Systemic therapy for advanced clear cell renal cell carcinoma after discontinuation of immune-oncology and VEGF targeted therapy combinations.
Several phase 3 studies reported positive results for combinations of Immune-Oncology (IO) and Vascular Endothelial Growth Factor (VEGF) targeted therapies in patients with metastatic clear cell Renal Cell Carcinoma (ccRCC). However, there are limited data on outcomes to systemic therapy after IO-VEGF combinations. A retrospective analysis was performed on patients with metastatic ccRCC treated at the Memorial Sloan Kettering Cancer Center and Cleveland Clinic who initiated systemic therapy post IO-VEGF including combinations with VEGF receptor (VEGFR) tyrosine kinase inhibitors (IO-TKI) and combinations with the anti-VEGF monoclonal antibody bevacizumab (IO-Bev). The study objectives were to evaluate the objective response rate (ORR), progression-free survival (PFS) and overall survival (OS) on systemic therapy post IO-VEGF. RECIST v1.1 criteria were used to determine radiological responses and progression. Survival estimates were evaluated with the Kaplan-Meier methods and the log-rank test from the start of systemic therapy post IO-VEGF to the event of interest. A total of fifty-nine patients were treated post discontinuation of IO-VEGF regimens which included IO-Bev (n = 35; 59%) and IO-TKI (n = 24; 41%). Fifty-eight patients (98%) received IO-VEGF regimens as part of a clinical trial. Subsequent therapies included cabozantinib (n = 22; 37%), axitinib (n = 18; 31%), pazopanib (n = 4; 7%), lenvatinib and everolimus (n = 4; 7%), mTOR inhibitor monotherapy (n = 3; 5%), axitinib and dalantercept (n = 2; 3%), sunitinib (n = 1; 2%), sorafenib (n = 1; 2%), and treatment with agents on unreported clinical trials (n = 4; 7%). Patients treated on unreported clinical trials were excluded from the efficacy analysis. Post IO-VEGF, the ORR was 25% and median PFS was 12.0 months (95% CI, 8.2-24.5). Median OS was 24.5 months (95% CI, 12-NE) and 12 months OS rate was 63.3% (95% CI, 48.6-74.9). We observed no differences post IO-VEGF OS when comparing IO- TKI vs IO-Bev (Log-rank p = 0.73). Post IO-VEGF, most patients received VEGFR-TKIs. In this setting, VEGFR-TKIs demonstrated clinical activity and remain a viable option for salvage therapy after progression on IO-VEGF.


Nicotine-induced monoamine neurotransmitter changes in the brain of young rats.
A number of studies in various species including man indicated a greater risk of drug preference and addictive behavior in young as compared to adults. Such age dependent preference was also found with nicotine. To examine possible mechanisms for this difference in our continuing study of reward mechanisms, we compared nicotine-induced neurotransmitter changes in the brain regions of adult and young Sprague-Dawley rats, assaying the transmitters via microdialysis in conscious freely moving animals. In general, nicotine-induced changes were significantly less in the regions measured in the young. Nicotine-induced effects on dopamine in the dorsal and ventral hippocampus (VH), prefrontal and medial temporal cortex, and superior cerebral peduncle were lower in the young than in adult, the same in the ventral tegmental area (VTA) and lateral septal nucleus (LS), and somewhat higher in the nucleus accumbens shell (NAccS). Norepinephrine levels in the young were lower in all areas except in the VH where they were the same, and serotonin levels were lower except in the VTA and LS where they remained the same, and higher in the NAccS. Age-dependent differences in the metabolites measured were more mixed. We conclude that the greater nicotine preference in young is not paralleled by a greater effect of nicotine on the release of monoamines at least in most of the brain areas assayed. Thus, increases of nicotine reward are not likely due to increases of monoamines in reward and cognitive areas. The small increase of dopamine (DA) and more significant increase of serotonin (5-HT) only in the NAccS are of significance, and would indicate a more significant role of 5-HT than of DA at least in the age difference in nicotine preference. Developmental changes in receptor composition and distribution involving several transmitter systems and other components such as neuropeptides are also likely to play a role.


Magnetic bead-based separation of pneumococcal serotypes.
The separation of pneumococcal serotypes from a complex polymicrobial mixture may be required for different applications. For instance, a minority strain could be present at a low frequency in a clinical sample, making it difficult to identify and isolate by traditional culture-based methods. We therefore developed an assay to separate mixed pneumococcal samples using serotype-specific antiserum and a magnetic bead-based separation method. Using qPCR and colony counting methods, we first show that serotypes (12F, 23F, 3, 14, 19A, and 15A) present at ∼0.1% of a dual serotype mixture can be enriched to between 10% and 90% of the final sample. We demonstrate two applications for this method: extraction of known pneumococcal serotypes from saliva samples and efficient purification of capsule switch variants from experimental transformation experiments. This method may have further laboratory or clinical applications when the selection of specific serotypes is required.


Life-cycle energy efficiency and environmental impacts of bioethanol production from sweet potato.
Life-cycle assessment (LCA) was used to evaluate the energy efficiency and environmental impacts of sweet potato-based bioethanol production. The scope covered all stages in the life cycle of bioethanol production, including the cultivation and treatment, transport, as well as bioethanol conversion of sweet potato. Results show that the net energy ratio of sweet potato-based bioethanol is 1.48 and the net energy gain is 6.55 MJ/L. Eutrophication is identified as the most significant environmental impact category, followed by acidification, global warming, human toxicity, and photochemical oxidation. Sensitivity analysis reveals that steam consumption during bioethanol conversion exerts the most effect on the results, followed by sweet potato yields and fertilizers input. It is suggested that substituting coal with cleaner energy for steam generation in bioethanol conversion stage and promotion of better management practices in sweet potato cultivation stage could lead to a significant improvement of energy and environmental performance.


New granulocyte antigens demonstrated by microgranulocytotoxicity assay.
Although granulocyte transfusions and bone marrow transplantation are becoming common clinical modalities, our knowledge of surface nonerythroid, nonlymphoid, non-HLA hematopoetic antigens remains very incomplete. Accordingly, we have systematically screened sera from recipients of multiple granulocyte and whole blood transfusions, and immunoneutropenic patients for antibodies directed primarily at granulocytes. The initial screens demonstrated that >50% of the sera from the above sources contained non-HLA cytotoxic and/or agglutinating antibodies. Preliminary clustering indicated seven possible new specificities detected by microgranulocytotoxicity. Calculations for Hardy-Weinberg goodness of fit based on a study of 98 unrelated donors plus informative families established that 5 of these were alleles of a single new locus termed Human Granulocyte Antigen (HGA)-3a, b, c, d, and e. Absorptions indicated that these antigens were present on mature granulocytes but absent from platelets, lymphocytes, monocytes, and myeloid precursors. A single antigen of another separate locus, HGA-1, was also identified. Absorptions revealed a quite different distribution for HGA-1 than HGA-3, this antigen being detected on monocytes and myeloblasts as well as on mature granulocytes. Independent segregation of the three loci from HLA, from the NA-NB and the 5a-5b antigens, and from themselves was confirmed in informative families.Finally, it seems likely that other antigens will be identified because several other sera that react with both monocytes and granulocytes have been detected.


Ruminal In Vitro Protein Degradation and Apparent Digestibility of Energy and Nutrients in Sheep Fed Native or Ensiled + Toasted Pea (Pisum sativum) Grains.
Pea grains may partially replace soybean or rapeseed meals and cereals in ruminant diets, but substitution by unprocessed peas is limited by high ruminal protein solubility. The effect of combined ensiling and toasting of peas using a mobile toaster (100 kg/h throughput rate, 180 to 190 °C supplied air temperature) on rumen-undegraded protein (RUP) was tested in vitro using the Streptomyces griseus protease test. The effects of ensiling plus toasting on apparent digestibility of organic matter (OM), gross energy (GE), and proximate nutrients were examined in a digestion trial. Concentrations of metabolizable energy (ME) and net energy lactation (NEL) were calculated. Native peas had 38 g RUP/kg dry matter (DM), which was 20% of crude protein (CP). Rumen-undegraded protein increased three-fold after ensiling plus toasting (p < 0.001). Acid detergent insoluble protein increased five-fold. Apparent digestibility was 0.94 (OM), 0.90 (CP), and above 0.99 (nitrogen-free extract, starch, and sugars) and was not altered by the treatment. The ME (13.9 MJ/kg DM) or the NEL (8.9 MJ/kg DM) concentration was similar in native and ensiled plus toasted peas. This technique can easily be applied on farms and may increase RUP. However, it needs to be clarified under which conditions pea protein will be damaged.


A cephalometric study of velar stretch in adolescent subjects.
Cephalometric x-rays were used to evaluate velar stretch in 20 normal adolescents during the production of /s/. Radiopaque markers were used so that the behaviors of both the anterior and posterior portions and the total velum cound be evaluated. Velar stretch was found in each of the subjects with the average stretch (15.19%) greater than the amount reported for 10-year-old subjects but less than the average stretch for adults. The correlation between amount of velar stretch and need ratio was not significant. The different stages of involution of adenoid tissue, which is characteristic of this age group, resulted in varying configurations of the naso-pharynx were postulated as one possible explanation for this unexpected finding. Normative data and clinical implications are presented.


Rapid normalization of marked dilatation of the cerebral duro-venous system in a newborn infant mimicking a great vein of Galen varix.
A newborn infant with a marked dilatation of the cerebral duro-venous system is presented. The patient was diagnosed as having a vein of Galen aneurysmal varix by a cranial ultrasound examination immediately following delivery. Computed tomographic angiography on the following day, however, showed a marked dilatation of the cerebral duro-venous system, including the great vein of Galen, superior sagittal sinus, torcular herophili and transverse sinuses. There were no arteriovenous fistulas at the vein of Galen. Dilatation of the duro-venous system and concomitant heart failure subsided rapidly after intravenous administration of indomethacin for the treatment of the patent ductus arteriosus on the fourth day of life. Dilatation of the duro-venous system in a newborn infant should be differentiated from any form of vein of Galen aneurysm.


Epidural pressures and spread of 2% lidocaine in the epidural space: influence of volume and speed of injection of the local anesthetic solution.
The impact of epidural pressures on the spread of epidural block is controversial. This study examined the effect of volume and speed of injection of local anesthetics on epidural pressures and the spread of anesthesia. Twenty women undergoing epidural anesthesia received a test dose of 3 mL of 2% lidocaine with epinephrine 1:200,000 over 6 seconds and subsequently the main dose of 15 mL of the solution over 30 seconds (group I) or 3 minutes (group II). Variables studied included epidural pressures and the extent and duration of thermal and sensory block. Peak epidural pressure following the main dose was significantly higher in group I as compared to group II. All other epidural pressures and the extent and duration of sensory and thermal block were similar in both groups. The peak epidural pressure in group II and the pressures obtained at 3 minutes after the main dose in both groups were inversely correlated with the extent of the thermal block and directly correlated with its regression time. The peak epidural pressures correlated with the speed of injection of the lidocaine solution and not with its volume, whereas the remaining epidural pressures correlated with its volume and not with the speed of injection. The extent and duration of the thermal block exhibited a more consistent correlation (inverse and direct, respectively) with the epidural pressures than those of the sensory block.


On-line monitoring of cell mass in mammalian cell cultures by acoustic densitometry.
Acoustic resonance densitometry (ARD) provides a highly reproducible and stable method for on-line measurement of culture biomass density. The technique provides a direct determination of changes in relative density of culture medium and cell mass. At cell concentrations higher than 10(6) cells mL(-1)this method can replace cell counts and provide a continuous measure of total cell mass. In cultures of hybridomas or U937 human lymphoma cells, the ARD value correlates well with cell number except when the average cell size changes during culture. It is argued that cell mass determined by ARD rather than cell number should be used as the basis for measurements of specific biological activity.


Random replication of the stringent plasmid R1 in Escherichia coli K-12.
The R-factor R1 is present in a low number of copies per genome (near unity, so-called stringent control of replication). The replication of R1 was studied in a density-shift experiment. One generation after the shift about 20% of the R1 copies had not replicated, whereas about 20% had replicated at least twice. The results are in quantitative accordance with a random replication of R1 in which the replicating molecules are taken from a cytoplasmic plasmid pool and transferred back to the pool after replication. This is analogous to the results obtained by Bazaral and Helinski (1970) and Rownd (1969) for plasmids that are present in 10 to 20 copies per genome (so-called relaxed control of replication). Hence, there seem to be no difference between stringent and relaxed plasmids with respect to selection of plasmid molecules for replication. However, we cannot tell whether all R1 copies in a cell replicate during a fraction of or throughout the cell cycle. The random selction of plasmid copies for replication has to be considered when models for control of replication are constructed.


Different transcriptional control of metabolism and extracellular matrix in visceral and subcutaneous fat of obese and rimonabant treated mice.
The visceral (VAT) and subcutaneous (SCAT) adipose tissues play different roles in physiology and obesity. The molecular mechanisms underlying their expansion in obesity and following body weight reduction are poorly defined. C57Bl/6 mice fed a high fat diet (HFD) for 6 months developed low, medium, or high body weight as compared to normal chow fed mice. Mice from each groups were then treated with the cannabinoid receptor 1 antagonist rimonabant or vehicle for 24 days to normalize their body weight. Transcriptomic data for visceral and subcutaneous adipose tissues from each group of mice were obtained and analyzed to identify: i) genes regulated by HFD irrespective of body weight, ii) genes whose expression correlated with body weight, iii) the biological processes activated in each tissue using gene set enrichment analysis (GSEA), iv) the transcriptional programs affected by rimonabant. In VAT, "metabolic" genes encoding enzymes for lipid and steroid biosynthesis and glucose catabolism were down-regulated irrespective of body weight whereas "structure" genes controlling cell architecture and tissue remodeling had expression levels correlated with body weight. In SCAT, the identified "metabolic" and "structure" genes were mostly different from those identified in VAT and were regulated irrespective of body weight. GSEA indicated active adipogenesis in both tissues but a more prominent involvement of tissue stroma in VAT than in SCAT. Rimonabant treatment normalized most gene expression but further reduced oxidative phosphorylation gene expression in SCAT but not in VAT. VAT and SCAT show strikingly different gene expression programs in response to high fat diet and rimonabant treatment. Our results may lead to identification of therapeutic targets acting on specific fat depots to control obesity.


Functionality of system components: conservation of protein function in protein feature space.
Many protein features useful for prediction of protein function can be predicted from sequence, including posttranslational modifications, subcellular localization, and physical/chemical properties. We show here that such protein features are more conserved among orthologs than paralogs, indicating they are crucial for protein function and thus subject to selective pressure. This means that a function prediction method based on sequence-derived features may be able to discriminate between proteins with different function even when they have highly similar structure. Also, such a method is likely to perform well on organisms other than the one on which it was trained. We evaluate the performance of such a method, ProtFun, which relies on protein features as its sole input, and show that the method gives similar performance for most eukaryotes and performs much better than anticipated on archaea and bacteria. From this analysis, we conclude that for the posttranslational modifications studied, both the cellular use and the sequence motifs are conserved within Eukarya.


Executive functions in the evaluation of accident risk of older drivers.
The main objective of these studies was to analyse the difference in driving attitude and aptitude, between two groups of elderly male drivers (65 years or more), one being accident-free and the second having three accidents or more in the last 5 years. The first study compared the driving habits of 90 older accident-free drivers with 90 drivers having a history of accidents. The second study, on a subgroup of 60 of the original 180 subjects (30 accident-free and 30 having accidents), compared cognitive function, with particular emphasis on executive functions as measured by neuropsychological tests, and attitude and self-reported driving behaviour. The results show that elderly drivers having a history of accidents, compared to the control group: (1) have poorer performance on the four cognitive measurements of executive functions; (2) report to have more prudent behaviour on the road (e.g., reducing their speed); and (3) have the intention to adopt less risky driving behaviour. This study suggests that a subgroup of the older driver population has cognitive problems and driving disabilities that cannot be compensated by apparently more careful behaviour on the road. The results confirm the importance of proper assessment of cognitive processes and underscore the potential of measuring executive functions for the evaluation of driving competence of elderly persons.


Recent advances in pharmacovigilance of antiretroviral therapy in HIV-infected and exposed children.
Antiretroviral therapy (ART) has greatly improved the survival of HIV-infected children. However, ART is associated with immediate and long-term adverse events. Pharmacovigilance systems, although imperfect, have been developed in many high-income countries (HICs), but coverage in low- and middle-income countries (LMICs) is poor and uneven. This review covers the recent advances in the understanding of adverse events following perinatal ART exposure, including surveillance from birth cohorts; we also describe the adverse events of antiretroviral drugs among HIV-infected children, focussing particularly on those relevant to LMICs, where more than 90% of HIV-infected children live. ART is largely safe in both HIV-infected and HIV-exposed uninfected children, in whom no significant increase in birth defects has been noted. Among HIV-infected children, toxicity to some drugs may be less frequent than in adults, possibly related to immature immune systems in younger children. As per WHO guidelines, many countries are moving from stavudine-based to zidovudine-based or abacavir-based fixed-dose combination (with nevirapine/lamivudine) paediatric mini-pills. However, reassuring data are emerging about short-term stavudine use in LMICs, as this remains an important first-line regimen for young children, as well as an alternative to zidovudine for anaemic children. Zidovudine appears to be well tolerated in young children living in nonmalarious areas, and, among African children, concerns about abacavir hypersensitivity have not been substantiated. Optimization of first-line ART regimens needs to take account of the toxicities in HIV-infected children, in particular as they will take ART much longer than adults and during the period of growth and development. The benefits of ART in pregnancy are clear, but long-term follow-up of ART-exposed infants in LMICs through integrated surveillance systems would be invaluable.


Bone marrow transplantation for chronic myelogenous leukemia.
Chronic myelogenous leukemia (CML) is a hematologic malignancy characterized by an initial chronic phase of expanded clonal myelopoiesis followed by inevitable progression to accelerated phase and finally to fatal blast crisis. Although interferon therapy results in hematologic control of disease in most patients and major cytogenetic responses in 30% to 35%, resulting in better survival than with conventional therapy with hydroxyurea or busulfan, most patients have residual disease detectable by molecular methods. Allogeneic bone marrow transplantation may cure selected patients with CML and should be considered early in the course of disease. Newer approaches to therapy include autografting using mobilized peripheral blood stem cells. Selection and expansion of nonleukemic progenitors may improve outcome following autologous transplantation. Advances in understanding of mechanisms of action of antisense oligonucleotides, ribozymes, and tyrosine kinase inhibitors and the immunologic responses to CML cells are also likely to lead to novel approaches to therapy.


Overestimation of canine albumin concentration with the bromcresol green method in heparinized plasma samples.
Albumin concentrations are routinely measured in dogs with bromcresol green (BCG)-binding assays on automated chemistry analyzers. Several variables affect this assay, including the length of reaction time, sample type, and lack of specificity of BCG for albumin. We observed that albumin concentrations measured with BCG appeared higher in heparinized plasma samples in sick dogs. The objective of this study was to determine the effect of anticoagulant and assay procedure on BCG albumin concentrations in clinically ill dogs. We hypothesized that albumin concentrations would be overestimated in heparinized plasma compared with serum because of the combination of heparin and fibrinogen. Furthermore, we hypothesized that the overestimation would be influenced by assay parameters. Blood was collected from 32 clinically ill dogs into tubes containing heparin, citrate, or no anticoagulant. Citrate was chosen to assess the effect of fibrinogen in the absence of heparin. Albumin concentration was measured in all 3 sample types from each dog using 2 different BCG procedures on an automated chemistry analyzer. The BCG procedures (standard and modified) differed in the wavelengths used for absorbance readings (standard, 600/700; modified, 570/505) and the time point at which absorbance was measured (standard, 100 seconds; modified, 40 seconds). In addition, the modified method incorporated a sample blank. Globulin fractions, fibrinogen concentration, and indices of lipemia, hemolysis, and icterus were evaluated for their contribution to the overestimation of albumin concentration in heparinized plasma compared with serum samples. Albumin concentrations were significantly higher (P </=.05) in heparinized plasma (mean +/- SE, 3.8 +/- 0.1 g/dL) than in serum (3.6 +/- 0.2 g/dL) or citrated plasma (3.2 +/- 0.1 g/dL). Overestimation was evident only with the standard BCG procedure. Multiple linear regression analysis indicated that fibrinogen was largely responsible for the higher albumin concentration in heparinized plasma with the standard method. Based on these results, heparin is not recommended as an anticoagulant for albumin measurement in dogs when using a BCG method unless the assay uses a sample blank and a reaction time of <1 minute.


Development of Phaleria macrocarpa (Scheff.) Boerl Fruits Using Response Surface Methodology Focused on Phenolics, Flavonoids and Antioxidant Properties.
In this study, the optimal conditions for the extraction of antioxidants from the Buah Mahkota Dewa fruit (Phaleria macrocarpa) was determined by using Response Surface Methodology (RSM). The optimisation was applied using a Central Composite Design (CCD) to investigate the effect of three independent variables, namely extraction temperature (°C), extraction time (minutes) and extraction solvent to-feed ratio (% v/v) on four responses: free radical scavenging activity (DPPH), ferric ion reducing power assay (FRAP), total phenolic content (TPC) and total flavonoid content (TFC). The optimal conditions for the antioxidants extraction were found to be 64 °C extraction temperature, 66 min extraction time and 75% v/v solvent to-feed ratio giving the highest percentage yields of DPPH, FRAP, TPC and TFC of 86.85%, 7.47%, 292.86 mg/g and 3.22 mg/g, respectively. Moreover, the data were subjected to Response Surface Methodology (RSM) and the results showed that the polynomial equations for all models were significant, did not show lack of fit, and presented adjusted determination coefficients (R²) above 99%, proving that the yield of phenolic, flavonoid and antioxidants activities obtained experimentally were close to the predicted values and the suitability of the model employed in RSM to optimise the extraction conditions. Hence, in this study, the fruit from P. macrocarpa could be considered to have strong antioxidant ability and can be used in various cosmeceutical or medicinal applications.


Transient receptor potential TRPM3 channels: Pharmacology, signaling, and biological functions.
The transient receptor potential melastatin-3 (TRPM3) channel belongs to the family of transient receptor potential (TRP) cation channels that are expressed in a variety of tissues and cell types, including dorsal root ganglia, cardiomyocytes and pancreatic beta-cells. Although its natural ligands are currently unknown, TRPM3 channels can be activated by the neurosteroid pregnenolone sulfate, the synthetic ligand CIM0216, and by noxious heat. TRPM3 channels are regulated by phosphoinositides, and perhaps by calmodulin. Stimulation of TRPM3 induces an intracellular signaling cascade involving a rise in intracellular Ca2+, activation of the protein kinases Raf, ERK and JNK, and the activation of the stimulus-responsive transcription factors AP-1, CREB, Egr-1, and Elk-1. Functionally, stimulation of TRPM3 channels is connected with heat sensation by somatosensory neurons, insulin secretion by pancreatic beta-cells, regulation of neurotransmitter release, iris constriction, and tumor promotion. With the development of highly specific activators and inhibitors of TRPM3 channels, we expect that additional tissue-specific functions of TRPM3 channels will be discovered, establishing TRPM3 channels as a new therapeutic target.


The microbiome of the lung.
Investigation of the lung microbiome is a relatively new field. Although the lungs were classically believed to be sterile, recently published investigations have identified microbial communities in the lungs of healthy humans. At the present time, there are significant methodologic and technical hurdles that must be addressed in ongoing investigations, including distinguishing the microbiota of the upper and lower respiratory tracts. However, characterization of the lung microbiome is likely to provide important pathogenic insights into cystic fibrosis, respiratory disease of the newborn, chronic obstructive pulmonary disease, and asthma. In addition to characterization of the lung microbiome, the microbiota of the gastrointestinal tract have profound influence on the development and maintenance of lung immunity and inflammation. Further study of gastrointestinal-respiratory interactions is likely to yield important insights into the pathogenesis of pulmonary diseases, including asthma. As this field advances over the next several years, we anticipate that studies using larger cohorts, multicenter designs, and longitudinal sampling will add to our knowledge and understanding of the lung microbiome.


Increase of serotype 8, ST53 clone, as the prevalent strain of Streptococcus pneumoniae causing invasive disease in Madrid, Spain (2012-2015).
In recent years, Streptococcus pneumoniae serotype 8 has become the most prevalent cause of invasive pneumococcal disease (IPD) in Madrid, Spain. The objective of this study was to characterize the invasive clones of S. pneumoniae serotype 8 in Madrid over the 2012-2015 period. From January 2012 to December 2015, a total of 1543 invasive isolates were studied. Serotyping was carried out by Pneumotest-Latex agglutination and Quellung reaction. Susceptibilities to penicillin, erythromycin and levofloxacin were determined by the Etest®. All serotype 8 strains were typed by multilocus sequence typing (MLST) and by pulsed-field gel electrophoresis (PFGE). Two hundred and forty-eight (248) serotype 8 strains were detected (16.1%) and 243 of them were available for molecular typing. Nine sequence types (STs) by MLST (8-ST53, 8-ST63, 8-ST404, 8-ST1107, 8-ST989, 8-ST1110, 8-ST2231, 8-ST3544 and 8-ST4301), and nine PFGE profiles were identified (one corresponding to each ST). The 8-ST53 clone was the most widespread, and increased from 53.8% among all serotype 8 isolates in 2012, to 90.1% in 2015. In contrast, the 8-ST63 clone, resistant to levofloxacin and erythromycin, decreased from 30.8%, among all serotype 8 strains in 2012, to 5.0% in 2015. The increase in our region of S. pneumoniae serotype 8, not included in conjugated vaccines, occurred at the expense of the 8-ST53 clone. On the contrary, the 8-ST63 clone decreased. Since clone 8-ST63 has the theoretical advantage of its levofloxacin-erythromycin resistance in comparison to 8-ST53, the predominance of 8-ST53 over 8-ST63 is striking.


Conventional alpha beta (αβ) T cells do not contribute to acute intestinal ischemia-reperfusion injury in mice.
Ischemia-reperfusion injury (IRI) is associated with significant patient mortality and morbidity. The complex cascade of IRI is incompletely understood, but inflammation is known to be a key mediator. In addition to the predominant innate immune responses, previous research has also indicated that αβ T cells contribute to IRI in various organ models. The aim of this study was to clarify the role αβ T cells play in IRI to the gut. Adult wild-type (WT) and αβ T cell-deficient mice were subjected to acute intestinal IRI with 30min ischemia followed by 4h reperfusion. The gene expression of pro-inflammatory cytokines was measured by qPCR, and the influx of leukocyte subpopulations in the gut was assessed via flow cytometry and histology. Pro-inflammatory cytokines in the serum were measured, and transaminases were assessed as an indicator of distant organ IRI. Intestinal IRI led to an increased expression of pro-inflammatory cytokines in the gut tissue and an influx of leukocytes that predominantly consisted of neutrophils and macrophages. Furthermore, intestinal IRI increased serum IL-6, TNF-α, and ALT/AST levels. The αβ T cell-deficient mice did not exhibit a more significant increase in pro-inflammatory cytokines in the gut or serum following IR than the WT mice. There was also no difference between WT- and αβ T cell-deficient mice in terms of neutrophil infiltration or macrophage activation. Furthermore, the increase in transaminases was equal in both groups indicating that the level of distant organ injury was comparable. An increasing body of evidence demonstrates that αβ T cells play a key role in IRI. In the gut, however, αβ T cells are not pivotal in the first hours following acute IRI as deficiency does not impact cytokine production, neutrophil recruitment, macrophage activation, or distant organ injury. Thus, αβ T cells may be considered innocent bystanders during the acute phase of intestinal IRI.


Li-Doped Bioactive Ceramics: Promising Biomaterials for Tissue Engineering and Regenerative Medicine.
Lithium (Li) is a metal with critical therapeutic properties ranging from the treatment of bipolar depression to antibacterial, anticancer, antiviral and pro-regenerative effects. This element can be incorporated into the structure of various biomaterials through the inclusion of Li chloride/carbonate into polymeric matrices or being doped in bioceramics. The biocompatibility and multifunctionality of Li-doped bioceramics present many opportunities for biomedical researchers and clinicians. Li-doped bioceramics (capable of immunomodulation) have been used extensively for bone and tooth regeneration, and they have great potential for cartilage/nerve regeneration, osteochondral repair, and wound healing. The synergistic effect of Li in combination with other anticancer drugs as well as the anticancer properties of Li underline the rationale that bioceramics doped with Li may be impactful in cancer treatments. The role of Li in autophagy may explain its impact in regenerative, antiviral, and anticancer research. The combination of Li-doped bioceramics with polymers can provide new biomaterials with suitable flexibility, especially as bio-ink used in 3D printing for clinical applications of tissue engineering. Such Li-doped biomaterials have significant clinical potential in the foreseeable future.


Fatal Intracranial Hemorrhage Associated with Oral Warfarin Use.
Intracerebral hemorrhage is the most devastating complication in patients taking oral warfarin. Despite theoretic reversibility with fresh frozen plasma, vitamin K and prothrombin complex concentrate, it remains an entity with high mortality. Fluid-fluid level, also known as blood-fluid level, sedimentation level, or hematocrit effect, seen on noncontrast computed tomography (CT) scan is a characteristic finding associated with patients who have coagulopathy or who are receiving oral anticoagulation therapy. We present the case of an 80-year-old female patient requiring long-term anticoagulation due to atrial fibrillation, who presented acute neurological symptoms including thunderclap headache and hemiparesis. Urgent noncontrast CT scan showed classic signs of warfarin-induced intracranial hemorrhage with a fatal outcome a few hours after admission.


Measuring physician practice, preparedness and preferences for genomic medicine: a national survey.
Even as genomic medicine is implemented globally, there remains a lack of rigorous, national assessments of physicians' current genomic practice and continuing genomics education needs. The aim of this study was to address this gap. A cross-sectional survey, informed by qualitative data and behaviour change theory, to assess the current landscape of Australian physicians' genomic medicine practice, perceptions of proximity and individual preparedness, and preferred models of practice and continuing education. The survey was advertised nationally through 10 medical colleges, 24 societies, 62 hospitals, social media, professional networks and snowballing. 409 medical specialists across Australia responded, representing 30 specialties (majority paediatricians, 20%), from mainly public hospitals (70%) in metropolitan areas (75%). Half (53%) had contacted their local genetics services and half (54%) had ordered or referred for a gene panel or exome/genome sequencing test in the last year. Two-thirds (67%) think genomics will soon impact their practice, with a significant preference for models that involved genetics services (p<0.0001). Currently, respondents mainly perform tasks associated with pretest family history taking and counselling, but more respondents expect to perform tasks at all stages of testing in the future, including tasks related to the test itself, and reporting results. While one-third (34%) recently completed education in genomics, only a quarter (25%) felt prepared to practise. Specialists would like (more) education, particularly on genomic technologies and clinical utility, and prefer this to be through varied educational strategies. This survey provides data from a breadth of physician specialties that can inform models of genetic service delivery and genomics education. The findings support education providers designing and delivering education that best meet learner needs to build a competent, genomic-literate workforce. Further analyses are underway to characterise early adopters of genomic medicine to inform strategies to increase engagement.


Accounting of the Use and Emissions of Polychlorinated Biphenyl Compounds (PCBs) in India, 1951-2100.
Polychlorinated biphenyl compounds (PCBs) are highly toxic organic chemicals still prevalent in the environment. While global inventories of the use and emissions of PCBs have been developed, estimates for individual countries determined using bottom-up approaches are few and often show different trends from the global inventory. Here, we determine the past, present, and future consumption and emissions of PCBs in India. A mass balance model was used to estimate middle (low-high) emissions in the period 1950-2100. Up to 7296 tonnes of PCBs have been used in transformers. PCBs imported as wastes are estimated to be approximately 5000 (2400-9100) tonnes. Total emissions from the use and disposal of transformers, industrial processes, and imported waste disposal are estimated to become 13 (0.1-537) tonnes, 89.26 (0.5-178) tonnes, 63 (3-910) tonnes, respectively, in the period 1950-2100. Congener-specific emissions are relatively high for low-chlorinated PCBs (-8, 18, 28, 31, 52, 101, 110, 118, 153, range: 0.1-118 tonnes). We find that industrial emissions are becoming important sources of PCBs and may become predominant, depending on emission scenarios.


[Use of Nordic walking in the complex treatment of lower urinary tract symptoms in chronic prostatitis].
Lower urinary tract symptoms (LUTS) caused by chronic prostatitis (CP) are a common condition. Standard antibiotic therapy is often not justified, since bacterial prostatitis is presented in up to 10% of the cases, while there is no effect on the risk factors for the development and relapse of the disease. This study is aimed at one of the main risk factors for LUTS - a sedentary lifestyle. To increase the effectiveness of treatment for LUTS in men using the Nordic Walking technique. 84 patients with CP were examined, which were divided into 3 groups. The 1st group (control) included 32 patients who received standard antibacterial therapy. In the 2nd group (primary) - 32 patients who received complex therapy (antibiotic therapy + 4-week course of Nordic walking). The third group (comparison) included 20 patients who were prescribed monotherapy - Nordic walking for 4 weeks. The dynamics of clinical manifestations was assessed using CP Symptom Scale (NIH-CPSI) and the international symptom index for diseases of the prostate IPSS; the volume of residual urine, uroflowmetric data, and the level of leukocytes in the secretion of the prostate gland were estimated. Laboratory studies showed a significantly faster decrease in the number of leukocytes in the secretion of the prostate gland on the 7th and 14th day in the primary (2nd) group (p<0.001). Clinical symptoms according to the NIH-CPSI and IPSS scales significantly regressed in the primary group on the 14th and 28th days of therapy. Uroflowmetric data and the volume of residual urine improved more significantly in the primary group by the 14th and 28th days. Patients receiving combined therapy showed significantly better treatment results in the form of faster normalization of prostate secretion, regression of clinical symptoms, improved urine flow rate and a decrease in residual urine volume after 4 weeks of therapy. When choosing management algorithm for patients with LUTS, it is advisable to use an integrated approach: in addition to antibiotic therapy, Nordic walking is recommended 3 times a week for at least 4 weeks.


Effects of a high-monounsaturated fat diet on glucose and lipid metabolisms in normal and diabetic mice.
The beneficial effects of high-monounsaturated fat (high-MUFA) diets on diabetic patients have been reported, whereas studies concerning the effects on animals have been few. Although experiments on animals should be useful in elucidating underlying mechanisms, it is not clear even whether there are benefits of a high-MUFA diet in animals. This study examined the short-term effects of a high-MUFA diet on normal and genetically diabetic mice. The high-MUFA diet supplied 38% of the total calories as fat (26% from MUFA), while a regular diet was 13% fat (3% from MUFA). Normal C5 7BL/6J and diabetic C57BL/KsJ-db/db mice were fed either the regular or the high-MUFA diet for 1 wk. Serum glucose and lipid levels were then measured. In normal mice, hepatic triglyceride production was also compared between the two dietary groups using the Triton WR1339 method. An oral glucose tolerance test was conducted on the diabetic mice. After 1 wk of feeding to normal mice, the high-MUFA diet was seen to lower serum triglyceride levels and reduce hepatic triglyceride production in comparison with the regular diet; it is suggested that the lowering of triglyceride consists of mechanisms including reduced hepatic triglyceride production. When diabetic mice were fed the high-MUFA diet with a controlled caloric intake, the serum glucose levels lowered without an accompanying deterioration in lipid metabolism and the impaired glucose tolerance was ameliorated. This study demonstrates that a high-MUFA diet can lower serum triglyceride levels in normal mice and improve disorders of glucose metabolism in diabetic mice.


Prediction of glomerular filtration rate by serum creatinine and beta 2-microglobulin.
The reciprocal of serum creatinine concentration (1/Cr) is often used to predict glomerular filtration rate (GFR). Serum creatinine also varies with age, size, and muscle mass, and so it may inaccurately estimate GFR. The reciprocal of serum beta 2-microglobulin (1/beta 2mu) has been proposed as an alternative estimator of GFR. This study compares 1/Cr and 1/beta 2mu as predictors of GFR as measured by 125I-iothalamate clearance (CIOT) and creatinine clearance (CCr) in 29 subjects with a wide range of age, size and kidney function, and including 12 chronic hemodialysis patients. 1/beta 2mu was a better predictor of CIOT (r=0.90) and CCr (r=0.87) than 1/Cr (r=0.50 and 0.78) was. In fact, in the nondialysis population, 1/beta 2mu predicted CIOT (r=0.86) about as well as CCr predicted CIOT (r=0.87). Beta 2mu was less dependent on body size than Cr and unlike Cr, was not influenced by dialysis and did increase as GFR decreased with age. Beta 2mu can be useful as an alternative clinical estimate of GFR, particularly when Cr is considerably influenced by factors other than renal function.


Admissions for chronic ambulatory care sensitive conditions - a useful measure of potentially preventable admission?
Potentially preventable hospital admission (an admission deemed to be potentially preventable given appropriate care in the community-based healthcare setting) has been a topic of international research attention for almost three decades. Recently this has been largely driven by the imperative to reduce ever-increasing unplanned hospital admissions. However, identifying potentially preventable admissions is difficult. As a result, the population level indicator of admissions for ambulatory care sensitive conditions (ACSCs) has been used as a proxy measure for potentially preventable admission. The adoption of this measure has become common, and in Australia, the rate of admissions for chronic ACSCs is now an important component of measuring health system performance and accountability, and is directly linked to funding. Admission for a chronic ACSC is also used to identify individuals for targeting of interventions to reduce preventable admissions. Hospital admission for chronic ACSCs is a population measure based on admission diagnoses, it therefore should not be used to identify individual preventable admissions. At present we are unable to determine individual admissions that are deemed to be preventable or, therefore, articulate the factors associated with admissions which are preventable. As we are currently unable to identify individual admissions that are preventable, little is understood about the underlying causes and factors contributing to preventable admissions. A means of assessing preventability of individual admissions is required. Only then can we explore the antecedents, and patient and clinician perspectives on preventable admissions. Until we have a clearer understanding of this, our capacity to inform policy and program development remains compromised.


An unusual complication of acute myocardial infarction.
Right ventricular infarction occurs exclusively as a complication of a transmural inferoseptal left ventricular infarction. Up to 50% of patients with transmural inferoseptal infarction may have associated right ventricular involvement. In the majority of cases, however, the right ventricular involvement is clinically silent. However, significant right ventricular infarction is being recognized with greater frequency. Clinically, distended neck veins and hypotension are frequent findings in acute right ventricular infarction and their presence should suggest the diagnosis. Damage to the right ventricle causes poor contractility and output, and thus the right ventricle functions more as a passive conduit to the left ventricle. The patient should lie with the bed flat to improve volume return to the right and left ventricles. Volume expansion is used when left ventricular filling pressure is low and there is clinical evidence of poor peripheral perfusion.


Regio- and stereo-selective hydroxylation of abietic acid derivatives by Mucor circinelloides and Mortierella isabellina.
Mucor circinelloides and Mortierella isabellina hydroxylated dehydroabietic acid (DehA). DehA was converted regio- and stereo-selectively by whole cells of Mr. circinelloides to give 2alpha-hydroxydehydroabietic acid in a 75% molar conversion yield (11 mM from 14.7 mM DehA) after 72 h in the cultivation medium containing 3% (v/v) Tween 80. With cells of Ma. isabellina, under the same conditions, 20.5 mM (6.5 g l(-1)) 2-hydroxydehydroabietic acid (alpha/beta=81/19) was formed from 26.4 mM DehA.


Epidemiology of human atopic dermatitis--seven areas of notable progress and seven areas of notable ignorance.
This narrative review highlights areas within the epidemiology of human atopic dermatitis (AD) where significant progress has been made and where considerable ignorance still exists. The review is supported by systematic reviews wherever possible, with the purpose of stimulating fresh approaches to human and veterinary research into AD. Areas of progress include valid and repeatable methods of disease definition, global documentation of disease prevalence and impact, clarification of the role of some genetic factors, such as filaggrin gene mutations, clear evidence that environmental factors are key, as demonstrated by the positive social class gradient and rising prevalence, a possible protective effect of infections in early life, documentation of comorbidities, such as a reduced risk of glioma, and mapping the evidence base through systematic reviews and an online global resource of clinical trials. Areas where significant uncertainty still exists include the question of whether AD is more than one disease, the tendency for researchers to look at the same old risk factors, lack of specific environmental risk factors that are amenable to manipulation, inconsistencies in the hygiene hypothesis, sparse knowledge about adult AD, lack of evidence that eczema can be prevented, and little scientific work exploring what causes flares in people with established AD.


Toward understanding and treating violence in America: some contributions from group dynamic and group therapy perspectives: introduction to part I.
The co-editors introduce a two-part Special Section of the Journal devoted to understanding and treating violence in America. They examine the relevance of clinical experience for contributions that can be made by group therapists and group dynamic thinkers to the growing national dialogue about this problem. The pervasive nature, causes, and different forms of violence in the United States are compared with those found in other countries. Underlying sociocultural values and myths, historical and current cultural contexts are considered breeding grounds for potential violence. How therapists can promote healthy change in their groups and in the broader society is explored. The articles contained in part one are reviewed against this backdrop.


Novel IL-2-Poly(HPMA)Nanoconjugate Based Immunotherapy.
Interleukin-2 (IL-2) possesses a strong stimulatory activity for activated T and NK cells and it is an attractive molecule for immunotherapy. Nevertheless, extremely short half-life and severe toxicities associated with high-dose IL-2 treatment are serious and limiting drawbacks. In order to increase IL-2 half-life in vivo, we covalently conjugated synthetic semitelechelic polymeric carrier based on N-(2-hydroxypropyl)methacrylamide (HPMA) to IL-2. Thus, we synthesized IL-2-poly(HPMA) conjugate containing 2-3 polymer chains per IL-2 molecule in average. Such conjugate has lower biologic activity in comparison to IL-2 in vitro. However, it exerts much higher activity than IL-2 in vivo as shown by expansion of memory CD8+ T, NK, NKT, γδT and Treg cells. Moreover, IL-2-poly(HPMA) extremely effectively potentiates CD8+ T cell peptide-based vaccination. IL-2-poly(HPMA) shows also much longer half-time in circulation than IL-2 (-4 h versus -5 min). Collectively, modification of IL-2 with poly(HPMA) chains dramatically improves its potency and pharmacologic features in vivo, which have implications for immunotherapy. To our knowledge, this is the first proof-of-concept report of the use of polymer/protein modification of IL-2 to obtain more pronounced biological activity.


Submental liposuction extraction with hard chin augmentation.
Liposuction techniques utilizing fat extraction sculpturing of the neck and simultaneous chin augmentation are described herein. In young persons, extraction is carried out in the submental areas. In older patients, who have diffuse fat accumulations and less skin tone, extraction includes the submental and submandibular areas, along with dry tunneling to the lateral borders of the sternocleidomastoid muscle and inferiorly to the suprasternal notch. When the skin tone is poor, with excessive redundancy, combined rhytidectomy with both open and closed liposuction is performed. With liposuction submental sculpturing , necessity for chin augmentation is critical to tauten the submental skin and to give a better neck-chin angle. Hard chin implants are preferred over the soft gelatinous types. We utilized a hard acrylic implant ( Rish ) in all cases. Results obtained by the dual approach of submental fat suction sculpturing and hard chin augmentation have been excellent.


Accuracy Evaluation of a 3D Ultrasound-guided Biopsy System.
Early detection of prostate cancer is critical in maximizing the probability of successful treatment. Current systematic biopsy approach takes 12 or more randomly distributed core tissue samples within the prostate and can have a high potential, especially with early disease, for a false negative diagnosis. The purpose of this study is to determine the accuracy of a 3D ultrasound-guided biopsy system. Testing was conducted on prostate phantoms created from an agar mixture which had embedded markers. The phantoms were scanned and the 3D ultrasound system was used to direct the biopsy. Each phantom was analyzed with a CT scan to obtain needle deflection measurements. The deflection experienced throughout the biopsy process was dependent on the depth of the biopsy target. The results for markers at a depth of less than 20 mm, 20-30 mm, and greater than 30 mm were 3.3 mm, 4.7 mm, and 6.2 mm, respectively. This measurement encapsulates the entire biopsy process, from the scanning of the phantom to the firing of the biopsy needle. Increased depth of the biopsy target caused a greater deflection from the intended path in most cases which was due to an angular incidence of the biopsy needle. Although some deflection was present, this system exhibits a clear advantage in the targeted biopsy of prostate cancer and has the potential to reduce the number of false negative biopsies for large lesions.


Case report: polysomnographic effects of thalamotomy for torsion dystonia.
We report a patient with torsion dystonia whose polysomnographic recordings revealed poor sleep and a pronounced and almost continuous type of spindle activity during non-rapid eye movement sleep. Rapid eye movement sleep was also reduced. These changes proved to be independent of medications. After a clinically successful unilateral thalamic operation, a normalization of sleep parameters and a reduction of the high amplitude sleep spindles was observed, implying that the regulation of sleep spindles and the advancement of dystonic symptoms are affected by a common mechanism.


Psychological aspects of juvenile fibromyalgia syndrome: a literature review.
Juvenile fibromyalgia syndrome (JFMS) is a non-inflammatory chronic pain condition that occurs mainly in girls aged 9-15 years. JFMS is characterized by constant widespread pain in different parts of the body, poor sleep quality, daytime sleepiness and an altered mood. Concomitant psychological and organic factors result in a diminished capacity to cope with pain. The quality of life of individuals with chronic pain and their caregivers is severely restricted and the occurrence of symptoms of anxiety and depression is common in this population. The aim of the present study was to perform a systematic review of the literature on psychosocial factors related to JFMS. The findings reveal differences in opinion between patients and family members regarding the effect of the condition, as mothers tend to classify JFMS as more severe than the patients themselves. Individuals with JFMS seem to share the same personality traits and there seems to be a type of family environment that is favorable to the occurrence of this condition. Psychological and functional aspects should be treated with methods that can help patients and family members alter their coping strategies regarding day-to-day problems, attenuate the dysfunctional consequences of pain and fatigue and diminish the risk of catastrophizing that individuals submitted to constant pain develop in relation to their surrounding environment.


Primary hyperparathyroidism is associated with increased circulating bone marrow-derived progenitor cells.
Recently, parathyroid hormone (PTH) was shown to support survival of progenitor cells in bone marrow. The release of progenitor cells occurs in physiological and pathological conditions and was shown to contribute to neovascularization in tumors and ischemic tissues. In the present study we sought to investigate prospectively the effect of primary hyperparathyroidism (PHPT) on mobilization of bone marrow-derived progenitor cells. In 22 patients with PHPT and 10 controls, defined subpopulations of circulating bone marrow-derived progenitor cells (BMCs) were analyzed by flow cytometry (CD45(+)/CD34(+)/CD31(+) cells indicating endothelial progenitor cells, CD45(+)/CD34(+)/c-kit(+) cells indicating hematopoietic stem cells, and CD45(+)/CD34(+)/CXCR4(+) cells indicating progenitor cells with the homing receptor CXCR4). Cytokine serum levels (SCF, SDF-1, VEGF, EPO, and G-CSF) were assessed using ELISA. Levels of PTH and thyroid hormone as well as serum electrolytes, renal and liver parameters, and blood count were analyzed. Our data show for the first time a significant increase of circulating BMCs and an upregulation of SDF-1 and VEGF serum levels in patients with PHPT. The number of circulating BMCs returned to control levels measured 16.7 +/- 2.3 mo after surgery. There was a positive correlation of PTH levels with the number of CD45(+)/CD34(+)/CD31(+), CD45(+)/CD34(+)/c-kit(+), and CD45(+)/CD34(+)/CXCR4(+) cells. However, there was no correlation between cytokine serum concentrations (SDF-1, VEGF) and circulating BMCs. Serum levels of G-CSF, EPO, and SCF known to mobilize BMCs were even decreased or remained unchanged, suggesting a direct effect of PTH on stem cell mobilization. Our data suggest a new function of PTH mobilizing BMCs into peripheral blood.


The use of self-management strategies by people with rheumatoid arthritis.
To investigate seven common arthritis self-management methods used by people with rheumatoid arthritis (RA) by studying their frequency of use and the patients' belief in their benefits. Also to look at how people obtained information about such methods. Cross-sectional survey. Forty-one people with RA attending a rheumatology outpatient department at a large district general hospital. Twenty-three recalled receiving some self-management advice from rheumatology team members, but most stated the commonest source of information was arthritis books and leaflets (n = 29). Two-thirds used technical aids and rest on a daily basis, half used exercise and heat (half doing so daily). A quarter to a half used relaxation, joint protection and working splints (half doing so daily). People with RA reported using three or four self-management methods simultaneously to help control symptoms. In the main, believing a method to be beneficial was strongly related to its use. However, exercise and joint protection, whilst widely believed to be beneficial, were less used. People reported problems with knowing how to do these correctly, having insufficient time to practise sufficiently and difficulty in establishing habits.


Bi-allelic mutations of LONP1 encoding the mitochondrial LonP1 protease cause pyruvate dehydrogenase deficiency and profound neurodegeneration with progressive cerebellar atrophy.
LonP1 is crucial for maintaining mitochondrial proteostasis and mitigating cell stress. We identified a novel homozygous missense LONP1 variant, c.2282 C > T, (p.Pro761Leu), by whole-exome and Sanger sequencing in two siblings born to healthy consanguineous parents. Both siblings presented with stepwise regression during infancy, profound hypotonia and muscle weakness, severe intellectual disability and progressive cerebellar atrophy on brain imaging. Muscle biopsy revealed the absence of ragged-red fibers, however, scattered cytochrome c oxidase-negative staining and electron dense mitochondrial inclusions were observed. Primary cultured fibroblasts from the siblings showed normal levels of mtDNA and mitochondrial transcripts, and normal activities of oxidative phosphorylation complexes I through V. Interestingly, fibroblasts of both siblings showed glucose-repressed oxygen consumption compared to their mother, whereas galactose and palmitic acid utilization were similar. Notably, the siblings' fibroblasts had reduced pyruvate dehydrogenase (PDH) activity and elevated intracellular lactate:pyruvate ratios, whereas plasma ratios were normal. We demonstrated that in the siblings' fibroblasts, PDH dysfunction was caused by increased levels of the phosphorylated E1α subunit of PDH, which inhibits enzyme activity. Blocking E1α phosphorylation activated PDH and reduced intracellular lactate concentrations. In addition, overexpressing wild-type LonP1 in the siblings' fibroblasts down-regulated phosphoE1α. Furthermore, in vitro studies demonstrated that purified LonP1-P761L failed to degrade phosphorylated E1α, in contrast to wild-type LonP1. We propose a novel mechanism whereby homozygous expression of the LonP1-P761L variant leads to PDH deficiency and energy metabolism dysfunction, which promotes severe neurologic impairment and neurodegeneration.


Alternaria malicola sp. nov., a New Pathogen Causing Fruit Spot on Apple in China.
Alternaria spp. are pathogens of several diseases that pose significant threats to apple production. Several putative Alternaria sp. isolates were obtained from lesions of a disease commonly referred to as black dot on apple fruit in Shaanxi Province, China. Pathogenicity tests using mycelial plugs and conidial suspensions indicated that this isolate could cause leaf blotch, as well as moldy core and black dot on fruit. On the basis of sequence analysis of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), RNA polymerase second largest subunit, and translation elongation factor 1-α, an isolate clustered with the Alternaria sect. Ulocladioides. By combining GAPDH, major allergen Alta1, mating type protein 1-2-1, and the AGA1 gene sequence analysis and morphological description, the isolates were identified as a new species named Alternaria malicola. Our finding expands the documented diversity of apple pathogens within the genus Alternaria and clarifies the taxonomy of the pathogen assemblage that may be associated with three apple diseases.


Molecular imaging of p53 signal pathway in lung cancer cell cycle arrest induced by cisplatin.
Cisplatin is a commonly employed chemotherapy drug for lung malignancy. However its efficacy is limited by acquired drug resistance and lacking of an in vivo real-time monitoring approach. The aim of this study is to investigate the effect of cisplatin on lung adenocarcinoma cell line p53-RE-Fluc/A549 in vivo via non-invasive reporter gene by molecular imaging. For this study, we employed p53-RE-Fluc/A549 cells that overexpressed a vector with three tandem repeats of p53 response element followed by the luciferase reporter gene. P53 activity was evaluated by optical imaging and verified by Western blot after cells were exposed to 10 µM cisplatin for 72 h. The cell cycle was mainly blocked at the S- and G2/M-phases after cisplatin treatment, whereas no significant change was observed in cell apoptotic index. Increased expression of p21 and Bcl-2 as well as decreased expression of Bax were observed after cisplatin treatment by Western blotting. Longitudinal in vivo bioluminescent imaging (BLI) revealed that the p53 activity was increased from 24 to 48 h after transient cisplatin treatment in p53-RE-Fluc/A549-bearing nude mice. RNA sequencing further revealed that cell cycle and p53 signaling pathway genes, such as E2F1, CCNA2, CDK1, and CCNE2 were significantly downregulated after long-term cisplatin treatment. Thus, our study showed that cisplatin exerts its cytotoxic effect through blockage of the cell cycle and may be partly regulated by the p53 signaling pathway. Furthermore, molecular imaging is a useful tool to investigate the mechanism and evaluate the effect of chemotherapy drugs both in vivo and in vitro.


Multiple bloodstream infections in pediatric stem cell transplant recipients: A case series.
Bacterial bloodstream infections (BSIs) are associated with poor outcomes following stem cell transplantation (SCT). We describe the demographics, treatment, complications, and outcome of 23 pediatric SCT recipients who developed three or more BSIs in the first year after SCT at our center from 2011 through 2016. The majority underwent allogeneic SCT (n = 22/23;96%), mainly from an unrelated donor (n = 19/22,86%); developed grade 2-4 graft versus host disease (GVHD; n = 14/23, 61%), all steroid refractory; and were diagnosed with thrombotic microangiopathy (n = 21/23, 91%). One-year overall survival was 56% (n = 13/23). We observed a high rate of transplant-associated thrombotic microangiopathy and steroid-refractory acute GVHD in patients with three or more BSIs.


Advocating for Automatic Eligibility for Early Intervention Services for Children Exposed to Lead.
Lead poisoning remains one of the most prevalent and preventable environmental health hazards affecting children. Low-level lead exposure has been shown to significantly increase the risk of learning disabilities, behavioral problems, and developmental delay. It also has substantial social implications as lead toxicity disproportionately affects children from low-income, ethnic minority households. Early Intervention (EI) is a state-based federal program (Part C of the Federal Individuals with Disabilities Education Act) created to support the development of children from birth to age 3 years who are experiencing or who are at risk for developmental delay. In 2016, an Illinois EI and Lead Workgroup was established to address the best practice of offering EI services to children who have been exposed to lead. This work serves as a template for advocates and health providers in other states to establish automatic eligibility for EI services at low levels of lead exposure, while also demonstrating the capacity to serve these children without overwhelming the state's EI system. [Pediatr Ann. 2018;47(10):e413-e418.].


Conversion of Bacillus subtilis OhrR from a 1-Cys to a 2-Cys peroxide sensor.
OhrR proteins can be divided into two groups based on their inactivation mechanism: 1-Cys (represented by Bacillus subtilis OhrR) and 2-Cys (represented by Xanthomonas campestris OhrR). A conserved cysteine residue near the amino terminus is present in both groups of proteins and is initially oxidized to the sulfenic acid. The B. subtilis 1-Cys OhrR protein is subsequently inactivated by formation of a mixed-disulfide bond with low-molecular-weight thiols or by cysteine overoxidation to sulfinic and sulfonic acids. In contrast, the X. campestris 2-Cys OhrR is inactivated when the initially oxidized cysteine sulfenate forms an intersubunit disulfide bond with a second Cys residue from the other subunit of the protein dimer. Here, we demonstrate that the 1-Cys B. subtilis OhrR can be converted into a 2-Cys OhrR by introducing another cysteine residue in either position 120 or position 124. Like the X. campestris OhrR protein, these mutants (G120C and Q124C) are inactivated by intermolecular disulfide bond formation. Analysis of oxidized 2-Cys variants both in vivo and in vitro indicates that intersubunit disulfide bond formation can occur simultaneously at both active sites in the protein dimer. Rapid formation of intersubunit disulfide bonds protects OhrR against irreversible overoxidation in the presence of strong oxidants much more efficiently than do the endogenous low-molecular-weight thiols.


STING is a cell-intrinsic metabolic checkpoint restricting aerobic glycolysis by targeting HK2.
Evasion of antitumour immunity is a hallmark of cancer. STING, a putative innate immune signalling adaptor, has a pivotal role in mounting antitumour immunity by coordinating innate sensing and adaptive immune surveillance in myeloid cells. STING is markedly silenced in various human malignancies and acts as a cell-intrinsic tumour suppressor. How STING exerts intrinsic antitumour activity remains unclear. Here, we report that STING restricts aerobic glycolysis independent of its innate immune function. Mechanistically, STING targets hexokinase II (HK2) to block its hexokinase activity. As such, STING inhibits HK2 to restrict tumour aerobic glycolysis and promote antitumour immunity in vivo. In human colorectal carcinoma samples, lactate, which can be used as a surrogate for aerobic glycolysis, is negatively correlated with STING expression level and antitumour immunity. Taken together, this study reveals that STING functions as a cell-intrinsic metabolic checkpoint that restricts aerobic glycolysis to promote antitumour immunity. These findings have important implications for the development of STING-based therapeutic modalities to improve antitumour immunotherapy.


Can resonance frequency analysis detect narrow marginal bone defects around dental implants? An ex vivo animal pilot study.
Resonance frequency analysis (RFA) is applied to assess implant stability, as expressed by the implant stability quotient (ISQ). This study aimed to investigate the potential of RFA devices to identify narrow marginal bone defects around implants. Twenty-eight Straumann bone level (BL) implants and 28 bone level tapered (BLT) implants were placed ex vivo in porcine ribs. Implants in the control group (A) were fully submerged in the bone. In three experimental groups, implants were placed with a 0.9-mm circumferential marginal bone defect extending 2 mm (B), 4 mm (C) and 6 mm (D) apically. Two RFA devices were used to measure implant stability. ISQ values decreased as the defects' depth increased, with the greatest reduction observed between full bone (A) and 2-mm defects (B) (P < 0.001). No significant differences were found in the ISQ values recorded from BL and BLT implants. ISQ values can effectively detect narrow, intrabony marginal bone defects, in particular when involving the first coronal 2 mm. This finding could have implications for the early diagnosis of conditions affecting the marginal bone, such as peri-implantitis. Further research is required to investigate if such findings can be replicated after osseointegration is achieved.


Evaluation of recombinant proteins of Burkholderia mallei for serodiagnosis of glanders.
Glanders is a contagious disease caused by the Gram-negative bacillus Burkholderia mallei. The number of equine glanders outbreaks has increased steadily during the last decade. The disease must be reported to the Office International des Epizooties, Paris, France. Glanders serodiagnosis is hampered by the considerable number of false positives and negatives of the internationally prescribed tests. The major problem leading to the low sensitivity and specificity of the complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e., crude preparations of whole cells. False-positive results obtained from other diagnostic tests utilizing crude antigens lead to financial losses to animal owners, and false-negative results can turn a risk into a possible threat. In this study, we report on the identification of diagnostic targets using bioinformatics tools for serodiagnosis of glanders. The identified gene sequences were cloned and expressed as recombinant proteins. The purified recombinant proteins of B. mallei were used in an indirect ELISA format for serodiagnosis of glanders. Two recombinant proteins, 0375H and 0375TH, exhibited 100% sensitivity and specificity for glanders diagnosis. The proteins also did not cross-react with sera from patients with the closely related disease melioidosis. The results of this investigation highlight the potential of recombinant 0375H and 0375TH proteins in specific and sensitive diagnosis of glanders.


Peripartal alterations of calcitonin gene-related peptide and minerals in dairy cows affected by milk fever.
Milk fever, a metabolic disease of dairy cattle, is associated with perturbations of calcium homeostasis, the pathogenesis of which is not yet completely understood. The aim of this study was to investigate plasma concentrations of calcitonin gene-related peptide and selected minerals and metabolites in periparturient cows with and without milk fever. Plasma concentrations of calcitonin gene-related peptide, as well as calcium, phosphate, magnesium, iron, glucose, lactate, and cortisol, were determined in multiple plasma samples from Jersey cows with and without spontaneous milk fever. Cows affected by milk fever (n = 5) had lower concentrations of calcitonin gene-related peptide (P = .038) and inorganic phosphate (P < .001) in plasma than did the controls (n = 5). Also, these cows tended to have lower calcium concentrations (P = .071). Magnesium, iron, lactate, glucose, and cortisol concentrations were comparable between both groups of cows (P > .10). Around the day of calving, plasma concentrations of lactate, glucose, and cortisol increased and the concentration of iron decreased in all cows (P ≤ .01). Despite the limited number of cows evaluated, this report is the first to indicate lowered concentrations of calcitonin gene-related peptide as part of the metabolic changes during milk fever in cows. Further work with a larger cohort of animals is warranted to understand the precise role of calcitonin gene-related peptide and the potential associations with disturbances in plasma minerals typically observed during milk fever.


Ratiometric Visualization of NO/H2S Cross-Talk in Living Cells and Tissues Using a Nitroxyl-Responsive Two-Photon Fluorescence Probe.
It is of scientific significance to explore the intricate relationship between two crucial gasotransmitters nitric oxide (NO) and hydrogen sulfide (H2S) because they exert similar and interdependent biological actions within the living organisms. Nevertheless, visualization of the NO/H2S crosstalk using effective molecular imaging tools remains challenging. To address this issue, and given that nitroxyl (HNO) has been implicated as the interdependent production of NO and H2S via a network of cascading chemical reactions, we herein design a ratiometric two-photon fluorescent probe for HNO, termed TP-Rho-HNO, which consists of benzo[h]chromene-rhodol scaffold as two-photon energy transfer cassette with phosphine moiety as specific HNO recognition unit. The newly proposed probe has been successfully applied in ratiometric two-photon bioimaging of endogenous HNO derived from NO and H2S interaction in the human umbilical vein cells (HUVECs) and as well as in rat brain tissues. Intriguingly, the imaging results consistently demonstrate that the mutually dependent upgeneration of H2S and NO are present in living biosystems, indicating that this molecular probe would provide a powerful approach to elucidate the chemical foundation for the anfractuous cross-talk between the NO and H2S signaling pathways in biology.


Replacement equivalence of H- and argon in small (Ar)nH- clusters from optimized structure calculations.
The structural properties of some of the smaller ionic clusters of argon atoms containing the atomic impurity H-, ArnH- with n from 2 up to 7, are examined using different modeling for the interactions within each cluster and by employing different theoretical treatments, both classical and quantum, for the energetics. The same calculations are also carried out for the corresponding neutral homogeneous clusters Ar(n+1). The results of the calculations, the physical reliability of the interactions modeling, and the similarities and the difference between the anionic and the neutral complexes are discussed in some detail. The emerging picture shows that, due to specific features of the employed atom-atom potentials, the ArnH- and Ar(n+1) clusters present very similar structures, where the H- dopant substitutes for one of the outer Ar atoms but does not undergo as yet solvation within such small clusters.


To Be Seen, Heard, and Valued: Strategies to Promote a Sense of Belonging for Women and Underrepresented in Medicine Physicians.
Lingering unconscious biases and daily cues continue to permeate and persist in academic medicine environments in the form of the exclusion of physicians who are women or racially/ethnically underrepresented in medicine. Academic medicine environments must change so that women and underrepresented in medicine racial/ethnic groups are seen, heard, and valued. A shared awareness among faculty, administrators, and trainees can inform the development of intentional strategies to alter individual behaviors, academic spaces, and institutional processes to cultivate a sense of belonging. Shifting the norms in medicine and the course of historical exclusion will require professional development in areas of inclusive teaching practices, skills to cultivate mentoring relationships with diverse trainees, and fostering discussions about the relevance of personal identity, as well as attention to the symbolism and imagery in institutional messages (e.g., portraits on the walls, website, marketing campaigns) and to the value of including community involvement in productivity metrics.


[Technical problems in the preparation of biopsy samples of the spleen].
Spleen tissue sampling for bioptical examination should be performed after mutual consultation of clinician and pathologist. Clinician should know what is expected from the pathologist. This can influence preparation and sampling method considerably. In a great amount of cases the whole organ is removed. A lab technician is sampling on her own and her contribution to a good result can be either positive or negative. Imprint preparations from the tissue sections can be useful. Specialized examination should be performed in the presence of a pathologist or his co-workers. The active approach of a lab technician can significantly contribute to the value of examined sample of the spleen, of the organ until recently considered to be not important or full of mysteries.


Improved immunocytochemical staining of carcinogen-DNA adducts by a capillary slot block system.
We developed an immunocytochemical protocol in which incubation occurs in a capillary slot instead of the conventional horizontal drop. Slots of constant width were formed by placing slides on top of each other with parafilm spacer layers in between. Cryostat or semi-thin plastic-embedded sections were cut from organs of carcinogen-treated experimental animals. Carcinogen-DNA adducts were visualized in the affected nuclei by a double peroxidase-antiperoxidase method using rabbit antisera specific for certain DNA adducts formed. The staining in capillary slot blocks offered better staining reproducibility than the conventional method. This is particularly important when the staining intensity must be quantified. In addition, handling of the blocks was substantially less laborious than the individual treatment of slides, making this protocol especially suitable for larger series of slides. Other applications for the capillary slot block protocol should be enzyme histochemistry and in situ hybridization.


Equine placentation.
A tough, elastic glycoprotein capsule envelops the equine blastocyst between Days 6 and 23 after ovulation. It maintains the spherical configuration of, and provides physical support for, the embryo as it traverses the entire uterine lumen during Days 6-17, propelled by myometrial contractions that are stimulated by pulsatile release of prostaglandin F2alpha and prostaglandin E2. The capsule also accumulates constituents of the exocrine secretions of the endometrial glands ('uterine milk') as nutrients for the mobile embryo as it releases its antiluteolytic maternal recognition-of-pregnancy signal to the whole of the surface of the endometrium. Mobility ceases abruptly on Day 17 with a sudden increase in uterine tonicity that 'fixes' the conceptus at the base of one of the uterine horns. At Day 35, the trophoblast of the spherical conceptus has separated into its invasive and non-invasive components. The former, distinguished as the thickened, annulate chorionic girdle, invades the maternal endometrium to form the unique endometrial cups. These secrete a chorionic gonadotrophin that synergizes with pituitary follicle-stimulating hormone to induce secondary luteal development in the maternal ovaries. The cup cells express foreign fetal antigens that stimulate strong maternal humoral and cell-mediated immune responses, which curtail their lifespan. The non-invasive trophoblast of the allantochorion establishes a stable microvillous contact with the endometrial epithelium around Day 40 and, over the next 100 days, develops a complex multibranched interdigitation with the endometrium to form the microcotyledonary haemotrophic exchange units that cover the entire surface of the diffuse epitheliochorial placenta. Reduction in the effective total area of fetomaternal contact at this placental interface, by competition between twin conceptuses for the limited area of available endometrium, by attachment of the allantochorion to an imperfect endometrium in a mare with endometrosis, or following cross-breeding or embryo transfer between a sire and dam of dissimilar size, will all induce intrauterine growth retardation of the fetus and runting of the foal, which persists into adult life. Over 40 years ago, Professor Roger Short and his colleagues determined that the high concentrations of conventional and unique ring B unsaturated oestrogens in the blood and urine of mares during the second half of pregnancy stem from placental aromatization of large quantities of C-19 precursor molecules secreted by the temporarily hypertrophic fetal gonads. Placental production of progesterone and 5alpha-reduced progestagens, on the other hand, depends on both maternal and fetal adrenal sources of pregnenelone.


Oral treatment with Eubacterium hallii improves insulin sensitivity in db/db mice.
An altered intestinal microbiota composition is associated with insulin resistance and type 2 diabetes mellitus. We previously identified increased intestinal levels of Eubacterium hallii, an anaerobic bacterium belonging to the butyrate-producing Lachnospiraceae family, in metabolic syndrome subjects who received a faecal transplant from a lean donor. To further assess the effects of E. hallii on insulin sensitivity, we orally treated obese and diabetic db/db mice with alive E. hallii and glycerol or heat-inactive E. hallii as control. Insulin tolerance tests and hyperinsulinemic-euglycemic clamp experiments revealed that alive E. hallii treatment improved insulin sensitivity compared control treatment. In addition, E. hallii treatment increased energy expenditure in db/db mice. Active E. hallii treatment was found to increase faecal butyrate concentrations and to modify bile acid metabolism compared with heat-inactivated controls. Our data suggest that E. hallii administration potentially alters the function of the intestinal microbiome and that microbial metabolites may contribute to the improved metabolic phenotype.


Sending and Receiving Hedgehog Signals.
Communication between cells pervades the development and physiology of metazoans. In animals, this process is carried out by a relatively small number of signaling pathways, each consisting of a chain of biochemical events through which extracellular stimuli control the behavior of target cells. One such signaling system is the Hedgehog pathway, which is crucial in embryogenesis and is implicated in many birth defects and cancers. Although Hedgehog pathway components were identified by genetic analysis more than a decade ago, our understanding of the molecular mechanisms of signaling is far from complete. In this review, we focus on the biochemistry and cell biology of the Hedgehog pathway. We examine the unique biosynthesis of the Hedgehog ligand, its specialized release from cells into extracellular space, and the poorly understood mechanisms involved in ligand reception and pathway activation at the surface of target cells. We highlight several critical questions that remain open.


Apoptosis changes and SA-beta galactosidase expression in stress-induced premature senescence (SIPS) model of human skin fibroblasts.
Stress-induced premature senescence (SIPS) model is in vitro model of cellular aging. In this study, apoptosis was evaluated in SIPS model and in replicative senescent fibroblasts. We also compared the activity of senescence-associated beta-galactosidase (SA-beta gal) as a biomarker of cellular aging. Our results suggested that SIPS model and senescent fibroblasts might share similar mechanism of aging and apoptosis pathway.


Analytical performance of a venturi device integrated into an electrospray ionization fourier transform ion cyclotron resonance mass spectrometer for analysis of nucleic acids.
A voltage-assisted venturi device modeled after an industrial air amplifier was used to improve the ion transmission efficiency of a 16.2 kDa oligonucleotide and a 53-mer PCR product in the high-pressure region between an electrospray ionization (ESI) emitter and the sampling orifices of two Fourier transform ion cyclotron resonance mass spectrometers (FT-ICR-MS). The venturi device increased the total ion abundance of the oligonucleotide and the PCR product by more than 6-fold relative to the best achievable signal without the device. Furthermore, the average charge states of the oligonucleotide and PCR product shifted from 12.5- to 14.5- and 10.9- to 12.6-, respectively, with the addition of the venturi device. Specific to FT-ICR mass spectrometry, this increase in the charge state directly translates to an increase in theoretical mass resolving power (>10000 full width half-maximum for the results presented here at 7 T). Adduction was still observed while using the device, suggesting that it is "soft" relative to other high-pressure ion focusing methods.


Early therapeutic alliance as a predictor of treatment outcome for adolescent cannabis users in outpatient treatment.
The association of early alliance to treatment attendance and longitudinal outcomes were examined in 356 adolescents participating in a randomized clinical trial targeting cannabis use. Both patient and therapist views of alliance were examined, and outcomes were evaluated over 12 months after numerous other sources of variance were controlled. Patient-rated alliance predicted a reduction in cannabis use at three and six months and a reduction in substance-related problem behaviors at six months. Therapist-rated alliance did not predict outcomes. Neither patient nor therapist alliance ratings were associated with attendance. The findings support the important and often overlooked role that alliance can play in treating substance abusing, often delinquent, adolescents.


Unexpected gender differences in progressive supranuclear palsy reveal efficacy for davunetide in women.
Progressive supranuclear palsy (PSP) is a pure tauopathy, implicating davunetide, enhancing Tau-microtubule interaction, as an ideal drug candidate. However, pooling patient data irrespective of sex concluded no efficacy. Here, analyzing sex-dependency in a 52 week-long- PSP clinical trial (involving over 200 patients) demonstrated clear baseline differences in brain ventricular volumes, a secondary endpoint. Dramatic baseline ventricular volume-dependent/volume increase correlations were observed in 52-week-placebo-treated females (r = 0.74, P = 2.36-9), whereas davunetide-treated females (like males) revealed no such effects. Assessment of primary endpoints, by the PSP Rating Scale (PSPRS) and markedly more so by the Schwab and England Activities of Daily Living (SEADL) scale, showed significantly faster deterioration in females, starting at trial week 13 (P = 0.01, and correlating with most other endpoints by week 52). Twice daily davunetide treatments slowed female disease progression and revealed significant protection according to the SEADL scale as early as at 39 weeks (P = 0.008), as well as protection of the bulbar and limb motor domains considered by the PSPRS, including speaking and swallowing difficulties caused by brain damage, and deterioration of fine motor skills, respectably (P = 0.01), at 52 weeks. Furthermore, at 52 weeks of trial, the exploratory Geriatric Depression Scale (GDS) significantly correlated with the SEADL scale deterioration in the female placebo group and demonstrated davunetide-mediated protection of females. Female-specific davunetide-mediated protection of ventricular volume corresponded to clinical efficacy. Together with the significantly slower disease progression seen in men, the results reveal sex-based drug efficacy differences, demonstrating the neuroprotective and disease-modifying impact of davunetide treatment for female PSP patients.


Frontier Orbital Engineering of Metal-Organic Frameworks with Extended Inorganic Connectivity: Porous Alkaline-Earth Oxides.
The development of conductive metal-organic frameworks is challenging owing to poor electronic communication between metal clusters and the organic ligands that bridge them. One route to overcoming this bottleneck is to extend the inorganic dimensionality, while using the organic components to provide chemical functionality. Using density functional theory methods, we demonstrate how the properties of the alkaline-earth oxides SrO and BaO are transformed upon formation of porous solids with organic oxygen sources (acetate and trifluoroacetate). The electron affinity is significantly enhanced in the hybrid materials, while the ionization potential can be tuned over a large range with the polarity of the organic moiety. Furthermore, because of their high-vacuum fraction, these materials have dielectric properties suitable for low-κ applications.


Mitochondria-targeted antioxidant SkQ1 improves spermatogenesis in Immp2l mutant mice.
Previous studies have confirmed that spermatogenesis in homozygous Immp2l mutant male mice was normal at the age of 6 months, but was significantly abnormal at the age of 13 months. Meanwhile, oxidative stress is reported to be involved in spermatogenic impairment in old mutant mice. However, it is unclear whether antioxidant treatment is a suitable intervention for improving spermatogenesis in old mutant mice. This study sought to investigate the effect of mitochondria-targeted antioxidant SkQ1 on spermatogenesis in homozygous Immp2l mutant mice. Immp2l mutant mice were treated with the mitochondria-targeted antioxidant SkQ1 from the age of 6 weeks until 13 months. SkQ1 treatment significantly improved spermatogenesis in old Immp2 l mutant mice. Moreover, SkQ1 treatment improved the morphology of testicular seminiferous tubules, significantly reduced the apoptosis of germ cells and increased the level of GPX4 expression in old Immp2 l mutant mice. In conclusion, our data suggest that the mitochondria-targeted antioxidant SkQ1 is effective in improving spermatogenesis in Immp2 l mutant mice and might be used for the treatment of male infertility.


Food deprivation decreases responsiveness of ventromedial hypothalamic neurons to melanocortins.
The melanocortin system is involved in regulation of food intake and energy balance. Melanocyte-stimulating hormone (alpha-MSH) is an endogenous melanocortin receptor (MC-R) agonist. It acts on MC3/4 receptors to reduce appetite and to increase energy expenditure. The production of alpha-MSH is reduced during food deprivation, but MC4-R density is increased. The net effect of reduced alpha-MSH production and increased receptor level is not clear. To address this question, responses of ventromedial hypothalamic (VMH) neurons to melanotan II (MTII; a synthetic analogue of alpha-MSH) were recorded in brain slices from fed and food-deprived rats. Responses to the highest dose MTII were observed in 61% of VMH neurons from fed rats but only 33% of VMH neurons from food-deprived rats. To assess a possible mechanism by which responsiveness to melanocortins is diminished even though receptor number is augmented during fasting, we examined the effect of agouti gene-related peptide (AGRP), an endogenous MC-R antagonist that stimulates food intake. The synthesis of AGRP increases during fasting. AGRP significantly reduced VMH responsiveness to MTII. Additionally, AGRP by itself evoked neuronal responses, in contrast to synthetic MC-R antagonists. AGRP (1 nM) induced a predominant inhibitory effect on VMH neurons in food-deprived rats but not in fed rats. In the presence of AGRP, MTII induced a significant inhibition of neuronal activity in deprived rats, but not in fed rats. Inhibition of VMH neurons reduces energy expenditure and the satiety signal. These findings suggest that although food deprivation increases MC4-R density, it nevertheless reduces the effectiveness of melanocortins on VMH neurons, possibly by the involvement of AGRP.


Prediction and in vitro verification of potential CTL epitopes conserved among PRRSV-2 strains.
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the causative agent of one of the most important porcine diseases with a high impact on animal health, welfare, and production economy. PRRSV exhibits a multitude of immunoevasive strategies that, in combination with a very high mutation rate, has hampered the development of safe and broadly protective vaccines. Aiming at a vaccine inducing an effective cytotoxic T cell response, a bioinformatics approach was taken to identify conserved PRRSV-derived peptides predicted to react broadly with common swine leukocyte antigen (SLA) class I alleles. Briefly, all possible 9- and 10-mer peptides were generated from 104 complete PRRSV type 2 genomes of confirmed high quality, and peptides with high binding affinity to five common SLAs were identified combining the NetMHCpan and positional scanning combinatorial peptide libraries binding predictions. Predicted binders were prioritized according to genomic conservation and SLA coverage using the PopCover algorithm. From this, 53 peptides were acquired for further analysis. Binding affinity and stability of a subset of 101 peptide-SLA combinations were validated in vitro for 4 of the 5 SLAs. Eventually, 23% of the predicted peptide-SLA combinations showed to form complexes with a dissociation half-life ≥30 min. Additionally, combining the two prediction methods proved to be more robust across alleles than either method used alone in terms of predicted-to-observed correlations. In summary, our approach represents a finely tuned epitope prediction pipeline providing a rationally selected ensemble of peptides for future in vivo experiments with pigs expressing the included SLAs.


Acute management of the upper airway in facial burns and smoke inhalation.
Among 740 patients with acute burns who were admitted to our burn center from 1972 through, 1975, thirty-six required upper airway access within the first 24 hours after burn for oral and facial burns or smoke inhalation. Nasotracheal intubation was initially used. Twelve survived; 11 were successfully extubated and one required a tracheostomy. If the patient had not sustained major smoke inhalation, extubation was usually possible without tracheostomy when edema subsided between one and six days after the burn. It is concluded that endotracheal intubation is a satisfactory method of gaining airway control in severe oral and facial burns and in smoke inhalation. The mortality associated with orofacial burns or smoke inhalation is related to the degree of lung damage, patients' s age, and the extent of the burn; it is not related to the method of upper airway control.


Assessment of seasickness and its consequences by a method of peer evaluation.
A Seasickness Questionnaire, based on a peer-rating technique, was administered to 172 Israeli Navy sailors. Use of interrater agreement as a reliability index and two cutoff points for the partition of the sample resulted in the elimination of about one-third of the initial sampl and the formation of two subsamples-the "sick" (N equals 45) and "not sick" (N equals 73) sailors. By using differentially weighted scores for various intensities of seasickness, it was demonstrated that two normally confounded aspects of seasickness can be assessed independently: physical symptoms, which constitute the apparent severity of the sickness (A.S.S.), and level of actual performance (A.P.), which reflects the individual's attempts to cope with the sickness. A.S.S. and A.P. showed differential relationships with variables such as personality disposition of active coping, military rank, and sociometric status.


De novo mutations in GRIN1 cause extensive bilateral polymicrogyria.
Polymicrogyria is a malformation of cortical development. The aetiology of polymicrogyria remains poorly understood. Using whole-exome sequencing we found de novo heterozygous missense GRIN1 mutations in 2 of 57 parent-offspring trios with polymicrogyria. We found nine further de novo missense GRIN1 mutations in additional cortical malformation patients. Shared features in the patients were extensive bilateral polymicrogyria associated with severe developmental delay, postnatal microcephaly, cortical visual impairment and intractable epilepsy. GRIN1 encodes GluN1, the essential subunit of the N-methyl-d-aspartate receptor. The polymicrogyria-associated GRIN1 mutations tended to cluster in the S2 region (part of the ligand-binding domain of GluN1) or the adjacent M3 helix. These regions are rarely mutated in the normal population or in GRIN1 patients without polymicrogyria. Using two-electrode and whole-cell voltage-clamp analysis, we showed that the polymicrogyria-associated GRIN1 mutations significantly alter the in vitro activity of the receptor. Three of the mutations increased agonist potency while one reduced proton inhibition of the receptor. These results are striking because previous GRIN1 mutations have generally caused loss of function, and because N-methyl-d-aspartate receptor agonists have been used for many years to generate animal models of polymicrogyria. Overall, our results expand the phenotypic spectrum associated with GRIN1 mutations and highlight the important role of N-methyl-d-aspartate receptor signalling in the pathogenesis of polymicrogyria.