PLASMA CAPILLARY ZONE ELECTROPHORESIS AND PLASMA CHEMISTRY ANALYTES IN TORTOISES (TESTUDO HERMANNI, TESTUDO GRAECA) AND TURTLES (TRACHEMYS SCRIPTA ELEGANS, GRAPTEMYS SPP.) IN FALL.
Clinical health checks including blood testing before brumation in fall are an important tool in reptile medicine and help to reduce the risk of complications during brumation. Electrophoresis may be useful for the evaluation of liver cell function and the detection of antigenic stimulation. The goal of this study was to compare species-specific reference intervals for plasma chemistry analytes and capillary zone electrophoresis (CZE) for a variety of pet chelonian species in fall. Lithium heparinized samples were collected from 68 clinically healthy spur-thighed tortoises (Testudo graeca), 79 red-eared sliders (Trachemys scripta elegans), and 30 map turtles (Graptemys spp.) from September to November 2016 and 2017. During the same period, 128 equivalent samples were collected from Hermann's tortoises (Testudo hermanni) and the obtained data was used for comparison. Chemistry analytes were measured from plasma using an autoanalyzer and electrophoresis was carried out by CZE. Significant variations (P < 0.0001) between the species were found for several analytes including total protein, percent albumin, and albumin: globulin ratio, α-, β-, and γ-globulin, alkaline phosphatase, glutamate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, bile acids, creatine kinase, urea, uric acid, inorganic phosphorus, total calcium, and potassium. The variations in albumin (g/l) measured by CZE were also significant (P = 0.0064). No significant variations were detected for sodium levels. The results demonstrate the importance of species-specific reference intervals and provide reference intervals for the plasma chemistry and capillary zone electrophoresis in three chelonian species.


The effects of altered sterol composition on the mitochondrial adenine nucleotide transporter of Saccharomyces cerevisiae.
1. The membrane sterol composition of mitochondria of the ole-3 mutant of Saccharomyces cerevisiae was manipulated by growing the organism in the presence of Tween 80 (1%, W/V) plus defined supplements o- delta-aminolaevulinate. 2. Changes in mitochondrial sterol content induced considerable changes in the adenine nucleotide transporter. 3. As the sterol content was decreased, the affinity of the transporter for ATP did not alter significantly, but the rate of ATP uptake was greatly decreased, the total number of atractylate-sensitive binding sites diminished, and the proportion of high-affinity binding sites was decreased. 4. Since sterol depletion also uncouples oxidative phosphorylation [Astin & Haslam (1977) Biochem. J., 166, 287-298] and prevents the intramitochondrial generation of ATP, the decrease in the rate of ATP uptake by sterol-depleted mitochondria will cause a decrease in intramitochondrial ATP concentrations in vivo. This probably explains the inhibition of mitochondrial macromolecular synthesis that has previously been reported in lipid-depleted yeast mitochondria.


Weight gain and comorbidities associated with oral second-generation antipsychotics: analysis of real-world data for patients with schizophrenia or bipolar I disorder.
Many second-generation antipsychotics (SGAs) are associated with weight gain and cardiometabolic effects. Antipsychotic-associated weight gain is linked to treatment interruptions, potentially increasing risk of relapse and hospitalization. This retrospective study assessed clinically significant weight gain (CSWG), treatment interruptions, and development of cardiometabolic conditions in patients with schizophrenia (SZ) or bipolar I disorder (BD-I) following initiation of oral SGAs with moderate to high weight gain risk. Patients with no prior use of moderate to high weight gain risk oral SGAs were identified from patient-level medical/pharmacy claims and electronic medical records (January 2013-February 2020; OM1 Real-World Data Cloud). Those with ≥ 1 weight measurement in both the 12 months preceding and 3 months after SGA initiation (index date) were analyzed for continuous changes in weight, CSWG (≥ 7% and ≥ 10% increases from baseline), treatment interruptions (switches/discontinuations), and development of cardiometabolic conditions. Median follow-up times in the SZ (n = 8174) and BD-I (n = 9142) cohorts were 153.4 and 159.4 weeks, respectively; 45.5% and 50.7% were obese at baseline. Mean (SD) percent weight increase during treatment was 3.3% (7.2) and 3.7% (7.0) for patients with SZ and BD-I, respectively, and was highest for underweight/normal weight patients (SZ: 4.8% [8.1]; BD-I: 5.5% [8.7]). More than 96% had treatment interruptions during follow-up, primarily discontinuations. CSWG and treatment interruptions occurred within a median of 13 and 14 weeks after treatment initiation, respectively. Of patients with CSWG and treatment interruptions, approximately 75% did not return to baseline weight during follow-up. Among those without baseline cardiometabolic conditions, 14.7% and 11.3% of patients with SZ or BD-I, respectively, developed ≥ 1 condition over 12 months post-index. Incidence was generally highest among those who were overweight/obese at baseline and those who experienced CSWG. In this analysis of real-world data, both weight gain and treatment interruptions occurred early in treatment for patients with SZ or BD-I. Treatment-associated weight gain persisted despite switching or discontinuing index treatment. Additionally, cardiometabolic morbidity increased within 12 months of treatment initiation. Patients with SZ or BD-I are at greater risk than the general population for cardiometabolic conditions; weight gain associated with SGAs may exacerbate these health risks.


Analysis of the Localization of Fluorescent PpROP1 and PpROP-GEF4 Fusion Proteins in Moss Protonemata Based on Genomic "Knock-In" and Estradiol-Titratable Expression.
Tip growth of pollen tubes, root hairs, and apical cells of moss protonemata is controlled by ROP (Rho of plants) GTPases, which were shown to accumulate at the apical plasma membrane of these cells. However, most ROP localization patterns reported in the literature are based on fluorescent protein tagging and need to be interpreted with caution, as ROP fusion proteins were generally overexpressed at undefined levels, in many cases without assessing effects on tip growth. ROP-GEFs, important regulators of ROP activity, were also described to accumulate at the apical plasma membrane during tip growth. However, to date only the localization of fluorescent ROP-GEF fusion proteins strongly overexpressed using highly active promoters have been investigated. Here, the intracellular distributions of fluorescent PpROP1 and PpROP-GEF4 fusion proteins expressed at essentially endogenous levels in apical cells of Physcomitrella patens "knock-in" protonemata were analyzed. Whereas PpROP-GEF4 was found to associate with a small apical plasma membrane domain, PpROP1 expression was below the detection limit. Estradiol-titratable expression of a fluorescent PpROP1 fusion protein at the lowest detectable level, at which plant development was only marginally affected, was therefore employed to show that PpROP1 also accumulates at the apical plasma membrane, although within a substantially larger domain. Interestingly, RNA-Seq data indicated that the majority of all genes active in protonemata are expressed at lower levels than PpROP1, suggesting that estradiol-titratable expression may represent an important alternative to "knock-in" based analysis of the intracellular distribution of fluorescent fusion proteins in protonemal cells.


High affinity binding of [3H]acetylcholine to muscarinic receptors. Regional distribution and modulation by guanine nucleotides.
The interaction of [3H]acetylcholine ([3H]AcCh) with the muscarinic receptor was studied in seven distinct rat brain regions and in heart atrium by employing 10 microM atropine to define specific binding. The specific binding exhibited by the labeled neurotransmitter was found to be sensitive to muscarinic but not to nicotinic drugs. The muscarinic high affinity agonist-binding sites were characterized with respect to their binding properties, regional distribution, pharmacology, and modulation by guanyl nucleotides and by transition metal ions. In all tissues examined, specific binding of [3H]AcCh was saturable over the range of 4-200 nM and occurred in a receptor population that was apparently homogeneous and had a dissociation constant of approximately 19-39 nM in most of the regions. The ratio of muscarinic receptors labeled by [3H]AcCh to those labeled by the potent antagonist [3H]N-methyl-4-piperidylbenzilate varied markedly among tissues, from 0.15 in the hippocampus to 0.71 in the atrium. This ratio was lower in brain regions rich in muscarinic receptors, where smaller sensitivity of [3H]AcCh binding to guanyl nucleotides was also observed. In the presence of the latter [3H]AcCh binding was decreased by 25 to 90% in different tissues, with the greatest decreases occurring in the atrium and brainstem. In the latter preparations, transition metal ions do not affect [3H] AcCh binding, while in the other preparations studied they induce an increase in the binding capacity for the labeled neurotransmitter, which is sensitive to guanine nucleotides.


Alveolar hydatid disease. A review of clinical features of 33 indigenous cases of Echinococcus multilocularis infection in Alaskan Eskimos.
The clinical features of 33 cases of alveolar hydatid disease (AHD) in Alaskan Eskimos and a review of the surgical experience with this disease are presented. Among untreated patients, progression of the disease to a fatal outcome was observed in 70%. The primary hepatic lesion resembles cancer, and errors in diagnosis by both the surgeon and pathologist are common. Although surgical resection of the entire primary hepatic lesion offers the only proven curative treatment, only 26% of those explored were resectable. All seven patients resected for cure are alive 6-27 years post-operatively (average survival, 14.7 years). A 5-year experience with continuous mebendazole therapy in the management of five nonresectable cases of AHD indicates that a favorable effect of this drug is being observed. It now appears that Echinococcus infections are no longer the sole province of the surgeon. Although the role of medical therapy is not yet clearly defined, it must be considered in the management of all cases of AHD. The first reported locally-acquired case of AHD in the conterminous United States, and the widespread occurrence and expanding range of E. multilocularis in the north-central United States and south-central Canada, point to the increasing public health importance of alveolar hydatid disease.


Archaeal type III RuBisCOs function in a pathway for AMP metabolism.
The type III ribulose-1,5-bisphosphate carboxylase-oxygenase (RuBisCO) present in the archaeon Thermococcus kodakaraensis was found to participate in adenosine 5'-monophosphate (AMP) metabolism, a role that is distinct from that of classical RuBisCOs of the Calvin-Benson-Bassham cycle. Genes annotated as thymidine phosphorylase (deoA) and eucaryal translation initiation factor 2B (e2b2) were found to encode AMP phosphorylase and ribose-1,5-bisphosphate isomerase, respectively. These enzymes supplied the RuBisCO substrate, ribulose-1,5-bisphosphate, from AMP and phosphate. Archaea with type III RuBisCOs all harbor both DeoA and the corresponding E2b2 homologs. In this pathway, adenine was released from AMP and the phosphoribose moiety entered central-carbon metabolism.


Early metabolic imprinting events increase marbling scores in fed cattle.
Early weaning of calves to a high concentrate diet results in greater fat deposition and suggests early postnatal metabolic imprinting events may be exploited as a management tool to improve cattle value. Our objective was to implement a short, high energy dietary intervention before a typical grazing period to manipulate intramuscular fat deposition in finishing cattle. Fall-born, Angus-sired steer calves (n = 24) were stratified by sire and randomly assigned to normal weaned (NW) or metabolic-imprinted (MIP) treatments. At 105 ± 6d (135kg), MIP calves were transitioned to a diet containing 20% CP and 1.26 Mcal/kg NEg. Metabolic-imprinted calves were fed ad libitum as a group. Normal weaned calves remained on their dam until 253 ± 6 d of age. At this time, treatment groups were combined and grazed for 156 d on a mixed summer pasture. Following the grazing phase, steers were adapted to a corn silage-based feedlot diet and performance was monitored on 28-d intervals. Calves were staged for harvest based on backfat endpoint (target 1.0 to 1.2 cm). Metabolic-imprinted calves were heavier (P < 0.05) than NW calves (341 vs. 265 ± 4.2 kg) at normal weaning age. During the grazing phase, NW steers gained more weight than (P < 0.05) MIP steers (0.69 vs. 0.35 ± 0.03 kg/d). Feedlot performance and USDA yield grade were similar (P > 0.20) between treatments. However, MIP steers produced heavier (P < 0.05) carcasses (564 vs. 524 ± 5.6 kg) with higher (P < 0.001) marbling scores (645 vs. 517 ± 23). Therefore, calves consuming a high concentrate diet for 148 d after early weaning produced higher quality carcasses. This suggests early weaning and feeding a high concentrate before grazing is a viable strategy to increase marbling deposition compared with a traditional production system.


[Neuropathological principles of behavioral disorders in dementia].
Psychotic symptoms, apathy, agitation and aggressiveness are behavioral disorders that occur frequently in patients with Alzheimer's dementia. They cause serious problems for patients, relatives and care-givers. These behavioral disorders are associated with neuropathologic changes and alterations of brain metabolism in specific brain areas. Disturbances in mesotemporal and frontal brain areas seem to be related to psychotic symptoms. Apathy is associated with dysfunction of frontal cortical areas. Agitation and impulsivity appear to result from a hypofunction of the serotonin system in association with a relative hyperfunction of dopaminergic and noradrenergic systems. These dysfunctions are the result of direct neuropathologic changes, but also due to cholinergic deficits that seem to both contribute synergistically to and independently cause behavioral disturbances.


Methylenetetrahydrofolate reductase polymorphisms and the risk of gestational hypertension.
Evidence on the association of 5,10 methylentetrahydrofolate reductase (MTHFR) C677T and A1298C polymorphisms in women with gestational hypertension is inconsistent. It is also unknown whether the fetal genotype is relevant, or whether folic acid supplementation modifies this association. The study population was composed of U.S. and Canadian white women with nonmalformed infants participating in the Slone Epidemiology Center Birth Defects Study between 1993 and 2000. Women were interviewed within 6 months after delivery regarding multivitamin use in pregnancy and the occurrence of gestational hypertension, among other factors. DNA was extracted from cheek swabs and gene alleles determined by restriction fragment length polymorphism analysis. We compared the prevalence of the 677TT/CT and 1298CC/AC genotypes between cases with gestational hypertension (54 mothers and their 51 offspring) and controls (100 mothers and their 99 offspring). We also estimated odds ratios (ORs) and 95% confidence intervals (CIs) using conditional logistic regression to control for geographic region and calendar year. The T allele was present in 69% of women with gestational hypertension versus 57% of control women (compared with 677CC, OR = 1.9; 95% CI = 0.9-4.0). The offspring of case and control women had a 677TT/CT genotype prevalence of 68% and 47%, respectively (2.4; 1.1-5.0). Among women supplemented with folic acid during the first 5 months of pregnancy, the ORs for maternal and fetal 677TT/CT genotypes were 0.9 (0.3-2.5) and 2.1 (0.7-6.0), respectively. Neither maternal nor fetal 1298CC/AC genotypes were associated with an increased risk of gestational hypertension. Maternal and fetal MTHFR C677T polymorphism may be associated with a moderately increased risk of gestational hypertension, and there is a suggestion that this association may be diminished among women receiving folate supplementation during pregnancy.


[Results of retinalamine use in the treatment of tuberculous chorioretinitis].
The efficiency of the peptide bioregulator retinalamine in the complex treatment of patients with tuberculous chorioretinitis was studied. Ninety-seven patients (69 females and 28 males) with inactive tuberculous chorioretinitis (132 eyes) were followed up. According to the treatment option, the patients were divided into 2 groups: a study group and a control one. Parabulbar injections of retinalamine were used in the study group. The conventional treatment using parabulbar injections of emoxipine (an agent from the group of synthetic antioxidants) was performed in the control group. The results of treatment were assessed by visual acuity, the number of pathological scotomas in the field of vision and the findings of electrophysiological investigations (EPI). Analysis of the results of conventional versus peptide bioregulator therapy used in patients with inactive tuberculous chorioretinitis showed the advantages of retinalamine. This manifested itself as increased visual acuity in 66.3% of the patients receiving retinalamine while in the emoxipine-treated patients, visual acuity increased only in 20% of cases. In the patients taking retinalamine, the positive changes in the field of vision were statistically significant in the number of normal test objects, relative and absolute scotomas while in the emoxipine group, only the reduction in the number of absolute scotomas was significant. The values of EPI suggest that the peptide bioregulator is more effective in enhancing the functional activity of retinal neurons than the synthetic antioxidant.


[Pathogenetic mechanisms of phantom-pain syndrome].
This review considers the literature data on the epidemiology of phantom-pain syndrome (PPS) presents the results of numerous clinical studies demonstrating the lack of effectiveness of the vast majority of modem non-pharmacological and pharmacological methods of treatment of PPS. Detail presents data on the patho genetic mechanisms underlying the PPS. According to most researchers, the major role in the patho genesis of the PPS has the reorganization of the somatosensory area of the cerebral cortex of the brain. At the same time discusses the views of researchers who believe that the main reason PPS is to strengthen nociceptive and nonnociceptive afferentation in the peripheral newous system. The comparison of these conflicting data it is concluded that in the genesis of the PPS plays the role of both primary and secondary sensitization. Leading important dysfunction of the central nervous system. Details the modern understanding of the mechanisms underlying the high efficiency of suppression of PPS during stimulation of motor cortex.


Thinking on your feet: Anticipatory foot placements in repeated bimanual object displacements.
Effective handling of objects requires proper use of the hands. If the object handling is done while standing or walking, it also requires proper use of the feet. We asked how people position their feet to meet future and ongoing object-handling demands. In previous research on this topic, participants walked to a table and picked up an object for a single displacement from one place to another. These studies shed light on sensitivity to kinematics but, strictly speaking, may not have revealed anything about sensitivity to dynamics. In the present study, we asked participants to walk to a table to move an object back and forth over different distances and at different rates. Prior to walking to the table, participants had full knowledge of what the task would be. By using a rhythmic rather than discrete object placement task, we could analyze participants' sensitivity to dynamics as well as kinematics. Consistent with our expectation that participants would tune their foot separations to demands related to dynamics, we found that stance width was wider for long than for short object displacements and was more pronounced for high displacement rates than for low displacement rates. Also consistent with our expectations about planning, these effects were evident as soon as participants reached the table. Our results add to the limited research on coordinated action of the hands and feet in purposeful object manipulation.


Factor XIIIA and clot strength after cardiopulmonary bypass.
Reduced factor XIIIA levels and decreased clot strength have been associated with increased bleeding after cardiopulmonary bypass (CPB). The purpose of this study was to evaluate the relationship between hemostatic factors, including factor XIIIA, and clot strength before, during and after CPB. Factor XIIIA antigen, platelet counts, fibrinogen, factor V activity, tissue plasminogen activator and clot strength (by thromboelastograph) were measured at baseline, after 45 min of CPB, at the end of CPB and 4 h post-operatively in 34 patients. Baseline factor XIIIA antigen was 5.2 +/- 1.4 mg/l. On average, factor XIIIA levels dropped to 64% and clot strength to 77% of baseline values after 45 min on CPB and remained below baseline during the immediate post-operative period. Clot strength was significantly correlated (r = 0.81) with platelet count and fibrinogen but not plasma factor XIIIA levels. Addition of 10 mg/l recombinant factor XIII[a2] significantly increased clot strength. Postoperative bleeding at 2 h was inversely correlated with platelet count, factor XIIIA antigen and clot strength measured at the end of CPB. Maintenance of adequate platelet counts and factor XIIIA levels at the end of CPB may play a role in maintaining clot strength and reducing blood loss.


Priority setting of ICU resources in an influenza pandemic: a qualitative study of the Canadian public's perspectives.
Pandemic influenza may exacerbate existing scarcity of life-saving medical resources. As a result, decision-makers may be faced with making tough choices about who will receive care and who will have to wait or go without. Although previous studies have explored ethical issues in priority setting from the perspective of clinicians and policymakers, there has been little investigation into how the public views priority setting during a pandemic influenza, in particular related to intensive care resources. To bridge this gap, we conducted three public town hall meetings across Canada to explore Canadian's perspectives on this ethical challenge. Town hall discussions group discussions were digitally recorded, transcribed, and analyzed using thematic analysis. Six interrelated themes emerged from the town hall discussions related to: ethical and empirical starting points for deliberation; criteria for setting priorities; pre-crisis planning; in-crisis decision-making; the need for public deliberation and input; and participants' deliberative struggle with the ethical issues. Our findings underscore the importance of public consultation in pandemic planning for sustaining public trust in a public health emergency. Participants appreciated the empirical and ethical uncertainty of decision-making in an influenza pandemic and demonstrated nuanced ethical reasoning about priority setting of intensive care resources in an influenza pandemic. Policymakers may benefit from a better understanding the public's empirical and ethical 'starting points' in developing effective pandemic plans.


Practical non-pharmacological intervention approaches for sleep problems among older adults.
Poor sleep is common among older adults, often caused by multiple underlying factors such as chronic stress. Poor sleep is subsequently associated with negative health outcomes including higher morbidity and mortality. Our primary purpose is to explore practical non-pharmacological intervention approaches integrating stress management to improve sleep quality among older adults. In doing so, we highlight approaches that appear to hold promise in real-world settings with older individuals. We conducted a tailored literature review specifically on approaches to improve sleep quality among older adults, with emphasis on those integrating stress management. Online search engines were reviewed to identify research in these areas. Various non-pharmacological intervention approaches, such as mindfulness and cognitive behavioral therapy, have shown promise in improving sleep quality and health outcomes within this population. Those integrating chronic stress management appear to be particularly successful. Thus further development of multidimensional sleep interventions integrating stress management with seniors is warranted.


Analysis of recombinant dengue virus polypeptides for dengue diagnosis and evaluation of the humoral immune response.
Dengue is a serious cause of morbidity and mortality in tropical areas worldwide. We cloned and expressed recombinant polypeptides spanning the entire genome of a Brazilian dengue virus type 2 (DENV-2) strain in contiguous segments to generate antigens for dengue diagnosis and evaluation of the human humoral immune response. When analyzed by Western blot and an enzyme-linked immunosorbent assay (ELISA) using human sera, the most reactive polypeptide (pD2-3(E)) was located in the N-terminal portion of the envelope protein. The sensitivity of an IgG-ELISA using pD2-3(E) versus mouse brain antigen was 100% with convalescent sera and 79% with acute sera, with a specificity of 100%. Sera from patients infected with other DENV serotypes recognized pD2-3(E) equally well, whereas sera positive for yellow fever, rubella, and measles showed little or no reactivity. Using this novel approach, we identified a candidate antigen to facilitate diagnosis of DENV infections and observed a surprising variability in antibody patterns in the clinical response to DENV infections.


The F box protein partner of paired regulates stability of Drosophila centromeric histone H3, CenH3(CID).
Centromere identity and function is determined by the specific localization of CenH3 (reviewed in [1-7]). Several mechanisms regulate centromeric CenH3 localization, including proteasome-mediated degradation that, both in budding yeast and Drosophila, regulates CenH3 levels and prevents promiscuous misincorporation throughout chromatin [8, 9]. CenH3(CENP-A) proteolysis has also been reported in senescent human cells [10] or upon infection with herpes simplex virus 1 [11]. Little is known, however, about the actual mechanisms that regulate CenH3 proteolysis. Recent work in budding yeast identified Psh1 as an E3-ubiquitin ligase that mediates degradation of CenH3(Cse4p) [12, 13], but E3-ligases regulating CenH3 stability in metazoans are unknown. Here, we report that the F box protein partner of paired (Ppa), which is a variable subunit of the main E3-ligase SCF [14-17], mediates CenH3(CID) stability in Drosophila. Our results show that Ppa depletion results in increased CenH3(CID) levels. Ppa physically interacts with CenH3(CID) through the CATD(CID) that, in the fly, mediates Ppa-dependent CenH3(CID) stability. Altogether, these results strongly suggest that, in Drosophila, SCF(Ppa) regulates CenH3(CID) proteolysis. Interestingly, most known SCF complexes are inactive when, at mitosis, de novo CenH3(CID) deposition takes place at centromeres, suggesting that, in Drosophila, CenH3(CID) deposition and proteolysis are synchronized events.


Intravenous cangrelor as a peri-procedural bridge with applied uses in ischemic events.
Cangrelor is a relatively new antiplatelet drug that has been approved for use as an adjunct therapy to percutaneous coronary intervention (PCI) to decrease peri-procedural myocardial infarction (MI), coronary revascularization, and stent thrombosis. Cangrelor is an adenosine triphosphate analogue with a pharmacokinetic mechanism based on a reversible, dose-dependent inhibition adenosine diphosphate (ADP)-induced platelet aggregation. This drug has lately been in the spotlight as a possible bridge therapy for anti-platelet medication prior to cardiac and non-cardiac surgeries. Platelet function is usually restored within sixty minutes of cessation of therapy, thereby decreasing the risk of bleeding while providing adequate pre-procedural coverage to reduce ischemic events. This manuscript reviews the literature on cangrelor and summarizes its role as a peri-procedural bridge.


Self-reported health and driving cessation in community-dwelling older drivers.
Stopping driving has significant negative consequences for older adults, but there is no simple, reliable screening tool to predict driving cessation. We sought to determine if self-rated health (SRH) was an independent predictor of driving cessation among older adults. Data on SRH (poor, fair, good, very good, or excellent), medical diagnoses, physical performance, visual acuity, driving status, and other relevant covariates were collected from 649 community-dwelling older Alabama drivers during in-home interviews. Using multivariable logistic regression analyses, we estimated the association of SRH with driving cessation 2 years later. Participants had a mean age of 74 years; 43% were women, 41% African American, and 48% rural. Overall, 36% reported poor to fair SRH at baseline, and 11% had stopped driving after 2 years. Compared to 8% of drivers with good to excellent SRH, 17% with poor to fair health stopped driving (adjusted odds ratio [OR], 1.93; 95% confidence interval [CI], 1.09-3.41; p=.025). Lower Short Physical Performance Battery (SPPB) scores (adjusted OR, 0.86; 95% CI, 0.78-0.95; p=.001) and older age (adjusted OR, 1.06 per year; 95% CI, 1.01-1.11; p=.010) were also associated with driving cessation. Receiver operating characteristics curves documented similar predictive discrimination (c statistics) for SRH (0.72), the SPPB (0.70), and a count of comorbidities based on the Charlson Comorbidity Index (0.73). Poor to fair SRH predicted incident driving cessation after 2 years in a cohort of older adults. SRH can be easily obtained during clinic visits to identify at-risk drivers.


How clinical practitioners assess frailty in their daily practice: an international survey.
Various operational definitions have been proposed to assess the frailty condition among older individuals. Our objective was to assess how practitioners measure the geriatric syndrome of frailty in their daily routine. An online survey was sent to national geriatric societies affiliated to the European Union Geriatric Medicine Society (EUGMS) and to members of the European Society for Clinical and Economic Aspects of Osteoporosis, Osteoarthritis and Musculoskeletal Diseases (ESCEO). A total of 388 clinicians from 44 countries answered to the survey. Most of them were medical doctors (93%), and their primary field of practice was geriatrics (83%). Two hundred and five clinicians (52.8%) always assessed frailty in their daily practice, 38.1% reported to "sometimes" measure it, and 9.1% never assess it. A substantial proportion of clinicians (64.9%) diagnose frailty using more than one instrument. The most widely used tool was the gait speed test, adopted by 43.8% of the clinicians, followed by clinical frailty scale (34.3%), the SPPB test (30.2%), the frailty phenotype (26.8%) and the frailty index (16.8%). A variety of tools is used to assess frailty of older patients in clinical practice highlighting the need for standardisation and guidelines.


Rational Design of a Highly Dispersed Fe-N-C Nanosheet with 1,10-Phenanthroline-2,9-Dicarboxylic Acid as a Preorganized Ligand: Boosted Electrochemiluminescence Detection of Tetracycline.
In view of the shortcomings of the current coreactant electrochemiluminescence (ECL) and inspired by natural oxygen (O2) reduction metalloenzymes, a novel ECL amplification strategy was established. A pyrolytic iron- and nitrogen-doped (Fe-N-C) nanosheet rich in singly ionized oxygen vacancy (VO•) defects was rationally designed by destroying the highly saturated coordination with a preorganized ligand 1,10-phenanthroline-2,9-dicarboxylic acid (PDA). Extraordinary catalytic activity for O2 activation was obtained via screening a special pyrolysis temperature using spectroscopic and electrochemical methods. The high-spin ferric centers of highly dispersed FeC nanoclusters and abundant carbon and oxygen vacancy defects fully contributed to the inherent catalytic activity. ECL amplification was achieved by integrating the material with luminol to generate redox-active radicals in situ from dissolved O2 and simultaneously shorten the transferring distance of radicals. Tetracycline (TC), which posed a growing threat to aquatic biodiversity and environmental safety, as a model antibiotic was successfully detected with a detection limit of 3.88 nM (S/N = 3), clarifying a promising application prospect of this new effective ECL amplification strategy in biological analysis and environmental monitoring.


Analysis of community composition during moderately thermophilic bioleaching of pyrite, arsenical pyrite, and chalcopyrite.
An analysis of the community composition of three previously undefined mixed cultures of moderately thermophilic bioleaching bacteria grown at 45 degrees C on pyrite, arsenical pyrite, and chalcopyrite has been carried out. The bacterial species present were identified by comparative sequence analysis of the 16S rRNA gene isolated from the bioleaching vessels and analyzed by denaturing gradient gel electrophoresis, cloning, and sequencing. The mixed cultures leached all three minerals, as shown by the increase in iron released from the mineral concentrates. The species identified from the mixed cultures during bioleaching of pyrite, arsenical pyrite, and chalcopyrite were clones closely related to Acidithiobacillus caldus C-SH12, Sulfobacillus thermosulfidooxidans AT-1, " Sulfobacillus montserratensis" L15, and an uncultured thermal soil bacterium YNP. It was also found that the same mixed culture maintained for over a year on chalcopyrite mineral selected approximately the same consortia of bacteria as the original mixed culture grown on chalcopyrite.


Current Perspectives on the Role of Very-Low-Energy Diets in the Treatment of Obesity and Type 2 Diabetes in Youth.
In both developed and developing countries, pediatric obesity and type 2 diabetes are an increasing public health concern: globally 5.6% of girls and 7.8% of boys aged ≥5 years have obesity. The incidence of type 2 diabetes has increased in youth in recent decades and disproportionately affects those from ethnic/racial minority groups and disadvantaged backgrounds. For the treatment of both conditions, conventional lifestyle intervention is frequently ineffective, access to bariatric surgery is very limited and many young people are unsuitable or unwilling to undergo surgery. A very-low-energy diet (VLED) provides a viable alternative and may be effective for weight reduction and improved glycemic control in youth, based on one systematic review. In particular, in the treatment of type 2 diabetes, a chart review and a pilot study both demonstrated that a VLED can reduce the requirement for medications, including insulin, and lead to the remission of diabetes. However, long-term follow-up and safety data remain limited and therefore a VLED is inconsistently recommended by clinical practice guidelines for the treatment of pediatric obesity and type 2 diabetes. In clinical practice, VLED use in children and adolescents is uniquely challenging due to intolerance of expected side effects, difficulty adhering to the highly restrictive diet and difficulty with behaviour change within the current social context and environment. Ultimately, more research, including larger, longer-term trials with comprehensive safety monitoring are required to strengthen the evidence base. This would inform clinical practice guidelines, which may facilitate more widespread utilization of VLED programs in the management of obesity and type 2 diabetes in youth.


Mesangial cell signaling cascades in response to mechanical strain and glucose.
Elevated glucose levels and glomerular hypertension (Pgc) are considered to contribute to the elaboration of matrix protein by mesangial cells (MCs) in diabetic glomeruli. MCs grown in 30 mM of glucose produce excessive matrix protein, as do MCs exposed to cyclic strain, and the combination of the two exacerbates this. Tight glucose control or reduction in Pgc clinically delays progression of diabetic nephropathy. MC c-fos is induced in response to either application of strain or high ambient glucose, inducing increases in activated protein-1 transactivational activity and extracellular matrix production. Stimuli that lead to c-fos induction pass through the three mitogen-activated protein (MAP) kinase pathways: p44/42, SAPK/JNK, and p38/HOG. We studied MAP kinase activation in MCs exposed to mechanical strain and a high glucose. MCs (passage 5 through 10) cultured for 96 hours on type 1 collagen-coated flexible-bottom plates in either 5.6 or 30 mM glucose were exposed to 5, 10, or 30 minutes of cyclic strain (60 cycles per min) by computer-driven generation of vacuums of -14 kPa, inducing 20% elongation in the diameter of the surface. Control MCs were grown on both coated rigid and flexible-bottom plates. Protein levels (by Western blot) and activity assays for all three kinase cascades were performed at baseline and after 5, 10, and 30 minutes. All experiments were performed in triplicate. MAP kinase signaling was seen in response to stretch, and high ambient glucose affected the pattern of activation. Both p44/42 and p38HOG kinase activities showed small increases to a maximum of 2.5- to 3.5-fold greater than static MCs at 10 minutes. Activity in both kinase cascades was slightly suppressed by 30 mM glucose. In contrast, SAPK/JNK activity was present at a very low level in static MCs and increased markedly by 10 minutes of stretch. Thirty micromolars of glucose augmented this effect by a factor of six over MCs cultured in 5.6 mM glucose after 10 minutes of stretch. Neither glucose concentration nor mechanical strain had any effect on the protein expression of any of the kinases by Western blot. MAP kinase cascade signaling is seen when physical force is applied to MCs, and glucose affects the pattern of activity. Thirty micromolars of glucose markedly increase the level of SAPK/JNK activation. This may have implications in diabetic signal transduction and matrix protein production.


Precision of Epithelial Defect Measurements.
To ensure optimal care of patients, cornea specialists measure corneal features, including epithelial defects (ED), with slit-lamp calipers. However, caliper measurements are subject to interphysician variability. We examined the extent of variability in ED measurements between cornea specialists and discuss the potential clinical impact. A total of 48 variably sized EDs were created in pig eyes. Three cornea specialists measured the maximum vertical and horizontal ED lengths to the nearest 10th of a millimeter using slit-lamp microscopy. An absolute difference in ED measurement between cornea specialists of 0.5 mm was chosen to be the a priori threshold for clinical significance and was evaluated by the Wilcoxon signed-rank test. Interrater reliability was assessed by intraclass correlation coefficients. The average absolute difference in the vertical ED length between pairs of examiners ranged from 0.54 to 0.63 mm, and that of the horizontal ED length ranged from 0.44 to 0.46 mm. These differences in ED measurement were not significantly different from 0.5 mm (all P > 0.06). However, pairs of examiners differed in vertical ED length measurements by >0.5 mm in 44% to 52% of EDs and by >1.0 mm in 13% to 17% of EDs. Pairs of examiners differed in horizontal ED length measurements by >0.5 mm in 31% to 40% of EDs and by >1.0 mm in 10% to 15% of EDs. The intraclass correlation coefficient was 0.85 (95% confidence interval, 0.77-0.91) for vertical and 0.84 (95% confidence interval, 0.74-0.90) for horizontal ED measurements. Cornea specialists showed good reliability in the measured EDs; however, depending on the threshold for clinical significance, a nontrivial percentage of cases have high interexaminer clinical variability.


[Pharmacological treatment of epilepsies: progress during the last 10 years].
The pharmacological treatment of epilepsies represents an important domain of neurological therapeutical strategies and is not only important for specialized epilepsy centres, but for all hospital -and office-based neurologists. The choice of the right anticonvulsive drug is not always simple since the development of new drugs during the last 10 years offers a broad range of possibilities. Therefore, pharmacological treatment has become more sophisticated than it was with relatively few anticonvulsive drugs before. The new drugs are better tolerated and have less side effects, but more knowledge is required about the specific aspects of each drug. Pharmacology, pharmacocinetics, indication, dosage and side effects of the substances will be summarized and discussed.


Long-term proton pump inhibitors use and its association with premalignant gastric lesions: a systematic review and meta-analysis.
Background: Long-term maintenance therapy with proton pump inhibitors (PPIs) is a common treatment strategy for acid-related gastrointestinal diseases. However, concerns have been raised about the potential increased risk of gastric cancer and related precancerous lesions with long-term PPI use. This systematic review and meta-analysis aimed to evaluate this potential risk. Methods: We searched PubMed, Embase, and the Cochrane Central Register of Controlled Trials for randomised controlled trials published before 1 March 2023, with no language restrictions. The primary endpoint was the occurrence and progression of gastric mucosal atrophy, intestinal metaplasia, Enterochromaffin-like (ECL) cell hyperplasia, gastric polyps, and gastric cancer during the trial and follow-up. Data were analysed using a random effects model. Results: Of the 4,868 identified studies, 10 met the inclusion criteria and were included in our analysis, comprising 27,283 participants. Compared with other treatments, PPI maintenance therapy for more than 6 months was associated with an increased risk of ECL cell hyperplasia (OR 3.01; 95% CI 1.29 to 7.04; p = 0.01). However, no significant increase was found in the risk of gastric mucosal atrophy (OR 1.01; 95% CI 0.55 to 1.85; p = 0.97), intestinal metaplasia (OR 1.14; 95% CI 0.49 to 2.68; p = 0.76), gastric polyps (OR 1.13; 95% CI 0.68 to 1.89; p = 0.64), or gastric cancer (OR 1.06; 95% CI 0.79 to 1.43; p = 0.71). Conclusion: This systematic review and meta-analysis does not support an increased risk of gastric cancer or related precancerous lesions with long-term PPI maintenance therapy. However, long-term PPI use should be monitored for potential complications such as ECL cell hyperplasia. Further studies are needed to confirm these findings and evaluate the safety of PPI maintenance therapy for acid-related gastrointestinal diseases. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/, Identifier: PROSPERO (CRD42022379692).


Controlled Substance Reconciliation Accuracy Improvement Using Near Real-Time Drug Transaction Capture from Automated Dispensing Cabinets.
Accurate accounting of controlled drug transactions by inpatient hospital pharmacies is a requirement in the United States under the Controlled Substances Act. At many hospitals, manual distribution of controlled substances from pharmacies is being replaced by automated dispensing cabinets (ADCs) at the point of care. Despite the promise of improved accountability, a high prevalence (15%) of controlled substance discrepancies between ADC records and anesthesia information management systems (AIMS) has been published, with a similar incidence (15.8%; 95% confidence interval [CI], 15.3% to 16.2%) noted at our institution. Most reconciliation errors are clerical. In this study, we describe a method to capture drug transactions in near real-time from our ADCs, compare them with documentation in our AIMS, and evaluate subsequent improvement in reconciliation accuracy. ADC-controlled substance transactions are transmitted to a hospital interface server, parsed, reformatted, and sent to a software script written in Perl. The script extracts the data and writes them to a SQL Server database. Concurrently, controlled drug totals for each patient having care are documented in the AIMS and compared with the balance of the ADC transactions (i.e., vending, transferring, wasting, and returning drug). Every minute, a reconciliation report is available to anesthesia providers over the hospital Intranet from AIMS workstations. The report lists all patients, the current provider, the balance of ADC transactions, the totals from the AIMS, the difference, and whether the case is still ongoing or had concluded. Accuracy and latency of the ADC transaction capture process were assessed via simulation and by comparison with pharmacy database records, maintained by the vendor on a central server located remotely from the hospital network. For assessment of reconciliation accuracy over time, data were collected from our AIMS from January 2012 to June 2013 (Baseline), July 2013 to April 2014 (Next Day Reports), and May 2014 to September 2015 (Near Real-Time Reports) and reconciled against pharmacy records from the central pharmacy database maintained by the vendor. Control chart (batch means) methods were used between successive epochs to determine if improvement had taken place. During simulation, 100% of 10,000 messages, transmitted at a rate of 1295 per minute, were accurately captured and inserted into the database. Latency (transmission time to local database insertion time) was 46.3 ± 0.44 milliseconds (SEM). During acceptance testing, only 1 of 1384 transactions analyzed had a difference between the near real-time process and what was in the central database; this was for a "John Doe" patient whose name had been changed subsequent to data capture. Once a transaction was entered at the ADC workstation, 84.9% (n = 18 bins; 95% CI, 78.4% to 91.3%) of these transactions were available in the database on the AIMS server within 2 minutes. Within 5 minutes, 98.2% (n = 18 bins; 95% CI, 97.2% to 99.3%) were available. Among 145,642 transactions present in the central pharmacy database, only 24 were missing from the local database table (mean = 0.018%; 95% CI, 0.002% to 0.034%). Implementation of near real-time reporting improved the controlled substance reconciliation error rate compared to the previous Next Day Reports epoch, from 8.8% to 5.2% (difference = -3.6%; 95% CI, -4.3% to -2.8%; P < 10). Errors were distributed among staff, with 50% of discrepancies accounted for by 12.4% of providers and 80% accounted for by 28.5% of providers executing transactions during the Near Real-Time Reports epoch. The near real-time system for the capture of transactional data flowing over the hospital network was highly accurate, reliable, and exhibited acceptable latency. This methodology can be used to implement similar data capture for transactions from their drug ADCs. Reconciliation accuracy improved significantly as a result of implementation. Our approach may be of particular utility at facilities with limited pharmacy resources to audit anesthesia records for controlled substance administration and reconcile them against dispensing records.


Comparing live lecture, internet-based & computer-based instruction: A randomized controlled trial.
Comparing computer and internet based instruction with traditional giving lecture would provide enough evidence to identify best teaching practice. In this study, we compared lecture, interactive internet based and computer based learning regarding medical students' knowledge acquisition and satisfaction in teaching pathophysiology of hematology and oncology. Eighty four medical students were randomized into three groups and an identical faculty member conducted the instructions through the above mentioned methods. Students' knowledge was assessed one week before and immediately after the interventions by pre and posttest. Students' satisfaction was assessed using a validated 5-point Likert scale. The results showed that students' satisfaction was significantly higher in interactive internet based group than other ones (p=0.05). There were a significant increase between pre and posttest scores in all groups (p=0.000). We used ANCOVA to compare score changes in the study groups, with posttest scores as the dependent factor and pretest scores as covariate and knowledge acquisition was significantly higher in interactive internet based group than other two groups (p=0.026). The study showed that although interactive internet based instruction is a difficult and time consuming method, it is recommended to integrate this method to medical curricula.


Adverse cutaneous reactions to ticlopidine in patients with coronary stents.
A prospective study was carried out to assess the occurrence and character of adverse cutaneous reactions in patients receiving ticlopidine hydrochloride to prevent subacute thrombosis after having undergone placement of coronary stents. During a 1-year period such patients were requested to report any adverse cutaneous reactions, and those with skin reactions were referred for dermatological evaluation. Among the 136 patients who underwent stent placement by one of the authors, 20 were referred for dermatological evaluation. Of these, 16 (11.8%) fit the case definition of ticlopidine-associated cutaneous reactions. In the first 8 consecutive patients ticlopidine was withdrawn (in 2 of these a rechallenge test was later performed); in the next 8 patients ticlopidine was not discontinued before completion of the intended 4-week period of treatment. Patients remained under weekly follow-up and underwent a weekly blood count. Skin biopsies were obtained in 5 patients with different types of eruptions. The skin reactions appeared from 2 to 21 days after commencement of ticlopidine (mean, 10 days), lasting from 2 to 30 days (mean, 5 days). Only 3 patients had other adverse effects: neutropenia in 1 and abdominal pain and nausea in 2. The most common presentations were urticaria, pruritus, and maculopapular eruption. In 3 patients there were previously unreported reactions: fixed drug eruption, erythromelalgia-like eruption, and erythema multiforme-like eruption. Of note was the rapid clearing of the skin eruption in most cases even when the drug was not withdrawn. It was concluded that adverse cutaneous reactions are relatively common in association with ticlopidine treatment but that serious reactions are rare and the disappearance of the signs and symptoms is rapid, suggesting that discontinuation of the drug is not usually imperative.


Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells.
The acidic environment of the lysosomal lumen provides an optimal milieu for the acid hydrolases and is also essential for fusion/fission of endo-lysosomal compartments and sorting of cargo. Evidence suggests that maintaining lysosomal acidity is essential to avoid disease. In this chapter, we describe a protocol for analyzing the lysosomal pH in cultured cells using the fluorescent probe fluorescein isothiocyanate (FITC)-dextran together with a dual-emission ratiometric technique suitable for flow cytometry. Fluorescence-labeled dextran is endocytosed and accumulated in the lysosomal compartment. FITC shows a pH-dependent variation in fluorescence when analyzed at maximum emission wavelength and no variation when analyzing at the isosbestic point, thereby the ratio can be used to determine the lysosomal pH. A standard curve is obtained by equilibrating intralysosomal pH with extracellular pH using the ionophore nigericin. The protocol also includes information regarding procedures to induce lysosomal alkalinization and lysosomal membrane permeabilization.


Free catecholamine excretion in the urine in normal infants and in those with marasmus or kwashiorkor.
Free catecholamine (epinephrine + norepinephrine) excretions of normal male infants 2.1-3.2, 4.5-10.6, and 12.5-18.5 months of age, respectively, and of infants and children with marasmus or marasmic kwashiorkor were measured on three consecutive days after admission and after partial rehabilitation. In normal infants, particularly the older ones, the first day's excretion was higher than that of the next two days, probably in response to the stress of the procedures. There was an increase with age, but on a surface area basis, the differences were not significant, the three age groups excreting 15.1 plus or minus 10.2, 23.8 plus or minus 20.9, and 24.7 plus or minus 14.3 mug/m-2/day, respectively. Excretions of marasmic infants on admission were not significantly different from those of the control children. Higher mean values were due to elevated excretions of infants with severe infection. After partial rehabilitation, excretions were similar to those of control subjects. In children with marasmic kwashiorkor excretions were no different from those of control infants, except in severely infected children. Admission values revealed more day-to-day variation than recovery values or than those of normal and marasmic infants, in whom subject to subject variation was more marked.


Physicochemical characterization of a PEGylated liposomal drug formulation using capillary electrophoresis.
In this work, the applicability of using CE to perform a physicochemical characterization of a PEGylated liposomal drug formulation of the anti-cancer agent oxaliplatin was examined. Characterization of the liposomal drug formulation using CE instrumentation encompassed: determination of the electrophoretic mobilities, size determination by Taylor dispersion analysis and interaction studies. Electrophoretic mobilities determined by CE were compared with the results obtained by laser Doppler electrophoresis, which were found to be subject to larger variation. Average hydrodynamic diameters of the liposome preparations, as determined by Taylor dispersion analysis, were in the range of 61-84 nm and were compared with the results obtained by dynamic light scattering. Interactions between oxaliplatin (and paracetamol) and the PEGylated liposome were non-detectable by CE frontal analysis as well as by liposome electrokinetic chromatography. In contrast, for the more lipophilic compound propranolol, apparent liposome-aqueous phase distribution coefficients (D(lip) ) were successfully determined by both electrokinetic chromatography (log D(lip) =2.10) and by CE frontal analysis (log D(lip) =2.14). It is envisioned that CE and capillary-based techniques, including Taylor dispersion analysis, will be useful tools for the characterization of nanoparticulate (e.g. liposomal) drug formulations.


An Evaluation of Commonly Used External Ventricular Drain Securement Methods in a Porcine Model: Recommendations to Improve Practice.
External ventricular drain (EVD) dislodgement is common and leads to significant morbidity and mortality. Many securement techniques to prevent this are described. There are, however, no objective studies comparing them. This study aimed to determine the most secure method of securing an EVD. A survey was distributed through the British Neurosurgical Trainee Research Collaborative to determine common EVD securement methods and select techniques for testing. Securement methods were tested in a pig cadaver model. Peak pull force before EVD failure was measured. Failure was defined as catheter displacement 1 cm from the insertion site, catheter fracture, or suture fracture. Twenty-three neurosurgical units responded. Five basic EVD securement methods were in common use. These were tested in isolation and in combination so that in total 15 common methods were tested. The most secure method was a triple construct, consisting of an anchoring suture, sutures around a coil of the catheter, and either a soft plastic flange (25.85 N, 95% confidence interval 24.95 N-26.75 N) or a hard plastic flange (29.05 N, 95% confidence interval 25.69 N-32.41 N). Of the individual methods, single anchoring sutures, soft flanges, VentriFix, and staples were found to be the least secure, whereas multiple sutures and hard flanges were the most secure. An anchoring suture followed by a coil of the catheter and finally a flange is the most secure method for securing EVDs. This simple technique can withstand up to 8.2 times the force of a single anchoring suture, is easily used, and decreases the likelihood of EVD dislodgement and associated complications.


Risk Factors for Failure of Cast Treatment in Displaced Pediatric Tibial Shaft Fractures.
As there is a current increasing tendency to treat displaced tibial shaft fractures in adolescents surgically, it has become more important to predict failure of cast treatment for these patients. In the past, redisplacement of pediatric tibial shaft fractures has been reported at rates of 20% to 40%. Although the efficacy of the three-point index (TPI), gap index, and cast index has been demonstrated for upper extremity fractures in children, to date no index has been shown to accurately predict redisplacement for pediatric tibial shaft fractures. The aim of this study was to determine the predictive factors for redisplacement in pediatric tibial shaft fractures. In all, 157 displaced pediatric tibial shaft fractures were evaluated retrospectively. Patient age, initial and postreduction fracture angulation, shortening and translation, quality of reduction, obliquity of fracture, associated fibular fractures, and 3 indices (TPI, cast index, and gap index) were analyzed. Receiver operating characteristic analysis was performed to determine the cutoff points and logistic regression was used to show the risk factors of redisplacement. There were 53 female and 104 male patients with a mean age of 9.1 (5 to 15 y) and 45 patients developed redisplacement during the follow-up. Mean TPI and gap index and initial and postreduction fracture translation were higher in patients with redisplacement, while TPI>0.855 and postreduction translation >18% were the only independent risk factors for fracture redisplacement. No differences were observed regarding associated fibular fracture, quality of reduction, initial/postreduction angulation, and shortening. The TPI>0.855 and postreduction translation >18% are independent risk factors for redisplacement of tibial shaft fractures in children. Although the gap index can be useful, the cast index is not an appropriate tool for these fractures.


Latent Lyme neuroborreliosis: presence of Borrelia burgdorferi in the cerebrospinal fluid without concurrent inflammatory signs.
Borrelia burgdorferi, the etiologic agent of Lyme borreliosis, was isolated from the CSF of a patient with elevated serum IgG antibody titers against B burgdorferi and a history of multiple tick bites. The absence of concurrent inflammatory signs of CSF as well as intrathecal antibody production indicates a phase of latent Lyme neuroborreliosis in which no tissue infection or reaction has yet occurred. Bilateral tinnitus was the only clinical symptom in this patient. The persistence of the bilateral tinnitus after antibiotic therapy did not support a causal relationship between this symptom and the borrelial infection.


Noninvasive assessment of liver fibrosis using ultrasound-based shear wave measurement and comparison to magnetic resonance elastography.
Magnetic resonance elastography (MRE) has excellent performance in detecting liver fibrosis and is becoming an alternative to liver biopsy in clinical practice. Ultrasound techniques based on measuring the propagation speed of the shear waves induced by acoustic radiation force also have shown promising results for liver fibrosis staging. The objective of this study was to compare ultrasound-based shear wave measurement to MRE. In this study, 50 patients (28 female and 22 male; age range, 19-81 years) undergoing liver MRE examinations were studied with an ultrasound scanner modified with shear wave measurement functionality. For each patient, 27 shear wave speed measurements were obtained at various locations in the liver parenchyma away from major vessels. The median shear wave speed from all measurements was used to calculate a representative shear modulus (μ) for each patient. Magnetic resonance elastographic data processing was done by a single analyst blinded to the ultrasound measurement results. Ultrasound and MRE measurements were correlated (r = 0.86; P < .001). Receiver operating characteristic (ROC) analysis was applied to the ultrasound measurement results with the MRE diagnosis as the "ground truth." The area under the ROC curve for separating patients with minimum fibrosis (defined as μ(MRE) ≤2.9 kPa) was 0.89 (95% confidence interval, 0.77-0.95), and the area under the ROC curve for separating patients with advanced fibrosis (defined as μ(MRE) ≥5.0 kPa) was 0.96 (95% confidence interval, 0.87-0.99). Results indicate that the ultrasound-based shear wave measurement correlates with MRE and is a promising method for liver fibrosis staging.


How Do Sport Parents Engage in Autonomy-Supportive Parenting in the Family Home Setting? A Theoretically Informed Qualitative Analysis.
The purpose of this study was to identify and examine how sport parents engage in autonomy-supportive parenting in the family home setting. A total of 44 parents and children from 19 families were initially interviewed. Data from these families were profiled to identify seven families that adopted a highly autonomy-supportive parenting style. The seven families' data were then examined using a theoretically focused qualitative analysis using the three dimensions of autonomy-supportive parenting. Sport parents engaged in autonomy support (vs. control) through flexible conversations and supporting decision making. The themes of boundary setting and establishing expectations based on values were indicative of structure. The authors found high levels of involvement across contexts. These findings depict the nature and types of social interactions in the family home that created an autonomy-supportive emotional climate, which often extended to sport, providing a foundation for future theoretical development and applied research in sport.


Endovascular versus surgical revascularization in proximal subclavian artery obstruction.
The aim of this paper was to compare the outcomes of endovascular versus surgical treatment in patients with symptomatic proximal subclavian artery obstruction through a retrospective clinical study. Treatment of symptomatic subclavian artery obstruction can be performed with percutaneous transluminal angioplasty or open surgical reconstruction. Comparative studies are scarce. Technical success, patency and complication rates of 47 endovascular reconstructions in 46 patients were retrospectively compared with those of 19 open surgical reconstructions in 17 patients performed between 1996 and 2012. An additional series of 51 surgical reconstructions performed in the same institution between 1976 and 1993 served as a reference. The technical success rate was 79% for endovascular and 100% for open surgical reconstructions (P<0.05). Primary patency was 72% and 89% at 1 year or 54% and 55% at 5 years for the endovascular and open surgical groups, respectively (log rank 0.210, P=0.65). Assisted primary patency was 77% and 100% at 1 year or 67% and 67% at 5 years, respectively (log rank 0.528, P=0.47). There was no mortality and major complications were infrequent, occurring equally in both groups (P=0.22). Although with its less invasive character endovascular treatment has gained preference over surgical treatment of proximal subclavian obstruction in many cases, extrathoracic surgical reconstruction can be performed with a higher technical success rate, similar patency and a comparable number of complications.


Housewife or working mum--each to her own? The relevance of societal factors in the association between social roles and alcohol use among mothers in 16 industrialized countries.
To investigate whether differences in gender-income equity at country level explain national differences in the links between alcohol use, and the combination of motherhood and paid labour. Cross-sectional data in 16 established market economies participating in the Gender, Alcohol and Culture: An International Study (GenACIS) study. Population surveys. A total of 12,454 mothers (aged 25-49 years). Alcohol use was assessed as the quantity per drinking day. Paid labour, having a partner, gender-income ratio at country level and the interaction between individual and country characteristics were regressed on alcohol consumed per drinking day using multi-level modelling. Mothers with a partner who were in paid labour reported consuming more alcohol on drinking days than partnered housewives. In countries with high gender-income equity, mothers with a partner who were in paid labour drank less alcohol per occasion, while alcohol use was higher among working partnered mothers living in countries with lower income equity. In countries which facilitate working mothers, daily alcohol use decreases as female social roles increase; in contrast, in countries where there are fewer incentives for mothers to remain in work, the protective effect of being a working mother (with partner) on alcohol use is weaker. These data suggest that a country's investment in measures to improve the compatibility of motherhood and paid labour may reduce women's alcohol use.


Laparoscopic cholecystectomy in situs inversus totalis: a case report.
Laparoscopic cholecystectomy is one of the commonest surgical procedures carried out in the world today. Occasionally patients present with undiagnosed situs inversus and acute cholecystitis. We discuss one such case and outline how the diagnosis was made and the pitfalls encountered during surgery and how they were overcome. A 32 year old female presented to our department with epigastric pain radiating through to the back. A diagnosis of acute cholecystitis in a patient with situs inversus totalis was made following clinical examination and radiological investigation. Laparoscopic cholecystectomy was subsequently performed and the patient made an uneventful recovery. Situs inversus presenting with acute cholecystitis is very rare. The surgeon must appreciate that care should be taken to set up the operating theatre in the mirror image of the normal set-up for cholecystectomy, and that right handed surgeons must modify their technique to adapt to the mirror image anatomy.


[Cellular viability and immune response in homologous cryopreserved cardiac valves].
Long-term durability of homograft valves is related to cellular viability and immunological compatibility. In this paper, we carry out a critical update of the relationship between cellular viability and immunology of cryopreserved homograft valves. In the first section, we review the theoretical background of the superior durability of cryopreserved valves with high cellular viability, with respect to the "fresh" antibiotic-stored valves, with lower viability rates. Afterwards, a brief review of the principal factors influencing the homograft valve viability rate during harvesting, preparation, sterilization and cryopreservation phases, is performed. In the second paragraph, we analyze the problem of cellular viability in cryopreserved valves, both for matrix cells--fibroblast--and for endothelium. In particular, we report the current methods of quantifications of viability and more recent biological researches. Correlations between biological and clinical data are also discussed. Finally, the immunological issue and its relationships with cellular viability is analyzed, with particular regard to experimental evidence and clinical implications. Moreover, we have described the recent hypothesis on the influence of cryopreservation on the antigenicity of tissues, and the laboratory researches on the long-term antibody response in humans after homograft valve implantation.


Outcomes of polyethylene liner cementation into a fixed metal acetabular shell with minimum follow-up of 7 years.
Cementation of a polyethylene liner into the well-fixed shell is a convenient option for revision total hip arthroplasty. We retrospectively reviewed 45 patients who had liner cementation to investigate the risk factors which gave rise to major complications and reoperation. Patients were observed for a minimum of 7 years (range 7.8-14 years). Relevant risk factors (age, BMI, surgical approach, previous cup size and position, types of coated surface) were assessed with Cox regression analysis. The mean Harris Hip Score was improved from 62.5 (range 57-68) preoperatively to 87.1 (range 70-97). A total of 7 hips (15.5%) had acetabular component loosening that was treated with reoperation. Prevalence of acetabular component loosening was statistically significantly higher in hydroxyapatite-coated group (5 of 13) than in the Ti-coated group (2 of 32, p = 0.015). All recurrent dislocations occurred in patients treated with a posterior approach. Diameter of the previous metal shell of below 54 mm showed a lower 10-year survival rate than those greater than 54 mm in diameter. PE liner cementation in stable metal cup is a useful alternative option for carefully selected patients. Pre-existing HA-coated cups as well as small sized cups were indicative of poor outcomes.


Overexpression of BCL-x protein in primary breast cancer is associated with high tumor grade and nodal metastases.
Dysregulation of genes that control apoptosis can contribute to tumor progression and increased drug resistance. The BCL2 gene and its family member BCL-x as well as the TP53 genes regulate apoptosis and have been shown to have a direct effect on the sensitivity of cancer cells to radiation and chemotherapeutic agents. The expression of BCL-x, a BCL2-related protein that is a potent inhibitor of apoptosis, was investigated by immunohistochemical and immunoblot methods in 43 primary untreated breast carcinomas, in conjunction with BCL2 and TP53. BCL-x protein was overexpressed in 18 of 42 (43%) invasive breast cancers when compared with adjacent normal breast epithelium. Western blot analysis of eight primary breast cancers and five breast cancer cell lines indicated that BCL-xL was the predominant BCL-x protein expressed. Overexpression of BCL-x protein in these tumors was associated with higher tumor grade and increased number of positive nodes. In contrast, BCL2 protein was overexpressed in 19 of 42 tumors (45%) and was strongly correlated with estrogen receptor positivity, lower tumor grade, smaller tumor size, and lower stage. TP53 protein immunostaining was detected in 12 of 40 tumors (29%) and was inversely correlated with BCL2 expression and ER positivity. There was no correlation between the level of BCL-x protein expression and age, tumor size, ER status, and TP53 status. At a median follow-up time of 216 weeks, there was a trend toward decreased overall survival in patients with tumors overexpressing BCL-x. These findings suggest that expression of BCL-x protein is increased in a significant fraction of invasive breast cancers. In contrast to BCL2 expression, up-regulation of BCL-x protein may be a marker of tumor progression. Additional data including larger numbers of patients, more uniform treatments, and longer follow-up are needed to define the prognostic significance of overexpression of BCL-x during breast cancer progression.


Adapting qualitative research strategies to technology savvy adolescents.
To adapt research strategies involving adolescents in a grounded theory qualitative research study by conducting email rather than face-to-face interviews. Adolescent culture relies heavily on text-based communication and teens prefer interactions mediated through technology. Traditional qualitative research strategies need to be rethought when working with adolescents. Adapting interviewing strategies to electronic environments is timely and relevant for researching adolescents. Twenty three adolescents (aged 16-21) were interviewed by email. A letter of invitation was distributed. Potential participants emailed the researcher to convey interest in participating. If the inclusion criteria were met, email interviews were initiated. Participants controlled the interviews through their rate of response to interview questions. A grounded theory methodology was employed. Initial contact with participants reiterated confidentiality and the ability to withdraw from the study at any time. Interviews began with the collection of demographic information and a broad opening based on a semi-structured interview guide. All data were permissible, including text, photos, music, videos or outside media, for example YouTube. The participant was allowed to give direction to the interview after initial questions were posed. Email interviews continued until saturation was reached in the data. Participants were enthusiastic about email interviewing. Attrition did not occur. Email interviewing gave participants more control over the research, decreased power differentials between the adolescent and researcher, allowed the study to be adapted to cultural, linguistic and developmental needs, and maintained confidentiality. As participants said that email communication was slow and they preferred instant messaging, replication in faster-paced media is recommended. Repetition in face-to-face settings is warranted to evaluate how technology may have influenced the findings. Implications for practice/research Adolescents' use of the internet and their preference for textbased communication makes a compelling support for modifying traditional face-to-face qualitative investigations to reflect these changing contextual conditions.


On formation of melanosomes in a cultured melanoma line.
Melanosomes are present in abundant numbers in HPM-73 melanoma cells maintained in surum-containing cultures. However, withholding of serum for 3 days caused severe retardation of development of these organelles. Addition of chondroitin-4-sulphate (0.4 mg/ml) to serum-containing culture media for 3 days resulted in greater numbers of premelanosomes and melanosomes relative to mitochondria. The same amount of chrondroitin-4-sulphate (0.4 mg/ml) added to serum-free cultures also resulted in an increased ratio of premelanosomes and melanosomes to mitochondria but, within the 3-day period many of these organelles did not attain the pattern of maturity evident in melanoma cells reared in serum-supplemented media. By exploitation of slowed rates of development of melanosomes in cultures (especially, but not exclusively, in serum-free samples) it was possible to examine various early developmental forms of these organelles. It was found that in this cell line mitochondria appear to be involved in the process of melanosome formation.


Modular synthesis of oligoacetylacetones via site-selective silylation of acetylacetone derivatives.
Oligoacetylacetones consisting of 3,3-disubstituted pentane-2,4-diones were synthesized through a terminal silylation and oxidative coupling protocol. Highly selective formation of mono-enol silyl ethers of 3,3-disubstituted acetylacetones was achieved using 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as a base. Subsequent silver(i) oxide mediated coupling reactions provided tetraketones. Unique substituent dependence was found for the terminal-selective silylation of tetraketones. Finally, octaketones (tetramers of acetylacetone derivatives) with three types of monomer sequences were prepared in their discrete forms. Single crystal X-ray analysis revealed that the solid-state conformations of oligoketone chains were predominantly governed by the ketone sequence rather than substituents. However, differences in the packing structures induced by alkyl substituents led to significant differences in melting points for the structural isomers of octaketones.


MHC class II and non-MHC class II genes differentially influence humoral immunity to Bacillus anthracis lethal factor and protective antigen.
Anthrax Lethal Toxin consists of Protective Antigen (PA) and Lethal Factor (LF), and current vaccination strategies focus on eliciting antibodies to PA. In human vaccination, the response to PA can vary greatly, and the response is often directed toward non-neutralizing epitopes. Variable vaccine responses have been shown to be due in part to genetic differences in individuals, with both MHC class II and other genes playing roles. Here, we investigated the relative contribution of MHC class II versus non-MHC class II genes in the humoral response to PA and LF immunization using three immunized strains of inbred mice: A/J (H-2k at the MHC class II locus), B6 (H-2b), and B6.H2k (H-2k). IgG antibody titers to LF were controlled primarily by the MHC class II locus, whereas IgG titers to PA were strongly influenced by the non-MHC class II genetic background. Conversely, the humoral fine specificity of reactivity to LF appeared to be controlled primarily through non-MHC class II genes, while the specificity of reactivity to PA was more dependent on MHC class II. Common epitopes, reactive in all strains, occurred in both LF and PA responses. These results demonstrate that MHC class II differentially influences humoral immune responses to LF and PA.


Effects of endurance training on oxidative capacity and structural composition of human arm and leg muscles.
Six healthy subjects performed endurance training of the same duration with legs and arms consecutively. Performance and muscle structure were measured before and after training in lower and upper limbs. Training induced similar increases in maximal oxygen consumption (6 +/- 1 vs. 7 +/- 2 mL min-1 kg-1: legs vs. arms, P > 0.05) and mitochondrial volume in leg and arm muscles (42 +/- 12 vs. 31 +/- 11%: legs vs. arms, P > 0.05). The gain in mitochondrial volume after training was achieved solely by increasing the fraction of mitochondria (+40 +/- 11%, P < 0.05) in the same muscle volume (+2 +/- 2%, P > 0.05) in the legs. In contrast, increased muscle volume (+14 +/- 3%, P < 0.05), in addition to a tendency for an increase in mitochondrial fraction (+16 +/- 11%, P > 0.05), occurred in the arms after training. Thus, similar improvements in muscle oxidative capacity in upper and lower limbs were brought about by different mechanisms. It is suggested that due to infrequent use and a lack of load-bearing function, arm muscle volume is underdeveloped in untrained, sedentary or detrained/injured subjects and that the mode of endurance training used in this study is sufficient to enlarge arm muscle volume as well as aerobic capacity.


BET 2: Is prehospital focused abdominal ultrasound useful during triage at mass casualty incidents?
A short-cut review was carried out to determined whether the addition of prehospital focused abdominal ultrasound to triage protocols might reduce time to necessary surgery and reduce overall mortality. Thirty-five papers were found using the reported searches, of which three presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results and study weaknesses of those best papers are shown in table 2. It is concluded that although the feasibility of prehospital ultrasound in mass casualty incidents has been demonstrated, there is, as yet, no clear evidence of benefit as part of a triage protocol.


Antibiotics in the treatment of peri-implantitis.
To review and discuss current literature on the use of systemically administered or locally delivered antibiotics in the treatment of peri-implantitis. A literature search was conducted using MEDLINE through the PubMed database of the US National Library of Medicine using studies up to 2011. Studies on the microbiology of peri-implantitis lesions were hand selected. Two studies on the use of systemically administered antibiotics in the treatment of peri-implantitis were identified. Both studies involved a case series without controls. Five studies on locally delivered antibiotics were found. In all cases, local antibiotics were used in conjunction with mechanical debridement and chemical disinfection with antimicrobial agents such as chlorhexidine digluconate or hydrogen peroxide. The additional effects of local antibiotics were noted in all studies but were in general moderate. This may in part be due to the selection of patients with advanced deep pockets and advanced bone loss. The current available scientific information on the use of locally or systemically administered antibiotics is insufficient to allow any firm specific recommendations for the use of these drugs. Local application of minocycline or doxycycline as an adjunct to mechanical debridement and irrigation with an antimicrobial agent may be effective in moderately deep lesions. Surgical access by full-thickness flap surgery in deeper lesions is probably necessary to halt the progression of bone loss. No sound scientific basis is available for the use of systemic antibiotics. There is an urgent need for randomised clinical trials on the use of antibiotics in the treatment of peri-implantitis. Proper periodontal infection control in dentate patients before implants are installed and frequent supportive implant care represent effective measures to reduce the risk of future infections and their complications around oral implants.


Single cell multi-omics reveal intra-cell-line heterogeneity across human cancer cell lines.
Human cancer cell lines have long served as tools for cancer research and drug discovery, but the presence and the source of intra-cell-line heterogeneity remain elusive. Here, we perform single-cell RNA-sequencing and ATAC-sequencing on 42 and 39 human cell lines, respectively, to illustrate both transcriptomic and epigenetic heterogeneity within individual cell lines. Our data reveal that transcriptomic heterogeneity is frequently observed in cancer cell lines of different tissue origins, often driven by multiple common transcriptional programs. Copy number variation, as well as epigenetic variation and extrachromosomal DNA distribution all contribute to the detected intra-cell-line heterogeneity. Using hypoxia treatment as an example, we demonstrate that transcriptomic heterogeneity could be reshaped by environmental stress. Overall, our study performs single-cell multi-omics of commonly used human cancer cell lines and offers mechanistic insights into the intra-cell-line heterogeneity and its dynamics, which would serve as an important resource for future cancer cell line-based studies.


Suicide and Suicide Risk in Adolescents.
Suicide is the second leading cause of death for 10- to 24-year-olds in the United States and is a global public health issue, with a recent declaration of a National State of Emergency in Children's Mental Health by the American Academy of Pediatrics, American Academy of Child and Adolescent Psychiatry, and Children's Hospital Association. This clinical report is an update to the previous American Academy of Pediatrics clinical report, "Suicide and Suicide Attempts in Adolescents." Because pediatricians and pediatric health care providers are at the front line of care for adolescents amid a child and adolescent mental health crisis, and because of the chronic and severe shortage of mental health specialists, it is important that pediatric health care providers become facile with recognizing risk factors associated with suicidality and at-risk populations, screening and further assessment of suicidality as indicated, and evidence-based interventions for patients with suicidal ideation and associated behaviors. Suicide risk can be mitigated by appropriate screening, bolstering of protective factors, indicated treatment, community resources, and referrals to mental health providers when available.


Neutrophil Extracellular Traps (NETs) Take the Central Stage in Driving Autoimmune Responses.
Following fifteen years of research, neutrophil extracellular traps (NETs) are widely reported in a large range of inflammatory infectious and non-infectious diseases. Cumulating evidences from in vitro, in vivo and clinical diagnostics suggest that NETs may play a crucial role in inflammation and autoimmunity in a variety of autoimmune diseases, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV). Most likely, NETs contribute to breaking self-tolerance in autoimmune diseases in several ways. During this review, we discuss the current knowledge on how NETs could drive autoimmune responses. NETs can break self-tolerance by being a source of autoantigens for autoantibodies found in autoimmune diseases, such as anti-citrullinated protein antibodies (ACPAs) in RA, anti-dsDNA in SLE and anti-myeloperoxidase and anti-protein 3 in AAV. Moreover, NET components could accelerate the inflammatory response by mediating complement activation, acting as danger-associated molecular patterns (DAMPs) and inflammasome activators, for example. NETs also can activate other immune cells, such as B cells, antigen-presenting cells and T cells. Additionally, impaired clearance of NETs in autoimmune diseases prolongs the presence of active NETs and their components and, in this way, accelerate immune responses. NETs have not only been implicated as drivers of inflammation, but also are linked to resolution of inflammation. Therefore, NETs may be central regulators of inflammation and autoimmunity, serve as biomarkers, as well as promising targets for future therapeutics of inflammatory autoimmune diseases.


Ten easy steps to conducting transparent, reproducible meta-analyses for infant researchers.
Meta-analyses provide researchers with an overview of the body of evidence in a topic, with quantified estimates of effect sizes and the role of moderators, and weighting studies according to their precision. We provide a guide for conducting a transparent and reproducible meta-analysis in the field of developmental psychology within the framework of the MetaLab platform, in 10 steps: (1) Choose a topic for your meta-analysis, (2) Formulate your research question and specify inclusion criteria, (3) Preregister and document all stages of your meta-analysis, (4) Conduct the literature search, (5) Collect and screen records, (6) Extract data from eligible studies, (7) Read the data into analysis software and compute effect sizes, (8) Visualize your data, (9) Create meta-analytic models to assess the strength of the effect and investigate possible moderators, (10) Write up and promote your meta-analysis. Meta-analyses can inform future studies, through power calculations, by identifying robust methods and exposing research gaps. By adding a new meta-analysis to MetaLab, datasets across multiple topics of developmental psychology can be synthesized, and the dataset can be maintained as a living, community-augmented meta-analysis to which researchers add new data, allowing for a cumulative approach to evidence synthesis.


Acoustic neuroma surgery: an eclectic approach with emphasis on preservation of hearing. The New York University-Bellevue experience.
During the past 10 years, 157 patients have been operated on at the New York University-Bellevue Medical Center for acoustic neuromas and other cerebellopontine angle tumors. We describe our diagnostic protocol with joint neurosurgical evaluation leading to either translabyrinthine (otology only) or suboccipital-transmeatal (combined otology-neurosurgery) surgery. The decision is made on the basis of tumor size, level of hearing, age, and health of the patient. Using these criteria, 105 suboccipital and 59 translabyrinthine operations were performed with eight patients having had two-stage operations. In 12 of 29 patients, hearing was successfully preserved. Of 18 patients with good hearing and extracanalicular tumors less than 2 cm, hearing was preserved in 11. We describe the surgical technique used in this suboccipital-transmeatal operation and present illustrative cases in detail.


[Regeneration of the ciliary beat of human ciliated cells].
The influence of an isotonic, alkaline saline solution (diluted "Emser Sole" or brine from the spa of Bad Ems) on the ciliary beat of isolated cultured human ciliated cells of the upper respiratory tract was investigated. The ciliary beat was observed via an inverted phase contrast microscope (Zeiss Axiomat IDPC) and measured microphotometrically under physiological conditions and after the damaging influence of 1% propanal solution. Under physiological conditions the saline solution had a positive, although statistically not significant influence on the frequency of the ciliary beat. After damage of the cultivated cells by 1% propanal solution, the saline solution had a significant better influence on the regeneration of the cultured cells than a physiological sodium chloride solution. It is concluded that diluted brine from Bad Ems has a positive effect on the ciliary beat of the respiratory epithelium and accelerates its regeneration after damage by viral and bacterial infections, surgery or inhaled noxae.


Cell-free translation of mitochondrial nicotinamide nucleotide transhydrogenase.
Mammalian nicotinamide nucleotide transhydrogenase is translated as a 5000 daltons larger molecular weight precursor in a cell-free system programmed with rat liver polysomes. The mature rat liver enzyme had the same molecular weight as the purified beef heart enzyme, 115 000 daltons. The precursor was not processed in vitro by liver mitochondria or by a rat liver mitochondrial matrix fraction, nor did it appear to bind to mitochondria. In contrast, pre-FeS protein of the cytochrome bc1 complex was processed in the same samples by both mitochondria and matrix, suggesting an important difference in the processing mechanisms or in the efficiency of processing of the two precursors.


[Experimental study of the photokilling effect of a new photosensitizer, aluminum phthalocyanine, on human hepatoma cell line].
The photosensitizing effect of sulfonated aluminum phthalocyanine (ALSPC), a new photosensitizer, on human hepatoma cells (SMMC-7721) was studied in vitro. It was found that ALSPC was not cytotoxic even at a concentration of 100 micrograms/ml. In the photosensitizing experiments with different irradiation wavebands, the red waveband had the most marked killing effect, which coincided with the peak absorption band of ALSPC. As the best penetration waveband for human body tissues is just located in this red region, ALSPC has a potential value in clinical application. The photosensitizing inactivation of cells was also estimated by 3H-TDR radioisotope labelling method, confirming that ALSPC had photokilling ability. ALSPC had almost the same photokilling activity on cancer cells as HPD.


Allosensitization does not alter post-transplant outcomes in pediatric patients bridged to transplant with a ventricular assist device.
Patients supported with a VAD are at increased risk for sensitization. We aimed to determine risk factors for sensitization as well as the impact of sensitization on post-transplant outcomes. The UNOS database (January 2004-June 2014) was used to identify patients (≤18 yrs) supported with a durable VAD. Rates and degree of sensitization in the VAD cohort were calculated. Post-transplant survival was determined comparing outcomes of sensitized vs. non-sensitized patients. There were 3097 patients included in the study; 19% (n = 579) were bridged with a VAD. Of these, 41.8% were sensitized vs. 29.9% of the patients who were not bridged with a VAD (p < 0.001). VAD was an independent predictor of sensitization (OR 2.05 [1.63-2.57]; p < 0.001). There was no difference in sensitization based on device type (continuous vs. pulsatile flow, p = 0.990). Post-transplant survival rates between the sensitized and non-sensitized VAD patients were not different, including patients with a PRA >50% and VAD patients with a positive DSC (p = 0.280 and 0.160, respectively). In conclusion, pediatric VAD patients are more likely to be sensitized, but there was no difference in sensitization based on device type. In addition, sensitization does not appear to impact outcomes.


Responses of human gingival and periodontal fibroblasts to a low-zinc environment.
Morphology, motility, proliferation rate and markers of oxidative stress in primary human gingival fibroblasts (GF) and periodontal ligamental fibroblasts (PDL-F) grown in zinc-deficient cultivation medium (ZDM), were studied over a 5-week culture period. A low-zinc environment effectively reduced the total, as well as the free, intracellular zinc content in both cell types, over the course of the experiment. Decreased intracellular zinc content resulted in altered cellular morphology, reduced motility, and rearrangement of actin and tubulin in the cytoskeleton. In addition, fibroblasts with low zinc content exhibited decreased proliferation, accompanied by changes in cell cycle distribution, expression of specific biochemical markers, increased oxidative stress and the activation of caspase-3. Supplementation of ZDM with exogenous zinc prevented the loss of intracellular zinc, while also restoring the morphology, cell proliferation and mitogenic signalling of the cultured cells. Moreover, such supplemented cells were protected against oxidative stress and cell death. Of the two primary cell cultures examined, GF were more sensitive to decreased intracellular zinc content, when compared to PDL-F. The results obtained suggest that the human primary cell cultures can be useful for the longer-term evaluation of the effects of nutritional factors originating from the environment.


Comparison of i.m. ketorolac trometamol and morphine sulphate for pain relief after cholecystectomy.
I.m. ketorolac trometamol 30 mg was compared with morphine sulphate 10 mg after cholecystectomy in a double-blind, multiple dose, randomized study of 100 patients. Assessments of pain were made immediately after operation (day 1), and the next morning (day 2). Pain intensity (verbal response score and visual analogue scale) was recorded before injection and then over a 6-h period. Pain relief was assessed also. The effect of ketorolac on operative blood loss and platelet function was examined. Time to commencing oral intake and the duration of administration of i.v. fluids were recorded. Adverse events were noted. Ketorolac produced significantly less analgesia than morphine on day 1, but on day 2 the two drugs produced a similar effect. Blood loss was not increased by ketorolac, although platelet function was impaired. Repeated i.m. administration of ketorolac did not produce any serious adverse effects.


Short-term alcohol administration alters KiSS-1 gene expression in the reproductive hypothalamus of prepubertal female rats.
Kisspeptins bind to the G-protein-coupled receptor (GPR54) to activate hypothalamic luteinizing hormone releasing hormone (LHRH) secretion at the time of puberty. Alcohol (ALC) causes depressed prepubertal LHRH release, resulting in depressed luteinizing hormone (LH) secretion and delayed puberty. Because KiSS-1 and GPR54 are important to the onset of puberty, we assessed the effects of chronic ALC administration on basal expression of these puberty-related genes within the reproductive hypothalamus, as well as hormones and transduction signaling pathways contributing to their activity. Immature female rats were fed a liquid diet containing ALC for 6 days beginning when 27 days old. Controls received either companion isocaloric liquid diet or rat chow and water. Animals were decapitated on day 33, in the late juvenile stage of development. Blood was collected for the assessment of serum hormone levels. Brain tissues containing the anteroventral periventricular (AVPV) and arcuate (ARC) nuclei were obtained for assessing expression of specific puberty-related genes and proteins. KiSS-1 mRNA levels in the AVPV and ARC nuclei were suppressed (p < 0.001) in the ALC-treated rats. GPR54 gene and protein expressions were both modestly increased (p < 0.05) in AVPV nucleus, but not in ARC nucleus. Alcohol exposure also resulted in suppressed serum levels of insulin-like growth factor-1 (IGF-1), LH, and estradiol (E(2)). As IGF-1, in the presence of E(2), can induce expression of the KiSS-1 gene, we assessed the potential for ALC to alter IGF-1 signaling in the reproductive hypothalamus. IGF-1 receptor gene and protein expressions were not altered. However, protein expression of phosphorylated Akt, a transduction signal used by IGF-1, was suppressed in the AVPV (p < 0.05) and ARC (p < 0.01) nuclei. Alcohol causes suppressed KiSS-1 gene expression in the reproductive hypothalamus; hence, contributing to this drug's ability to cause suppressed LHRH secretion and disruption of the pubertal process. We suggest that this action, at least in part, is through altered IGF-1 signaling.


Occurrence of Cryptosporidium andersoni in Brazilian cattle.
Feces were collected from 68 dairy cattle, 1 to 12 mo of age, on 12 farms in the municipality of Campos dos Goytacazes, Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium sp. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) of the 18S rRNA. Four positive samples (4.54%) were sequenced and identified as Cryptosporidium andersoni. This species represents a risk for Brazilian cattle because infection can affect cattle productivity. Moreover, C. andersoni is considered a zoonotic species.