Extension of the short wavelength side of fluorescent proteins using hydrated chromophores, and its application.
To perform correlation analysis between different physiological parameters using fluorescent protein-based functional probes, diversification of wavelength properties of fluorescent proteins is underway. However, the shortest emission wavelength of fluorescent proteins has not been updated for more than 10 years. Here, we report the development of Sumire, a fluorescent protein emitting 414 nm violet fluorescence from a hydrated chromophore. The Sumire's fluorescence property allows for the creation of FRET probes that can be used simultaneously with CFP-YFP based FRET probes for multi-parameter analysis.


Increase in P-glycoprotein accompanied by activation of protein kinase Calpha and NF-kappaB p65 in the livers of rats with streptozotocin-induced diabetes.
It is known that protein kinase C (PKC) signal transduction is enhanced in a diabetic state, and that PKC activator phorbol esters increase the gene expression of MDR1 in human tumor cells. To clarify the expression of the liver transporters under diabetic conditions and the roles of PKCalpha and the transcription factor NF-kappaB, we investigated the expression levels of Mdr1a, Mdr1b, Mdr2, Mrp2, Bcrp, Bsep, Oct1, Oat2, and Oat3 transporters, PKCalpha, IkappaB, and NF-kappaB in the liver of rats with STZ-induced hyperglycemia. A selective increase in the gene expression of Mdr1b was detected by RT-PCR. Western blotting with C219 antibody revealed an increase in P-glycoprotein. Although the mRNA level of PKCalpha was not affected, translocation of PKCalpha to the microsomal fraction was detected. NF-kappaB p65, IkappaBalpha and IkappaBbeta mRNA levels were increased as was the level of nuclear NF-kappaB p65. From these findings, it was suggested that STZ-induced hyperglycemia caused the upregulation of Mdr1b P-gp expression through the activation of PKCalpha and NF-kappaB.


Stabilizing MXene by Hydration Chemistry in Aqueous Solution.
MXenes attract interest in diverse fields but suffer from fast structural degradation by attacking of dissolved oxygen and water molecules in aqueous solution. This drawback hinders the long-term storage, applications and understanding of the chemical nature of MXenes. Herein, we report a cost-effective and environmentally sustainable way for long-term storage of MXenes in aqueous solution by hydration chemistry of nontoxic inorganic salts. The attacking of MXene by free water and dissolved oxygen molecules is inhibited by decreasing the water activity, which simultaneously lowers the dissolved oxygen concentration, of saline solution. As a result, the storage life of MXene can be prolonged to up to 400 days at ambient conditions without loss of intrinsic surface chemistry and bulk carrier properties. Over 90 % of salt protectant can be recycled by simply evaporating the final waste liquor after fully extracting the MXene to minimize the waste discharge and processing cost. This work offers a commercializable approach with high cost-effectiveness, processing sustainability and environmental benefit for extending the shelf life of MXenes.


Rickettsia species in Dermacentor reticulatus ticks feeding on human skin and clinical manifestations of tick-borne infections after tick bite.
Dermacentor reticulatus ticks are sporadically removed from human skin and therefore the medical consequences of their feeding are neglected compared to Ixodes ricinus. We investigated the prevalence of pathogens in D. reticulatus removed from human skin and possible clinical manifestations suggestive of tick-borne diseases after a tick bite. A total of 2153 ticks were studied and of these only 34 were D. reticulatus. The mean prevalence of Rickettsia in D. reticulatus was 50.0% and R. raoultii was identified in 82.4% of infected D. reticulatus ticks. We confirmed the first case of R. aeschlimannii infection in D. reticulatus ticks. Among participants bitten by D. reticulatus, 13.3% reported reddening around the tick bite site and flu-like symptoms, including lymphadenopathy and 3.3% reported eschar on the tick site bite. All of the participants with flu-like symptoms after tick removal were bitten by ticks infected with R. raoultii. The results of this study indicate that even though D. reticulatus ticks bite humans sporadically, pathogenic Rickettsia have a remarkably high prevalence in this tick species. We can expect that the incidence of tick-borne lymphadenopathy might increase with the reported expansion of the D. reticulatus into new areas and its growing abundance in Central Europe.


Cytokine Release Syndrome and Sepsis: Analogous Clinical Syndromes with Distinct Causes and Challenges in Management.
Both cytokine release syndrome (CRS) and sepsis are clinical syndromes rather than distinct diseases and share considerable overlap. It can often be challenging to distinguish between the two, but it is important given the availability of targeted treatment options. In addition, several other clinical syndromes overlap with CRS and sepsis, further making it difficult to differentiate them. This has particularly been highlighted in the recent coronavirus disease-2019 pandemic. As we start to understand the differences in the inflammatory markers and presentations in these syndromes, hopefully we will be able to enhance treatment and improve outcomes.


Mutagenicity assessment strategy for pharmaceutical intermediates to aid limit setting for occupational exposure.
Pharmaceutical intermediates (IM) are used in the synthesis of active pharmaceutical ingredients. They are not intended for human administration, yet employees may be exposed to IM during the manufacturing process. In the context of occupational health, hazard assessment of IM is needed to identify potential intrinsic hazards which could cause unwanted adverse effects. In particular, a carcinogenic potential influences the protection strategy in the workplace. DNA reactive substances may, even if present at very low levels, lead to mutations and therefore, potentially cause cancer. The use of in silico methods to predict mutagenicity is increasingly acknowledged and implemented in the recently released ICH M7 guideline for the limitation of DNA reactive impurities. In this study we investigate the possibility to apply (quantitative) structure-activity-relationships ((Q)SARs) during hazard identification to reduce the number of Ames tests needed for a hazard assessment of IM while maintaining high standards of protection of employees. Ames test outcomes for 188 substances used in the pharmaceutical production were compared with their in silico predictions using two different (Q)SAR methodologies (knowledge based and statistical) complemented by expert knowledge. The results of the analysis showed that a negative prediction for mutagenicity provides a high confidence that the IM is not mutagenic in the Ames test with the negative predictive value of 97%. On the other hand the positive predictive value was only 57% and therefore considered too low to reliably consider positive predicted IM to be mutagenic. In order to avoid any unnecessary burden for occupational health purposes caused by falsely positive predicted IM, all positive predicted IM and those with insufficient coverage by the in silico systems are submitted to an Ames test to verify or reject the prediction. It is shown that the described in silico prediction approach ensures appropriate protection strategy of the employees. Resources for performing Ames tests which do not add additional or new information for the purpose of hazard assessment could be reduced.


Comparative Studies of the Biosynthetic Gene Clusters for Anthraquinone-Fused Enediynes Shedding Light into the Tailoring Steps of Tiancimycin Biosynthesis.
Comparative analyses of the four known anthraquinone-fused enediynes biosynthetic gene clusters identified four genes, tnmE6, tnmH, tnmL, and tnmQ, unique to the tnm gene cluster. Larger scale fermentation of both the S. sp. CB03234 wild-type and the Δ tnmH and Δ tnmL mutant strains resulted in the characterization of 20 new tiancimycin (TNM) congeners, including five enediynes. These findings enabled a proposal for the late stage of TNM biosynthesis featuring an intermediate possibly common for all anthraquinone-fused enediynes.


Vasospastic angina: A literature review of current evidence.
Vasospastic angina (VSA) is a variant form of angina pectoris, in which angina occurs at rest, with transient electrocardiogram modifications and preserved exercise capacity. VSA can be involved in many clinical scenarios, such as stable angina, sudden cardiac death, acute coronary syndrome, arrhythmia or syncope. Coronary vasospasm is a heterogeneous phenomenon that can occur in patients with or without coronary atherosclerosis, can be focal or diffuse, and can affect epicardial or microvasculature coronary arteries. This disease remains underdiagnosed, and provocative tests are rarely performed. VSA diagnosis involves three considerations: classical clinical manifestations of VSA; documentation of myocardial ischaemia during spontaneous episodes; and demonstration of coronary artery spasm. The gold standard diagnostic approach uses invasive coronary angiography to directly image coronary spasm using acetylcholine, ergonovine or methylergonovine as the provocative stimulus. Lifestyle changes, avoidance of vasospastic agents and pharmacotherapy, such as calcium channel blockers, nitrates, statins, aspirin, alpha1-adrenergic receptor antagonists, rho-kinase inhibitors or nicorandil, could be proposed to patients with VSA. This review discusses the pathophysiology, clinical spectrum and management of VSA for clinicians, as well as diagnostic criteria and the provocative tests available for use by interventional cardiologists.


Rapid Growth of Colloidal Crystal Films from the Concentrated Aqueous Ethanol Suspension.
Developing a rapid fabrication of colloidal crystal film is one of the technical issues to apply to wide and various fields. We have been investigating a drying process of colloidal aqueous ethanol (EtOH) suspension formed by electrophoretic deposition (EPD). Here, the detailed formation mechanism of the colloidal crystal films with the closest packing structure was investigated by optical microscope and spectroscopy. The growth mechanism from the colloidal suspension to the colloidal crystal film was found to consist of four stages. In the first stage, concentrated colloidal suspension changed to order structure, i.e., nonclosely packed colloidal crystal by Alder phase transition. After this crystallization, we observed Bragg's diffraction peak and structural color. In the second stage, the diffraction peak shifts toward the shorter-wavelength direction (blue shift) due to the reduction of the interparticle distance of the nonclosely packed colloidal crystal. Finally, this peak shift continued until the closely packed colloidal crystal film was formed. In the third stage, the diffraction peak kept almost a similar wavelength due to the liquid film of aqueous EtOH covering on the colloidal crystal film. In the fourth stage, the colloidal crystal film changed from wet to dry condition. The structural color changes from green to blue by the evaporation of the solvent from the interspace of the colloidal crystal film. This color change is explained by the change in the refractive index of the interparticle medium from solvent to air. One of the key findings in our process is a rapid crystal growth using concentrated colloid aqueous EtOH suspension. Drying the concentrated suspension formed a closely packed colloidal crystal film within 55 s. This process has the potential for high-speed deposition of the colloidal crystalline thin films.


Progressive Surgical Autonomy Observed in a Hand Surgery Resident Clinic Model.
Resident clinics (RCs) are intended to catalyze the achievement of educational milestones through progressively autonomous patient care. However, few studies quantify their effect on competency-based surgical education, and no previous publications focus on hand surgery RCs (HRCs). We demonstrate the achievement of progressive surgical autonomy in an HRC model. A retrospective review of all patients seen in a weekly half-day HRC from October 2010 to October 2015 was conducted. Investigators compiled data on patient demographics, provider encounters, operational statistics, operative details, and dictated surgical autonomy on an ascending 5 point scoring system. A tertiary hand surgery referral center. A total of 2295 HRC patients were evaluated during the study period in 5173 clinic visits. There was an average of 22.6 patients per clinic, including 9.0 new patients with 6.5 emergency room referrals. Totally, 825 operations were performed by 39 residents. Trainee autonomy averaged 2.1/5 (standard deviation [SD] = 1.2), 3.4/5 (SD = 1.3), 2.1/5 (SD = 1.3), 3.4/5 (SD = 1.2), 3.2/5 (SD = 1.5), 3.5/5 (SD = 1.5), 4.0/5 (SD = 1.2), 4.1/5 (SD = 1.2), in postgraduate years 1 to 8, respectively. Linear mixed model analysis demonstrated training level significantly effected operative autonomy (p = 0.0001). Continuity of care was maintained in 79.3% of cases, and patients were followed an average of 3.9 clinic encounters over 12.4 weeks. Our HRC appears to enable surgical trainees to practice supervised autonomous surgical care and provide a forum in which to observe progressive operative competency achievement during hand surgery training. Future studies comparing HRC models to non-RC models will be required to further define quality-of-care delivery within RCs.


Design and rationale of a randomized controlled trial to reduce cardiovascular disease risk for patients with bipolar disorder.
Persons with bipolar disorder (BD) experience a disproportionate burden of medical comorbidity, notably cardiovascular disease (CVD), contributing to decreased function and premature mortality. We describe the design, rationale, and baseline findings for the Self-Management Addressing Heart Risk Trial (SMAHRT), a randomized controlled effectiveness trial of an intervention (Life Goals Collaborative Care; LGCC) designed to reduce CVD risk factors and improve physical and mental health outcomes in patients with BD. Patients with BD and at least one CVD risk factor were recruited from a VA healthcare system and randomized to either LGCC or usual care (UC). LGCC participants attended four weekly, group-based self-management sessions followed by monthly individual contacts supportive of health behavior change and ongoing medical care management. In contrast, UC participants received monthly wellness newsletters. Physiological and questionnaire assessments measured changes in CVD outcomes and quality of life (QOL) over 24 months. Out of the 180 eligible patients, 134 patients were enrolled (74%) and 118 started the study protocols. At baseline (mean age=54, 17% female, 5% African American) participants had a high burden of clinical risk with nearly 70% reporting at least three CVD risk factors including, smoking (41%) and physical inactivity (57%). Mean mental and physical HRQOL scores were 1.5 SD below SF-12 population averages. SMAHRT participants experienced substantial CVD morbidity and risk factors, poor symptom control, and decreased QOL. LGCC is the first integrated intervention for BD designed to mitigate suboptimal health outcomes by combining behavioral medicine and care management strategies.


Crystal structure of (7-{[bis-(pyridin-2-ylmeth-yl)amino-κ3 N,N',N'']meth-yl}-5-chloro-quinolin-8-ol)di-bromidozinc(II).
In the title compound, [ZnBr2(C22H19ClN4O)], the ZnII atom adopts a distorted square-pyramidal coordination geometry, formed by two bromido ligands and three N atoms of the bis-(pyridin-2-ylmeth-yl)amine moiety in the penta-dentate ligand containing quinolinol. The ZnII atom is located well above the mean basal plane of the square-based pyramid. The apical position is occupied by a Br atom. The O and N atoms of the quinolinol moiety in the ligand are not coordinated to the ZnII atom. An intra-molecular O-H⋯N hydrogen bond, generating an S(5) ring motif, stabilizes the mol-ecular structure. In the crystal, the mol-ecules are linked by inter-molecular C-H⋯Br hydrogen bonds, generating ribbon structures containing alternating R 2 2(22) and R 2 2(14) rings. These ribbons are linked through an inter-molecular C-H⋯Br hydrogen bond, forming a two-dimensional network sheet.


A simple C-reactive protein measurement for the differentiation between tuberculous and malignant pleural effusion.
The aim of this study was to determine the validity of pleural fluid C-reactive protein (CRP) concentrations and/or pleural fluid to serum CRP ratio for differentiating tuberculous pleuritis (TBP) from malignant pleural effusion (MPE) in patients presenting with lymphocytic exudative pleural effusions. A cross-sectional study was conducted on 161 patients with pleural effusion who underwent diagnostic evaluation at Siriraj Hospital, Bangkok, Thailand, between April 2001 and March 2002. The complete biochemical analysis of pleural fluid, cultures of pleural fluid, and pathological examinations of pleural fluid and pleural tissue were performed. The CRP concentrations were then measured in stored sera and pleural fluid samples from patients with a lymphocytic exudative pleural effusion and with a definite diagnosis. Among the 148 patients with lymphocytic exudative pleural effusions, 55 were diagnosed with TBP, 60 with MPE, and 33 with non-specific pleuritis. Pleural fluid and serum CRP levels were significantly higher in the TBP group than in the MPE group (54.58 +/- 4.50 mg/L and 106.93 +/- 9.54 mg/L vs 12.66 +/- 3.52 mg/L and 49.66 +/- 8.84 mg/L, respectively, P < 0.001). The ratio of pleural fluid to serum CRP was significantly higher in the TBP group than in the MPE group (0.52 +/- 0.18 vs 0.30 +/- 0.16, P < 0.001). The optimum cut-off value for pleural fluid CRP level of > or =30 mg/dL had a sensitivity of 72% with 93% specificity, and the pleural fluid to serum CRP ratio cut-off value of 0.45 had a sensitivity of 60% with 89% specificity. A correlation between serum and pleural fluid CRP levels was observed in TBP patients but not in MPE patients. In patients presenting with lymphocytic exudative pleural effusion, a simple marker of raised pleural fluid CRP level may be helpful in discriminating between TBP and MPE.


Effects of health concept model-based detailed behavioral care on mood and quality of life in elderly patients with chronic heart failure.
With the intensification of social aging, the susceptibility of the elderly population to diseases has attracted increasing attention, especially chronic heart failure (CHF) that accounts for a large proportion of the elderly. To evaluate the application value of health concept model-based detailed behavioral care in elderly patients with CHF. This study recruited 116 elderly CHF patients admitted from October 2018 to October 2020 and grouped them according to the nursing care that they received. The elderly patients who underwent health concept model-based detailed behavioral care were included in a study group (SG; n = 62), and those who underwent routine detailed behavioral nursing intervention were included as a control group (CG; n = 54). Patients' negative emotions (NEs), quality of life (QoL), and nutritional status were assessed using the self-rating anxiety/ depression scale (SAS/SDS), the Minnesota Living with Heart Failure Ques-tionnaire (MLHFQ), and the Modified Quantitative Subjective Global Assessment (MQSGA) of nutrition, respectively. Differences in rehabilitation efficiency, NEs, cardiac function (CF) indexes, nutritional status, QoL, and nursing satisfaction were comparatively analyzed. A higher response rate was recorded in the SG vs the CG after intervention (P < 0.05). After care, the left ventricular ejection fraction was higher while the left ventricular end-diastolic dimension and left ventricular end systolic diameter were lower in the SG compared with the CG (P < 0.05). The post-intervention SAS and SDS scores, as well as MQSGA and MLHFQ scores, were also lower in the SG (P < 0.05). The SG was also superior to the CG in the overall nursing satisfaction rate (P < 0.05). Health concept model-based detailed behavioral care has high application value in the nursing care of elderly CHF patients, and it can not only effectively enhance rehabilitation efficiency, but also mitigate patients' NEs and improve their CF and QoL.


Preparation, characterization, antioxidant evaluation of new curcumin derivatives and effects of forming HSA-bound nanoparticles on the stability and activity.
Curcumin (CCM) is a well-known active component, which has been studied extensively in food and medicine field since it showed various activities. However, some serious issues limit its application, for example, the extremely low solubility, stability and bioavailability. In this study, 10 Curcumin derivatives were synthesized and characterized by 1H NMR, 13C NMR and HR-MS, then their antioxidant activity was evaluated. Compound 2 and curcumin were further investigated by preparing HSA-bound nanoparticles (NP-2 and NP-CCM) to surmount the difficulties mentioned above. The nanoparticles obtained were about 110 nm in size measured by Dynamic light scattering (DLS), the stability of compound 2 in NP-2 was significantly increased. Above all, NP-2 showed more efficient antioxidant and antitumor activity, which was probably attributed to the introduced isopentenyl groups in 2, it was supposed that the isopentenyl groups increased the interaction between compound 2 and HSA. Overall, NP-2 has great potential for some food and pharmaceutical applications.


Evidence for clustering of hepatitis B virus infection in families of patients with primary hepatocellular carcinoma.
Family member of 13 patients with hepatitis B surface antigen (HBsAg) positive primary hepatocellular carcinoma (PHC) were tested for the presence of hepatitis B virus-associated antigens and antibodies. Of the 122 members examined, circulating HGsAg was detected in 47 (39%), antibody to HBsAg (anti-HBs) was found in 37 (30%), and antibody to hepatitis B core antigen (anti-HBc) alone was present in 13 (11%). The relatives with the highest frequency of HBsAg positivity were the offspring of the propositus, followed by the nieces and nephews and the grandchildren. Anti-HBs and anti-HBc were detected most often in the spouses and non-blood relatives. Evidence for past and present hepatitis B virus (HBV) infection was more frequently found in the Asian family members when compared to the non-Asians. The e antigen (HBeAg) was present in 38% of the HBsAg positive individuals, including four with PHC; antibody to HBcAg (anti-HBe) was rarely detected. These results indicate that clustering of HBV infection was commonly present in family members of patients with PHC. The HBsAg positive individuals may be major contributors to the endemic pool of the virus, and may themselves be potential cases of chronic active type B hepatitis, cirrhosis, and PHC.


Periapical surgery in a Norwegian county hospital: follow-up findings of 477 teeth.
Four-hundred seventy-seven teeth treated by periapical surgery by one surgeon were followed up until a stable radiographic situation was recorded. Radiographic findings were classified into one of four groups: 1, complete healing; 2, incomplete healing (scar tissue); 3, uncertain healing; or 4, unsatisfactory healing. Evaluation was performed independently by an endodontist and the oral surgeon and cases of disagreement were subjected to joint evaluation. Difficult cases and borderline cases were judged by an oral radiologist. Complete healing was observed for 78% and incomplete healings (scar tissue) for 9% of the teeth. The size of the latter group was strongly influenced by the number of cases with large preoperative lesions. When the material was grouped as "success" (complete healing, scar tissue healing) and "failure" (uncertain healing, unsatisfactory healing), a 13% failure frequency was recorded. Twenty-eight percent of the cases treated with retrograde fills failed compared with 4% in the orthograde group.


SmartArm: Integration and validation of a versatile surgical robotic system for constrained workspaces.
With the increasing presence of surgical robots minimally invasive surgery, there is a growing necessity of a versatile surgical system for deep and narrow workspaces. We developed a versatile system for constrained workspaces called SmartArm. It has two industrial-type robotic arms with flexible tools attached to its distal tip, with a total of nine active degrees-of-freedom. The system has a control algorithm based on constrained optimization that allows the safe generation of task constraints and intuitive teleoperation. The SmartArm system is evaluated in a master-slave experiment in which a medically untrained user operates the robot to suture the dura mater membrane at the skull base of a realistic head phantom. Our results show that the user could accomplish the task proficiently, with speed and accuracy comparable to manual suturing by surgeons. Conclusions We demonstrated the integration and validation of the SmartArm.


Incorporation of ABCB1-mediated transport into a physiologically-based pharmacokinetic model of docetaxel in mice.
Docetaxel is one of the most widely used anticancer agents. While this taxane has proven to be an effective chemotherapeutic drug, noteworthy challenges exist in relation to docetaxel administration due to the considerable interindividual variability in efficacy and toxicity associated with the use of this compound, largely attributable to differences between individuals in their ability to metabolize and eliminate docetaxel. Regarding the latter, the ATP-binding cassette transporter B1 (ABCB1, PGP, MDR1) is primarily responsible for docetaxel elimination. To further understand the role of ABCB1 in the biodistribution of docetaxel in mice, we utilized physiologically-based pharmacokinetic (PBPK) modeling that included ABCB1-mediated transport in relevant tissues. Transporter function was evaluated by studying docetaxel pharmacokinetics in wild-type FVB and Mdr1a/b constitutive knockout (KO) mice and incorporating this concentration-time data into a PBPK model comprised of eight tissue compartments (plasma, brain, heart, lung, kidney, intestine, liver and slowly perfused tissues) and, in addition to ABCB1-mediated transport, included intravenous drug administration, specific binding to intracellular tubulin, intestinal and hepatic metabolism, glomerular filtration and tubular reabsorption. For all tissues in both the FVB and KO cohorts, the PBPK model simulations closely mirrored the observed data. Furthermore, both models predicted AUC values that were with 15 % of the observed AUC values, indicating that our model-simulated drug exposures accurately reflected the observed tissue exposures. Overall, our PBPK model furthers the understanding of the role of ABCB1 in the biodistribution of docetaxel. Additionally, this exemplary model structure can be applied to investigate the pharmacokinetics of other ABCB1 transporter substrates.


Development and distribution of cysts of an avirulent strain of Toxoplasma and the humoral immune response in mice.
An avirulent T. gondii strain isolated from a owl (Glaucidium brasilianum) produces randomly distributed cysts in the brain of mice which can survive to inocula as high as 1000 oocysts. Cysts appeared for the first time after 15 days of infection. Regarding to humoral immunity development due to our TCR-2 strain, detectable antibodies were found after 12 days of infection.


Counter-regulation of rejection activity against human liver grafts by donor PD-L1 and recipient PD-1 interaction.
Co-inhibitory receptor-ligand interactions fine-tune immune responses by negatively regulating T cell functions. Our aim is to examine the involvement of co-inhibitory receptor-ligand pair PD-1/PD-L1 in regulating rejection after liver transplantation (LT) in humans. PD-L1/PD-1 expression in liver allograft was determined by immunohistochemistry or flow cytometry, and the effect of blockade was studied using graft-infiltrating T cells ex vivo. Five single nucleotide polymorphisms within PD-1 and PD-L1 genes were genotyped in 528 LT recipients and 410 donors, and associations with both early (⩽6months) and late (>6months) acute rejection were analyzed using Cox proportional-hazards regression model. The effect of PD-L1 rs4143815 on PD-L1 expression was analyzed using donor hepatic leukocytes. PD-L1 was expressed by hepatocytes, cholangiocytes and along the sinusoids in post-transplant liver allografts, and PD-1 was abundantly expressed on allograft-infiltrating T cells. PD-L1 blockade enhanced allogeneic proliferative responses of graft-infiltrating T cells. In the genetic association analysis, donor PD-L1 rs4143815 (CC/CG vs. GG; HR=0.230; p=0.002) and recipient PD-1 rs11568821 (AA/AG vs. GG; HR=3.739; p=0.004) were associated with acute rejection late after LT in multivariate analysis. Recipients carrying the PD-1 rs11568821 A allele who were transplanted with liver grafts of PD-L1 rs4143815 GG homozygous donors showed the highest risk for late acute rejection. PD-L1 rs4143815 is associated with differential PD-L1 expression on donor hepatic dendritic cells upon IFN-γ stimulation. Our data suggest that interplay between donor PD-L1 and recipient PD-1 counter-regulates rejection activity against liver grafts in humans.


An atlas of genetic correlations between gestational age and common psychiatric disorders.
We aim to systematically explore the potential genetic correlations between five major psychiatric disorders and gestational ages. Genome-wide association study (GWAS) summary datasets of attention deficit/hyperactivity disorder (ADHD), autism spectrum disorder (ASD), bipolar disorder (BD), schizophrenia (SCZ) and major depressive disorder (MDD) in discovery were downloaded from the Psychiatric GWAS Consortium (PGC) website. Suggestive (Raw p < 0.05) genetic associations in the discovery phrase were further replicated in independent GWASs which downloaded from PGC, the FinnGen study or Integrative Psychiatric Research (iPSYCH) website. GWASs of gestational duration, preterm and post-term birth were derived from previous studies of infants from the Early Growth Genetics (EGG) Consortium, the iPSYCH study, and the Genomic and Proteomic Network for Preterm Birth Research (GPN). We calculated genetic correlations using linkage disequilibrium score (LDSC) regression. Mendelian randomization (MR) analyses were performed to investigate the causal effects. We identified four suggestive genetic correlations between psychiatric disorders and gestational age factors in discovery LDSC and two replicated in a confirmation LDSC: gestational duration and ADHD (rg = -0.1405, FDR p = 0.0406), post-term birth and SCZ (rg = -0.2003, FDR p = 0.0042). We also observed causal effect of post-term birth on SCZ by MR (PWeighted median = 0.037, PInverse variance weighted = 0.007). Our analysis suggested no significant evidence of horizontal pleiotropy and heterogeneity. This study showed the genetic correlation evidences between gestational age phenotypes and psychiatric disorders, providing novel clues for understanding the pathogenic factors of common psychiatric disorders. LAY SUMMARY: Whereas gestational age factors were reported to be associated with psychiatric disorders, the genetic relationship and causality remain to be revealed. The present study reported the first large-scale genetic correlations investigation of the associations between gestational age phenotypes and psychiatric disorders. Results indicate causal relationships between post-term birth and schizophrenia (SCZ), as well as suggestive genetic correlations between gestational duration and attention deficit/hyperactivity disorder (ADHD). This study provided novel clues for understanding the pathogenic factors of common psychiatric disorders.


High-resolution bioactivity profiling combined with HPLC-HRMS-SPE-NMR: α-Glucosidase inhibitors and acetylated ellagic acid rhamnosides from Myrcia palustris DC. (Myrtaceae).
Type 2 diabetes (T2D) is an endocrine metabolic disease with a worldwide prevalence of more than 8%, and an expected increase close to 50% in the next 15-20years. T2D is associated with severe and life-threatening complications like retinopathy, neuropathy, nephropathy, and cardiovascular diseases, and therefore improved drug leads or functional foods containing α-glucosidase inhibitors are needed for management of blood glucose. In this study, leaves of Myrcia palustris were investigated by high-resolution α-glucosidase inhibition profiling combined with HPLC-HRMS-SPE-NMR. This led to identification of casuarinin, myricetin 3-O-β-d-(6″-galloyl)galactopyranoside, kaempferol 3-O-β-d-galactopyranoside, myricetin, and quercetin as α-glucosidase inhibitors. In addition, four acetylated ellagic acid rhamnosides, i.e., 4-O-(2″,4″-O-diacetyl-α-l-rhamnopyranosyl)ellagic acid, 4-O-(2″,3″-O-diacetyl-α-l-rhamnopyranosyl)ellagic acid, 4-O-(3″,4″-O-diacetyl-α-l-rhamnopyranosyl)ellagic acid, and 4-O-(2″,3″,4″-O-triacetyl-α-l-rhamnopyranosyl)ellagic acid were identified.


The safety of nanocrystalline silver dressings on burns: a study of systemic silver absorption.
Wound dressings containing silver have been in widespread use for many years. However, there are few quantitative data on the systemic absorption of silver or whether there is associated clinical risk. To assess systemic silver levels when Acticoat dressings containing nanocrystalline silver were used, and to determine whether increases in such levels were associated with haematological or biochemical indicators of toxicity. A prospective, single-centre, open-label study of 30 patients with relatively small burns that required skin grafting. Serum silver levels were measured before, during and at discontinuation of the use of the Acticoat dressings, and again at 3 and 6 months following completion of treatment. The median total postoperative wound size was 12% of the total body surface area. The median time to maximum silver levels was 9 days. The median maximum serum silver level was 56.8 microg/l. The median serum level at 6 months was 0.8 microg/l. There were no haematological or biochemical indicators of toxicity associated with the silver absorption observed in this study. This study has confirmed our view that Acticoat products are safe for use on burns and they remain a standard part of treatment at our centre.


Clinical features and outcome in hospitalized heart failure in Japan (from the ATTEND Registry).
Hospitalized heart failure (HHF) is a critical issue in Japan. To improve its management and outcomes, the clinical features, in-hospital management, and outcomes should be analyzed to improve the guidelines for HHF. The acute decompensated heart failure syndromes (ATTEND) registry is the largest study of HHF in Japan. The present report covers the clinical features and in-hospital management of HHF patients. The data from 4,842 enrolled patients have demonstrated that most Japanese HHF patients are elderly, with new onset, and a history of hypertension and orthopnea on admission. During hospitalization, furosemide and carperitide were commonly used and the length of stay was extremely long (mean 30 days), with 6.4% in-hospital mortality. The findings of the present study suggest the following: (1) the focus for hypertensive elderly and diabetic patients should be on primary prevention of HHF,(2) more intensive management with noninvasive positive pressure ventilation should be performed at the urgent stage, (3) it is necessary to clarify the clinical benefit of carperitide and angiotensin-receptor blockers, because they are commonly used in Japan, and (4) it is necessary to clarify the relationship between in-hospital mortality and length of stay from the viewpoint of both outcome and cost of patient care.


Conservative management of muscle-infiltrating bladder cancer: prospective experience.
Between May 1979 and July 1983, 217 consecutive patients with documented primary bladder tumors invading muscle were evaluated to determine the fate of patients with conservatively treated muscle-infiltrating bladder cancer. The disease was re-staged by urine cytology, bimanual examination with the patient under anesthesia and transurethral biopsy or resection. Of the 217 patients 172 underwent total or partial cystectomy and 45 (21 per cent, 37 with stage T2, 7 with stage T3a and 1 with stage T4 disease) did not because re-staging showed no residual tumor (stage T0) in 20, carcinoma in situ in 17, stage T1 tumor in 4 and local stage T2 cancer in 4. The median followup was 5.1 years (range 3 to 7 years). Of the 45 patients 30 (65 per cent) are free of tumor or have required transurethral resection and intravesical therapy for recurrent tumors but cystectomy has not been necessary. Of the 15 failures 11 underwent cystectomy 9 to 30 months after re-staging (7 are alive and 4 died of disease) and 4 are alive with metastatic disease (2 with negative bladder biopsies). Re-staging in the 4 patients who died showed stage T0 disease in 2, carcinoma in situ in 1 and stage T2 tumor in 1. The over-all survival rate was 82 per cent (37 of 45) and it was 67 per cent (30 of 45) for patients with a functioning bladder. The data suggest that endoscopic re-staging may identify a subset of patients with limited muscle-infiltrating bladder tumors that can be managed conservatively without immediate cystectomy.


Primary infertility and uterine anomalies.
During a 20-year period, 19 (9.1%) of 208 patients with uterine anomalies had primary infertility. Women with unicornuate uteri had the highest (15%) incidence of primary infertility, which was found in the other groups of uterine anomalies in 7% to 13% of the patients. The cause of infertility was a nonuterine factor in 12 cases: hormonal (8), endometriosis (2), tubal (1), or male (1). The reason for infertility remained unknown or the patient conceived during investigation in five cases. Malformation of the uterus was considered the sole reason for infertility, and metroplasty was performed in two cases. During the follow-up period, 14 patients (74%) achieved pregnancy: 6 spontaneously, 3 after curettage, 2 after metroplasty, 1 after clomiphene treatment, 1 after hysterosalpingogram, and 1 after conservative endometriosis surgery. Four of five cases without pregnancy had a nonuterine factor as the cause of infertility, and in one case it may have been a uterine anomaly--a unicornuate uterus with a rudimentary horn. The results indicate that uterine anomalies are rarely the reason for infertility. Nonuterine causes of infertility must be ruled out before metroplasty is performed, as a last resort.


Occurrence and identification of Emeria species in broiler rearing under traditional system.
The present study was intended to determine the prevalence and identification of species involved causing coccidiosis in broilers rearing under traditional farming system in Mirpur, Azad Kashmir, Pakistan. For the current study, a convenient sampling technique was carried out. A total of 7814 broilers (aged 1 to 6 weeks) were submitted to the Disease Diagnostic Laboratory of Livestock and Animal husbandry Department Mirpur, Azad Kashmir. From the total screened, 750 were found positive for coccidiosis representing an overall prevalence of 9.59 %. Age-wise highest prevalence (10.88 %) recorded in the middle age birds (0 to 3 week old) were found more susceptible to infection than those aged above 3 weeks. Higher prevalence (12.49 %) of coccidiosis in broilers was observed in spring as compared with 6.60 % in summer season. In this study two main coccidiosis causing species, Emeria tenella and Emeria maxima were identified on the basis of their morphological feature and habitat (caeca and intestine), However, E. tenella was dominant compared to E. maxima. The study provides an insight to the occurrence of Emeria species which must be taken into consideration when rearing the broilers.


The effects of major abdominal surgery, enteral and parenteral nutrition on pancreatic function and morphology.
The effects of major abdominal surgery and nutritional support upon pancreatic function and morphology were studied in similar groups of patients who underwent major abdominal surgery (n = 18), received parenteral nutritional support (n = 18) or received enteral nutritional support (n = 16). The exocrine function of the pancreas was measured by means of an oral pancreatic function test (using 1 g of N-benzoyl-L-tyrosyl para-aminobenzoic acid) with measurement of serum para-aminobenzoic acid at 3 h and the Pancreatic Excretion Index. Pancreatic morphology was assessed by real time ultrasound and a pancreatic size index was calculated (maximum diameter of head x body). Serial measurements of function and morphology were carried out in each patient at entry into the study and at 7 and 14 days after operation or start of nutritional support. Serum PABA levels were similar in the 3 groups at the start of the study. Although the levels remained unchanged in the enteral and parenteral groups, a significant and progressive decrease was observed in the surgical group (serum PABA = 27.5 nmol/l (24.0-30.6) before operation, 10.5 nmol/l (5.5-13.4) 14 days after operation, P < 0.01). No significant changes in pancreatic size were seen. Parenteral and enteral nutrition preserves pancreatic exocrine function. A dramatic reduction in pancreatic exocrine function is seen after major abdominal surgery.


Determination, residue analysis, risk assessment and processing factors of tebufenozide in okra fruits under field conditions.
Ensuring the yield, quality, and profitability of okra by preventing and controlling pests with the application of insecticides has increased in the last decade. Some insecticide residues might remain in edible parts of okra (fruits) and lead to several potential human health problems. Therefore, research on the residue behaviour, risk assessment and removal approach of insecticides on okra fruits is important for food safety, together with the proper application and residual elimination of insecticides in okra. A simple liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was established and validated for determining the tebufenozide residues in okra fruits. The recoveries of tebufenozide in okra fruits were >72% with relative standard deviations of 0.6 to 6.1%. The dissipation rates of tebufenozide were different in okra fruits cultivated under open land and glasshouse field conditions because of the discriminating humidity and temperature conditions. The dietary intake of the tebufenozide residues from okra fruit consumption for Chinese consumers was fairly low, with approximately no potential health risk. The processing factor values of washing, blanching, washing + blanching and soaking were all less than one, which indicated that these processes could effectively reduce the residual hydrazide in the okra fruit. The developed method for analysing tebufenozide in okra fruits was applicable for field studies on this insecticide. The potential health risk of tebufenozide in okra fruits could be negligible to the health of different age groups of Chinese consumers. The soaking process effectively removed tebufenozide residues from okra fruits. The obtained data will help Chinese governments establish a maximum residue limit of tebufenozide in okra and provide data for the risk assessment and removal of tebufenozide in other crops. © 2019 Society of Chemical Industry.


Successful outcome after nonmyeloablative allogeneic hematopoietic stem cell transplantation in patients with renal dysfunction.
Nonmyeloablative allogeneic hematopoietic stem cell transplantation (HSCT) is a transplantation approach that enables patients with comorbid conditions to undergo allogeneic HSCT. We investigated the outcome of patients with reduced renal function as a single comorbidity before HSCT. Thirteen patients with a glomerular filtration rate (GFR) of <60 mL/min/1.73 m2 were matched on sex, age, and type of transplant to 26 controls with normal renal function. All patients received a nonmyeloablative HSCT with fludarabine and/or total body irradiation conditioning (TBI). Graft-versus-host disease (GVHD) prophylaxis consisted of mycophenolate mofetil and cyclosporine. Data on renal function, cyclosporine dose, cyclosporine trough levels, hypertension, and GVHD were collected. Of the 13 patients with impaired renal function, 7 patients (54%) improved or stabilized to a GFR>or=60 mL/min/1.73 m2 at last follow-up. Four patients (31%) developed chronic kidney disease stage 3 (GFR <60 mL/min/1.73 m2) compared to 3 patients (12%) in the control group (P=.039). There was no difference in survival between cases and controls. Furthermore, there were no differences in complications after HSCT, and cyclosporine dose and trough levels were similar between cases and controls. Nonmyeloablative HSCT can be safely offered to patients with mildly reduced renal function. Cyclosporine can be administered at the same dose as patients without renal dysfunction, as long as cyclosporine trough levels and creatinine are monitored and dose adjustments are made if necessary.


Determining the clinical significance of the Chest Wall Injury Society taxonomy for multiple rib fractures.
The Chest Wall Injury Society (CWIS) proposals for standardized nomenclature for multiple rib fracture (MRF) classifications were derived by international expert Delphi consensus. This study aimed to validate the CWIS taxonomy using a single-instituion clinical database. Computed tomography (CT) scans, of 539 consecutive patients with MRFs admitted to a regional major trauma center over a 33-month period, were reviewed (blinded for clinical outcomes). Every rib fracture in every patient was assessed according to each of the CWIS criteria (the degree of displacement, characterization of the fracture line, location of each fracture, and the relationship to neighboring fractures). The clinical significance of the proposed CWIS definitions were determined from independently coded, routinely collected Hospital Episodes Statistics data. The radiologic aspects of 3,944 individual rib fractures were assessed. Indicators of injury severity (severe displacement greater series length, and flail segment) were positively associated with other fractures (p < 0.001), hemopneumothorax (p < 0.001), pulmonary complications (p = 0.002), adverse outcomes (p = 0.006), mechanical ventilation (p < 0.001) and prolonged hospital and intensive therapy unit length of stay (p = 0.006, p = 0.007 respectively). Four of the CWIS-proposed definitions were correlated with pulmonary complications and adverse outcomes: the categories of displacement, the definition of individual fracture characterization, the presence of a flail segment. Two definitions for which there was CWIS consensus were not correlated with clinical outcomes: the definition of a series to describe associated fractures on neighboring ribs, the inclusion of a paravertebral sector for fracture localization. The CWIS rib fracture taxonomy demonstrates clinical relevance. There were associations between the severity of category groups within three of the proposed definitions, based on the clinical outcomes observed. Clinical outcome assessment proved inconclusive for four agreed definitions. Comprehensive, multiinstitutional data collection would be required to provide validation for all the CWIS-proposed definitions. Level IV.


Macrophage-derived reactive oxygen species protects against autoimmune priming with a defined polymeric adjuvant.
Understanding the nature of adjuvants and the immune priming events in autoimmune diseases, such as rheumatoid arthritis, is a key challenge to identify their aetiology. Adjuvants are, however, complex structures with inflammatory and immune priming properties. Synthetic polymers provide a possibility to separate these functions and allow studies of the priming mechanisms in vivo. A well-balanced polymer, poly-N-isopropyl acrylamide (PNiPAAm) mixed with collagen type II (CII) induced relatively stronger autoimmunity and arthritis compared with more hydrophilic (polyacrylamide) or hydrophobic (poly-N-isopropylacrylamide-co-poly-N-tertbutylacrylamide and poly-N-tertbutylacrylamide) polymers. Clearly, all the synthesized polymers except the more hydrophobic poly-N-tertbutylacrylamide induced arthritis, especially in Ncf1-deficient mice, which are deficient in reactive oxygen species (ROS) production. We identified macrophages as the major infiltrating cells present at PNiPAAm-CII injection sites and demonstrate that ROS produced by the macrophages attenuated the immune response and the development of arthritis. Our results reveal that thermo-responsive polymers with high immune priming capacity could trigger an autoimmune response to CII and the subsequent arthritis development, in particular in the absence of NOX2 derived ROS. Importantly, ROS from macrophages protected against the autoimmune priming, demonstrating a critical regulatory role of macrophages in immune priming events.


American Society for Pain Management Nursing position statement: pain management at the end of life.
Pain at the end of life continues to be of great concern as it may be unrecognized or untreated. While nurses have an ethical obligation to reduce suffering at the end of life, barriers remain regarding appropriate and adequate pain management at the end of life. This position statement from the American Society for Pain Management Nursing contains recommendations for nurses, prescribers, and institutions that would improve pain management for this vulnerable population.


Velocity constancy and models for wide-field visual motion detection in insects.
The tangential neurons in the lobula plate region of the flies are known to respond to visual motion across broad receptive fields in visual space. When intracellular recordings are made from tangential neurons while the intact animal is stimulated visually with moving natural imagery,we find that neural response depends upon speed of motion but is nearly invariant with respect to variations in natural scenery. We refer to this invariance as velocity constancy. It is remarkable because natural scenes, in spite of similarities in spatial structure, vary considerably in contrast, and contrast dependence is a feature of neurons in the early visual pathway as well as of most models for the elementary operations of visual motion detection. Thus, we expect that operations must be present in the processing pathway that reduce contrast dependence in order to approximate velocity constancy. We consider models for such operations, including spatial filtering, motion adaptation, saturating nonlinearities, and nonlinear spatial integration by the tangential neurons themselves, and evaluate their effects in simulations of a tangential neuron and precursor processing in response to animated natural imagery. We conclude that all such features reduce interscene variance in response, but that the model system does not approach velocity constancy as closely as the biological tangential cell.


Complements in diabetes mellitus: activation of complement system evidenced by C3d elevation in IDDM.
To characterize insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM) in terms of the complement system, some components of the system as well as the related substances and indices were studied. CH50, C3, C4 and C3bINA significantly increased in both IDDM and NIDDM compared with non-diabetic healthy controls. ACH50 was also elevated in NIDDM, whereas it was similar in IDDM and controls. Besides, the serum concentration of C3d, a breakdown product of C3, was higher in IDDM than in NIDDM and healthy controls, but that in NIDDM did not differ significantly from the control. B1Hg1 was not different among IDDM, NIDDM and non-diabetic controls. These observations suggested that there is a high level of complements in both types of diabetes mellitus, but the complement activation seems to be much enhanced in IDDM compared with NIDDM.


Shared medical appointments for bariatric surgery follow-up: a patient satisfaction questionnaire.
Shared medical appointments (SMAs) are group clinics where practitioners see several patients, with common health needs, at once. There is a great financial strain on the National Health Service (NHS) to provide bariatric surgery. The aim of this study was to review patient satisfaction with the SMA that is the default means of following up patients after bariatric surgery at one particular NHS trust. A patient-validated questionnaire was designed and handed out at the end of the SMAs. Patients who attended an SMA earlier in 2011 were also retrospectively sent questionnaires via post. A total of 47 patients completed the questionnaire from seven different SMAs covering the period from January to July 2011. All patients underwent laparoscopic adjustable gastric banding. After attending an SMA, patients gave an overall mean satisfaction rating of 4.13 ± 0.163 (on a scale of 1 to 5, 1 = very poor and 5 = excellent) which represented an increase (p < 0.01) compared to preconceptions before the clinic (3.59 ± 0.175). A cost analysis estimated a yearly saving of £4,617 or 65.1% made by the SMAs compared to 1:1 appointments. The bariatric surgery SMA demonstrates high levels of patient satisfaction and is cost-effective.


New approaches to respiratory infections in children. Bronchiolitis and croup.
Croup is a disease that is commonly seen in children younger than the age of 6 years. The cause is viral, with parainfluenza viruses and RSV being the two most common pathogens. Treatment consists primarily of supportive care, and parents usually have tried humidification and cool air exposure before the child presents to the ED. Children with moderate to severe croup are usually seen in the ED. The use of steroids in an oral preparation results in a clinical improvement of outpatients with mild to moderate croup and reduces the need for hospitalization. The dosage range for oral dexamethasone is 0.15 mg/kg to 0.6 mg/kg. Nebulized budesonide may also be used. Racemic or L-epinephrine, both of which are equally effective, can be used for symptomatic treatment in severe croup. After administration of racemic or L-epinephrine, hospitalization is not automatic and patients can be discharged safely from the ED after a 3-hour of observation period. There should be no respiratory distress, and the patient should have access to follow-up and emergency care if needed.


Infective endocarditis and diabetes mellitus: Results from a single-center study from 1994 to 2017.
To evaluate the prognostic impact of diabetes mellitus (DM) in patients with Infective Endocarditis (IE). 375 patients with diagnosis of IE referred to our Hospital between 1994-2017 were retrospectively included; diabetes was reported in 129 (34.4%). Diabetic patients were older than non-diabetic (66±1 vs. 57±2 years, p<0.001) and showed a higher prevalence of comorbidities such as hypertension (75 vs. 54%, p<0.001), coronary artery disease (30 vs. 12%, p<0.001) and history of heart failure (HF; 24 vs. 13%, p = 0.021). Echocardiography showed a higher incidence of paravalvular complications (82 vs. 64%, p<0.001) and a lower left ventricular ejection fraction (LVEF; 52±11 vs. 55±10%, p = 0.001) in diabetic than in non-diabetic patients. In-hospital mortality was higher in diabetic patients (83 vs. 74%; p = 0.030). At logistic regression, history of HF (OR = 3.1, 95%CI: 1.87-5.29, p<0.001) resulted an independent predictor of in-hospital death. At long-term follow-up [median 24(7-84) months], the Kaplan-Meier analysis showed a significantly lower survival free from all-cause death in the group with diabetes (Log-rank<0.001). At the propensity score adjusted Cox multivariable analysis, DM (HR = 1.76, 95%CI: 1.18-2.6, p = 0.005), age (HR = 1.03, 95%CI: 1.02-1.05, p<0.001), intravenous drug users (HR = 5.42, 95%CI: 2.55-11.51, p<0.001) and low LVEF (HR = 0.98, 95%CI: 0.96-0.99, p = 0.013) were independently associated to a higher mortality. In patients with IE, DM is associated to a higher prevalence of anatomic complications and a more impaired LVEF. Diabetic patients show a significantly lower survival both in hospital and during follow-up compared to the non-diabetic ones.


Identification of a putative collagen-binding protein from chicken skeletal muscle as glycogen phosphorylase.
We have purified and generated antisera to a 95 kDa skeletal muscle protein that constitutes the largest mass fraction of gelatin-agarose binding proteins in skeletal muscle. Preliminary results indicated that this 95 kDa chicken skeletal muscle protein bound strongly to gelatin-agarose and type IV collagen-agarose, suggesting a possible function in muscle cell adhesion to collagen. However, N-terminal sequencing of proteolytic fragments of the 95 kDa protein indicates that it is the chicken skeletal muscle form of glycogen phosphorylase, the binding of which to gelatin-agarose is unlikely to be biologically relevant. Further characterization showed that the skeletal muscle form of glycogen phosphorylase is immunologically distinct from the liver and brain forms in the chicken, and suggests that, unlike mammalian skeletal muscle, chicken skeletal muscle may have two phosphorylase isoforms. Furthermore, immunolocalization data and solubility characteristics of glycogen phosphorylase in muscle extraction experiments suggest the enzyme may interact strongly with an unidentified component of the muscle cytoskeleton. Thus, this study yields a novel purification technique for skeletal muscle glycogen phosphorylase, provides new information on the distribution and isoforms of glycogen phosphorylase, and provides a caveat for using gelatin affinity chromatography as a primary step in purifying collagen-binding proteins from skeletal muscle.


Exploring the role of milk yield in the first week of lactation on the association between hyperketonemia and reproductive performance in dairy cattle.
The objective of this retrospective cohort study was to investigate whether the association of hyperketonemia (HYK) with reproductive performance differs based on milk production during the first week of lactation (wk 1). Data between calving and 150 d in milk (DIM) from 2,091 Holstein dairy cows from 5 commercial dairy herds in Minnesota were collected. The concentration of β-hydroxybutyrate (BHB) was measured twice between 3 and 10 DIM in whole blood to diagnose HYK (defined as BHB ≥1.2 mmol/L). The average wk 1 milk yield was classified into 3 levels (low = lower 25th percentile; mid = 25th to 75th percentile; high = upper 75th percentile) according to parity and herd. Log-binomial regression, Cox-proportional hazard regression, and linear regression models were used to investigate the association of HYK with pregnancy to first insemination, pregnancy by 150 DIM, and calving-to-conception interval (CCI) between groups, respectively. To describe the differences in these estimates between each wk 1 milk yield level, an interaction term of HYK and stratified levels of wk 1 milk yield was added. The incidence of HYK between 3 to 10 DIM was 14.2% (4.6% in primiparous cows and 19.4% in multiparous cows). In the study population, the effect of HYK on reproduction was greater in cows with low milk yield. Among cows with low milk yield levels, HYK was associated with lower risk of pregnancy to first insemination [risk ratio = 0.62; 95% confidence interval (CI): 0.39-0.98], lower risk of pregnancy up to 150 DIM (hazard ratio = 0.70; 95% CI: 0.51-0.96), and extended days of CCI (mean difference = 13.0; 95% CI: 5.1-20.9). In contrast, among cows with mid and high wk 1 milk yield levels, hyperketonemic (HYK+) cows had a similar reproductive performance to non-HYK (HYK-) cows. Our results suggest that early-lactation milk yield plays a role in the association of hyperketonemia with reproductive performance. These results merit further investigation to determine the role that early milk yield has in the association of hyperketonemia with health and productivity outcomes.


Protective effects of Salvianolic acid B on Schwann cells apoptosis induced by high glucose.
Diabetic peripheral neuropathy (DPN) is one of the most common and debilitating microvascular complications of diabetes, and there is no effective therapy for the prevention or treatment of DPN. Oxidative stress triggers several pathways of injury and may be the unifying factor of hyperglycemia. The aim of this study was to investigate protective effect of Salvianolic acid B (Sal B) on the high glucose (HG)-induced oxidative stress-induced mitochondrial pathway activation and Schwann cells (SCs) apoptosis in vitro. We found that Sal B inhibited the HG-induced oxidative stress by reducing ROS and 8-hydroxy-2-deoxy Guanosine (8-OHdG) production, and mitochondrial depolarization and apoptosis in SCs in a dose-dependent manner. Furthermore, Sal B down-regulated the HG-mediated Bax expression and AIF nuclear translocation and the release of cytochrome c, but up-regulated the HG-induced BcL-2 expression in SCs. In addition, Sal B attenuated the HG-induced activation of caspase 3 and 9 and minimized the cleavage of PARP in SCs. Our results indicated that Sal B antagonized the HG-induced oxidative stress, activation of the mitochondrial pathway and apoptosis in SCs.


Inhibition of protein tyrosine phosphatase activity in HER14 cells by melittin and Ca2+ ionophore A23187.
We investigated the effect of melittin and Ca2+ ionophore A23187 on protein tyrosine phosphatase activity in HER14 cells (NIH-3T3 cells transfected with human epidermal growth factor 'EGF' receptor). Cell fractions were used to measure protein tyrosine phosphatase activity in vitro using 32P-labeled poly(Glu/Tyr) (4:1) peptide as a substrate. Treatment of HER14 cells with melittin or with A23187, inhibited protein tyrosine phosphatase activity in the cell sonicate and homogenate, as well as in cytosolic and particulate fractions of these cells. The inhibitory effect of both drugs was prevented by preincubating cells with EGTA (ethyleneglycolbis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid). The cyclooxygenase inhibitor indomethacin enhanced the inhibitory effect of A23187, but not that of melittin. Addition of arachidonic acid to the cells partially prevented the inhibition of protein tyrosine phosphatase activity by melittin or A23187. Preexposure of cells to EGF enhanced the inhibitory effect of melittin--but not that of A23187. Addition of CaCl2, or MgCl2 to the cell homogenate inhibited protein tyrosine phosphatase activity. These results show that protein tyrosine phosphatase activity in HER14 cells is inhibited by melittin and Ca2+ ionophore A23187 through a Ca(2+)-dependent mechanism, and is regulated by arachidonic acid metabolism and EGF receptor activation.


Morphogenesis of the truncus arteriosus of the chick embryo heart: movements of autoradiographic tattoos during septation.
In order to trace tissue movements during septation of the embryonic truncus arteriosus into aortic and pulmonary cardiac outlets, the cephalic margin of the developing tubular heart of chick embryos was tattooed at Hamilton-Hamburger Stages 20-22 using diffusion micropipettes filled with 0.5% agarose and radioactive macromolecular precursors (tritiated thymidine, uridine, and leucine). Following further incubation for 2, 48, or 96 hours, the locations of such tattoos were determined by autoradiography of sectioned tissue and computer reconstruction of the developing outflow tract. Two hours after tattooing, radiolabeled cells were clustered at the right distal margin of the myocardial tube, as intended. Two days later, during septation of the outflow tract into the two arterial streams, label was concentrated along the posterior margin of the myocardium, between the developing aortic and pulmonary valve anlagen to the embryo's right and left, respectively. Four days following tattooing, as truncal septation neared completion, remaining label was found primarily to the left of the aortic valve ring posterior to the pulmonary outlet. The movements of thymidine tattoos during septation were demonstrated in a series of 31 embryos, 14 fixed at 2 hours, 12 at 2 days, and 5 at 4 days following tattooing; similar results were seen in uridine and leucine labeled hearts. The motion of such tattoos in the developing chick heart suggests that the left side of the definitive semilunar valve ring derives from the right distal margin of the primitive tubular heart and that normal morphogenesis of the great arterial streams involves both retraction and rotation of the embryonic truncus arteriosus.


Hemorrhage risk and clinical features of multiple intracranial arteriovenous malformations.
The aim of this report is to examine clinical characteristics, treatment strategies, and annual hemorrhage incidence rate for patients with multiple arteriovenous malformations (MAVM). The PubMed and EMBASE databases and the arteriovenous malformations (AVM) database at The Johns Hopkins Hospital were searched to identify patients with MAVM. Data related to demographics, clinical features, management, and treatment outcomes were analyzed with descriptive statistics. Thirty-eight patients met the inclusion criteria. The annual hemorrhage incidence rate was 6.7%. Surgical intervention remained the most common single-modality treatment from 1949-2011. Between 1990 and 2011, multiple-modality treatment strategies (36% of cases) were employed more frequently. The most common presenting features were neurological deficit (74%) and hemorrhage (63%). In patients undergoing staged treatment of MAVM, hemorrhage of an untreated nidus (n=5), visualization of a new nidus (n=9), and disappearance of an untreated nidus (n=2) were observed. Limitations of this study include small sample size and reporting bias. The annual hemorrhage incidence rate for MAVM patients was approximately two- to three-fold greater than the reported annual hemorrhage rates for solitary AVM. Combining different treatment modalities has become the most common management strategy. The potential instability of remaining nidi with staged or incomplete treatment necessitates close follow-up in these cases.


Towards mainstream partial nitritation/anammox in four seasons: Feasibility of bioaugmentation with stored summer sludge for winter anammox assistance.
The strong effect of low temperatures on anammox challenges its mainstream application over the winter in temperate climates. Winter bioaugmentation with stored summer surplus sludge is a potential solution to guarantee sufficient nitrogen removal in winter. Firstly, the systems for which nitrogen removal deteriorated by the temperature decrease (25 °C → 20 °C) could be fully restored bioaugmenting with granules resp. flocs stored for 6 months at 118 resp. 220% of the initial biomass levels. Secondly, the reactivation of these stored sludges was tested in lower temperature systems (15.3 ± 0.4/10.4 ± 0.4 °C). Compared to the activity before storage, between 56% and 41% of the activity of granules was restored within one month, and 41%-32% for flocs. Additionally, 85-87% of granules and 50-53% of flocs were retained in the systems. After reactivation (15.3 ± 0.4/10.4 ± 0.4 °C), a more specialized community was formed (diversity decreased) with Candidatus Brocadia still dominant in terms of relative abundance. Capital and operating expenditures (CAPEX, OPEX) were negligible, representing only 0.19-0.36% of sewage treatment costs.


Classic and novel stem cell niches in brain homeostasis and repair.
Neural stem cells (NSCs) critical for the continued production of new neurons and glia are sequestered in distinct areas of the brain called stem cell niches. Until recently, only two forebrain sites, the subventricular zone (SVZ) of the anterolateral ventricle and the subgranular zone (SGZ) of the hippocampus, have been recognized adult stem cell niches (Alvarez-Buylla and Lim, 2004; Doetsch et al., 1999a, 1999b; Doetsch, 2003a, 2003b; Lie et al., 2004; Ming and Song, 2005). Nonetheless, the last decade has been witness to a growing literature suggesting that in fact the adult brain contains stem cell niches along the entire extent of the ventricular system. These niches are capable of widespread neurogenesis and gliogenesis, particularly after injury (Barnabé-Heider et al., 2010; Carlén et al., 2009; Decimo et al., 2012; Lin et al., 2015; Lindvall and Kokaia, 2008; Robins et al., 2013) or other inductive stimuli (Bennett et al., 2009; Cunningham et al., 2012; Decimo et al., 2011; Kokoeva et al., 2007, 2005; Lee et al., 2012a, 2012b; Migaud et al., 2010; Pencea et al., 2001b; Sanin et al., 2013; Suh et al., 2007; Sundholm-Peters et al., 2004; Xu et al., 2005; Zhang et al., 2007). This review focuses on the role of these novel and classic brain niches in maintaining adult neurogenesis and gliogenesis in response to normal physiological and injury-related pathological cues. This article is part of a Special Issue entitled SI: Neuroprotection.


Predictive and prognostic role of tumour-infiltrating lymphocytes in breast cancer patients with different molecular subtypes: a meta-analysis.
Whether tumour-infiltrating lymphocytes (TILs) play different roles in different molecular subtypes of breast cancer remains unknown. Additionally, their prognostic and predictive value in different molecular subtypes of breast cancer is still controversial. The aim of our meta-analysis was to assess the prognostic and predictive value of TILs in different molecular subtypes of breast cancer by summarizing all relevant studies performing multivariate analysis. PubMed, Embase, EBSCO, ScienceDirect, the Cochrane Database and Web of Science were comprehensively searched (until March 2020). Hazard ratios (HRs), odds ratios (ORs) and their 95% confidence intervals (CIs) were used as effect measures to perform our meta-analysis. A random effect model was used. Stata software, version 15 (2017) (StataCorp, College Station, TX, USA) was used to perform the statistical analysis. Thirty-three studies including 18,170 eligible breast cancer patients were analysed. The meta-analysis showed that high TIL expression was significantly associated with increased pathological complete response (pCR) rates after neoadjuvant chemotherapy in patients with the HER2-enriched molecular subtype (OR = 1.137, 95% CI [1.061 ~ 1.218], p < 0.001) and triple-negative breast cancer (TNBC) subtype (OR = 1.120, 95% CI [1.061 ~ 1.182], p < 0.001). However, high TIL expression was not significantly associated with high pCR rates after neoadjuvant chemotherapy in patients with the luminal molecular subtype of breast cancer (OR = 1.154, 95% CI [0.789 ~ 1.690], p = 0.460). We carried out a meta-analysis on the HRs of overall survival (OS) and disease-free survival (DFS) to assess the prognostic value of TILs in breast cancer with different molecular subtypes more deeply. Our meta-analysis confirmed that high TILs were associated with significantly improved DFS in patients with the HER2-enriched molecular subtype [HR = 0.940, 95% CI (0.903 ~ 0.979), p = 0.003] and TNBC molecular subtype [HR = 0.907, 95% CI (0.862 ~ 0.954), p < 0.001]. However, high TILs were not associated with significantly better DFS in patients with the luminal molecular subtype of breast cancer [HR = 0.998, 95% CI (0.977 ~ 1.019), p = 0.840]. Furthermore, the results confirmed that high TILs were significantly related to better OS in patients with the HER2-enriched molecular subtype [HR = 0.910, 95% CI (0.866 ~ 0.957), p < 0.001] and TNBC molecular subtype [HR = 0.869, 95% CI (0.836 ~ 0.904), p < 0.001]. Conversely, the summarized results indicated that high TILs were significantly associated with poor OS in patients with the luminal molecular subtype of breast cancer [HR = 1.077, 95% CI (1.016 ~ 1.141), p = 0.012]. Our meta-analysis confirms that high TILs are associated with favourable survival and predicts pCR in breast cancer patients with the TNBC and HER2-enriched molecular subtypes.


Studies on platelet function during different modes of administration of PgF2alpha in obstetrics and gynecology.
Platelet function has been studied during intravenous, intraamniotic, and extraamniotic administration of prostaglandin F2alpha (PgF2alpha) for termination of missed abortion and missed labor, for therapeutic abortion, and for induction of term labor. The controls received oxytocin i.v. (missed labor and term labor). Our investigations have shown that there was a normalization of the increased spontaneous platelet aggregation and a significant reduction of ADP- and collagen-induced platelet aggregation in the groups given PgF2alpha i.v. The desaggregation in these groups was increased. The other groups given PgF2alpha showed no significant changes in platelet function. Inducing labor by oxytocin we found a tendency to increased platelet aggregation and decreased desaggregation. The clinical importance of these findings and the consequences for hemostasis are discussed.


Protocol for laser-induced chronic ocular hypertension and intracameral injection in nonhuman primates.
Laser-induced hypertension in nonhuman primates is used to mimic human glaucoma, the leading cause of irreversible blindness. In this protocol, we detail steps for laser-induced ocular hypertension in nonhuman primates by laser photocoagulation of the trabecular meshwork and subsequent intracameral injection. We further describe recording and evaluation of intraocular pressure changes and peripapillary retinal nerve fiber layer thickness. This protocol can assist researchers improve the success rate and repeatability of the procedure and reduce the number of nonhuman primates needed. For complete details on the use and execution of this protocol, please refer to Sun et al. (2022).


Systemic absorption of bacitracin after peritoneal lavage.
Nine patients underwent intraoperative peritoneal lavage with a solution containing 50,000 units of bacitracin in 200 ml of 0.9 per cent sodium chloride. The solution was removed by suction at 2 or 5 minutes, and venous blood samples were obtained at 15 minute intervals for 2 hours after lavage. Concentrations of bacitracin in serum were determined by agar diffusion assay. In the majority of patients, peak concentrations of bacitracin in serum occurred by 15 minutes post lavage. The mean peak serum concentration was 3.8 units/ml, which exceeds peak concentrations after intramuscular injection of 50,000 units. Due to potential nephrotoxicity, bacitracin should be used cautiously in peritoneal lavage solutions and should be abandoned in patients who have renal impairment in whom prolonged elevated serum concentrations could develop.


A randomized controlled trial to compare steroid with cyclosporine for the topical treatment of oral lichen planus.
To compare the effectiveness of cyclosporine solution versus triamcinolone acetonide in orabase in the treatment of oral lichen planus (OLP) in reducing signs and symptoms. One hundred thirty-nine biopsy-proven OLP patients were randomly assigned to cyclosporine (68) or steroid (71) applied onto the target lesion and affected areas. Assessments were at weeks 0, 2, 4, 8 by clinical scoring and grid measurement of the target lesion (reticulation, erythema, ulceration). Patients ranked severity of pain and burning sensation using visual analog scales. Although clinical response, pain, burning sensation, area of reticulation, erythema, and ulceration at week 4 were all worse in patients receiving cyclosporine than in those receiving steroid, the differences were not statistically significant. Large patient-to-patient variability was evident over the observation period, with little evidence of marked changes in levels over time in both treatment groups. Topical cyclosporine appears no more effective than steroid in the treatment of oral lichen planus.


In Vitro and In Vivo Approaches to Study Kinetochore-Microtubule Attachments During Mitosis.
The separation of duplicated chromosomes during mitosis is a pivotal step in the process of cellular division. Therefore, the orchestrated events that take place to ensure proper attachment and stabilization of kMTs are keen areas of interest in the mitosis field. Here we describe the methods used to study kMT attachments via in vitro biochemical methods and in vivo cell biological approaches.


Effect of inoculated azotobacteria and Phanerochaete chrysosporium on the composting of olive pomace: Microbial community dynamics and phenols evolution.
The effect of inoculated azotobacteria and basidiomycetes white-rot fungi on the population dynamics of bacteria and eumycetes during the co-composting of olive mill pomace and wheat straw was evaluated by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) analysis combined with sequencing of rRNA gene amplicons from selected DGGE bands. The evolution of pH, temperature, phytotoxicity and water-soluble phenol content during co-composting was also monitored. In general, a similar evolution of microbial biodiversity was seen in both the inoculated and uninoculated (control) piles, which was in keeping with a similar evolution of phytotoxicity and water-soluble phenol content. Overall, under the conditions applied, data suggest a marginal influence of the inoculated starters on the physical, chemical and microbiological properties of compost piles, with the resident microbiota playing a major role.


Socio-demographic factors related to parent engagement in the NICU and the impact of the SENSE program.
Early parent engagement in the neonatal intensive care unit (NICU) is important for both parent and infant mental health and for improving developmental outcomes. It remains unclear how different programs, such as the Supporting and Enhancing NICU Sensory Experiences (SENSE) program, may empower parents from various socio-demographic groups to engage in the NICU. An improved understanding could aid in individualizing interventions for those at the highest risk for health disparities. This exploratory study, which was part of a larger study, sought to explore 1) socio-demographic factors related to parent presence and engagement in the NICU and 2) if the SENSE program related to increased parent presence and engagement among different socio-demographic groups. Seventy parent-infant dyads (born ≤ 32 weeks gestation) were randomized to SENSE programming (parent education and age-appropriate, positive sensory interventions for parents to conduct with their infants every day of hospitalization) or standard care after admission to the NICU. The amount of parent presence and participation in sensory activities was tracked using bedside logs, nursing records, and research team documentation. Being married (p = 0.048; p = 0.01), having private insurance (p < 0.001; p = 0.01), and having fewer children (p = 0.004; p = 0.03) related to more parent presence and engagement respectively. Parents who were Black had less presence and engagement in the NICU (p = 0.04; p = 0.02). Participation in the SENSE program was related to more parent presence and engagement among younger mothers (p = 0.002; p ≤0.001) and among parents living farther distances from the hospital (p < 0.001; p = 0.004). Programming, such as the SENSE program, can improve parent engagement in the NICU among high-risk groups.


Injured astrocytes are repaired by Synaptotagmin XI-regulated lysosome exocytosis.
Astrocytes are known to facilitate repair following brain injury; however, little is known about how injured astrocytes repair themselves. Repair of cell membrane injury requires Ca(2+)-triggered vesicle exocytosis. In astrocytes, lysosomes are the main Ca(2+)-regulated exocytic vesicles. Here we show that astrocyte cell membrane injury results in a large and rapid calcium increase. This triggers robust lysosome exocytosis where the fusing lysosomes release all luminal contents and merge fully with the plasma membrane. In contrast to this, receptor stimulation produces a small sustained calcium increase, which is associated with partial release of the lysosomal luminal content, and the lysosome membrane does not merge into the plasma membrane. In most cells, lysosomes express the synaptotagmin (Syt) isoform Syt VII; however, this isoform is not present on astrocyte lysosomes and exogenous expression of Syt VII on lysosome inhibits their exocytosis. Deletion of one of the most abundant Syt isoform in astrocyte--Syt XI--suppresses astrocyte lysosome exocytosis. This identifies lysosome as Syt XI-regulated exocytic vesicle in astrocytes. Further, inhibition of lysosome exocytosis (by Syt XI depletion or Syt VII expression) prevents repair of injured astrocytes. These results identify the lysosomes and Syt XI as the sub-cellular and molecular regulators, respectively of astrocyte cell membrane repair.


Utility and diagnostic accuracy of fallopian tube touch imprint cytology.
To determine the diagnostic accuracy of cytology smears in distinguishing between tube and non-tube structures. One hundred cytology smears of fallopian tube and non-tube structures (vessels, round and ovarian ligaments) were prepared from surgically removed uterus and fallopian tube specimens and stained by the Papanicolaou method. The slides were reviewed blindly by pathologists and interpreted as tube or non-tube structures. The results were compared to the histological examination of the same specimens. Results indicated an overall accuracy of 97% with a specificity of 98% and sensitivity of 96% for cytology smears, taking histology as the gold standard. Positive and negative predictive values were 96.1% and 97.9%, respectively. Cytology smears are a convenient and cost effective tool for laboratory confirmation of tubal sterilization. This method can reduce the costs of laboratory examination, especially in developing countries, where tubal sterilizations are done in large cohorts. However, histological slides remain the gold standard in cases of medicolegal problems.


Comparative potencies of amphetamine, fenfluramine and related compounds in taste aversion experiments in rats.
1 Rats failed to drink a flavoured solution when its consumption had been followed by injection of amphetamine (conditioned taste aversion).2 There was very little difference between the potencies of (+)- and (-)-amphetamine.3p-Chloromethamphetamine was a more potent aversive agent than methamphetamine.4 Strong taste aversions were also conditioned with other congeners of amphetamine. The rank order of potency was: fenfluramine > chlorphentermine >p-hydroxyamphetamine.5 Cocaine induced only moderate taste aversions, even at high doses.6 Aversive potency did not appear to be correlated with known neurochemical actions of the drugs or with behavioural stimulation, but appeared to be a central action which may have been linked to anorexigenic potency or time course of action.


Characterization of Brassica napus responses to diluted and undiluted industrial wastewater.
Rising water scarcity, together with increased industrial wastewater production, suggests reusing of wastewater for plant irrigation. The wastewater from Razi petrochemical complex contained different salts and heavy metals. Variation in Brassica napus responses to wastewater irrigation has recommended appropriate levels of mineral nutrients in diluted wastewater that stimulated plant growth, and toxic levels of salts in undiluted wastewater that restricted plant growth. The undiluted wastewater irrigation significantly decreased chlorophyll fluorescence, along with photosynthetic capacity, while wastewater dilution mitigated its adverse effect. High levels of salts in undiluted wastewater induced an imbalance in plant mineral nutrients, which was evidenced with increased lipid peroxidation and reduced plant growth. On the contrary to adverse effects of undiluted wastewater on plant performance, the diluted wastewater, especially at 50% level, behaved as a fertilizer which increased leaf mineral nutrients, photosynthetic capacity, morphological and anatomical features of plant, but decreased lipid peroxidation. In relation to improvement in photosynthetic capacity, a significant increase was achieved in stomatal traits in plants irrigated with half-strength wastewater. In conclusion, due to the nutrition values of wastewater, it can be suggested to irrigate plants with diluted wastewater with the aim of improving crop productivity and saving freshwater sources.


Presenteeism and Absenteeism Before and After Total Hip and Knee Arthroplasty.
Absenteeism is costly, yet evidence suggests that presenteeism-illness-related reduced productivity at work-is costlier. We quantified employed patients' presenteeism and absenteeism before and after total joint arthroplasty (TJA). We measured presenteeism (0-100 scale, 100 full performance) and absenteeism using the World Health Organization's Health and Work Performance Questionnaire before and after TJA among a convenience sample of employed patients. We captured detailed information about employment and job characteristics and evaluated how and among whom presenteeism and absenteeism improved. In total, 636 primary, unilateral TJA patients responded to an enrollment email, confirmed employment, and completed a preoperative survey (mean age: 62.1 years, 55.3% women). Full at-work performance was reported by 19.7%. Among 520 (81.8%) who responded to a 1-year follow-up, 473 (91.0%) were still employed, and 461 (88.7%) had resumed working. Among patients reporting at baseline and 1 year, average at-work performance improved from 80.7 to 89.4. A Wilcoxon signed-rank test indicated that postoperative performance was significantly higher than preoperative performance (P < .0001). The percentage of patients who reported full at-work performance increased from 20.9% to 36.8% (delta = 15.9%, 95% confidence interval = [10.0%, 21.9%], P < .0001). Presenteeism gains were concentrated among patients who reported declining work performance leading up to surgery. Average changes in absences were relatively small. Combined, the average monthly value lost by employers to presenteeism declined from 15.3% to 8.3% and to absenteeism from 16.9% to 15.5% (ie, mitigated loss of 8.4% of monthly value). Among employed patients before TJA, presenteeism and absenteeism were similarly costly. After, employed patients reported increased performance, concentrated among those with declining performance leading up to surgery.


Application of endoscope-assisted minimal-access techniques in orbitozygomatic complex, orbital floor, and frontal sinus fractures.
Minimal-access endoscope-assisted techniques have been adapted for facial rejuvenation surgery over the past 5 years, allowing smaller incisions, limited dissection, and shorter recovery times. Their increasing popularity prompted the author to investigate their application in facial trauma patients. Endoscope-assisted minimal-access surgical approaches were developed in cadavers and applied in the management of frontal sinus, orbital floor, and orbitozygomatic complex fractures over a 3-year period. At a Level 1 regional trauma center, these techniques were applied in 33 of 161 patients with facial fractures. Surgical techniques, applications of endoscope technology, and indications/contraindications are presented. A technique, known as the pulse test, was developed for assessing the integrity of the orbital floor. Endoscope-assisted minimal-access surgical techniques have a definite but perhaps limited application in the diagnosis, as well as in some aspects of the surgical treatment of fractures of the anterior table frontal sinus, orbitozygoma, and orbital floor. New indications for these techniques will depend upon the development of improved instrumentation.


Assay of the enantiomers of ibutilide and artilide using solid-phase extraction, derivatization, and achiral-chiral column-switching high-performance liquid chromatography.
Ibutilide fumarate and artilide fumarate, new drugs for the treatment of cardiac arrhythmias, each contain a stereogenic center bearing a secondary alcohol group. Reversed-phase achiral-chiral column-switching HPLC separations of the enantiomers of each compound were developed and validated for quantitation in plasma and other biofluids. The key component of the method was derivatization with 1-naphthyl isocyanate, which enhanced the sensitivity of fluorescence detection and enabled the enantiomers to be separated on a Pirkle column (covalent 3,5-dinitrobenzoyl-D-phenylglycine stationary phase). The lower limit of quantitation of ibutilide fumarate was typically 0.1 ng/ml or less of each enantiomer in 1 ml of plasma. Two of the special features of the column-switching system included operation with two samples in the system at one time, which reduced analysis time to 16 min/sample for ibutilide and 19 min/sample for artilide, and a relay-operated switching of detector outputs, which allowed achiral and chiral column chromatographic data to be gathered from two detectors into a single data acquisition channel.


The value of internal carotid systolic velocity ratio for assessing carotid artery stenosis with Doppler sonography.
The purpose of this study was to assess the value of the ratio between the internal carotid systolic velocity (ICSV) at the carotid bulb and the distal internal carotid systolic velocity (DICSV) for the detection of carotid artery stenosis. Two hundred eleven patients were enrolled in a prospective study comparing several Doppler measurements with carotid angiography. The following spectral measurements were obtained with Doppler sonography: ICSV/DICSV ratio, ICSV, ICSV/common carotid systolic velocity (CCSV) ratio, and internal carotid (end) diastolic velocity (ICDV). Receiver operating characteristic curves were generated for two groups: the first group being 365 carotid arteries for which all Doppler measurements were successfully obtained and the second being a subgroup of 258 carotid arteries for which an ICSV of 100 cm/sec or greater was present. In the group of 365 carotid arteries, the ICSV/DICSV ratio improved the detection of stenosis of 60% or greater as compared with the ICDV (p = .03). In 258 carotid arteries with an ICSV of 100 cm/sec or greater, the ICSV/DICSV ratio allowed for a better angiographic correlation for identifying stenoses of 60% or greater and 70% or greater as compared with ICSV (p = .006 and .023, respectively), ICSV/CCSV (p = .0013 and .003, respectively), and ICDV (p = .0015 and .020, respectively). Using the ICSV/DICSV ratio for the Doppler detection of carotid artery stenosis is advantageous when the ICSV is 100 cm/sec or greater.


Somatic inactivation of genes on chromosome 13 is a common event in retinoblastoma.
Through family studies and analysis of patients with congenital chromosome abnormalities, the germ-line mutation responsible for the hereditary form of the eye tumour, retinoblastoma, has been assigned to the q14 region on chromosome 13 and closely linked to an enzyme called esterase D (ESD). Knudson has proposed that as few as one somatic event in addition to the germ-line mutation is required to induce tumours in patients with the hereditary form of retinoblastoma; the non-hereditary form requires two somatic events to occur in the same cell. The somatic event(s) may involve either mutation of the remaining normal gene at 13q14 or mutation of a gene at another site in the genome. Here we have examined six retinoblastoma patients who are heterozygous for electrophoretic variants of ESD. Although the normal cells of all six patients expressed both variants, the tumour cells of four patients expressed enzyme from only one of the two ESD alleles. We tentatively conclude that induction of a retinoblastoma tumour requires the somatic inactivation of genes near the ESD locus including the remaining normal gene at the retinoblastoma (RB) locus.


Lipid metabolism and pro-oxidant/antioxidant balance of Halamphora oceanica from the Gulf of Mexico exposed to water accommodated fraction of Maya crude oil.
Diatoms play key roles in primary production and carbon fixation at a global scale and in some cases these species live on marine ecosystems impacted by crude oil (CO) spills. Halamphora oceanica, a new diatom species from the Southwest of the Gulf of Mexico was isolated and cultured in the laboratory and was exposed to water accommodated fraction (WAF) of different Maya CO loads at 0.01, 0.1, 1 and 10g/L by 96h. A battery of biomarkers involved in oxidative stress (O2•, H2O2, TBARS, ROOH, RC=O, SOD, CAT, GPx), biotransformation and conjugation (total CYP450 activity and GST) moreover fatty acid (FA) metabolism (FA levels, fatty-acid synthase and acyl-CoA oxidase) were measured. Obtained results suggest that increases of PAHs in the medium (below to EC50) acts as external forces able to turn-on regulatory mechanisms on H. oceanica involved in both, on the PAHs uptake and changing its aerobic metabolism to anaerobic metabolism. However, the growth of this microalgae species evaluated as chlorophyll "a" and pheophytin levels increased as the WAF concentration indicating that PAHs and other hydrosoluble hydrocarbons were used as carbon and energy sources by unidentified enzymes not evaluated in the current study. Our hypothesis was also corroborated by IBRv2. In the current study, we suppose the change from aerobic to anaerobic metabolism as a strategy for Halamphora oceanica survival exposed to petroleum hydrocarbons.


Chronic exposure to anxiolytic drugs, working by different mechanisms causes up-regulation of dihydropyridine binding sites on cultured bovine adrenal chromaffin cells.
Exposure of bovine adrenal chromaffin cells to ethanol [50 mM], alprazolam [10(-7) M] and buspirone [10(-7) M] inhibited basal and carbachol-induced release of catecholamines from these cells. The inhibition produced by alprazolam was prevented, and that produced by ethanol inhibited, by the presence of the benzodiazepine receptor antagonist, flumazenil [10(-8) M]. The inhibition produced by buspirone was unaffected by flumazenil, but was mimicked by the selective 5-HT1A receptor agonist, 8-OH DPAT and prevented by the 5-HT receptor antagonist spiperone [10(-6) M]. These results suggest that bovine adrenal chromaffin cells express GABAA receptors, containing a benzodiazepine recognition site and also 5-HT1A receptors. Ethanol and alprazolam appear to inhibit the excitability of bovine adrenal chromaffin cells by an action related to the former, while buspirone probably inhibits these cells through the latter. Maintaining bovine adrenal chromaffin cells for several days in culture medium, containing inhibitory concentrations of ethanol alprazolam or buspirone, produced a marked increase in binding sites for a [3H]dihydropyridine [DHP] calcium channel antagonist, on cell membranes. The increase in binding sites produced by alprazolam was greater than that produced by the other two agents and was almost completely prevented by the concomitant presence of flumazenil. The effects of ethanol and buspirone on the binding of DHP were not prevented by flumazenil. The results suggest that drugs which decrease excitability of bovine adrenal chromaffin cells by different mechanisms, may evoke a similar adaptive response involving an increase in DHP-sensitive calcium channels.


Studies on dispersed unincubated chick blastoderm cells. II. Formation of contacts and cell sorting in aggregates of unincubated chick blastoderm and heart cells.
The configuration of blastoderm-heart cell aggregates after cell sorting depended largely on the proportion of each cell type initially present in the culture. In heterotypic aggregates, blastoderm cells were always found to partially or completely surround heart cells with both cell types retaining their characteristic morphology. Specialized junctions (e.g. desmosomes) developed only between cells of the same type. These observations suggest that blastoderm cells are unable to recognize and form stable contacts with heart cells.


Room temperature benzene C-H activation by a new [PSiP]Ir pincer complex.
The synthesis and reactivity of coordinatively unsaturated Rh and Ir complexes supported by the new bis(phosphino)silyl pincer ligand [kappa(3)-(2-Cy(2)PC(6)H(4))(2)SiMe](-) ([Cy-PSiP](-)) are reported, including the first example of facile, room temperature intermolecular arene C-H bond activation mediated by a silyl pincer complex.


Assessment of the quality of hospital care for children in Indonesia.
To obtain an overview of the quality of care for children in Indonesia, by assessing hospitals with a view to proceed to a quality improvement mechanism for child care. Stratified two-stage random sampling in six regions identified 18 hospitals (provinces Jambi, East Java, Central Kalimantan, South-East Sulawesi, East Nusa Tenggara, North Maluku). Three randomly selected hospitals in each province were visited by trained assessors who scored each assessed service (expressed as a percentage of achievement) and grouped into good (≥ 80%), requiring improvement (60-79%) and urgently requiring improvement (< 60%). The overall median result score across all areas was 43% (IQR 28%-53%). Case management for common childhood illnesses had a median score of 37% (IQR18-43%), neonatal care 46% (IQR 26-57%) and patient monitoring 40% (IQR 30-50%), all indicating an urgent need for improvement. Qualitative data showed as main problems inadequate use of standard treatment guidelines, irrational prescribing of antibiotics, poor progress monitoring and poor supportive care. We found serious shortcomings in the quality of hospital care for children. Finding and documenting those is the first step in a quality improvement process. Work is needed to start an improvement cycle for hospital care.


Histone deacetylase inhibitors suppress CXCR4-mediated dendritic cell migration by regulation of maturation process.
Once activated by an infected pathogen, dendritic cells (DC's) migrate toward secondary lymphoid organs, and release inflammatory mediators. Therefore, in some case, mature DC's (mDC's) are considered to be potent inflammatory inducers. In this study we demonstrated that histone acetylation plays an important regulatory role in conserving the migration activity of the DC's. We showed that histone deacetylase (HDAC) inhibition reduces CXC chemokine receptor 4 (CXCR4)-dependent DC's migration. These inhibitory effects were found to be caused by a reduction in the expression of CXCR4, and by the phosphorylation of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases 1 and 2 (ERK 1/2), and c-Jun N-terminal kinase (JNK). Taken together, histone deacetylase inhibitors (HDACi's) inhibit the phosphorylation of MAP kinases, and this inhibition reduces the expression of CXCR4, and this reduction decreases the chemotactic activity of mDC's.