Thyroid involvement in hepatitis C - associated mixed cryoglobulinemia.
The prevalence and clinical features of thyroid involvement in patients with hepatitis C virus-associated mixed cryoglobulinemia (MC+HCV) have been reviewed. A PubMed Medline search was conducted through December 2011 to identify all studies that reported thyroid involvement in MC+HCV patients. Reference lists of the papers initially detected were manually searched to identify additional relevant reports. Studies had to contain sufficient and clear information to be included. In MC+HCV patients, the following thyroid autoimmune abnormalities were significantly more frequent than in controls: high levels of serum anti-thyroperoxidase autoantibody (AbTPO); high levels of serum AbTPO and/or anti-thyroglobulin autoantibody; humoral and ultrasonographical signs of thyroid autoimmunity (35% vs 16%); prevalence of subclinical hypothyroidism (11% vs 2%). Also, the prevalence of papillary thyroid cancer has been found higher in MC+HCV patients than in controls, in particular in patients with autoimmune thyroiditis. The involvement of T helper 1 immunity and chemokine (C-X-C motif) ligand 10 (CXCL10) may be the pathogenetic basis of the association between MC+HCV and thyroid autoimmunity. These results show a high prevalence of thyroid disorders in patients with MC+HCV and point to the need for careful monitoring of thyroid function in these patients.


Melasma in men. A clinical and histologic study.
Melasma is characterized by a facial hypermelanosis of light to dark brown color, being more common in women of Hispanic origin. In this study, 27 men with melasma were evaluated clinically and histologically to compare their features with those of previous studies. Three patterns of localization were recognized, namely, centrofacial, malar, and mandibular. On the basis of Wood's light examination, an epidermal, a dermal, and a mixed type were identified. Epidermal hyperpigmentation only and epidermal and dermal hyperpigmentation were found in histologic analysis of the cases. Significant etiologic factors included exposure to sunlight in 66.6% as well as a familial predisposition in 70.4% of the cases. This study demonstrated that melasma in men shares the same clinicohistologic characteristics as in women, but hormonal factors do not seem to play major significant role.


BRCA1 haploinsufficiency: consequences for breast cancer.
In their study, Konishi et al. generate and study cell lines that carry one mutant and one normal copy of BRCA1. These heterozygous (noncancerous) cells are noteworthy in that they carry an increased number of copy number alterations and have diminished DNA repair capacity. These observations go beyond the classical two-hit hypothesis for inherited cancers, whereby the cell phenotype is normal until the second allele is lost through somatic mutation or epigenetic silencing. They propose that BRCA1 heterozygosity is associated with increased genomic instability, which accelerates the mutation rate of other critical genes, including the second copy of BRCA1. They conclude that BRCA1 is a haploinsufficient tumor suppressor gene, which when lost through inheritance, increases a cell's susceptibility to acquire further mutations.


5-HT2 receptor binding and 5-HT uptake in mouse brain: developmental changes and the relationship to audiogenic seizure susceptibility in DBA/2J mice.
Abnormal function of serotonergic neurones may be involved in the age-related susceptibility of DBA/2J mice to generalised convulsions induced by auditory stimulation. We have measured 5-HT2 receptor binding sites and synaptosomal 5-HT uptake in 5 brain regions of DBA/2J mice at ages before, during and after their maximal susceptibility to audiogenic seizures and in age-matched C57 B1/6 mice, a strain resistant to audiogenic seizures at all ages. The number of 5-HT2 binding sites was 20% higher in the cerebral cortex of DBA/2J than C57 B1/6 mice at the time of maximal susceptibility of DBA/2J mice to audiogenic seizures but did not differ at other ages. The number of 5-HT2 binding sites did not differ between the two strains at the ages studied in forebrain, mid-brain, hippocampus and pons-medulla. A marked reduction in the number of 5-HT2 binding sites was apparent in the mid-brain, hippocampus and pons-medulla of both strains of mice between 13-15 days of age and 21-23 days of age. Synaptosomal 5-HT uptake did not differ significantly between DBA/2J and C57 B1/6 in any of the brain regions at the ages studied. The higher density of cortical 5-HT2 binding sites in DBA/2J mice may contribute to their susceptibility to sound-induced seizures.


Therapeutic Potential of Melatonin in the Regulation of MiR-148a-3p and Angiogenic Factors in Breast Cancer.
The high mortality rate of breast cancer is related to the occurrence of metastasis, a process that is promoted by tumor angiogenesis. MicroRNAs are small molecules of noncoding mRNA that play a key role in gene regulation and are directly involved in the progression and angiogenesis of various tumor types, including breast cancer. Several miRNAs have been described as promoters or suppressors angiogenesis and may be associated with tumor growth and metastasis. Melatonin is an oncostatic agent with a capacity of modifying the expression of innumerable genes and miRNAs related to cancer. The aim of this study was to evaluate the role of melatonin and the tumor suppressor miR- 148a-3p on angiogenesis of breast cancer. MDA-MB-231 cells were treated with melatonin and modified with the overexpression of miR-148a-3p. The relative quantification in real-time of miR-148a-3p, IGF-IR and VEGF was performed by real-time PCR. The protein expression of these targets was performed by immunocytochemistry and immunohistochemistry. Survival, migration and invasion rates of tumor cells were evaluated. Finally, the xenograft model of breast cancer was performed to confirm the role of melatonin in the tumor. The melatonin was able to increase the gene level of miR-148a-3p and decreased the gene and protein expression of IGF-1R and VEGF, both in vitro and in vivo. In addition, it also had an inhibitory effect on the survival, migration and invasion of breast tumor cells. Our results confirm the role of melatonin in the regulation of miR-148a-3p and decrease of angiogenic factors.


Genetic and post-mortem mRNA analysis of the 14-3-3 genes that encode phosphoserine/threonine-binding regulatory proteins in schizophrenia and bipolar disorder.
Previous work with animal models of psychosis, human genetic studies, and human post-mortem gene expression studies implicate the 14-3-3 family of genes in schizophrenia. The 14-3-3 genes code for a family of proteins that bind to and regulate other proteins, and they modulate neurodevelopment, cell-division, signal transduction and gene transcription. To explore the role of five 14-3-3 isoforms (beta, gamma, epsilon, zeta, and eta) in schizophrenia by: (1) comparing mRNA levels in post-mortem brain from schizophrenic, bipolar and control subjects and (2) assessing genetic association with schizophrenia in both case-control and nuclear family samples. Quantitative PCR (q-PCR) was used to determine relative mRNA levels in dorsolateral prefrontal cortex (Brodmann's area 46) samples donated by the Stanley Medical Research Institute (SMRI). Selected SNPs were genotyped in all five isoforms for association analysis in both family and case-control samples. No significant differences in 14-3-3 mRNA expression levels between the diagnostic groups were found. A significant genetic association with schizophrenia was found for the 14-3-3zeta isoform in a subset of nuclear families of British ancestry (TDT: chi(2)=7.2; df=1; p=0.0073), in the case-control sample overall (p=0.011), and in a subset of the case-control sample. The results, in combination with other published evidence, suggest that further work is necessary to clarify what role the 14-3-3 genes may play in the etiology and pathogenesis of schizophrenia.


Determination of sialylated and neutral oligosaccharides in urine by mass spectrometry.
This protocol describes a method to allow for the detection of specific oligosaccharide fragments in urine by tandem mass spectrometry. The detection of fragments with specific masses indicates the presence of one of a number of diseases where the deficiency of lysosomal enzymes involved in the degradation of the glyco- moieties of glycoproteins is present in the patient. This method describes the derivatization of oligosaccharides present in urine with phenyl-1-methylpyrazolone, which renders them hydrophobic, thus allowing desalting with Combi cleanup columns prior to injection. This method allows the detection of storage of oligosaccharides, which may indicate the presence of one of the infantile Pompe disease, α-mannosidosis, Gm1-gangliosidosis, Sandhoff disease, sialidosis, galactosialidosis, I-cell disease, and aspartylglucosaminuria.


Relationship of renal threshold for bicarbonate reabsorption to urinary sodium excretion in premature infants.
Renal threshold for bicarbonate reabsorption and urinary sodium excretion of nine healthy male premature infants with a mean birth weight of 1734 g (range 1650-1900) was determined on the 7th day of life, and subsequently weekly for 6 consecutive weeks, by applying a repeated NH4Cl load. A close negative correlation was found between bicarbonate threshold and urinary sodium excretion (p less than 0.01). It is suggested that the limited renal capacity to reabsorb sodium may account for the low bicarbonate threshold in premature infants.


[Effects of Side-effect Attenuation Prescriptions on Cyclophosphamide-induced Myelosuppression in Mice].
To determine the effects of Side-effect Attenuation Prescriptions on the levels of white blood cells (WBC), hemoglobin (HGB) and platelet (PLT) in peripheral blood of mice influenced by cyclophosphamide (CTX). Mice were randomly divided into 6 groups: normal control group, myelosuppression group induced by CTX (model group), recombinant human granulocyte colony stimulating factor (G-CSF) group, and Side-effect Attenuation Prescriptions group (experimental groups with high, middle, and low dosages). Marrow depressed models were established by injecting CTX intraperitoneally (40 mg/kg, once/d, 10 d) to the mice. High, middle, and low dosage experimental groups received 40 g/kg, 20 g/kg, and 10 g/kg Side-effect Attenuation Prescriptions once a day for two weeks, respectively. Mice in the G-CSF group were given G-CSF (50 μg/kg) by hypodermic injection three days before blood sampling. Levels of WBC, HGB and PLT counts in peripheral bloods of the mice were detected at 7 d and 14 d after the marrow depressed models were established. The Side-effect Attenuation Prescriptions slowed down the decline of blood levels of WBC, HGB and PLT induced by CTX (P < 0.05), and accelerated the recovery of WBC and HGB levels compared with model group at 14 d (P < 0.05). The Side-effect Attenuation Prescriptions can accelerate recovery of WBC, HGB and PLT in peripheral bloods of mice.


CD51 distinguishes a subpopulation of bone marrow mesenchymal stem cells with distinct migratory potential: a novel cell-based strategy to treat acute myocardial infarction in mice.
Experimental and clinical trials have demonstrated the efficiency of bone marrow-derived mesenchymal stromal/stem cells (bMSCs) in the treatment of myocardial infarction. However, after intravenous injection, the ineffective migration of engrafted bMSCs to the hearts remains an obstacle, which has an undesirable impact on the efficiency of cell-based therapy. Therefore, we attempted to identify a marker that could distinguish a subpopulation of bMSCs with a promising migratory capacity. Here, CD51-negative and CD51-positive cells were isolated by flow cytometry from Ter119-CD45-CD31-bMSCs and cultured in specifically modified medium. The proliferation ability of the cells was evaluated by 5-ethynyl-2'-deoxyuridine (EdU) staining or continuously monitored during culture, and the differentiation potential was assessed by culturing the cells in the appropriate conditioned media. Wound healing assays, transwell assays and quantitative polymerase chain reaction (qPCR) were used to measure the migratory ability. The mice were subjected to a sham operation or myocardial infarction (MI) by permanently occluding the coronary artery, and green fluorescent protein (GFP)-labelled cells were transplanted into the mice via intravenous infusion immediately after MI. Heart function was measured by echocardiography; infarct myocardium tissues were detected by triphenyl tetrazolium chloride (TTC) staining. Additionally, immunofluorescence staining was used to verify the characteristics of CD51+bMSCs and inflammatory responses in vivo. Statistical comparisons were performed using a two-tailed Student's t test. In this study, the isolated CD51-bMSCs and CD51+bMSCs, especially the CD51+ cells, presented a favourable proliferative capacity and could differentiate into adipocytes, osteocytes and chondrocytes in vitro. After the cells were transplanted into the MI mice by intravenous injection, the therapeutic efficiency of CD51+bMSCs in improving left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) was better than that of CD51-bMSCs. Compared with CD51-bMSCs, CD51+bMSCs preferentially migrated to and were retained in the infarcted hearts at 48 h and 8 days after intravenous injection. Accordingly, the migratory capacity of CD51+bMSCs exceeded that of CD51-bMSCs in vitro, and the former cells expressed higher levels of chemokine receptors or ligands. Interestingly, the retained CD51+bMSCs retained in the myocardium possessed proliferative potential but only differentiated into endothelial cells, smooth muscle cells, fibroblasts or cardiomyocytes. Transplantation of CD51+bMSCs partially attenuated the inflammatory response in the hearts after MI, while the potential for inflammatory suppression was low in CD51-bMSC-treated mice. These findings indicated that the CD51-distinguished subpopulation of bMSCs facilitated proliferation and migration both in vitro and in vivo, which provided a novel cell-based strategy to treat acute MI in mice by intravenous injection.


Ovarian fibromatosis and sotos syndrome with a new genetic mutation.
Sotos syndrome is one the most common overgrowth conditions, after Beckwith-Wiedemann syndrome. As with other overgrowth syndromes, Sotos syndrome can be associated with an increased risk of tumors. We describe a young girl with Sotos syndrome and ovarian fibromatosis with a new mutation not reported before in the literature. Development of ovarian tumor in Sotos syndrome has been poorly documented. Ovarian fibromatosis is a very rare non neoplastic disease. Management is guided by the benignity of the lesion and consists of surgical excision of the fibroma.


Structural and functional characterization of the deep-sea thermophilic bacteriophage GVE2 tailspike protein.
The genome of the thermophilic bacteriophage GVE2 encodes a putative tailspike protein (GVE2 TSP). Here we report the crystal structure of the truncated GVE2 TSP at 2.0-Å resolution lacking 204 amino acid residues at its N-terminus (ΔnGVE2 TSP), possessing a "vase" outline similar to other TSP's structures. However, ΔnGVE2 TSP displays structural characteristics distinct from other TSPs. Despite lacking 204 amino acid residues, the head domain forms an asymmetric trimer compared to symmetric in other TSPs, suggesting that its long N-terminus may be unique to the long-tailed bacteriophages. Furthermore, the α-helix of the neck is 5-7 amino acids longer than that of other TSPs. The most striking feature is that its binding domain consists of a β-helix with 10 turns, whereas other TSPs have 13 turns, even including the phage Sf6 TSP, which is the closest homologue of GVE2 TSP. The C-terminal structure is also quite different with those of other TSPs. Furthermore, we observed that ΔnGVE2 TSP can slow down growth of its host, demonstrating that this TSP is essential for the phage GVE2 to infect its host. Overall, the structural characteristics suggest that GVE2 TSP may be more primitive than other phage TSPs.


Safety, immunogenicity and protective efficacy in mice of a new cell-cultured Lister smallpox vaccine candidate.
It is now difficult to manufacture the first-generation smallpox vaccine, as the process could not comply with current safety and manufacturing regulations. In this study, a candidate non-clonal second-generation smallpox vaccine developed by Sanofi-Pasteur from the Lister strain has been assessed using a cowpox virus challenge in mice. We have observed similar safety, immunogenicity and protection (from disease and death) after a short or long interval following vaccination, as well as similar virus clearance post-challenge, with the second-generation smallpox vaccine candidate as compared to the traditional vaccine used as a benchmark.


Comparison of capillary electrophoresis-based immunoassay with fluorescence polarization immunoassay for the immunodetermination of methamphetamine using various methamphetamine antibodies.
An accurate and simple immunoassay using capillary electrophoresis (CE) with laser-induced fluorescence (LIF) was performed for the detection of methamphetamine (MA) in urine. The CE-LIF was conducted with an untreated fused-silica column using antiserum and a tracer of fluorescein isothiocyanate (FITC)-labeled MA. This CE-LIF system was compared with fluorescence polarization immunoassay (FPIA) in a TDx analyzer in the photo-check mode using the same FITC-labeled tracer and the same antiserum. Various antibodies, not only those prepared by our own immunogens but also those from commercial sources, were screened and characterized in both assay systems with regard to sensitivity, precision, and cross-reactivity. Both systems satisfied analytical precision and gave similar cross-reactivity patterns. However, the CE-LIF-based immunoassay was approximately one order superior to FPIA in sensitivity, requiring less volume of sample, antiserum, and tracer for the assay. Considering that the FPIA system is well known to be a useful tool for screening antibodies and detecting drugs, the CE-LIF-based immunoassay system, which is seemingly more advantageous than the FPIA system, appears to have great power for the characterization of antibodies and for the detection of MA in urine.


Reassortment compatibility between PB1, PB2, and HA genes of the two influenza B virus lineages in mammalian cells.
In addition to influenza A subtypes, two distinct lineages of influenza B virus also cause seasonal epidemics to humans. Recently, Dudas et al. have done evolutionary analyses of reassortment patterns of the virus and suggested genetic lineage relationship between PB1, PB2, and HA genes. Using genetic plasmids and reassortant viruses, we here demonstrate that a homologous lineage PB1-PB2 pair exhibits better compatibility than a heterologous one and that the lineage relationship between PB1 and HA is more important for viral replication than that between PB2 and HA. However, co-adaptation of PB1-PB2-HA genes appears to be affected by complete gene constellation.


Community point distribution of insecticide-treated bed nets and community health worker hang-up visits in rural Zambia: a decision-focused evaluation.
In 2013, the Zambian Ministry of Health through its National Malaria Control Programme distributed over two million insecticide-treated bed nets (ITNs) in four provinces using a door-to-door distribution strategy, and more than 6 million ITNs were allocated to be distributed in 2014. This study was commissioned to measure attendance rates at a community point distribution and to examine the impact of follow-up community health worker (CHW) hang-up visits on short and medium-term ITN retention and usage with a view of informing optimal ITN distribution strategy in Zambia. Households received ITNs at community point distributions conducted in three rural communities in Rufunsa District, Zambia. Households were then randomly allocated into five groups to receive CHW visits to hang any unhung ITNs at different intervals: 1-3, 5-7, 10-12, 15-17 days, and no hang-up visit. Follow-up surveys were conducted among all households at 7-11 weeks after distribution and at 5-6 months after distribution to measure short- and medium-term household retention and usage of ITNs. Of the 560 pre-registered households, 540 (96.4 %) attended the community point distribution. Self-installation of ITNs by households increased over the first 10 days after the community point distribution. Retention levels remained high over time with 90.2 % of distributed ITNs still in the household at 7-11 weeks and 85.7 % at 5-6 months. Retention did not differ between households that received a CHW visit and those that did not. At 7-11 weeks, households had an average of 73.8 % of sleeping spaces covered compared to 80.3 % at 5-6 months. On average, 65.6 % of distributed ITNs were hanging at 7-11 weeks compared to 63.1 % at 5-6 months. While a CHW hang-up visit was associated with increased usage at 7-11 weeks, this difference was no longer apparent at 5-6 months. This evaluation revealed that (1) the community point distributions achieved high attendance rates followed by acceptable rates of short-term and medium-term ITN retention and usage, as compared to reported rates achieved by door-to-door distributions in the recent past, (2) CHW hang-up visits had a modest short-term impact on ITN usage but no medium-term effect, and (3) community point distributions can yield sizeable time savings compared to door-to-door distributions.


Thrombotic microangiopathy induced by interferon beta in patients with multiple sclerosis: three cases treated with eculizumab.
Interferon-beta (IFN-beta) is one of the most widely prescribed medications for relapsing-remitting multiple sclerosis (RRMS). IFN-related thrombotic microangiopathy (TMA) is a rare but severe complication, with a fulminant clinical onset and a possibly life-threatening outcome that may occur years after a well-tolerated treatment with IFN. Most patients evolve rapidly to advanced chronic kidney disease and eventually to renal failure. We performed a retrospective analysis of TMA cases diagnosed and managed in our Nephrology Department from 2010 to 2015, and performed a literature review of IFN-beta-induced TMA. Three cases of TMA among patients treated with IFN-beta were identified who did not show any renal improvement following conventional therapy: IFN withdrawal and plasma exchange (PE, range 8-18) sessions. All of them responded favourably to eculizumab, with progressive clinical and renal improvement, allowing dialysis discontinuation, without recurrence of TMA during a long-term follow-up (range 1-5 years). TMA is a recognized severe complication in RRMS patients treated with IFN-beta. Withdrawal of IFN and treatment with PE, steroids or rituximab did not improve the poor renal prognosis in our three patients and in all the previously described cases in the literature. In our experience, eculizumab had a strikingly favourable effect on renal recovery, suggesting a role of IFN-beta as a trigger in complement-mediated TMA. Neurologists and nephrologists should be vigilant to this complication to prevent possibly irreversible renal damage.


A dynamic interplay of enhancer elements regulates Klf4 expression in naïve pluripotency.
Transcription factor (TF)-directed enhanceosome assembly constitutes a fundamental regulatory mechanism driving spatiotemporal gene expression programs during animal development. Despite decades of study, we know little about the dynamics or order of events animating TF assembly at cis-regulatory elements in living cells and the long-range molecular "dialog" between enhancers and promoters. Here, combining genetic, genomic, and imaging approaches, we characterize a complex long-range enhancer cluster governing Krüppel-like factor 4 (Klf4) expression in naïve pluripotency. Genome editing by CRISPR/Cas9 revealed that OCT4 and SOX2 safeguard an accessible chromatin neighborhood to assist the binding of other TFs/cofactors to the enhancer. Single-molecule live-cell imaging uncovered that two naïve pluripotency TFs, STAT3 and ESRRB, interrogate chromatin in a highly dynamic manner, in which SOX2 promotes ESRRB target search and chromatin-binding dynamics through a direct protein-tethering mechanism. Together, our results support a highly dynamic yet intrinsically ordered enhanceosome assembly to maintain the finely balanced transcription program underlying naïve pluripotency.


Reconstructed glycosylase base editors GBE2.0 with enhanced C-to-G base editing efficiency and purity.
Base editing techniques were developed for precise base conversion on cellular genomic DNA, which has great potential for the treatment of human genetic diseases. The glycosylase base editor (GBE) recently developed in our lab was used to perform C-to-G transversions in mammalian cells. To improve the application prospects of GBE, it is necessary to further increase its performance. With this aim, we replaced the human Ung in GBE with Ung1 from Saccharomyces cerevisiae. The resulting editor APOBEC-nCas9-Ung1 was tested at 17 chromosomal loci and was found to have an increased C-to-G editing efficiency ranging from 2.63% to 52.3%, with an average of 23.48%, which was a significant improvement over GBE, with an average efficiency of 15.54%, but with a decreased purity. For further improvement, we constructed APOBEC(R33A)-nCas9-Rad51-Ung1 with two beneficial modifications adapted from previous reports. This base editor was able to achieve even higher editing efficiency ranging from 8.70% to 72.1%, averaging 30.88%, while also exhibiting high C-to-G purity ranging from 35.57% to 92.92%, and was designated GBE2.0. GBE2.0 provides high C-to-G editing efficiency and purity in mammalian cells, making it a powerful genetic tool for scientific research or potential genetic therapies for disease-causing G/C mutations.


A model of dopamine regulation of glutamatergic synapse on medium size spiny neurons.
Spiny neurons of striatum receive glutamatergic synapses on dendritic spines on the neck of which project dopaminergic synapses. Dopamine modulates, by D1 type receptors, the glutamatergic synapses by inducing the phosphorylation of AMPA and NMDA receptors which produces an increased amplitude response. Herein we present a model where, in addition to phosphorylation, the direct modulation by dopamine of the spine resistance can cooperate in producing the observed effect on some of these synapses.


[Questionnaire to assess behavior, knowledge and attitudes on hand hygiene among physicians].
Valid and reliable instruments to measure aspects of hand hygiene are needed to determine the current situation and impact of training among health professionals. The main objective of this study was to describe the development of a questionnaire on hand hygiene among health professionals and to analyze the properties of this instrument. A questionnaire was designed and implemented between January 2010 and March 2011 at the Hospital Clínico San Cecilio (Granada, Spain). The final questionnaire consisted of 44 items that assessed provider behavior before and after contact with the patient, declarative knowledge, and attitudes to hand hygiene. The questionnaire was administered to 113 health professionals. A factor analysis was performed. Data were obtained that supported the unidimensionality of the instrument with a general convergence value that explained 39.289% of the total variance and a Cronbach's alpha of 0.784 for the established elements. Significant differences were found in hand hygiene behavior before and after contact with the patient (t=-8,991, p <0.001). Declarative knowledge and attitudes significantly predicted behavior. The questionnaire shows high internal consistency, reliability, and validity and is thus a valid tool to assess behavior, knowledge and attitudes related to hand hygiene in health professionals. This instrument also detects deficiencies in basic knowledge.


Incompressible Biventricular Model Construction and Heart Segmentation of 4D Tagged MRI.
Most automated methods for cardiac segmentation are not directly applicable to tagged MRI (tMRI) because they do not handle all of the analysis challenges: tags obscure heart boundaries, low contrast, image artifacts, and radial image planes. Other methods do not process all acquired tMRI data or do not ensure tissue incompressibility. In this chapter, we present a cardiac segmentation method for tMRI which requires no user input, suppresses image artifacts, extracts heart features using 3D grayscale morphology, and constructs a biventricular model from the data that ensures the near incompressibility of heart tissue. We project landmarks of 3D features along curves in the solution to a PDE, and embed biomechanical constraints using the finite element method. Testing on normal and diseased subjects yields an RMS segmentation accuracy of ~2 mm, comparing favorably with manual segmentation, interexpert variability and segmentation methods for nontagged cine MRI.


Contamination problem with sentinel node localization procedure: a case study.
Lymphoscintigraphy for sentinel node (SN) localization was performed on a 60-y-old man with a melanoma on his back. Skin contamination occurred as a result of the radiopharmaceutical dose administration. Skin contamination could result in a misinterpretation of the SN location. Careful observation of the procedure avoided a misinterpretation with this study.


Clinical characteristics of pulmonary hypertension in patients with pleuroparenchymal fibroelastosis.
This study aimed to investigate the clinical characteristics and prognosis of patients with pleuroparenchymal fibroelastosis (PPFE) and pulmonary hypertension (PH). We retrospectively analyzed the data of patients who were diagnosed with PPFE and underwent transthoracic echocardiography (TTE) for the evaluation of their right heart systems within 3 months of their first visit between 2011 and 2018. Patients were divided into the PH and non-PH groups based on their peak tricuspid regurgitation velocity (TRV) on TTE (cutoff, 2.8 m/s). The clinical characteristics of PH and association between PH and survival among patients with PPFE were investigated. In total, 83 patients were enrolled. Sixteen (19.3%) patients were included in the PH group. The PH group had a lower body mass index, percent predicted forced vital capacity (FVC), 6-min walk distance, and partial pressure of arterial oxygen than the non-PH group. There was no significant difference in the presence of usual interstitial pneumonia patterns in the lower lobes between the two groups. The survival period was significantly shorter in the PH group than in the non-PH group (median survival 16.3 versus 50.2 months, log-rank p < 0.001). The multivariate Cox proportional hazard model showed that male sex (hazard ratio [HR] = 4.83, p < 0.001), Krebs von den Lungen-6 (KL-6) > 550 U/mL (HR = 3.48, p = 0.005), %FVC < 50% (HR = 3.04, p = 0.028), and peak TRV > 2.8 m/s (HR = 3.26, p = 0.038) were independently associated with poor survival. PH was not rare in patients with PPFE. Male sex, increased KL-6, lower FVC, and PH were independently associated with poor survival in patients with PPFE.


A study of conformational restraints on reactivity of human PR3-specific autoantibodies (ANCA) facilitated through protein folding manipulations of a new recombinant proteinase 3 protein.
Proteinase 3 (PR3)-specific antineutrophil cytoplasmic autoantibodies (PR3-ANCA) recognize conformational epitopes on PR3. This study evaluates PR3-ANCA target epitopes utilizing a novel recombinant PR3 (rPR3) produced to accommodate manipulations of the N-terminal domain. The rPR3 molecule contains an N-terminus six histidine tag, which can be removed by enterokinase (EK) cleavage of an adjacent EK cleavage site. Once cleaved the remaining amino acids correspond to the mature N-terminus of PR3. This rPR3 can be manipulated to produce three variant forms: tagged rPR3(+his), EK-cleaved (his-tag removed) rPR3(- his), and EK-cleaved, denatured/refolded rPR3(- his/dr) (the proteolytically active form). Patients with clinically positive PR3-ANCA titers (n = 40) were confirmed for reactivity against purchased native PR3 in our system. Controls included 29 healthy volunteers and 34 MPO-ANCA patients. All PR3-ANCA sera samples tested were reactive with one or more forms of the recombinant protein (greater than mean ELISA OD 405 + 2 SDs of controls). Of significance, three sera were reactive with non-active forms only and three others were more reactive with rPR3(- his/dr) than with native PR3. The results of our evaluation of PR3-ANCA sera for reactivity against the three forms of our rPR3 protein uniquely exemplify the diverse array of epitopes within the PR3-ANCA population. This new recombinant form of PR3 should provide a suitable approach to mapping ANCA epitopes using site-directed mutagenesis.


The Impact of Burst Motor Cortex Stimulation on Cardiovascular Autonomic Modulation in Chronic Pain: A Feasibility Study for a New Approach to Objectively Monitor Therapeutic Effects.
Chronic refractory pain of various origin occurs in 30-45% of pain patients, and a considerable proportion remains resistant to pharmacological and behavioral therapies, requiring adjunctive neurostimulation therapies. Chronic pain is known to stimulate sympathetic outflow, yet the impact of burst motor cortex stimulation (burstMCS) on objectifiable autonomic cardiovascular parameters in chronic pain remains largely unknown. In three patients with chronic pain (2 facial pain/1 post-stroke pain), we compared pain intensity using a visual analog scale (VAS 1-10) and parameters of autonomic cardiovascular modulation at supine rest, during parasympathetic challenge with six cycles per minute of metronomic deep breathing, and during sympathetic challenge (active standing) at baseline and after 4 months of burstMCS compared to age-/gender-matched healthy controls. While two out of three patients were responsive after 4 months of adjunctive burstMCS (defined as pain reduction of > 30%), no differences were found in any of the three patients regarding the R-R intervals of adjacent QRS complexes (RRI, 642 vs. 676 ms) and blood pressure (BP, 139/88 vs. 141/90 mmHg). Under resting conditions, parameters of parasympathetic tone [normalized units of high-frequency oscillations of RRI (RRI-HFnu power) 0.24 vs. 0.38, root-mean-square differences of successive RRI (RRI-RMSSD) 7.7 vs. 14.7 ms], total autonomic cardiac modulation [RRI total power 129.3 vs. 406.2 ms2, standard deviation of RRI (RRI-SD) 11.6 vs. 18.5 ms, coefficient of variation of RRI (RRI-CV) 1.9 vs. 3.7%], and baroreceptor reflex sensitivity (BRS, 1.9 vs. 2.3 ms/mmHg) increased, and parameters of sympathetic tone [normalized units of low-frequency oscillations of RRI (RRI-LFnu power) 0.76 vs. 0.62] and sympatho-vagal balance [ratio of RR-LF to RRI-HF power (RRI-LF/HF ratio) 3.4 vs. 1.9] decreased after 4 months of burstMCS. Low-frequency oscillations of systolic blood pressure (SBP-LF power), a parameter of sympathetic cardiovascular modulation, increased slightly (17.6 vs. 20.4 mmHg2). During parasympathetic stimulation, the expiratory-inspiratory ratio (E/I ratio) increased slightly, while upon sympathetic stimulation, the ratio between the shortest RRI around the 15th heartbeat and the longest RRI around the 30th heartbeat after standing up (RRI 30/15 ratio) remained unchanged. Four months of adjunctive burstMCS was associated with an increase in parameters reflecting both total and parasympathetic autonomic modulation and baroreceptor reflex sensitivity. In contrast, sympathetic tone declined in our three patients, suggesting stimulation-associated improvement not only in subjectively perceived VAS pain scores, but also in objectifiable parameters of autonomic cardiovascular modulation.


Blood pressure differences between arms and association of dominant hands with blood pressure differences and carotid atherosclerosis.
Guidelines for the management of hypertension recommend that blood pressure (BP) should be measured twice at every visit; it should be measured in both arms at the first visit, and the right arm BP or higher BP should be recorded. Manufacturers of home BP monitors tend to design the device for measurement of left arm BP. The arm preferred for BP measurement differs according to the methods recommended by the society and according to the home BP monitors. The BP difference (ΔBP) is calculated by subtracting left arm BP from right arm BP. Here, we aim to first investigate which hand will give the most accurate reading by a sphygmomanometer in daily medical practice. Second, we wish to assess the association of the dominant hand with absolute BP difference (|ΔBP|) of at least 10 mmHg and with early atherosclerotic markers in a subanalysis. We found that 6.4% of outpatients were left handed, and the percentage of individuals with systolic |ΔBP| (|ΔSBP|) and diastolic |ΔBP| (|ΔDBP|) of at least 10 mmHg was 14.4 and 7.2%, respectively. The dominant hand was not significantly associated with |ΔBP| of at least 10 mmHg or early atherosclerotic markers. This study suggests that BP measured in one arm is substitutable with that of the other arm because of a lack of association of |ΔBP| with the dominant hand. However, BP of both arms should be actively measured in new outpatients with moderate fever, lifestyle-related diseases, vascular events, age 65 years and above, and smoking history, all of which are factors potentially associated with |ΔBP| of at least 10 mmHg, regardless of the dominant hand.


Case mix use in 25 countries: a migration success but international comparisons failure.
Diagnosis related groups (DRGs) were the first health management tool to group patients in clinical meaningful categories with homogeneous resources consumption. They belong to case mix measures that have been introduced in the USA in the seventies to identify clinical differences in groups of patients as well as to document variation in hospitals and physicians practices by comparing similar groups with each others. They are also used to adjust payment rates and to improve quality of care. This innovative concept migrated from the USA to several Western European countries and to Australia first, to Eastern Europe and Asia soon thereafter. The main objective in most countries concerns a hospital financing new methodology, based on similar groups of patients, by applying a global budget. The main result of this international overview of 25 countries is to show the difficulty to make international comparisons, mainly because of the absence of a universal classification system for procedures. Furthermore, diagnostic classification systems to group patients vary also by country. These obstacles to compare hospital and physician practices have profound roots in cultural differences and in political sensitivity for organising health care delivery between countries. The development of an appropriate Case Mix grouper tool including uniform diagnoses and procedures codes, leading to international use becomes a public health priority.


Acupuncture treatment for functional gastrointestinal disorders: Identification of major acupoints using network analysis.
Using network analysis, we sought to determine the acupoints most commonly used to treat functional gastrointestinal disorders (FGIDs), particularly functional dyspepsia (FD) and irritable bowel syndrome (IBS). To explore the acupoint patterns used for FGID, data on acupoint combinations for FD and IBS were gathered from systematic reviews. Network analysis was used to determine the degree, closeness centrality, betweenness centrality, and eigenvector centrality of each acupoint. The most common acupoint combinations for FD and IBS were examined based on the eigenvector centrality. Network analysis revealed that CV12, ST25, ST36, CV10, and LR3, which had the highest eigenvector centrality values, were the main acupoints for treating FGID. CV12 was the main acupoint for treating FD, while ST25 was the hub acupoint for treating IBS in the abdomen. ST36, LR3, and PC6 were the key peripheral acupoints for FD and IBS. Using network analysis, we provided data that will aid the selection of both general and specific acupoints for FD and IBS, along with spatial information (i.e., the positions of acupoints on a body map). These findings could be applied in future acupuncture research on therapy for gastrointestinal system dysfunction. They may also help bridge the gap between the traditional meridian theory, which assumes that there is a link between diseases/symptoms and the specific body region being treated, and real-world clinical evidence.


Prescribing of disease modifying agents in older adults with multiple sclerosis.
The use of disease-modifying agents (DMAs) to treat Multiple Sclerosis (MS) in older adults is debated as the disease activity decreases with aging. However, limited data exist regarding prescribing patterns of DMAs among older adults with MS. To examine prescribing patterns of DMAs and the factors associated with DMA prescribing practices among older adults with MS using electronic medical records (EMR) data. A retrospective longitudinal cohort study was conducted using the TriNetX, a federated EMR network from the US, data from 2016 to 2019. The study included older adults (≥60 years) with MS diagnosis and at least one prescription record during the study period. Patients with DMA prescriptions were identified and further classified into injectable, oral, or infusion users based on their last DMA prescription. A multivariable logistic regression model was used to evaluate the factors associated with prescribing of DMAs. A multinomial logistic regression model was also used to determine the factors associated with prescribing a particular dosage form of DMA. The study cohort consisted of 12,922 older adults with MS, with 2,455 (18.99%) receiving DMA prescriptions. The commonly prescribed DMAs were injectables (10.46%), followed by orals (6.06%) and infusions (2.40%). Multivariable logistic regression revealed that older adults between 60- to 64 years (Adjusted Odds Ratio [aOR]= 2.38) and 65-69 years (aOR=1.60) had higher odds of receiving DMA compared to older adults of 70 years and above. African Americans (aOR=1.71) had higher odds of receiving DMA prescriptions compared to Caucasians. The presence of symptoms (pain, fatigue, speech, walking difficulty) and use of symptomatic medication (anti-fatigue medication, bladder dysfunction medication, antispasmodics, antidepressants, and relapse medication) increased the odds of being prescribed DMAs. Multinomial logistic regression found that patients 60-64 years of age had higher odds of being prescribed infusion (aOR, 95% Confidence Interval [CI] =2.06, 1.35-3.15) and oral (65-69 years: aOR=1.60, 1.24-2.07) over injectable DMAs compared to the older adults aged 70 years and above.Older males (aOR=1.68, 95% CI: 1.23-2.30) were associated with increased odds of being prescribed infusion DMA over injectable DMA compared to females. The presence of comorbidities such as coagulopathy and peripheral vascular disorders decreased the odds of being prescribed oral DMA over injectable DMA. Patients with cerebellar symptoms had an increased likelihood of being prescribed with an infusion DMA over injectable DMA. Patients using drugs for treating relapses had higher odds of being prescribed an infusion DMA over an injectable DMA. In terms of healthcare utilization, older adults with outpatient visits had higher odds of being prescribed an infusion DMA over an injectable DMA, while older adults with inpatient visits had lower odds of being prescribed an infusion DMA over an injectable DMA. Nearly one in five older adults with MS are prescribed DMAs, with a majority receiving injectable DMAs. Several demographic and clinical factors were associated with DMA prescribing . This study fills the data gap regarding the utilization of DMAs in older adults with MS.


Effect of Co-culture of mesenchymal stem cell and glomerulus endothelial cell to promote endothelialization under optimized perfusion flow rate in whole renal ECM scaffold.
In recent era, many researches on implantable bio-artificial organs has been increased owing to large gap between donors and receivers. Comprehensive organ based researches on perfusion culture for cell injury using different flow rate have not been conducted at the cellular level. The present study investigated the co-culture of rat glomerulus endothelial cell (rGEC) and rat bone marrow mesenchymal stem cells (rBMSC) to develop micro vascularization in the kidney scaffolds culturing by bioreactor system. To obtain kidney scaffold, extracted rat kidneys were decellularized by 1% sodium dodecyl sulfate (SDS), 1% triton X-100, and distilled water. Expanded rGECs were injected through decellularized kidney scaffold artery and cultured using bioreactor system. Vascular endothelial cells adhered and proliferated on the renal ECM scaffold in the bioreactor system for 3, 7 and 14 days. Static, 1 ml/min and 2 ml/min flow rates (FR) were tested and among them, 1 ml/min flow rate was selected based on cell viability, glomerulus character, inflammation/endothelialization proteins expression level. However, the flow injury was still existed on primary cell cultured at vessel in kidney scaffold. Therefore, co-culture of rGEC + rBMSC found suitable to possibly solve this problem and resulted increased cell proliferation and micro-vascularization in the glomerulus, reducing inflammation and cell death which induced by flow injury. The optimized perfusion rate under rGEC + rBMSC co-culture conditions resulted in enhanced endocellularization to make ECM derived implantable renal scaffold and might be useful as a way of treatment of the acute renal failure.


[Use of the rapid antigen technique in the diagnosis of Streptococcus pyogenes pharyngotonsillitis].
Streptococcus pyogenes is the most frequent bacterial cause of acute tonsillopharyngitis. The validity of the rapid antigen test was analysed for its diagnosis in a Paediatric Primary Care setting. The clinical profile with better diagnostic yield was also identified. The unnecessary use of antibiotics was quantified when the rapid antigen test or only the clinical diagnosis was used. The sensitivity of the assay to penicillin, erythromycin and clindamycin was also determined. Cross-sectional study was conducted on children between 2 to 14 years with acute tonsillitis and/or pharyngitis seen in five Primary Care Centres, from January 2008 to May 2010. After a clinical diagnosis, two swabs were taken for pharyngotonsillar smears: the first was used for a rapid antigen test, and the second one for a culture and a study of antibiotic sensitivity, with its analysis being blind to the rapid test result. A total sample of 546 consecutive was envisaged and with consecutive sampling. A total 192 patients were included. The prevalence of Streptococcus pyogenes was 38.7% (95% CI: 31.4-45.7). Odynophagia and scarlatiniform rash were most likely with positive cultures, the Streptococcus pyogenes was sensitive to penicillin in 100%, to erythromycin in 97.3% and to clindamycin in 86.3%. The specificity of the rapid antigen test was 91.5% and with a Negative Predictive Value of 91.5%. About half (49.2%) of those who would have receive antibiotics for clinical suspicion would have been treated unnecessarily, with this decreasing to at least in 29.5% when using the rapid antigen test. The rapid antigen test can lead to a better use of antibiotics. Its use in Paediatric Primary Health Care could be useful when, as when the result is negative, there would be no need to confirm by a culture, in those Health Centres with difficult access to laboratory.


Development of normal and dystrophic chick muscle in tissue culture. Production and release of creatine kinase and acetylcholinesterase.
Creatine kinase (CK) and acetylcholinesterase (AChE) were followed during the course of development of embryonic muscle in tissue culture for periods of up to 8 weeks. Control muscle cultures released CK and AChE into the medium for the first 2-3 weeks, after which time measurable enzyme release ended. In dystrophic muscle cultures release of CK and AChE continued over most of the 8-week culture period. Cumulative measurable activity of CK was significantly greater than that of controls by the 3rd week and thereafter and cumulative AChE release was greater than that of controls by the 5th week and averaged about twice that of controls by the end of the culture period. Total cell CK activity of the dystrophic muscle cultures was greater than control values but results of cellulose acetate electrophoretic analysis of CK isozyme composition indicated that control muscle cultures attained higher percentage levels of MM isozyme type and lower levels of MB isozyme. Breast muscle extracts from adult dystrophic chickens had an approximately 10--13% content of MB isozyme while in similar extracts of control chicken muscle only the MM isozyme was detectable and total activity was greater than in dystrophics. Bound AChE activity of dystrophic muscle was significantly greater than control levels subsequent to the 2nd week in culture. Soluble AChE activity of dystrophic muscle was somewhat greater than comparable control activity in 2 of 3 tissue culture series at approximately the same period. In 7--10 week-old dystrophic chickens, both soluble and bound AChE activities of dystrophic breast muscle extracts were markedly increased over control values.


The independent and combined effects of weight loss and aerobic exercise on blood pressure and oral glucose tolerance in older men.
The independent and combined effects of aerobic exercise training (AEX) and weight loss (WL) on blood pressure (BP) and oral glucose tolerance were studied in 97 nondiabetic, sedentary, older men. The men were assigned to either a WL (n = 31), AEX (n = 35), AEX+WL (n = 20) intervention, or to a control (CON) group (n = 11). The AEX regimen consisted of 3 days/week for 9 months, whereas WL consisted of a behavioral modification weight loss program that met weekly for 9 months. Maximal oxygen uptake increased significantly (P < .05) in both the AEX and AEX+WL groups (delta change: 0.45 +/- 0.06 and 0.43 +/- 0.07 L/min, respectively; mean +/- SEM), but did not change in the WL (delta change: 0.00 +/- 0.04 L/min) and CON (-0.05 +/- 0.07 L/min) groups. The AEX+WL and WL groups had comparable reductions in body weight (-8.6 +/- 0.7 and -8.8 +/- 0.7 kg,) and %fat (-5.4 +/- 0.5 and -5.2 +/- 0.6%) that were significantly greater than the CON and AEX groups, in which neither body weight nor %fat changed. The AEX, WL, and AEX+WL groups had significant reductions in both systolic (-6.4 +/- 1.3, -9.2 +/- 1.6, and -9.7 +/- 2.2 mm Hg, respectively) and diastolic (-5.2 +/- 1.0, -6.2 +/- 1.3, and -7.2 +/- 1.5 mm Hg, respectively) BP after their respective interventions compared with the change in systolic (3.2 +/- 3.6 mm Hg) and diastolic (-0.5 +/- 1.9 mm Hg) BP in the CON group. The AEX, WL, and AEX+WL groups also had significant reductions in glucose and insulin responses to an oral glucose tolerance test compared with the CON group. The reduction in the insulin area in the AEX+WL group was significantly greater than in the AEX and WL groups. The results of this study demonstrate that AEX and WL are effective nonpharmacologic therapies to lower BP and alter glucose and insulin responses to an oral glucose challenge. However, combining the two interventions does not result in any further improvement in BP.


Transcriptome dynamics of a broad host-range cyanophage and its hosts.
Cyanobacteria are highly abundant in the oceans and are constantly exposed to lytic viruses. The T4-like cyanomyoviruses are abundant in the marine environment and have broad host-ranges relative to other cyanophages. It is currently unknown whether broad host-range phages specifically tailor their infection program for each host, or employ the same program irrespective of the host infected. Also unknown is how different hosts respond to infection by the same phage. Here we used microarray and RNA-seq analyses to investigate the interaction between the Syn9 T4-like cyanophage and three phylogenetically, ecologically and genomically distinct marine Synechococcus strains: WH7803, WH8102 and WH8109. Strikingly, Syn9 led a nearly identical infection and transcriptional program in all three hosts. Different to previous assumptions for T4-like cyanophages, three temporally regulated gene expression classes were observed. Furthermore, a novel regulatory element controlled early-gene transcription, and host-like promoters drove middle gene transcription, different to the regulatory paradigm for T4. Similar results were found for the P-TIM40 phage during infection of Prochlorococcus NATL2A. Moreover, genomic and metagenomic analyses indicate that these regulatory elements are abundant and conserved among T4-like cyanophages. In contrast to the near-identical transcriptional program employed by Syn9, host responses to infection involved host-specific genes primarily located in hypervariable genomic islands, substantiating islands as a major axis of phage-cyanobacteria interactions. Our findings suggest that the ability of broad host-range phages to infect multiple hosts is more likely dependent on the effectiveness of host defense strategies than on differential tailoring of the infection process by the phage.


The process of definitive institutionalization of community dwelling demented vs non demented elderly: data obtained from a network of sentinel general practitioners.
The aim of the present study was to investigate the differences between characteristics of community dwelling demented and non demented elderly and their caregivers at the moment of definitive institutionalization. The study is a cross-sectional analysis performed on data obtained from a Network of Sentinel General Practitioners. Older demented patients with several concomitant diseases were mainly placed because of unmanageable behavioral disturbances. Strikingly, these dementia patients were more often confronted with a time delay in definitive institutionalization due to their high care dependence. Although burden in the dementia family caregivers was an important motivation for definitive institutionalization, it did not seem to be a motive in the final institutionalization circumstances. Behavioral disturbances are independently of caregiver burden, professional support, or a spousal relationship the main direct reason for institutionalization of community dwelling demented elderly. The way caregivers feel supported might play the key role in the final placement decision.


Immunologic, morphologic and chromosomal characterization of a cell line (TC78) established from a child with acute lymphoblastic leukemia.
We characterized a cell line established from bone marrow cells from a child with acute lymphoblastic leukemia. This cell line, TC78, had lymphoblastic morphology and was cytoplasmic peroxidase and esterase negative. The cells did not have T- or B-cell properties such as E- or EAC-rosette forming ability, reactivity with monoclonal T-cell or B2 antibodies, or immunoglobulin synthesis. We concluded that TC78 was a pre-pre B-cell line based on the following monoclonal antibody staining pattern: BA-1+, BA-2+, cALLa+, Ia+, 2H7+ and OKB2+. Growth in 'Dickie' culture and reactivity with 1G10 myeloid antibody suggested coexpression of lymphoid and myeloid characteristics. However, 1G10 expression proved dependent on culture conditions, illustrating one caveat in application of monoclonal antibodies in lineage determination.


Neurochemical investigations of dopamine neuronal systems in iron-regulatory protein 2 (IRP-2) knockout mice.
Abnormal iron accumulations are frequently observed in the brains of patients with Parkinson's disease and in normal aging. Iron metabolism is regulated in the CNS by iron regulatory proteins (IRP-1 and IRP-2). Mice engineered to lack IRP-2 develop abnormal motoric behaviors including tremors at rest, abnormal gait, and bradykinesia at middle to late age (18 to 24 months). To further characterize the dopamine (DA) systems of IRP-2 -/- mice, we harvested CNS tissue from age-matched wild type and IRP-2 -/- (16-19 months) and analyzed the protein levels of tyrosine hydroxylase (TH), dopamine transporter (DAT), vesicular monoamine transporter (VMAT2), and DA levels in dorsal striatum, ventral striatum (including the core and shell of nucleus accumbens), and midbrain. We further analyzed the phosphorylation of TH in striatum at serine 40, serine 31, and serine 19. In both dorsal and ventral striatum of IRP-2 knockout mice, there was a 20-25% loss of TH protein and accompanied by a approximately 50% increase in serine 40 phosphorylation above wild-type levels. No change in serine 31 phosphorylation was observed. In the ventral striatum, there was also a significant loss (approximately 40%) of DAT and VMAT2. Levels of DA were decreased (approximately 20%) in dorsal striatum, but turnover of DA was also elevated ( approximately 30%) in dorsal striatum of IRP-2 -/- mice. We conclude that iron misregulation associated with the loss of IRP-2 protein affects DA regulation in the striatum. However, the modest loss of DA and DA-regulating proteins does not reflect the pathology of PD or animal models of PD. Instead, these observations support that the IRP-2 -/- genotype may enable neurobiological events associated with aging.


Crystal structure of catena-poly[[[trans-bis(aceto-nitrile-κN)diaquacobalt(II)]-μ-pyrazine-κ(2) N:N'] dinitrate].
The central structural motif of the title coordination polymer, [Co(NO3)2(C4H4N2)(CH3CN)2(H2O)2] n , is a chain composed of Co(II) ions linked by bis-monodentate bridging pyrazine ligands through their N atoms. The Co(II) ion is located on an inversion center and is additionally coordinated by two O atoms of water mol-ecules and two N atoms of aceto-nitrile mol-ecules. The resultant N4O2 coordination sphere is distorted octa-hedral. The linear cationic chains extend parallel to the a axis and are aligned into layers parallel to the ac plane. Nitrate anions are situated in the space between the Co(II) chains and form O-H⋯O hydrogen bonds with the coordinating water mol-ecules, leading to a three-dimensional network structure. Weak C-H⋯O hydrogen bonds are also present between pyrazine or aceto-nitrile mol-ecules and the nitrate anions.


Magnetic field detection limits for ultraclean graphene Hall sensors.
Solid-state magnetic field sensors are important for applications in commercial electronics and fundamental materials research. Most magnetic field sensors function in a limited range of temperature and magnetic field, but Hall sensors in principle operate over a broad range of these conditions. Here, we evaluate ultraclean graphene as a material platform for high-performance Hall sensors. We fabricate micrometer-scale devices from graphene encapsulated with hexagonal boron nitride and few-layer graphite. We optimize the magnetic field detection limit under different conditions. At 1 kHz for a 1 μm device, we estimate a detection limit of 700 nT Hz-1/2 at room temperature, 80 nT Hz-1/2 at 4.2 K, and 3 μT Hz-1/2 in 3 T background field at 4.2 K. Our devices perform similarly to the best Hall sensors reported in the literature at room temperature, outperform other Hall sensors at 4.2 K, and demonstrate high performance in a few-Tesla magnetic field at which the sensors exhibit the quantum Hall effect.


3D-QSAR analysis of antimalarial farnesyltransferase inhibitors based on a 2,5-diaminobenzophenone scaffold.
With annual death tolls in the millions and emerging resistance to existing drugs, novel therapies are needed against malaria. Wiesner et al. recently developed a novel class of antimalarials derived from farnesyltransferase inhibitors based on a 2,5-diaminobenzophenone scaffold. The compounds displayed a wide range of activity, including submicromolar, against the multi-drug resistant Plasmodium falciparum strain Dd2. In order to investigate quantitatively the local physicochemical properties involved in the interaction between drug and biotarget, we used the 3D-QSAR methods CoMFA and CoMSIA to study some of the series, including the screened lead compound 2,5-bis-acylaminobenzophenone, 28 cinnamic acid derivatives, 29 N-(3-benzoyl-4-tolylacetylaminophenyl)-3-(5-aryl-2-furyl)acrylic acid amides, and 34 N-(4-substituted-amino-3-benzoylphenyl)-[5-(4-nitrophenyl)-2-furyl]acrylic acid amides. We found that steric, electrostatic, and hydrophobic properties of substituent groups play key roles in the bioactivity of the series of compounds, while hydrogen bonding interactions show no obvious impact. We built several highly predictive 3D-QSAR models, including a CoMSIA one composed of steric, electrostatic, and hydrophobic fields, with r(2)=0.94, q(2)=0.63, and r(pred)(2)=0.63. The results provide insight for optimization of this class of antimalarials for better activity and may prove helpful for further lead optimization.


Raga todi intervention on state anxiety level in female young adults during COVID-19.
Raga todi, an Indian classical music raga is emerging as a therapeutic approach in mental health and well being. This study aims to assess the relationship of raga todi and state anxiety at the time of novel coronavirus spread across the globe. 30 young female adults of age 20-28 years were randomly assigned to experimental group and control group of 15 participants each to which a pre and post test of State Anxiety test of STAI-A was administered. Results stated that state anxiety is negatively correlated to the listening of raga todi. Thus, listening to raga todi could be useful in reducing the state anxiety level provoked by a stressful life event like the lethal coronavirus pandemic.


Design and synthesis of 14 and 15-membered macrocyclic scaffolds exhibiting inhibitory activities of hypoxia-inducible factor 1α.
Inspired by the privileged molecular skeletons of 14- and 15-membered antibiotics, we adopted a relatively unexplored synthetic approach that exploits alkaloidal macrocyclic scaffolds to generate modulators of protein-protein interactions (PPIs). As mimetics of hot-spot residues in the α-helices responsible for the transcriptional regulation, three hydrophobic sidechains were displayed on each of the four distinct macrocyclic scaffolds generating diversity of their spatial arrangements. Modular assembly of the building blocks followed by ring-closing olefin metathesis reaction and subsequent hydrogenation allowed concise and divergent synthesis of scaffolds 1-4. The 14-membered alkaloidal macrocycles 2-4 demonstrated similar inhibition of hypoxia-inducible factor (HIF)-1α transcriptional activities (IC50 between 8.7 and 10 µM), and 4 demonstrated the most potent inhibition of cell proliferation in vitro (IC50 = 12 µM against HTC116 colon cancer cell line). A docking model suggested that 4 could mimic the LLxxL motif in HIF-1α, in which the three sidechains are capable of matching the spatial arrangements of the protein hot-spot residues. Unlike most of the stapled peptides, the 14-membered alkaloidal scaffold has a similar size to the α-helix backbone and does not require additional atoms to induce α-helix mimetic structure. These experimental results underscore the potential of alkaloidal macrocyclic scaffolds featuring flexibly customizable skeletal, stereochemical, substitutional, and conformational properties for the development of non-peptidyl PPI modulators targeting α-helix-forming consensus sequences responsible for the transcriptional regulation.


Lateral meniscus allograft transplantation: an arthroscopically-assisted single-incision technique using all-inside sutures with a suture hook.
We present an arthroscopically-assisted single-incision technique using all-inside sutures with a suture hook in the lateral meniscus allograft transplantation. Although this technique is technically demanding, it provides vertically oriented and secure sutures with good tissue approximation without the accessory skin incision.


Expression analyses of Arabidopsis oligopeptide transporters during seed germination, vegetative growth and reproduction.
AtOPT promoter-GUS fusions were constructed for six of the nine known, putative oligopeptide transporters (OPTs) in Arabidopsis thaliana and used to examine AtOPT expression at various stages of plant development. AtOPT1, AtOPT3, AtOPT4, AtOPT6 and AtOPT7 were expressed in the embryonic cotyledons prior to root radicle emergence. Except for AtOPT8, which gave weak expression, all AtOPTs were strongly expressed in post-germinative seedlings with strongest expression in vascular tissues of cotyledons and hypocotyls. Preferential expression of AtOPTs in vascular tissues was also observed in cotyledons, leaves, hypocotyls, roots, flowers, siliques, and seed funiculi of seedlings and adult plants. Differential tissue-specific expression was observed for specific AtOPTs. For example, AtOPT1, AtOPT3 and AtOPT8 were uniquely expressed in pollen. Only AtOPT1 was expressed in growing pollen tubes, while only AtOPT6 was observed in ovules. AtOPT8 was transiently expressed in seeds during early stages of embryogenesis. Iron limitation was found to enhance expression of AtOPT3. These data suggest distinct cellular roles for specific AtOPTs including nitrogen mobilization during germination and senescence, pollen tube growth, pollen and ovule development, seed formation and metal transport.


Direct-injection LC-MS-MS method for high-throughput simultaneous quantitation of simvastatin and simvastatin acid in human plasma.
A direct-injection liquid chromatography-mass spectrometry-mass spectrometry (LC-MS-MS) method was developed and validated for the simultaneous quantitation in human plasma of the widely used cholesterol-lowering prodrug simvastatin and its in vivo generated active drug, simvastatin acid. The plasma samples were injected into the LC-MS-MS system after simply adding the internal standard solution in an aqueous buffer and centrifuging. The analytes in the buffered plasma samples were found to be stable for at least 24 h at 4 degrees C. The method was successfully validated under the challenging condition of using a large number of quality control (QC) samples including those in which the ratio of the simvastatin concentration to the simvastatin acid concentration was different from the concentration ratio in the calibration curve standards. Under the dual stabilizing conditions of lower temperature (4 degrees C) and lower plasma pH of 4.9, the in-process hydrolysis of simvastatin to simvastatin acid or the lactonization of simvastatin acid to simvastatin was minimized to < or = 1.0%. Although the entire run time for on-line cleanup and analysis was only 2.5 min, chromatographic base-line separation of simvastatin from simvastatin acid, which was required to avoid the interference by simvastatin acid with the simvastatin selected reaction monitoring channel, was achieved. The desired lower limit of quantitation of 0.5 ng/ml was achieved by injecting only an equivalent of 8.0 microl of the plasma sample. The extraction column lasted for at least 500 injections.


Timing between ejaculations changes paternity success.
It is believed that when two male rats copulate with a single female, the last one that ejaculates fathers significantly more pups than the first one. To determine the effects of order and elapsed time between two males copulating with the same female, albino Wistar (W) and pigmented Long-Evans (LE) male rats copulated with a W female. Paternity was confirmed by the pups' pigmentation. Three groups were made according to the elapsed time between the first male's ejaculation and the placement of the female with the second male; T0 (0-min group); T5 (5-min group); and T10 (10-min group). Male order was counterbalanced in each group. The results showed that the second male had the advantage in T0, but the first males had the advantage in T5 and T10. These data indicate that in a two-male situation, the advantage in paternity for a second male occurs only during copulations following the first few minutes of a first male's ejaculation. After 5 min, a second male has no advantage in fathering pups.


Antifungal efficacy of soft contact lens disinfecting solutions against Fusarium solani and Candida albicans.
The aim was to evaluate the disinfection properties of six multi-purpose contact lens disinfection solutions (MPDS) available in Iran against Fusarium solani and Candida albicans, based on the International Organization for Standardization (ISO) 14729 guidelines. Three lots each of six multi-purpose solutions, namely, Renu MultiPlus, Solo Care Aqua, All-Clean soft, Contact All In One Advanced, Hippia, Ginza Multi-Purpose Solution and 0.9% normal saline solution were inoculated with standard strains of Fusarium solani (ATCC 36031) and Candida albicans (ATCC 10231). Surviving fungi were determined by counting colony-forming units on recovery plates at specified times. The mean log reduction in fungal numbers at the manufacturer's minimum recommended disinfection time was determined and compared with the criteria for stand-alone disinfection products for each MPDS against each fungal strain. Renu MultiPlus, Contact All In One Advanced and All-Clean Soft were effective in achieving the mean 1.0 log reduction in fungal numbers, based on ISO 14729 stand-alone primary acceptance criteria. Solo Care Aqua failed to achieve the ISO 14729 stand-alone primary acceptance criteria for Candida albicans. The other solutions were not effective against test microorganisms after the specified times. There were differences in the rates of disinfection efficacy. Generally, Renu MultiPlus, Contact All In One Advanced and All-Clean Soft exceeded the primary acceptance criteria of the ISO 14729 guidelines for stand-alone contact lens solutions.


Article Topic: Positioning Ulcerative Colitis Therapies in 2022 and Beyond.
Ulcerative colitis (UC) is a chronic disease with an increasing incidence. Recent guidelines emphasize treating toward objective targets, requiring the use of effective, steroid-sparing therapies. This review summarizes the safety and efficacy data of available therapies as well comparative effectiveness studies in order to help the reader make rational treatment decisions. Following the approval of tumor necrosis factor alpha antagonists, we have seen recent regulatory approval of several additional biologic and small molecule agents from several therapeutic classes (integrin antagonists, interleukin 12/23 antagonists, Janus kinase inhibitors, and sphingosine-1-phosphate receptor antagonists) for UC. Randomized, controlled trials, real-world analyses, and network meta-analyses have investigated the comparative safety and efficacy of these therapies in order to help clinicians better position these therapies in clinical practice. Numerous agents are now approved for the treatment of UC. This evidence-based review will help the reader understand the important factors weighing into treatment decisions for patients with UC and enable patient education and discussion with a focus on a shared decision-making approach.


Cytogenetic effects of cyclamates.
PHA-stimulated human peripheral lymphocytes were used as a model system for assessing the in vitro effects of calcium cyclamate. Techniques of autoradiography, cytological staining, cell counting, liquid scintillation and karyotyping were used to study the cytogenetic damage and biochemical effects of calcium cyclamate when assayed in 24 hour intervals for 96 hours. The cells were exposed to 10(-2) and 10(-3) molar concentrations of calcium cyclamate in TC 199 medium with fetal calf serum and antibiotics. These studies were carried out in three (3) phases. Phase I was primarily orientation studies of the effects of cyclamates and included running preliminary test checks, the establishment of parameters of dosage, assessing growth patterns and selecting key chromosomal aberrations. Sixty four (64) of the metaphase spreads showed morphologically detectable changes and aberrations. It was also noted that the addition of cyclamate increased mitotic rate of lymphocyte cells in cultures. Phase III arranged research designs to determine more precise characterization of chromosomal observations and morphological effects. Among other findings it was noted that of 13 types of observations only ten were found in the experimental group. The introduction of cyclamates increased the stability of the leucocyte cultures. These studies reinforced the findings on the increase of mitotic rate. Phase III extended protocols to include autoradiography and scintillation counting. It was determined that calcium cyclamate impaired the synthesis of deoxribonunucleic acid (as depicted by decreased incorporation of tritiated thymidine), reduced grain counts in autoradiographs and increased chromosome aberrations in cyclamate treated PHA stimulated peripheral blood lymphocytes in vitro. Morphological changes and growth rates showed significant effects. These studies indicate that calcium cyclamate has variable significant effects on leucocytes growth and chromosome morphology.


Inflammation triggers specific microRNA profiles in human adipocytes and macrophages and in their supernatants.
The relevance of microRNAs (miRNAs) in adipose tissue is increasingly recognized, being intrinsically linked to different pathways, including obesity-related inflammation. In this study, we aimed to characterize the changes induced by inflammation on the miRNA pattern of human adipocytes and macrophages. Therefore, an extensive profile of 754 common miRNAs was assessed in cells (human primary mature adipocytes, and the macrophage-like cell line THP-1) and in their supernatants (SN) using TaqMan low-density arrays. These profiles were evaluated at the baseline and after administration of lipopolysaccharide (LPS, 10 ng/ml) and LPS-conditioned medium from M1 macrophages (MCM, 5%). The miRNAs that experienced the most dramatic changes were studied in subcutaneous human adipose tissue before and approximately 2 years after bariatric surgery-induced weight loss. Differentiated adipocytes expressed 169 miRNAs, being 85 detectable in the SN. In M1 macrophages, 183 miRNAs were detected, being 106 also present in the SN. Inflammation led to an increased number of miRNAs detectable in cells and in their SNs in both adipocytes (+8.3% and +24.7%) and M1 macrophages (+1.4% and +5%, respectively). Indeed, under inflammatory conditions, adipocytes and M1 macrophages shared the expression of 147 (+9%) miRNAs, and 100 (+41%) common miRNAs were found in their SNs. Twelve of these factors were also linked to inflammation in whole adipose tissue from obese subjects. Interestingly, miR-221 (2-fold, P = 0.002), miR-222 (2.5-fold, P = 0.04), and miR-155 (5-fold, P = 0.015) were increased in inflamed adipocytes and in their SNs (15-, 6-, and 4-fold, respectively, all P < 0.001). Furthermore, their expressions in human adipose tissue concordantly decreased after weight loss (-51%, P = 0.003, -49%, P = 0.03, and -54.4%, P = 0.005, respectively). Inflammation induces a specific miRNA pattern in adipocytes and M1 macrophages, with impact on the physiopathology of obesity-induced inflammation of adipose tissue. The crosstalk between cells should be investigated further.


Epiploic gonadal vein as a new bypass route for extrahepatic portal venous obstruction: report of a case.
A 61-year-old man was referred to our hospital to treat extrahepatic portal venous obstruction. Endoscopic injection sclerotherapy (EIS) was performed for the esophageal varices; however, the patient returned with massive hematemesis from gastric varices 6 months after treatment. Although the varices were treated with EIS, gastric devascularization and splenectomy concomitant with shunt surgery were required to treat uncontrollable, frequent diarrhea and abdominal distension. Because the splenic vein, left gastric vein, left portal vein, and inferior vena cava were inadequate for anastomosis, an epiploic gonadal vein bypass was performed. The bypass graft remains patent 7 months after surgery, and the patient is in good health without any clinical symptoms. We describe a new bypass route for extrahepatic portal venous obstruction.


Masked Amino Trimethyl Lock (H2 N-TML) Systems: New Molecular Entities for the Development of Turn-On Fluorophores and Their Application in Hydrogen Sulfide (H2 S) Imaging in Human Cells.
Masked trimethyl lock (TML) systems as molecular moieties enabling the bioresponsive release of compounds or dyes in a controlled temporal and spatial manner have been widely applied for the development of drug conjugates, prodrugs or molecular imaging tools. Herein, we report the development of a novel amino trimethyl lock (H2 N-TML) system as an auto-immolative molecular entity for the release of fluorophores. We designed Cou-TML-N3 and MURh-TML-N3 , two azide-masked turn-on fluorophores. The latter was demonstrated to selectively release fluorescent MURh in the presence of physiological concentrations of the redox-signaling molecule H2 S in vitro and was successfully applied to image H2 S in human cells.


[Clinical study and hormonal evaluation of a 48, XXYY male. Comparison with previously published cases].
A new case of the infrequent 48, XXYY syndrome is presented. The somatomedin C and HGH values were high, perhaps playing a role in the above of the mean height of Klinefeter patients, of which this cromosomic abnormality is a variant.


Explantation of phakic intraocular lenses: causes and outcomes.
To describe reasons for explantation of anterior and posterior chamber phakic intraocular lenses (pIOLs), as well as outcomes of the surgery. The medical files of patients who underwent pIOL explantation due to complications were reviewed. All patients were divided into three groups based on the type of explanted pIOL: anterior chamber angle-supported (AS pIOL), anterior chamber iris-fixated (IF pIOL), and posterior chamber (PC pIOL). Sixty-two eyes of 41 patients were evaluated. There were 26 (41.9%), 16 (25.8%), and 20 (32.2%) eyes in the AS pIOL, IF pIOL, and PC pIOL groups, respectively. The mean interval between the implantation and removal of the pIOL (i.e., survival time) was 13.6 ± 8.1 years (range 0.01-21.22 years). The mean follow-up after the explantation was 22.5 ± 4.0 months (range 11.3-28.7 months). The main causes of explantation were cataract in the PC pIOL group (60%) and chronic endothelial cell loss in the AS pIOL group (53.8%) and IF pIOL group (56.2%). Overall, the procedure most often combined with pIOL explantation was phacoemulsification and implantation of a posterior chamber IOL (40.3%), followed by keratoplasty (9.6%). Intraoperative complications were significantly more common in the AS pIOL group than the other groups (p < 0.001). Explantation of anterior chamber pIOLs due to severe endothelial cell loss and the proportion of keratoplasty was more common in patients with a relatively long survival time. Therefore, patients with pIOL implantation should be monitored regularly after surgery.


Serum lipid and lipoprotein abnormalities in human African trypanosomiasis.
We have studied the serum lipoprotein system in human African trypanosomiasis (Trypanosoma brucei gambiense infection). The study was carried out on 74 Congolense patients suffering from sleeping sickness and 34 Congolense control subjects living in the endemic region of Boko Songho. We have determined the serum concentrations of lipids (triglycerides, cholesterol, phospholipids) and apolipoproteins (apolipoprotein A-I and B), and the separation of serum lipoproteins by electrophoresis. For the patients infected with T. b. gambiense, in comparison with control subjects, the results have shown (i) a significant increase in triglyceride concentration and a decrease in cholesterol concentration; (ii) a significant rise in apolipoprotein B concentration and a significant reduction in apolipoprotein A-I concentration; and (iii) an increase in low density lipoproteins and a decrease in high density lipoproteins. We conclude, therefore, that human African trypanosomiasis is associated with marked alterations in the composition and levels of host lipoproteins.


Influence of peripherally-administered peptides on ethanol-induced gastric ulcers in the rat.
1. Subcutaneous administration of bombesin (50-200 micrograms/kg), cholecystokinin-8 (10-100 micrograms/kg) and epidermal growth factor (10-100 micrograms/kg) but not calcitonin gene-related peptide (25 micrograms/kg), neurotensin (500 micrograms/kg) or substance P (200 micrograms/kg) produced a marked and dose-related inhibition of ethanol-induced gastric lesions in rats. 2. These actions were inhibited by a non-ulcerogenic dose of indomethacin suggesting that prostaglandins are involved in the protective activity of bombesin, cholecystokinin-8 and epidermal growth factor.


[Bilateral failure of the vestibular system of the inner ear underlying an equilibrium disturbance].
Bilateral vestibular failure is a fairly rare, though possible cause underlying the symptoms of a dizzy patient. A recognizable oscillopsia symptom makes the diagnosis easier, though confirmation of the diagnosis requires a more precise measurement. The prognosis for recovery is not good, and curative treatment is not available. Diagnosis is nevertheless important with regard to rehabilitation and maintaining of functional capacity.


Magnetic resonance enterography for predicting the clinical course of Crohn's disease strictures.
Strictures are the most common Crohn's disease complication, but their natural history is unknown. This study aimed to characterize inflammation, predict prognosis, and understand the impact of drug therapy using magnetic resonance enterography (MRE). Patients with a stricture diagnosed on MRE over a 5-year period were reviewed for MRE disease extent and inflammation, clinical course, C-reactive protein, response to anti-TNF therapy, endoscopic dilatation, hospitalization, and surgery. 136 patients had 235 strictures (77, one and 59, ≥ 2 strictures). 46% of patients underwent surgery after a median 6 months; median follow-up for those not requiring surgery was 41 months. Predictors of surgery: Hospitalization because of obstruction predicted subsequent surgery (OR 2.50; 95% CI 1.06-5.90) while anti-TNF therapy commenced at stricture diagnosis was associated with a reduced risk (OR 0.23; 95% CI 0.05-0.99). MRE characteristics associated with surgery were proximal bowel dilatation ≥ 30-mm diameter (OR 2.98; 95% CI 1.36-6.55), stricture bowel wall thickness ≥ 10-mm (OR 2.42; 95% CI 1.11-5.27), and stricture length > 5-cm (OR 2.56; 95% CI 1.21-5.43). 81% of patients with these three adverse MRE features required surgery versus 17% if none were present (P < 0.001). Accuracy for these three MRE variables predicting surgery was high (AUC 0.76). Magnetic resonance enterography findings in Crohn's disease strictures are highly predictive of the disease course and the need for future surgery. MRE may also identify who would benefit from treatment intensification. Anti-TNF therapy is associated with reduced risk of surgery and appears to alter the natural history of this complication.


Optimising Intraoperative Fluid Management in Patients Treated with Adolescent Idiopathic Scoliosis-A Novel Strategy for Improving Outcomes.
Scoliosis surgery is a challenge for the entire team in terms of safety, and its accomplishment requires the utilization of advanced monitoring technologies. A prospective, single centre, non-randomised controlled cohort study, was designed to assess the efficacy of protocolised intraoperative haemodynamic monitoring and goal-directed therapy in relation to patient outcomes following posterior fusion surgery for adolescent idiopathic scoliosis (AIS). The control group (n = 35, mean age: 15 years) received standard blood pressure management during the surgical procedure, whereas the intervention group (n = 35, mean age: 14 years) underwent minimally invasive haemodynamic monitoring. Arterial pulse contour analysis (APCO) devices were employed, along with goal-directed therapy protocol centered on achieving target mean arterial pressure and stroke volume. This was facilitated through the application of crystalloid boluses, ephedrine, and noradrenaline. The intervention group was subjected to a comprehensive protocol following Enhanced Recovery After Surgery (ERAS) principles. Remarkably, the intervention group exhibited notable advantages (p < 0.05), including reduced hospital stay durations (median 7 days vs. 10), shorter episodes of hypotension (mean arterial pressure < 60 mmHg-median 8 vs. 40 min), lesser declines in postoperative haemoglobin levels (-2.36 g/dl vs. -3.83 g/dl), and quicker extubation times. These compelling findings strongly imply that the integration of targeted interventions during the intraoperative care of AIS patients undergoing posterior fusion enhance a set of treatment outcomes.


Association between family environment and attention deficit hyperactivity disorder in children--mothers' and teachers' views.
To ascertain whether factors of the family environment and gestational period are associated with the appearance of ADHD in children, as reported by various different informants (mothers and teachers). This paper presents results from the dataset of a longitudinal study to evaluate behavioral problems among schoolchildren in São Gonçalo, Rio de Janeiro State, in 2005 and 2006. The cross-section considered for this paper comprises records of exposure factors and ADHD. In all, 370 schoolchildren of the public school system were assessed by 3-stage cluster sampling. The Child Behavior Checklist (CBCL) and the Teacher Report Form (TRF) were used to measure outcomes. The exposure factors examined were: profile of child and mother, variables relating to the family environment, and perinatal considerations. The questions were answered by mothers and teachers. A hierarchical logistic regression model was used. Precariously functioning families, lack of social support for mothers, adverse life events and discord during pregnancy were the factors associated with mother-reported ADHD. When ADHD was reported by teachers, the variables selected were: Intelligence quotient (IQ) and sex, with children with low IQ scores and boys more likely to display the disorder. Assessment of ADHD by teachers or mothers reveals specific characteristics that reflect how each of these informants understands the children. This highlights the importance of using informants from different environments in diagnosing the disorder.


Association between in vivo bone formation and ex vivo migratory capacity of human bone marrow stromal cells.
There is a clinical need for developing systemic transplantation protocols for use of human skeletal stem cells (also known bone marrow stromal stem cells) (hBMSC) in tissue regeneration. In systemic transplantation studies, only a limited number of hBMSC home to injured tissues suggesting that only a subpopulation of hBMSC possesses "homing" capacity. Thus, we tested the hypothesis that a subpopulation of hBMSC defined by ability to form heterotopic bone in vivo, is capable of homing to injured bone. We tested ex vivo and in vivo homing capacity of a number of clonal cell populations derived from telomerized hBMSC (hBMSC-TERT) with variable ability to form heterotopic bone when implanted subcutaneously in immune deficient mice. In vitro transwell migration assay was used and the in vivo homing ability of transplanted hBMSC to bone fractures in mice was visualized by bioluminescence imaging (BLI). In order to identify the molecular phenotype associated with enhanced migration, we carried out comparative DNA microarray analysis of gene expression of hBMSC-derived high bone forming (HBF) clones versus low bone forming (LBF) clones. HBF clones were exhibited higher ex vivo transwell migration and following intravenous injection, better in vivo homing ability to bone fracture when compared to LBF clones. Comparative microarray analysis of HBF versus LBF clones identified enrichment of gene categories of chemo-attraction, adhesion and migration associated genes. Among these, platelet-derived growth factor receptor (PDGFR) α and β were highly expressed in HBF clones. Follow up studies showed that the chemoattractant effects of PDGF in vitro was more enhanced in HBF compared to LBF clones and this effect was reduced in presence of a PDGFRβ-specific inhibitor: SU-16 f. Also, PDGF exerted greater chemoattractant effect on PDGFRβ(+) cells sorted from LBF clones compared to PDGFRβ(-) cells. Our data demonstrate phenotypic and molecular association between in vivo bone forming ability and migratory capacity of hBMSC. PDGFRβ can be used as a potential marker for the prospective selection of hBMSC populations with high migration and bone formation capacities suitable for clinical trials for enhancing bone regeneration.


Squamous-cell carcinoma of the colon. Experience at the University of Chicago, review of the literature, report of two cases.
Primary squamous-cell and adenosquamous-cell carcinoma of the colon are uncommon and their characteristics not well known. This paper reports the clinical features and pathologic findings of two colonic adenosquamous carcinomas and reviews other reports of adenosquamous and squamous carcinoma of the colon from the English medical literature. Including these two cases, 63 cases have been reported since 1927. Of these, six occurred in patients with ulcerative colitis, three occurred at the colonic opening of chronic colocutaneous fistulas, and concomitant schistosomiasis was present in two patients. Synchronous squamous-cell carcinoma of the colon was present in 3.2 percent of cases and 10 percent had either antecedent, synchronous, or metachronous adenocarcinoma of the colon. These lesions appeared to be distributed uniformly throughout the colon. The five-year survival after resective therapy for primary squamous-cell and adenosquamous-cell carcinoma of the colon calculated with life table analysis is 50 percent for Dukes' B lesions, 33 percent for Dukes' C lesions, and 0 percent for Dukes' D lesions.


Bioactive triterpenoid saponins from the tubers of Hemsleya amabilis Diels.
Five new cucurbitane-type triterpenoid saponins Xuedanosides A-E (1-5) were isolated from the medicinal plant Hemsleya amabilis Diels by silica gel column, octadecylsilyl (ODS) column, and pre-HPLC techniques. Their structures were determined by spectroscopic analysis and examined alongside existing data from prior studies. Separated compounds were evaluated for their cytotoxic activity in HeLa, HCT-8, MCF-7 and HepG2 human cancer cell lines, and compounds 1 and 2 showed significant effects against HeLa cells with IC50 values of 3.21 and 8.57 μM, respectively.


Increased antibody-dependent cytotoxicity mediated by purified monocytes in Hodgkin's disease.
Monocyte antibody-dependent cytotoxicity was studied in 19 patients with Hodgkin's disease and 14 normal controls. This function was investigated after isolation of the monocytes by means of a modified elutriation technique. Direct sizing and counting of the cells present in the effluent enabled individual adjustment during each separation procedure. The absolute monocyte count in the peripheral blood of patients with Hodgkin's disease was higher (P less than 0.002) than in normal controls. Nearly 90% pure monocyte suspensions, representing 82% of all elutriated monocytes, were obtained. The elutriation characteristics of the monocytes in both groups were essentially the same, irrespective of marked interindividual differences. Kill of antibody-coated chicken red blood cells was measured by DNA flow cytometry. In comparison to normal controls, a significantly increased (P less than 0.0004), stage-independent, monocyte antibody-dependent cytotoxicity was found in patients with Hodgkin's disease. The percentage of kill in symptomatic patients tended to be higher than in the asymptomatic group; no correlation was found with the absolute number of circulating monocytes.


Site-specific recombination in Zea mays.
The elimination of marker genes after selection is recommended for the commercial use of genetically modified plants. We compared the applicability of the two site-specific recombination systems Cre/lox and Flp/FRT for marker gene elimination in maize plants. The selection marker gene pat surrounded by two identically directed lox or FRT sites was introduced into maize. Sexual crossing with plants harboring the corresponding constitutively expressed recombinase led to the precise and complete excision of the lox-flanked marker gene in the F1 progeny, whereas Flp-mediated recombination of FRT sequences occurred rarely. Further examination of site-specific integration was done by biolistic bombardment of immature embryos harboring only one lox site with a lox.uidA sequence with results indicating directed integration.


Nanosecond time-resolved fluorescence kinetic studies of the 5,5'-dithiobis(2-nitrobenzoic acid) reaction with enzyme I of the phosphoenolpyruvate:glycose phosphotransferase system.
Enzyme I of the bacterial phosphotransferase system is a protein component which undergoes a temperature-dependent monomer/dimer equilibrium. Reaction of sulfhydryl residues with SH-specific reagents inhibits both activity and dimerization. There are four cysteine residues available in each subunit, one of which (Cys 502) is proximate to one of the two tryptophan residues (Trp 498). Previous studies revealed two major lifetimes and spectra, suggesting distinct environments for tryptophan. In this paper, we examine the dynamic quenching of tryptophanyl fluorescence that occurs when an energy transfer acceptor, thio-2-nitrobenzoic acid (TNB), is covalently attached to the sulfhydryl groups. More precisely, we have traced the recovery of nativelike fluorescence lifetime components (and the concomitant loss of "reduced lifetime" amplitudes) that accompanies TNB release. The course of lifetime changes seen when a reducing reagent removes the quencher may be sensitive to a variety of effects, including different SH affinities, different proximities to Trp, changing availability for dimerization, or conformational changes. The prospective value of separating each lifetime component from the mixture is illustrated.


Objective measures of swallowing in patients with primary Sjogren's syndrome.
Primary Sjogren's syndrome (SS) is an autoimmune disorder primarily affecting salivary and lacrimal glands. Durational measures of the oral phase of swallowing were obtained on 34 patients with primary SS and 34 age-matched controls from analyses of ultrasound scans. Two conditions were examined: a basal (BA) swallow (only endogeneous secretions present in the subjects' mouths) and a 10 ml water bolus (WB) swallow. The patients with SS produced swallowing durations significantly longer (p less than 0.05) than those of the controls for each of the two conditions. Moreover, unlike normals, over 40% of the patients with SS produced WB swallows that were longer than their BA swallows. For further analyses, patients with SS were classified into two groups based on the difference in duration between their BA and WB swallows. These two groups differed from each other on clinical evaluations of oral motor function and presenting complaints. No significant differences were found between these two groups for salivary function or immunologic profile. These findings support the hypothesis that dysphagia can result from conditions leading to salivary gland dysfunction and document the need for the assessment of swallowing function in patients with Sjogren's syndrome.


PCr overshoot': a study of the duration in canine myocardium.
The phosphocreatine (PCr) overshoot is a well-documented phenomenon and is readily observable by 31P MRS. In addition, a second 31P MRS observation during ischemia with reperfusion is a diminution in ATP levels. Combining these two as the 'PCr Overshoot' the PCr/ATP ratio may provide an index of viability. However little information is available regarding the duration of this 'overshoot'. For this approach to be useful clinically, the duration of this phenomenon must be ascertained. An open chest canine model of 12 min of ischemia followed by reperfusion (6h) was used. A 2 cm surface coil was sutured to the myocardium and spectra were acquired at 4.7 T. Gated spectra were acquired in <2.5 min with an interpulse delay of 5 s. Integrals of the PCr and ATP (beta) resonances were analyzed using a line-fitting routine. Overall, the PCr signal increased from 22.0+/-0.8 to 25.5+/-0.9 and ATP decreased from 11.7+/-0.4 to 10.0+/-0.4 (arbitrary units). The PCr remained elevated for the entire 6h period and the percentage increase was 15.9%. The ATP remained depleted for the entire 6h period and the percentage decrease was 17.0%. Thus, the clinically relevant and readily observable PCr/ATP is a product of both an increase in PCr and a decrease in ATP for a calculated net increase in PCr/ATP of 39.6%. The PCr/ATP ratio of the ischemia group for baseline, ischemia, 6h reflow, were: 2.33+/-0.18, 1.04+/-0.29 and 3.22+/-0.21. We demonstrate that the 'PCr overshoot' is readily observable and can be monitored noninvasively and nondestructively for 6h. Therefore, the 'PCr overshoot' may be a viable marker of reversible injury in this model and may prove to be applicable for detecting myocardial viability in patients.


Percutaneous placement of two single-lumen Silastic catheters for long-term venous access. Lessons learned with experience in over 200 patients.
A simple, fast technique for percutaneous insertion of two single-lumen, large-bore Silastic catheters for long-term venous access is described. Helpful technical points are discussed. Recent experience with insertion of 102 catheters using this technique yielded no major complications.