Randomized discontinuation trials: utility and efficiency.
The randomized discontinuation trial (RDT) is a two-phase trial. In phase I all patients are openly treated with the medication being evaluated. In phase II, those who have responded are randomly assigned to continue the same treatment or switch to placebo. Usually, non-compliers and "adverse reactors" identified in phase I are excluded from phase II. To investigate the value of this design, we reviewed the advantages and limitations of discontinuation studies, and compared the RDT design to the classic randomized clinical trial design in terms of clinical utility and efficiency (sample size). A computer model was used to study the efficiency of the two designs under a broad range of assumptions. The RDT design is particularly useful in studying the effect of long-term, non-curative therapies, especially when the clinically important effect is relatively small, and the use of placebo should be minimized for ethical or feasibility reasons. However, its use is limited if the objective of an investigation is to estimate the magnitude of absolute treatment effects and toxic effects in the source population, or to evaluate a potentially curative treatment. Our results indicate that selecting responders prior to randomization has a very strong effect on the relative efficiency of the trial. Further improvement may be achieved by excluding non-compliers and adverse reactors. Under the assumptions tested in our model, the sample size required in phase II of an RDT was only 20-50% of that in a classic trial.


Site-directed mutagenesis of citrate synthase; the role of the active-site aspartate in the binding of acetyl-CoA but not oxaloacetate.
Asp-362, a potential key catalytic residue of Escherichia coli citrate synthase (citrate oxaloacetate-lyase [pro-3S)-CH2COO- ----acetyl-CoA), EC 188.8.131.52) has been converted to Gly-362 by oligonucleotide-directed mutagenesis. The mutant gene was completely sequenced, using a series of synthetic oligodeoxynucleotides spanning the structural gene to confirm that no additional mutations had occurred during genetic manipulation. The mutant gene was expressed in M13 bacteriophage and produced a protein which migrated in an identical manner to wild-type E. coli citrate synthase on SDS-polyacrylamide gels and which cross-reacted with E. coli citrate synthase antiserum. The mutant gene was subsequently recloned into pBR322 for large scale purification of the protein, and the resulting plasmid, pCS31, used to transform the citrate synthase deletion strain, W620. The mutant enzyme purified in an analogous manner to wild-type E. coli citrate synthase and expressed less than 2% of wild-type enzyme activity. The activity of the partial reactions catalysed by citrate synthase was similarly affected suggesting that this residual activity may be due to contaminating wild-type enzyme activity. The mutant citrate synthase retains a high-affinity NADH-binding site consistent with the protein preserving its overall structural integrity. Oxaloacetate binding to the protein is unaffected by the Asp-362 to Gly-362 mutation. Binding of the acetyl-CoA analogue, carboxymethyl-CoA, could not be detected in the mutant protein indicating that the lack of catalytic competence is due primarily to the inability of the protein to bind the second substrate, acetyl-CoA.


Transposon-tagging identifies novel pathogenicity genes in Fusarium graminearum.
With the increase of sequenced fungal genomes, high-throughput methods for functional analyses of genes are needed. We assessed the potential of a new transposon mutagenesis tool deploying a Fusarium oxysporum miniature inverted-repeat transposable element mimp1, mobilized by the transposase of impala, a Tc1-like transposon, to obtain knock-out mutants in Fusarium graminearum. We localized 91 mimp1 insertions which showed good distribution over the entire genome. The main exception was a major hotspot on chromosome 2 where independent insertions occurred at exactly the same nucleotide position. Furthermore insertions in promoter regions were over-represented. Screening 331 mutants for sexual development, radial growth and pathogenicity on wheat resulted in 19 mutants (5.7%) with altered phenotypes. Complementation with the original gene restored the wild-type phenotype in two selected mutants demonstrating the high tagging efficiency. This is the first report of a MITE transposon tagging system as an efficient mutagenesis tool in F. graminearum.


[Hodgkin's disease: effect of spenectomy on the immune status (author's transl)].
In 22 untreated Hodgkin's patients the following parameters were studied before and after splenectomy: the total WBC, lymphocytes, B- and T-cells and mitogenic stimulation of peripheral lymphocytes using autologous and control serum. The results were correlated to patient groups with favourable and unfavourable prognosis according to pathological stage, histology, spleen involvement and constitutional symptoms. 1. All Patients. Significant increase of the absolute number of peripheral lymphocytes and B-cells. Significant increase of the spontanous DNA synthesis in the presence of AB-control serum, but no change after mitogenic stimulation of peripheral lymphocytes. No change in the number of peripheral T-cells. 2. Patients with Favourable Prognosis. Significant increase of the absolute number of peripheral lymphocytes and B-cells. Significant increase of the spontanous DNA synthesis in the presence of AB-control-serum. Significant decrease of the T-cell function (PHA- and Con-A-stimulation in the presence of autologous serum). Significant decrease of the PHA-, Con-A- and PWM stimulation rate using control serum in the lymphocytic predominance and nodular sclerosis group. No change in the number of peripheral T-cells. 3. Patients with Unfavourable Prognosis. No change in the absolute number of B-cells, of the spontanous DNA-synthesis using autologous serum and of the PHA- and Con-A-stimulation. Significant increase of the EBV-induced blastogenesis in the mixed cellularity and lymphocytic depletion group.


Remora-Inspired Reversible Adhesive for Underwater Applications.
The remoras are marine species that can effectively move by clinging onto other marine species via a suction disk on their dorsal side, which is composed of complex structures. The inner suction disk could be divided into three large parts, namely, lip, lamella, and spinule. The lamella is deformed actively to generate pressure difference between the inside and outside of the suction disk, and the lip maintains the sealing. The spinule, which is composed of hairs with diameter of 300 μm or less hair, enhances the frictional force. In this study, we easily fabricated polymer-based adhesive inspired by the suction disk of the remora and conducted an experiment to determine its performance. The adhesive exhibited admirable performance with a 26.68 N cm-2 (266.8 kPa) pull-off strength and a 19.42 N cm-2 (194.2 kPa) shear strength in water. The durability test result indicated that the adhesion and friction characteristics of the adhesive were well maintained even after multiple uses.


Characterization of de novo diffuse large B-cell lymphoma with a translocation of c-myc and immunoglobulin genes.
The characteristics of de novo diffuse large B-cell lymphoma (DLBCL) with translocation of c-myc and immunoglobulin (Ig) genes (c-myc/Ig DLBCL), were investigated in 13 cases of c-myc/Ig DLBCL. Immunohistochemistry revealed five cases were positive for CD10 and BCL6 expression (CD10(+)/BCL6(+)), five cases of CD10(-)/BCL6(+)/MUM1(-), one case of CD10(-)/BCL6(+)/MUM1(+) and two cases of CD10(-)/BCL6(-)/MUM1(+) expression, indicating 10 cases of germinal center B-cell DLBCL and three cases of non-germinal center B-cell DLBCL. Ongoing mutation of the Ig heavy chain gene variable region (IgH-V) was found in two cases with CD10 and BCL6 expression and one case showing CD10(-)/BCL6(+)/MUM1(-) expression. These three cases of ongoing mutation of the IgH-V gene did not express BCL2, unlike those without ongoing mutation. These results suggest a heterogeneous immunophenotype and genotype for c-myc/Ig DLBCL, with CD10(-)/BCL6(+)/MUM1(-) cases the most frequent.


7-Chloro-4-(Phenylselanyl) Quinoline with Memory Enhancer Action in Aging Rats: Modulation of Neuroplasticity, Acetylcholinesterase Activity, and Cholesterol Levels.
This study investigated the effect of 7-chloro-4-(phenylselanyl) quinoline (4-PSQ) to restore the cognitive impairment caused by aging in male Wistar rats. Moreover, modulation of neuroplasticity markers, acetylcholinesterase (AChE) activity, and cholesterol levels was performed. Aged rats were intragastrically treated with 4-PSQ (5 mg/kg) for 7 days. Animals were tested in behavioral tasks, and then plasma (to determine cholesterol levels), hippocampus, and cerebral cortex (to determine neural cell adhesion molecule (NCAM) and polysialyltransferase (PST) levels, and AChE activity) were removed. Our findings demonstrated that treatment of aged rats with 4-PSQ restored short-term and long-term memories in the object recognition tests. 4-PSQ treatment did not restore exploratory activity (rearings) but partially restored locomotor activity (crossings) reduced by aging in the open-field test. Moreover, the compound restored the reduction in the NCAM and PST levels, and AChE activity in cerebral structures, as well as the increase in the plasma cholesterol levels, caused by aging in rats. In conclusion, 4-PSQ restored cognitive impairment caused by aging in rats by modulating synaptic plasticity, cholinergic system, and cholesterol levels.


[Ways to an efficient and practicable hospital hygiene from the point of view of a hospital hygienist (author's transl)].
Hospitalepidemiology means surveillance, prevention andocntrol of nosocomial infections. Trying to succeed he has to search for possiblities which are both practical as well as efficient: 1. The infection control nurse (one for 300 beds), 2. a bacteriological labor is for the epidemiologist, which is able to perform routine control on certain areas in the hospital (kitchen, sterilisation etc.), 3. encironmental examinations if necessary to find sources and for teaching purposes, 4. training of hospital personal in prevention, recognizing nosocomial infections, performing methods of desinfections etc., 5. trying to cooperate with the clinician in chemotherapy (selection of antibiotics, prophylaxis etc.), 6. to develop a programm to collect datas about nosocomial infections by a computer and to analyse those datas afterwards, 7. collaborativ work in a infection control commitee.


Assessment of pain and analgesic use in African American cancer patients: factors related to adherence to analgesics.
This study describes pain experience, analgesic use and barriers to pain control in African American cancer patients (N = 116). The overall adherence rate of analgesics was 46%. Constipation and nausea were the most commonly cited side effects of analgesics. Eighty-seven percent of patients reported concern about addiction to analgesics. Patients who believed their doctor needed to focus on curing illness rather than on controlling pain tended to comply with analgesic prescriptions (r = 0.20, p < 0.05). Patients with concerns that analgesics may cause confusion were less likely to take any type of analgesics (r = -0.16, p < 0.05). The study confirms that a patient's perceived barriers influence their decision to take analgesics, and also suggests that African American cancer patients may benefit from education that prevents misconceptions about analgesic use.


Bakuchiol inhibits lung cancer by modulating tumor microenvironment and the expression of PD-L1.
Immune checkpoint therapy is an emerging frontier in cancer therapy. With the aim to develop an efficient herb derived compound to facilitate immune checkpoint therapy, here we investigate if a herb-derived compound, Bakuchiol (BAK), can be used to treat lung cancer and elucidate if BAK could serve as a PD-L1 regulator. To this end, a murine lung cancer model was established by subcutaneously inoculating murine Lewis lung carcinoma (LLC) cells. BAK of 5 to 40 mg/kg was used for treatment in vivo for 15 days. On Day 15, the population of CD4+ and CD8+ T cells, Treg cells. BAK could effectively inhibit tumor growth by starting treatment either on Day 0 or 6 after tumor inoculation at doses of 5-40 mg/kg. BAK treatment increased the population of cytotoxic immune cells (i.e., CD8+ T cells, and M1 macrophages), meanwhile decreasing pro-tumor immune cells (i.e., CD3+ T cells, Treg cells, and M2 macrophages). Anti-inflammatory cytokines, including IL1β, IL2, IFNγ, TNF-α, IL4 and IL10 were upregulated by BAK. PD-L1 expression in the tumor was also lowered by BAK. AKT and STAT3 signaling were inhibited by BAK. BAK is an efficient agent in reducing LLC tumor growth. These data support the potential of BAK as a new drug for treating lung cancer by serving as a PD-L1 inhibitor that suppresses the activation of AKT and STAT3.


Neuroprotective effects of adrenomedullin in experimental traumatic brain injury model in rats.
Traumatic brain injuries cause damages in the brain in several ways, which include cell death because of edema, disruption of the blood-brain barrier, shear stress, and ischemia. In this study, we investigated the effects of adrenomedullin (AM) on oxidative stress and inflammation after head traumas in a rat model. Eighteen male adult Wistar albino rats were randomized into three groups (n=6). No traumas were applied to the con-trol (C) group. Traumas were applied in line with Marmarau trauma model in the trauma group. The rats in the AM treatment group were treated with post-traumatic 12 μg/kg i.p. AM in addition to the trauma group. The rats were followed for 7 days in all groups and were then sacrificed. Brain tissues and blood samples were taken. In the trauma group, both tissue and serum MDA, TNF-α, and IL-6 levels were significantly increased compared to the control group (p<0.05). In the AM-treated group, serum TNF-α levels were significantly decreased compared to the trauma group (p<0.05). In the trauma group, both tissue and serum GSH levels were significantly decreased compared to the control group (p<0.05). In the trauma group, serum Vitamin D3 levels were significantly decreased compared to the control group (p<0.05). In the AM-treated group, both tissue and serum GSH levels were significantly increased compared to the trauma group (p<0.05). These results indicate that AM has neuroprotective effects on traumatic brain injury in a rat model.


Chitosan-solid lipid nanoparticles as carriers for topical delivery of tretinoin.
Tretinoin (TRE) or all-trans retinoic acid is employed in the topical treatment of various skin diseases including acne and psoriasis. However, its use is strongly limited by side effects and high chemical instability. TRE encapsulation in nanostructured systems reduces these problems. Chitosan is a biopolymer that exhibits a number of interesting properties such as bioadhesion and antibacterial activity. The aim of this work was to prepare and characterize solid lipid nanoparticles (SLN) containing TRE, with and without addition of chitosan, to assess their in vitro cytotoxicity in keratinocytes and to evaluate their antibacterial activity against bacteria related to acne. SLN without (SLN-TRE) and with (SLN-chitosan-TRE) chitosan were prepared by hot high pressure homogenization. The hydrodynamic mean diameter and zeta potential were 162.7±1.4 nm and -31.9±2.0 mV for SLN-TRE, and 284.8±15.0 nm and 55.9±3.1 mV for SLN-chitosan-TRE. The SLN-chitosan-TRE exhibited high encapsulation efficiency, high physical stability in the tested period (one year), were not cytotoxic to keratinocytes and showed high antibacterial activity against P. acnes and S. aureus. Therefore chitosan-SLN can be good candidates to encapsulate TRE and to increase its therapeutic efficacy in the topical treatment of acne.


Effects of exogenous organic/inorganic nitrogen addition on carbon pool distribution and transformation in grassland soil.
Increases in nitrogen (N) deposition may significantly affect the organic carbon (OC) cycle in soil. The inconsistent findings of the influence of added N on soil OC pools highlight the need of quantifying responses of the OC pool distribution to N addition. Moreover, the influence of N addition with a mixture of organic and inorganic N on OC pool distribution and stabilization in grassland soil remains unclear. We carried out a five-year field experiment with adding N to examine the effects of different types of N addition on soil OC pool distribution and transformation in a meadow steppe in Inner Mongolia. We applied N in the ratios of inorganic N (IN) and organic N (ON) at 10:0 (N1), 7:3 (N2), 5:5 (N3), 3:7 (N4), 0:10 (N5), and 0:0 (CK), respectively. We measured OC content in bulk soil, particulate organic matter (POM), and mineral-associated organic matter (MAOM) fractions. Additionally, a short-term soil incubation was conducted to assess potential OC mineralization. Our study showed no significant effect on soil organic carbon content of different ratios of IN/ON addition. N addition reduced microbial biomass C/N ratio, the fraction of mineral-associated organic matter, cumulative CO2 emission, and microbial metabolic quotient. Compared with ON addition alone, IN addition alone showed a stronger effect on the C in different soil fractions and soil OC mineralization. The particulate organic matter (POM) fraction was more sensitive to N addition than the mineral-associated organic matter (MAOM) fraction. Our results suggest that the contribution of N in organic and inorganic forms affecting OC pool distribution with different turnover rates should be considered when assessing the effects of N addition types on soil OC processes in grassland.


Longitudinal correlates of fruit and vegetable consumption with depressive symptoms among middle-aged and older adults in South Africa.
Fruit and vegetable intake may influence mental well-being. The aim of this study was to assess longitudinal associations between fruit and vegetable intake and depressive symptoms among rural South Africans. This longitudinal community study enrolled 3,891 adults (≥ 40 years) from the "Health and Ageing in Africa: A Longitudinal Study of an INDEPTH Community in South Africa (HAALSI)". Fruit and vegetable intake was assessed by self-report at wave 1, and depressive symptoms were assessed using the Center for Epidemiological Studies Depression Scale at wave 1 and 2. Outcomes were incident and persistent depressive symptoms at wave 2. Logistic regression was used to assess the associations between fruit and vegetable intake at wave 1 and incident, and persistent depressive symptoms. Results indicate that in the fully adjusted model for individuals with no depressive symptoms at baseline, we found no significant association between frequency of fruit or vegetables intake and incident depressive symptoms. We also found no significant association between frequency of fruit or vegetable intake and persistent depressive symptoms. Fruit and vegetable intake was not significantly associated with incident and persistent depressive symptoms.


Diagnosis of neurosyphilis in HIV-negative patients with syphilis: development, validation, and clinical utility of a suite of machine learning models.
The ability to accurately identify the absolute risk of neurosyphilis diagnosis for patients with syphilis would allow preventative and therapeutic interventions to be delivered to patients at high-risk, sparing patients at low-risk from unnecessary care. We aimed to develop, validate, and evaluate the clinical utility of simplified clinical diagnostic models for neurosyphilis diagnosis in HIV-negative patients with syphilis. We searched PubMed, China National Knowledge Infrastructure and UpToDate for publications about neurosyphilis diagnostic guidelines in English or Chinese from database inception until March 15, 2023. We developed and validated machine learning models with a uniform set of predictors based on six authoritative diagnostic guidelines across four continents to predict neurosyphilis using routinely collected data from real-world clinical practice in China and the United States (through the Dermatology Hospital of Southern Medical University in Guangzhou [659 recruited between August 2012 and March 2022, treated as Development cohort], the Beijing Youan Hospital of Capital Medical University in Beijng [480 recruited between December 2013 and April 2021, treated as External cohort 1], the Zhongshan Hospital of Xiamen University in Xiamen [493 recruited between November 2005 and November 2021, treated as External cohort 2] from China, and University of Washington School of Medicine in Seattle [16 recruited between September 2002 and April 2014, treated as External cohort 3] from United States). We included all these patients with syphilis into our analysis, and no patients were further excluded. We trained eXtreme gradient boosting (XGBoost) models to predict the diagnostic outcome of neurosyphilis according to each diagnostic guideline in two scenarios, respectively. Model performance was measured through both internal and external validation in terms of discrimination and calibration, and clinical utility was evaluated using decision curve analysis. The final simplified clinical diagnostic models included neurological symptoms, cerebrospinal fluid (CSF) protein, CSF white blood cell, and CSF venereal disease research laboratory test/rapid plasma reagin. The models showed good calibration with rescaled Brier score of 0.99 (95% CI 0.98-1.00) and excellent discrimination (the minimum value of area under the receiver operating characteristic curve, 0.84; 95% CI 0.81-0.88) when externally validated. Decision curve analysis demonstrated that the models were useful across a range of neurosyphilis probability thresholds between 0.33 and 0.66 compared to the alternatives of managing all patients with syphilis as if they do or do not have neurosyphilis. The simplified clinical diagnostic models comprised of readily available data show good performance, are generalisable across clinical settings, and have clinical utility over a broad range of probability thresholds. The models with a uniform set of predictors can simplify the sophisticated clinical diagnosis of neurosyphilis, and guide decisions on delivery of neurosyphilis health-care, ultimately, support accurate diagnosis and necessary treatment. The Natural Science Foundation of China General Program, Health Appropriate Technology Promotion Project of Guangdong Medical Research Foundation, Department of Science and technology of Guangdong Province Xinjiang Rural Science and Technology(Special Commissioner)Project, Southern Medical University Clinical Research Nursery Garden Project, Beijing Municipal Administration of Hospitals Incubating Program.


Improvement of thermostability and activity of pectate lyase in the presence of hydroxyapatite nanoparticles.
The activity and half-life of pectate lyase (PL) from Bacillus megaterium were nine- and 60-fold, respectively, higher at 90 °C in the presence of hydroxyapatite nanoparticles (NP-PLs) than in the presence of 1mM CaCl(2). Thermodynamic analysis of the nanoparticle-induced stability revealed an enhanced entropy-enthalpy compensation by the NP-PLs since a reciprocal linearity of the enthalpy-entropy change to 90 °C was observed. Without nanoparticles, the linearity range was 70 °C. Such compensation reflected the maintenance of the native structure of proteins. The remarkable enhancement of activity and stability of the NP-PL system at high temperatures may be utilized commercially e.g. in the food industry or the processing of natural fibers that may require a thermotolerant enzyme.


Primary standard for liquid flow rates between 30 and 1500 nl/min based on volume expansion.
An increasing number of microfluidic systems operate at flow rates below 1 μl/min. Applications include (implanted) micropumps for drug delivery, liquid chromatography, and microreactors. For the applications where the absolute accuracy is important, a proper calibration is required. However, with standard calibration facilities, flow rate calibrations below ~1 μl/min are not feasible because of a too large calibration uncertainty. In the current research, a traceable flow rate using a certain temperature increase rate is proposed. When the fluid properties, starting mass, and temperature increase rate are known, this principle yields a direct link to SI units, which makes it a primary standard. In this article, it will be shown that this principle enables flow rate uncertainties in the order of 2-3% for flow rates from 30 to 1500 nl/min.


Vascular and nonvascular intracranial malformation associated with external capillary hemangiomas.
Seven patients with facial and scalp capillary hemangiomas with associated and nonvascular intracranial malformations are presented. All patients were female. The most important anomalies include: Dandy-Walker disease, cerebellar hypoplasia, arterial angiomas, and abnormal origin and intracranial distribution of some of the principal cerebral arteries.


Weakly Supervised AI for Efficient Analysis of 3D Pathology Samples.
Human tissue consists of complex structures that display a diversity of morphologies, forming a tissue microenvironment that is, by nature, three-dimensional (3D). However, the current standard-of-care involves slicing 3D tissue specimens into two-dimensional (2D) sections and selecting a few for microscopic evaluation1,2, with concomitant risks of sampling bias and misdiagnosis3-6. To this end, there have been intense efforts to capture 3D tissue morphology and transition to 3D pathology, with the development of multiple high-resolution 3D imaging modalities7-18. However, these tools have had little translation to clinical practice as manual evaluation of such large data by pathologists is impractical and there is a lack of computational platforms that can efficiently process the 3D images and provide patient-level clinical insights. Here we present Modality-Agnostic Multiple instance learning for volumetric Block Analysis (MAMBA), a deep-learning-based platform for processing 3D tissue images from diverse imaging modalities and predicting patient outcomes. Archived prostate cancer specimens were imaged with open-top light-sheet microscopy12-14 or microcomputed tomography15,16 and the resulting 3D datasets were used to train risk-stratification networks based on 5-year biochemical recurrence outcomes via MAMBA. With the 3D block-based approach, MAMBA achieves an area under the receiver operating characteristic curve (AUC) of 0.86 and 0.74, superior to 2D traditional single-slice-based prognostication (AUC of 0.79 and 0.57), suggesting superior prognostication with 3D morphological features. Further analyses reveal that the incorporation of greater tissue volume improves prognostic performance and mitigates risk prediction variability from sampling bias, suggesting that there is value in capturing larger extents of spatially heterogeneous 3D morphology. With the rapid growth and adoption of 3D spatial biology and pathology techniques by researchers and clinicians, MAMBA provides a general and efficient framework for 3D weakly supervised learning for clinical decision support and can help to reveal novel 3D morphological biomarkers for prognosis and therapeutic response.


Determination of corrections to true summations of photons for measurements in Marinelli beakers.
This paper describes a method of determination of correction factors for the true summation effect of photons for measurement of activity of radionuclides emitting gamma radiation. The method was tested with Marinelli beakers samples measured with coaxial germanium detectors. The advantage of this method is its simplicity since it is not necessary to determine the curves of total efficiency for each detector. The correction factor for the particular peak is determined by means of a detector index which is calculated from one of the measurements with a calibration standard.


SIRT3 Is a Critical Regulator of Mitochondrial Function of Fibroblasts in Pulmonary Hypertension.
Pulmonary hypertension (PH) is a heterogeneous and life-threatening cardiopulmonary disorder in which mitochondrial dysfunction is believed to drive pathogenesis, although the underlying mechanisms remain unclear. To determine if abnormal SIRT3 (sirtuin 3) activity is related to mitochondrial dysfunction in adventitial fibroblasts from patients with idiopathic pulmonary arterial hypertension (IPAH) and hypoxic PH calves (PH-Fibs) and whether SIRT3 could be a potential therapeutic target to improve mitochondrial function, SIRT3 concentrations in control fibroblasts, PH-Fibs, and lung tissues were determined using quantitative real-time PCR and western blot. SIRT3 deacetylase activity in cells and lung tissues was determined using western blot, immunohistochemistry staining, and immunoprecipitation. Glycolysis and mitochondrial function in fibroblasts were measured using respiratory analysis and fluorescence-lifetime imaging microscopy. The effects of restoring SIRT3 activity (by overexpression of SIRT3 with plasmid, activation SIRT3 with honokiol, and supplementation with the SIRT3 cofactor nicotinamide adenine dinucleotide [NAD+]) on mitochondrial protein acetylation, mitochondrial function, cell proliferation, and gene expression in PH-Fibs were also investigated. We found that SIRT3 concentrations were decreased in PH-Fibs and PH lung tissues, and its cofactor, NAD+, was also decreased in PH-Fibs. Increased acetylation in overall mitochondrial proteins and SIRT3-specific targets (MPC1 [mitochondrial pyruvate carrier 1] and MnSOD2 [mitochondrial superoxide dismutase]), as well as decreased MnSOD2 activity, was identified in PH-Fibs and PH lung tissues. Normalization of SIRT3 activity, by increasing its expression with plasmid or with honokiol and supplementation with its cofactor NAD+, reduced mitochondrial protein acetylation, improved mitochondrial function, inhibited proliferation, and induced apoptosis in PH-Fibs. Thus, our study demonstrated that restoration of SIRT3 activity in PH-Fibs can reduce mitochondrial protein acetylation and restore mitochondrial function and PH-Fib phenotype in PH.


Comparison of Fusobacterium sp. versus Bacteroides sp. in the monoxenic cultivation of E. histolytica.
By means of the performed studies it was established that 6 Fusobacteria strains enhanced the growth of E. histolytica while 4 Bacteroides strains did not possess this property.


Surface functionalization of doxorubicin-loaded liposomes with octa-arginine for enhanced anticancer activity.
Doxorubicin-loaded PEGylated liposomes (commercially available as DOXIL or Lipodox) were surface functionalized with a cell-penetrating peptide, octa-arginine (R8). For this purpose, R8-peptide was conjugated to the polyethylene glycol-dioleoyl phosphatidylethanolamine (PEG-DOPE) amphiphilic co-polymer. The resultant R8-PEG-PE conjugate was introduced into the lipid bilayer of liposomes at 2 mol% of total lipid amount via spontaneous micelle-transfer technique. The liposomal modification did not alter the particle size distribution, as measured by Particle Size Analyzer and transmission electron microscopy (TEM). However, surface-associated cationic peptide increased zeta potential of the modified liposomes. R8-functionalized liposomes (R8-Dox-L) markedly increased the intracellular and intratumoral delivery of doxorubicin as measured by flow cytometry and visualizing by confocal laser scanning microscopy (CLSM) compared to unmodified Doxorubicin-loaded PEGylated liposomes (Dox-L). R8-Dox-L delivered loaded Doxorubicin to the nucleus, being released from the endosomes at higher efficiency compared to unmodified liposomes, which had marked entrapment in the endosomes at tested time point of 1h. The significantly higher accumulation of loaded drug to its site of action for R8-Dox-L resulted in improved cytotoxic activity in vitro (cell viability of 58.5 ± 7% for R8-Dox-L compared to 90.6 ± 2% for Dox-L at Dox dose of 50 μg/mL for 4h followed by 24h incubation) and enhanced suppression of tumor growth (348 ± 53 mm(3) for R8-Dox-L, compared to 504 ± 54 mm(3) for Dox-L treatment) in vivo compared to Dox-L. R8-modification has the potential for broadening the therapeutic window of pegylated liposomal doxorubicin treatment, which could lead to lower non-specific toxicity.


Better prognostic ability of NEWS2, SOFA and SAPS-II in septic patients.
To compare the ability of qSOFA, NEWS2, SOFA, LODS, SIRS, APACHE-II and SAPS-II scores. Analysis of in-hospital mortality of 203 patients admitted to the ICU because of sepsis. The scores were compared according to their application. Discrimination was evaluated with AUC-ROC curve and performance with the Akaike's (AIC) and Bayesian information criterion (BIC). In-hospital mortality was 31.53%. NEWS2 showed better mortality discrimination ability and better performance considering the AIC/BIC criterion for mortality tan qSOFA (AUC-ROC=.615 and .536; P=.039). SOFA presented higher performance and AUC-ROC tan LODS (.776 vs .693; P=.01) and both showed higher discrimination ability than SIRS (AUC-ROC=.521; P<.003). Finally, SAPS-II was able to predict mortality with better performance than APACHE-II and presented higher discrimination capacity but without statistical significance compared (AUROC=.738 for SAPS-II and AUROC=.673 for APACHE-II; P=.08). NEWS2 is a better predictor of mortality than qSOFA and its implementation for the early recognition of the septic patient or the patient with higher risk in the emergency and hospitalization wards should be addressed. In addition, given that SOFA and SAPS-II showed better performance and are simpler than LODS and APACHE-II, respectively, both should be considered the scores of choice in this setting.


Human chorionic gonadotropin attenuates the vascular response to angiotensin II.
Our purpose was to investigate interactions between human chorionic gonadotropin (hCG) and angiotensin II (ANG II) in resistance arteries. Isolated pre-arteriolar vessels of female Sprague-Dawley rats were prepared from the mesenteric arcade and the mesometrium. Dose-response curves were recorded by means of an video-electronic arteriograph system. (1) hCG dilated mesenteric (MA; EC(50): 21+/-6.9 mU/ml) and uterine (UA; EC(50): 256+/-44.3 mU/ml)) resistance arteries pre-constricted with ANG II. In the presence of glybenclamide (1 micromol/l), the response of MA was reduced by >50%. (2) After application of hCG for 1h, vasoconstriction by ANG II was significantly attenuated in MA. This effect was dose-dependent and was not observed in UA. Efficacy of ANG II could be restored, when hCG was followed by glybenclamide. hCG may contribute to the reduction in arterial tone seen early in human pregnancy. Its vascular effects are in part mediated by the activation of adenosine 5'-triphosphate-sensitive potassium channels, suggesting a protein kinase A dependent signaling pathway.


Glycosyltransferases with endogenous acceptor activity in plasma membranes isolated from rat liver.
Plasma membrane fractions from rat liver exhibited glycosyltransferase activity with endogenous membrane-associated acceptors and either UDP-galactose, UDPglucose, UDP-N-acetylglucosamine, or GDPmannose donors. Of these, incorporation into non-lipid acceptors was most active with UDP-galactose and only with UDPgalactose and UDPmannose was there incorporation into endogenous lipid acceptors. CMP-N-acetylneuraminic acid was inactive as a donor with the isolated plasma membranes. In order to demonstrate transferase activity, low concentrations of substrate sugar nucleotides and short incubation times were used as well as sulfhydryl protectants and a phosphatase inhibitor (NaF) in the reaction mixtures. The findings support the concept of surface localization of at least a galactosyl transferase in cells of rat liver.


Identification of odorant-binding protein genes from antennal expressed sequence tags of the onion fly, Delia antiqua.
Insect odorant-binding proteins (OBPs) are thought to play a crucial role in the chemosensation of hydrophobic molecules such as pheromones and host chemicals. The onion fly, Delia antiqua, is a specialist feeder of Allium plants, and utilizes a host odorant n-dipropyl disulfide as a cue for its oviposition. Because n-dipropyl disulfide is a highly hydrophobic compound, some OBPs might be indispensable for perception of it. However, no OBP gene has been identified in D. antiqua. Here, to obtain the DNA sequences of D. antiqua OBPs, we performed an analysis of antennal expressed sequence tags (ESTs). Among 288 EST clones, eight D. antiqua OBP genes were identified for the first time. Phylogenetic analysis revealed that each D. antiqua OBP gene is more closely related to its Drosophila orthologs than to the other D. antiqua OBP genes, suggesting that these OBP genes had emerged before the divergence of Delia and Drosophila species. All of the eight D. antiqua OBPs are expressed not only in the antennae but also in the legs, suggesting additional roles in the taste perception of non-volatile compounds. These findings serve as an important basis for understanding the molecular mechanisms underlying the host adaptations of D. antiqua.


Glucose 6-P dehydrogenase delays the onset of frailty by protecting against muscle damage.
Frailty is a major age-associated syndrome leading to disability. Oxidative damage plays a significant role in the promotion of frailty. The cellular antioxidant system relies on reduced nicotinamide adenine dinucleotide phosphate (NADPH) that is highly dependent on glucose 6-P dehydrogenase (G6PD). The G6PD-overexpressing mouse (G6PD-Tg) is protected against metabolic stresses. Our aim was to examine whether this protection delays frailty. Old wild-type (WT) and G6PD-Tg mice were evaluated longitudinally in terms of frailty. Indirect calorimetry, transcriptomic profile, and different skeletal muscle quality markers and muscle regenerative capacity were also investigated. The percentage of frail mice was significantly lower in the G6PD-Tg than in the WT genotype, especially in 26-month-old mice where 50% of the WT were frail vs. only 13% of the Tg ones (P < 0.001). Skeletal muscle transcriptomic analysis showed an up-regulation of respiratory chain and oxidative phosphorylation (P = 0.009) as well as glutathione metabolism (P = 0.035) pathways in the G6PD-Tg mice. Accordingly, the Tg animals exhibited an increase in reduced glutathione (34.5%, P < 0.01) and a decrease on its oxidized form (-69%, P < 0.05) and in lipid peroxidation (4-HNE: -20.5%, P < 0.05). The G6PD-Tg mice also showed reduced apoptosis (BAX/Bcl2: -25.5%, P < 0.05; and Bcl-xL: -20.5%, P < 0.05), lower levels of the intramuscular adipocyte marker FABP4 (-54.7%, P < 0.05), and increased markers of mitochondrial content (COX IV: 89.7%, P < 0.05; Grp75: 37.8%, P < 0.05) and mitochondrial OXPHOS complexes (CII: 81.25%, P < 0.01; CIII: 52.5%, P < 0.01; and CV: 37.2%, P < 0.05). Energy expenditure (-4.29%, P < 0.001) and the respiratory exchange ratio were lower (-13.4%, P < 0.0001) while the locomotor activity was higher (43.4%, P < 0.0001) in the 20-month-old Tg, indicating a major energetic advantage in these mice. Short-term exercise training in young C57BL76J mice induced a robust activation of G6PD in skeletal muscle (203.4%, P < 0.05), similar to that achieved in the G6PD-Tg mice (142.3%, P < 0.01). Glucose 6-P dehydrogenase deficiency can be an underestimated risk factor for several human pathologies and even frailty. By overexpressing G6PD, we provide the first molecular model of robustness. Because G6PD is regulated by pharmacological and physiological interventions like exercise, our results provide molecular bases for interventions that by increasing G6PD will delay the onset of frailty.


Ocular toxoplasmosis in pregnancy.
To describe the course of ocular toxoplasmosis during pregnancy. This study was a retrospective, non-comparative case series of four pregnant women who were treated for ocular toxoplasmosis during pregnancy. All of the participants had severe and treatment-resistant toxoplasmic retinochoroiditis during pregnancy, leaving three of them with decreased visual acuity in spite of aggressive therapy. Delivery of the infant appeared to help the recovery in two patients. Pregnant state may provoke the recurrence of ocular toxoplasmosis.


Antiviral isoflavonoid sulfate and steroidal glycosides from the fruits of Solanum torvum.
The C-4 sulfated isoflavonoid, torvanol A (1), and the steroidal glycoside, torvoside H (3), together with the known glycoside, torvoside A (2), were isolated from a MeOH extract of Solanum torvum fruits. Upon enzymatic hydrolysis with beta-glucosidase, torvoside A (2) and torvoside H (3) yielded the corresponding acetal derivatives 4 and 5, respectively. Torvanol A (1), torvoside H (3) and compound 5 exhibited antiviral activity (herpes simplex virus type 1) with IC(50) values of 9.6, 23.2 and 17.4 microg/ml, respectively. Compounds 1-5 showed no cytotoxicity (at 50 microg/ml) against BC, KB and Vero cell lines.


Nodal metastases in small rectal neuroendocrine tumours.
Rectal neuroendocrine tumours (NETs) are the most common type of gastrointestinal NET. European Neuroendocrine Tumour Society guidelines suggest that rectal NETs measuring ≤10 mm are indolent with low risk of spread. In practice, many patients with lesions ≤1 cm do not undergo complete tumour staging. However, the size of the lesion may not be the only risk factor for nodal involvement/metastases. The aim of this study was to determine if MRI ± nuclear medicine imaging alters tumour stage in patients with rectal NETs ≤10 mm. Patients referred to a tertiary NET centre between 2005 and 2020 who met the inclusion criteria of a rectal NET ≤10 mm, full cross-sectional imaging, primarily an MRI scan and, if abnormal findings were identified, a subsequent 68 Ga-DOTATATE positron emission tomography scan were included. All patients were followed up at our institution. In all, 32 patients with rectal NETs 10 mm or less were included in the study: 16 women; median age 58 years (range 33-71); 47% (n = 15) were referred from bowel cancer screening procedures. The median size of the lesions was 5 mm (range 2-10 mm). 81% (n = 26) were World Health Organization Grade 1 tumours with Ki67 <3%. Radiological staging confirmed nodal involvement in 25% (8/32); two cases had distant metastatic disease. Lymphovascular invasion was present in 3% (1/32) of patients but none demonstrated peri-neural invasion. This study demonstrates that small rectal NETs can develop nodal metastases; therefore it is important to stage these tumours accurately with MRI at baseline and, if there are concerns regarding potential lymph node metastases, to consider 68 Ga-DOTATATE positron emission tomography imaging.


Isolation and bioactivity of new tanshinones.
Two new diterpenoids, designated neocryptotanshinone and isotanshinone IIB, have been isolated from "Tan-Shen," the root of Salvia miltiorrhiza, together with a known compound, danshexinkun A. Their structures are established by spectral and physical data. Isotanshinone IIB exhibits significant inhibitory activity in vitro on ADP- and collagen-induced aggregation.


Regional differences in outcome from subarachnoid haemorrhage.
Surgeons are increasingly placed under pressure to accept publication of their results and to abide by recommendations to change practice which others derive. Considerable concern exists about misinterpretation of such data. The issue is well illustrated by this study. Data on outcome following treatment for subarachnoid haemorrhage were prospectively collected from 1993-1998 in two centres in the British Isles: Newcastle and Nottingham. Initial examination of this data suggest a substantial difference in the performance favouring Nottingham over Newcastle. The odds of a poor outcome was 1:1.86 in Newcastle compared with 1:4.26 in Nottingham giving an odds ratio of 2.3 in favour of Nottingham and this difference was highly significant with p<0.00001. On a more detailed examination taking account of confounding variables, this difference disappeared entirely. Newcastle was able to operate a less selective admissions policy than Nottingham because of the deficiency of beds at the latter unit. A summary of these results has been published elsewhere. These results illustrate the dangers of applying statistical tools developed for simpler situations such as industrial process control to complex medical problems. We conclude that comprehensive and accurate data on all factors likely to influence the outcome for a particular treatment should be collected as an absolute prerequisite to any judgments being made on apparent statistical differences between the performances of differing units.


Aluminum-based temporary removable partial dentures.
The fabrication of an aluminum-based temporary removable partial denture has been presented. The advantages of this type of prosthesis include compatibility with oral tissues, accuracy of fit, decreased bulk, and improved thermal conductivity.


Clinical and economic challenges facing pharmacogenomics.
In this paper, we examine the clinical and economic challenges that face developers of and payers for personalized drugs and companion diagnostics. We review and summarize clinical, regulatory and reimbursement issues with respect to eight, high profile personalized medicines and their companion diagnostics. Subsequently, we determine Medicare parts B and D reimbursement of the eight drugs from publicly available databases. Finally, we utilize surveys-each tailored to three key stakeholders; payers, drug and diagnostic developers, and pharmacogenomic expert analysts-to assess reimbursement of diagnostics, analyze the role that different kinds of evidence have in informing prescribing and reimbursement decisions, as well as the specific clinical, regulatory and economic challenges that confront pharmacogenomics as it moves forward. We found that Medicare beneficiary access to physician-administered (Medicare part B) drugs is relatively unfettered, with a fixed patient co-insurance percentage of 20%. More reimbursement restrictions are placed on self-administered (Medicare part D) drugs, which translates into higher and more variable cost sharing, more use of prior authorization and quantity limits. There is a lack of comprehensive reimbursement of companion diagnostics, even in cases in which the diagnostic is on the label and recommended or required by the Food and Drug Administration. Lack of evidence linking diagnostic tests to health outcomes has caused payers to be skeptical about the clinical usefulness of tests. Expert analysts foresee moderate growth in post-hoc development of companion diagnostics to personalize already approved drugs, and limited growth in the concurrent co-development of companion diagnostics and personalized medicines. Lack of clinically useful diagnostics as well as an evidence gap in terms of knowledge of drug and diagnostic clinical effectiveness appear to be hindering growth in personalized medicine. An increase in comparative effectiveness research may help to close the evidence gap.


A study of non-isotopic in situ hybridization histochemistry on postmortem changes in vasopressin mRNA in rat brain.
Rat models which stimulated various postmortem conditions were used to determine postmortem changes in the vasopressin mRNA content of the brain. In situ hybridization histochemistry experiments using biotinylated oligonucleotide probes revealed that vasopressin mRNA could be detected in the rat hypothalamic nuclei, even though the rats had been killed and left for 8 h postmortem at room temperature and the brains were then fixed by immersion in 4% paraformaldehyde instead of by transcardial perfusion. However, these signals were greatly reduced in the nuclei if the brains were removed after a 24-h delay. These results suggest that gene expression study of neuropeptides at the cellular level can be performed on postmortem human brains after a short postmortem delay.


Elevated noise power in gamma band related to negative symptoms and memory deficit in schizophrenia.
There is an increasing consideration for a disorganized cerebral activity in schizophrenia, perhaps relating to a synaptic inhibitory deficit in the illness. Noise power (scalp-recorded electroencephalographic activity unlocked to stimuli) may offer a non-invasive window to assess this possibility. 29 minimally-treated patients with schizophrenia (of which 17 were first episodes) and 27 healthy controls underwent clinical and cognitive assessments and an electroencephalographic recording during a P300 paradigm to calculate signal-to-noise ratio and noise power magnitudes in the theta and gamma bands. In comparison to controls, a significantly higher gamma noise power was common to minimally-treated and first episode patients over P3, P4, T5 and Fz electrode sites. Those high values were directly correlated to negative symptom severity and inversely correlated to verbal memory scores in the patients. There were no differences in signal-to-noise ratio magnitudes among the groups. Gamma noise power at Fz discriminated significantly between patients and controls. No significant differences were found in theta noise power or in gamma noise power over the other electrode sites between the groups of patients and controls. We have not assessed phase-locked and non-phase locked power changes, a complementary approach that may yield useful information. Gamma noise power may represent a useful and non-invasive tool for studying brain dysfunction in psychotic illness. These results suggest an inefficient activation pattern in schizophrenia.


Comparison of synaptic transmission and plasticity between sensory and cortical synapses on relay neurons in the ventrobasal nucleus of the rat thalamus.
Relay neurons in the ventrobasal nucleus of the thalamus transmit somatosensory information to the cerebral cortex and receive sensory and cortical (feedback) synaptic inputs via, respectively, medial lemniscal (ML) and corticothalamic (CT) fibres. Here, we report that calcium-permeable AMPA receptors are expressed at CT synapses, but not ML synapses, and that the NMDA receptor (NMDAR)-mediated/non-NMDAR-mediated synaptic current ratio is significantly larger at CT synapses than at ML synapses. Moreover, NMDAR-dependent LTP and L-type voltage-gated calcium channel-dependent LTD are readily induced at CT synapses, but not ML synapses. In particular, LTD of CT synaptic transmission is induced by spiking of postsynaptic relay neurons in continuous mode, but not burst mode, in current-clamp recordings. These results show that the strength of the cortical input to thalamic relay neurons is selectively subjected to use-dependent modification, which could be a mechanism for regulation of thalamocortical-corticothalamic interactions and the underlying sensory processing.


Metabolomics characterizes early metabolic changes and markers of tolerant Eucalyptus ssp. clones against drought stress.
L'Hér. (Myrtaceae) is one of the economically most important and widely cultivated trees for wood crop purposes worldwide. Climatic changes together with the constant need to expand plantations to areas that do not always provide optimal conditions for plant growth highlight the need to assess the impact of abiotic stresses on eucalypt trees. We aimed to unveil the drought effect on the leaf metabolome of commercial clones with differential phenotypic response to this stress. For this, seedlings of 13 clones were grown at well-watered (WW) and water-deficit (WD) conditions and their leaf extracts were subjected to comparative analysis using ultra-high performance liquid chromatography coupled to mass spectrometry (UPLC-MS) and nuclear magnetic resonance spectroscopy (NMR). UPLC-MS and NMR analyses led to the annotation of over 100 molecular features of classes such as cyclitols, phenolics, flavonoids, formylated phloroglucinol compounds (FPCs) and fatty acids. Multivariate data analysis was employed for specimens' classifications and markers identification from both platforms. The results obtained in this work allowed us to classify clones differing in drought tolerance. Classification models were validated using an extra subset of samples. Tolerant plants exposed to water deficit accumulated arginine, gallic acid derivatives, caffeic acid and tannins at higher levels. In contrast, stressed drought-sensitive clones were characterised by a significant reduction in glucose, inositol and shikimic acid levels. These changes in contrasting drought response eucalypt pave ways for differential outcomes of tolerant and susceptible phenotypes. Under optimal growth conditions, all clones were rich in FPCs. These results can be used for early screening of tolerant clones and to improve our understanding of the role of these biomarkers in Eucalyptus tolerance to drought stress.


Treatment of the N0 neck during salvage surgery after radiotherapy of head and neck squamous cell carcinoma.
The morbidity and mortality rates of salvage surgery in patients with local recurrence of head and neck squamous cell carcinoma (HNSCC) after radiotherapy are high. The aim of this study was to determine the rate of occult neck node metastasis and the surgical morbidity of patients after salvage surgery for local relapse after definitive radiotherapy. Thirty patients who underwent salvage surgery with a simultaneous neck node dissection for a local relapse after definitive radiotherapy for HNSCC between 1992 and 2000 were included in this study. The primary tumor sites were oral cavity in six patients, oropharynx in 17, supraglottic larynx in three, and hypopharynx in four. Initially, seven patients had T2 disease, eight had T3, and 15 had T4. Twelve patients (40%) experienced postoperative complications, including two deaths. There was no cervical lymph node metastasis (pN0) in 29 of the 30 patients. Fifteen patients (50%) had a recurrence after salvage surgery, including 11 new local recurrences and four patients with distant metastasis. The risk of neck node metastasis during salvage surgery for local recurrence in patients treated initially with radiation for N0 HNSCC is low. Neck dissection should be performed in only limited area, depending on the surgical procedure used for tumor resection.


Efficient nonadhesive ex vivo expansion of early endothelial progenitor cells derived from CD34+ human cord blood fraction for effective therapeutic vascularization.
Endothelial progenitor cells (EPCs) have been shown to have therapeutic potential in ischemic disease. However, the number of EPCs for cell therapy is limited. In this study, instead of the typical adherent culture method, we investigated a more efficient, clinically applicable nonadhesive expansion method for early EPCs using cord blood-derived cells to overcome rapid cellular senescence. After a suspension culture of isolated CD34(+) cells in serum-free medium containing each cytokine combination was maintained for 9 d, the number of expanded functional EPCs was assessed by an adherent culture assay. Compared to mononuclear cells, the CD34(+) fraction was superior in its expansion of functional EPCs that could differentiate into acLDL/UEA-1(+) cells without significant cellular senescence, whereas the CD34(-) fraction showed no EPC expansion. Among the cytokine combinations tested for the CD34(+) fraction, a combination (SFIb) consisting of stem cell factor (SCF), FMS-like tyrosine kinase 3 ligand, interleukin-3, and basic fibroblast growth factor resulted in a reproducible 64- to 1468-fold EPC expansion from various cord blood origins. Interestingly, the SFIb combination displayed markedly increased EPC expansion (2.43-fold), with a higher percentage of CD34(+) cells (2.17-fold), undifferentiated blasts (2.38-fold) and CXCR4(+) cells (1.68-fold) compared to another cytokine combination (SCF, thrombopoietin, and granulocyte colony-stimulating factor), although the two cytokine combinations had a similar level of total mononucleated cell expansion (∼ 10% difference). Accordingly, the cells expanded in the SFIb combination were more effective in recovery of blood flow and neovascularization in hind-limb ischemia in vivo. Taken together, these results suggest that the nonadhesive serum-free culture conditions of the CD34(+) fraction provide an effective EPC expansion method for cell therapy, and an expansion condition leading to high percentages of CD34(+) cells and blasts is likely important in EPC expansion.


Comparison of urinary bone resorption markers in women of 40-70 years; day-to-day and long-term variation in individual subjects.
Bone resorption can be judged using biochemical markers in urine and blood. Our aim was to study the patterns of markers in the postmenopausal period. The urinary excretion of bone resorption markers was tested using different assays. The study was undertaken to determine the day-to-day and the long-term variation, over 8 years, of these markers in individual women. Over a period of 2 weeks, the median of the day-to-day variation of the pyridinium crosslink markers varied between 12 and 23%, the median value of the long-term variation over 8 years between 10 and 21%, for the telopeptide markers median day-to-day variation was 18 and 20% and the long-term variation was 17 and 19%. The correlations between the different crosslink markers varied between 0.63 and 0.92, depending on the kind of the crosslink and on the method of determination. The two telopeptide markers showed an excellent correlation with r of 0.95. The excretion of all bone resorption markers varied with postmenopausal age, some differences were found between the crosslink and the telopeptide excretions with age, in women more than 20 years postmenopausal the telopeptides decrease whereas the crosslinks show an increase. This study shows that crosslinks and telopeptides give similar information on the rate of bone resorption: an increase during the first 5 years and a slight decrease in the next 5 years after menopause, discrepancies were found after 10 or more postmenopausal years.


Leukotriene B4 as a marker of cerebral dysfunction.
It has been postulated that leukotriene B4 is involved in the pathogenesis of postischaemic cerebral oedema. We set out to determine if a relationship exists between jugular bulb leukotriene B4 and other indicators of cerebral ischaemia. No association could be demonstrated for either intracranial pressure, jugular bulb blood oxygen saturation or lactate oxygen index. Our results provide insufficient evidence to suggest that measurement of leukotriene B4 has any place in the management of head-injured patients.


Strategies for Inhibition of Biofilm Formation on Silicone Rubber Voice Prostheses: A Systematic Review.
Lifetime elongation of the silicone voice rubber prostheses by inhibition of biofilm formation is a primary objective in voice restoration of laryngectomized patients. This systematic review sought to explore the existing strategies in this direction. We conducted a systematic search of both in vitro and in vivo literature published in PubMed, Scopus, and Cochrane Central Register of Controlled Trials, until December 31, 2022, for published and unpublished trials assessing the strategies for inhibiting biofilm formation on silicone rubber voice prostheses, and appraised quality assessment with the modified Consolidated Standards of Reporting Trials tool. We analyzed the infection prevention capacity of the included antibacterial and antifungal agents. The qualitative synthesis showed that both surface modification methods and prophylactic treatment of silicone rubber voice prostheses present adequate antibiofilm activity. Of note, the majority of the suggested prosthetic surfaces were not chronically exposed to both human fluids and biofilm-forming microorganisms. Various experimental methods provide promising antibiofilm activity and, thus, possible lifespan elongation of silicone rubber voice prostheses.


Purification and biochemical characterization of Escherichia coli RecA proteins mutated in the putative DNA binding site.
Escherichia coli RecA protein plays a central role both in DNA repair and in recombination. We report biochemical properties of three new RecA proteins mutated at positions 199 (RecA694), 207 (RecA659), and 211 (RecA611) in the putative DNA binding site. RecA694 had a wild-type phenotype, whereas RecA611 and RecA659 were deficient in promoting both the self-cleavage of LexA repressor and the DNA-strand exchange reaction. In order to determine the origin of this inhibition, we examined the capacity of wild-type and mutant proteins to bind to single-stranded DNA (with and without single-stranded binding protein, SSB), double-stranded DNA, and ATP. DNA strand exchange defects were correlated with the inability of mutant proteins to displace SSB from DNA. For the recA659 mutation this inhibition was reversed by equimolar wild-type protein. In contrast, mixtures of either wild-type/RecA659 or wild-type/RecA611 proteins remained deficient in LexA cleavage, suggesting that the dominant negative phenotype of the mutant proteins may be a consequence of the formation heterologous RecA complexes. Various mutations in the putative DNA binding site of RecA protein altered ATP binding, ATPase activity, displacement of SSB from single-stranded DNA, and protein-protein interactions. These results are consistent with the hypothesis that DNA binding to this site of RecA relays allosteric effects to several functional domains throughout the protein.


Prevalence of Cardiovascular Risk Factors and Related Medical and Lifestyle Interventions Among Italian Cardiovascular Specialists: A Proof-of-Concept Study.
Physicians and researchers in the cardiovascular field are constantly engaged in the promotion of guidelines-directed preventive measures, but whether they are themselves adherent to the same recommendations was only sporadically examined. To assess awareness of self-exposure to cardiovascular risk factors and related management among cardiovascular specialists. During the National Conference of the Italian Society of Hypertension (October 2022), a pilot observational study on consecutive volunteer cardiovascular specialists was conducted. Participants underwent standard sitting and standing blood pressure (BP) measurements and answered a questionnaire regarding modifiable/non modifiable cardiovascular risk factors and related treatments. Based on self-declarations and actual measurements, BP was classified as optimal, normal, high-normal BP, and new hypertension in untreated participants, and as treated/untreated pre-existing hypertension. Controlled hypertension was defined as BP < 140/90 mmHg; age-adjusted lower targets were also applied, according to guidelines. In total, 62 participants (30 F, mean age 43.2 ± 14.8 years) were enrolled; 79% reported regular physical activity; 53% of women and 38% of men were on a low-salt diet. After smoke (19.4%), dyslipidemia was the second most common risk factor (17.7%), often occurring with high BP (26.3%) and left untreated (36.7%). Pre-existing hypertension (11.3%) was often uncontrolled (57.1%) and associated with non-adherence to guidelines-directed lifestyle recommendations. About one in 12 participants was unaware of having high measured BP values. Despite the specific professional exposure, a margin for improvement in self cardiovascular risk factors awareness and management remains in this exploratory sample of cardiovascular specialists. This pilot research anticipates forthcoming, larger studies during national and international conferences.


Mandible fractures and dental trauma.
Appropriate and timely management of traumatic injury to the teeth and their supporting structures plays an important role in determining long-term outcome. This article reviews the relevant anatomy and describes the initial diagnosis and treatment approach to dentoalveolar traumatic injury and fractures of the mandible. Procedures for initial stabilization and appropriate referral are also discussed.


Outcomes of total hip arthroplasty after failed free vascularized fibular grafting for osteonecrosis of the femoral head.
Total hip arthroplasty (THA) after vascularized fibular grafting (VFG) is technically demanding with poorer outcomes compared to primary THA in patients with osteonecrosis of the femoral head (ONFH). The purpose of this study was to compare the outcomes of THA after VFG after removal of the graft with that of primary THA. ONFH patients treated by VFG at a single institution were studied retrospectively. THAs after VFG performed by single arthroplasty surgeon with a single type of THA prosthesis were enrolled in the study. A control cohort of patients was created by 1:1 matching with the THA after VFG cohort according to age, gender, and American Society of Anesthesiology (ASA) score from ONFH patients treated by primary THA. Early and long-term outcomes were compared between the two groups. A total of 24 hips were included in the THA after VFG group and compared with 24 primary THA hips. No significant difference was noted in stem position. The THA after VFG group had increased blood loss and longer duration of operation time. There were two cases of intraoperative femur fractures in the THA after VFG group. The femoral stem of both groups were stable, with no cases of revision, and similar HHS scores at the last follow-up. THA after VFG may have similar short to midterm outcomes with primary THA after removal of the residual abutting bone graft and correct positioning of the femoral stem.


Adequate iodine levels in healthy pregnant women. A cross-sectional survey of dietary intake in Turkey.
To assess current iodine levels and related factors among healthy pregnant women. In this cross-sectional, hospital-based study, healthy pregnant women (n=135) were scanned for thyroid volume, provided urine samples for urinary iodine concentration and completed a questionnaire including sociodemographic characteristics and dietary habits targeted for iodine consumption at the Department of Obstetrics and Gynecology, School of Medicine,Muğla Sıtkı Koçman University, Muğla, Turkey, between August 2014 and February 2015. Sociodemographic data were analyzed by simple descriptive statistics. Median urinary iodine concentration was 222.0 μg/L, indicating adequate iodine intake during pregnancy. According to World Health Organization (WHO) criteria, 28.1% of subjects had iodine deficiency, 34.1% had adequate iodine intake, 34.8% had more than adequate iodine intake, and 3.0% had excessive iodine intake during pregnancy. Education level, higher monthly income, current employment, consuming iodized salt, and adding salt to food during, or after cooking were associated with higher urinary iodine concentration. Iodine status of healthy pregnant women was adequate, although the percentage of women with more than adequate iodine intake was higher than the reported literature.


A case of death from the explosion of a 66mm M72 High Explosive Anti-Tank rocket.
A 53-year-old male died from an explosion of a 66 mm M72 High Explosive Anti-Tank (HEAT) rocket. He had collected various cast-off military weapons and was selling them. There were numerous explosive injuries on the anterior side of the body, thus especially bilateral hands, left toe and right knee were severely crushed and fractured. The location and severity of the injuries suggest that he was down on his left knee and was manipulating the weapon with both hands at the moment of detonation. We consider that 66 mm M72 HEAT rocket accidentally detonated during his handling. Very rarely are civilians killed by a military weapon, except during wartime. Appropriate investigation of various explosive injuries provide not only evidence of the cause of death, but also the position and posture of the body.


Cough headache secondary to spontaneous intracranial hypotension complicated by cerebral venous thrombosis.
Cough headache may be the clinical manifestation, sometimes isolated, of an intracranial disease. There are several possible causes of secondary cough headache. The hypothesis that cough headache may be the expression of spontaneous intracranial hypotension has been advanced only recently. In fact, this would represent an exception to the rule that cough headache is generally secondary to conditions leading to an increase in intracranial pressure and/or volume. We report and discuss a case of cough headache secondary to spontaneous intracranial hypotension in an otherwise healthy 59-year-old man. The condition was complicated by cerebral venous thrombosis.


Gibberellin-induced formation of tension wood in angiosperm trees.
After gibberellin had been applied to the vertical stems of four species of angiosperm trees for approximately 2 months, we observed eccentric radial growth that was due to the enhanced growth rings on the sides of stems to which gibberellin had been applied. Moreover, the application of gibberellin resulted in the formation of wood fibers in which the thickness of inner layers of cell walls was enhanced. These thickened inner layers of cell walls were unlignified or only slightly lignified. In addition, cellulose microfibrils on the innermost surface of these thickened inner layers of cell walls were oriented parallel or nearly parallel to the longitudinal axis of the fibers. Such thickened inner layers of cell walls had features similar to those of gelatinous layers in the wood fibers of tension wood, which are referred to as gelatinous fibers. Our anatomical and histochemical investigations indicate that the application of gibberellin can induce the formation of tension wood on vertical stems of angiosperm trees in the absence of gravitational stimulus.


Perioperative pain management.
The management of perioperative pain starts with the use of approaches to minimize anxiety and distress before the procedure. The administration of analgesics or local anesthetics before the start of surgery reduces the nociceptive input occurring during the procedure and reduces the need for postoperative analgesics. As the animal recovers from anesthesia, it is important to administer analgesics to minimize the patient's experience of pain and to continue this therapy through at least the first 12 to 24 hours. Techniques that provide a continuous level of analgesia are more effective than those that allow the pain to return.


Prostaglandin analog effects on cerebrospinal fluid reabsorption via nasal mucosa.
Cerebrospinal fluid (CSF) outflow has been demonstrated along nasal lymphatics via olfactory nerve projections; flow may be increased by stimulating lymphatic contractility using agents such as noradrenaline and the thromboxane A2 analog U46619. Lymphatics elsewhere in the body show increased contractility upon exposure to the prostaglandin F2alpha analog isoprostane-8-epi-prostaglandin. We investigated the ability of ophthalmic prostaglandin F2alpha analogs to increase CSF outflow when applied to the nasal mucosa by inhalation. Latanoprost (0.1, 0.5, or 1mg/ml), bimatoprost (0.3 or 3mg/ml), travoprost (0.04 or 0.4mg/ml), latanoprostene bunod (0.24 or 2.4mg/ml), tafluprost (0.25 or 2.5mg/ml), or control vehicle (10% DMSO) was administered to awake adult C57B/6 mice by nasal inhalation of 2μl droplets. Multiday dosing (daily for 3 days) of latanoprost also was evaluated. A total of 81 animals were studied including controls. General anesthesia was induced by injection, and fluorescent tracer (AlexaFluor647-labelled ovalbumin) was injected under stereotaxic guidance into the right lateral ventricle. Nasal turbinate tissue was harvested and homogenized after 1 hour for tracer detection by ELISA and fluorometric analysis. Inhalation of latanoprost 0.5mg/ml and 1mg/ml led to a 11.5-fold increase in tracer recovery from nasal turbinate tissues compared to controls (3312 pg/ml vs 288 pg/ml, p<0.001 for 0.5mg/ml; 3355 pg/ml vs 288 pg/ml, p<0.001 for 1mg/ml), while latanoprost 0.1 mg/ml enhanced recovery 6-fold (1713 pg/ml vs 288 pg/ml, p<0.01). Tafluprost 0.25mg/ml and bimatoprost 0.3mg/ml showed a modest (1.4x, p<0.05) effect, and the remaining agents showed no significant effect on tracer recovery. After 3 days of daily latanoprost treatment and several hours after the last dose, a persistently increased recovery of tracer was found. Prostaglandin F2alpha analogs delivered by nasal inhalation resulted in increased nasal recovery of a CSF fluorescent tracer, implying increased CSF outflow via the nasal lymphatics. The greatest effect, partially dose-dependent, was observed using latanoprost. Further studies are needed to determine the efficacy of these agents in reducing ICP in short and long-term applications.


Sensitive and selective liquid chromatographic postcolumn reaction detection system for biotin and biocytin using a homogeneous fluorophore-linked assay.
A homogeneous fluorophore-linked assay was used to develop a postcolumn reaction detection system for high-performance liquid chromatography (HPLC). Biotin and biocytin were chosen as the model analytes. The effluent from the HPLC column was merged with a reagent stream containing avidin that was labeled with fluorescein isothiocyanate (avidin-FITC). The binding of the separated analytes by the labeled avidin was accompanied by an enhancement of the fluorescence intensity at 520 nm. This increase in fluorescence was proportional to the concentration of the analytes and constituted the analytical signal. The procedure was optimized with respect to the reagent concentration and the flow-rate of the reagent solution. Analytical characteristics of the method were determined. The procedure was highly selective for biotin and its derivatives. The detection limits for biotin and biocytin were 89 and 94 pg, respectively, for 20-microliters injections. The developed postcolumn reaction detection system was validated by determining biotin in a liquid vitamin preparation and a horse-feed supplement.


[Structure-activity relationship in reactivators of acetylcholinesterase inhibited by phosphoric esters. XII. Hydroxyiminomethylalkylpyridine iodomethylate].
2-Hydroxyiminomethyl-4-alkylpyridine methiodides were prepared and tested for their in vitro reactivating potency on phosphorylated electric eel cholinesterase. They were also tested as in vivo protecting agents: only one of the compounds, 4-methyl-2-hydroxyiminopyridine, offered a greater protection from DFP than 2-PAM, the others had little to no effect.


Enhancement of esculetin on Taxol-induced apoptosis in human hepatoma HepG2 cells.
The potential use of low dose chemotherapy has been appealing since lower dosages are more attainable during cancer therapy and cause less toxicity in patients. Combination therapy of Taxol, a promising frontline chemotherapy agent, with natural anti-tumor agents that are considerably less toxic with a capability of activating additional apoptotic signals or inhibiting survival signals may provide a rational molecular basis for novel chemotherapeutic strategies. Esculetin, a well-known lipoxygenase inhibitor, showed an inhibitory effect on the cell cycle progression of HL-60 cells in our previous study. In this report, the effects of a concomitant administration of esculetin and Taxol were investigated in human hepatoma HepG2 cells. Firstly, esculetin alone could exert an antiproliferation effect together with an inhibitory effect on the activation of ERKs and p38 MAPK. As compared to the treatment with Taxol only, a co-administration with esculetin and Taxol could result in a further enhancement of apoptosis as revealed by DNA fragmentation assay and Annexin-V-based assay. Meanwhile, immunoblotting analysis also showed that the co-administration of esculetin and Taxol could increase the expression of Bax and the cytosolic release of cytochrome C and enhance the expression of Fas and Fas ligand while the activation of caspase-8 and caspase-3 was also increased. Finally, the ERK cascade was proven to be involved in the enhancement of esculetin on the Taxol-induced apoptosis.


Diagnosis and treatment of talar dislocation fractures illustrated by 3 case reports and review of literature.
Talar fractures are a rare type of fractures (less than 1%). They are difficult to treat and outcome is often complicated by arthritis and avascular necrosis. In this article three cases are presented with different types of dislocated talar neck fractures. Anatomy of the talus, treatment, outcome and follow up of these fractures are discussed. Further, review of literature and guidelines for treatment and follow up for dislocated talar neck fractures are discussed. The risk of developing arthritis or avascular necrosis of the talus after dislocated talar neck fractures depends on the initial trauma with vascular compromise due to dislocation of the talus. The modified Hawkins classification gives an insight in the risk of developing avascular necrosis. During follow up the Hawkins sign can be an indication of a vital talus. To diagnose avascular necrosis MRI is the only suitable diagnostic tool. Reduction of a dislocated talar fracture is a medical emergency in an effort to reduce the vascular compromise of the talus. Definitive fixation can be delayed but should be performed by an experienced surgeon to achieve an optimal reconstruction of the talar surface. Long-term follow up is important to evaluate signs of arthritis and avascular necrosis.


Immunohistochemical Expression of Nanog and Its Relation with Clinicopathologic Characteristics in Breast Ductal Carcinoma
Cancer stem cells (CSCs) are a group of tumor cells with self-renewal property and differentiation potential. CSCs play a crucial role in malignant progression of several types of tumors. However, what is still controversial is the clinicopathological relationship between the Nanog marker and its prognostic value in the patients with breast cancer. The expression of Nanog in the patients with breast cancer and its correlation with clinicopathological prognostic factors was explored in the present study. A sample of 120 breast cancer tissues was obtained from the patients who referred to Imam Khomeini Hospital in Sari City, Iran during January 2012 and December 2016. The associations between Nanog expression and clinicopathological factors were analyzed based on immunohistochemical analysis. The expression of Nanog was detected in 67 (55.8%) patients with a high expression rate in 24 (36%) cases (staining index ≥3). Moreover, there was a statistically significant relationship between Nanog expression and clinicopathological factors, including tumor grade (p = 0.001), lymph node metastasis (p = 0.01), and the stage of the disease (p = 0.003). Findings of the study indicate that Nanog may act as a biomarker for prognostic prediction in patients with breast cancer.


Optimizing imaging after coarctation stenting: the clinical utility of multidetector computer tomography.
Following stenting for coarctation of the aorta, it can be difficult to image the stent adequately using cross-sectional echocardiography. Unless there is marked clinical concern regarding stent patency, repeat cardiac catheterization is not warranted. Multidetector computed tomography (MDCT) is a useful alternative imaging modality. We report a case that highlights the clinical utility of MDCT for imaging patients who have had stenting of aortic coarctation. MDCT clearly revealed stent fracture as the cause of late stent failure and recoarctation.


[Recurrent infective endocarditis. A case report].
We report the case of a 63-year old patient presenting five infective endocarditis (IE) in a 13-years period, with different types of streptococcus. No entrance way had been found until the last relapse occurring on mitro-aortic bioprosthesis. This recurrence leads to the diagnosis of caecal adenocarcinoma by coloscopy, preceded and guided by PET scanner. It seems to be the initial entrance way which had been unrecognized because of its small size and its particular location, that is very difficult to observe by coloscopy. Recurrent streptococcus IE are rare and severe. When a gut or colic tumor is suspected, PET scan seems to be helpful in etiological survey.


Hyperperfusion and enhancement in dynamic computed tomography of ischemic stroke patients.
The passage of contrast medium was observed using serial computed tomography (CT) in 24 stroke patients. Density--time profiles of various brain regions were plotted. In normal brain tissue, X-ray attenuation showed a maximum increase during the arterial phase (16.4 +/- 11.0%) and was 2.8 +/- 2.2% above control during stable distribution. In hypoperfusion, increase in attenuation was always below 10% in the arterial phase, while hyperperfusion was characterized by an attenuation increase of 25 to 70%. Enhancement was defined by a density increase of 16.8 +/- 14.8% and a tissue/blood ratio between 7 and 60%. An attempt was made to establish a relationship between the serial CT pattern and the prognosis. Enhancement tended to indicate severe morphological changes followed by permanent neurological deficit, whereas hyperperfusion was generally an indicator of probably recovery.


Characterization and expression of MHC class II alpha and II beta genes in mangrove red snapper (Lutjanus argentimaculatus).
The major histocompatibility complex (MHC) class II plays a key role in adaptive immunity by presenting foreign peptides to CD4(+) T cells and by triggering the adaptive immune response. While the structure and function of MHC class II have been well characterized in mammalian, limited research has been done on fishes. In this study, we characterized the gene structure and expression of MHC class II α (Lunar-DAA) and II β (Lunar-DAB) of mangrove red snapper (Lutjanus argentimaculatus). Both genes shared, respectively, a high similarity and typical features with other vertebrate MHC class II α and II β. The phylogenetic analysis of the deduced peptides revealed that both Lunar-DAA and Lunar-DAB were located in the teleost subclass. Western blotting analyses indicated that both MHC class II α and II β were expressed ubiquitously in immune-related cells, tissues and organs, and that MHC class II α and II β chains existed mainly as heterodimers. While it was highly expressed in gills, thymus, head kidney (HK), spleen, head kidney macrophage and spleen leucocytes, MHC class II β chain was expressed with a low abundance in skin, intestine, stomach and heart. The highest expression of MHC class II β in thymus confirmed the conclusion that thymus is one of the primary lymphoid organs in fishes. The detection of MHC class II αβ dimers in HK macrophages and spleen leucocytes indicated that HK macrophages and spleen leucocytes play a critical role in the adaptive immunity in fishes. All these results provide valuable information for understanding the structure of MHC class II α and II β and their function in immune responses.


Epstein‒Barr virus-associated cellular immunotherapy.
Epstein‒Barr virus (EBV) is a human herpes virus that is saliva-transmissible and universally asymptomatic. It has been confirmed that more than 90% of the population is latently infected with EBV for life. EBV can cause a variety of related cancers, such as nasopharyngeal carcinoma, diffuse large B-cell lymphoma, and Burkitt lymphoma. Currently, many clinical studies have demonstrated that EBV-specific cytotoxic T lymphocytes and other cell therapies can be safely and effectively transfused to prevent and treat some diseases caused by EBV. This review will mainly focus on discussing EBV-specific cytotoxic T lymphocytes and will touch on therapeutic EBV vaccines and chimeric antigen receptor T-cell therapy briefly.


Associated predictors of functional impairment among adolescents with ADHD-a cross-sectional study.
Attention-deficit/hyperactivity disorder (ADHD) in adolescence is associated with functional impairment in several domains of life. To enable development of interventions that more effectively target functional impairment in this age group, the associations between clinical characteristics and impairment need to be clarified. The aim of this study was to investigate the associations between ADHD and functional impairment, if they varied by sex, and the potential impact of comorbid psychiatric symptoms on the associations. This was a cross-sectional study including adolescents with ADHD (n = 164) and a reference group of adolescents without ADHD (n = 106). Self-ratings and parental ratings of functional impairment in different life domains were used as outcomes in all analyses. Differences between groups were investigated with comparative analyses. General linear models (GLMs) were used to explore associations between ADHD symptoms and functional impairment in adolescents with ADHD, while adjusting for of comorbid symptoms, sex, and medication. Adolescents with ADHD displayed higher levels of functional impairment than peers without ADHD, and girls with ADHD rated higher impairment than their male counterparts. The combined ADHD presentation was associated with the highest levels of self-reported impairment, while parental ratings indicated comparable levels of overall impairment across presentations. In the adjusted GLMs, symptoms of inattention were strongly associated with self- and parent-rated impairment in school, but symptoms of hyperactivity/impulsivity were not, whereas symptoms of both inattention and hyperactivity/impulsivity were modestly associated with self-rated impairment with friends. Further, both emotional and conduct problems were associated with impairment in daily life. Our results suggest that attention difficulties, in particular, seem to impair academic functioning in adolescents with ADHD, and interventions targeting such difficulties are warranted. In addition, comorbid symptoms need to be assessed and treated, and self-reports of functioning should be included in research and clinical practice involving adolescents.


Microbial diversity and activity along the forefields of two receding glaciers.
Forefields of two receding glaciers were sampled along either a 150 or 200 m long transect at identical spatial intervals for assessment of soil microbial activity and community diversity trends. The forefields belonged to the Dammaglacier (forefield area is 157 ha, 2000 m above sea level) and Rotfirnglacier (100 ha, 2200 m) and at the time of sampling were receding at an estimated rate of 8 and 10 m yr(-1) over the past 5 years, respectively. Direct counting of bacteria (DAPI staining), assessment of dehydrogenase activity (DH), and fluorescein diacetate hydrolysis activity (FDA) were performed to estimate bacteria number and soil microbial activity. Along the Dammaglacier forefield (from youngest to oldest soil), bacteria number (8.21 x 10(7) to 1.49 x 10(9) cells g(-1) soil), DH activity (0 to 61 mg TTC reduced g(-1) soil h(-1)), and FDA activity (0 to 100 mg fluorescein produced g-1 soil h-1) increased, suggesting the development of microbial populations increasing in number and activity. The Rotfirn forefield exhibited similar trends per gram of soil in bacteria number (1.13 x 10(8) to 5.93 x 10(9) cells), DH activity (0 to 36 mg TTC reduced), and FDA activity (2 to 70 mg fluorescein produced), but with more variability among samples than the Damma forefield samples. Molecular assessment of bacterial diversity included denaturing gradient gel electrophoresis (DGGE) and ribosomal intergenic spacer analysis (RISA) of soil DNA. DGGE and RISA revealed that the composition and succession of bacterial populations were different in both forefields. Comparison of Shannon diversity index values indicated that all populations sampled from the Damma forefield were significantly different (p < 0.05). Conversely, similar populations existed in the Rotfirn forefield succession. Overall, the results indicate that diverse bacterial assemblages increasing in number and activity characterize these glacier forefield soils with both forefield successions exhibiting differing modes of bacterial community establishment.


Novel chitosan film embedded with liposome-encapsulated phage for biocontrol of Escherichia coli O157:H7 in beef.
In recent years, phages used for the reduction of pathogenic bacteria have fostered many attentions, but they are liable to lost bioactivity in food due to the presence of acidic compounds, enzymes and evaporite materials. To improve the stability of phages, a chitosan edible film containing liposome-encapsulated phage was engineered in the present study. The characteristics of liposome-encapsulated phage and the chitosan film containing liposome-encapsulated phage were investigated. The encapsulation efficiency of phages in liposome reached 57.66±0.12%. Besides, the desirable physical properties of chitosan film were obtained. The chitosan film embedded with liposome-encapsulated phage exhibited high antibacterial activity against Escherichia coli O157:H7, without the impact on the sensory properties of beef. Hence, chitosan film containing liposome-encapsulated phage could be a promising antibacterial packaging for beef preservation.


Interactions of lanthanum with purified and intact cell acetylcholinesterase of Electrophorus electricus.
The effects of lanthanum on the activity of purified preparations of acetylcholinesterase (AChE) from the electric organ of E. electricus and on the activity of AChE in intact electroplaques from the same species were studied. 0.1 mM LaCl3 produced an initial inhibition of purified AChE which was followed by a delayed activation of the enzyme. Upon pretreatment of purified enzyme with LaCl3, initial activity was markedly increased. LaCl3 exerted a marked, concentration-dependent inhibition of intact cell AChE. La3+ and Ca2+ appear to interact competitively. In the presence of both 10 mM CaCl2 and 0.1 mM LaCl3, the initial activitity of purified AChE was increased at lower ACh concentrations and inhibited at ACh concentrations greater than 3 X 10(-4) M. Inhibition of intact cell enzyme by 0.1 mM LaCl3 was relieved by increasing the CaCl2 concentration to 10 mM at ACh concentrations less than 2 X 10(-4) M. The data were analyzed assuming Michaelis-Menten kinetics and interpreted with reference to the differential binding of divalent and trivalent cations to regulatory anionic sites which are separate and distinct from the anionic site of the active center of the enzyme.


Development and Testing of a Novel Locking Pin Cap to Create a Fixed-Angle K-Wire Plate Construct.
To test the effectiveness of a novel locking pin cap to attach a K-wire rigidly to a volar locking plate and resist fracture displacement compared with commercially available alternatives. Two different methods of fracture fixation were tested on a total of 12 Sawbones models with volar shear distal radius fracture (6/group). The fragments were fixed with either 2 commercially available pin plates (industry standard) or a volar plate with 2 locking screws fixing the scaphoid facet and 2 pins locked to the plate with a novel locking pin cap in the lunate facet. Axial load conditioning was performed followed by sinusoidal loading to 250 N at 50 mm/s. A motion capture system was used to assess the relative movement of the fracture fragments relative to the intact shaft. The strength of the fixation construct was quantified by (1) the force required to achieve a 2-mm gap between the shaft and fracture fragments and (2) ultimate load to failure. One industry standard pin plate demonstrated disassociation of the pin from the plate after fatigue conditioning. This did not occur in the locking pin cap group. The locking pin cap construct group was able to sustain a significantly higher load compared with the industry standard when the construct was displaced to the 2-mm gap. The locking pin cap also significantly increased the ultimate load to failure compared with the industry standard. The novel locking pin cap creates a fixed-angle attachment of a K-wire to an existing locking screw hole in a plate. This fixed-angle K-wire is significantly stronger in preventing gap formation and resisting ultimate failure than commercially available plates that use bent K-wires. The development of novel techniques to secure small articular fragments may ultimately improve clinical outcomes.


Operant conditioning of the primate H-reflex: factors affecting the magnitude of change.
Primates can gradually increase or decrease H-reflex amplitude in one leg when reward depends on that amplitude. The magnitude of change varies greatly from animal to animal. This study sought to define the factors that control this magnitude. It evaluated the influence of animal age, muscle size (absolute and relative), background electromyographic activity (EMG) level, M response amplitude, initial H-reflex amplitude, performance intensity, and behavior of the contralateral leg. Fifty-four animals (Macaca nemestrina) underwent operant conditioning of the triceps surae H-reflex in one leg (the trained leg). Twenty-eight were rewarded for larger H-reflexes (HRup animals), and 26 were rewarded for smaller H-reflexes (HRdown animals). In the HRup animals, H-reflex amplitude in the trained leg rose to an average final value of 177% of its initial amplitude. Magnitude of increase varied widely across animals. Nine animals rose to 120-140%, 11 to 160-240%, three to 300% or more, and five remained within 20% of initial amplitude. In the HRdown animals, H-reflex amplitude in the trained leg decreased to an average of 69% of initial amplitude. Magnitude of decrease varied widely. Five animals decreased to 20-40%, seven to 40-60%, six to 60-80%, and eight remained within 20% of initial amplitude. Animal age, as assessed by weight, markedly affected HRdown conditioning, but not HRup conditioning. Heavy HRdown animals (> or = 6 kg) were more successful than light HRdown animals (< 6 kg). Thirteen of 14 heavy animals and only five of 12 light animals decreased to less than 80% of initial amplitude.(ABSTRACT TRUNCATED AT 250 WORDS)


[Frequency of fused roots, divergent separation, convergent separation, rectilinear separation and delaceration of third molars].
In a sample of Brazilian white young adults between 18-23 years old, the A.A. studied the third molar roots, in anatomical aspects, using the orthopantomographic radiographic method. They concluded that the superior third molar shows a biggest number of fusionated roots (19.50% in the right side and 19.66% in the left side) and the inferior third molar shows a biggest number of separated roots (21.48% in the right side and 19.66% in the left side).


CircUSP48 promotes malignant behavior by regulating CYR61 via miR-365 in osteosarcoma.
Even though circular RNAs (circRNAs), a class of non-coding endogenous RNA, play a crucial role in the progression of osteosarcoma (OS), the specific function of hsa_circ_0000028 (circUSP48) remains unclear. This study aims to elucidate the mechanism by which circUSP48 regulates OS. We employed qRT-PCR and western blot techniques to quantify circDOCK1, miR-186, and DNMT3A levels. Cell proliferation was assessed using the cell counting kit-8 (CCK-8), 5-Ethynyl-20-deoxyuridine (EdU) assay, and colony formation assay. Cell migration and invasion were evaluated through Transwell and cell scratch assays. Furthermore, we performed dual-luciferase reporter, RIP, and RNA pull-down assays to investigate the association between circUSP48, miR-365, and CYR61. In addition, an in vivo xenograft model was utilized to assess the functional role of circUSP48. High levels of circUSP48 and CYR61 were observed in OS tissues and cells, while miR-365 levels were low. Knockdown of circUSP48 suppressed the multiplication, motility, and invasion of OS cells, thereby reducing carcinoma growth. Moreover, inhibition of miR-365 reversed the OS cell-suppressive effect caused by circUSP48 knockdown through direct interaction with circUSP48. Additionally, circUSP48 upregulated the expression of CYR61 by sponging miR-365. The findings suggest that circUSP48 promotes malignant behavior in OS by regulating the expression of CYR61 through miR-365, making it a potential therapeutic target for OS.


Structure of Leishmania donovani coronin coiled coil domain reveals an antiparallel 4 helix bundle with inherent asymmetry.
Coiled coils are ubiquitous structural motifs that serve as a platform for protein-protein interactions and play a central role in myriad physiological processes. Though the formation of a coiled coil requires only the presence of suitably spaced hydrophobic residues, sequence specificities have also been associated with specific oligomeric states. RhXXhE is one such sequence motif, associated with parallel trimers, found in coronins and other proteins. Coronin, present in all eukaryotes, is an actin-associated protein involved in regulating actin turnover. Most eukaryotic coronins possess the RhXXhE trimerization motif. However, a unique feature of parasitic kinetoplastid coronin is that the positions of R and E are swapped within their coiled coil domain, but were still expected to form trimers. To understand the role of swapped motif in oligomeric specificity, we determined the X-ray crystal structure of Leishmania donovani coronin coiled coil domain (LdCoroCC) at 2.2Å, which surprisingly, reveals an anti-parallel tetramer assembly. Small angle X-ray scattering studies and chemical crosslinking confirm the tetramer in solution and is consistent with the oligomerization observed in the full length protein. Structural analyses reveal that LdCoroCC possesses an inherent asymmetry, in that one of the helices of the bundle is axially shifted with respect to the other three. The analysis also identifies steric reasons that cause this asymmetry. The bundle adapts an extended a-d-e core packing, the e residue being polar (with an exception) which results in a thermostable bundle with polar and apolar interfaces, unlike the existing a-d-e core antiparallel homotetramers with apolar core. Functional implications of the anti-parallel association in kinetoplastids are discussed.


Serrated (W/M) osteotomy: a new technique for simultaneous correction of angular and torsional deformity of the lower limb in children.
The serrated (W/M) osteotomy allows for simultaneous correction of angular and/or torsional deformities of the lower limb in the growing child. This osteotomy provides an attractive alternative to various types of upper tibial or distal femoral osteotomies. It eases the translations of the preoperative clinical and radiological evaluation to accurate angles during surgery. This type of osteotomy is stable, with large contact surfaces that provide excellent healing. Although no length is sacrificed, neither bone graft nor internal or external fixation is required.