Utility of machine learning in developing a predictive model for early-age-onset colorectal neoplasia using electronic health records.
The incidence of colorectal cancer (CRC) is increasing in adults younger than 50, and early screening remains challenging due to cost and under-utilization. To identify individuals aged 35-50 years who may benefit from early screening, we developed a prediction model using machine learning and electronic health record (EHR)-derived factors. We enrolled 3,116 adults aged 35-50 at average-risk for CRC and underwent colonoscopy between 2017-2020 at a single center. Prediction outcomes were (1) CRC and (2) CRC or high-risk polyps. We derived our predictors from EHRs (e.g., demographics, obesity, laboratory values, medications, and zip code-derived factors). We constructed four machine learning-based models using a training set (random sample of 70% of participants): regularized discriminant analysis, random forest, neural network, and gradient boosting decision tree. In the testing set (remaining 30% of participants), we measured predictive performance by comparing C-statistics to a reference model (logistic regression). The study sample was 55.1% female, 32.8% non-white, and included 16 (0.05%) CRC cases and 478 (15.3%) cases of CRC or high-risk polyps. All machine learning models predicted CRC with higher discriminative ability compared to the reference model [e.g., C-statistics (95%CI); neural network: 0.75 (0.48-1.00) vs. reference: 0.43 (0.18-0.67); P = 0.07] Furthermore, all machine learning approaches, except for gradient boosting, predicted CRC or high-risk polyps significantly better than the reference model [e.g., C-statistics (95%CI); regularized discriminant analysis: 0.64 (0.59-0.69) vs. reference: 0.55 (0.50-0.59); P<0.0015]. The most important predictive variables in the regularized discriminant analysis model for CRC or high-risk polyps were income per zip code, the colonoscopy indication, and body mass index quartiles. Machine learning can predict CRC risk in adults aged 35-50 using EHR with improved discrimination. Further development of our model is needed, followed by validation in a primary-care setting, before clinical application.


Genome-wide mapping for fatty acid composition and melting point of fat in a purebred Duroc pig population.
The fatty acid composition and melting point of fatty tissue are among the most important economic traits in pig breeding because of their influence on the eating quality of meat. Identifying the quantitative trait locus (QTL) of these traits may help reveal the genetic structure of fatty acid composition and the melting point of fatty tissue and improve meat-quality traits by marker-assisted selection. We conducted whole-genome QTL analysis for fatty acid composition and melting point of inner and outer subcutaneous fat and inter- and intramuscular fat in a purebred Duroc population. A total of 129 markers were genotyped and used for QTL analysis. For fatty acid compositions of inner and outer subcutaneous fat, three significant QTL and 17 suggestive QTL were detected on SSC2, 4, 6, 8, 9, 10, 11, 12, 14 and 18. For the melting point of inner and outer subcutaneous fat, two significant QTL were detected on the same region of SSC14. For fatty acid compositions of inter- and intramuscular fat, five significant QTL and 13 suggestive QTL were detected on SSC2, 4, 6, 8, 9, 10, 14 and 15. On SSC14, significant QTL for C18:0 and C18:1 of outer subcutaneous fat and intramuscular fat, and melting point of subcutaneous fat, which had high likelihood of odds (LOD) scores (2.67-5.78), were detected in the same region. This study determined QTL affecting fatty acid composition and melting point of different fat tissues in purebred Duroc pigs.


Total hip arthroplasty using a cylindrical cementless stem in patients with a small physique.
We performed total hip arthroplasty using an anatomic medullary locking cementless stem for small-physique patients from 1988 to 1995. We conducted a retrospective study of 50 joints in 44 cases, including 40 developmentally dysplastic hips followed for 12 to 20 years (average, 15.1 years). Average height and body weight were 152 cm and 56 kg (5.0 ft and 124 lb), respectively, with an average body mass index of 24.2. Twelve joints (24%) were revised for acetabular-sided failures. Forty-eight stems (96%) showed bone ingrowth fixation, and there were no unstable stems. The simple cylindrical shape of the distal portion of the AML stem was less affected by deformity of the proximal femur of developmental dysplasia of the hip in patients with a small physique, and both clinically and radiologically good results were confirmed at long-term follow-up.


Peer-mentees' challenges in an undergraduate peer-group clinical mentoring programme in a nursing education institution.
Clinical competency and professional growth amongst nursing students is the cornerstone of a nursing education programme. The demanding and complex training of nursing students requires various clinical teaching and learning strategies such as peer-group clinical mentoring. The objective of this research was to explore and describe the challenges that peer-mentees experience in an undergraduate peer-group clinical mentoring programme in one-specific nursing education institution in the North West Province. The study was conducted at a nursing education institution in North West Province. A qualitative, retrospective case study research design was used. Two separate World Café sessions following a semi-structured schedule based on Gibbs Reflective Cycle were conducted with 51 peer-mentees who were mentored in clinical practice. Four levels of qualitative thematic data analyses were employed to analyse the data. Five themes emerged from the findings of the study, namely, poor implementation of the peer-group clinical mentoring programme, ineffective undergraduate peer-group clinical mentoring programme, undesirable attitudes of the mentors, mentors unprofessional conduct as well as communication challenges. Ten sub-themes emerged from the findings. Literature control was done to support the findings. The findings showed that undergraduate nursing students faced a diversity of challenges in the effective learning and teaching of peer-mentees in a clinical context. Limitations and recommendations of the study were given. Recommendations were given for nursing practice, education and research.


Sequential Events in the Photoinhibition of Synechocystis under Sodium Stress.
Synechocystis (PCC 6714) is photoinhibited at low photon flux density when illuminated in medium devoid of Na(+). Photosynthetic activity and variable fluorescence yield were measured in intact cells during the course of inhibition and during recovery from inhibition in order to characterize sequential inhibitory events. Photosystem II antenna chlorophyll becomes functionally disconnected from the reaction center in the first stage of inhibition, although energy is still transferred from phycobilin pigments to the reaction center. This occurs simultaneously with, or subsequent to, the previously observed (J Zhao, JJ Brand [1988] Arch Biochem Biophys 264: 657-664) inactivation of O(2)-evolution units in Na(+)-stressed cells. Addition of Na(+) to the culture medium rapidly restores the O(2)-evolving system, but reassociation of photosystem II antenna chlorophyll is much slower and requires new protein synthesis. A site nearer to the reaction center of photosystem II becomes inactivated in a secondary inhibitory event. Recovery from secondary inhibition occurs slowly and requires protein synthesis. Cells recovering from secondary inhibition produce active photosystem II units with a fully functional complement of antenna chlorophyll. Synechocystis is photoinhibited in complete (Na(+)-containing) medium when illuminated at high photon flux density. These photoinhibited cells exhibit the same recovery behavior as cells at the second stage of inhibition in Na(+)-deficient medium.


Errors in imaging the pregnant patient with acute abdomen.
Pregnant women with an acute abdomen present a critical issue due to the necessity for an immediate diagnosis and treatment; in fact, a diagnostic delay could worsen the outcome for both the mother and the fetus. There is evidence that emergencies during pregnancy are subject to mismanagement; however, the percentage of errors in the diagnosis of emergencies in pregnancy has not been studied in depth. The purpose of this article is to review the most common imaging error emergencies. The topics covered are divided into gynecological and non-gynecological entities and, for each pathology, possible errors have been dealt with in the diagnostic pathway, the possible technical errors in the exam execution, and finally the possible errors in the interpretation of the images. These last two entities are often connected owing to a substandard examination, which can cause errors in the interpretation. Consequently, the systemization of errors reduces the possibility of reoccurrences in the future by providing a valid approach in helping to learn from these errors.


Transmission beam pattern and echolocation signals of a harbor porpoise (Phocoena phocoena).
The transmission beam pattern of an echolocating harbor porpoise (Phocoena phocoena) was measured in both the vertical and horizontal planes. An array of seven Brüel and Kjaer 8103 hydrophones connected to an amplifier-line driver module was used to measure the beam patterns. The porpoise was trained to station in a hoop and echolocate a cylindrical target located at a range between 7 and 9 m while the array was located 2 m in front of the hoop. The 3-dB beamwidth in both the vertical and horizontal planes was the same at approximately 16 degrees and the beam was pointed toward the forward direction. The individual hydrophones in both the vertical and horizontal arrays measured signal waveforms that were similar throughout the 40-degree span of the array. The porpoise emitted signals with intervals that were 20 to 35 ms longer than the round trip travel time between the animal and the target. The average source level, peak frequency, and bandwidth were 157 dB, 128 kHz, and 16 kHz, respectively.


Carbonyls as Latent Alkyl Carbanions for Conjugate Additions.
Conjugate addition of carbon nucleophiles to electron-deficient olefins is one of the most powerful methods for forming carbon-carbon bonds. Despite great achievements in controlling the selectivity, variation of the carbon nucleophiles remains largely underexplored, with this approach relying mostly on organometallic reagents. Herein, we report that naturally abundant carbonyls can act as latent carbon nucleophiles for conjugate additions through a ruthenium-catalyzed process, with water and nitrogen as innocuous byproducts. The key to our success is homogeneous ruthenium(II) catalysis, combined with phosphines as spectator ligands and hydrazine as the reducing agent. This chemistry allows the incorporation of highly functionalized alkyl fragments into a vast array of electron-deficient olefins under mild reaction conditions in a reaction complementary to the classical organometallic-reagent-based conjugate additions mediated or catalyzed by "soft" transition metals.


Trial of labor vs. elective repeat cesarean section. AAFP Task Force on Clinical Policies for Patient Care.
A trial of labor after a previous low transverse cesarean section in the absence of ongoing contraindications is safe for most women. A comprehensive review and meta-analysis of the literature was conducted to determine outcomes, costs and women's preferences by method of delivery. About seven of 10 women who undergo a trial of labor after previous low transverse cesarean section can expect to deliver vaginally. The following differences in absolute risks were identified: a trial of labor was associated with a slightly increased risk of uterine rupture (0.24 percent) and a decreased risk of infection and fever (5.2 percent) and postpartum bleeding (0.59 percent) as compared with an elective repeat cesarean section. The difference in risk for newborns with an Apgar score of less than 7 at five minutes was 0.85 percent for infants delivered vaginally after a trial of labor versus infants delivered by elective repeat cesarean section. Financial cost (as measured by hospital charges) of cesarean delivery was 1.66 to 2.4 times greater than the cost of vaginal birth. This difference was due almost entirely to the longer length of hospital stay for women who had a cesarean delivery. When given the opportunity, about two-thirds of eligible women choose a trial of labor over elective repeat cesarean section. The reasons underlying women's preferences for a trial of labor or elective repeat cesarean section were diverse and changed during pregnancy. As a consequence, women's preferences for the method of delivery must be explored and respected throughout pregnancy and during the delivery process.(ABSTRACT TRUNCATED AT 250 WORDS)


Avoiding transmitting identified mutations to offspring using preimplantation genetic diagnosis.
Preimplantation genetic diagnosis has been used to decrease or avoid the risk of transmitting identified mutations to offspring. A 29-year-old woman with spondyloepiphyseal dysplasia congenita and her 30-year-old husband with Marfan syndrome underwent in vitro fertilization with preimplantation genetic diagnosis. Two mutation-negative embryos were transferred into a gestational carrier, who became pregnant with twins and delivered two clinically normal neonates. Statistically, this couple would be predicted to have a 75% chance of producing an affected embryo. Using preimplantation genetic diagnosis, two dually unaffected embryos were selected and transferred. This experience expands the use of preimplantation genetic diagnosis to cases with multiple autosomal dominant single-gene disorders.


Biological Properties and Absolute Configuration of Flavanones From Calceolaria thyrsiflora Graham.
Flavanones (-)-(2S)-5,4'-dihydroxy-7-methoxyflavanone (1) and (-)-(2S)-5,3',4'-trihydroxy-7-methoxyflavanone (2) were isolated from the extracts of Calceolaria thyrsiflora Graham, an endemic perennial small shrub growing in the central zone of Chile. The absolute configuration of these compounds was resolved by optical rotation experiments and in silico calculations. Three analogs (3, 4, and 5) were synthesized to do structure-activity relationships with the biological assays studied. Biological tests revealed that only flavanone 2 exhibited a moderate inhibitory activity against the methicillin-resistant strain S. aureus MRSA 97-77 (MIC value of 50 µg/ml). In addition, flavanone 2 showed a potent, selective, and competitive inhibition of 5-hLOX, which supports the traditional use of this plant as an anti-inflammatory in diseases of the respiratory tract. Also, 2 exhibited cytotoxic and selective effects against B16-F10 (8.07 ± 1.61 µM) but 4.6- and 17-fold lesser activity than etoposide and taxol.


Evaluation of reference genes for quantitative real-time PCR normalization in the scarab beetle Holotrichia oblita.
Quantitative real-time polymerase chain reaction (qPT-PCR) is commonly used to analyze gene expression, however, the accuracy of the normalized results is affected by the expression stability of reference genes. Holotrichia oblita (Coleoptera: Scarabaeidae) causes serious damage to crops. Reliable reference genes in H. oblita are needed for qRT-PCR analysis. Therefore, we evaluated 13 reference genes under biotic and abiotic conditions. RefFinder provided a comprehensive stability ranking, and geNorm suggested the optimal number of reference genes for normalization. RPL13a and RPL18 were the most suitable reference genes for developmental stages, tissues, and temperature treatments; RPL13a and RPS3 were the most suitable for pesticide and photoperiod treatments; RPS18 and RPL18 were the most suitable for the two sexes. We validated the normalized results using odorant-binding protein genes as target genes in different tissues. Compared with the selected suitable reference genes, the expression of OBP1 in antennae, abdomen, and wings, and OBP2 in antennae and wings were overestimated due to the instability of ACTb. These results identified several reliable reference genes in H. oblita for normalization, and are valuable for future molecular studies.


Synthesis of Pyrimidine Fused Quinolines by Ligand-Free Copper-Catalyzed Domino Reactions.
Herein, we report two novel methods for the synthesis of pyrimidine fused quinolines using a one-pot C-C and C-N bond forming strategy from the reaction of 6-aminouracils with 2-bromobenzaldehydes or 2-bromobenzyl bromide derivatives in the presence of 10 mol % CuCl2 without using any ligand. The reaction of 2-bromobenzaldehyde or its derivatives with 6-aminouracils in the presence of K2CO3 as base and a catalytic amount of CuCl2 in DMF medium under microwave heating conditions provides corresponding pyrimidine fused quinoline derivatives in good yields within 30 min. Alternatively, pyrimidine fused quinoline derivatives have been synthesized from the reaction of 2-bromobenzyl bromides with 6-aminouracil derivatives in the presence of molecular oxygen, CuCl2 (10 mol %), and K2CO3 as base in DMF under reflux conditions. Structures of all the products were unambiguously confirmed by spectroscopic techniques and by recording single crystal XRD of 3a.


Treatment of tuberculosis in a region with high drug resistance: outcomes, drug resistance amplification and re-infection.
Emerging antituberculosis drug resistance is a serious threat for tuberculosis (TB) control, especially in Eastern European countries. We combined drug susceptibility results and molecular strain typing data with treatment outcome reports to assess the influence of drug resistance on TB treatment outcomes in a prospective cohort of patients from Abkhazia (Georgia). Patients received individualized treatment regimens based on drug susceptibility testing (DST) results. Definitions for antituberculosis drug resistance and treatment outcomes were in line with current WHO recommendations. First and second line DST, and molecular typing were performed in a supranational laboratory for Mycobacterium tuberculosis (MTB) strains from consecutive sputum smear-positive TB patients at baseline and during treatment. At baseline, MTB strains were fully drug-susceptible in 189/326 (58.0%) of patients. Resistance to at least H or R (PDR-TB) and multidrug-resistance (MDR-TB) were found in 69/326 (21.2%) and 68/326 (20.9%) of strains, respectively. Three MDR-TB strains were also extensively resistant (XDR-TB). During treatment, 3/189 (1.6%) fully susceptible patients at baseline were re-infected with a MDR-TB strain and 2/58 (3.4%) PDR-TB patients became MDR-TB due to resistance amplification. 5/47 (10.6%) MDR- patients became XDR-TB during treatment. Treatment success was observed in 161/189 (85.2%), 54/69 (78.3%) and 22/68 (32.3%) of patients with fully drug susceptible, PDR- and MDR-TB, respectively. Development of ofloxacin resistance was significantly associated with a negative treatment outcome. In Abkhazia, a region with high prevalence of drug resistant TB, the use of individualized MDR-TB treatment regimens resulted in poor treatment outcomes and XDR-TB amplification. Nosocomial transmission of MDR-TB emphasizes the importance of infection control in hospitals.


The distribution of consecutive closed birth intervals in females in Uttar Pradesh.
Most studies of closed birth intervals are regarding their variation at specific orders among females. This paper attempts to study the nature of the distributions of consecutive closed birth intervals. Data from the Uttar Pradesh National Family Health Survey 1998-99 (NFHS-2) were analysed. It was found that, under certain assumptions, the postpartum amenorrhoea period and menstruating interval are negatively associated, indicating that socio-cultural factors are affecting the menstruating interval.


• Family credit scheme hailed a success.
Claims for Family Credit, the tax-free benefit for working families with children, have doubled since the start of the Government's publicity drive, Nicholas Scott, Social Security Minister announced last week.


Direct analysis of intact proteins from Escherichia coli colonies by liquid extraction surface analysis mass spectrometry.
Top-down identification of proteins by liquid extraction surface analysis (LESA) mass spectrometry has previously been reported for tissue sections and dried blood spot samples. Here, we present a modified "contact" LESA method for top-down analysis of proteins directly from living bacterial colonies grown in Petri dishes, without any sample pretreatment. It was possible to identify a number of proteins by use of collision-induced dissociation tandem mass spectrometry followed by searches of the data against an E. coli protein database. The proteins identified suggest that the method may provide insight into the bacterial response to environmental conditions. Moreover, the results show that the "contact" LESA approach results in a smaller sampling area than typical LESA, which may have implications for spatial profiling.


Hypoxia reduces the conduction velocity of the excitation along the striated muscles in man.
The conduction velocity of the excitation along the striated muscles in man, hereafter refered to as muscle fiber conduction velocity (MFCV) was used to estimate the muscle functional state under hypoxaemic hypoxia. The averaged MFCV was counted by the method based on the surface measurements (using bipolar "branched" multielectrodes) of the electrically elicited compound action potentials of the brachial biceps muscle. The arterial and venous blood-gas and acid-base parameters were simultaneously measured and the body temperature was controlled intramuscularly. With afebrile patients under normoxaemia or hypoxaemia following brain stroke, MFCV decreased proportionnally to the arterial partial oxygen pressure (PaO2) rate; a strong positive correlation (r = 0.882) between MFCV and PaO2 was found up to PaO2 of 12.0 kPa and MFCV 3.5 m/s. In the case of over PaO2 = 12.0 kPa, the relationship entered into saturation; the further increase PaO2 was not followed by concomitant changes in MFCV. The results show that MFCV-testing is a valuable tool to evaluate the functional state of the striated muscle under hypoxia.


CSS: cluster similarity spectrum integration of single-cell genomics data.
It is a major challenge to integrate single-cell sequencing data across experiments, conditions, batches, time points, and other technical considerations. New computational methods are required that can integrate samples while simultaneously preserving biological information. Here, we propose an unsupervised reference-free data representation, cluster similarity spectrum (CSS), where each cell is represented by its similarities to clusters independently identified across samples. We show that CSS can be used to assess cellular heterogeneity and enable reconstruction of differentiation trajectories from cerebral organoid and other single-cell transcriptomic data, and to integrate data across experimental conditions and human individuals.


Identifying occult maternal malignancies from 1.93 million pregnant women undergoing noninvasive prenatal screening tests.
Multiple chromosomal aneuploidies may be associated with maternal malignancies and can cause failure of noninvasive prenatal screening (NIPS) tests. However, multiple chromosomal aneuploidies show poor specificity and selectivity for diagnosing maternal malignancies. This multicenter retrospective analysis evaluated 639 pregnant women who tested positive for multiple chromosomal aneuploidies on initial NIPS test between January 2016 and December 2017. Women were assessed using genome profiling of copy-number variations, which was translated to cancer risk using a novel bioinformatics algorithm called the cancer detection pipeline (CDP). Sensitivity, specificity, and positive predictive value (PPV) of diagnosing maternal malignancies were compared for multiple chromosomal aneuploidies, the CDP model, and the combination of CDP and plasma tumor markers. Of the 639 subjects, 41 maternal malignant cancer cases were diagnosed. Multiple chromosomal aneuploidies predicted maternal malignancies with a PPV of 7.6%. Application of the CDP algorithm to women with multiple chromosomal aneuploidies allowed 34 of the 41 (83%) cancer cases to be identified, while excluding 422 of 501 (84.2%) of the false positive cases. Combining the CDP with plasma tumor marker testing gave PPV of 75.0%. The CDP algorithm can diagnose occult maternal malignancies with a reasonable PPV in multiple chromosomal aneuploidies-positive pregnant women in NIPS tests. This performance can be further improved by incorporating findings for plasma tumor markers.


Transporter Associated with Antigen Processing 1 Gene Polymorphisms Increase the Susceptibility to Tuberculosis.
Tuberculosis (TB) is known to result from a complex interaction between the host immune response and Mycobacterium infection. The transporter associated with antigen processing (TAP) plays an important role in the processing and presentation pathways for the Mycobacterium tuberculosis (M. tb) antigen. To investigate the possible association of the TAP1 and TAP2 genes with TB. A total of 449 TB patients and 435 control subjects were included in this study, and single nucleotide polymorphisms (SNPs) in the TAP gene, as well as TAP1 and TAP2 alleles, were genotyped. TAP gene association analysis of TB diseases showed that rs41551515-T in the TAP1 gene was significantly associated with susceptibility to TB (P=7.96E-04, OR=4.124, 95% CI: 1.683-10.102), especially pulmonary TB (PTB, P=6.84E-04, OR=4.350, 95% CI: 1.727-10.945), and the combination of rs1057141-T-rs1135216-C in the TAP1 gene significantly increased the risk of TB susceptibility (P=5.51E-05, OR=10.899, 95% CI: 2.555-46.493). Five novel TAP1 alleles were detected in Yunnan Han people, and the allele frequency of TAP1*unknown_3 (rs41555220-rs41549617-rs1057141-rs1135216-rs1057149-rs41551515: C-A-T-C-C-T) was notably increased in all TB patients, including in the PTB and EPTB subgroups, and was significantly associated with the risk of susceptibility to TB. However, no association between the TAP2 gene and TB was found in this study. Host genetic variants of rs41551515-T and the combination rs1057141-T-rs1135216-C, as well as TAP1*unknown_3 may play a critical role in susceptibility to TB disease.


Suppression of autophagy by BCR/ABL.
Imatinib and second generation BCR/ABL tyrosine kinase inhibitors (TKIs) serve now as standard therapies for patients with chronic myelogenous leukemia (CML); however, CML stem cells are intrinsically insensitive to the cell death-inducing effects of TKIs, allowing the persistence of a "reservoir" of BCR/ABL-expressing CML-initiating cells potentially responsible for disease relapse and progression. Although it is still controversial whether the "insensitivity" of CML stem cells to treatment with TKI is due to BCR/ABL-dependent or independent mechanisms, treatment with IM appears to suppress BCR/ABL-dependent signaling in CML stem cells with no adverse effects on their survival. Recent evidence indicates that BCR/ABL suppresses and treatment of CML cells with IM/TKIs induces autophagy, a genetically-regulated process of adaptation to metabolic stress which could allow tumor cells to become metabolically inert enabling their survival under conditions that may mimic growth factor/nutrients deprivation. Based on this hypothesis, TKI-induced autophagy may "antagonize" TKI-induced cell death and inhibition of autophagy may eliminate this survival mechanism by restoring "sensitivity" of CML stem cells to treatment with IM/TKI. Consistent with this, phenotypically and functionally defined CML-enriched stem cells insensitive to treatment with TKI are efficiently eliminated by the combination of TKI and chloroquine, an inhibitor of late stage autophagy. Thus, inhibition of autophagy may improve the potent and specific effects of TKIs by rendering CML stem cells sensitive to these targeted therapies.


TRP channels in human prostate.
This review gives an overview of morphological and functional characteristics in the human prostate. It will focus on the current knowledge about transient receptor potential (TRP) channels expressed in the human prostate, and their putative role in normal physiology and prostate carcinogenesis. Controversial data regarding the expression pattern and the potential impact of TRP channels in prostate function, and their involvement in prostate cancer and other prostate diseases, will be discussed.


Risk stratification of cutaneous melanoma reveals carcinogen metabolism enrichment and immune inhibition in high-risk patients.
Cutaneous melanoma (CM) is the most lethal form of skin cancer. Risk assessment should facilitate stratified surveillance and guide treatment selection. Here, based on the mRNA-seq data from 419 CM patients in the Cancer Genome Atlas (TCGA), we developed a prognostic 21-gene signature to distinguish the outcomes of high- and low-risk patients, which was further validated in two external cohorts. The signature achieved a higher C-index as compared with other known biomarkers and clinical characteristics in both the TCGA and validation cohorts. Notably, in high-risk patients the expression levels of three driver genes, BRAF, NRAS, and NF1 in the MAPK pathway, were lower but exhibited a stronger positive correlation as compared with low-risk patients. Moreover, the genes involved in nicotinamide adenine dinucleotide metabolism were negatively correlated with the expression of BRAF in the high-risk group. Function analysis revealed that the upregulated genes in the high-risk group were enriched in the cytochrome P450-mediated metabolism of chemical carcinogens. Furthermore, the low-risk group had high levels of gamma delta T cells infiltration, while regulatory T cells were accumulated in the high-risk group. The present study offers a promising new prognostic signature for CM, and provides insight into the mechanisms of melanoma progression.


Measurement and analysis of cohort-size variations.
"This article discusses two measures of cohort-size variations: the age-standardized cohort size, and the growth rate of the number of births.... The usefulness of the two measures in analysing recent demographic trends is demonstrated....[The author] discusses meanings and usefulness of the ratio of the crude birth rate (CBR)/total fertility rate (TFR) in studying the effects of age structure changes on population growth." The geographical scope is worldwide, with particular attention to China, Indonesia, Japan, the Philippines, and Thailand.


Medical Therapy for Eisenmenger Syndrome: A Case Report and Review of Literature.
Eisenmenger syndrome (ES) is defined as the process in which a long-standing left-to-right cardiac shunt caused by a congenital heart defect is eventually reversed into a cyanotic right-to-left shunt. It is crucial to recognize this grave pathology at the earliest because once it develops, treatment by medical or surgical means becomes even more challenging. In past decades, various therapeutic options have been developed that address the specific pathophysiological aspects of the disease and have shown to improve functional capacity and quality of life. There are three major therapeutic pathways in pulmonary hypertension treatment - endothelin receptor antagonists, phosphodiesterase type-5 inhibitors, and prostacyclin derivatives. These therapies not only improve hemodynamic parameters and exercise capacity but they also improve prognosis with various form of Pulmonary hypertension including ES. We report a case of a 35-year-old female of ostium secundum atrial septal defect with ES, started on upfront combination therapy of ambrisentan and tadalafil who demonstrated marked improvement after 3 months of medical therapy.


[Propafenone and flecainide in the therapy of ventricular arrhythmias].
Flecainide and propafenone are antiarrhythmic drugs of the class 1C (Vaughan and Williams) commonly used for ventricular arrhythmias. The purpose of the present study was to evaluate the efficacy of these drugs in 170 consecutive patients with ventricular arrhythmias who referred to our cardiological ambulatory. The study population was divided into two groups according to the absence (group A,82 patients) or presence of organic heart disease (group 1B: 51 patients with left ventricular ejection fraction (LVEF) >35%; group 2B: 37 patients with LVEF<35%). Ventricular arrhythmias were evaluated with a 48 hours Holter monitoring at baseline, and with a control 24 hours Holter monitoring at 15 days (for optimizing the dosage), at 5 months and at 10 months from the beginning of antiarrhythmic therapy. Patients of group A were randomly assigned to antiarrhythmic treatment (flecainide 150-300 mg/die or propafenone 450-900 mg/die). For patients of group B, such choice was leaded by the clinical and strumental data (32 patients were treated with flecainide, 56 patients with propafenone). In the 160 patients who ended the 10 months follow-up, we observed the following results: patients of group A showed a mean percentage reduction in incidence of premature ventricular complexes (PVC) after therapy in comparison to basal conditions of 93% and 89% with flecainide and propafenone, respectively, after a treatment of 5 months (p < 0.001); after 10 months mean percentage reduction of PVC was 91% with each drug (p = n.s.); complex ventricular events (CVE) were reduced of 90% and of 100% after 5 and 10 months, respectively, of treatment with flecainide and of 100% both after 5 and 10 months of treatment with propafenone (p = n.s.) -- patients of group 1B showed a mean percentage reduction of PVC of 87% and 84% after 5 and 10 months, respectiively, of treatment with propafenone (p = n.s.); after 5 months of therapy mean percentage CVE reduction was 66% with flecainide and 86% with propafenone (p < 0.001); after 10 months this mean reduction was 53% with flecainide and 73% with propafenone (p < 0.001). -- patients of group 2B showed a mean reduction of PVC of 59% and 58% after 5 and 10 months of therapy with flecainide, and of 65% and 67% after 5 and 10 months of therapy with propafenone (p = n.s.); CVE were reduced of 28% with flecainide and of 47% with propafenone after 5 months of treatment (p < 0.001) and of 36% with flecainide against 52% with propafenone after 10 months (p < 0.01). In the present study there was no significant difference between the two drugs in terms of tollerance and collateral effects (8% with flecainide vs 7%, with propafenone). Our results confirm the efficacy of the 1C class drugs in the treatment of "essential" ventricular arrhytmias. This efficacy appears reduced in non selected patients with organic heart disease. In these latter patients propafenone has shown more efficacy than flecainide in reducing CVE.


Effectiveness of bronchial thermoplasty in patients with severe refractory asthma: Clinical and histopathologic correlations.
The effectiveness of bronchial thermoplasty (BT) has been reported in patients with severe asthma, yet its effect on different bronchial structures remains unknown. We sought to examine the effect of BT on bronchial structures and to explore the association with clinical outcome in patients with severe refractory asthma. Bronchial biopsy specimens (n = 300) were collected from 15 patients with severe uncontrolled asthma before and 3 months after BT. Immunostained sections were assessed for airway smooth muscle (ASM) area, subepithelial basement membrane thickness, nerve fibers, and epithelial neuroendocrine cells. Histopathologic findings were correlated with clinical parameters. BT significantly improved asthma control and quality of life at both 3 and 12 months and decreased the numbers of severe exacerbations and the dose of oral corticosteroids. At 3 months, this clinical benefit was accompanied by a reduction in ASM area (median values before and after BT, respectively: 19.7% [25th-75th interquartile range (IQR), 15.9% to 22.4%] and 5.3% [25th-75th IQR], 3.5% to 10.1%, P < .001), subepithelial basement membrane thickening (4.4 μm [25th-75th IQR, 4.0-4.7 μm] and 3.9 μm [25th-75th IQR, 3.7-4.6 μm], P = 0.02), submucosal nerves (1.0 ‰ [25th-75th IQR, 0.7-1.3 ‰] immunoreactivity and 0.3 ‰ [25th-75th IQR, 0.1-0.5 ‰] immunoreactivity, P < .001), ASM-associated nerves (452.6 [25th-75th IQR, 196.0-811.2] immunoreactive pixels per mm2 and 62.7 [25th-75th IQR, 0.0-230.3] immunoreactive pixels per mm2, P = .02), and epithelial neuroendocrine cells (4.9/mm2 [25th-75th IQR, 0-16.4/mm2] and 0.0/mm2 [25th-75th IQR, 0-0/mm2], P = .02). Histopathologic parameters were associated based on Asthma Control Test scores, numbers of exacerbations, and visits to the emergency department (all P ≤ .02) 3 and 12 months after BT. BT is a treatment option in patients with severe therapy-refractory asthma that downregulates selectively structural abnormalities involved in airway narrowing and bronchial reactivity, particularly ASM, neuroendocrine epithelial cells, and bronchial nerve endings.


Terlipressin-induced Peripheral Ischemic Gangrene in a Diabetic Patient.
Terlipressin is used commonly in the management of hepatorenal syndrome and acute variceal bleeding. Like its parent compound vasopressin, it is also notorious for its ischemic complications. Terlipressin-induced ischemic complications can virtually affect any part of the body, but the incidence of serious complications is less than its parent compound vasopressin. Here, we report a case of terlipressin-induced peripheral ischemic gangrene in a diabetic male, which ultimately led to death of the patient.


SPINK5 is a Prognostic Biomarker Associated With the Progression and Prognosis of Laryngeal Squamous Cell Carcinoma Identified by Weighted Gene Co-Expression Network Analysis.
Laryngeal squamous cell carcinoma (LSCC) is one of the most common types of head and neck squamous cell carcinomas (HNSCC) and is the second most prevalent malignancy occurring in the head and neck or respiratory tract, with a high incidence and mortality rate. Survival is limited for patients with LSCC. To identify more biomarkers associated with the prognosis of patients with LSCC, using bioinformatics analysis, this study used The Cancer Genome Atlas (TCGA) LSCC dataset and gene expression profiles of GSE59102 from the Gene Expression Omnibus (GEO). Eighty-one differentially co-expressed genes were identified by weighted gene co-expression network analysis (WGCNA). Next, 10 hub genes (PPL, KRT78, CRNN, PTK7, SCEL, AGRN, SPINK5, AIF1L, EMP1, and PPP1R3C) were screened from a protein-protein interaction (PPI) network. Based on survival analysis, SPINK5 was significantly correlated with survival time in LSCC patients. After verification in the TCGA and HPA databases, SPINK5 was selected as a prognostic biomarker. Finally, the GSEA results showed that downregulation of SPINK5 gene expression may promote tumorigenesis and the development of cancers by the "BASAL CELL CARCINOMA" pathway, and it has been implicated in disrupting DNA damage and repair pathways. Collectively, SPINK5 may serve as a potential prognostic biomarker in LSCC.


Leaching efficiency of copper from industrial sludge with traditional acid extraction (TAE) and microwave assisted treatment (MAT).
In this study various leaching technologies were applied to extract heavy metals from sludge from the printed circuit board process. These methods included traditional acid extraction (TAE) and microwave assisted treatment (MAT). The target heavy metal was copper. Experiments were performed to determine the acid the leaching efficiency by changing operating conditions, including acid type and concentration, leaching time, microwave power, and solid to liquid ratio (S/L). The TAE method demonstrated that copper leaching from a sludge of fine particle sizes (d < 150 microm) was 74% efficient at an S/L of 1/6, a leaching time of 18 hr, using nitric acid in the leaching solution. Then, when the microwave powers were 800 W and 400 W; the leaching time was 30 mins, and the coarse particle size (d < 9.5 mm) was by 1 N sulfuric acid, the copper leaching effects were 93% and 77%, respectively. The MAT procedure reduced the leaching time and improved the leaching efficiency above that obtained by TAE.


Does glyceryl nitrate prevent post-ERCP pancreatitis? A prospective, randomized, double-blind, placebo-controlled multicenter trial.
Acute pancreatitis is the most dreaded complication of ERCP. Two studies have shown a significant effect of glyceryl nitrate (GN) in preventing post-ERCP pancreatitis (PEP). We wanted to evaluate this promising effect in a larger study with a realistically precalculated incidence of PEP. A randomized, double-blind, placebo-controlled multicenter study including patients from 14 European centers was performed. A total of 820 patients were entered; 806 were randomized. The active drug was transdermal GN (Discotrine/Minitran, 3M Pharma) 15 mg/24 hours; placebo (PL) was an identical-looking patch applied before ERCP. A total of 401 patients received GN; 405 received PL. Forty-seven patients had PEP (5.8%), 18 (4.5%) in the GN group and 29 (7.1%) in the PL group. The relative risk reduction of PEP in the GN group of 36% (95% CI, 11%-65%) compared with the PL group was not statistically significant (P = .11). Thirteen had mild pancreatitis (4 in the GN group, 9 in the PL group), 26 had moderate pancreatitis (9 in the GN group, 17 in the PL group), and 8 had severe pancreatitis (5 in the GN group, 3 in the PL group). Headache (P < .001) and hypotension (P = .006) were more common in the GN group. Significant variables predictive of PEP were not having biliary stones extracted; hypotension after ERCP; morphine, propofol, glucagon, and general anesthesia during the procedure; or no sufentanil during the procedure. The trial showed no statistically significant preventive effect of GN on PEP. Because of a considerable risk of a type II error, an effect of GN may have been overlooked. (ClinicalTrials.gov ID: NCT00121901.).


GNIFdb: a neoantigen intrinsic feature database for glioma.
Neoantigens are mutation-containing immunogenic peptides from tumor cells. Neoantigen intrinsic features are neoantigens' sequence-associated features characterized by different amino acid descriptors and physical-chemical properties, which have a crucial function in prioritization of neoantigens with immunogenic potentials and predicting patients with better survival. Different intrinsic features might have functions to varying degrees in evaluating neoantigens' potentials of immunogenicity. Identification and comparison of intrinsic features among neoantigens are particularly important for developing neoantigen-based personalized immunotherapy. However, there is still no public repository to host the intrinsic features of neoantigens. Therefore, we developed GNIFdb, a glioma neoantigen intrinsic feature database specifically designed for hosting, exploring and visualizing neoantigen and intrinsic features. The database provides a comprehensive repository of computationally predicted Human leukocyte antigen class I (HLA-I) restricted neoantigens and their intrinsic features; a systematic annotation of neoantigens including sequence, neoantigen-associated mutation, gene expression, glioma prognosis, HLA-I subtype and binding affinity between neoantigens and HLA-I; and a genome browser to visualize them in an interactive manner. It represents a valuable resource for the neoantigen research community and is publicly available at http://www.oncoimmunobank.cn/index.php. http://www.oncoimmunobank.cn/index.php.


Impact of changes in psychological resilience during treatment with intensity-modulated radiotherapy on nasopharyngeal carcinoma patients: a prospective study.
Nasopharyngeal carcinoma (NPC) patients can undergo changes in psychological status during treatment. The aim of this prospective study was to determine the impact of the changes in psychological resilience on the quality of life (QoL) and long-term outcomes of patients. Patients with NPC receiving intensity-modulated radiotherapy (IMRT) between March 2017 and February 2019 were prospectively included. Their psychological resilience was evaluated by the Connor-Davidson resilience scale (CD-RISC) twice. Patients were then divided into the improved psychological resilience group and the deteriorated group. All patients were followed up for at least 2 years, and acute or late severe complications were recorded. The QoL of patients was evaluated within 1 year by the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30-questions (QLQ-C30) and the Head and Neck 35-questions (HN35). Logistic regression analysis was used for the analysis of risk factors of psychological resilience in NPC patients. Similarly, linear regression analysis was used for the analysis of risk factors of QoL in NPC patients. The overall survival rate and progression-free survival rate were recorded and compared between the 2 groups using Kaplan-Meier curves. A total of 180 patients were included. The mean CD-RISC scores before radiotherapy and after radiotherapy were 55.8±7.0 and 58.4±7.8 points, respectively. Patients were divided into 104 patients in the improved group and 76 patients in the deteriorated group. Older age, advanced stage, chemotherapy treatment, and severe complications were important risk factors according to the multivariable logistic regression analysis. There were no significant differences in QLQ-C30 and HN35 scores before radiotherapy between the 2 groups, while significant differences were found in most items in the QLQ-C30 and HN35 between the 2 groups. Deteriorated resilience was identified as an important risk factor of QoL according to the multivariable linear regression analysis. NPC patients had significantly higher overall survival and progression-free survival in the improved group than in the deteriorated group. Psychological resilience has an important impact on the prognosis of NPC patients, thus more attention should be paid to their psychological status during treatment with radiotherapy.


Decreased expression of multidrug efflux transporters in the brains of GSK-3beta transgenic mice.
Multidrug efflux transporters protect cells in the brain from potentially harmful substances but also from therapeutically useful drugs. Thus any condition that causes changes in their expression is of some importance with regard to drug access. In this study, changes in efflux transporter expression are investigated in mice containing a mutant constitutively active glycogen synthase kinase-3 (GSK-3beta) transgene, driven by the Thy-1 promoter so limiting its localization predominantly to neurons and some glial cells. As expected, decreases in beta-catenin were evident via Western blot analyses of cortical homogenates prepared from brains of these transgenic mice. As assessed by real time qRT-PCR, decreased transcript levels of the mdr1b isoform of P-glycoprotein, Mrp1 and Mrp4, (transporters associated with neurons and/or glial cells) were observed in the cortex but not the subventricular zone or hippocampus of the transgenic compared to wild type mouse brains. By contrast, no such decreases were evident with the mdr1a isoform of P-glycoprotein and Bcrp, transporters predominantly found in brain endothelium. Such transporter expression changes could not be accounted for by alterations in blood vessel density or neuronal to glial cell ratios as analyzed both from immunocytochemical staining and from RT-PCR. These observations support previous in vitro data showing that manipulations to GSK-3beta activity that alter signaling via beta-catenin can influence the expression of efflux transporters. Implications from this are that drug distribution into cells within the brain of these transgenic mice could be enhanced, hence warranting further investigation.


68Ga-PSMA Uptake in Hepatocellular Carcinoma.
Ga-PSMA based integrated PET/MRI is emerging as a novel imaging technique for the staging of prostate carcinoma. We report a case of a 77-year-old man with raised prostate-specific antigen who presented to us for Glu-NH-CO-NH-Lys-(Ahx)-[Ga-(HBED-CC)] (Ga-PSMA) simultaneous PET/MRI scan for prostate cancer evaluation. A PSMA avid hepatic lesion on the background of cirrhotic liver was noted apart from PSMA avid lesion in the peripheral gland of the prostate. On histopathological examination, the hepatic lesion turned out to be hepatocellular carcinoma.


A comparison of individual genotyping and pooled DNA analysis for polymorphism validation prior to large-scale genetic studies.
Polymorphism validation is an important issue in genetic studies because only polymorphic markers provide useful information. We analyzed genetic data for 180 SNPs in the human major histocompatibility complex region in Caucasian and Taiwanese populations, and evaluated ethnic heterogeneity between these populations to illustrate the importance of polymorphism validation. An initial individual genotyping experiment (IGE) with 95 samples was compared with a DNA pooling allele-typing experiment (PAE) of 630 individuals for polymorphism validation based on authentic data sets. Afterwards, all samples were genotyped individually in a confirmation study. Under narrow (broad) polymorphism criteria, 24 (41) polymorphic SNPs in Caucasians could not be validated in the Taiwanese population, suggesting a 13% (23%) inconsistency rate and revealing a strong discrepancy between genetic backgrounds, probably due to ethnic heterogeneity. IGE yielded high sensitivity and specificity for polymorphism validation, but may be sensitive to sampling variation. PAE showed high sensitivity (97%) and specificity (100%) using a narrow polymorphism criterion, but reduced specificity (83%) using a broad criterion. Public domain polymorphism databases should therefore be used with caution and polymorphism validation should be performed routinely prior to conducting large-scale genetic studies. PAE is a cost-saving, reliable alternative to IGE for polymorphism validation, especially for a stringent polymorphism criterion.


Memory performance in multiple sclerosis patients correlates with central brain atrophy.
To assess whole brain and central brain atrophy as well as their differential relation to memory, cognitive performance, fatigue, depression and quality of life in patients with relapsing-remitting multiple sclerosis (RRMS). A 3D flow compensated gradient recalled T1-weighted MRI was acquired in 45 RRMS patients. An automated analysis tool was used to calculate brain parenchymal fraction (BPF) and ventricular brain fraction (VF). All patients were assessed with neuropsychological tests focusing on memory and self-rating scales for depression, fatigue and quality of life. Age corrected partial correlations between brain atrophy, motor performance, psychological scales and test scores were calculated. BPF correlated moderately (0.3 < or = r < 0.5) with duration of symptoms and disease, the Expanded Disability Status Scale (EDSS), the upper extremity motor performance, and with mental aspects of quality of life. VF correlated moderately with EDSS, upper and lower extremity motor performance and memory functions. Neither BPF nor VF correlated with fatigue and depression. Results of several cognitive tests correlated moderately with depression and fatigue, the Paced Auditory Serial Addition Test (PASAT) showing the largest correlation. Memory performance shows a correlation with relative ventricular size in RRMS patients, indicating the strategic location of the ventricle system along the structures of the limbic system and its vulnerability in MS. The PASAT and several other cognitive tests show moderate correlations with depression and fatigue, arguing for an inter relation between the cognitive functioning and the emotional state of patients. However, this relation is independent of measurable brain atrophy.


Evolution properties of the intensity distribution of an anisotropic Gaussian Schell-model beam in a turbulent atmosphere.
An analytical expression for the cross-spectral density of an anisotropic Gaussian Schell-model (AGSM) beam propagating in a turbulent atmosphere is derived, which is featured by its clear physical meaning. The evolution properties of the intensity distribution of an AGSM beam in a turbulent atmosphere are studied thoroughly. It is found that owing to the anisotropy of the source coherence, the intensity distribution can have many different evolution processes; but under the influence of the turbulent atmosphere, it will finally take on a circular shape. The effects of lowering the source coherence on the circularization speed of the intensity distribution are also investigated. It is found that, when the anisotropy of the source coherence is taken into account, lowering the source coherence can accelerate or decelerate the circularization of the intensity distribution. We propose and demonstrate five kinds of conditions, and, under each condition, lowering the source coherence has a unique and definite effect on the circularization speed of the intensity distribution. In particular, we give an analysis about the equivalence between the problem studied in one of our five conditions and that in the work of Cai and He [Appl. Phys. Lett. 89, 041117 (2006)] and show that their relevant conclusion is incorrect.


Cytokine elevation and transaminitis after laparoscopic donor nephrectomy.
Acute kidney injury frequently occurs in the critically ill and often progresses into multiorgan dysfunction syndrome, resulting in high mortality. We previously showed that nephrectomized mice had increased interleukin (IL)-6 and tumor necrosis factor (TNF)-α that directly contributed to systemic inflammation and hepatic injury. In this study, we examined whether patients undergoing laparoscopic donor nephrectomy have increased postoperative cytokine levels with injury to the liver and whether the remaining kidney sustains injury. Serial serum and urine samples were collected from 32 patients undergoing laparoscopic donor nephrectomy and 17 patients undergoing nonrenal laparoscopic surgery. Serum IL-6, IL-18, TNF-α and monocyte chemotactic protein-1 (MCP-1) (markers of systemic inflammation) and urinary neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), MCP-1, and IL-18 (markers of acute kidney injury) were quantified by enzyme-linked immunosorbent assay. We also analyzed serum creatinine, aspartate transaminase (AST), and alanine transaminase to assess liver injury. Patients who underwent donor nephrectomy not only demonstrated increased serum creatinine but also had significant increases in serum IL-6, MCP-1, and AST. Serum TNF-α also trended upward in donor nephrectomy patients. Finally, the donor nephrectomy group showed increased urinary NGAL but not KIM-1 at 24 h. Taken together, our findings of increased serum IL-6, MCP-1, and AST after donor nephrectomy suggest that an acute reduction of kidney function induces systemic inflammation and may have distant effects on the liver. Further studies are needed to correlate increased urinary NGAL after donor nephrectomy both as a potential marker for renal tubular stress and/or hypertrophy in the contralateral kidney.


Oxygen uptake and plasma catecholamines during submaximal and maximal exercise after long-term beta-receptor blockade.
Beta-receptor antagonists lower exercise heart rate and cardiac output, and can thus be expected to interfere with oxygen transport, and hence physical performance, particularly at higher levels of activity. Therefore, the effects of a 4-week and 15-month treatment period with the beta 1-selective receptor blocker acebutolol (500 mg daily) on oxygen uptake and plasma catecholamines during submaximal steady-state and maximal exercise and on maximal work load were studied in eight WHO stage 1 hypertensive men (mean age 36.4 years). Oxygen uptake, ventilation, and plasma noradrenaline, adrenaline, and dopamine concentrations during steady-state exercise were not significantly different from control conditions either after 4 weeks or after 15 months of receptor blockade, although heart rates were significantly (27% and 25%, respectively; P less than 0.01) reduced. After the 4-week treatment period, maximal oxygen uptake (3.9% reduction, NS) and maximal work load (2.4% reduction, NS) tended to be slightly lower after acebutolol compared with control values; maximal oxygen pulse was significantly (P less than 0.05) increased. However, after long-term treatment of 15 months, maximal oxygen uptake was virtually identical compared with pretreatment values, and maximal work load tended to be higher (5.2%, NS); plasma noradrenaline and adrenaline concentrations were significantly (P less than 0.05) enhanced. Since beta 1-selective receptor blockers do not affect maximal oxygen uptake and maximal work capacity after long-term treatment, they appear preferable for patients taking part in preventive and rehabilitative training programs.


Postnatal maturation of the Kallikrein-kinin system in the rat kidney: from enzyme activity to receptor gene expression.
During the course of aging, the balance between intrarenal hormones is disturbed. These age-related changes are well documented for the vasoconstrictor renin-angiotensin system, but comparable information on the renal kallikrein-kinin system is not yet available. The status of the kallikrein-kinin system was assessed by (1) kallikrein activity, measured by RIA; (2) maximum binding site density (Bmax) and affinity (Kd) of nonapeptide bradykinin (BK)-2 receptor, estimated by binding assays; (3) expression of BK2-receptor receptor mRNA, detected by reverse transcription-polymerase chain reaction (RT-PCR) using specific BK2-oligonucleotide primers. These parameters were determinated on renal glomeruli of 3-, 5-, 8-, 12- and 38-wk-old normotensive rats. Kallikrein activity increased from 3.2 to 7.7 ng BK/min per mg protein. The density of BK2 binding sites also rose from 12 to 40 femtomoles/mg protein with no difference in affinity. There was no change in specificity, which remained that expected of a BK2 receptor. The increase in the density of BK2 binding sites was associated with an augmented mRNA expression, whereas beta-actin mRNA used as a control remained unchanged. The ratio of BK2 mRNA to beta-actin mRNA indicated maximum steady expression after 8 wk of age. The data provide evidence that the renal kallikrein-kinin system develops postnatally.


Utility of KRAS mutation detection using circulating cell-free DNA from patients with colorectal cancer.
In this study, we evaluated the clinical utility of detecting KRAS mutations in circulating cell-free (ccf)DNA of metastatic colorectal cancer patients. We prospectively recruited 94 metastatic colorectal cancer patients. Circulating cell-free DNA was extracted from plasma samples and analyzed for the presence of seven KRAS point mutations. Using the Invader Plus assay with peptide nucleic acid clamping method and digital PCR, KRAS mutations were detected in the ccfDNA in 35 of 39 patients previously determined to have primary tumors containing KRAS mutations using the Luminex method, and in 5 of 55 patients with tumors containing wild-type KRAS. Curative resection was undertaken in 7 of 34 patients with primary and ccfDNA KRAS mutations, resulting in the disappearance of the mutation from the cell-free DNA in five of seven patients. Three of these patients had tumor recurrence and KRAS mutations in their ccfDNA reappeared. Epidermal growth factor receptor blockade was administered to 24 of the KRAS tumor wild-type patients. Of the 24 patients with wild-type KRAS in their primary tumors, three patients had KRAS mutations in their ccfDNA and did not respond to treatment with epidermal growth factor receptor (EGFR) blockade. We also detected a new KRAS mutation in five patients during chemotherapy with EGFR blockade, before disease progression was detectable with imaging. The detection of KRAS mutations in ccfDNA is an attractive approach for predicting both treatment response and acquired resistance to EGFR blockade, and for detecting disease recurrence.


Oral effervescent agent improving magnetic resonance cholangiopancreatography.
The purpose of our study was to validate the oral effervescent agent improving magnetic resonance cholangiopancreatography (MRCP) in patients with suspicious pancreatobiliary disease. One hundred and eleven consecutive patients with alleged or suspected pancreatobiliary tree problems who had undergone two-dimensional (2D) MRCP imaging both before and after oral effervescent enhancement (conventional-MRCP and enhanced-MRCP) were included. Two radiologists independently scored overall image quality, visualization of ten ductal segments, and gastroduodenal fluid signal intensity score. In consensus, they assessed the presence of gastroduodenal fluids and pancreatobiliary tree overlapping. The data were analyzed using Wilcoxon's signed-rank test, McNemar test, and paired t-test. The grades of overall image quality and individual biliary duct visualization for ten targeted ductal segments, and gastroduodenal fluid signal intensity scores increased significantly on enhanced-MRCP by both readers (P≤0.02), but there was no significant increase for pancreatic duct (PD) at head and tail. On enhanced-MRCP, gastroduodenal fluids except for gastric fundus were less detected rather than those on conventional-MRCP. Anatomic structures of gastroduodenal fluids overlapping extrahepatic bile duct were mainly gastric antrum, duodenal bulb, and 2nd portion on conventional-MRCP. However, these fluids were less overlapped on enhanced-MRCP (P<0.001). Gastric body and antrum were main anatomic structures of gastroduodenal fluids overlapping PD on conventional-MRCP, and fluid in these locations significantly less overlapped PD on enhanced-MRCP (P≤0.02). Oral administration of effervescent agent provided effective elimination of gastroduodenal fluid overlapping pancreatobiliary ductal system at MRCP and can improve the quality of the examination in the patients with known or suspected pancreatobiliary disease.


Elevated sialyl Lewis X-i antigen levels in pleural effusions in patients with carcinomatous pleuritis.
The levels of sialyl Lewis X-i antigen (SLX), which is one of the cancer-associated carbohydrate antigens, were evaluated in 83 malignant and 46 benign pleural effusions. SLX levels in pleural effusion due to lung adenocarcinoma were significantly higher than those due to benign diseases (p < 0.0001), lung cancer other than adenocarcinoma (p = 0.0052), and adenocarcinoma originating from other organs (p = 0.0492). According to receiver operating characteristic (ROC) curve analysis, the optimal cut-off level in the discrimination between malignant and benign pleural effusions was 92 U/ml, which gave a sensitivity of 57.1% and a specificity of 77.8%. The cut-off level of pleural effusion in patients with carcinomatous pleuritis might be higher than that of serum (38 U/ml).


Characterization of the N-acetyl-α-D-glucosaminyl l-malate synthase and deacetylase functions for bacillithiol biosynthesis in Bacillus anthracis .
Bacillithiol (Cys-GlcN-malate, BSH) has recently been identified as a novel low-molecular weight thiol in Bacillus anthracis, Staphylococcus aureus, and several other Gram-positive bacteria lacking glutathione and mycothiol. We have now characterized the first two enzymes for the BSH biosynthetic pathway in B. anthracis, which combine to produce α-d-glucosaminyl l-malate (GlcN-malate) from UDP-GlcNAc and l-malate. The structure of the GlcNAc-malate intermediate has been determined, as have the kinetic parameters for the BaBshA glycosyltransferase (→GlcNAc-malate) and the BaBshB deacetylase (→GlcN-malate). BSH is one of only two natural products reported to contain a malyl glycoside, and the crystal structure of the BaBshA-UDP-malate ternary complex, determined in this work at 3.3 Å resolution, identifies several active-site interactions important for the specific recognition of l-malate, but not other α-hydroxy acids, as the acceptor substrate. In sharp contrast to the structures reported for the GlcNAc-1-d-myo-inositol-3-phosphate synthase (MshA) apo and ternary complex forms, there is no major conformational change observed in the structures of the corresponding BaBshA forms. A mutant strain of B. anthracis deficient in the BshA glycosyltransferase fails to produce BSH, as predicted. This B. anthracis bshA locus (BA1558) has been identified in a transposon-site hybridization study as required for growth, sporulation, or germination [Day, W. A., Jr., Rasmussen, S. L., Carpenter, B. M., Peterson, S. N., and Friedlander, A. M. (2007) J. Bacteriol. 189, 3296-3301], suggesting that the biosynthesis of BSH could represent a target for the development of novel antimicrobials with broad-spectrum activity against Gram-positive pathogens like B. anthracis. The metabolites that function in thiol redox buffering and homeostasis in Bacillus are not well understood, and we present a composite picture based on this and other recent work.


Chrysosporium anamorph of Nannizziopsis vriesii associated with fatal cutaneous mycoses in the salt-water crocodile (Crocodylus porosus).
The Chrysosporium anamorph of Nannizziopsis vriesii, recently identified as the cause of cutaneous infections in chameleons and brown tree snakes, was associated with skin infections and deaths in salt-water crocodile (Crocodylus porosus) hatchlings on two separate occasions 3 years apart. In all, 48 animals died from the infection. All hatchlings came from the same farm in northern Queensland, Australia.


Fabrication and characterization of a pH-sensitive intelligent film incorporating dragon fruit skin extract.
A novel intelligent pH-sensing indicator based on gelatin film and anthocyanin extracted from dragon fruit skin (Hylocereus polyrhizus) (DFSE) as a natural dye was developed to monitor food freshness by the casting method. Anthocyanin content of DFSE was 15.66 ± 1.59 mg/L. Dragon fruit bovine gelatin films were characterized by Fourier transform infrared spectroscopy (FTIR) and observed by a scanning electron microscope (SEM). Moisture content, mechanical properties, water solubility, water vapor permeability (WVP), light transmittance, color, and pH-sensing evaluations were evaluated for potential application. FTIR spectroscopy revealed that the extracted anthocyanin could interact with the other film components through hydrogen bonds. When the extract was added, films showed a smooth and clear surface as observed by SEM. The addition of anthocyanin increased the moisture content, thickness, and water solubility of the films, but decreased the WVP and light transmittance of films. Also, the incorporation of 15% v/v DFSE decreased the tensile strength from 17.04 to 12.91 MPa, increasing the elongation at break from 91.19% to 107.86%. The films showed higher ΔE with increasing DFSE content, which indicated that the film had good color variability. A significant difference in the color of the films was observed with exposure to different pH buffer solutions. The findings demonstrated that gelatin film incorporated with DFSE could be used as a visual indicator of pH variations to monitor the freshness of foods during storage time.


Outbreak of louse-borne relapsing fever in Jimma, south western Ethiopia.
Some epidemiological aspects of louse-borne relapsing fever during the epidemic of 1991 in Ethiopia are described. There was male preponderance and the age ranged from two months to 58 years (mean, sd = 18.05,11.19). There was no sex differential in children. Students, soldiers and jobless contributed the majority. There were two peaks (August and March) in the seasonal pattern. About two-thirds was infested with lice and significant associations were found among adults, soldiers, ethnicity, illiterate persons and family size above seven. The household attack rate was 15.1%, the attack being higher among children. Promotion of preventive measures are recommended.


[Childbirth as I see it . . . or the way I wish it was? Expectations of pregnant women towards childbirth and obstetric care in the public health care system].
Explanations for increased cesarean section rates in Brazil have focused on the organization of obstetric care, training of health professionals, and women's demand for surgical deliveries. This study aimed to identify pregnant women's expectations towards childbirth. Three focus groups were conducted in a public hospital in the city of São Paulo. Analytical categories were: vaginal birth, forceps, c-section, prenatal care, and obstetric care. The desire for c-sections was associated with a demand for tubal ligation, and although women feared labor pains, they were more afraid of how the obstetric team might react to their complaints. Lack of information on reproductive issues was associated with a demand for more information. There was a preference for vaginal births, since most women feared c-sections due to risks associated with this surgical intervention. The authors propose that the demand for cesareans among women should be reconsidered as one of the main factors in the rise in surgical deliveries in the Brazilian health care system.


Occupational mortality from squamous cell carcinoma of the esophagus in the United States during 1991-1996.
The epidemiology of esophageal squamous cell cancer has remained poorly understood. The occupational distribution of this cancer may provide clues about its yet unknown etiology. Data files from the National Center for Health Statistics (NCHS) of the United States offer a unique source to study causes of death, broken down by occupation and industry. The number of deaths from esophageal cancer was retrieved from the computerized US vital statistics. Mortality by occupation or industry was expressed as standardized proportional mortality ratio (PMR), adjusted by age, gender, and ethnicity. Between 1991 and 1996, 63,717 subjects died from esophageal squamous cell carcinoma. Mortality was particularly high among nonwhites and men. The industrial and the occupational distributions shared a similar pattern. Mortality from esophageal squamous cell carcinoma occurred more frequently among subjects exposed to silica dust, such as brickmasons and stonemasons, concrete and terrazzo finishers, roofers, and construction laborers. It was also high in such industries as unspecified machinery or manufacturing and such occupations as unspecified material handlers, janitors, or cleaners. It was low in industries and occupations associated with agriculture, clergy, work in religious organizations, and textiles. In conclusion, mortality from esophageal squamous cell carcinoma appeared to be low in occupations associated with less consumption of alcohol and tobacco. It was high among occupations potentially associated with exposure to silica dust and chemical solvents or detergents.


Real-time nondestructive evaluation of fiber composite laminates using low-frequency Lamb waves.
Amid the nondestructive evaluation techniques available for the inspection of composite materials, only a few are suitable for implementation while the component is in service. The investigation examines the application of Lamb waves at low-frequency-thickness products for the detection of delaminations in thick composite laminates. Surface-mounted piezoelectric devices were excited with a tone burst to generate elastic waves in the structure. Experiments were carried out on composite beam specimens where wave propagation distances over 2 m were achieved and artificially induced delaminations as small as 1 cm2 were successfully identified. The feasibility of employing piezoelectric devices for the development of smart structures, where a small and lightweight transducer system design is required, has been demonstrated. The resonance spectrum method, which is based on the study of spectra obtained by forced mechanical resonance of samples using sine-sweep excitation, has been proposed as a technique for measuring the A0 Lamb mode phase velocity. The finite-element method was also used to investigate qualitatively the dynamic response of laminates to wave propagation. Several locations and spatial distribution of the actuators were examined showing the advantages of using transducers arrays for the inspection of large structures.


[Chronic necrotizing pulmonary aspergillosis: diagnosis and treatment].
The purpose of this study was to describe the approaches to the diagnosis and the treatment of chronic necrotizing pulmonary aspergillosis (CNPA). The study included 8 patients with CNPA confirmed by histopathological studies. The patients were admitted to the First Affiliated Hospital of College of Medicine of Zhejiang University during 2000-2004. The clinical data were analyzed, and related literature was reviewed. The clinical, radiological and pathological characteristics, and the approaches to the diagnosis and the treatment were described. CNPA was a rare form of pulmonary aspergillosis. Common presentations included productive cough (8/8), haemoptysis (7/8) and constitutional symptoms (5/8). Radiographically, the air crescent sign was suggestive of the diagnosis, which was found in 4 patients. Consolidation was also common in the other 4 cases. The histological feature of CNPA included the presence of tissue invasion by Aspergillus, tissue necrosis and granulomatous inflammation. The clinical manifestations of CNPA are non-specific. Radiography plays a pivotal role in the diagnosis of CNPA. Treatment with antifungal medications is indicated once the diagnosis is made. Pulmonary resection should be considered when patients have prolonged illness or frequent haemoptysis.


Smad3 binds Scleraxis and Mohawk and regulates tendon matrix organization.
TGFβ plays a critical role in tendon formation and healing. While its downstream effector Smad3 has been implicated in the healing process, little is known about the role of Smad3 in normal tendon development or tenocyte gene expression. Using mice deficient in Smad3 (Smad3(-/-) ), we show that Smad3 ablation disrupts tendon architecture and has a dramatic impact on normal gene and protein expression during development as well as in mature tendon. In developing and adult tendon, loss of Smad3 results in reduced protein expression of the matrix components Collagen 1 and Tenascin-C. Additionally, when compared to wild type, tendon from adult Smad3(-/-) mice shows a down regulation of key tendon marker genes. Finally, we have established that Smad3 has the ability to physically interact with the critical transcriptional regulators Scleraxis and Mohawk. Together these results indicate a central role for Smad3 in normal tendon formation and in the maintenance of mature tendon.


Polysomes, Messenger RNA, and Growth in Soybean Stems during Development and Water Deficit.
The polysome status and populations of polysomal mRNA were examined in different regions of dark-grown soybean (Glycine max [L.] Merr.) stems that contained either dividing, elongating, or mature (nongrowing) cells. There was a developmental gradient of polysome content in which the dividing tissue had the highest levels and the mature tissue the lowest. A few hours after transplanting the seedlings to vermiculite having low water content (water potential Psi(w) = -0.29 megapascals), stem growth rate decreased to 30% of well-watered controls and the polysome content decreased most in the dividing and elongating tissues. After 24 to 36 hours, stem growth and polysome content recovered gradually. In vitro translation products of polysomal mRNA from dividing, elongating or mature tissue were examined on two-dimensional gels. In well-watered controls, each of the stem regions was enriched in a small subset of the polysomal mRNA population, probably because of developmentally regulated gene expression. Exposing plants to low Psi(w) for 24 hours induced a change in the relative abundance of a small number of polysomal mRNAs in the elongating and mature tissues, but not in the dividing tissue. After 24 to 72 hours at low Psi(w), the changes in polysomal mRNA population were reversed in the elongating tissue. The data indicate that changes in stem growth at low water potential are associated with changes in polysome status and polysomal mRNA in the elongating tissue.


Periorbital cyst with bone defect in a dog.
A 4-year-old female Miniature Dachshund was referred with a chief complaint of right periorbital swelling that had not responded to antibiotic therapy. Ultrasonography and fine-needle aspiration revealed that the periorbital lesion had a cystic structure without any inflammatory or neoplastic cells. Computed tomography (CT) showed that the cyst occupied a defect in the periorbital maxillary, lacrimal, and frontal bones and had invaded the nasal cavity. The lesion was histologically suspected by incisional biopsy as an epithelial cyst.


A Pilot Study on Efficacy of Lipid Bubbles for Theranostics in Dogs with Tumors.
The combined administration of microbubbles and ultrasound (US) is a promising strategy for theranostics, i.e., a combination of therapeutics and diagnostics. Lipid bubbles (LBs), which are experimental theranostic microbubbles, have demonstrated efficacy in vitro and in vivo for both contrast imaging and drug delivery in combination with US irradiation. To evaluate the clinical efficacy of LBs in combination with US in large animals, we performed a series of experiments, including clinical studies in dogs. First, contrast-enhanced ultrasonography using LBs (LB-CEUS) was performed on the livers of six healthy Beagles. The hepatic portal vein and liver tissue were enhanced; no adverse reactions were observed. Second, LB-CEUS was applied clinically to 21 dogs with focal liver lesions. The sensitivity and specificity were 100.0% and 83.3%, respectively. These results suggested that LB-CEUS could be used safely for diagnosis, with high accuracy. Finally, LBs were administered in combination with therapeutic US to three dogs with an anatomically unresectable solid tumor in the perianal and cervical region to determine the enhancement of the chemotherapeutic effect of liposomal doxorubicin; a notable reduction in tumor volume was observed. These findings indicate that LBs have potential for both therapeutic and diagnostic applications in dogs in combination with US irradiation.


Effectiveness of Counseling on Chronic Pain Management in Patients with Temporomandibular Disorders.
To evaluate the effectiveness of counseling on pain intensity and oral health-related quality of life (OHRQoL) in temporomandibular disorders (TMD) patients. Fifty female patients diagnosed with TMD were divided into two groups: a group of waiting list patients (control group) and a group of patients who received counseling therapy (experimental group) involving education about etiologic factors, avoidance of parafunctional habits, and sleep, as well as dietary advice. All patients were evaluated at baseline and 7, 15, 30, and 60 days later. Patients reported pain intensity using a visual analog scale (VAS), and the Oral Health Impact Profile (OHIP-14) was used to assess the impact of pain on OHRQoL. Statistical analyses were performed using the split-plot analysis of variance (SPANOVA) design, with post hoc Student t tests for independent samples and for dependent samples, adopting a significance level of P < .05. The control group consisted of 24 female patients with a mean age of 39.96 ± 13.93 years, and the experimental group consisted of 26 female patients with a mean age of 35.15 ± 10.78 years. Counseling was considered effective for reducing pain intensity, with a significant improvement observed at 7 days (P < .001). Counseling was also responsible for a significant improvement in the impact of TMD on OHRQoL at all follow-up time points analyzed (P < .001). When comparing the groups, a significant difference was observed for both pain intensity and TMD impact on OHRQoL during follow-up (P < .05). Counseling seems to significantly improve pain and OHRQoL in patients.


Source reduction in an anthrax-contaminated mail facility.
Following the 2001 bioterrorist attacks using letters containing Bacillus anthracis spores, a number of sites had to be decontaminated. Source reduction-the process of reducing biocontamination levels prior to the main decontamination treatment-is a critical step in decontaminating buildings. Such activities include the removal of items from a facility either for off-site treatment and reuse or ultimate disposal, as well as the pretreatment of certain surfaces in the facility. In addition, waste disposal activities are initiated during the source reduction process and continued throughout the overall remediation process. This article presents the source reduction and waste disposal activities at the Department of State (DoS) Diplomatic Pouch and Mail Facility (SA-32) in Loudoun County, Virginia. At this facility, nearly all materials were removed before the fumigations with vaporized hydrogen peroxide. The materials that were removed from the site were divided into essential and nonessential items, and nonessential items were further categorized as recyclable or not recyclable. Removed materials were subjected to 5 different decontamination technologies, either before or after removal. The surfaces of large, fixed items and interior facility structures were pretreated and then fumigated. Environmental sampling was performed throughout the source reduction process for quality assurance purposes. The source reduction process lasted about 9 months and cost about $4.3 million. The overall remediation process took about 16 months and cost $8.6 million.


Localization of the sheep FcRn in the mammary gland.
Among the multiple functions, which have been identified for the neonatal Fc receptor (FcRn), we study its role in the IgG transport in the mammary gland during the colostrum formation. For this reason, we have obtained several mammary gland biopsies from a pregnant sheep around parturition. The presence of the FcRn heavy chain mRNA was detected exclusively in the acinar and ductal epithelial cell by in situ hybridization (ISH). We detected strong signal in samples harvested 24 and 10 days prepartum; however, in samples we collected postpartum was barely detectable. Immunohistochemistry confirmed our ISH data. The cytoplasm of the epithelial cells of the acini and ducts in the mammary gland biopsies stained homogeneously before parturition, although a remarkable difference was observed in the pattern after lambing. The signal indicated uneven distribution of the FcRn alpha chain in the epithelial cells 1 and 5 days postpartum, since the apical sides of the epithelial cells were highlighted. The presence of the FcRn in the acinar and ductal epithelial cells and the obvious change of its distribution before and after parturition suggest that FcRn plays an important role in the IgG transport during colostrum formation. FcRn expression was also found in the lamb duodenal crypt epithelial cells, which have been previously demonstrated to secrete IgG1 in newborn ruminants, suggesting secretory role of the FcRn in ruminant epithelial cells.


Transcriptome-Wide N6-Methyladenosine (m6A) Methylome Profiling of Heat Stress in Pak-choi (Brassica rapa ssp. chinensis).
Background: In higher eukaryotes, N6-methyladenosine (m6A) is the most common internal form of messenger RNA modification. By mapping the m6A methyl genome in multiple species, the potential regulatory function of reversible m6A methylation on mRNA is revealed. Recent studies have shown that RNA m6A modification influences mRNA transcription, location, translation, stability, splicing, and nuclear export. However, there are not enough data on the m6A transcriptome-wide map and its potential biological role in the heat stress of Pak-choi (Brassica rapa ssp. chinensis). Methods: In this work, MeRIP-seq was used to obtain the first transcriptome-wide profiling of RNA m6A modification in Pak-choi. Meanwhile, the transcriptome data were obtained by analyzing the input samples' sequencing data. Results: Our research indicated that with three replicates, there were 11,252 common m6A peaks and 9729 common m6A-containing genes identified in the normal (CK) and heat stress (T43) groups. It was found that m6A peaks were highly enriched in the 3' untranslated region in both CK and T43 groups. About 80% of the genes have one m6A site. The consensus sequence of m6A peaks was also enriched, which showed as AAACCV (V: U/A/G). In addition, association analysis found that there is a certain relationship between the degree of m6A methylation and the transcription level, indicating that m6A plays a certain regulatory role in gene expression. Conclusion: This comprehensive map in the study may provide a solid basis for determining the potential function of RNA m6A modification in Pak-choi under normal (CK) and heat stress (T43) conditions.


[Microcirculatory aspects of the pathogenesis of diffuse cardiosclerosis in chronic ischemic heart disease].
The morphogenesis of the diffuse cardiosclerosis in chronic coronary heart disease (CHD) in connection with the microcirculatory alterations has been studied. 16 cardiobiopsies taken during the aortocoronary shunt operations and the heart of 15 rabbits fed atherogenic diet (0.25 g/kg cholesterol) for 8 weeks and at the same time given pituitrin every day at the dose level of 0.2 U/kg were examined. It is established that disorganization of the transporttrophic tissue supply stimulates desmoplastic processes. Protein-glycosaminoglycane complexes that are difficult to eliminate are formed in the interstitium with the participation of stromal cells; these complexes fill the loop-like constructions of the basic amorphous substance, are precipitated on the fibrillar structures influencing the permeability of stroma and its neoformation. Pathological changes in the interstitium structure are brought about in spite of the desmolithic reaction in which an express and lasting components may be distinguished. The most important element of diffuse cardiosclerosis in CHD is "vasophilic" orientation of fibroblasts leading to the formation of an additional barrier immediately arount blood capillaries.


Adeno-Associated Virus Production, Purification, and Titering.
Adeno-associated virus (AAV) vectors are exemplary tools for studying gene function in vivo and are particularly favorable for transferring genes of interest into brain tissues. They have shown great promise as a gene therapy vector for preclinical and clinical applications. However, the ability to use this tool is often hampered because the viruses themselves are not readily available. Many methods have been developed for AAV production. Here, we describe a simple method for small- to medium-scale (1012 -1013 viral particles) production of AAV based on Polyethylenimine Max (PEI Max)-mediated triple transfection of HEK 293 cells and purification with iodixanol gradient ultracentrifugation. These methods will provide users with ample material of sufficient quality for performing in vivo gene transfer. © 2018 by John Wiley & Sons, Inc.


5-HT1A receptors modulate small-conductance Ca2+-activated K+ channels.
Small-conductance calcium-activated potassium channels (SK) are responsible for the medium afterhyperpolarisation (mAHP) following action potentials in neurons. Here we tested the ability of serotonin (5-HT) to modulate the activity of SK channels by coexpressing 5-HT1A receptors with different subtypes of SK channels (SK1, SK2, and SK3) in Xenopus laevis oocytes. SK channels were activated by intracellular injection of Cd2+. Subsequent activation of 5-HT1A receptors by 8-OH-DPAT always produced an inhibition of the SK current, showing the existence of a specific pathway between the receptor and the ion channel. To investigate the physiological relevance of this pathway, we characterized the mAHP present after action potentials in spinal motoneurons recorded in a slice preparation from the lumbar spinal cord of the adult turtle. By performing current and voltage clamp recordings, we showed that 8-OH-DPAT specifically inhibited the fraction of the AHP mediated by SK channels. We conclude that the activity of SK channels is modulated by activation of serotonergic receptors.


Development and evaluation of a nucleic acid sequence based amplification (NASBA) protocol for the detection of enterovirus RNA in cerebrospinal fluid samples.
A nucleic acid sequence based (NASBA) assay for the generic detection of enterovirus RNA in cerebrospinal fluid (CSF) samples was developed and compared with an established reverse transcription/nested polymerase chain reaction (PCR) protocol. The sensitivity of NASBA followed by detection of amplicons with a biotinylated oligonucleotide probe was < or = ten copies of enterovirus RNA, indicating that enterovirus NASBA achieves a similar sensitivity as nested PCR. Moreover, NASBA detected a panel of 22 different serotypes of the species poliovirus, human enterovirus A, human enterovirus B and human enterovirus C completely. For evaluating NASBA as a diagnostic tool, 61 CSF samples of patients suffering from aseptic meningitis were tested in parallel with NASBA and nested PCR. NASBA detected enterovirus RNA in four CSF samples, two of these were also positive by nested PCR and two other CSF samples were positive only by nested PCR (in total six positive samples). All other 55 of 61 CSF samples were concordantly enterovirus negative by both methods. In conclusion, the more simple to handle 'one step' NASBA is as sensitive as nested PCR and may be used as an alternative method for the detection of enterovirus RNA in CSF samples. Enterovirus NASBA is a 'one step' RNA amplification procedure that is less prone to cross-contamination compared to a three step nested PCR.


Temporal expression of respiratory genes in an enrichment culture containing Dehalococcoides ethenogenes.
Multiple reductive dehalogenase (RDase), hydrogenase (H2ase), and other respiration-associated (RA) oxidoreductase genes have been identified in cultured representatives of Dehalococcoides. Although their products are likely to play key roles in the environmentally important process of reductive dechlorination, very little information is available about their regulation and specific functions. Here we show increased expression and temporal variability in the expression of five RDase genes and in the expression of genes for a putative formate dehydrogenase (Fdh) and two H2ases, including a periplasmic [Ni/Fe] H2ase (Hup) and a cytoplasmic [Fe] H2ase (Vhu). mRNA transcripts extracted from tetrachloroethene-dechlorinating mixed cultures corresponding to Fdh, the H2ase Hup, and the RDase targets TceA and DET0162 were expressed most highly, with average levels 34 (+/- 7.5)-, 23 (+/- 6.7)-, 16 (+/- 3.3)-, and 13 (+/- 3.3)-fold higher, respectively, than that for RNA polymerase (RpoB). H2ase and RA transcripts reached their respective expression maxima within the first 2 h after feeding. RDase transcripts, however, were most highly expressed after 3 h and exhibited greater temporal variability than other transcripts. Comparison with D. ethenogenes strain 195 pure culture expression levels indicated that RDase DET1545 was more highly expressed in mixed cultures, where, on average, its transcript level was sixfold higher than that of RpoB. While the specific functions of several of these gene products remain elusive, the high expression levels and temporal variability reported here suggest that these groups of enzymes are metabolically important for the respiration of chlorinated ethenes in mixed cultures containing Dehalococcoides.


Orthophosphate control of glucose-6-phosphate dehydrogenase light modulation in relation to the induction phase of chloroplast photosynthesis.
High concentrations of orthophosphate (Pi) inhibited CO(2)-dependent O(2) evolution and prevented the inactivation of glucose-6-P dehydrogenase by light in intact spinach and barley chloroplasts. Addition of glycerate-3-P to chloroplasts inhibited by Pi in the light, induced O(2) evolution and caused rapid inactivation of glucose-6-P dehydrogenase. The activity of phosphofructokinase detected in chloroplast preparations was not affected by light or by Pi.Dihydroxyacetone-P was a major product of chloroplast photosynthesis when optimum concentrations of Pi were used. Chloroplasts continued to form dihydroxyacetone-P at a slow rate in the presence of Pi at concentrations (2 to 4 millimolar) that gave complete inhibition of CO(2)-dependent O(2) evolution. Formation of dihydroxyacetone-P in the presence of 4 millimolar Pi was stimulated by light and either O(2) (150 micromolar) or sparker amounts of oxaloacetate.Conditions that favored dihydroxyacetone-P formation (high O(2) or low O(2) plus oxaloacetate) increased the optimum Pi concentration for CO(2)-dependent O(2) evolution and stimulated O(2) evolution at high concentrations of Pi. The stimulation of O(2) evolution at superoptimal concentrations of Pi by O(2) or oxaloacetate was prevented by dithiothreitol.The results suggested that formation of pentose-P pathway intermediates via the oxidative pentose-P pathway may be limited by availability of NADP in the light but may occur at significant rates and thereby contribute to termination of the induction phase of O(2) evolution.


Structural trends in ten-vertex endohedral clusters, M@E(10) and the synthesis of a new member of the family, [Fe@Sn10]3-.
The synthesis of a new endohedral ten-vertex Zintl ion cluster, [Fe@Sn10](3-), isoelectronic with [Fe@Ge10](3-), is reported. In an attempt to place this new cluster within the context of the known structural chemistry of the M@E10 family (M = transition metal, E = main group element), we have carried out a detailed electronic structure analysis of the different structural types: viz bicapped square antiprismatic ([Ni@Pb10](2-), [Zn@In10](8-)), tetra-capped trigonal prismatic ([Ni@In10](10-)) and the remarkable pentagonal prismatic [Fe@Ge10](3-) and [Co@Ge10](3-). We establish that the structural trends can be interpreted in terms of a continuum of effective electron counts at the E10 cage, ranging from electron deficient (<4n + 2) in [Ni@In10](10-) to highly electron rich (>4n + 2) in [Fe@Ge10](3-). The effective electron count differs from the total valence electron count in that it factors in the increasingly active role of the metal d electrons towards the left of the transition series. The preference for a pentagonal prismatic geometry in [Fe@Ge10](3-) emerges as a natural consequence of backbonding to the cage from four orthogonal 3d orbitals of the low-valent metal ion. Our calculations suggest that the new [Fe@Sn10](3-) cluster should also exhibit structural consequences of backbonding from the metal to the cage, albeit to a less extreme degree than in its Ge analogue. The global minimum lies on a very flat surface connecting D4d, C2v and C3v-symmetric minima, suggesting a very plastic structure that may be easily deformed by the surrounding crystal environment. If so, then this provides a new and quite distinct structural type for the M@E10 family.


Morphology of human vestibular ganglion.
Vestibular ganglion cells of mammals have been considered to be myelinated like spiral ganglion cells. Recently, electron microscopic studies reported that the ganglion cells of human vestibular ganglion were mostly unmyelinated unlike in other mammals. The purpose of this study was to find the myelin sheaths of human vestibular ganglion cells by light and laser microscopy.


Minimally invasive robotic breast reconstruction surgery.
The rising popularity of robotic surgery has enabled surgeons to continue to expand the uses of robotic surgery. Robotic surgery offers minimally invasive approaches coupled with tremor elimination, up to seven degrees of freedom, ergonomic positioning, 3D magnified vision and improved resolution. We describe robotic surgery techniques for nipple-sparing mastectomies, latissimus dorsi muscle flap harvest, deep inferior epigastric perforator (DIEP) flap pedicle harvest, and robotic microsurgical anastomoses. By using a robotic system the surgeon is able to offer not only a minimally invasive approach to the patient but the surgeon's ability can be improved upon as well. This improved ability is best characterized in the robotic supermicrosurgical anastomosis where even the faintest surgeon's tremor is exploited. However, within the robotic system tremor is eliminated. We are now able to offer patients a completely minimally invasive approach to ablative breast surgery and breast reconstruction. A patient could have a robotic nipple-sparing mastectomy, followed by a robotic DIEP reconstruction with a robotic microsurgical anastomosis. The patient could even have robotic lymphovenous bypass to address lymphedema that could have arisen after an axillary dissection. A completely robotic surgical approach maximizes both utilization of the robotic system and patient benefit. By using robotic techniques in flap harvest the morbidity of traditional open surgeries is minimized and the use of robotic anastomoses expands the limits of human precision.


Integrating nanoscience into the classroom: perspectives on nanoscience education projects.
The National Nanotechnology Initiative has motivated substantial growth in nanoscience and nanotechnology research in the United States and beyond. One of the central goals of the National Nanotechnology Initiative is the development and education of future generations of nanoscience researchers. This Nano Focus provides a brief history of nanoscience education, including curricula that have been implemented successfully into secondary and college institutions, as described in the symposium "Integrating Nanoscience into the College and High School Classroom" at the 237th American Chemical Society National Meeting in March 2009.