A mechanistic model of Ca regulation of thin filaments in cardiac muscle.
We present a model of Ca-regulated thin filaments in cardiac muscle where tropomyosin is treated as a continuous elastic chain confined in the closed position on the actin helix by electrostatic forces. The main distinction from previous works is that the intrinsic stress-free helical shape of the tropomyosin chain was taken into account explicitly. This results in the appearance of a new, to our knowledge, tension-like term in the energy functional and the equilibrium equation. The competitive binding of calcium and the mobile segment of troponin-I to troponin-C were described by a simple kinetic scheme. The values of dimensionless model parameters were estimated from published data. A stochastic Monte Carlo simulation of calcium curves has been performed and its results were compared to published data. The model explains the high cooperativity of calcium response of the regulated thin filaments even in the absence of myosin heads. The binding of myosin heads to actin increases the calcium sensitivity while not affecting its cooperativity significantly. When the presence of calcium-insensitive troponin-C was simulated in the model, both calcium sensitivity and cooperativity decreased. All these features were previously observed experimentally.


Application of flow cytometry for rapid bacterial enumeration and cells physiological state detection to predict acidification capacity of natural whey starters.
Natural whey starter cultures are undefined microbial communities mainly consisting of thermophilic lactic acid bacteria (LAB). The technological pressure that shapes the natural whey starter community before and during the back-slopping procedure can impact the amount and viability of the different thermophilic LAB. Traditional culture-dependent analytical methods are useful for evaluating natural whey cultures based on plate enumeration with various culture media and are commonly used as self-control procedures in dairy items. These methods have high variability and require days to obtain results. As the dairy industry has been searching for a solution to this problem for a long time, researchers must explore alternative methods for the technological evaluation of natural whey and assessment of the health status of the thermophilic acidifying bacteria community in the cheesemaking process. The flow cytometry approach has been considered an alternative to classical methods in this work sector. This study compared bacterial enumeration by plate counting and flow cytometry on natural whey samples. Flow cytometry results showed positive agreement with a tendency to overestimate, linearity, and correlation with plate counting. Other parameters have also been introduced for evaluating a natural whey starter, measuring the physiological state of the cells. Specifically, cell-wall damage and metabolic activity were also evaluated, allowing us to quantify the number of cells even in sub-optimal physiological conditions.


Retinal involvement in acute thrombotic thrombocytopenic purpura: a case report.
Thrombotic thrombocytopenic purpura (TTP) is a life-threatening hematological condition associated with deficiency in ADAMTS13. Ocular manifestations associated with TTP are uncommon. Here we report a case where a 44 year old female patient with a history of symptomatic anemia and cotton wool appearance in retina during ophthalmological examination and subsequently, she was diagnosed to have TTP. The proper management with Therapeutic Plasma Exchange (TPE) and IV methylprednisolone improved the condition of the patient markedly. It concludes that even though the presence of cotton wool appearance in retina is a nonspecific sign it needs to be properly evaluated as there can be underlying serious illnesses as in our patient. Cotton wool spots can be an early sign of underlying retinal compromise and it should be identified early.


Disposition of sulfonamides in food-producing animals: pharmacokinetics of sulfathiazole in sheep.
Plasma and urine data on sheep following administration of sulfathiazole as single intravenous and oral doses were examined. A one-compartment open model was developed to describe the pharmacokinetics of sulfathiazole in sheep. The drug was rapidly eliminated, primarily by renal excretion of unchanged sulfathiazole and metabolism to acetyl sulfathiazole, with a biological half-life of 1.3 hours. Sulfathiazole was absorbed slowly (half-life, 18 hours) and relatively completely (73%) after oral administration in solution.


Identification of a repetitive genomic sequence that is distributed among a select group of mycoplasmas.
Homologous DNA sequences common to different Mycoplasma species were identified and isolated using interspecies DNA hybridization. Specific homologies were found in three species of porcine mycoplasmas that were not detected in species from other animal hosts. Restriction fragments bearing these common sequences were isolated from genomic libraries of M. hyorhinis and M. hyopneumoniae, and DNA sequence analysis confirmed the sequence conservation predicted from hybridization studies. Multiple homology signals were detected by probing M. hyorhinis genomic digests with the isolated restriction fragments, suggesting that these common sequences are present in many copies in that species. The repetition of this sequence is, however, more limited in the other porcine mycoplasma chromosomes.


Organizational culture as a mediator of credible leadership influence on work engagement: empirical studies in private hospitals in East Java, Indonesia.
The success of health services in hospitals depends on the work engagement of medical and non-medical personnel in providing quality services for patients. Engaged employees will be more proactive, take the initiative to collaborate with others, and are committed to achieving high-quality performance. A leader's ability in an organization is a major factor in building a work ethic in the organization, instilling values, norms, and ethics through organizational culture into employee work behavior, such as work engagement. Credibility leadership is the practice of leading an organization through a positive culture change. Credible leadership can increase work engagement through an organizational culture emphasized by education and organizational leaders. The study aimed to analyze the influence of leadership credibility and organizational culture on work engagement by sampling medical and non-medical personnel at two private hospitals in Madiun City. The hypothesis was tested using structural equation modeling (SEM) and path analysis. The results of hypothesis testing show that: (1) credible leadership has a significant positive influence on organizational culture; (2) organizational culture has a significant positive influence on work engagement; (3) organizational culture is the perfect mediator of credible leadership influences on work engagement. This study's results confirm that leaders' role in shaping a positive organizational culture through good credible leadership practices, while organizational culture can increase employee work engagement.


Calcium influx through annexin V ion channels into large unilamellar vesicles measured with fura-2.
A new assay for the calcium channel activity of annexin V was developed. The calcium-sensitive fluorescence indicator, fura-2, was incorporated into large unilamellar vesicles (LUV). After establishing a calcium gradient across the liposomal membranes, native or mutated annexin V was added. The resulting calcium influx into the LUV detected through the fluorescence changes of fura-2 was used as a qualitative test for the electrophysiological properties of annexin V.


Metatranscriptomics of cheese microbial communities: Efficiency of RNA extraction from various cheese types and of mRNA enrichment.
Microbial communities from cheeses contribute to the development of typical organoleptic properties. Metatranscriptomic analyses can be used to provide a global picture of the functioning of these communities. Our objective was to evaluate the efficiency of RNA extraction from various cheese types and to evaluate mRNA enrichment procedures for metatranscriptomic analyses. For the 32 tested cheese brands, corresponding to five cheese types, the extraction yield varied from 1 μg to 363 μg RNA per gram of cheese and, overall, the yield was lower for fresh cheeses and for the core of pressed cooked cheeses than for the other cheese types. Pressed cooked cheeses also had a lower RNA integrity than the other cheese types. For total RNA extracts from four cheeses, approximately 99% of the sequencing reads corresponded to ribosomal RNA, and mRNA enrichment by RiboPOOL and FastSelect kits decreased this percentage to a range of 75 to 97% and of 53 to 76%, respectively. Comparison of RNA libraries after mRNA enrichment with libraries of undepleted total RNA showed that the FastSelect mRNA enrichment had a lower impact on the gene expression profiles of five target cheese species than the riboPOOL kit and the oligo (dT) selection method. The procedures that we describe in the present study may be useful for metatranscriptomic analysis of various cheese types.


Single local injection of recombinant fibroblast growth factor-2 stimulates healing of segmental bone defects in rabbits.
The effects of a single local injection of recombinant human fibroblast growth factor-2 on the healing of segmental bone defects were evaluated in rabbits. One month after the external fixator originally designed for this experiment was installed in the tibia of the rabbit, a 3-mm bone defect was created by an osteotomy in the middle of the tibia and 0, 50, 100, 200, or 400 microg of fibroblast growth factor-2 in 100 microl of saline solution was injected into the defect. Injection of the growth factor increased the volume and mineral content of newly made bone at the defect in a dose-dependent manner with significant effects at concentrations of 100 microg or greater. These significant effects were observed at 5 weeks and later. One hundred micrograms of the growth factor increased the volume and mineral content of newly made bone by 95 and 36%, respectively, at 5 weeks. These results indicate that a single local injection of fibroblast growth factor-2 stimulates the healing of segmental defects. We speculate that such an injection could be clinically useful for the healing of fractures even when the fracture gap is rather large.


Visual versus non-visual selection of shell colour in an Israeli freshwater snail.
The Israeli freshwater snail Theodoxus jordani exhibits high variability both in shell colour and shell pattern.In most localities in Israel, more than 80% of the snails are black regardless of the colour of their background. Within Lake Kinneret, however, there is a significant association between the relative frequency of dark patterned shells and the distribution of a black (basalt) background.Predators of jordani include the fishes Barbus longiceps, Blennius fluviatilis, and the crab Potamon potamon. In Israel, the former two are limited in their distribution to the Kinneret. Thus, the distribution of cryptic variants of the snail is positively correlated to the distribution of some of its predators.Beyond the Kinneret, shells are black probably as a result of selection by the injurious effects of ultraviolet radiation. Within the Kinneret, snails can manage without an anti-radiation shield probably because the penetration of UV light into the water is considerably reduced, due to the daily summer storms and to the seasonal bloom of unicellular algae.Apparently, then, outside the Kinneret selective forces for crypsis are partly relaxed and their effects overidden by selective pressures for solar radiation devices; within the Kinnertt, selective pressures for anti-radiation devices are partly reduced and their effects overridden by selective pressures for crypsis.


Sensitivity and specificity of using pelvic ultrasonographic parameters combined with basal gonadotropin levels to diagnose central precocious puberty in Thai girls.
The criterion standard gonadotropin-releasing hormone (GnRH) stimulation tests to diagnose central precocious puberty (CPP) are time-consuming, inconvenient, and expensive. To determine predictive cut-off values codetermined by ultrasonographic parameters and basal gonadotropin levels in girls with premature sexual development and compare them results of criterion standard tests in a study of diagnostic accuracy. Retrospective review of hormonal investigations and ultrasonographic uterine and ovarian parameters in a consecutive sample of girls at a single center, tertiary care hospital in Bangkok, Thailand. We separated data from 68 girls (age range 2-12 years) into 2 groups based on their response to a GnRH analogue agonist stimulation test. A "prepubertal response" group included girls with premature thelarche and thelarche variants (n = 18, 6.37 ± 1.77 years) and a "pubertal response" group, including girls with CPP (n = 50, 8.46 ± 1.46 years); excluding patients with pathological causes (n = 0). The basal level of luteinizing hormone (LH) had the largest area under receiver operating characteristic curves (AUC) of 0.84; 95% confidence interval [CI] 0.74-0.93) compared with basal levels of follicle stimulating hormone (AUC 0.77; 95% CI 0.64-0.90) or estradiol (0.70; 95% CI 0.56-0.85). An optimal cut-off of 0.25 IU/L LH was related to a pubertal response to GnRH analogue agonist stimulation tests with 75.0% sensitivity, 88.9% specificity, 94.7% positive predictive value (PPV), and 57.1% negative predictive value. Uterine and ovarian cut-off volumes of 3.5 cm3 and 1.5 cm3 were related to a pubertal response with 88.6% and 76.2% PPV, respectively. A uterine width cut-off of 1.7 cm combined with a basal LH cut-off of 0.25 IU/L increased specificity and PPV to 100%. Combining uterine and ovarian ultrasonographic parameters with basal gonadotropin levels, especially uterine width and basal LH level, appears useful for diagnosis of CPP.


Interventions to increase seasonal influenza vaccine coverage in healthcare workers: A systematic review and meta-regression analysis.
Influenza vaccination is recommended for healthcare workers (HCWs), but coverage is often low. We reviewed studies evaluating interventions to increase seasonal influenza vaccination coverage in HCWs, including a meta-regression analysis to quantify the effect of each component. Fourty-six eligible studies were identified. Domains conferring a high risk of bias were identified in most studies. Mandatory vaccination was the most effective intervention component (Risk Ratio of being unvaccinated [RRunvacc] = 0.18, 95% CI: 0.08-0.45), followed by "soft" mandates such as declination statements (RRunvacc = 0.64, 95% CI: 0.45-0.92), increased awareness (RRunvacc = 0.83, 95% CI: 0.71-0.97) and increased access (RRunvacc = 0.88, 95% CI: 0.78-1.00). For incentives the difference was not significant, while for education no effect was observed. Heterogeneity was substantial (τ(2) = 0.083). These results indicate that effective alternatives to mandatory HCWs influenza vaccination do exist, and need to be further explored in future studies.


Apoptotic response of malignant rhabdoid tumor cells.
BACKGROUND: Malignant rhabdoid tumors (MRTs) are extremely aggressive and resist current radio- and chemotherapic treatments. To gain insight into the dysfunctions of MRT cells, the apoptotic response of a model cell line, MON, was analyzed after exposure to several genotoxic and non-genotoxic agents employed separately or in association. RESULTS: Fluorescence microscopy of chromatin morphology and electrophoretic analysis of internucleosomal DNA fragmentation revealed that MON cells were, comparatively to HeLa cells, resistant to apoptosis after treatment with etoposide, cisplatin (CisPt) or X-rays, but underwent some degree of apoptosis after ultraviolet (UV) C irradiation. Concomitant treatment of MON cells with X-rays or vinblastine and the phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin resulted in synergistic induction of apoptosis. Western blot analysis showed that the p53 protein was upregulated in MON cells after exposure to all the different agents tested, singly or in combination. In treated cells, the p53 downstream effectors p21WAF1/CIP1, Mdm2 and Bax were induced with some inconsistency with regard to the accumulation of p53. Poly ADP-ribose polymerase (PARP) cleavage, indicative of ongoing apoptosis, occurred in UVC-irradiated cells and, especially, in cells treated with combinations of X-rays or vinblastine with wortmannin. However, there was moderate or no PARP cleavage in cells treated with CisPt, X-rays, vinblastine or wortmannin singly or with the combinations X-rays plus CisPt or vinblastine and CisPt plus vinblastine or wortmannin. The synergistic effect on the induction of apoptosis exerted by some agent combinations corresponded with synergy in respect of MON cell growth inhibition. CONCLUSION: These results suggest abnormalities in the p53 pathway and apoptosis control in MRT cells. The Ras/PI3-K/AKT signaling pathway might also be deregulated in these cells by generating an excess of survival factors. These dysfunctions might contribute to the resistance of MRTs to current antineoplastic treatments and could warrant consideration in the search of new therapeutic approaches.


Entrainment of the circadian rhythm of wheel-running activity is phase shifted by ablation of the intergeniculate leaflet.
The phase angle of entrainment of golden hamster wheel-running activity was determined before and after bilateral destruction of the intergeniculate leaflet (IGL). Animals were maintained under continuous dim light which rhythmically varied in intensity (10-5 lux) in the form of a sine wave with a 24 h period. The phase angle of entrainment changed significantly following IGL ablation suggesting a role for the IGL in entrainment.


The pharmacokinetics and pharmacodynamics of atovaquone and proguanil for the treatment of uncomplicated falciparum malaria in third-trimester pregnant women.
To investigate the pharmacokinetics, safety and efficacy of the recommended 3-day treatment regimen of Malarone in third-trimester pregnant women with acute uncomplicated falciparum malaria. Twenty-six pregnant women in their third trimester (gestational age: 24-34 weeks) with acute uncomplicated Plasmodium falciparum malaria who fulfilled the enrollment criteria were recruited from the antenatal clinics of Mae Sot Hospital, Tak Province, Thailand, (n = 8) and the Tropical Diseases Research Centre, Ndola, Zambia (n = 18). Patients were treated with four Malarone tablets (GlaxoSmithKline: each tablet contains 250 mg atovaquone and 100 mg proguanil) once daily for 3 consecutive days. Blood samples were taken for pharmacokinetic investigations of atovaquone, proguanil, and cycloguanil up to 288 h (day 14) after the last dose. Urine samples were collected for the evaluation of proguanil and cycloguanil 0-8, 8-16, 16-24 and 24-48 h after the last dose. Efficacy assessments included the clinical and parasitological evaluation of mothers and newborns. Adverse events were evaluated at each visit to the antenatal clinics. Malarone appeared to be effective and well tolerated when used for the treatment of falciparum malaria in pregnant women. All patients showed prompt clinical improvement and the disappearance of parasitaemia after treatment. There were no serious adverse effects or unexpected adverse effects and no stillbirths or spontaneous abortions. The plasma concentration-time profiles of atovaquone and proguanil in most cases were best characterised by the two-compartment open model with zero-order input with/without absorption lag time and first-order elimination. There were no significant differences in any of the pharmacokinetic parameters of atovaquone, proguanil or cycloguanil between patients from Thailand and Zambia. For atovaquone, a Cmax of 1.33-8.33 microg/ml was reached at 2.0-9.3 h after the last dose on day 2. V/F, CL/F and t(1/2beta) were 6.9-39.5 l/kg, 83-384 ml/h/kg, and 57.8-130.8 h, respectively. The Cmax and t(max) values for proguanil versus cycloguanil were 383-918 versus 0-129 ng/ml and 3.3-8.6 versus 3-12 h, respectively. V/F, CL/F, and t(1/2beta) values for proguanil were 10.7-34.0 l/kg, 431-1,662 ml/h/kg and 11.2-30.3 h. The CL(R-CG), t(1/2z), (CG), proguanil/cycloguanil metabolic ratios, AUC ratios for proguanil to cycloguanil (AUC(PG/CG)) were 107.2-1,001 ml/h/kg, 5-95 ml/h/kg, 7.8-20.7 h, 5-57, and 4.7-20.2, respectively. The pharmacokinetics of atovaquone and cycloguanil appeared to be influenced by the pregnancy status, resulting in an decrease in the Cmax and AUC of approximately twofold.


Changing M3G/M6G ratios and pharmacodynamics in a cancer patient during long-term morphine treatment.
A cancer patient receiving long-term oral sustained-release morphine treatment and periodically presenting with unusually high plasma M3G/M6G ratios is described. We found the patient's formation of M6G more unstable and perhaps delayed compared to the formation of M3G. There is no apparent explanation for this phenomenon and the high M3G/M6G ratios had no implications for the patient's pain experience or side effects from the morphine treatment.


Predictive score for complete occlusion of intracranial aneurysms treated by flow-diverter stents using machine learning.
Complete occlusion of an intracranial aneurysm (IA) after the deployment of a flow-diverter stent is currently unpredictable. The aim of this study was to develop a predictive occlusion score based on pretreatment clinical and angiographic criteria. Consecutive patients with ≥6 months follow-up were included from 2008 to 2019 and retrospectively analyzed. Each IA was evaluated using the Raymond-Roy occlusion classification (RROC) and dichotomized as occluded (A) or residual (B/C); 80% of patients were randomly assigned to the training sample. Feature selection and binary outcome prediction relied on logistic regression and threshold maximizing class separation selected by a CART tree algorithm. The feature selection was addressed by a genetic algorithm selected from the 30 pretreatment available variables. The study included 146 patients with 154 IAs. Feature selection yielded a combination of six variables with a good cross-validated accuracy on the test sample, a combination we labeled DIANES score (IA diameter, indication, parent artery diameter ratio, neck ratio, side-branch artery, and sex). A score of more than -6 maximized the ability to predict RROC=A with sensitivity of 87% (95% CI 79% to 95%) and specificity of 82% (95% CI 64% to 96%) in the training sample. Accuracy was 86% (95% CI 79% to 94%). In the test sample, sensitivity and specificity were 89% (95% CI 77% to 98%) and 60% (95% CI 33% to 86%), respectively. Accuracy was 81% (95% CI 69% to 91%). A score was developed as a grading scale for prediction of the final occlusion status of IAs treated with a flow-diverter stent.


Single-channel properties and pH sensitivity of two-pore domain K+ channels of the TALK family.
The two-pore K2P channel family comprises TASK, TREK, TWIK, TRESK, TALK, and THIK subfamilies, and TALK-1, TALK-2, and TASK-2 are functional members of the TALK subfamily. Here we report for the first time the single-channel properties of TALK-2 and its pHo sensitivity, and compare them to those of TALK-1 and TASK-2. In transfected COS-7 cells, the three TALK K2P channels could be identified easily by their differences in single-channel conductance and gating kinetics. The single-channel conductances of TALK-1, TALK-2, and TASK-2 in symmetrical 150 mM KCl were 21, 33, and 70 pS (-60 mV), respectively. TALK-2 was sensitive mainly to the alkaline range (pH 7-10), whereas TALK-1 and TASK-2 were sensitive to a wider pHo range (6-10). The effect of pH changes was mainly on the opening frequency. Thus, members of the TALK family expressed in native tissues may be identified based on their single-channel kinetics and pHo sensitivity.


Use of bioinformatic analyses in identifying characteristic genes and mechanisms active in the progression of idiopathic thrombocytopenic purpura in individuals with different phenotypes.
To explore the mechanism underlying the progression of newly diagnosed idiopathic thrombocytopenic purpura (ITP) to its chronic or remission state using bioinformatic methods. GSE56232 and GSE46922 gene expression profile datasets were downloaded from Gene Expression Omnibus (GEO). Differentially expressed genes were identified and characteristic genes were screened by weighted gene co-expression network analysis. These genes were used for function enrichment analysis and construction of a protein-protein interaction network. Finally, characteristic genes were verified to determine potential molecular mechanisms underlying ITP progression. We found that characteristic genes in the chronic ITP group were mainly involved in intracellular processes and ion binding, while characteristic genes in the remission ITP group were involved in intracellular processes and nuclear physiological activities. We identified a sub-network of characteristic genes, LMNA, JUN, PRKACG, SMC3, which may indicate the mechanism by which newly diagnosed ITP progresses to chronic. Although no meaningful signaling pathways were found, the expression of NR3C1, TPR, SMC4, PANBP2, CHD1, and U2SURP may affect ITP progression from newly diagnosed to remission. Our findings improve the understanding of the pathogenesis and progression of ITP, and may provide new directions for the development of treatment strategies.


Simultaneous identification of orthopoxviruses and alphaviruses by oligonucleotide macroarray with special emphasis on detection of variola and Venezuelan equine encephalitis viruses.
The development of a method in macroarray format for the identification of alphaviruses and orthopoxviruses in samples of concern in biodefense is reported. Capture oligonucleotides designed to bind generic members of the orthopox- or alphavirus families and a collection of additional oligonucleotides to bind specifically nucleic acids from five individual alphaviruses, including Venezuelan equine encephalitis, or DNA from each of four orthopoxviruses, including variola virus (VAR) were deposited onto nylon membranes. Hybridization of digoxigenin labeled PCR products to the macroarray produced results easily observable to the naked eye. Multiplex RT-PCR utilizing both orthopox- and alphavirus-generic primers yielded amplification of DNA corresponding to the expected sizes of the orthopoxvirus and alphavirus fragments, respectively. Hybridization of samples to capture oligonucleotides in the macroarray membranes identified correctly generic orthopox- or alphaviral sequences. The hybridizations correctly identified each of the three alphaviruses and two orthopoxviruses tested. We observed cross-hybridization only once (between two alphaviruses) that was less intense than the spots formed by correct hybridization. The macroarray test described below is easy to perform, inexpensive, relatively fast, uncomplicated to interpret, and its end point is read visually without the need of additional equipment. This nucleic acid hybridization assay onto nylon membranes in macroarray format can help in detecting or excluding the presence of threat viruses in environmental samples and appears promising for a variety of biodefense applications.


CT-monitored minimal ablative margin control in single-session microwave ablation of liver tumors: an effective strategy for local tumor control.
To investigate the usefulness of minimal ablative margin (MAM) control by intra-procedural contrast-enhanced CT (CECT) in microwave ablation (MWA) of liver tumors. A total of 334 consecutive liver tumors (240 hepatocellular carcinomas [HCCs] and 94 colorectal liver metastases [CRLMs]) in 172 patients treated with percutaneous MWA were retrospectively included. MAM of each tumor was assessed after expected ablation completion using intra-procedural CECT, allowing within-session additional ablation to any potentially insufficient margin. On immediate post-MWA MRI, complete ablation coverage of tumor and final MAM status were determined. The cumulative local tumor progression (LTP) rate was estimated by using the Kaplan-Meier method. To identify predictors of LTP, Cox regression analysis with a shared frailty model was performed. Intra-procedural CECT findings prompted additional ablation in 18.9% (63/334) of tumors. Final complete ablation coverage of tumor and sufficient MAM were determined by MRI to be achieved in 99.4% (332/334) and 77.5% (259/334), and their estimated 6-month, 1-year, and 2-year LTP rates were 3.2%, 7.5%, and 12.9%; and 1.0%, 2.1%, and 6.9%, respectively. Insufficient MAM on post-MWA MRI, perivascular tumor location, and tumor size (cm) were independent risk factors for LTP (hazard ratio = 14.4, 6.0, and 1.1, p < 0.001, p = 0.003, and p = 0.011, respectively), while subcapsular location and histology (HCC vs CRLM) were not. In MWA of liver tumors, intra-procedural CECT monitoring of minimal ablative margin facilitates identification of potentially suboptimal margins and guides immediate additional intra-session ablation to maximize rates of margin-sufficient ablations, the latter being a highly predictive marker for excellent long-term local tumor control. • In MWA of liver tumors, intra-procedural CECT can identify potentially suboptimal minimal ablative margin, leading to immediate additional ablation in a single treatment session. • Achieving a finally sufficient ablative margin through the MWA with intra-procedural CECT monitoring of minimal ablative margin results in excellent local tumor control.


How many distal bolts should be used in unreamed intramedullary nailing for diaphyseal tibial fractures?
Unreamed intramedullary nailing (UIMN) is an effective treatment procedure for the majority of tibial fractures, with locking constituting the technical support for the buttressing and neutralisation principles underlying intramedullary nailing. It has been claimed that the added versatility obtained from the use of more bolts in distal locking is very important. Several studies have been made concerning the optimum number of locking bolts in distal tibial fractures; however, to the best of our knowledge, no study has dealt with the question of whether two or three bolts should be used in diaphyseal fractures. In this paper, we evaluate the results of treating 86 diaphyseal tibial fractures (type 42 according to the AO classification) with Expert Tibial UIMN (Synthes™, West Chester, PA, USA) and distal blocking with either two or three bolts. Mean patient age was 35 years (21-51). We found that the consolidation time is shorter, less radiation time is needed and the material cost is lower when two bolts are used. No other differences were found regarding mean operative time, wound healing, pain at fracture site, joint function, angular deviation or rotation. For type 42 AO tibial fractures treated with Expert Tibial UIMN, distal blocking should be performed with only two bolts.


Hepatic Artery Thrombosis After Orthotopic Liver Transplant: Experience in the Last 10 Years.
Hepatic artery thrombosis (HAT) is the second cause of graft failure, after primary disfunction. It has a significant morbidity, with a retransplant and mortality rate in early hepatic artery thrombosis of 50%. The incidence of this event goes from 2% to 9% in the adult population. The objective is to assess the incidence of HAT in a third-level hospital. The study design is an observational retrospective study, collecting data of the transplant recipient from 2010 to 2020. Incidence of HAT was 5.33% (39/732). A statistical difference was found with the blood intraoperative administration (P = .002) and with the presence of anatomic abnormalities in the hepatic artery between the HAT and the non-HAT group. We did not find any statistical difference with portal thrombosis (P = .73) between the groups. HAT is a fatal complication after an orthotopic liver transplant, which can lead to graft loss and even recipient death. For these reasons, we should early identify risk factors associated with this event early and try to minimize them to avoid the devastating consequences.


Chest pain--no difference in severity between those having a myocardial infarction and chest pain from other causes.
In considering nonpleuritic chest pain, the severity of the maximal pain as indicated by the patients on a visual analogue scale does not significantly differ for myocardial infarction or chest pain of other origin. There is also no significant change with time in the severity score of the maximal pain for either group. Thus, the time course is not diagnostically useful.


The diffuse and macular atrophic dermatosis. A precocious and progressive light degenerative condition.
Two brothers with the "diffuse and macular atrophic dermatosis" making a total of 4 patients are reported. The familial occurrence, numerous large "peau citréine" plaques situated on the dorsum of the upper trunk and congenital phimosis with superficial atrophy of glans penis, represent and additional features reported. The light and electron microscopic findings of the least affected site on the arm, of a "peau citréine" and of an erythemoscleroatrophic plaque, none previously studied, have been reported. Among other items, it was also found that the elastic stain-positive globe-like structures found in an unusually large number throughout the edematous upper dermis represent more advancely transformed "Civatte bodies" containing epidermal filamentous cell material.


Specific binding sites for [3H]Ro 5-4864 in rat prostate and seminal vesicle.
1. The peripheral-type benzodiazepine receptor was characterized in rat prostate and seminal vesicle using [3H]Ro 5-4864 as radioligand. 2. The affinity of this radioligand for this receptor was higher in rat prostate (KD = 4.36 +/- 1.02 nM) than in seminal vesicle (KD = 8.45 +/- 1.34 nM). 3. The density of binding sites obtained in these two tissues was Bmax = 4164 +/- 873 fmol/mg in prostate and 5978 +/- 1022 fmol/mg in seminal vesicle. 4. The [3H]Ro 5-4864 binding was inhibited non-competitively by atractyloside and alpha,beta-methylene-ATP, suggesting a modulation by the ADP/ATP mitochondrial carrier. 5. Flutamide was able to displace bound [3H]Ro 5-4864 with an IC50 similar to protoporphyrin IX.


Detecting Medicine Safety Signals Using Prescription Sequence Symmetry Analysis of a National Prescribing Data Set.
Medicine safety signal detection methods employed by the medicine regulator in Australia (Therapeutic Goods Administration [TGA], Department of Health) rely predominantly on analysis of spontaneous adverse event (AE) reports, sponsor notifications or information shared by international agencies. The limitations of these methods and the availability of large administrative health data sets has given rise to greater interest in the use of administrative health data to support pharmacovigilance (PV). We explored whether prescription sequence symmetry analysis (PSSA) of Pharmaceutical Benefits Scheme (PBS) data can enhance signal detection by the TGA, using the AE, heart failure (HF) as a case study. We applied the PSSA method to all single-ingredient medicines dispensed under the PBS between 2012 and 2016, using furosemide initiation as a proxy for new-onset HF. A signal was considered present if the lower limit of the 95% confidence interval for the adjusted sequence ratio was > 1. We excluded medicines known to cause HF, indicated for HF treatment or indicated for diseases that may contribute to HF. Of the 654 tested medicines, 26 potential new HF signals were detected by PSSA. Five signals had additional support for the possible association provided by biological plausibility, consistency and disproportionate reporting of cases of HF to the TGA and the World Health Organization; and clinical impact. PSSA was able to identify potential signals for further evaluation. With the increasing availability of different administrative health data sources, the strengths and weaknesses of methods used to analyse these data for the purpose of regulatory PV should be evaluated.


A role of melatonin in neuroectodermal-mesodermal interactions: the hair follicle synthesizes melatonin and expresses functional melatonin receptors.
Since mammalian skin expresses the enzymatic apparatus for melatonin synthesis, it may be an extrapineal site of melatonin synthesis. However, evidence is still lacking that this is really the case in situ. Here, we demonstrate melatonin-like immunoreactivity (IR) in the outer root sheath (ORS) of mouse and human hair follicles (HFs), which corresponds to melatonin, as shown by radioimmunoassay and liquid chromatography/tandem mass spectrometry (LC/MS/MS). The melatonin concentration in organ-cultured mouse skin, mouse vibrissae follicles, and human scalp HFs far exceeds the respective melatonin serum level and is significantly increased ex vivo by stimulation with norepinephrine (NE), the key stimulus for pineal melatonin synthesis. By real-time PCR, transcripts for the melatonin membrane receptor MT2 and for the nuclear mediator of melatonin signaling, retinoid orphan receptor alpha (ROR)alpha, are detectable in murine back skin. Transcript levels for these receptors fluctuate in a hair cycle-dependent manner, and are maximal during apoptosis-driven HF regression (catagen). Melatonin may play a role in hair cycle regulation, since its receptors (MT2 and RORalpha) are expressed in murine skin in a hair cycle-dependent manner, and because it inhibits keratinocyte apoptosis and down-regulates ERalpha expression. Therefore, the HF is both, a prominent extrapineal melatonin source, and an important peripheral melatonin target tissue. Regulated intrafollicular melatonin synthesis and signaling may play a previously unrecognized role in the endogenous controls of hair growth, for example, by modulating keratinocyte apoptosis during catagen and by desensitizing the HF to estrogen signaling. As a prototypic neuroectodermal-mesodermal interaction model, the HF can be exploited for dissecting the obscure role of melatonin in such interactions in peripheral tissues.


Understanding geographical variations in health system performance: a population-based study on preventable childhood hospitalisations.
To investigate interdistrict variations in childhood ambulatory sensitive hospitalisation (ASH) over the years. Observational population-based study over 2008-2018 using the Primary Health Organisation Enrolment Collection (PHO) and the National Minimum Dataset hospital events databases. New Zealand primary and secondary care. All children aged 0-4 years enrolled in the PHO Enrolment Collection from 2008 to 2018. ASH. Only 1.4% of the variability in the risk of having childhood ASH (intracluster correlation coefficient=0.014) is explained at the level of District Health Board (DHB), with the median OR of 1.23. No consistent time trend was observed for the adjusted childhood ASH at the national level, but the DHBs demonstrated different trajectories over the years. Ethnicity (being a Pacific child) followed by deprivation demonstrated stronger relationships with childhood ASH than the geography and the health system input variables. The variation in childhood ASH is explained only minimal at the DHB level. The sociodemographic variables also only partly explained the variations. Unlike the general ASH measure, the childhood ASH used in this analysis provides insights into the acute conditions sensitive to primary care services. However, further information would be required to conclude this as the DHB-level performance variations.


Bilateral multifocal lower extremity localized soft tissue amyloidomas: case report with ultrasonographic characterization.
Amyloidosis may be acquired or hereditary and consists of multiple disease processes characterized by the abnormal deposition of extracellular fibrillary protein aggregates. The presentation of amyloidosis is varied, ranging from an isolated, focal deposition to systemic disease. While systemic involvement is common, a rare entity known as amyloidoma or tumor amyloidosis may also occur. The 75-year-old female in this case presented with slowly growing multifocal bilateral lower extremity masses and was ultimately diagnosed with localized amyloidomas in the absence of chronic illness or systemic disease. This case report includes a description of the sonographic features of localized extremity amyloidoma that, to the best of our knowledge, have not yet been described in the literature. Awareness of the sonographic features, and this unique presentation of multifocal soft tissue extremity amyloidomas will allow for this rare diagnosis to be included in a limited differential diagnosis.


Control of heat-induced cutaneous vasodilatation in relation to age.
Well matched unacclimatised older (age 55-68, 4 women, 2 men) and younger (age 19-30, 4 women, 2 men) subjects performed 75 min cycle exercise (approximately 40% VO2max) in a hot environment (37 degrees C, 60% rh). Rectal temperature (Tre), mean skin temperature (Tsk), arm blood flow (ABF, strain gauge plethysmography), and cardiac output (Q, CO2 rebreathing) were measured to examine age-related differences in heat-induced vasodilatation. Tre and Tsk rose to the same extent in each group during the exposure. There was no significant intergroup difference in sweat rate (older: 332 +/- 43 ml.m-2.h-1, younger: 435 +/- 49 ml.m-2.h-1; mean +/- SEM). However, the older subjects responded to exercise in the heat with a lower ABF response which could be attributed to a lower Q for the same exercise intensity. The slope of the ABF-Tre relationship was attenuated in the older subjects (9.3 +/- 1.3 vs 17.9 +/- 3.3 ml.100 ml-1.min-1.degrees C-1, p less than 0.05), but the Tre threshold for vasodilatation was about 37.0 degrees C for both groups. These results suggest an altered control of skin vasodilatation during exercise in the heat in older individuals. This attenuated ABF response appears to be unrelated to VO2max, and may reflect an age-related change in thermoregulatory cardiovascular function.


Nurse practitioner-patient discourse: uncovering the voice of nursing in primary care practice.
This study focused on nurse practitioner-patient conversations. The purpose was to uncover a process that might contribute to understanding the positive outcomes of nurse practitioner (NP) care. Transcripts from 24 audiotaped NP-patient conversations and three NP-researcher interviews, as well as 100 hours of field notes from observations, were analyzed using a combination of discourse and ethnographic analysis.


The influence of familiarity on memory for faces and mask wearing.
During the COVID-19 pandemic, the wearing of face masks became mandatory in public areas or at workplaces in many countries. While offering protection, the coverage of large parts of our face (nose, mouth and chin) may have consequences for face recognition. This seems especially important in the context of contact tracing which can require memory of familiar and unfamiliar contacts and whether they were wearing a mask. In this study, we tested how well participants perform at remembering faces and whether they wore a mask, and if this depends on familiarity. Our results show that: (a) Participants remembered familiar faces better than unfamiliar ones, regardless of mask wearing. (b) Participants remembered unmasked faces better than masked faces, regardless of familiarity. (c) Participants were significantly worse at remembering whether an unfamiliar face was wearing a mask or not-even if they indicated remembering the face. (d) Participants showed a bias to indicate no memory of unfamiliar faces. (e) Participants showed a bias to indicate that unfamiliar faces wore a mask, even if they did not. In sum, it was harder to remember both, the identity of unfamiliar faces and whether they wore a mask. These findings have practical relevance for contact tracing and epidemic control.


The mental health impact of the COVID-19 pandemic on Canadian critical care nurses.
Focusing on Canadian critical care nurses (CCNs), the study objectives were to examine the impact of the COVID-19 pandemic on: mental health, quality of work life, and intent to stay in their current positions. Mixed-methods study using an online cross-sectional survey and integration of closed- and open-ended survey data. Canadian CCNs working in an intensive care unit, high acuity unit, or intensive care step-down unit during the COVID-19 pandemic between May 2021 to June 2021. The survey consisted of four instruments: (1) the impact of event scale - revised, (2) the depression, anxiety, and stress scale, (3) the professional quality of life scale, and (4) intent to turnover tool, as well as one optional open-ended question. From across Canada, 425 CCNs responded. The large majority reported symptoms of post traumatic stress disorder (74%), depression (70%), anxiety (57%), and stress (61%). All (100%) reported moderate to high burnout, 87% were suffering from signs of secondary traumatic stress, and 22% intended to quit their current employment. Qualitative analysis of written comments submitted by 147 (34.5%) of the respondents depicted an immense mental health toll on CCNs that stemmed from 1) failed leadership and 2) the traumatic nature of the work environment, that led to 3) a sense of disillusionment, defeat, and an intent to leave. The mental health toll of the pandemic has been significant for Canadian CCNs and highlights the urgent need for individual supports and systems level changes.


Insurers' Negotiating Leverage and the External Effects of Medicare Part D.
By influencing the size and bargaining power of private insurers, public subsidization of private health insurance may project effects beyond the subsidized population. We test for such spillovers by analyzing how increases in insurer size resulting from the implementation of Medicare Part D affected drug prices negotiated in the non-Medicare commercial market. On average, Part D lowered prices for commercial enrollees by 3.7%. The external commercial market savings amount to $1.5 billion per year, which, if passed to consumers, approximates the internal cost-savings of newly-insured subsidized beneficiaries. If retained by insurers, it corresponds to a 5% average increase in profitability.


Value of CA 15.3 in breast cancer and comparison with CEA and TPA: a study of specificity in disease-free follow-up patients and sensitivity in patients at diagnosis of the first metastasis.
The specificity and sensitivity of a tumor marker (TM) are important in establishing its potential clinical utility for a specific type of neoplasm. CA 15.3 is a TM specific for breast cancer; it is defined by two monoclonal antibodies (DF3 and 115D8), whose specificity, in disease-free follow-up patients, and sensitivity, in patients at diagnosis of first metastasis, have been evaluated in the present study and compared with those of carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA). Serum concentrations of all three TMs were quantified in 618 individuals: 80 healthy controls, 421 patients with local breast cancer who became free of disease following locoregional treatment, and 117 patients with disseminated disease at diagnosis of metastasis. Radioimmunoassay (RIA) was the method employed, and the cut-off values obtained were 30 U/ml for CA 15.3, 5 ng/ml for CEA, and 120 U/I for TPA. The results showed CA 15.3 and CEA specificities to be analogous (95.7 and 95.5%, respectively). TPA specificity (81.9%) was lower (p < 0.001). During adjuvant therapy, CA 15.3 serum levels were seen to increase, followed by a normalization of concentration after terminating therapy. On the other hand, CA 15.3 and TPA sensitivities (64.1 and 67.5%, respectively) were greater than for CEA (44.4%, p < 0.01). It is concluded that CA 15.3 is a useful TM for breast cancer, as it offers a greater sensitivity than CEA and a higher specificity than TPA. Combining CA 15.3 and CEA fails to increase CA 15.3 sensitivity, while combining CA 15.3 with TPA increases false-positives and so likewise offers no additional benefit.


Late differentiation contributes to the apparent increase in sensory neuron number in juvenile rat.
Using both profile counts and unbiased stereological methods, estimates of neuron number in the lumbar dorsal root ganglia of the rat have been shown to increases approximately 35% during postnatal life [J. Comp. Neurol. 386 (1997) 8-15; J. Comp. Neurol. 449 (2002) 158-165]. The mechanism underlying this addition of neurons was investigated. No evidence of incorporation of (BrdU), a mitotic marker, was found. Similarly, counts of myelinated and unmyelinated axons in the sural nerve were the same in neonates and adults. These results are not consistent with the possibility that neurogenesis accounts for neuron addition. A population of neurons that stains with TuJ1, an antibody against neuronal class III beta tubulin, but not with an antibody against the phosphorylated and non-phosphorylated forms of heavy chain neurofilament protein (NF-H) was found in neonates, but not adults. These less-differentiated (type-L) neurons are not detected by either profile counts or unbiased stereology and do not transport HRP retrogradely. Maturation of this pool of incompletely differentiated neurons appears to be one mechanism whereby neuron number is augmented during postnatal life.


Neurocognitive and Information Processing Biases in Posttraumatic Stress Disorder.
We review recent research addressing neurocognitive and information processing abnormalities in posttraumatic stress disorder (PTSD), including studies informing direction of causality. We additionally consider neurocognition in the context of co-morbid mild traumatic brain injury (TBI) and psychosocial treatments for PTSD. Learning, memory, attention, inhibitory functions, and information processing biases frequently accompany PTSD, reflecting potential bi-directional relationships with PTSD. Although mild TBI is associated with increased risk of PTSD development and maintenance, TBI does not typically contribute significantly to sustained neurocognitive deficits in individuals with PTSD. Whereas better learning and memory is associated with mildly enhanced response to psychosocial interventions, such interventions may also improve neurocognitive performance and can be effectively provided to patients with TBI history. PTSD is associated with cognitive abnormalities in processing both emotionally relevant and emotionally neutral information and, although mild, may underlie some PTSD symptom expression.


A novel dual mode-of-action anti-hyperalgesic compound in rats which is neuroprotective and promotes neuroregeneration.
Chronic neuropathic pain (CNP) can result from surgery or traumatic injury, but also from peripheral neuropathies caused by diseases, viral infections, or toxic treatments. Opioids, although very effective for acute pain, do not prevent the development of CNP, and are considered as insufficient treatment. Therefore, there is high need for effective and safe non-opioid options to treat, prevent and eventually reverse CNP. A more effective approach to alleviating CNP would constitute a treatment that acts concurrently on various mechanisms involved in relieving pain symptoms and preventing or reversing chronification by enhancing both neuroprotection and neuroregeneration. We have identified and characterized GRT-X (N-[(3-fluorophenyl)-methyl]-1-(2-methoxyethyl)-4-methyl-2-oxo-(7-trifluoromethyl)-1H-quinoline-3-caboxylic acid amide), a novel drug which is able to activate both voltage-gated potassium channels of the Kv7 family and the mitochondrial translocator protein 18 kDa (TSPO). The dual mode-of-action (MoA) of GRT-X was indicated in in vitro studies and in vivo in a rat model of diabetic neuropathy. In this model, mechanical hyperalgesia was dose-dependently inhibited. After severe crush lesion of cervical spinal nerves in rats, GRT-X promoted survival, speeded up regrowth of sensory and motor neurons, and accelerated recovery of behavioral and neuronal responses to heat, cold, mechanical and electrical stimuli. These properties may reduce the likelihood of chronification of acute pain, and even potentially relieve established CNP. The absence of a conditioned place preference in rats suggests lack of abuse potential. In conclusion, GRT-X offers a promising preclinical profile with a novel dual MoA.


Localization of monoamine fluorescence in the stomatogastric nervous system of lobsters.
Explorations into the crustacean stomatogastric nervous system have focused on the stomatogastric ganglion as a paradigm of neural mechanisms and integrative circuitry. Emerging evidence implicates dopamine as a fundamental modulator of the bursting pacemaker neurons and many of the intraganglionic rhythms. The present paper has employed formaldehyde-induced fluorescence in freeze-dried specimens, a microscopic histochemical technique which demonstrates anatomical structures containing particular putative transmitters, indoleamines and catecholamines. We have localized specific, formaldehyde-induced fluorescence in the lobster stomatogastric nervous system. This fluorescence occurs throughout the stomatogastric ganglion neuropil, in 4-8 cell bodies of the bilaterally paired commissural ganglia and in discrete and continuous neurites interconnecting these 3 ganglia. Variations of the basic fluorescence technique suggest that the specific fluorescence is catecholaminergic. Evidence from related biochemical studies supports this observation and indicates that the catecholaminergic fluorescence is dopaminergic since tests for norepinephrine are negative while concomitant tests for dopamine are positive.


A comparison of the effects of pulsatile and bolus flushing methods on lipid emulsion residues that lead to bacterial growth in intravenous catheters.
The aim of this study was to compare the efficacies of pulsatile and bolus flushing methods for removing residual parenteral nutrients in common shapes of central venous (CV) and peripheral venous (PV) catheters. Straight or looped tubes filled with parenteral nutrients containing indocyanine green (ICG) were flushed with 10 mL of saline at various speeds with several pulsatile intervals. Pseudomonas aeruginosa engineered with a luciferase gene was inoculated in the flushed CV catheters. The imaging technique detected the ICG remaining and the bacterial growth in the flushed catheters. In the CV catheter, the residue was greater with pulsatile flushing with longer intervals than with bolus flushing at a relatively high flushing speed. In the PV catheter, the flushing speed with any pulsatile intervals did not affect the residue. A looped catheter showed less residue than the straight catheter regardless of the flushing condition. Bacterial growth occurred in the nutrient residue in the flushed tubes. Speed of flushing and the shape of catheters, but not pulsatile flow generation, are essential components for the efficacious removal of parenteral nutrients in the catheter. Moreover, the slight differences in remaining nutrients inside the catheter affected subsequent bacterial growth.


Levetiracetam Prevents Perforin Mediated Neuronal Injury Induced by Acute Cerebral Ischemia Reperfusion.
The purpose of this study is to explore the neuroprotection mechanism of levetiracetam (LEV) with acute focal cerebral ischemia-reperfusion (I/P) mouse. The cerebral artery I/P animal model was prepared with a middle artery cerebral occlusion method. For drug intervention, mice were intraperitoneally injected with LEV with a dose of either 15 or 150 mg/kg. Neuronal injury was evaluated by measuring the infarct area, apoptosis ratio, and observation of blood-brain barrier ultrastructure with transmission electron microscope. CD8(+) antibody and perforin antibody were used to make cross-reference screen through flow cytometry to determine a perforin-positive rate in CD8(+) T lymphocytes (PFN + %). Injection of LEV can reduce infarct area, apoptosis ratio, and blood-brain barrier damage 24 h later after acute I/P in WT mice. In vitro, perforin can lower hippocampal neuron viability. In vivo, removing perforin can relieve neuronal injury. High dose injection of LEV (150 mg/kg) can inhibit perforin secreting from CD8(+)T lymphocytes. In addition, LEV can still protect neurons with perforin knockout mice. Therefore, our results suggested that LEV may contribute to neuron protection after cerebral ischemia reperfusion. The possible mechanism may be related with perforin release. However, we cannot roll out other mechanisms.


Spectrophotometric determination of clobetasol propionate, halobetasol propionate, quinagolide hydrochloride, through charge transfer complexation.
Two spectrophotometric procedures are described for the determination of clobetasol propionate(I), halobetasol propionate(II) (corticosteroids) and quinagolide hydrochloride(III) (prolactin inhibitor). For corticosteroid drugs, the procedures are based on the formation of phenyl hydrazones of the corticosteroids which are subsequently subjected to charge transfer complexation reaction with either 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ) as pi-acceptor or with iodine as sigma-acceptor. Prolactin inhibitor was reacted directly with the previous reagents. The molar ratios of the reactants were established and the experimental conditions were studied giving maximum absorption at 588 and 290 nm with DDQ and iodine methods, respectively for the three drugs. The concentration ranges were 20-150,50-300, and 20-80 microg ml(-1) in DDQ method for (I), (II), and (III), respectively and 13-20,15-40, and 8-32 microg ml(-1) in iodine method for (I), (II) and (III), respectively.


A 12-Week Randomized Double-Blind Placebo-Controlled Clinical Trial, Evaluating the Effect of Supplementation with a Spinach Extract on Skeletal Muscle Fitness in Adults Older Than 50 Years of Age.
The aim of a 12-week randomized double-blind placebo-controlled study was to assess the effect of daily supplementation with a natural extract of Spinacia oleracea L. (4 × 500 mg capsules/day; total 2 g per day) combined with a moderate-intensity training program (1 h session/3 times a week) on skeletal muscle fitness in adults over 50 years of age. Muscle strength assessed by isokinetic and isometric dynamometry improved significantly in the experimental (n = 23) and the placebo (n = 22) groups, but the magnitude of improvement was higher in the experimental group, with between-group differences in almost all variables, including isokinetic at 60° s-1 in knee extension, peak torque (p < 0.007); total work per repetition maximum (p < 0.009); isokinetic at 180°s-1 in knee extension, peak torque (p < 0.002); total work (p < 0.007); total work per repetition maximum (p < 0.005); average power (p < 0.027); isometric in knee extension, peak torque (p < 0.005); and average peak torque (p < 0.002). Similar findings were observed for muscle quality. Changes in quality of life (SF-36) were not found, except for improvements in the role physical (p < 0.023) and role emotional (p < 0.001) domains, likely as a result of the physical training sessions. A nutritional survey did not revealed changes in dietary habits. No adverse events were recorded. In subjects over 50 years of age, moderate-intensity strength training combined with daily supplementation for 12 weeks with a natural extract of Spinacia oleracea L. improved muscle-related variables and muscle quality. Maintaining muscle health is a key component of healthy aging.


[Intra-articular proximal tibia fractures].
The incidence of proximal intra-articular tibial fractures is continuously increasing. In addition to high-energy trauma in young patients, osteoporotic fractures occur more frequently in geriatric patients. After a thorough clinical examination including X‑ray and computed tomography (CT) imaging, consolidation of the soft tissue is followed by surgical treatment to achieve the best possible anatomic reconstruction of the articular surface. Nonanatomic reduction with articular gaps >2.5 mm leads to a significantly increased risk of osteoarthritis. Selection of the surgical approach and planning of the osteosynthesis are based on the fracture morphology and the existing soft tissue damage. In addition to arthroscopically assisted percutaneous procedures, sophisticated osteosynthesis is often necessary, which requires several surgical approaches. In this context, posterior surgical approaches are becoming increasingly more important. Primary knee arthroplasty can also play a role, particularly in older patients.


[Effect of topical steroid on oral and nasal nitric oxide production in patients with allergic rhinitis].
To evaluate the effects of nasal mometasone furoate on oral and nasal nitric oxide (NO) production in patients with allergic rhinitis. Twenty-seven patients with moderate to severe symptoms of persistent allergic rhinitis were treated with mometasone furoate nasal spray (200 microg/d. qd) for 2 weeks. Nasal and oral exhaled nitric oxide concentrations, symptoms of rhinitis and quality of life were investigated before and after the treatment. There was a significant improvement in nasal exhaled nitric oxide concentrations, symptoms of rhinitis and quality of life, but not in oral exhaled nitric oxide concentrations. Subjective improvements in symptoms and quality of life did not correlate significantly with objective measurements. Our study provides subjective and objective evidence on the efficacy of intranasal mometasone furoate in improving nasal symptoms and quality of life, as well as reducing nasal inflammation.


Reliability and repeatability of the Hoffmann sign.
To determine the inter-rater, intrarater, and intrasubject reliability of the Hoffmann sign. Observational, cross-sectional study. Veterans Affairs medical center. Fifty-two consenting subjects without amputation of the first through third fingers, fixed finger contractures, relapsing remitting multiple sclerosis, or any acute central neurological illness or injury within the past 3 months requiring hospital admission were recruited from inpatients and outpatients in the Spinal Cord Injury and Physical Medicine and Rehabilitation services. The Hoffmann sign was elicited by 1 primary and 3 secondary investigators who used a standardized technique. The Hoffmann sign was considered positive if any reflexive flexion of the distal phalanx of the thumb or any of the fingers was present. In the first session, the primary and one secondary examiner performed 2 trials on both hands of each subject. Each investigator pair repeated the procedure in a second session. Cohen's κ coefficient was calculated to determine (1) inter-rater reliability, calculated per investigator pair per hand, for the first trial of a session; (2) intrarater reliability, calculated between the 2 trials of each session per investigator; and (3) intrasubject reliability, calculated per hand of each subject between the first trials of the 2 sessions. Inter-rater κ was 0.65 (188 pairs), intrarater κ was 0.89 (384 pairs), and intrasubject κ was 0.73 (178 pairs). All κ values were obtained with P < .01. The Hoffmann sign has substantial inter-rater and intrasubject reliability, in addition to outstanding intrarater reliability, when tested with the use of a standardized technique.


"AEIOU: the ABC's" of conversion from laparoscopic to open cholecystectomy.
To examine and classify the reasons for conversion and the points at which laparoscopic cholecystectomies are converted to open procedures and whether these change over time. This is a retrospective study of all patients undergoing cholecystectomy from June 1, 1990 to June 30, 1995. Reasons for conversion were classified using the "AEIOU:ABC" system developed for this study and conversion points were assigned chronologically. The "AEIOU:ABC" classification system was utilized. The most common reasons for conversion were: acute inflammation N = 61 (26.1%); adhesions N = 51 (21.8%); and organ system pathology N = 39 (16.7%). The most common conversion points were; after visualization of the peritoneal cavity but prior to dissection of the cystic structures N = 103 (44.0%); dissection of the cystic structures N = 58 (24.8%); initial laparoscopy N = 36 (15.4%). When the reasons for conversion were evaluated for changes over time there was no statistically significant change for the total group or any individual surgeon. Conversion points did not change with increasing operative experience. The "AEIOU:ABC" classification system is a simple, effective and easy to use system for classifying the myriad of reasons for conversion. The system needs to be validated prospectively not only for laparoscopic cholecystectomy but for possible application to other laparoscopic procedures.


Benson Bertheau Roe, Md, July 7, 1918-August 6, 2012.
"Ben" Roe was the eighth president of The Society of Thoracic Surgeons. He was a fourth-generation native Californian, a hall of fame athlete, a pioneer in the development of cardiac surgery, and a crusading reformer of professional standards, education, and ethics. Educated in both northern California and in Boston, his career reflected the influence of both cultures and his California heritage.


Summary measures of number needed to treat: how much clinical guidance do they provide in neuropathic pain?
Several systematic reviews of randomized controlled trials (RCTs) of drugs to treat neuropathic pain have reported summary estimates of efficacy - specifically, the number needed to treat (NNT). To examine whether summary NNTs for drugs to treat neuropathic pain provide useful guidance for clinical decision making. We examined the methods used to pool data across RCTs in systematic reviews - in particular, the extent of heterogeneity of agents (and dosages), neuropathic pain syndromes, and outcome measures. Published summary estimates of NNTs of drugs to treat neuropathic pain embody substantial heterogeneity, in that they reflect a pooling of data across agents with different mechanisms of action, across patients with different neuropathic pain syndromes, and/or across different outcome measures. Summary NNT estimates may have limited clinical relevance, due to problems of heterogeneity. The most that can be extracted from systematic reviews published to date is the identity of drugs that have demonstrated efficacy for specific types of neuropathic pain, and the strength of such evidence.


Biventricular assist device utilization for patients with morbid congestive heart failure: a justifiable strategy.
The rationale for the use of a biventricular assist device (BiVAD) for morbid congestive heart failure (MCHF) has been questioned because of historically unacceptable rates of postimplant and post-transplant mortality as well as perceived barriers to their outpatient management. All patients who received a Thoratec BiVAD from January 1990 to December 2003 at the University of Pittsburgh were studied retrospectively. There were a total of 73 patients (32% ischemic, 21% idiopathic, and 47% other) who had a BiVAD implanted. Before implantation, 100% were on > or =1 inotropic agent, and 77% had an intra-aortic balloon pump. Overall survival was 69%; 42 patients (84%) received cardiac transplantation, 5 patients (10%) were weaned, and 3 (6%) remained supported on BiVAD. If the 14 patients with postcardiotomy failure and acute myocardial infarction with shock are excluded, the overall survival improves to 75%. Five-year actuarial survival after heart transplantation was 58%. Of the 29 patients implanted before 2000, the 4-month actuarial freedom from driveline infections, bloodstream infections, and neurological events was 10%, 54%, and 48%, respectively, whereas the rates of these events for the 44 patients implanted after 2000 improved to 70%, 79%, and 80%, respectively. Since 2000, 21 (48%) patients were discharged from the hospital, of whom 38% went to an outpatient residence, 33% to a skilled nursing facility, and 29% to home. Once discharged, > or =1 readmission occurred in 45% and > or =2 readmissions in 48%. BiVAD support for MCHF has an acceptable overall mortality and survival to transplantation. Morbidity has been significantly reduced in the past 4 years, and management as an outpatient is achievable.


Electrospun glass nanofibers to strengthen polycarbonate plastic glass toward photoluminescent smart materials.
Electrospun glass nanofibers (GNFs) were used to strengthen polycarbonate (PC) to create long-persistent photoluminescent and fluorescent smart materials such as afterglow concrete and smart window. Physical integration of lanthanide-activated aluminate (LA) nanoparticles (NPs) yielded transparent GNFs@PC smart sheets. Spectral investigations utilizing photoluminescence and CIE Lab parameters were performed to confirm that the translucent appearance of GNFs@PC changed to green when exposed to UV light. This fluorescence activity was quickly reversible for the GNFs@PC hybrids with low concentrations of LANPs, which indicate fluorescence emission. Higher phosphor concentrations in GNFs@PC led to longer-lasting afterglow photoluminescence and slower reversibility. The GNFs@PC hybrids showed an emission band detected at 518 nm upon excitation at 368 nm. The morphological characteristics of LANPs and GNFs were analyzed by transmission electron microscopy (TEM), which revealed sizes of 11-26 nm and 250-300 nm, respectively. GNFs were prepared using electrospinning technology and then used as a roughening agent into PC sheets. Morphological characteristics of GNFs and GNFs@PC smart sheets were examined using energy-dispersive X-ray spectroscopy (EDXA), X-ray fluorescence (XRF) and scanning electron microscopy (SEM). The GNFs@PC smart sheets were shown to have enhanced scratch resistance in comparison to LANPs-free PC control sample. Increases in LANPs concentration enhanced both hydrophobicity and UV protection.


Pioglitazone improves left ventricular diastolic function in patients with essential hypertension.
Left ventricular (LV) hypertrophy and diastolic dysfunction, which are common cardiac consequences of hypertension, are modified by insulin resistance. The present study assessed the hypothesis that primary treatment of insulin resistance may reverse such cardiac changes in hypertensive patients. A total of 30 patients with essential hypertension were enrolled in this study. In echocardiographic examinations, LV mass index, the peak velocity ratio of early diastolic to atrial filling (E/A), and the E-wave deceleration time (DcT) were determined. Insulin sensitivity test with steady-state plasma glucose (SSPG) method, oral glucose tolerance test, and blood samplings for measurement of adiponectin and matrix metalloproteinase (MMP)-2 were also performed. Six months after treatment with pioglitazone (30 mg/day), an insulin sensitizer, these examinations were repeated. Pioglitazone significantly increased E/A and decreased DcT, without a change in LV mass index. These improvements in diastolic properties were much greater in subjects with a marked (>or==3.3 mmol/L) decrease in SSPG (n=11) than the others (n=19), although the decrease in glucose levels did not differ between the two groups. In addition, the changes in E/A and DcT were closely correlated with the decrease in SSPG. Pioglitazone treatment significantly elevated plasma adiponectin and MMP-2 levels, and the increase in MMP-2 was positively correlated with the increase in adiponectin. The present findings demonstrate that pioglitazone improves LV diastolic function without LV mass regression in hypertensive patients in proportion to the amelioration of insulin resistance. These findings suggest that increased adiponectin and MMP may be involved in the beneficial effect of pioglitazone on diastolic function.


Purification and properties of rat liver 3-hydroxy-3-methylglutaryl coenzyme A reductase.
3-Hydroxy-3-methylglutaryl coenzyme A reductase has been purified from rat liver microsomes with a recovery of approx. 25%. The enzyme was homogeneous on gel electrophoresis and enzyme activity comigrated with the single protein band. The molecular weight of the reductase determined by gel filtration on Sephadex G-200 was 200,000. SDS-polyacrylamide gel electrophoresis gave a subunit molecular weight of 52,000 +/- 2000, suggesting that the enzyme was a tetramer. The specific activities of the purified enzyme obtained from rats fed diets containing 0% or 5% cholestyramine were 11,303 and 19,584 nmol NADPH oxidized/min per mg protein, respectively. The reductase showed unique binding properties to Cibacron Blue Sepharose; the enzyme was bound to the Cibacron Blue via the binding sites for both substrates, NADPH and (S)-3-hydroxy-3-methylglutaryl coenzyme A. Antibodies prepared against purified reductase inactivated 100% of the soluble and at least 91% of the microsomal enzyme activity. Immunotitrations of solubilized enzyme obtained from normal and cholestyramine-fed rats indicated that cholestyramine feeding both increased the amount of enzyme protein and resulted in enzyme activation. Administration of increasing amounts of mevalonolactone to rats decreased the equivalence point obtained from immunotitration studies with solubilized enzyme. These data indicate that the antibody cross-reacts with the inactive enzyme formed after mevalonolactone treatment.


Benign vs malignant vertebral compression fractures with MRI: a comparison between automatic deep learning network and radiologist's assessment.
To test the diagnostic performance of a deep-learning Two-Stream Compare and Contrast Network (TSCCN) model for differentiating benign and malignant vertebral compression fractures (VCFs) based on MRI. We tested a deep-learning system in 123 benign and 86 malignant VCFs. The median sagittal T1-weighted images (T1WI), T2-weighted images with fat suppression (T2WI-FS), and a combination of both (thereafter, T1WI/T2WI-FS) were used to validate TSCCN. The receiver operator characteristic (ROC) curve was analyzed to evaluate the performance of TSCCN. The accuracy, sensitivity, and specificity of TSCCN in differentiating benign and malignant VCFs were calculated and compared with radiologists' assessments. Intraclass correlation coefficients (ICCs) were tested to find intra- and inter-observer agreement of radiologists in differentiating malignant from benign VCFs. The AUC of the ROC plots of TSCCN according to T1WI, T2WI-FS, and T1WI/T2WI-FS images were 99.2%, 91.7%, and 98.2%, respectively. The accuracy of T1W, T2WI-FS, and T1W/T2WI-FS based on TSCCN was 95.2%, 90.4%, and 96.2%, respectively, greater than that achieved by radiologists. Further, the specificity of T1W, T2WI-FS, and T1W/T2WI-FS based on TSCCN was higher at 98.4%, 94.3%, and 99.2% than that achieved by radiologists. The intra- and inter-observer agreements of radiologists were 0.79-0.85 and 0.79-0.80 for T1WI, 0.65-0.72 and 0.70-0.74 for T2WI-FS, and 0.83-0.88 and 0.83-0.84 for T1WI/T2WI-FS. The TSCCN model showed better diagnostic performance than radiologists for automatically identifying benign or malignant VCFs, and is a potentially helpful tool for future clinical application. TSCCN-assisted MRI has shown superior performance in distinguishing benign and malignant vertebral compression fractures compared to radiologists. This technology has the value to enhance diagnostic accuracy, sensitivity, and specificity. Further integration into clinical practice is required to optimize patient management. • The Two-Stream Compare and Contrast Network (TSCCN) model showed better diagnostic performance than radiologists for identifying benign vs malignant vertebral compression fractures. • The processing of TSCCN is fast and stable, better than the subjective evaluation by radiologists in diagnosing vertebral compression fractures. • The TSCCN model provides options for developing a fully automated, streamlined artificial intelligence diagnostic tool.


Protein carbonyl levels in serum and gingival crevicular fluid in patients with chronic periodontitis.
Evidence reveals the role of reactive oxygen species (ROS) in many pathologies including periodontitis. Protein carbonylation is the most widely used biomarker for oxidative damage to proteins, and reflects cellular damage induced by ROS. In this study protein carbonyl (PC) levels in serum and gingival crevicular fluid (GCF) in patients with chronic periodontitis (CP) was evaluated. Thirty-three patients with CP and 24 healthy controls were included in the study. Following the clinical measurements and samplings, total protein levels in serum and GCF were determined by Bradford method, and serum and GCF PC levels were measured by modified Levine method. PC levels in serum and GCF were significantly higher in the CP group compared to the control group (p<0.05). In all subjects, serum and GCF PC levels showed statistically significant positive correlations with all clinical parameters (p<0.05). The results suggest that both systemic and local/periodontal protein carbonylation increase in CP compared to health and that elevated levels of PCs may be a sign of oxidative damage in periodontitis and correlate well with the periodontal status.


Retrieval and the extinction of memory.
1. Memory is assessed by measuring retrieval which is often elicited by the solely presentation of the conditioned stimulus (CS). However, as known since Pavlov, presentation of the CS alone generates extinction. 2. One-trial avoidance (IA) is a much used conditioned fear paradigm in which the CS is the safe part of a training apparatus, the unconditioned stimulus (US) is a footshock and the conditioned response (CR) is to stay in the safe area. Retrieval of the memory for the step-down version of this task is measured in the absence of the US, as latency to step-down from the safe area (i.e., a platform). 3. Extinction of the IA response is installed at the moment of the first non-reinforced test session, as clearly shown by the fact that many drugs, including PKA, ERK and protein synthesis inhibitors as well as NMDA receptor antagonists, hinder extinction when infused into the hippocampus or the basolateral amygdala at the moment of the first test session but not later. 4. Some, but not all the molecular systems required for extinction are also activated by retrieval, further endorsing the hypothesis that although retrieval is necessary for the generation of extinction this last process constitutes a new learning secondary to the non-reinforced expression of the original trace.


Total chemical synthesis of histones and their analogs, assisted by native chemical ligation and palladium complexes.
Chemical synthesis of histones allows precise control of the installation of post-translational modifications via the coupling of derivatized amino acids. Shortcomings of other approaches for obtaining modified histones for epigenetic studies include heterogeneity of the obtained product and difficulties in incorporating multiple modifications on the same histone. In this protocol, unprotected peptide fragments are prepared by Fmoc solid-phase synthesis and coupled in aqueous buffers via native chemical ligation (NCL; in NCL, a peptide bond is formed between a peptide with an N-terminal Cys and another peptide having a C-terminal thioester). This task is challenging, with obstacles relating to the preparation and ligation of hydrophobic peptides, as well as the requirement for multiple purification steps due to protecting-group manipulations during the polypeptide assembly process. To address this, our approach uses an easily removable solubilizing tag for the synthesis and ligation of hydrophobic peptides, as well as a more efficient and better-yielding method to remove Cys-protecting groups that uses palladium chemistry (specifically [Pd(allyl)Cl]2 and PdCl2 complexes). The utility of this approach is demonstrated in the syntheses of ubiquitinated H2B at Lys34, phosphorylated H2A at Tyr57 and unmodified H4. Each of these analogs can be prepared in milligram quantities within ∼20-30 d.


Population Pharmacokinetic Model to Optimize Cefotaxime Dosing Regimen in Critically Ill Children.
During sepsis, optimal plasma antibiotic concentrations are mandatory. Modifications of pharmacokinetic parameters could lead to low drug concentrations and therefore, insufficient therapeutic levels. The aim of this study was to build a population pharmacokinetic model for cefotaxime and its metabolite desacetylcefotaxime in order to optimize individual dosing regimens for critically ill children. All children aged < 18 years, weighing more than 2.5 kg, and receiving intermittent cefotaxime infusions were included in this study. Cefotaxime and desacetylcefotaxime were quantified by high-performance liquid chromatography. Pharmacokinetics were described using the non-linear mixed-effect modeling software MONOLIX, and Monte Carlo simulations were used to optimize dosing regimen in order to maintain serum concentrations above the target concentration (defined at 2 mg·L-1) throughout the dosing interval. We included 49 children with a median (range) postnatal age of 23.7 (0.2-229) months, and median body weight (range) of 10.9 (2.5-68) kg. A one-compartment model with first-order elimination adequately described the data. Median (range) values for cefotaxime clearance, desacetylcefotaxime clearance, and volume of distribution were 0.97 (0.3-7.1) L·h-1, 3.2 (0.6-16.3) L·h-1, and 0.3 (0.2-0.41) L·kg-1, respectively. Body weight and postnatal age were statistically significant covariates. Cefotaxime-calculated residual concentrations were low, and no patient succeeded in attaining the target. Unlike intermittent administration, a dosing regimen of 100 mg·kg-1·day-1 administered by continuous infusion provided a probability of target attainment of 100%, regardless of age and weight. Standard intermittent cefotaxime dosing regimens in critically ill children are not adequate to reach the target. We showed that, for the same daily dose, continuous infusion was the only administration that enabled the target to be attained, for children over 1 month of age. As continuous administration is achievable in the pediatric intensive care unit, it should be considered for clinical practice. Registered at http://www.clinicaltrials.gov , NCT02539407.


Sperm cryopreservation: effects on chromatin structure.
Cryopreservation is a technique that can keep sperm alive indefinitely, enabling the conservation of male fertility. It involves the cooling of semen samples and their storage at -196°C in liquid nitrogen. At this temperature all metabolic processes are arrested. Sperm cryopreservation is of fundamental importance for patients undergoing medical or surgical treatments that could induce sterility, such as cancer patients about to undergo genotoxic chemotherapy or radiotherapy, as it offers these patients not only the hope of future fertility but also psychological support in dealing with the various stages of the treatment protocols.Despite its importance for assisted reproduction technology (ART) and its success in terms of babies born, this procedure can cause cell damage and impaired sperm function. Various studies have evaluated the impact of cryopreservation on chromatin structure, albeit with contradictory results. Some, but not all, authors found significant sperm DNA damage after cryopreservation. However, studies attempting to explain the mechanisms involved in the aetiology of cryopreservation-induced DNA damage are still limited. Some reported an increase in sperm with activated caspases after cryopreservation, while others found an increase in the percentage of oxidative DNA damage. There is still little - and contradictory - information on the mechanism of the generation of DNA fragmentation after cryopreservation. More studies are needed to establish the true importance of such damage, especially to improve the results of ART.


Urinary protein profiles after burn injury.
A study of the change of urinary protein excretion has been made in 26 patients followed from the time of admission to time of discharge from the Yorkshire Regional Burn Centre (7-200 days). Total protein, IgG and albumin (as indicators of glomerular function) and alpha 1-microglobulin, beta 2-microglobulin, retinol binding protein and N-acetyl-beta-D-glucosaminidase (as indicators of proximal tubular function) as well as the patterns on SDS polyacrylamide gel electrophoresis were examined. Patients were divided into three groups according to their urinary protein profiles. The first group was 3 patients who showed little or no evidence of proteinuria; the second group consisted of 7 patients who showed a mild and transient tubular proteinuria, all of whom recovered normal function within one week. The third group, 16 patients, developed moderate to severe proteinuria usually started as a mixed pattern of glomerular and tubular proteinuria, characterized by the excretion of a high level of total protein. After a few days, this pattern changed to a typical tubular proteinuria and the latter could last for as long as 200 days; in some of these patients a secondary increase of low molecular weight protein excretion occurred either due to a latent renal complication or induced by the administration of aminoglycosides. There is a close correlation between the intensity and patterns of the proteinuria and the severity and clinical progress of the burn.


Are there differential deficits in facial emotion recognition between paranoid and non-paranoid schizophrenia? A signal detection analysis.
This study assessed facial emotion recognition abilities in subjects with paranoid and non-paranoid schizophrenia (NPS) using signal detection theory. We explore the differential deficits in facial emotion recognition in 44 paranoid patients with schizophrenia (PS) and 30 non-paranoid patients with schizophrenia (NPS), compared to 80 healthy controls. We used morphed faces with different intensities of emotion and computed the sensitivity index (d') of each emotion. The results showed that performance differed between the schizophrenia and healthy controls groups in the recognition of both negative and positive affects. The PS group performed worse than the healthy controls group but better than the NPS group in overall performance. Performance differed between the NPS and healthy controls groups in the recognition of all basic emotions and neutral faces; between the PS and healthy controls groups in the recognition of angry faces; and between the PS and NPS groups in the recognition of happiness, anger, sadness, disgust, and neutral affects. The facial emotion recognition impairment in schizophrenia may reflect a generalized deficit rather than a negative-emotion specific deficit. The PS group performed worse than the control group, but better than the NPS group in facial expression recognition, with differential deficits between PS and NPS patients.


A high-throughput-compatible assay to measure the degradation of endogenous Huntingtin proteins.
The accumulation of disease-causing proteins is a common hallmark of many neurodegenerative disorders. Measuring the degradation of such proteins using high-throughput-compatible assays is highly desired for the identification of genetic and chemical modulators of degradation. For example, Huntington's disease (HD) is an incurable hereditary neurodegenerative disorder caused by the cytotoxicity of mutant huntingtin protein (mHTT). The high-throughput measurement of mHTT degradation is important in HD drug discovery and research. Existing methods for such purposes have limitations due to their dependence on protein tags or pan protein synthesis inhibitors. Here, we report a high-throughput-compatible pulse-chase method (CH-chase) for the measurement of endogenous tag-free huntingtin protein (HTT) degradation based on Click chemistry and Homogeneous Time Resolved Fluorescence (HTRF) technologies. The pulsed-labeled proteins were conjugated with biotin using the click reaction strain-promoted alkyne-azide cycloaddition (SPAAC), and the chase signals were calculated by measuring the reduction percentage of the HTT HTRF signals after pull-down with streptavidin beads. We validated that the signals were within the linear detection range and were HTT-specific. We successfully measured the degradation of endogenous HTT in a high-throughput-compatible format using 96-well plates. The predicted changes of HTT degradation by known modifiers were observed, which confirmed that the assay is suitable for the identification of HTT degradation modifiers. We have established the first high-throughput-compatible assay capable of measuring endogenous, tag-free HTT degradation, providing a valuable tool for HD research and drug discovery. The method could be applied to other proteins and can facilitate research on other neurodegenerative disorders and proteinopathies.


Bizarre implants for augmentation mammaplasty: long term human reaction to polyethylene strips.
Two patients are described where a mass of polyethylene strips had been used for augmentation mammaplasty. The reaction to this plastic was different in each case.


Production of pilot batches 223Ra and 227Th of medical purpose.
The pilot batches of 227ThCl4 and 223RaCl2 were separated from 227Ас. 227Th was separated by anion exchange chromatography on the sorbent BioRad AG-1x8 100-200 mesh from 8 М HNO3. After 227Th storage accumulated 223Ra was separated from 227Th using two chromatography columns: on sorbent BioRad AG-1x8 100-200 mesh from 8 М HNO3 followed by eluate evaporation, by dissolving of dry residue in 0.02 М EDTA with рН 4.5 and sorption on cation exchanger BioRad AG-50x8 200-400 mesh. The samples obtained were analyzed by gamma-, alpha- and atomic emission spectrometry methods. Long-living 227Ас content in Ra and Th was determined by a preliminary chemical separation.


Uptake and intracellular degradation of fluorescent sphingomyelin by fibroblasts from normal individuals and a patient with Niemann-Pick disease.
Sphingomyelin, labelled with a fluorescent probe, pyrene, in the fatty acyl residue was associated with fetal calf serum; approx. 80% of the sphingomyelin was found in the low- and high-density lipoproteins. This was added to the growth medium of cultured human skin fibroblasts from normal individuals and a patient with Niemann-Pick disease type A, devoid of acid sphingomyelinase activity. The fluorescent sphingomyelin was taken up by both cell types, but only the former degraded it to produce fluorescent ceramide. Differences between normal and Niemann-Pick cells in sphingomyelin content or ceramide production were observed after several hours uptake. A more pronounced difference was noted when cells were incubated for 1 day with fluorescent sphingomyelin and then for two to three days in medium devoid of this compound. Under these conditions, the fluorescence intensity of the Niemann-Pick cells remained practically constant while that of their normal counterparts was almost completely eliminated from the cells. Comparison of fluorescence intensities of these two cell types could be made directly on aqueous suspensions of whole cells or, alternatively, on their lipid extracts. For evaluation of the degradation of fluorescent sphingomyelin to ceramide within the cells, several procedures were developed for the rapid isolation of the latter compound from the total lipid extract. The results suggest that when associated with the constituents of the fetal calf serum, sphingomyelin is taken up by the cells and transported into the lysosomal compartment where it is degraded to ceramide. Use of the fluorescent derivative of sphingomyelin provided a simple and rapid procedure for following the uptake by and degradation within the cultured cells. It also permitted the establishment of differences in the rates of degradation of the fluorescent sphingomyelin by cells with a normal metabolism and others lacking sphingomyelinase (i.e., Niemann-Pick disease type A cells).


Quantitative proteomics and phosphoproteomic analyses of mouse livers after tick-borne Babesia microti infection.
Babesia microti is a tick-borne protozoan parasite that infects the red blood cells of mice, humans, and other mammals. The liver tissues of BALB/c mice infected with B. microti exhibit severe injury. To further investigate the molecular mechanisms underlying liver injury and liver self-repair after B. microti infection, data-independent acquisition (DIA) quantitative proteomics was used to analyse changes in the expression and phosphorylation of proteins in liver tissues of BALB/c mice during a B. microti infection period and a recovery period. The expression of FABP1 and ACBP, which are related to fatty acid transport in the liver, was downregulated after infection with B. microti, as was the expression of Acox1, Ehhadh and Acaa1a, which are crucial rate-limiting enzymes in the process of fatty acid β oxidation. The phosphorylation levels of AMP-activated protein kinase (AMPK) and Hormone-sensitive lipase (HSL) were also downregulated. In addition, the expression of PSMB9, CTSC, and other immune-related proteins was increased, reflecting an active immune regulation mechanism in the mice. The weights of mice infected with B. microti were significantly reduced, and the phosphorylation levels of IRS-1, c-Raf, mTOR, and other proteins related to growth and development were downregulated.


Automatic detection of three-dimensional vascular tree centerlines and bifurcations in high-resolution magnetic resonance angiography.
We sought to develop a simple and robust algorithm capable of automatically detecting centerlines and bifurcations of a three-dimensional (3D) vascular bed. After necessary preprocessing, an appropriate cost function is computed for all vessel voxels and Dijkstra's minimum-cost-path algorithm is implemented. By back tracing all the minimum-cost paths, centerlines and bifurcation are detected. The detected paths are then split into segments between adjacent nodes (bifurcations or vessel end-points) and smoothed by curve fitting. Application of the algorithm to both simulated 3D vessels and 3D magnetic resonance angiography (MRA) images of an actual intracranial arterial tree produced well-centered vessel skeletons. Quantitative assessment of the algorithm was performed. For the simulated data, the root mean square error for centerline detection is about half a voxel. For the human intracranial MRA data, the sensitivity, positive predictive value (PPV), and accuracy of bifurcation detection were calculated for different cost functions. The best case gave a sensitivity of 91.4%, a PPV of 91.4%, and an RMS error of 1.7 voxels. To the extent that imperfections are eliminated from the segmented image, the algorithm is effective and robust in automatic and accurate detection of centerlines and bifurcations. The cost function and algorithm used are demonstrated to be an improvement over similar algorithms in the literature.


The effects of general and alcohol-specific peer factors in adolescence on trajectories of alcohol abuse disorder symptoms from 21 to 33 years.
The present study examines whether general and alcohol-specific peer risk factors from age 10 to 18 are associated with longitudinal patterns of adult alcohol abuse disorder symptoms from age 21 to 33. Using growth mixture modeling, trajectory groups of alcohol abuse disorder symptoms from age 21 to 33 were identified. We then examined the relationships between the identified trajectory groups of alcohol abuse disorder symptoms and respondents' own adolescent binge drinking, a general negative peer factor, and an alcohol-specific peer factor (having drinking peers) in adolescence using pseudo-class Wald Chi-square tests, and multinomial logistic regressions. Four different trajectory groups of alcohol abuse disorder symptoms were identified: persistor group (3%), decreaser group (23%), escalator group (3%), and a no-disorder group (71%). Bivariate Wald Chi-square tests indicated that adolescent binge drinking behavior and general and alcohol-specific peer factors differentiated the adult alcohol abuse trajectory groups. Multivariate multinomial logistic regression showed that the general negative peer factors distinguished those who later persisted in alcohol abuse from those who desisted (i.e., persistor group vs. decreaser group) during young adulthood, even after adjusting for respondents' adolescent binge drinking. On the other hand, associating with drinking peers did not distinguish these trajectories. Alcohol-specific peer influences appear to influence alcohol abuse disorder symptoms in the early 20s, while general negative peer exposure in adolescence increases in importance as a risk factor for alcohol abuse disorder symptom persistence in the late 20s and the early 30s.


Humans navigate with stereo olfaction.
Human navigation relies on inputs to our paired eyes and ears. Although we also have two nasal passages, there has been little empirical indication that internostril differences yield directionality in human olfaction without involving the trigeminal system. By using optic flow that captures the pattern of apparent motion of surface elements in a visual scene, we demonstrate through formal psychophysical testing that a moderate binaral concentration disparity of a nontrigeminal odorant consistently biases recipients' perceived direction of self-motion toward the higher-concentration side, despite that they cannot verbalize which nostril smells a stronger odor. We further show that the effect depends on the internostril ratio of odor concentrations and not the numeric difference in concentration between the two nostrils. Taken together, our findings provide behavioral evidence that humans smell in stereo and subconsciously utilize stereo olfactory cues in spatial navigation.


Thrombogenicity of components of atheromatous material. An animal and in vitro model of cerebral atheroembolism.
Human atheromatous material was separated into lipid and nonlipid fractions by ether and chloroform-methyl alcohol procedures. The maximum nonlethal doses of nonlipid, lipid, and whole (unseparated) atheromatous material were 8 mg, between 15 and 30 mg, and between 50 and 60 mg when injected into the left common carotid arteries of rabbits. In vitro production of thrombi showed that the nonlipid material produced thrombi that were larger in volume, weight, and length than those produced by whole material.