight in molecular processes after rhegmatogenous retinal detachment.


The first benthic harmful dinoflagellate bloom in China: Morphology and toxicology of Prorocentrum concavum.
More frequent events and geographic expansion of benthic harmful algal blooms have been reported in recent years. An unexpected bloom of benthic P. concavum occurred in Xincun Bay, Hainan Island, the South China Sea was monitored in August 2018. Species identification, toxin analysis and toxicity test were conducted in the study. Quantitative study revealed that P. concavum had a high cell density on the surface of substrates and in water column. The bloom forming species was identified based on the morphology and phylogeny. Toxin analysis indicated that there were no detectable DSP toxins either in algae or in shellfish samples. The result of toxicity test revealed that the extracts of P. concavum caused the mortality of brine shrimp larvae (Artemia salina). The results from this study may provide more insight into the rising threats of harmful dinoflagellate blooms to marine benthic ecosystems.


Nociceptive responses to intrathecally administered substance P and somatostatin in diabetic mice.
Male ICR mice were rendered diabetic by i.v. injection of streptozotocin. The nociceptive behavioral responses to i.t. injection of somatostatin (SST) but nor substance P (SP) were attenuated in diabetic mice compared with that in non-diabetic mice. Spantide, a SP receptor antagonist, enhanced the nociceptive response induced by i.t. SST in diabetic mice. Pretreatment of mice with SP reduced the SST-induced nociceptive response in non-diabetic mice. These results suggest that a endogenous antinociceptive system may exist which links SP with SST-mediated nociceptive transmission in the spinal cord. Furthermore, this endogenous antinociceptive system may be enhanced in diabetic mice.


The Tannea Blackwelder species of Ecuador with description of new species (Coleoptera: Staphylinidae: Osoriinae).
Two new species of the genus Tannea Blackwelder from Ecuador, Tannea hermani n. sp. and Tannea bellavistae n. sp. are herein described, and a complete list of records of all Tannea species found in Ecuador is provided together with a key to the species. Additionally, the geographic distribution of the Tannea species of Ecuador and their altitudinal distribution are discussed.


Differential effects of cadmium on cytosolic and mitochondrial glutathione levels in the rat heart.
In rats fed a low-selenium diet, cytosolic GSH levels in the heart were significantly elevated when compared to rats fed a selenium-supplemented diet. Similar results were obtained in cardiac cytosol from rats treated with either parenteral or dietary Cd. Mitochondrial GSH was not affected by cadmium in selenium-deficient rats. However, significant elevations of mitochondrial GSH were seen in cadmium-treated rats fed either 0.1 or 0.5 ppm selenium. The alterations in cytosolic GSH levels correlate with reductions in GSH-Px activity reported earlier [(1987) Toxicol. Appl. Pharmacol. 87, in press]. However, levels of mitochondrial GSH did not correlate with GSH-Px activity.


The ratio and concentration of two monoterpenes mediate fecundity of the pinewood nematode and growth of its associated fungi.
The pinewood nematode (PWN) Bursaphelenchus xylophilus, vectored primarily by the sawyer beetle, Monochamus alternatus, is an important invasive pest and causal agent of pine wilt disease of Chinese Masson pine, Pinus massoniana. Previous work demonstrated that the ratios and concentrations of α-pinene:β-pinene differed between healthy trees and those trees containing blue-stain fungus (and M. alternatus pupae). However, the potential influence of the altered monoterpene ratios and concentrations on PWN and associated fungi remained unknown. Our current results show that low concentrations of the monoterpenes within petri dishes reduced PWN propagation, whereas the highest concentration of the monoterpenes increased PWN propagation. The propagation rate of PWN treated with the monoterpene ratio representative of blue-stain infected pine (α-pinene:β-pinene = 1:0.8, 137.6 mg/ml) was significantly higher than that (α-pinene:β-pinene = 1:0.1, 137.6 mg/ml) representative of healthy pines or those damaged by M. alternatus feeding, but without blue stain. Furthermore, inhibition of mycelial growth of associated fungi increased with the concentration of the monoterpenes α-pinene and β-pinene. Additionally, higher levels of β-pinene (α-pinene:β-pinene = 1:0.8) resulted in greater inhibition of the growth of the associated fungi Sporothrix sp.2 and Ophiostoma ips strains, but had no significant effects on the growth of Sporothrix sp.1, which is the best food resource for PWN. These results suggest that host monoterpenes generally reduce the reproduction of PWN. However, PWN utilizes high monoterpene concentrations and native blue-stain fungus Sporothrix sp.1 to improve its own propagation and overcome host resistance, which may provide clues to understanding the ecological mechanisms of PWN's successful invasion.


Development of Neuropathic Arthropathy With Entrectinib: Case Report.
TRK inhibition can lead to on-target neurologic adverse events. Two cases illustrate the development of neuropathic arthropathy as a side effect of entrectinib. After starting entrectinib, both patients developed foot pain, swelling, and sensory changes with magnetic resonance imaging revealing marrow edema. Providers should have a low threshold to investigate foot pain or gait abnormalities thoroughly with magnetic resonance imaging and referral to specialists to aid in the diagnosis of neuropathic arthropathy, with consideration to transition to an alternative agent.


Knockdown and mutation of Pou4f3 gene mutation promotes pyroptosis of cochleae in cisplatin-induced deafness mice by NLRP3/caspase-3/GSDME pathway.
To investigate the effect and mechanism of Pou4f3 gene mutation on pyroptosis in cochleae of cisplatin-induced deafness mice. Mice were intraperitoneally injected with cisplatin to construct an animal model of deafness, and sh-Pou4f3 and mutant vector were injected to alter gene expression. TUNEL staining was used to assess the apoptosis level of cochlear hair cells, ELISA was used to detect the secretion of inflammatory factors, and immunofluorescence and Western Blot were used to detect the expression of pyroptosis related factors. Cisplatin induced pyroptosis through NLRP3/Caspase-3/GSDME pathway and significantly down-regulated Pou4f3 level. Pou4f3 mutations promote cochlear hair cell pyroptosis by activating the NLRP3/Caspase-3/GSDME pathway. Knockdown of Pou4f3 can superimpose cisplatin treatment to induce pyroptosis of cochlear hair cells through NLRP3/Caspase-3/GSDME pathway. Pou4f3 gene mutation promotes pyroptosis of cochleae in cisplatin-induced deafness mice through NLRP3/Caspase-3/GSDME pathway.


Mathematical model to predict the need for neodymium: YAG capsulotomy based on posterior capsule opacification rate.
To evaluate a model to project the estimated time required before patients having primary phacoemulsification require neodymium:YAG (Nd:YAG) laser capsulotomy. Eleven private practices in the United States. Projections of time to capsulotomy were based on assessment of the early development of posterior capsule opacification (PCO) over time. The PCO data were collected during a clinical study to evaluate MDX-RA, an investigational immunotoxin designed to limit epithelial cell growth, preventing postsurgical PCO. From the PCO data, the estimated time to Nd:YAG capsulotomy in a placebo-treated group was compared with the actual time to capsulotomy in a cohort of patients from general practice who had had phacoemulsification. By 6 months, the mean Opacification Index in the MDX-RA group was significantly lower than that in the placebo group (P < .05) and it remained significantly lower at 12 (P < .001), 18 (P < .001), and 24 (P < .016) months. The rate of PCO in the MDX-RA group was approximately 6 times lower than that in the placebo group (P < .0004). Fifty-seven percent in the placebo group and 4% in the MDX-RA group were projected to require an Nd:YAG capsulotomy within 3 years of primary cataract surgery. Projected values for the placebo group were similar to actual values observed in the population-based cohort. This technique could be used to predict the need for Nd:YAG capsulotomy using early measurements of PCO.


Molecular modeling studies of structural properties of polyvinyl alcohol: a comparative study using INTERFACE force field.
Polyvinyl alcohol (PVA) is a material with a variety of applications in separation, biotechnology, and biomedicine. Using combined Monte Carlo and molecular dynamics techniques, we present an extensive comparative study of second- and third-generation force fields Universal, COMPASS, COMPASS II, PCFF, and the newly developed INTERFACE, as applied to this system. In particular, we show that an INTERFACE force field provides a possibility of composing a reliable atomistic model to reproduce density change of PVA matrix in a narrow temperature range (298-348 K) and calculate a thermal expansion coefficient with reasonable accuracy. Thus, the INTERFACE force field may be used to predict mechanical properties of the PVA system, being a scaffold for hydrogels, with much greater accuracy than latter approaches. Graphical abstract Molecular Dynamics and Monte Carlo studies indicate that it is possible to predict properties of the PVA in narrow temperature range by using the INTERFACE force field.


Nation-wide survey of screening practices to detect carriers of multi-drug resistant organisms upon admission to Swiss healthcare institutions.
As emergence and spread of multi-drug resistant organisms (MDRO) requires a standardized preventive approach, we aimed to evaluate current MDRO admission screening practices in Swiss hospitals and to identify potential barriers impeding their implementation. In early 2018, all Swiss public and private healthcare institutions providing inpatient care were contacted with a 34-item questionnaire to investigate current MDRO admission screening policies. Among 139 respondents representing 180 institutions (response rate, 79%), 83% (149) of institutions implemented MDRO admission screening, while 28% of private and 9% of public institutions did not perform any screening. Targeted high-risk screening included carbapenemase producers, extended-spectrum beta-lactamase producers and methicillin-resistant Staphylococcus aureus at the institutional level for respectively 78% (115), 81% (118) and 98% (145) of screening institutions. Vancomycin-resistant enterococci (44% of institutions), multi-resistant Acinetobacter baumanii (41%) and Pseudomonas aeruginosa (37%) were systematically searched only by a minority of screening institutions. A large diversity of risk factors for targeted screening and some heterogeneity in body sites screened were also observed. Admission-screening practices were mostly impeded by a difficulty to identify high-risk patients (44%) and non-compliance of healthcare workers (35%). Heterogeneous practices and gaps in small and privately-owned institutions, as well as a mismatch between current epidemiologic MDRO trends and screening practices were noticed. These results highlight the need for uniform national MDRO screening standards.


Filamentous phage as vector-mediated antibody delivery to the brain.
Early diagnosis of Alzheimer's disease is prevented by lack of means to visualize and target beta amyloid plaques in the brains of affected people. There are many methods of detecting amyloid plaques by staining postmortem brain tissue, but none are available for monitoring in living patients. We propose anti-beta amyloid antibodies as a highly specific probe to monitor amyloid plaque formation in living patients. Intranasal administration of filamentous phage as delivery vector of anti-beta amyloid antibody fragment into Alzheimer's APP transgenic mice enables in vivo targeting of beta amyloid plaques. The plaques were co-visualized both by thioflavin-S and fluorescent-labeled anti-phage antibodies in the olfactory bulb and the hippocampus region. The genetically engineered filamentous bacteriophage proved to be an efficient and nontoxic viral delivery vector to the brain, offering an obvious advantage over other mammalian vectors. The ability to image A beta deposits in vivo would arguably provide the most useful diagnostic and monitoring test for early diagnosis of Alzheimer's disease.


Indiana continent urinary reservoir.
Cecoileal reservoirs were created in 29 patients. Tunneled ureteral implantations along the tenia of the cecum provided the antireflux mechanism. Plication or tapering of the terminal ileal segment along with the ileocecal valve provided the continence mechanism. The tubular configuration of the cecum was disrupted with either an ileal or sigmoid patch, or it was re-configured in a Heineke-Mikulicz type of closure to avoid bolus (unit) contractions. Short-term followup examination with excretory urography showed no upper tract obstruction. X-rays of the pouch showed no reflux and interviews revealed satisfactory continence in 93 per cent of the patients.


Draft Genome Sequences of Exfoliative Toxin A-Producing Staphylococcus aureus Strains B-7772 and B-7777 (CC8/ST2993) and B-7774 (CC15/ST2126), Isolated in a Maternity Hospital in the Central Federal District of Russia.
Staphylococcus aureus clonal complex 8 (CC8) has not been associated with staphylococcal scalded-skin syndrome (SSSS) in newborns and exfoliative toxin genes. Here, we report the draft genome sequences of exfoliative toxin A-producing B-7772, B-7777 (both CC8), and B-7774 (CC15) strains associated with SSSS in newborns.


Synthesis of C-furanosides from a D-glucal-derived cyclopropane through a ring-expansion/ring-contraction sequence.
gem-Dibromocyclopropane 1, prepared from tri-O-benzyl-D-glucal, undergoes thermal and silver-promoted ring expansion in the presence of alcohols to give substituted oxepines. With further heating, ring contraction to highly substituted tetrahydrofurans follows. These represent C-furanosides, potentially useful as precursors to C-nucleosides and other carbohydrate mimics.


Infections in renal transplant patients: risk factors and infectious agents.
Infectious complications after renal transplantation (RT) are associated with significant morbidity. They continue to be the most frequent cause of mortality. We investigated the incidence of infections, the causative pathogens, and risk factors contributing to this complication during the first year. We included demographic and clinical data of the 124 patients who underwent RT in our hospital from December 2004 to June 2010. for statistical analysis. Fifty (40.3%) RT recipients developed 80 episodes of infection: urinary tract (n = 68; 85%), intraabdominal (n = 4; 5%), surgical wound (n = 3; 3.8%), or central venous catheter (n = 3; 4%). Eight (10%) were bacteremic. The most commonly isolated bacteria scene (76/80) was Escherichia coli (n = 43; 56.5%) followed by Klebsiella spp. (n = 10; 13.2%) and Pseudomonas spp. (n = 10; 13.2%). Cytomegalovirus infection was detected in 2 recipients; fungal and mycobacterial infections, in no case. It was noteworthy that 52.8% of E. coli and Klebsiella spp. produced extended-spectrum beta-lactamase. Ninety percent of infections developed within 6 months after transplantation. When we compared infected versus noninfected cases, the presence of a double J catheter was the most significant risk factor (P = .018; odds ratio [OR] = 0.234; 95% confidence interval [CI] = 0.070-0.781). In contrast to the initial years after the start of RT in our hospital the incidence of infection decreased over time together with a decrease number and durations of catheterization (P = .008; OR = 2.707; 95% CI = 1.292-5.672). Urinary tract infections were the predominant problem with most isolates resistant to extended-spectrum antibiotics. Therefore, invasive catheters and prophylactic antibiotics should not be used for longer than necessary and infection control measures implemented to decrease the incidence of infections and bacterial resistance.


Metabolism of phenothiazines: identification of N-oxygenated products by gas chromatography and mass spectrometry.
The solid inlet mass spectra of the N-oxide and N-oxide sulphoxide metabolites of promazine, chlorpromazine and methotrimeprazine have been determined at different temperatures and compared with the mass spectra of the main thermolytic product of the N-oxides and N-oxide sulphoxides obtained during combined gas chromatography-mass spectrometry. Diagnostic ions were produced by direct insertion of the compounds at ambient temperature into the mass spectrometer. These ions distinguish the N-oxides of arylalkyl tertiary amines from the N-oxygenated derivatives of primary and secondary arylalkylamines.


Evidence for the coupling of refill liquids content and new particle formation in electronic cigarette vapors.
The size and chemical content of particles in electronic cigarette vapors (e-vapors) dictate their fate in the human body. Understanding how particles in e-vapors are formed and their size is critical to identifying and mitigating the adverse consequences of vaping. Thermal decomposition and reactions of the refill liquid (e-liquid) components play a key role in new particles formation. Here we report the evolution of particle number concentration in e-vapors over time for variable mixtures of refill e-liquids and operating conditions. Particle with aerodynamic diameter < 300 nm accounted for up to 17% (or 780 μg/m3) of e-vapors particles. Two events of increasing particle number concentration were observed, 2-3 s after puff completion and a second 4-5 s later. The intensity of each event varied by the abundance of propylene glycol, glycerol, and flavorings in e-liquids. Propylene glycol and glycerol were associated with the first event. Flavorings containing aromatic and aliphatic unsaturated functional groups were strongly associated with the second event and to a lesser extent with the first one. The results indicate that particles in e-vapors may be formed through the heteromolecular condensation of propylene glycol, glycerol, and flavorings, including both parent chemicals and/or their thermal decomposition products.


CFTR modulators increase risk of acute pancreatitis in pancreatic insufficient patients with cystic fibrosis.
Patients with pancreatic insufficient cystic fibrosis rarely develop acute pancreatitis due to insufficient acinar reserve. We describe a series of five patients under the age of 18 (range 8-16 years) with pancreatic insufficient cystic fibrosis who developed a phenotype in keeping with acute pancreatitis following initiation of CFTR modulator therapy. This occurred at a median of 30 months following CFTR modulator initiation. 3/5 of these patients also developed pancreatic sufficiency or at least an intermediary pancreas status, indicated by fecal elastases above 100 μg/g. This series highlights a mostly unrecognized potential side effect of this therapy as well as the potential of CFTR modulator therapies to improve exocrine pancreatic function, even in adolescent patients.


Quorum quenchers affect the virulence regulation of non-mucoid, mucoid and heavily mucoid biofilms co-cultured on cell lines.
Biofilm formation conferring pathogenicity is a survival strategy for Pseudomonas aeruginosa. P. aeruginosa's virulence may differ due to differences in host-microbe interactions and the growth environment. The epithelial cell line within the respiratory system and the keratinocytes on the skin form the first physical barrier of defence. P. aeruginosa spp. biofilm formation and virulence factor secretion with and without quorum quenching (QQ) treatment was studied in co-culture using A549 and HaCaT cell lines; pyocyanin and rhamnolipid productions and elastolytic activity as virulence factors were quantified by independent assays. Biofilm formation was evaluated under dynamic conditions by quantifying total carbohydrates, alginate, proteins and eDNA. A sandwich ELISA was performed to study IL-8 secretion by the epithelial cells. The difference in gene expression of the quorum sensing (QS) and virulence factors between strains during individual and combination treatments was analysed by qPCR. Combination treatment by farnesol and tyrosol was more effective against P. aeruginosa biofilms when grown in co-cultures. The strain RBHi was found to be 3 to 4 times more virulent compared to PAO1 and NCTC 10,662, respectively, and combination treatment was more effective against RBHi strain when grown in co-culture with A549 cell line. The addition of quorum quenchers (QQs) individually and in combination reduced IL-8 secretion by A549 cells. Relative mRNA expression showed upregulation of the QS genes and virulence factors. Co-culture of P. aeruginosa and HaCaT cell line showed a general decrease in gene expression, especially in the case of P. aeruginosa RBHi when treated with farnesol and tyrosol combination.Key points• Differentiating the interactions of biofilm formed by different phenotypes of P. aeruginosa, NCTC 10,662 (non-mucoid), PAO1 (semi mucoid) and RBHi (heavily mucoid).• Biofilm formed by these P. aeruginosa strains on two commonly afflicted tissues represented by A549 (lung) and HaCaT (skin) cell lines.• Anti-biofilm/anti-virulence roles of quorum quenchers, tyrosol and farnesol in co-cultures.


Toddlers' and Preschoolers' Experience in Family Day Care: Age Differences and Behavioral Correlates.
One hundred and twelve children, 56 toddlers and 56 preschoolers, were observed in their family child care settings to determine whether toddlers cared for in settings that also included preschoolers were, relative to the preschoolers, receiving more or less high-quality care and/or whether their functioning at child care appeared to be more or less dependent on aspects of the care providers' interactions with the children. Quality of care was analyzed along two dimensions: Sensitive/Supportive Care and Structured Care. Four indices of child functioning at child care were examined: integration in social activities, attention, positive mood, and angry/aggressive behavior. Results indicate that toddlers received less sensitive, supportive care than preschoolers in these mixed-age settings and toddlers were less socially integrated and less engaged in activities in the child care setting. Preschoolers displayed increased levels of angry/aggressive behavior relative to toddlers. In addition, associations of care provider behaviors and child functioning were larger for toddlers than preschoolers, suggesting that toddlers were more dependent on caregiver support for more successful functioning in these family child care settings. For both toddlers and preschoolers, care provider behavior and child functioning was generally poorer in settings with more children.


MSPD sample preparation approach for reversed-phase liquid chromatographic analysis of pesticide residues in stem of coconut palm.
A method was developed using matrix solid-phase dispersion, together with liquid chromatography with ultraviolet diode array detector for determination of carbofuran, difenoconazole, β-cyfluthrin, spirodiclofen and thiophanate-methyl in stem of coconut palm. The best results were obtained using 2.0 g of stem, 1.6 g of Florisil as sorbent and cyclohexane:acetone mixture (4:1). The method was validated using stem samples spiked with pesticides at four concentration levels (0.05-2.0 μg/g). Average recoveries ranged from 70 % to 114.3 %, with relative standard deviations between 1.2 % and 19.2 %. Detection and quantification limits were in the ranges 0.02-0.03 and 0.05-0.1 μg/g, respectively.


[Evaluation of a screening strategy after occurrence of two simultaneous contaminating tuberculosis cases in a pediatric oncology department].
A medical staff and an administrative staff of our paediatric oncology department have contracted a pulmonary tuberculosis. This is a rare situation and the management of this infection threat in a paediatric oncology department is not clearly defined. Recommendations tell we must treat all patients. Nevertheless, antituberculosis agent expose to increased toxic effects and immunocompromised patients have an increased risk of experiencing progression of latent mycobacterium tuberculosis infection to active tuberculosis disease. This study aims at the evaluation of a screening and a treatment strategy adapted for a paediatric oncology department. From April 2004 to April 2005, 80 children with a solid tumour were screened for tuberculosis according to a screening and treatment protocol established by a multidisciplinary committee. Two risk groups were defined according to age and immunodepression status. The "high risk" group is composed of less than 2 years old children and children who underwent an haematological peripheral stem cell transplantation. All other children were included in the "low risk" group. The screening was based on clinical, biological and radiological data performed three times spaced out by 2 or 3 months. At the end of each part of screening, the multidisciplinary committee analyzed the results and discussed the utility of an antituberculosis treatment. 80 children (31 boys and 49 girl) with a median age of 7,3 years (0,3-24) participated to the screening. Sixty children were still undergoing anticancer treatments. Twenty belonged to the high risk group. The complete screening was performed in 32% of the patients. Three antituberculosis' treatment were initiated: 2 for prophylaxis purpose and 1 for a tuberculosis prime-infection. A child had an additional check-up because of an abnormal chest X-ray. Our management strategy allowed us to treat significantly less patients when compared to national guidelines (3 vs 80 test Chi-2 p<0.001). No side effects of antituberculosis agents were noted. No tuberculosis has been observed in our population 28 months after the completion of the treatment. The proposed screening allowed us to treat a minimum of children and thus, to reduce the potential toxicity induced by antituberculosis' treatments.


Non-Gaussian models of diffusion weighted imaging for detection and characterization of prostate cancer: a systematic review and meta-analysis.
The importance of Diffusion Weighted Imaging (DWI) in prostate cancer (PCa) diagnosis have been widely handled in literature. In the last decade, due to the mono-exponential model limitations, several studies investigated non-Gaussian DWI models and their utility in PCa diagnosis. Since their results were often inconsistent and conflicting, we performed a systematic review of studies from 2012 examining the most commonly used Non-Gaussian DWI models for PCa detection and characterization. A meta-analysis was conducted to assess the ability of each Non-Gaussian model to detect PCa lesions and distinguish between low and intermediate/high grade lesions. Weighted mean differences and 95% confidence intervals were calculated and the heterogeneity was estimated using the I2 statistic. 29 studies were selected for the systematic review, whose results showed inconsistence and an unclear idea about the actual usefulness and the added value of the Non-Gaussian model parameters. 12 studies were considered in the meta-analyses, which showed statistical significance for several non-Gaussian parameters for PCa detection, and to a lesser extent for PCa characterization. Our findings showed that Non-Gaussian model parameters may potentially play a role in the detection and characterization of PCa but further studies are required to identify a standardized DWI acquisition protocol for PCa diagnosis.


Functional in vivo verification in E. coli of promoter activities from the rDNA/tDNA(Val)(GAC) leader region of Zea mays chloroplasts.
Restriction fragments containing upstream sequences of the rRNA operon from Zea mays chloroplasts were tested for promoter activity in vivo by insertion into an E. coli promoter-probe vector. The expression of this vector's reporter gene, which codes for alkaline phosphatase, was stimulated more than 1,500-fold upon linkage with the chloroplast rRNA promoter. Site specific mutagenesis of the invariant T of the -10 sequence of this promoter reduced the expression of the reporter gene to 2% of the wild type. This indicates that the chloroplast rRNA promoter, which directs transcriptional initiation 117 bp upstream of the 16S rRNA gene, is also active in the bacterial system. A restriction fragment further upstream containing the gene for tRNA(Val) (GAC) also showed strong promoter activity (29% as compared with the rRNA promoter). This promoter activity probably reflects the chloroplast promoter directing the synthesis of the tRNA(Val) (GAC) primary transcript. Surprisingly, this restriction fragment also displayed promoter activity (13% compared with the rRNA promoter) in reverse orientation.


Mitogenic and polyclonal B cell activation activities of synthetic lipid A analogues.
The activation of murine B cells and macrophages by synthetic lipid A analogues, solubilized with triethylamine and complexed with bovine serum albumin, were investigated in vitro. The analogues used are nonphosphorylated, C-1 or C-4' monophosphorylated and C-1,4' diphosphorylated derivatives of beta-1,6-linked D-glucosamine disaccharide possessing both ester- and amide-bound fatty acid substituents. They were divided into 4 groups, A, B, C and D in terms of the fatty acid substitution. Ester- and amine-bound fatty acids of the analogues are both tetradecanoic acids (C14) in group A, C14 and (R)-3-hydroxytetradecanoic acids (C14-OH) in group B, both C14-OH in group C and C14-OH and (R)-3-tetradecanoyloxytetradecanoic acids in group D. Mitogenic activity was exhibited in spleen cells from C3H/HeN mice by the C-1 monophosphorylated analogues in groups A and B, and by the C-4' monophosphorylated analogues in groups B, C and D, but not in cells from C3H/HeJ mice. Nonphosphorylated analogues in groups A, B and C, and C-4' monophosphorylated analogue in group A showed negative mitogenic activity. Only the nonphosphorylated analogue in group D exhibited mitogenic activity in spleen cells from C3H/HeJ mice as well as those from C3H/HeN mice. None of the other analogues exhibited the activity in C3H/HeJ spleen cells. Polyclonal B cell activation activity was exhibited by the C-1 monophosphorylated analogues in groups A and B, and by the C-4' monophosphorylated analogues in groups C and D. Nonphosphorylated analogues in all groups and C-4' monophosphorylated analogues in groups A and B showed negative PBA activity. None of the analogues tested could induce any cytostatic macrophages from thioglycollate-elicited peritoneal macrophages.


[Method of determining the pteridine sensitivity of parasitic bacteria of the genus Bdellovibrio].
The method of determination of parasitic bacteria Bdellovibrio sensitivity to pteridine has been described. The method suggested can be used in research work for diagnostics of Bdellovibrio genus bacteria.


Study on the Role of Polymorphisms of the SOX-6 and MYB Genes and Fetal Hemoglobin Levels in Sicilian Patients with β-Thalassemia and Sickle Cell Disease.
The hemoglobinopathies, as β-thalassemia (β-thal) and sickle cell disease, are the most common hereditary hemolytic anemias. The increase of fetal hemoglobin (Hb F) levels can ameliorate the symptoms of hemoglobinopathies. There are several transcription factors such as MYB and SOX-6, which are involved in the regulation of Hb F. There are not enough studies investigating the association between single nucleotide polymorphisms (SNPs) of the SOX-6 and MYB genes and the variation of Hb F levels in patients affected by sickle cell disease and β-thal. We therefore decided to analyze the role of four missense variants of MYB and SOX-6 genes in the regulation of Hb F levels. In order to do so, we examinated 30 Sicilian patients affected by sickle cell disease and β-thal, to understand if these variants could also have an influence in our populations. Comparing two groups of patients with low and high levels of Hb F, we found no significant differences in the genetic distribution and allelic frequency of MYB and SOX-6 gene polymorphisms. We also created and compared a 'high producer' and 'low producer' genotype with different genes achieving the same result of no significant difference. Our results may be due either to the fact that the association between these genes and the regulation of Hb F levels are influenced by environmental history and population genetics, or to the small number of samples being analyzed.


Effects of residual S-metolachlor in soil on the phyllosphere microbial communities of wheat (Triticum aestivum L.).
S-metolachlor (S-ME) is a widely used chiral herbicide that can cause potential ecological risks via long-term usage. In this work, we chose a model plant, wheat, as the test material to determine the effects of applying 10 mg/kg S-ME to soil on its fresh weight, chlorophyll and malondialdehyde (MDA) content, and superoxide dismutase (SOD) activity and the diversity and structural composition of the phyllosphere microorganisms after 7 and 14 days of exposure. Our work showed that this concentration of residual S-ME in soil only slightly decreased plant biomass and had little effect on lipid peroxidation, the antioxidant enzyme system and chlorophyll content. Interestingly, although the test concentration of S-ME did not exert strong inhibitory effects on the physiological activities of wheat, it decreased the diversity of phyllosphere microbial communities and changed their structure, indicating that microorganisms were more sensitive stress indicators. S-ME reduced the colonization by some beneficial bacteria related to plant nitrogen fixation among the phyllosphere microorganisms, which influenced the growth and yield of wheat because these bacteria contribute to plant fitness. In addition, S-ME affected the association between the host and the composition of the phyllosphere microbial communities under different growth conditions. Our work provides insights into the ecological implications of the effects of herbicides on the phyllosphere microbiome.


ESWT for tendinopathy: technology and clinical implications.
The general consensus that tendinopathy, at least in the chronic stage, is mainly a degenerative condition and inflammation plays a minor role has led to a shift from treatments that target inflammation towards treatment options that promote regeneration. One of these treatments is extracorporeal shockwave therapy (ESWT), a physical therapy modality that uses pressure waves to treat tendinopathy. This review was undertaken to give an overview of the literature concerning this treatment, and special attention is given to the differences between focused and radial ESWT. A narrative description of wave characteristics, generation methods and in vitro effects of ESWT is given. The literature on ESWT as a treatment for one common tendinopathy, patellar tendinopathy, was systematically reviewed. Waves that are generated for focused and radial ESWT have very different physical characteristics. It is unclear how these characteristics are related to clinical effectiveness. Studies into the biological effects of ESWT have mainly used focused shockwave therapy, showing a number of effects of shockwaves on biological tissue. The systematic review of studies into the clinical effects of ESWT for patellar tendinopathy showed conflicting evidence for its effectiveness. Physical characteristics of focused and radial waves differ substantially, but effect on clinical effectiveness is unclear. Whereas in vitro studies often show the effects of ESWT on tendon tissue, results of clinical studies are inconsistent. Based on the review of the literature, suggestions are given for the use of ESWT in clinical practice regarding timing and treatment parameters.


Distal shower embolization during directional coronary atherectomy and stenting for diffuse stenosis of right coronary artery: current limitations of intravascular ultrasonography for evaluating fragile plaque.
A 74 year-old male with old anterior and inferior myocardial infarctions was treated with staged percutaneous coronary intervention. A chronic total occlusion of the middle segment of the left anterior descending branch was successfully stented during the first stage, and during the second stage, preprocedural intravascular ultrasonography (IVUS) revealed that the proximal segment of the right coronary artery was diffusely stenosed by mixed plaque. Directional coronary atherectomy under IVUS guidance was performed, but coronary slow flow appeared during the procedure. After successfully bailing out with intracoronary nicorandil, percutaneous thrombectomy and manual blood pumping, 2 coronary stents were implanted to fully cover the lesion. Quite contrary to expectation, the no-reflow phenomenon appeared just after post-dilatation and a repeat of the same maneuver could not completely recover coronary flow. Plaque fragility could not be predicted from the IVUS examination.


Genotyping Dickeya dianthicola Causing Potato Blackleg and Soft Rot Outbreak Associated With Inoculum Geography in the United States.
An outbreak of blackleg and soft rot of potato, caused primarily by the bacterial pathogen Dickeya dianthicola, has resulted in significant economic losses in the northeastern United States since 2015. The spread of this seedborne disease is highly associated with seed distribution; therefore, the pathogen likely spread with seed tubers. To describe the blackleg epidemic and track inoculum origins, a total of 1,183 potato samples were collected from 11 states associated with blackleg outbreak from 2015 to 2019. Of these samples, 39.8% tested positive for D. dianthicola. Seventeen isolates of D. dianthicola were recovered from these samples and the genetic diversity of these isolates was examined. Fingerprinting with BOX-A1R-based repetitive extragenic palindromic PCR and phylogenetic analysis based on sequences of the 16S rRNA and gapA genes indicated that D. dianthicola isolates were divided into three genotypes, denoted types I, II, and III. Ninety-five percent of samples from Maine were type I. Type II was found in Maine only in 2015 and 2018. Type II was present throughout the 5 years in some states at a lower percentage than type I. Type III was found in Pennsylvania, New Jersey, and Massachusetts, but not in Maine. Therefore, type I appears to be associated with Maine, but type II appeared to be distributed throughout the northeastern United States. The type II and rarer type III strains were closer to the D. dianthicola type strain isolated from the United Kingdom. This work provides evidence that the outbreak of blackleg of potato in the northeastern United States was caused by multiple strains of D. dianthicola. The geographic origins of these strains remain unknown.


Repair of a bifid nose combined with a cleft of the primary palate in a 1-year-old dog.
To report surgical repair of a bifid nose combined with a cleft of the primary palate. Clinical report. A 1-year-old, male castrated Springer spaniel dog. With the dog in sternal recumbency, an extraoral (dorsal) approach to the nose was performed, and after surgical margins were outlined, a Y-shaped skin incision was made to remove redundant tissue and expose the bifid nasal cartilages. The cartilages were opposed and sutured together and the skin closed in 2 layers. The dog was repositioned in dorsal recumbency, and the 2 maxillary first incisor teeth were extracted. After tangential incision and undermining of the cleft defect, the mucosa was sutured in 1 layer. Healing was uneventful and there was an immediate return to normal function. At 2 weeks, 3 and 6 months function was excellent without further clinical signs. Bifid nose associated with a cleft of the primary palate can be surgically corrected.


Synthesis and aromatase inhibition by potential metabolites of exemestane (6-methylenandrosta-1,4-diene-3,17-dione).
Exemestane (6-methylenandrosta-1,4-diene-3,17-dione; FCE 24304) is an orally active irreversible aromatase inhibitor which is in phase II clinical evaluation for the potential therapy of postmenopausal breast cancer. A series of exemestane analogs, with modifications at the 6-methylene group and with additional reduction at the 17-keto group, were synthesized as potential metabolites and tested in vitro for their effect on human placental aromatase. All these new analogs were found to be less potent in inhibiting aromatase than exemestane. The most effective compound was the 17 beta-hydroxy-derivative (compound 2), which is 2.6-fold less potent than exemestane [50% inhibitory concentration (IC50) 69 and 27 nM, respectively]. The various C-6 modified derivatives of the 17-oxo series were found to inhibit the aromatase enzyme in the following descending order: 6-methylene (exemestane) > 6-spirooxirane (6) > 6 beta-hydroxymethyl (11) > 6-hydroxymethyl (7) > 6 beta-carboxy (13), showing IC50 values of 27, 206, 295, 2,300, and 7,200 nM, respectively. The 17 beta-hydroxy analogs of some of the above mentioned compounds were also synthesized (3,4,12) and found to be 3-8-fold less potent than the corresponding 17-keto analogs.


Trade-off among antiherbivore defences in a south american blackberry (Rubus bogotensis).
The idea of trade-offs among antiherbivore defences in plants is examined using data from a South American blackberry (Rubus bogotensis). Two distinct morphs of R. bogotensis, one with glandular trichomes and one without, were compared with respect to leaf toughness, number of prickles and prickle length. The two morphs were sympatric and grew under similar environmental conditions. The morph lacking trichomes had significantly tougher leaves and also tended to have more and longer prickles. Bioassay showed that Ithomiid larvae fed to a lesser extent on tough leaves than on more tender ones. Correlations between antiherbivore defences within each phenotype revealed three significant or almost significant negative relationships. The comparisons support the hypothesis that trade-offs exist among antiherbivore defences.


Plasma glutathione S-transferase measurements in patients with alcoholic cirrhosis.
Hepatocellular damage has been assessed in 54 patients with biopsy proven alcoholic cirrhosis by measuring the activity of aspartate aminotransferase (AST) and the concentrations of glutathione S-transferase B1B1 (GST B1B1) and B2B2 (GST B2B2) in serum. The levels of AST, GST B1B1, or GST B2B2 were abnormal in 28, 28 and 17 patients respectively but abnormalities in AST and GST measurements appeared to identify different populations of patients.


Diabetes Prevention Program community outreach: perspectives on lifestyle training and translation.
The gap between what is known from clinical efficacy research and the systematic community translation of diabetes prevention programs is narrowing. During the past 5 years, numerous randomized and nonrandomized dissemination studies have evaluated the modified delivery of structured Diabetes Prevention Program (DPP) interventions in diverse real-world settings. Programs of sufficient dose and duration, implemented with fidelity, have reported weight losses in the range of 4%-7% with associated improvements in cardiometabolic risk factors at 6 and 12 months from baseline. The current article describes some of the experiences and perspectives of a team of University of Pittsburgh researchers as they have engaged in these efforts.


Single-scan volume-selective spectral editing by homonuclear polarization transfer.
An RF and field gradient pulse sequence is presented permitting the single-scan volume-selective spectral editing of the signals of coupled spins. The attribute "single scan" means that the editing of resonance lines is performed in each accumulation or phase cycling scan rather than by subtraction of independently acquired signals. As an example the methyl line of lactate has been investigated. The method is considered to be a useful tool in biomedical research using lactate as an ischemic parameter. The basic principles applied are the VOSY pulse scheme for volume selection and the INEPT sequence for homonuclear polarization transfer from the CH to the CH3 groups. As an intrinsically single-scan editing method ("homonuclear single-scan VOSING procedure"), the pulse train is expected to provide results that are more reliable than the two-scan subtraction methods previously reported.


Effect of Rebamipide on the Premalignant Progression of Chronic Gastritis: A Randomized Controlled Study.
Chronic gastritis frequently progresses into precancerous intestinal metaplasia and intraepithelial neoplasia lesions. Rebamipide is a free radical scavenger and we assessed its efficacy on clinical symptoms, gastric mucosal lesions, pathologic grade, and immunohistochemistry in chronic gastritis patients. 178 eligible patients were randomized into treatment and control groups. Both groups followed an optimized lifestyle for 26 weeks, but the treatment group was additionally medicated with rebamipide 0.1 g three times per day. Upper gastrointestinal endoscopy was performed in all patients to evaluate the severity of gastritis by the Modified Lanza Scoring (MLS) and histological changes were evaluated by the Updated Sydney System Score (USSS). Gastric mucosa immunohistochemistry in the treatment group was performed using the intestinal metaplasia markers caudal type homeobox transcription factor 2 (CDX2) and trefoil factor 3 (TFF3) detection. There were significant outcome differences between the treatment and control groups regarding the clinical symptom scores (2.62 ± 1.86 vs. 1.55 ± 1.61, P = 0.0001), gastric mucosal lesion scores (0.57 ± 1.05 vs. 0.16 ± 0.90, P = 0.002), and inflammation (P < 0.05). Only in the treated patients were the rates of intestinal metaplasia (P = 0.017 vs. P = 0.123) and low-grade intraepithelial neoplasia (P = 0.005 vs. P = 0.226) significantly reduced after 26 weeks. The percentages of CDX2 (31.5 vs. 15.7%, P = 0.021) and TFF3 (44.9 vs. 25.8%, P = 0.012) expressing gastric mucosa cells were significantly lower after rebamipide medication than pre-treatment values. Rebamipide improved the clinical symptoms, gastric mucosal lesions, and pathological grades of chronic gastritis patients and decreased the expression rates of CDX2 and TFF3 in gastric cells.


Temporal trends and impact of willingness to accept organs from donors with hepatitis C virus.
Direct-acting antivirals (DAA) transformed hepatitis C virus (HCV) treatment in 2014; however, their impact on transplant candidates' willingness to accept (CWTA) organs from HCV+ donors remains uncertain. We retrospectively studied Organ Procurement and Transplantation Network data from 2008 to 2019, investigating CWTA different organs from HCV+ donors over time, using segmented multivariable logistic regression, and how that influenced wait-time and deceased-donor transplantation (DDTx) probability, using multivariable logistic or linear regression. We found that DAA availability was associated with a marked increase in CWTA in all organs from HCV+ donors except intestine. By December 2020, 40% of kidney, 33% of kidney-pancreas, 42% of pancreas, over 50% of liver, heart, lung, heart-lung, and 9% of intestine candidates waitlisted were CWTA an organ from HCV+ donors. Compared with pre-DAA, yearly CWTA kidney from HCV+ donors increased post-DAA 1.78 1.811.83 -fold, kidney-pancreas 2 .52 2.78 3.07 -fold, pancreas 3.15 3.69 4.43 -fold, liver 1.53 1.541.56 -fold, heart 1 .92 2.02 .08 -fold, and lung 2.00 2.12 .20 -fold. CWTA kidney and liver from HCV+ donors significantly increased DDTx probability post-DAA (1.98 2.042.1 -fold and 1.24 1.291.33 -fold, respectively) and shortened kidney candidates' wait-time78 90101 days (Mean with 95% CI). CWTA organs from HCV+ donors rose significantly with DAA availability, benefitting kidney and liver candidates with increased DDTx rates and shortened kidney candidates' wait time. Further long-term outcomes investigation and standardized organ from HCV+ donors' education could improve both provider and patient acceptance and utilization.


Ethanol increases matrix metalloproteinase-12 expression via NADPH oxidase-dependent ROS production in macrophages.
Matrix metalloproteinase-12 (MMP-12), an enzyme responsible for degradation of extracellular matrix, plays an important role in the progression of various diseases, including inflammation and fibrosis. Although most of those are pathogenic conditions induced by ethanol ingestion, the effect of ethanol on MMP-12 has not been explored. In the present study, we investigated the effect of ethanol on MMP-12 expression and its potential mechanisms in macrophages. Here, we demonstrated that ethanol treatment increased MMP-12 expression in primary murine peritoneal macrophages and RAW 264.7 macrophages at both mRNA and protein levels. Ethanol treatment also significantly increased the activity of nicotinamide adenine dinucleotide (NADPH) oxidase and the expression of NADPH oxidase-2 (Nox2). Pretreatment with an anti-oxidant (N-acetyl cysteine) or a selective inhibitor of NADPH oxidase (diphenyleneiodonium chloride (DPI)) prevented ethanol-induced MMP-12 expression. Furthermore, knockdown of Nox2 by small interfering RNA (siRNA) prevented ethanol-induced ROS production and MMP-12 expression in RAW 264.7 macrophages, indicating a critical role for Nox2 in ethanol-induced intracellular ROS production and MMP-12 expression in macrophages. We also showed that ethanol-induced Nox2 expression was suppressed by transient transfection with dominant negative IκB-α plasmid or pretreatment with Bay 11-7082, a selective inhibitor of NF-κB, in RAW 264.7 macrophages. In addition, ethanol-induced Nox2 expression was also attenuated by treatment with a selective inhibitor of p38 MAPK, suggesting involvement of p38 MAPK/NF-κB pathway in ethanol-induced Nox2 expression. Taken together, these results demonstrate that ethanol treatment elicited increase in MMP-12 expression via increase in ROS production derived from Nox2 in macrophages.


A role for prostaglandins in estrogen growth regulation.
Several lines of evidence support the theory that estrogen increases prostaglandin F2 alpha (PGF2 alpha) receptor number and amplifies the intracellular response to the uterine mitogen, PGF2 alpha, in primary cultures of rabbit endometrium. It is proposed here that estrogen induction of growth in target tissues may be mediated through prostaglandins. Determination of prostaglandin receptor status and prostaglandin levels in estrogen dependent tumors could be of importance as diagnostic tools. The use of prostaglandin synthesis inhibitors and/or antagonists in the therapy of these tumors should be considered.


The effect of breakfast on social behavior and brain amine metabolism in vervet monkeys.
Two groups of vervet monkeys were fed, on alternate days, either before or after a morning observation period. This enabled us to determine changes in behavior when the animals were fed a nutritionally balanced breakfast of monkey chow. Feeding did not alter the proportion of behaviors that were social or non-social, but had a marked effect on individual behaviors. Feeding increased active behaviors among the adult animals except for the vervets who were lowest in the social hierarchy in each cage. For some of the individual behaviors that were altered by feeding, the changes were most marked early on in the observation period, when the animals were still feeding. Other behavioral changes were seen only later in the observation period, a time course consistent with a food-mediated change in brain biochemistry. A parallel biochemical experiment showed that feeding decreased the levels of tryptophan and 5-hydroxyindoleacetic acid in the CSF. Our data indicate that feeding can influence both brain biochemistry and behavior. The behavioral changes may be influenced by social and psychological factors as well as changes in brain biochemistry.


[Ultra-structural study of the lateral portion of the auditory sensorial organ using a decalcification-free method].
To explain the development of a new personal technique to study the spiral ligament and stria vascularis in Guinea pig cochleae by obtaining sample tissue without decalcification and to assess its validity for electron microscopy analysis. Samples were taken from five female Guinea pigs weighing 200-250 g and were fixed in glutaraldehyde and paraformaldehyde for analysis of the spiral ligament and stria vascularis ultrastructure by transmission electron microscopy. All of the ultrastructure components in the spiral ligament and stria vascularis could be examined without the need for decalcification. Our method to obtain and analyze samples of cochlea side wall is valid, easy and faster.


In vitro skin toxicity of CuO and ZnO nanoparticles: Application in the safety assessment of antimicrobial coated textiles.
In the context of nosocomial infections, there is an urgent need to develop efficient nanomaterials (NMs) with antibacterial properties for the prevention of infection diseases. Metal oxide nanoparticles (MeO-NPs) are promising candidates for the development of new antibacterial textiles. However, the direct exposure to MeO-NPs and MeO-coated NMs through skin contact could constitute a severe hazard for human health. In this work, the toxicity of copper and zinc oxide (CuO, ZnO) NPs antimicrobial-coated textiles was assessed on an in vitro reconstructed 3D model of epidermis. Thus, MeO-NPs and extracts from MeO-coated NMs were tested on EpiDerm™ skin model according to OECD TG 431 (Corrosion Test) and 439 (Irritation Test), respectively. Skin surface fluids composition is a crucial aspect to be considered in the development of NMs that have to encounter this tissue. So, for the irritation test, coated textiles were extracted in artificial sweat solutions at pH 4.7 and 6.5. Skin tissue viability, pro-inflammatory interleukin-8 secretion and morphological alteration of intermediate and actin filaments of keratinocytes were evaluated after 18 h exposure to extracts from CuO- and ZnO-coated textiles. Analysis of extracts at the two pH conditions indicated that released ions and not NPs are involved in promoting adverse effects on epidermis. Since Cu2+ and Zn2+ ions are known to penetrate epidermis, Balb/3 T3 cells were used as model of dermis. Fibroblasts viability was investigated after the exposure to trans-epidermis permeated ions, collected from EpiDerm™ basal supernatants, and to extracts, as representative of a direct interaction of ions with dermis cells by wounded skin. From our data we can conclude that: 1) skin surface fluids composition is a key parameter for the stability of NPs-coated textiles; 2) MeO ions released from coated textiles can deeply affect the epidermal tissue and the underlying dermal cells upon trans-epidermal permeation; 3) skin barrier integrity is a fundamental prerequisite that should be taken into account during the assessment of NMs safety by direct contact exposure.


A Novel Class of ER Membrane Proteins Regulates ER-Associated Endosome Fission.
Endoplasmic reticulum (ER) membrane contact sites (MCSs) mark positions where endosomes undergo fission for cargo sorting. To define the role of ER at this unique MCS, we targeted a promiscuous biotin ligase to cargo-sorting domains on endosome buds. This strategy identified the ER membrane protein TMCC1, a member of a conserved protein family. TMCC1 concentrates at the ER-endosome MCSs that are spatially and temporally linked to endosome fission. When TMCC1 is depleted, endosome morphology is normal, buds still form, but ER-associated bud fission and subsequent cargo sorting to the Golgi are impaired. We find that the endosome-localized actin regulator Coronin 1C is required for ER-associated fission of actin-dependent cargo-sorting domains. Coronin 1C is recruited to endosome buds independently of TMCC1, while TMCC1/ER recruitment requires Coronin 1C. This link between TMCC1 and Coronin 1C suggests that the timing of TMCC1-dependent ER recruitment is tightly regulated to occur after cargo has been properly sequestered into the bud.


Gender differences in acute coronary syndromes patterns during the COVID-19 outbreak.
Mortality from acute coronary syndromes (ACS) is strictly related to early management. As female patients usually experience longer delays before diagnosis and treatment, we assessed whether women were more affected by the dramatic drop in hospital admissions for ACS during the Covid-19 pandemic. We performed a retrospective analysis of clinical and angiographic characteristics of consecutive patients who were admitted for ACS at 15 hospitals in Northern Italy comparing men and women data. The study period was defined as the time between the first confirmed case of Covid-19 in Italy (February 20, 2020) and March 31, 2020. We compared hospitalization rates between the study period and two control periods: the corresponding period during the previous year (February 20 to March 31, 2019) and the earlier period during the same year (January 1 to February 19, 2020). Incidence rate ratios comparing the study period with each of the control periods were calculated with the use of Poisson regression. Of the 547 patients who were hospitalized for ACS during the study period, only 127 (23%) were females, accounting for a mean of 3.1 admissions per day, while ACS hospitalized males were 420, with a mean of 10.2 admissions per day. There was a significant decrease driven by a similar reduction in ST-segment elevation myocardial infarction (STEMI) and non-ST-segment elevation myocardial infarction (NSTEMI) diagnosis in both sexes compared to the control periods. A trend toward a greater reduction in admitted females was shown in the intra-year control period (46% admission reduction in females vs 37% in males, with females accounting for 26% of ACS, P=0.10) and a significant reduction when compared to the previous year control period (40% admission reduction in females vs 23% in males, with females accounting for 28% of ACS, P=0.03), mainly related to Unstable Angina diagnosis. The Covid-19 pandemic period closed the gap between men and women in ACS, with similar rates of reduction of hospitalized STEMI and NSTEMI and a trend toward greater reduction in UA admission among women. Furthermore, many typical differences between males and females regarding ischemic heart disease presentations and vessel distribution were leveled.


Inhibition of angiotensin II Gq signaling augments beta-adrenergic receptor mediated effects in a renal artery stenosis model of high blood pressure.
Chronic ventricular pressure overload states, such as hypertension, and elevated levels of neurohormones (norepinephrine, angiotensin II, endothelin-1) initiate cardiac hypertrophy and dysfunction and share the property of being able to bind to Gq-coupled 7-transmembrane receptors. The goal of the current study was to determine the role of endogenous cardiac myocyte Gq signaling and its role in cardiac hypertrophy and dysfunction during high blood pressure (BP). We induced renal artery stenosis for 8 weeks in control mice and mice expressing a peptide inhibitor of Gq signaling (GqI) using a 2 kidney, 1 clip renal artery stenosis model. 8 weeks following chronic high BP, control mice had cardiac hypertrophy and depressed function. Inhibition of cardiomyocyte Gq signaling did not reverse cardiac hypertrophy but attenuated increases in a profile of cardiac profibrotic genes and genes associated with remodeling. Inhibition of Gq signaling also attenuated the loss of cardiac function. We determined that Gq signaling downstream of angiotensin II receptor stimulation negatively impacted beta-adrenergic receptor (AR) responses and inhibition of Gq signaling was sufficient to restore betaAR-mediated responses. Therefore, in this study we found that Gq signaling negatively impacts cardiac function during high BP. Specifically, we found that inhibition of AT1-Gq signaling augmented betaAR mediated effects in a renal artery stenosis model of hypertension. These observations may underlie additional, beneficial effects of angiotensinogen converting enzyme (ACE) inhibitors and angiotensin receptor antagonists observed during times of hemodynamic stress.


Current status of allogeneic bone marrow transplantation.
Use of allogeneic BMTs continues to increase. During the 33-year period between 1955 and 1987, more than 20,000 patients received allogeneic BMTs; more than 50% of these were performed in the 3 years, 1985 through 1987. Transplants are effective therapy for leukemia and other hematologic diseases. They are the treatment of choice for aplastic anemia and chronic myelogenous leukemia, those who fail conventional therapy for acute leukemia, and a variety of immune deficiency disorders. Successful application of BMT is limited by complications such as graft failure, GvHD and interstitial pneumonia, and, until recently, the requirement for an HLA-identical sibling donor. In the past few years, an increasing number of transplants was performed using HLA partially matched related or unrelated donors, with some success. The development of posttransplant complications can often be predicted by risk factor assessment. In this report, current data from the IBMTR are summarized and several risk factors affecting outcome identified.


Interactions between nesfatin-1 and the autonomic nervous system-An overview.
Nesfatin-1, an 82-amino acid polypeptide derived from the precursor protein nucleobindin-2 (NUCB2), was first discovered in 2006 in the rat hypothalamus. The effects and distribution of nesfatin-1 immunopositive neurons in the brain and spinal cord point towards a role of NUCB2/nesfatin-1 in autonomic regulation. Therefore, studies which have been conducted to investigate the interplay between nesfatin-1 and the autonomic nervous system were examined, and the outcomes of this research were summarized. NUCB2/nesfatin-1 immunoreactivity is widely distributed in autonomic centers of the brain and spinal cord in both rodents and humans. In several regions of the hypothalamus, midbrain and brainstem, nesfatin-1 modulates autonomic functions. On the other hand, the autonomic nervous system also influences the activity of nesfatin-1 neurons. Here, the vagus nerve seems to be a crucial factor in the regulation of nesfatin-1. In summary, although data here is still sparse, there is a clear interplay between nesfatin-1 and the autonomic nervous system, the precise clarification of which still requires further research to gain more insight into these complex relationships.


Lirentelimab for severe and chronic forms of allergic conjunctivitis.
Allergic conjunctivitis (AC) is an ocular inflammatory disease with symptoms driven by eosinophils and mast cells. Allergic comorbidities are common. Current treatments are often ineffective in severe AC and limited by potential side effects. Lirentelimab is an anti-sialic acid-binding immunoglobulin-like lectin-8 mAb that depletes eosinophils and inhibits mast cells. We sought to determine safety and preliminary efficacy of lirentelimab in an open-label, phase 1b study. Patients with chronic, severely symptomatic atopic keratoconjunctivitis, vernal keratoconjunctivitis, and perennial AC, and who had history of topical or systemic corticosteroid use, were enrolled to receive up to 6 monthly lirentelimab infusions (dose 1: 0.3 mg/kg, dose 2: 1 mg/kg, subsequent doses: 1 or 3 mg/kg). Changes from baseline in peripheral blood eosinophils, changes in patient-reported symptoms (measured by daily Allergic Conjunctivitis Symptom Questionnaire, including atopic comorbidities), changes in investigator-reported ocular signs and symptoms (Ocular Symptom Scores), changes in quality of life, and changes in tear cytokine and chemokine levels were assessed. Thirty patients were enrolled (atopic keratoconjunctivitis n = 13, vernal keratoconjunctivitis n = 1, perennial AC n = 16), 87% of whom had atopic comorbidities. After lirentelimab treatment, mean improvement was observed in Allergic Conjunctivitis Symptom Questionnaire score (-61%; 95% CI, -75% to -48%) and Ocular Symptom Scores (-53%; 95% CI, -76% to -31%), consistent across atopic keratoconjunctivitis, vernal keratoconjunctivitis, and perennial AC groups. There was substantial improvement in atopic comorbidities, with -55% (95% CI, -78% to -31%), -50% (95% CI, -82% to -19%), and -63% (95% CI, -87% and -38%) reduction in symptoms of atopic dermatitis, asthma, and rhinitis, respectively. Levels of key mediators of inflammation were reduced in patient tears after lirentelimab treatment. The most common adverse effects were mild to moderate infusion-related reactions. Lirentelimab was well tolerated, improved severe AC and concomitant atopic symptoms, and reduced inflammatory mediators in patient tears.


Photodynamic therapy of activated and resting lymphocytes and its antioxidant adaptive response.
In this work we have studied the effects of ALA-mediated photodynamic therapy (PDT) on resting and mitogen-activated murine splenic lymphocytes, evaluating its impact on cell viability. We have also characterised the stress response, measuring the levels of antioxidant enzymes. A 2h exposure to ALA produced 50% lethality upon irradiation of activated cells with 2.1J/cm2. The decrease in cell survival with increasing time exposure to ALA, correlated well with the higher porphyrin accumulation. In resting lymphocytes, in spite of the low amount of porphyrins formed during 2h incubation with ALA, 40% of the cells died after irradiation, this response was not further increased when higher amounts of porphyrins were synthesised. Superoxide dismutase was impaired by light treatment independently of ALA exposure in activated lymphocytes and, to a lesser extent, in resting lymphocytes. PDT induced an antioxidant adaptive response in activated cells 19 h after irradiation, reflected as a net increase in GSHPx activity and a slight reversion of the catalase (CAT) activity already impaired by light treatment. PDT treatment of activated cells also produced a diminution in the GSH/GSSG ratio. Only activated cells are capable of developing an antioxidant adaptive response to PDT treatment.


The health/disease transition process in chronic kidney disease patients: Contributions to nursing care.
To identify the facilitating and hindering conditions of the health/disease transition process in a group of patients with Chronic Kidney Disease undergoing hemodialysis treatment based on the theoretical framework of the Transition Theory. A descriptive study implementing a qualitative approach. Data collection occurred through semi-structured interviews conducted between March and April 2018. The Content Analysis Technique was used for analysis and discussion as a methodological framework and the explanatory model of the Transition Theory as a theoretical framework. Twenty-five (25) patients participated in the study. The conditioning factors of the transition process were identified and organized into three thematic categories: Personal conditioning factors in the transition process; Community conditioning factors in the transition process; and Social conditioning factors in the transition process. A predominance of hindering conditioning factors was evidenced in the personal dimension, while facilitating conditioning factors were shown in relation to the community dimension, and finally the conditioning factors were equal regarding the social dimension.


GABA in regulation of communicative activity and sexual motivation of male mice with different psychoemotional status.
We studied the effects of drugs modulating GABA content in the brain on communicative activity and sexual motivation of male mice. The effects of the drug depended on animal genotype and initial psychoemotional status. Aminooxyacetic acid elevating GABA content did not modulate communicative activity of intact males, reduced it in aggressive animals, restored in anxious animals, and promoted exhaustion of sexual motivation in anxious animals. Thiosemicarbazide reducing GABA level produced an anxiogenic effect and destabilized sexual motivation in intact males.


Epigenetics in lung cancer diagnosis and therapy.
Lung cancer is the leading cause of cancer-related deaths worldwide. The initiation and progression of lung cancer is the result of the interaction between permanent genetic and dynamic epigenetic alterations. DNA methylation is the best studied epigenetic mark in human cancers. Altered DNA methylation in cancer was identified in 1983. Within 30 years of this discovery, DNA methylation inhibitors are used clinically to treat a variety of cancers, highlighting the importance of the epigenetic basis of cancer. In addition, histone modifications, nucleosome remodeling, and micro RNA (miRNA)-mediated gene regulation are also fundamental to tumor genesis. Distinct chromatin alterations occur in all stages of lung cancer, including initiation, growth, and metastasis. Therefore, stage-specific epigenetic changes can be used as powerful and reliable tools for early diagnosis of lung cancer and to monitor patient prognosis. Moreover, since epigenetic changes are dynamic and reversible, chromatin modifiers are promising targets for the development of more effective therapeutic strategies against cancer. This review summarizes the chromatin alterations in lung cancer, focusing on the diagnostic and therapeutic approaches targeting epigenetic modifications that could help to reduce the high case-fatality rate of this dreadful disease.


Lipoma of the corpus callosum: report of a case and review of the literature.
A case of lipoma of the corpus callosum associated with subcutaneous lipoma, whose symptoms were headache and trembling of the extremities, was reported. Roentgenograms showed characteristic findings and computerized tomography revealed a mass of very low density. The literature was reviewed and 15 cases not included in previous compilations were found. Clinical and radiological findings in 89 cases including the present case were analyzed.


Multiple sclerosis in Scandinavia and Finland.
Review of studies of incidence of MS over several decades in seven geographical areas scattered over Scandinavia and Finland have shown that incidence rates fluctuate significantly with time in most of the areas but in an inconsistent pattern. The incidence appeared to the lowest in the northernmost parts of Norway indicating that the well-known north-south gradient for risk of MS does not hold true for high latitudes.


Post-transplant diabetes mellitus and renal cell cancer after renal transplantation.
Diabetes is a risk factor for cancer in the general population. However, few data are available on the association between post-transplant diabetes mellitus (PTDM) and cancer after transplantation. We analyzed this issue in a Spanish cohort of patients without diabetes before transplantation. PTDM was diagnosed with consensus criteria at 12 months after transplantation and 12 months before the diagnosis of cancer. The association between PTDM and cancer (overall and specific types) was evaluated with regression analysis. During a follow-up of 12 years (interquartile range 8-14), 85 cases of 603 developed cancer (829/100 000/year) and 164 (27%) PTDM. The most frequent cancers were renal cell cancer (RCC) n = 15, 146/cases/100 000/year), lung (n = 12, 117/cases/100 000/year), colon (n = 9, 88/cases/100 000/year) and prostate (n = 9, 88/cases/100 000/year). In logistic regression, PTDM was not associated with cancer. Eight of the 164 patients with PTDM (4.9%) vs 7 of the 439 without PTDM developed RCC (1.6%) (P = .027). In multivariate analysis, PTDM was independently associated with RCC [odds ratio (OR) 2.92, confidence interval (CI) 1.03-8.27], adjusting for smoking (OR 4.020, 95% CI 1.34-12.02) and other covariates. PTDM was not associated with other types of cancer. Patients with PTDM must be considered a population at risk for RCC and accordingly, the subject of active surveillance.


Efficacy of retrograde coronary sinus cardioplegia in patients undergoing myocardial revascularization: a prospective randomized trial.
The efficacy of retrograde coronary sinus cardioplegia (RCSC) administered through the right atrium compared with aortic root cardioplegia (ARC) has not been examined critically in patients undergoing coronary artery bypass grafting (CABG). Twenty patients having elective CABG were randomized prospectively to receive cold blood ARC (Group I, 10 patients) or cold blood RCSC (Group II, 10 patients). Patient demographics were similar in both groups. Ventricular function was assessed preoperatively by radionuclide ventriculography and postoperatively by simultaneous hemodynamic and radionuclide ventriculographic studies with volume loading. There was no change in ejection fraction (EF) (preoperative versus postoperative value) in Group I (50 +/- 6% versus 53 +/- 6%) but in group II, at similar peak systolic pressure and similar left ventricular end-diastolic volume index (LVEDVI), LVEF improved significantly (49 +/- 6% versus 60 +/- 12%, p less than 0.05). Postoperative ventricular function (stroke work index versus EDVI) for the left ventricle and right ventricle were similar in both groups. Evaluation of postoperative LV systolic function (end-systolic blood pressure versus end-systolic volume index) and diastolic function (pulmonary capillary wedge pressure versus EDVI) were also similar in both groups. Retrograde coronary sinus cardioplegia is as effective as ARC for intraoperative myocardial protection, and provides excellent postoperative function in patients undergoing elective CABG.


Regional neuromuscular regulation within human rectus femoris muscle during gait in young and elderly men.
Recently, we demonstrated region-specific electromyography (EMG) responses along the rectus femoris (RF) muscle during gait in healthy young men (Watanabe et al., 2014b). For the RF muscle, regional EMG response should be tested to characterize neuromuscular control and/or to assess its dysfunction and/or pathology during gait. We aimed to identify spatial distribution of EMG pattern within the RF muscle in elderly during gait. Seven young men (age: 20.4±1.0 years) and 8 elderly men (age: 73.8±5.9 years) walked on treadmill with three different speed: slow (preferred -1km/h), preferred, and fast (preferred +1km/h). The spatial distribution of surface EMG was tested by central locus activation (CLA), which is calculated from 18 surface electrodes along the longitudinal line of the muscle. CLA were not different between the groups for slow and preferred gait speed (p>0.05) during a gait cycle. In fast gait speed, CLA at 80% of a gait cycle (swing phase) for the elderly were significantly located at more distal site than the young group (p<0.05) (13.0±2.1cm and 10.2±2.2cm from most proximal electrodes for the elderly and young). This difference in CLA reflected a significantly lower EMG activity at the proximal regions in the elderly group (p<0.05). These results suggest the elderly manifest characteristic regional EMG responses within the RF muscle for leg swing movement of fast speed gait.


Serial urinary neutrophil gelatinase associated lipocalin in pediatric diabetic ketoacidosis with acute kidney injury.
Acute kidney injury (AKI) due to Diabetic Ketoacidosis (DKA) is rather common. Novel biomarkers to diagnose AKI are being increasingly used in different settings. The use of urinary Neutrophil Gelatinase-Associated Lipocalin (uNGAL) in predicting persistent AKI in pediatric DKA cases is still not thoroughly investigated. This was a secondary analysis of Saline versus Plasma-Lyte in Ketoacidosis (SPinK) trial data; 66 children (> 1 month-12 years) with DKA, defined by the International Society for Pediatric and Adolescent Diabetes (ISPAD), were analyzed. Children with cerebral edema, chronic kidney disease and those who received pre-referral fluids and/or insulin were excluded. uNGAL and urine NGAL-creatinine ratio (uNCR) at 0 and 24 h were measured in all. Persistent AKI was defined as a composite outcome of continuance of AKI defined by the Kidney Disease Improving Global Outcomes (KDIGO) stage 2 or 3 beyond 48 h from AKI onset, progression of AKI from either KDIGO stage 0 or 1 to a worse stage, need of renal replacement therapy or death. Thirty-five (53%) children had AKI at admission; 32 (91.4%) resolved within 48 h. uNGAL was significantly higher in the AKI group at admission [79.8 ± 27.2 vs 54.6 ± 22.0, p = 0.0002] and at 24 h [61.4 ± 28.3 vs 20.2 ± 14.5, p = 0.0003]. Similar trend was observed with uNCR at admission [6.7 ± 3.7 vs 4.1 ± 2.6, p = 0.002] and at 24 h [6.3 ± 2.5 vs 1.2 ± 1.0, p = 0.01]. Furthermore, uNGAL at admission showed a moderate positive linear correlation with serum creatinine. Additionally, elevated uNGAL at 0 and 24 h correlated with corresponding KDIGO stages. Admission uNGAL >88 ng/ml and uNCR of >11.3 ng/mg had a sensitivity of 66% and 67%, specificity of 76% and 95%, and Area under the receiver operating characteristic curve (AUC) of 0.78 and 0.89 respectively for predicting persistent AKI at 48 h. Majority of AKI resolved with fluid therapy. While uNGAL and uNCR both correlated with serum creatinine and AKI stages, serial uNCR was a better predictor of persistent AKI than uNGAL alone. However, feasibility of routine uNGAL measurement to predict persistent AKI in DKA needs further elucidation. This was a secondary analysis of the data of SPinK trial [CTRI/2018/05/014042 ( ctri.nic.in )].


Application of taste reactivity to study the mechanism of alcohol intake inhibition by the tachykinin aminosenktide.
Tachykinin NK-3 receptor agonists reduce alcohol intake in alcohol-preferring rats; the nucleus basalis magnocellularis (NBM) is highly sensitive to their effect. Tachykinins and their receptors are widely distributed in gustatory pathways and NK-3 receptor agonists have been reported to modify taste reactivity to salt solutions in rats. The present study evaluated whether the TK NK-3 receptor agonist aminosenktide (NH2-SENK) influences taste reactivity to ethanol solutions. Genetically selected Marchigian Sardinian alcohol-preferring (msP) rats were employed. In response to the intraoral infusion (0.8 ml in 1 min) of 10, 20, 40, or even 60% ethanol solution, ethanol-naive rats showed a large number of ingestive reactions and a much lower number of aversive reactions. Two min before the intraoral infusion of 10 or 40% ethanol, NH2-SENK was injected either into the lateral ventricle (LV) or into the NBM. Doses of NH2-SENK that markedly reduce alcohol intake, 125 ng/rat into the LV or 5 ng/site into the NBM, did not modify the ingestive reactions and, in some instances, reduced the aversive reactions to ethanol solutions in ethanol-naive rats. Injections of 125 ng/rat into the LV failed to modify taste reactions in ethanol-experienced rats. The present results show that msP rats have an innate hedonic evaluation of ethanol solutions, even of high concentration. Moreover, they indicate that reduction of ethanol intake induced by TK NK-3 receptor agonists in alcohol-preferring rats does not depend on influences on gustatory processes.


Deciphering the conformational transitions of LIMK2 active and inactive states to ponder specific druggable states through microsecond scale molecular dynamics simulation.
LIMK2 inhibitors are one of the potential therapeutic modalities for treating various diseases. In the current scenario, there is a paucity of effective LIMK inhibitors that are highly specific with minimal off-target effects. To date, the conformational transitions of LIMK2 from DFGinαCin (CIDI) (active) to DFGoutαCout (CODO) (inactive) states are yet to be probed and are essential for capturing the unique, druggable conformations. Therefore, this study was intended to capture the diverse conformational states of LIMK2 for accelerating the rational identification of conformation specific inhibitors through high-end structural bioinformatics protocols. Hence, in this study, molecular modelling followed by an extensive microsecond timescale of molecular dynamics simulation was performed encompassing perturbation response scanning, metapath, and community analysis towards the conformational sampling of LIMK2. Overall this study precisely identifies the conformational ensemble of LIMK2 the intermediate inactive states namely, CIDO, CinterDinter, CIDinter, CinterDI, CinterDO, CODI, CODinter apart from CIDI and CODO. This also facilitated observing that β8 preceding XDFG, αC (F373, L374), and αD (L413) as the major effectors that may facilitate the regulation of varying conformational transitions among the states. Additionally, the conserved β sheets and the loops namely, C.l, b.l, and G/P.loop were observed to be involved in the metapath for allosteric communication among the intermediates with CIDI and CODO state. Moreover, only the CODO state was observed to have closed type A.l, while the CIDI and other intermediate states except for CIDO were observed to have open-DFG out type A.l, thereby enabling the binding of substrate. Apart from these, the druggable site analysis inferred that the CIDI and CODO states harbor prominent druggable sites spanning the conserved N-lobe, while the intermediates were observed to have unraveled allosteric druggable sites distal from the ATP binding site, majorly spanning the C-lobe of LIMK2. Thus, this study provides potential insights into the intermediate conformational druggable states of LIMK2 and also the druggable conformations, especially the inactive states of LIMK2, as a specific therapeutic targeting mode. Thus, providing a widened avenue to ponder the allosteric sites or the isoform selectivity conformations for targeting LIMK2 in various disease conditions.


Development and Preclinical Investigation of Physically Cross-Linked and pH-Sensitive Polymeric Gels as Potential Vaginal Contraceptives.
This study explored the development of cross-linked gels to potentially provide a physical barrier to vaginal sperm transport for contraception. Two types of gels were formulated, a physically cross-linked iota-carrageenan (Ci) phenylboronic acid functionalized hydroxylpropylmethyacrylate copolymer (PBA)-based (Ci-PBA) gel, designed to block vaginal sperm transport. The second gel was pH-shifting cross-linked Ci-polyvinyl alcohol-boric acid (Ci-PVA-BA) gel, designed to modulate its properties in forming a viscoelastic, weakly cross-linked transient network (due to Ci gelling properties) on vaginal application (at acidic pH of ~3.5-4.5) to a more elastic, densely cross-linked (due to borate-diol cross-linking) gel network at basic pH of 7-8 of seminal fluid, thereby acting as a physical barrier to motile sperm. The gels were characterized for dynamic rheology, physicochemical properties, and impact on sperm functionality (motility, viability, penetration). The rheology data confirmed that the Ci-PBA gel was formed by ionic interactions whereas Ci-PVA-BA gel was chemically cross-linked and became more elastic at basic pH. Based on the screening data, lead gels were selected for in vitro sperm functionality testing. The in vitro results confirmed that the Ci-PBA and Ci-PVA-BA gels created a barrier at the sperm-gel interface, providing sperm blocking properties. For preclinical proof-of-concept, the Ci-PBA gels were applied vaginally and tested for contraceptive efficacy in rabbits, demonstrating only partial efficacy (40-60%). Overall, the in vitro and in vivo results support the development and further optimization of cross-linked gels using commercially available materials as vaginal contraceptives.


Afferent modulation of dopamine neuron firing differentially regulates tonic and phasic dopamine transmission.
The mesolimbic dopamine system is centrally involved in reward and goal-directed behavior, and it has been implicated in multiple psychiatric disorders. Understanding the mechanism by which dopamine participates in these activities requires comprehension of the dynamics of dopamine release. Here we report dissociable regulation of dopamine neuron discharge by two separate afferent systems in rats; inhibition of pallidal afferents selectively increased the population activity of dopamine neurons, whereas activation of pedunculopontine inputs increased burst firing. Only the increase in population activity increased ventral striatal dopamine efflux. After blockade of dopamine reuptake, however, enhanced bursting increased dopamine efflux three times more than did enhanced population activity. These results provide insight into multiple regulatory systems that modulate dopamine system function: burst firing induces massive synaptic dopamine release, which is rapidly removed by reuptake before escaping the synaptic cleft, whereas increased population activity modulates tonic extrasynaptic dopamine levels that are less influenced by reuptake.