Challenges in interpretation of thyroid hormone test results.
In interpreting thyroid hormones results it is preferable to think of interference and changes in concentration of their carrier proteins. We present two patients with discrepancy between the results of thyroid function tests and clinical status. The first case presents a 62-year-old patient with a nodular goiter and Hashimoto thyroiditis. Thyroid function test showed low thyroid-stimulating hormone (TSH) and normal to low fT4. By determining thyroid status (TSH, T4, fT4,T3, fT3) in two laboratories, basal and after dilution, as well as thyroxine-binding globulin (TBG), it was concluded that the thyroid hormone levels were normal. The results were influenced by heterophile antibodies leading to a false lower TSH level and suspected secondary hypothyroidism.The second case, a 40-year-old patient, was examined and followed because of the variable size thyroid nodule and initially borderline elevated TSH, after which thyroid status showed low level of total thyroid hormones and normal TSH. Based on additional analysis it was concluded that low T4 and T3 were a result of low TBG. It is a hereditary genetic disorder with no clinical significance. Erroneous diagnosis of thyroid disorders and potentially harmful treatment could be avoided by proving the interference or TBG deficiency whenever there is a discrepancy between the thyroid function results and the clinical picture.


HIVed, a knowledgebase for differentially expressed human genes and proteins during HIV infection, replication and latency.
Measuring the altered gene expression level and identifying differentially expressed genes/proteins during HIV infection, replication and latency is fundamental for broadening our understanding of the mechanisms of HIV infection and T-cell dysfunction. Such studies are crucial for developing effective strategies for virus eradication from the body. Inspired by the availability and enrichment of gene expression data during HIV infection, replication and latency, in this study, we proposed a novel compendium termed HIVed (HIV expression database; http://hivlatency.erc.monash.edu/) that harbours comprehensive functional annotations of proteins, whose genes have been shown to be dysregulated during HIV infection, replication and latency using different experimental designs and measurements. We manually curated a variety of third-party databases for structural and functional annotations of the protein entries in HIVed. With the goal of benefiting HIV related research, we collected a number of biological annotations for all the entries in HIVed besides their expression profile, including basic protein information, Gene Ontology terms, secondary structure, HIV-1 interaction and pathway information. We hope this comprehensive protein-centric knowledgebase can bridge the gap between the understanding of differentially expressed genes and the functions of their protein products, facilitating the generation of novel hypotheses and treatment strategies to fight against the HIV pandemic.


The concept of coherence in attachment interviews: comparing attachment experts, linguists, and non-experts.
Coherence is a central construct in attachment interviews. Nevertheless, the concept has never been the main focus of a study in the attachment field. The present study examined whether coherence in attachment interviews is defined differently by experts trained in attachment theory, by linguists, and by non-experts. The 72-item Coherence Q-sort (CQS) was used to determine the profile of a prototypical coherent interview. Results indicated that attachment experts could be reliably distinguished from the (combined) other groups: attachment experts emphasized quality and manner more than all other groups, linguists emphasized quantity and relevance more than attachment experts, and higher educated non-experts valued relevance more than attachment experts. Defining coherence in attachment interviews is thus more than just applying Grice's linguistic maxims; expertise in attachment theory is critical for defining interview coherence. Consequences for the coding of the AAI by non-attachment experts, as well as computer coding (im)possibilities are discussed.


Acute Myeloid Leukemia Transformation from Myelofibrosis Upon Remission of an Orbital Myeloid Sarcoma - A Case Report.
To report a case of an orbital myeloid sarcoma concurrent with JAK2 mutation myelofibrosis, which rapidly transformed into acute myeloid leukemia upon aggressive treatment. A 51-year-old woman had progressive swelling of periorbita for one month. Magnetic resonance imaging demonstrated a well-defined, mild enhanced mass indenting the adjacent right lateral rectus muscle and the globe. Biopsy from anterior orbitotomy revealed an orbital myeloid sarcoma. Bone marrow study showed concurrent myelofibrosis. Although the orbital lesion subsided remarkably under aggressive chemotherapy and radiotherapy, the leukemic transformation was noticed in the third month following the initial presentation. This case demonstrated that myeloid sarcoma should be included in the differential diagnosis of orbital diseases, with or without involvement of hematological disorders. Early diagnosis and aggressive treatment as for AML are crucial as the prognosis is usually poor for adult orbital MS.


Adenosine receptor ligands and dizocilpine-induced antinociception in mice.
Interactions between adenosine receptor ligands and dizocilpine (uncompetitive NMDA receptor antagonist) was studied in antinociceptive, writhing test in mice. Minimal effective, antinociceptive doses of adenosine receptor agonists were: 0.1 mg/kg (NECA--A1/A2 agonist). Generally, these agonists did not potentiate the subthreshold dose of dizocilpine (0.05 mg/kg). Of all adenosine receptor antagonists used, only caffeine (A2 and A2 antagonists) reversed dizocilpine-induced (0.1 mg/kg) antinociception dose-dependently. These findings indicate that dizocilpin-induced antinociception in the writhing test is only partly influenced by adenosine receptor ligands.


Is primary anastomosis safe in the surgical management of complications of acute diverticulitis?
The purpose of this article is to determine whether primary anastomosis is a safe option in the surgical management of complications of acute diverticulitis in low-risk patients. Over the past century, the management of diverticulitis has evolved from a three-stage procedure to resection and primary anastomosis. In the beginning of the century, Mayo described drainage and proximal colostomy, a three-stage procedure. This was done by performing a diverting colostomy but leaving the diseased segment of colon, hoping that the inflammation would subside. Later, the patient went back for resection of the diseased segment. Then a third procedure was performed for reversal of the colostomy. Around the late 1970s to early 1980s, it was found that patients had better outcomes if the diseased segment was resected during the first operation-the Hartman procedure. During the late 1990s to early 2000s, some surgeons began performing resection and primary anastomosis in selected groups of patients with diverticulitis. There have been a number of studies published showing that resection and primary anastomosis has an acceptable morbidity and mortality. However, most of these studies are retrospective and do not achieve statistical significance. They also do not attempt to establish guidelines to help decide which patients are good candidates for resection and primary anastomosis. The goal of this study is to establish safe and reasonable practice guidelinesthat can be applied to a selected group of (low-risk) patients. This study is a retrospective review of all the patients treated surgically for complications of acute diverticulitis from 1998 to 2003 at United Hospital Medical Center in Port Chester, New York. Patients were classified as high or low risk based on their age, APACHE II score, American Society of Anesthesiologists class, and Hinchey score. There were a total of 66 patients operated on for complications of acute diverticulitis (left-sided) over this 5-year period. Thirty-six of them underwent resection and primary anastomosis and 30 underwent the Hartman procedure. Of the 36 who underwent resection and primary anastomosis, 19 were considered low risk. There were no complications in this low-risk group who underwent primary anastomosis. Patients who were low risk based on the mentioned criteria can safely undergo resection and primary anastomosis.


Very early onset and greater vulnerability in schizophrenia: A clinical and neuroimaging study.
Although schizophrenia has been diagnosed in children, this group of disorders has received too little attention in the clinical and research literature. Preliminary data suggest that early onset schizophrenia (EOS) and very early onset schizophrenia (VEOS) tend to have a worse outcome than adult onset schizophrenia, and seem to be related to a greater familial vulnerability, due to genetic, psychosocial, and environmental factors. Recently, advanced neuroimaging techniques have revealed structural and functional brain abnormalities in some cerebral areas. This paper reports on a case diagnosed as VEOS, with premorbid year-long psychopathological history. The patient showed atypical proton magnetic resonance spectroscopy findings, and normal brain and spine computer tomography and brain magnetic resonance images.


Canine inflammatory bowel disease: retrospective analysis of diagnosis and outcome in 80 cases (1995-2002).
The case records of 80 dogs in which idiopathic inflammatory bowel disease (IBD) had been diagnosed were reviewed, and owners were contacted for follow-up information using a telephone questionnaire. The types of IBD encountered were lymphocytic (n=6), lymphocytic-plasmacytic (n=38), eosinophilic (n=6) and mixed inflammation (n=30). Prednisolone, sulphasalazine, metronidazole and tylosin were the most frequently prescribed medications. At follow-up, 21 dogs (26 per cent) were classified as being in remission (for a median of 14 months), 40 dogs (50 per cent) had intermittent clinical signs (for a median of 17 months) and three dogs (4 per cent) had uncontrolled disease (for a median of 19 months). Ten dogs (13 per cent) had been euthanased due to refractory IBD and four of these had entered remission for a median of 21 months prior to developing severe relapse and refractoriness to further treatment. Six dogs (8 per cent) had been euthanased or had died for reasons unrelated to IBD. Hypoalbuminaemia at the time of diagnosis was significantly associated with a negative outcome (P=0.0007). No association was found between the site (P=0.75), type (P=0.44) and severity (P=0.75) of disease. Dietary change to single protein and carbohydrate commercial diets had no association with outcome (P=0.12). Owner assessment of quality of life at follow-up was significantly associated with outcome (P=0.006).


Comprehensive study of gene and microRNA expression related to epithelial-mesenchymal transition in prostate cancer.
Prostate cancer is the most common cancer in men, and most patients have localized disease at the time of diagnosis. However, 4% already present with metastatic disease. Epithelial-mesenchymal transition is a fundamental process in carcinogenesis that has been shown to be involved in prostate cancer progression. The main event in epithelial-mesenchymal transition is the repression of E-cadherin by transcription factors, but the process is also regulated by microRNAs. The aim of this study was to analyze gene and microRNA expression involved in epithelial-mesenchymal transition in localized prostate cancer and metastatic prostate cancer cell lines and correlate with clinicopathological findings. We studied 51 fresh frozen tissue samples from patients with localized prostate cancer (PCa) treated by radical prostatectomy and three metastatic prostate cancer cell lines (LNCaP, DU145, PC3). The expression of 10 genes and 18 miRNAs were assessed by real-time PCR. The patients were divided into groups according to Gleason score, pathological stage, preoperative PSA, biochemical recurrence, and risk group for correlation with clinicopathological findings. The majority of localized PCa cases showed an epithelial phenotype, with overexpression of E-cadherin and underexpression of the mesenchymal markers. MiRNA-200 family members and miRNAs 203, 205, 183, 373, and 21 were overexpressed, while miRNAs 9, 495, 29b, and 1 were underexpressed. Low-expression levels of miRNAs 200b, 30a, and 1 were significantly associated with pathological stage. Lower expression of miR-200b was also associated with a Gleason score ≥ 8 and shorter biochemical recurrence-free survival. Furthermore, low-expression levels of miR-30a and high-expression levels of Vimentin and Twist1 were observed in the high-risk group. Compared with the primary tumor, the metastatic cell lines showed significantly higher expression levels of miR-183 and Twist1. In summary, miRNAs 200b, 30a, 1, and 183 and the genes Twist1 and Vimentin might play important roles in the progression of prostate cancer and may eventually become important prognostic markers.


Women with painful breasts without palpable masses: do they really need a mammogram?
This prospective study was carried out over a period of 36 months, and the study population was, all the women (726) referred for mammograms on account of painful breast(s) without palpable masses. They were matched with the same number of asymptomatic women in a control group. The mammographic findings in the women with painful breast(s) and no palpable masses were normal in 639 (88%), benign in 80 (11%), suspicious in 2 (0.3%) and malignant in 5 (0.7%). In the control group mammograms showed normal breasts in 625 (86.1%), benign and suspicious lesions in 87 (12%) and 7 (1.0%) respectively. Malignant lesions were recorded in 7 (0.9%) women. The prevalence of breast cancer was similar in women with painful breast(s) and the control asymptomatic cases. Our reports show that inspite of the pain experienced by the women in this study mammography had a low diagnostic yield of malignant lesions, just as it was found in the control group. Mammography therefore, in these patients will only provide reassurance.


Islet amyloid polypeptide and insulin relationship in a longitudinal study of the genetically obese (ob/ob) mouse.
Obese mice (Umeå ob/ob) and their lean litter-mates were investigated from 7 to 52 weeks of age with respect to the plasma concentration of islet amyloid polypeptide (IAPP) and insulin. Plasma levels of IAPP were highly elevated in the ob/ob mice and remained unchanged until age 33 weeks, after which a sudden significant increase occurred at age 40 weeks. The plasma concentration of insulin gradually increased from start to end and reached extremely high levels. In the lean mice, there were no age-related differences in plasma levels of IAPP and insulin, being of the same magnitude as in normal NMRI mice. The plasma IAPP/insulin molar ratio was similar in lean and obese mice until age 14 weeks. At 21 weeks, the ratio in the ob/ob mice had decreased dramatically and remained markedly (sixfold) lower than in the lean mice until the end of the study. The IAPP concentration in the pancreata of 21-week-old ob/ob mice was 25-fold higher than that in the lean mice. Immunohistochemically, a majority of the ob/ob mice displayed enlarged and more numerous pancreatic islets, compared with the lean mice, and the IAPP- and insulin-labeling intensity was equal for all animals. At the electron-microscopic level, there was an increase in the number of IAPP- and insulin-immunoreactive gold particles per whole granule area as well as per core granule area. We conclude that the dramatically increased IAPP levels in severely hyperinsulinemic ob/ob mice may be of importance for the development of insulin resistance. Further, the disproportionate secretion of IAPP and insulin in the adult obese mouse might indicate a disturbed negative feedback effect of IAPP on insulin secretory mechanisms, resulting in very high plasma insulin levels.


Return-to-play guidelines in concussion: revisiting the literature.
Return-to-play (RTP) guidelines in concussion management and treatment continue to challenge clinicians, despite recent updates to the protocols and the ongoing media attention. The current guidelines address individualized treatment planning but are difficult to apply to athletes who experience prolonged symptoms, are younger, or have sustained multiple concussions. Recent literature findings have contributed to an improved understanding and applicability of the guidelines while emphasizing a multidisciplinary paradigm in formulating an individualized RTP decision. Successful implementation of the RTP guidelines will ensure that athletes are protected from further injury and return to their baseline functional status.


[Expression of hepatitis B virus antigen in brain tissue from liver cirrhosis patients with hepatitis B and its significance].
To investigate the expression of hepatitis B virus (HBV) antigen in brain tissue from liver cirrhosis patients with hepatitis B and explore its significance. HBsAg and HBcAg were detected in the brain tissue from 70 liver cirrhosis dead patients with hepatitis B by S-P immunohistochemical assay, and the relationship between the expression of HBV antigen in brain tissue testing and clinic and pathology was analyzed. 30 patients (42.89%) were positive for HBV antigen. Among them, 24 patients (34.29%) were positive for HBsAg and 18 patients (25.71%) were positive for HBcAg. HBV antigen was mainly found in cytoplasm and distributed in neurons, neurogliocytes and vascular endothelial cells. The positive cells were distributed separately, scatteredly or focally. The expression of HBV antigen was not associated with the serum level of HBV replication but associated with the occurrence of hepatic encephalopathy (HE) and the severity of brain tissue pathologic lesions of HE. The results indicate that HBV infection develop in brain tissue from liver cirrhosis patients with hepatitis B and HBV may replicate in it. HBV infection in brain tissue may p lay an important role in occurrence and development of HE from liver cirrhosis patients with hepatitis B.


[Cervical pregnancy].
The cervical pregnancy is one of the states that make the direct life-threatening for a woman. A death rate is about 20-60%. It is estimated that the cervical pregnancy occurs once for 16000 cases of normal pregnancy. Etiology is not recognized well yet. It is assumed that one of the major causes for implantation of fetal egg within a uterine cervix is the decidual metabolism of endometrium. A hyperestrogenic period after fertilization causing a change of oviduct kinetics may be conducive to nidate the egg in the cervical canal. Surgical traumas of uterus, multiparity, development anomaly, tumours of reproductive organs may cause the change of the fetal egg localization. The great significance is attributed recently to an atrophy of transparent tunica and involving a retardation of egg innidiation ability what results in localization of pregnancy in the cervical canal. In case of this type of ectopic pregnancy the only way to recognize it is a precise medical history of a patient, clinical picture and gynaecological examination. An early recognition of the cervical pregnancy prevents of metrorrhagia which is so dangerous for life. It is commonly assumed that a therapy to be chosen is removal of uterus. Using this background one case of the cervical pregnancy observed for the last 20 years in the department materials is described which recognition before treatment was settled based on the presented clinical argumentation.


Hormone replacement therapy and breast cancer risk in Kyushu University Hospital: supporting the Women's Health Initiative study.
The influence of hormone replacement therapy (HRT) on breast cancer has not been clarified in Japan, however the Women's Health Initiative (WHI) trial recently showed breast cancer risk according to use of estrogen plus progestin. We assessed retrospectively the incidence of breast cancer in postmenopausal women who received HRT at our outpatients clinic. Among the patients registered at the postmenopausal clinic in Kyushu University Hospital from 1990 to 2003, 917 women who received HRT (estrogen plus progestin, 507 patients; estrogen only, 410 patients) after informed consent were examined by mammography or breast ultrasound tomography. Breast cancer occurred in nine patients: five patients who received estrogen plus progestin and four who received estrogen only. Four of five the patients who used estrogen plus progestin received HRT for more than 5 years. Breast cancer risk in patients who used estrogen plus progestin for more than 5 years seemed to be higher than those who used it for less than 5 years. This tendency is similar to the results of the WHI. In addition, breast cancer incidence in patients treated with HRT in our clinic seemed to be higher than the incidence in average Japanese women. Therefore, patients need to be given sufficient information before undergoing HRT.


Unwound regions in yeast centromere IV DNA.
The DNA of the centromere of chromosome IV (CEN4) of Saccharomyces cerevisiae is found to be sensitive to single-strand specific nuclease P1 when inserted into a negatively supercoiled plasmid. Fine mapping identifies two P1-sensitive segments: one segment maps to essential centromere element CDEI and bordering CDEII bases, and the other segment is located in element CDEIII. The AT-rich element CDEII, which is expected to be early melting, is for the most part resistant to nuclease P1. Cleavage is inhibited by NaCl, MgCl2 and polyamines. The cleavage rate is only weakly dependent on P1 concentration in the range of 0.5 to 20 munits/microliters. The two P1-sensitive segments are also modified by the DNA-confirmation-specific reagent KMnO4. Negative superhelicity is required for all modifications. Two-dimensional topoisomer analysis indicates the unwinding of 80(+/- 10) bases within the negatively supercoiled CEN4-containing plasmid. The data best fit a model in which the DNA of the CEN4 region undergoes a transition into a paranemic intermediate in which each strand is folded into an RNA-like foldback structure.


Wrong postural hygiene and ergonomics in dental students of the University of Valencia (Spain) (part I).
Failure to adopt a correct working posture can lead to occupational diseases. Evaluate knowledge in relation to ergonomics about BHOP concept and its application to routine clinical practice amongst undergraduate and postgraduate dental students in the University of Valencia (Valencia, Spain). A study based on interviews of undergraduate and postgraduate dental students in the University of Valencia (Valencia, Spain) was carried out. The information from a total of 336 interviews was used for the statistical analysis, differentiating according to gender and academic year: knowledge of ergonomics, pain prevalence and antecedents, assessment of the possible necessity for improved training in ergonomics, and evaluation of postural hygiene. Only 28.6% of the students were found to sit correctly in the dentist chair. Furthermore, in the opinion of the students, very few subjects during the career afforded adequate teaching in relation to ergonomics and working posture. The analysis of postural hygiene showed great variability. There were no significant differences in posture between males and females, although some incorrect postures appeared to be associated with the academic year (P<.05). However, no significant improvements in postural hygiene were noted on progressing from one academic year to the next. The students in our study were not familiar with the principles of ergonomics and did not sit correctly in the dentist chair. Improved training in this field is required in dental school.


Cysteine Oxidations in Mitochondrial Membrane Proteins: The Case of VDAC Isoforms in Mammals.
Cysteine residues are reactive amino acids that can undergo several modifications driven by redox reagents. Mitochondria are the source of an abundant production of radical species, and it is surprising that such a large availability of highly reactive chemicals is compatible with viable and active organelles, needed for the cell functions. In this work, we review the results highlighting the modifications of cysteines in the most abundant proteins of the outer mitochondrial membrane (OMM), that is, the voltage-dependent anion selective channel (VDAC) isoforms. This interesting protein family carries several cysteines exposed to the oxidative intermembrane space (IMS). Through mass spectrometry (MS) analysis, cysteine posttranslational modifications (PTMs) were precisely determined, and it was discovered that such cysteines can be subject to several oxidization degrees, ranging from the disulfide bridge to the most oxidized, the sulfonic acid, one. The large spectra of VDAC cysteine oxidations, which is unique for OMM proteins, indicate that they have both a regulative function and a buffering capacity able to counteract excess of mitochondrial reactive oxygen species (ROS) load. The consequence of these peculiar cysteine PTMs is discussed.


32P-postlabeling assay for free radical-induced DNA damage: the formamido remnant of thymine.
A prominent lesion in DNA exposed to oxidative free radicals results from the degradation of thymine leaving a formamido remnant. A 32P-postlabeling assay has been developed for the detection of the formamido lesion. The assay is based on the circumstance that the lesion prevents hydrolysis by nuclease P1 of the phosphoester bond 3' to the damaged nucleoside. Thus, a nuclease P1 plus acid phosphatase digest of DNA generates mostly nucleosides whereas the formamido lesion is rendered as a modified dinucleoside monophosphate. Dinucleoside monophosphates, but not nucleosides, are apt substrates for 32P-postlabeling by polynucleotide kinase. The assay was applied to calf thymus DNA X-irradiated in oxygenated solution. The formamido lesion could be detected down to a dose of a few Gy.


Antimicrobial activity of a new synthetic peptide loaded in polylactic acid or poly(lactic-co-glycolic) acid nanoparticles against Pseudomonas aeruginosa, Escherichia coli O157:H7 and methicillin resistant Staphylococcus aureus (MRSA).
Nanocarrier systems are currently being developed for peptide, protein and gene delivery to protect them in the blood circulation and in the gastrointestinal tract. Polylactic acid (PLA) and poly(lactic-co-glycolic) acid (PLGA) nanoparticles loaded with a new antimicrobial GIBIM-P5S9K peptide were obtained by the double emulsion solvent extraction/evaporation method. PLA- and PLGA-NPs were spherical with sizes between 300 and 400 nm for PLA and 200 and 300 nm for PLGA and <0.3 polydispersity index as determined by dynamic light scattering and scanning electron microscopy), having the zeta potential of >20 mV. The peptide-loading efficiency of PLA-NP and PLGA-NPs was 75% and 55%, respectively. PLA- and PLGA-NPs released around 50% of this peptide over 8 h. In 10% human sera the size of peptide loaded PLA- and PLGA-NPs increased between 25.2% and 39.3%, the PDI changed from 3.2 to 5.1 and the surface charge from -7.15 to 14.6 mV. Both peptide loaded PLA- and PLGA-NPs at 0.5 μM peptide concentration inhibited the growth of Escherichia coli O157:H7 (E. coli O157:H7), methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas. aeruginosa (P. aeruginosa). In contrast, free peptide inhibited at 10 μM but did not inhibit at 0.5 and 1 μM. These PLA- and PLGA-NPs presented <10% hemolysis indicating that they are hemocompatible and promising for delivery and protection system of GIBIM-P5S9K peptide.


Specificity of antibodies to heterologous glomerular and tubular basement membranes in various strains of mice with different H-2 types.
C3H, CBA (H-2k) and NZB (H-2d) mice were immunized with dog insoluble glomerular (GBM) or tubular basement membrane (TBM). The titre of circulating antibodies was sequentially determined and their specificity was analysed using various soluble antigenic fractions. Glomerular and tubular deposits were studied on serial biopsies by direct immunofluorescence. After elution from whole kidneys, IgG fixation on normal mouse kidney sections was analysed by indirect immunofluorescence. After immunization with insoluble GBM, animals from all three strains develop antibodies mainly directed against collagenous antigenic determinants shared by GBM and TBM. After immunization with insoluble TBM, the antibodies are directed in NZB mice against non-collagenous TBM-specific determinants, in C3H mice against collagenous determinants and in CBA mice against both types of antigenic determinants. Thus the ability to respond to the various antigens of GBM and TBM is genetically determined and does not depend only on the major histocompatibility complex.


Targeting the differential addiction to anti-apoptotic BCL-2 family for cancer therapy.
BCL-2 family proteins are central regulators of mitochondrial apoptosis and validated anti-cancer targets. Using small cell lung cancer (SCLC) as a model, we demonstrated the presence of differential addiction of cancer cells to anti-apoptotic BCL-2, BCL-XL or MCL-1, which correlated with the respective protein expression ratio. ABT-263 (navitoclax), a BCL-2/BCL-XL inhibitor, prevented BCL-XL from sequestering activator BH3-only molecules (BH3s) and BAX but not BAK. Consequently, ABT-263 failed to kill BCL-XL-addicted cells with low activator BH3s and BCL-XL overabundance conferred resistance to ABT-263. High-throughput screening identified anthracyclines including doxorubicin and CDK9 inhibitors including dinaciclib that synergized with ABT-263 through downregulation of MCL-1. As doxorubicin and dinaciclib also reduced BCL-XL, the combinations of BCL-2 inhibitor ABT-199 (venetoclax) with doxorubicin or dinaciclib provided effective therapeutic strategies for SCLC. Altogether, our study highlights the need for mechanism-guided targeting of anti-apoptotic BCL-2 proteins to effectively activate the mitochondrial cell death programme to kill cancer cells.


Unexpected effect of catalyst concentration on photochemical CO2 reduction by trans(Cl)-Ru(bpy)(CO)2Cl2: new mechanistic insight into the CO/HCOO- selectivity.
Photochemical CO2 reduction catalysed by trans(Cl)-Ru(bpy)(CO)2Cl2 (bpy = 2,2'-bipyridine) efficiently produces carbon monoxide (CO) and formate (HCOO-) in N,N-dimethylacetamide (DMA)/water containing [Ru(bpy)3]2+ as a photosensitizer and 1-benzyl-1,4-dihydronicotinamide (BNAH) as an electron donor. We have unexpectedly found catalyst concentration dependence of the product ratio (CO/HCOO-) in the photochemical CO2 reduction: the ratio of CO/HCOO- decreases with increasing catalyst concentration. The result has led us to propose a new mechanism in which HCOO- is selectively produced by the formation of a Ru(i)-Ru(i) dimer as the catalyst intermediate. This reaction mechanism predicts that the Ru-Ru bond dissociates in the reaction of the dimer with CO2, and that the insufficient electron supply to the catalyst results in the dominant formation of HCOO-. The proposed mechanism is supported by the result that the time-course profiles of CO and HCOO- in the photochemical CO2 reduction catalysed by [Ru(bpy)(CO)2Cl]2 (0.05 mM) are very similar to those of the reduction catalysed by trans(Cl)-Ru(bpy)(CO)2Cl2 (0.10 mM), and that HCOO- formation becomes dominant under low-intensity light. The kinetic analyses based on the proposed mechanism could excellently reproduce the unusual catalyst concentration effect on the product ratio. The catalyst concentration effect observed in the photochemical CO2 reduction using [Ru(4dmbpy)3]2+ (4dmbpy = 4,4'-dimethyl-2,2'-bipyridine) instead of [Ru(bpy)3]2+ as the photosensitizer is also explained with the kinetic analyses, reflecting the smaller quenching rate constant of excited [Ru(4dmbpy)3]2+ by BNAH than that of excited [Ru(bpy)3]2+. We have further synthesized trans(Cl)-Ru(6Mes-bpy)(CO)2Cl2 (6Mes-bpy = 6,6'-dimesityl-2,2'-bipyridine), which bears bulky substituents at the 6,6'-positions in the 2,2'-bipyridyl ligand, so that the ruthenium complex cannot form the dimer due to the steric hindrance. We have found that this ruthenium complex selectively produces CO, which strongly supports the catalytic mechanism proposed in this work.


Intracranial subdural empyema after surgery for lumbar lipomyelomeningocele: A rare complication.
Surgery is routinely recommended for lumbar lipomyelomeningocele, especially in the setting of tethered cord syndrome. The most common complications are wound infections and cerebrospinal fluid (CSF) leak, which remain confined to the surgical site. To the best of our knowledge, there have been no prior reports relating an intracranial subdural empyema following detethering surgery. Prompt diagnosis is essential since subdural empyema is a neurosurgical emergency. The patient was an 11-month-old male who underwent detethering surgery for a lumbar lipomyelomeningocele. This was followed by wound drainage consistent with CSF leak, requiring revision. Cultures grew three aerobes (Escherichia coli, Enterococcus, and Klebsiella) and three anaerobes (Clostridium, Veillonella, and Bacteroides). He was started on cefepime, vancomycin, and flagyl. The patient required two more wound revisions and placement of an external ventricular drain (EVD) secondary to persistent wound leakage. A subsequent magnetic resonance imaging (MRI) brain was carried out due to protracted irritability, which revealed extensive left subdural empyema along the parietooccipital region and the inferior and anterior temporal lobe. He underwent evacuation of the subdural empyema where cultures exhibited no growth. Subsequently, he progressed well. His lumbar incision continued to heal. Serial MRI brains and inflammatory markers were reassuring. He weaned off his EVD and went home to complete a 6-week course of antibiotics. Upon completion of his antibiotics, he returned for a clinic visit; he exhibited no interim fevers or wound issues; cranial imaging documented no evidence of a residual or recurrent subdural empyema. Intracranial subdural empyema may occur after wound complications from detethering surgery despite early initiation of broad-spectrum antibiotics. Possible etiology may be local wound infection that seeds the subdural space and travels to the cranium, leading to meningitis and subdural empyema. Such a scenario should prompt surveillance imaging of the head as undiagnosed subdural empyema may lead to devastating consequences.


Exercise Modality Effect on Bioenergetical Performance at V˙O2max Intensity.
A bioenergetical analysis of different exercise modes near maximal oxygen consumption (V˙O2max) intensity is scarce, hampering the prescription of training to enhance performance. We assessed the time sustained in swimming, rowing, running, and cycling at an intensity eliciting V˙O2max and determined the specific oxygen uptake (V˙O2) kinetics and total energy expenditure (Etot-tlim). Four subgroups of 10 swimmers, 10 rowers, 10 runners, and 10 cyclists performed (i) an incremental protocol to assess the velocity (vV˙O2max) or power (wV˙O2max) associated with V˙O2max and (ii) a square wave transition exercise from rest to vV˙O2max/wV˙O2max to assess the time to voluntary exhaustion (Tlim-100%V˙O2max). The V˙O2 was measured using a telemetric portable gas analyzer (K4b, Cosmed, Rome, Italy) and V˙O2 kinetics analyzed using a double exponential curve fit. Etot-tlim was computed as the sum of its three components: aerobic (Aer), anaerobic lactic (Analac), and anaerobic alactic (Anaalac) contributions. No differences were evident in Tlim-100% V˙O2max between exercise modes (mean ± SD: swimming, 187 ± 25; rowing, 199 ± 52; running, 245 ± 46; and cycling, 227 ± 48 s). In contrast, the V˙O2 kinetics profile exhibited a slower response in swimming (21 ± 3 s) compared with the other three modes of exercise (rowing, 12 ± 3; running, 10 ± 3; and cycling, 16 ± 4 s) (P < 0.001). Etot-tlim was similar between exercise modes even if the Analac contribution was smaller in swimming compared with the other sports (P < 0.001). Although there were different V˙O2 kinetics and ventilatory patterns, the Tlim-100%V˙O2max was similar between exercise modes most likely related to the common central and peripheral level of fitness in our athletes.


Medical costs of a low skeletal muscle mass are modulated by dietary diversity and physical activity in community-dwelling older Taiwanese: a longitudinal study.
Age-related loss of skeletal muscle mass (SMM) and function (sarcopenia) are associated with poor health outcomes and an economic burden on health care services. An appropriate diet and physical activity have been proposed for prevention and treatment of sarcopenia. Nevertheless, the effects on medical service utilization and costs remain unclear. This study determined the effects of SMM in conjunction with diet quality and physical activity on medical service utilization and expenditure in community-dwelling older Taiwanese. In total, 1337 participants from the Elderly Nutrition and Health Survey in Taiwan (1999-2000) were enrolled. An SMM index [SMMI, calculated by dividing SMM (kg) by height (m2)] was used as the marker of sarcopenia. Participants with the lowest SMMI quartiles (<11.4 kg/m2 for men and 8.50 kg/m2 for women) comprised the high-risk group, and the remainder comprised the low-risk group. Dietary information (dietary diversity: low and high) and physical activity (low and moderate) were obtained at baseline. Annual medical service utilization and expenditure were calculated from National Health Insurance claims until December 31, 2006. Generalized linear models were used to determine the association between the SMMI and annual medical service utilization and costs in conjunction with dietary diversity or physical activity. After 8 follow-up years, regardless of gender, participants in the high-risk group reported significantly more hospitalization (days and expenditure) and total medical expenditure. Participants in the high-risk group who had low dietary diversity made fewer annual outpatient (14%), preventive care (19%), and dental (40%) visits, but exhibited longer hospitalization (102%) than did those who had a low SMMI and high dietary diversity. Similar patterns were observed in the corresponding medical expenditures. The findings were similar when considering physical activity. Being in the low-risk group in conjunction with having high dietary diversity or more physical activity was associated with the lowest annual adjusted mean hospitalization days with expenditure, and also total expenditure. A lower SMMI was associated with more hospitalization days and costs. However, high dietary diversity and more physical activity can attenuate the effects of lower SMMI on medical service utilization and expenditure.


Renal handling of NH4+ in relation to the control of acid-base balance by the kidney.
The major component of urinary acid excretion is NH4+. To be appropriately excreted in urine, NH4+ must be synthesized by proximal tubular cells, secreted into the proximal tubular fluid, reabsorbed by the medullary thick ascending limb (MTAL) to be accumulated in the medullary interstitium, and finally secreted in medullary collecting ducts. Each step of this renal pathway is highly regulated and, in addition to acute events mediated by peptide hormones such as parathyroid hormone, the control of gene expression explains how the renal handling of NH4+ fully adapts to chronic changes in the acid-base status. Several targets have been identified at the gene expression level to account for the adaptation of renal NH4+ synthesis and transport in response to an acid load. These are the key enzymes of ammoniagenesis (mitochondrial glutaminase and glutamate dehydrogenase) and gluconeogenesis (phosphoenolpyruvate carboxykinase) in the proximal tubule, the apical Na(+)-K+(NH4+)-2Cl- cotransporter of the MTAL, and the basolateral Na(+)-K+(NH4+)-2Cl- cotransporter of medullary collecting ducts. At least two factors control the expression of these genes during metabolic acidosis: an acid pH and glucocorticoids, which appear to act in concert to coordinate the adaptation of various tubular cell types. The present review focuses on some aspects of these regulations that have been recently elucidated.


Late Pleistocene/Early Holocene Migratory Behavior of Ungulates Using Isotopic Analysis of Tooth Enamel and Its Effects on Forager Mobility.
Zooarchaeological and paleoecological investigations have traditionally been unable to reconstruct the ethology of herd animals, which likely had a significant influence on the mobility and subsistence strategies of prehistoric humans. In this paper, we reconstruct the migratory behavior of red deer (Cervus elaphus) and caprids at the Pleistocene-Holocene transition in the northeastern Adriatic region using stable oxygen isotope analysis of tooth enamel. The data show a significant change in δ18O values from the Pleistocene into the Holocene, as well as isotopic variation between taxa, the case study sites, and through time. We then discuss the implications of seasonal faunal availability as determining factors in human mobility patterns.


Chikungunya as a paradigm for emerging viral diseases: Evaluating disease impact and hurdles to vaccine development.
Chikungunya fever (CHIKF) is an emerging infectious disease caused by an alphavirus transmitted by Aedes spp. mosquitoes. Because mosquito control programs are not highly efficient for outbreak containment, vaccines are essential to reduce the burden of disease. Although no licensed vaccine against CHIKF is yet available, many highly promising candidates are undergoing preclinical studies, and a few of them have been tested in human trials of phase 1 or 2. Here, we review recent findings regarding the need for a CHIKF vaccine and provide an update on vaccines nearing or having entered clinical trials. We also address needs to tackle bottlenecks to vaccine development-including scientific and financial barriers-and to accelerate the development of vaccines; several actions should be taken: (i) design efficacy trials to be conducted during the course of outbreaks; (ii) evaluate the opportunity for adopting the "animal rule"for demonstration of efficacy for regulatory purposes; (iii) strengthen the collective commitment of nations, international organizations, potential donors and industry; (iv) stimulate public and/or private partnerships to invest in vaccine development and licensure; and (v) identify potential markets for an effective and safe CHIKF vaccine.


Inequity of healthcare utilization on mammography examination and Pap smear screening in Thailand: Analysis of a population-based household survey.
Healthcare in Thailand is not equally distributed, and not all people can equally access healthcare resources even if they are covered by health insurance. To examine factors associated with the utilization of mammography examination for breast cancer and Pap smear screening for cervical cancer, data from the national reproductive health survey conducted by the National Statistical Office of Thailand in 2009 was examined. The survey was carried out on 15,074,126 women aged 30-59 years. The results showed that the wealthier respondents had more mammograms than did the lower-income groups. The concentration index was 0.144. The data on Pap smears for cervical cancer also showed that the wealthier respondents were more likely to have had a Pap smear than their lower-income counterparts. The concentration index was 0.054. Determinants of mammography examination were education, followed by health welfare and wealth index, whereas the determinants of Pap smear screening were wealth index, followed by health welfare and education. The government should support greater education for women because education was associated with socioeconomic status and wealth. There should be an increase in the number of screening campaigns, mobile clinics, and low-cost mammograms and continued support for accessibility to mammograms, especially in rural areas and low-income communities.


Duration of diabetes and carotid wall thickness. The Insulin Resistance Atherosclerosis Study (IRAS).
Diabetes is a major risk factor for morbidity and mortality from cardiovascular disease. However, the role of the primary metabolic abnormality of diabetes (chronic hyperglycemia) in this disease process has not been fully elucidated. A cross-sectional analysis was conducted among 489 persons with non-insulin-dependent diabetes mellitus; 299 were established diabetics (diagnosed previously) and 190 were newly diagnosed at the time of the Insulin Resistance Atherosclerosis Study (IRAS) examination. These men and women, of three different ethnic groups, were participants in IRAS. Established diabetes (versus newly diagnosed diabetes) and mean fasting glucose level were used as measures of hyperglycemic burden. Intimal-medial wall thickness (IMT) of the internal (ICA) and common (CCA) carotid arteries were used as indices of atherosclerosis. The mean duration of disease among established diabetics was 7 years. The mean CCA IMT and ICA IMT were 872 and 946 microns, respectively. Established diabetes and mean fasting glucose level were positively associated with increased CCA IMT (P < .05) but not ICA IMT, even after adjustment for numerous cardiovascular disease risk factors. CCA IMT was increased by 70 microns in established diabetics (versus newly diagnosed diabetics) and by 26 microns per 1 SD of fasting glucose. Among established diabetics, however, duration of known diabetes (number of years) was not significantly related to IMT. Among diabetics in IRAS, established diabetes and fasting glucose level were each independently associated with CCA IMT, suggesting that chronic hyperglycemia or its associated metabolic abnormalities may lead to increased risk of atherosclerosis.


Identification of Hypoderma actaeon (Diptera: Oestridae) in red deer (Cervus elaphus) from northern Spain: Microscopy study and molecular analysis.
Hypoderma spp. larvae were observed subcutaneously in the dorsal and lumbar regions of red deer (Cervus elaphus) hunted in the province of León (northwestern Spain) causing a myiasis. They were removed and initially classified by their size, shape, color, and location under the skin into the three larval stages that parasitize these animals. The morphological characteristics of the first and second-instar are described and from the features of the third-instar the species was identified as Hypoderma actaeon. To accurately identify this species, five isolates of genomic DNA from the third-instar, two from the second-instar and two from first-instar of H. actaeon were analyzed by PCR analysis of the COI region of mt-DNA. The results confirmed that the examined samples exactly matched with H. actaeon. This study has shown the morphological identification of the three larval stages of H. actaeon and, for the first time, the first and second-instar larvae have been molecularly characterized. Finally, identification of only H. actaeon suggests that this species is the only affecting red deer in the Iberian Peninsula.


The hirudin-like factors HLF3 and HLF4-hidden hirudins of European medicinal leeches.
The hirudin-like factors 3 (HLF3) and 4 (HLF4) belong to a new class of leech-derived factors and are present in specimens of the three European medicinal leeches, Hirudo medicinalis, Hirudo verbana, and Hirudo orientalis, respectively. Here we describe the functional analysis of natural and synthetic variants of HLF3 and HLF4. Whereas the natural variants display only very low or no detectable anti-coagulatory activities, modifications within the N-termini in combination with an exchange of the central globular domain have the potency to greatly enhance the inhibitory effects of respective HLF3 and HLF4 variants on blood coagulation. Our results support previous observations on the crucial importance of all parts (both the N- and C-termini as well as the central globular domains) of hirudin and HLF molecules for thrombin inhibition.


Oxidation of Tyrosine-Phosphopeptides by Titanium Dioxide Photocatalysis.
Protein phosphorylation has a key role in cell regulation. Oxidation of proteins, in turn, is related to many diseases and to aging, but the effects of phosphorylation on the oxidation of proteins and peptides have been rarely studied. The aim of this study was to examine the mechanistic effect of phosphorylation on peptide oxidation induced by titanium dioxide photocatalysis. The effect of phosphorylation was compared between nonphosphorylated and tyrosine phosphorylated peptides using electrospray tandem mass spectrometry. We observed that tyrosine was the most preferentially oxidized amino acid, but the oxidation reaction was significantly inhibited by its phosphorylation. The study also shows that titanium dioxide photocatalysis provides a fast and easy method to study oxidation reactions of biomolecules, such as peptides.


In vitro study of Organization for Economic Co-operation and Development (OECD) endocrine disruptor screening and testing methods- establishment of a recombinant rat androgen receptor (rrAR) binding assay.
The androgen receptor (AR) binding assay can be used to determine the ability of probable endocrine disruptors (EDs) to compete with synthetic androgen methyltrienolone (R1881) for binding to recombinant rat AR (rrAR). In this study, we assessed AR binding of various chemicals using Lexius Freyberger's method. The rank of relative binding affinity (RBA, IC(50)) on the tested chemicals was trenbolone 1.3 x 10(-8) M (RBA 138) > dihydrotesterone (DHT) 1.8 x 10(-8) M (RBA 100) > methyl testosterone 5.7 x 10(-8) M (RBA 31.6) > nonylphenol (NP) 1.3 x 10(-5) M (RBA 0.14) > bisphenol A (BPA) 1.1 x 10(-4) M (RBA 0.016) > isobutyl paraben 3.1 x 10(-4) M (RBA 0.0058) > butyl paraben 6.2 x 10(-4) M (RBA 0.0029) > propyl paraben 9.7 x 10(-4) M (RBA 0.0019). However, di(n-butyl) phthalate (DBP) and di(2-ethylhexyl) phthalate (DEHP), known anti-androgenic chemicals, did not show any significant AR binding activity. Our data suggests that in vitro AR binding assay may be useful as a screening tool for potential EDs.


[Pregnancy interruption in Switzerland 1942-1996].
In spite of restrictive legislation, practice of legal abortion has become liberal in Switzerland, bringing illegal abortions to disappear. Moreover, sex education and widespread contraception have reduced the number of legal abortions. Today, in spite of liberalization, the abortion rate is among the lowest worldwide. Prevention targeted specifically at migrant women and spreading knowledge about postcoital contraception might reduce it further. Liberalization in practice must be followed by liberalizing legislation.


[Effects of lead on somatostatin positive neuron and its mRNA expression in rat's hippocampus].
In order to study the changes of hippocampal SS and SSmRNA positive neurons, Wistar rats were exposed to lead by drinking 2000 micrograms/ml lead acetate solution for three months, the blood and hippocampal lead concentration were measured with atomic absorption spectometry, and the number of hippocampal SS and SSmRNA positive neurons were observed with immunohistochemistry, and in situ hybridization respectively. The results showed that the blood and hippocampal lead content in lead-exposed rats were obviously higher than those of the control rats (130.05 +/- 14.21) mg/L and (1011.62 +/- 59.54) ng/L respectively v.s (5.54 +/- 0.76) mg/L and (181.98 +/- 47.27) ng/L respectively (P < 0.05). The number of SS and SSmRNA positive neurons in CA1 area of lead-exposed rats were significantly smaller than those of the control rats (23.27 +/- 4.58 and 37.60 +/- 9.69 respectively v.s. 40.38 +/- 7.61 and 53.48 +/- 10.22 respectively, P < 0.01). The numbers of SS and SSmRNA positive neurons in CA3 area of lead-exposed rats were smaller than that of the controls rats (12.16 +/- 3.40 and 16.18 +/- 6.19 respectively v.s 17.68 +/- 5.86 and 25.93 +/- 7.47 respectively, P < 0.05). The study suggested that the hippocampal SS and SSmRNA positive neurons of lead-exposed rat have a lower expression of SS and SSmRNA.


Mycophenolate mofetil toxicity mimicking acute cellular rejection in a small intestinal transplant.
Mycophenolate mofetil (MMF) is an important medication used for maintenance immunosuppression in solid organ transplants. A common gastrointestinal (GI) side effect of MMF is enterocolitis, which has been associated with multiple histological features. There is little data in the literature describing the histological effects of MMF in small intestinal transplant (SIT) recipients. We present a case of MMF toxicity in a SIT recipient, with histological changes in the donor ileum mimicking persistent acute cellular rejection (ACR). Concurrent biopsies of the patient's native colon showed similar changes to those from the donor small bowel, suggesting a non-graft specific process, raising suspicion for MMF toxicity. The MMF was discontinued and complete resolution of these changes occurred over three weeks. MMF toxicity should therefore be considered as a differential diagnosis for ACR and graft-versus-host disease in SITs.


Accurate Compensation of Attitude Angle Error in a Dual-Axis Rotation Inertial Navigation System.
In the dual-axis rotation inertial navigation system (INS), besides the gyro error, accelerometer error, rolling misalignment angle error, and the gimbal angle error, the shaft swing angle and the axis non-orthogonal angle also affect the attitude accuracy. Through the analysis of the structure, we can see that the shaft swing angle and axis non-orthogonal angle will produce coning errors which cause the fluctuation of the attitude. According to the analysis of the rotation vector, it can be seen that the coning error will generate additional drift velocity along the rotating shaft, which can reduce the navigation precision of the system. In this paper, based on the establishment of the modulation average frame, the vector projection is carried out, and then the attitude conversion matrix and the attitude error matrix mainly including the shaft swing angle and axis non-orthogonal are obtained. Because the attitude angles are given under the static condition, the shaft swing angle and the axis non-orthogonal angle are estimated by the static Kalman filter (KF). This kind of KF method has been widely recognized as the standard optimal estimation tool for estimating the parameters such as coning angles (α₁ , α₂), initial phase angles (ϕ₁,ϕ₂), and the non-perpendicular angle (η). In order to carry out the system level verification, a dual axis rotation INS is designed. Through simulation and experiments, the results show that the amplitudes of the attitude angles' variation are reduced by about 20%-30% when the shaft rotates. The attitude error equation is reasonably simplified and the calibration method is accurate enough. The attitude accuracy is further improved.


[Effect of minor bioactive food substances – rutin and hesperidin in their separate and combined alimentary arrives on the immune system of rats and the activity of nuclear factor NF-kB liver cells].
The effect of rutin and hesperidin in their separate and combined admission to the immune system and the activity of nuclear factor NF-kB of rat liver cells has been investigated. Wistar male rats with an initial body weight of 224–225 g were divided into 4 groups of 6 rats in each. The rats of the 1st group (control) received a complete semi-synthetic diet, rats in group 2 – the same diet supplemented with rutine (400 mg/kg b.w.); the rats of group 3 – with the addition of hesperidin (400 mg/kg bw); group 4 – with the addition of rutin and hesperidin (400 mg/kg b.w. each) for 14 days. Animals received feed in free access mode in an amount of 25–30 g per rat per day, that corresponded to 15 g of dry formula. Animals received water also in free access. It has been found that rutin and hesperidin, included in the diet of rats both alone (groups 2 and 3) and together (group 4), have immunomodulatory impact which is a reduce of lymphocyte relative content [1st gr. – 70.55±1.58%, 2nd gr. – 63.62±2.85%, 3rd gr. – 62.03±3.16% (p1–3<0.05), 4th gr. – 65.75±1.08% (p1–4<0.05)] and an increase of percentage of neutrophil leukocytes [1st gr – 19.98±0.97%, 2nd gr. – 25.35±3.14%, 3rd gr. – 28.27±3.30% (p1–3<0.05), 4th gr. – 24.15±1.52% (p1–4<0.05)] and NK-cells in the peripheral blood [1st gr. – 3.29±0.45%, 2nd gr. – 6.91±0.70% (p1–2<0.05), 3rd gr. – 5.88±0.79% (p1–3<0.05), 4th gr. – 4.64±0.32% (p1–4<0.05)], that can be considered as a shift in the direction of innate immunity factors. In addition, the combined effect of high doses of rutin and hesperidin led to a change in erythrocyte parameters: an increase in the average volume of red blood cells [1st gr. – 56.00±1.06 fl, 2nd gr. – 56.67±0.42 fl, 3rd gr. – 58.50± 0.99 fl, 4th gr. – 59.50±0.99 fl (p1–4<0.05)], and the average content of hemoglobin [1st gr. – 18.97±0.45 pg, 2nd gr. – 19.10±0.19 pg, 3rd gr. – 19.73± 0.32 pg, 4th gr. – 20.08±0.33 pg (p1–4=0.07)], as well as increase in the level of TGF-β1 in peripheral blood [1st gr. – 15.55±2.13 ng/ml, 2nd gr. – 14.81± 2.36 ng/ml, 3rd gr. – 17.02±2.53 ng/ml, 4th gr. – 22.14±2.29 ng/ml (p1–4<0.05)] and the expression of nuclear factor NF-kB in the liver cells [1st gr. – 16.10± 0.60 ng/ml; 2nd gr. – 15.14±2.28 ng/ml; 3rd gr. – 15.85±2.09 ng/ml; 4th gr. – 20.49±1.68 ng/ml (p1–4<0.05)].


[Association between sphericity, ventricular function and size of the infarction in rats].
The left ventricular (LV) sphericity is a factor associated with ventricular dysfunction, but it is not well-characterized in the experimental infarction model in rats. To analyze the association between the sphericity index (SI), the ventricular function and the infarcted area in an experimental rat model. Six months after the infarction (AMI, n=33) or simulated surgery (SHAM, n=18), the animals were submitted to an echocardiogram. The SI was obtained through the ratio between the diastolic areas at the LV long axis and the short axis. The AMI group presented the lowest index of sphericity (1.32 x 0.23 vs 1.57 x 0.33; p=0.002), systolic function and relative thickness (0.13 x 0.003 vs 0.18 x 0.04; p<0.001) and the highest index of parietal stress (1.27 x 0.33 vs 0.88 x 0.25; p<0.001). There was a significant correlation between the infarct size and sphericity (p=0.046). At the linear regression analysis, the infarct size (p=0.014), but not the sphericity (p=0.683) and the parietal stress (p=0.176), was the predictive factor of the systolic function. Eccentric remodeling (p=0.011), but not sphericity (p=0.183) or the infarct size (p=0.101), was a predictive factor of parietal stress. Additionally, the infarct size (p=0.046), but not the eccentric remodeling (0.705), was a predictive factor of sphericity. The infarct size (p=0.015) and the parietal stress (p=0.011), but not the sphericity (p=0.705), were the predictors of eccentric remodeling. The sphericity is associated, but it is not a determinant factor of parietal stress, of eccentric remodeling and ventricular systolic function in an experimental infarction model in rats.


Long-term care of pediatric renal transplant patients: from bench to bedside.
In this review, we discuss current and future issues in the management of pediatric renal transplant recipients, including the optimization of long-term graft function and the minimization of complications caused by immunosuppression. Long-term management involves not only the monitoring of graft function but also the identification of patients at risk for the development of complications. The identification of patients with immunoreactive or immunoregulatory responses can be performed molecular monitoring of the immune response. Also, the use of frequent surveillance kidney biopsies, surrogate markers of chronic rejection, and glomerular filtration rate will be a part of future management. Identifying high-risk patients enables the physician to optimize immunosuppression to limit acute rejection. Short-and long-term management of pediatric transplant patients also includes adequate monitoring of growth and the monitoring for post-transplant lymphoproliferative disease. Ongoing clinical trials are underway that focus on these novel approaches in caring for pediatric transplant recipients.


Management of subluxated capsular bag-fixated intraocular lenses using a capsular anchor.
We describe the use of the capsular anchor (AssiAnchor) to manage a subluxated intraocular lens (IOL) in the capsular bag. The anchor comprises 2 prongs that hold the anterior lens capsule and a central rod that is sutured to the scleral wall, enabling centration of the IOL-capsular bag complex. Six pseudophakic patients presenting with subluxated posterior chamber IOLs in the capsular bag were operated on using the device. The anchor was used successfully in all cases, although in 2 cases only 1 prong was placed under the capsulorhexis edge. In 1 eye, 2 anchors were used 1 month apart following repeated traumatic zonular injury. The capsular bag holding the IOL remained centered and stable throughout the follow-up period. The anchoring device, which was originally designed to preserve the lens capsule and stabilize subluxated crystalline lenses, can also be used to treat subluxation of a capsular bag-fixated IOL. Dr. Assia is the inventor of the AssiAnchor, has a licensed patent of the anchor, and is consultant to Hanita Lenses. Dr. Lapid-Gortzak is a consultant to and speaker for Alcon Surgical, Inc., Hanita Lenses, Orca Surgical, and Sanoculis Ltd.; a speaker for Santen; and a consultant to Icon. Drs. Ton and Naftali have no financial or proprietary interest in any material or method mentioned.


Circadian rhythm variations in the adult rat induced by low and high protein diets administered at various stages of development.
Hypothalamic tryptophan (TP), serotonin (5HT) and 5-hydroxy-indoleacetic acid (5HIAA), serum tryptophan (free and total), and circulating corticosterone and aldosterone were measured over 24 hr in 60-day-old male rats fed high (60%) and low (6%) protein diets. The diets were administered during specific stages of rat growth: fetal phase and suckling (when the process of mitosis is most rapid), post-lactation, and from the fetal phase through to the adult stage. In all groups, there was a direct correlation between protein intake and hypothalamic and serum TP levels. Remarkable decreases in the amplitude and mesor of the aldosterone rhythm were observed when the hyperprotein diet was administered in the fetal and suckling phases, or from the fetal phase until the adult stages. The rhythm of the serotonin system was significantly altered by a low protein diet: decreased 5-HT amplitude (low protein diet in the fetal stage and suckling), decreased 5-HT amplitude and deranged 5-HIAA rhythm (low protein in the post-lactation period), deranged 5-HIAA rhythm (low protein diet throughout the experiment). Lastly, there was a remarkable increase in the corticosterone mesor in rats administered a low protein diet after weaning and in rats constantly given this diet. Growth was greatly inhibited in these two groups of animals, therefore, it cannot be excluded that the high levels of circulating hormone resulted from their debilitated condition.


[Interdisciplinary efforts in public health: an area under construction].
This article, based on a literature review, aims to discuss the interdisciplinarity on Public Health (its needs, barriers for implementation and possibilities). Therefore, initially, the authors present a discuss on conceptual and historical terms, that explains the subject. In this part, it is discussed the process of inclusion and exclusion of interdisciplinarity on Public Health, as well as the different dimension of it. Subsequently it is related an interdisciplinary experience on Public Health. Finally, the authors concluded that the interdisciplinary is extremely necessary to Public Health due to the complexity of its object of study.


Relationships between tumor microenvironment and clinicopathological parameters in meningioma.
To evaluate the relationship between the subtype of cells/cellular constituents (the density of T lymphocyte subsets, B lymphocyte, macrophages, and FOXP3 positive cells in 93 patients with meningioma, WHO grades I and II) in the tumor microenvironment and clinicopathological parameters (gender, age, tumor location, size, recurrence and pathological type) of meningioma. Immunohistochemical demonstrations of CD20 and CD4 lymphocytes, CD68, and FOXP3 expression were performed. In order to assess the densities of CD4, CD20, CD68 and FOXP3 positive cells in 93 meningioma patients, the results were derived from independent reviews by two pathologists. Chi-square test was used for independent samples. There were no relationships between the CD4(+), CD68(+) cell subsets and patients' age, sex, tumor size, grade and the recurrence of tumor. However, patients with recurrence had a significantly higher density of CD20(+) B cells compared to patients with no recurrence (P = 0.003). For the Foxp3(+) cell subset, results showed us that more female patients had high density of Foxp3(+) cells compared with male patients, while the opposite results were observed in the low density group (P = 0.009). Furthermore, the density of Foxp3(+) cells was significantly correlated with the tumor size (P = 0.004) and the pathological types (P = 0.004). Results in this study demonstrate that higher CD20(+) B cell density in the tumor is associated with lower tumor recurrence and the density of Foxp3(+) cells is significantly correlated with the patients' sex, tumor size and the pathological types. The results also suggest that understanding of the cellular constituents of tumors and the tumor microenvironment may help investigate the tumor pathogenesis and immunotherapies in meningioma.


Pathology, danger, and power: women's and physicians' views of pregnancy and childbirth in Weimar Germany.
This article shows how the very real risks of reproduction were culturally constituted by expectant mothers and physicians in Germany following the first World War. Both saw a strong potential for maternal and fetal death and injury. However, expectant mothers and physicians differed in the extent to which they regarded risk as contingent or inherent. Most physicians believed that pregnancy and childbirth first became dangerous in conjunction with a wide variety of other ailments, including complications specific to pregnancy. This view of pathology held a promise of prevention or at least treatment. Physicians further participated in inventing new, modern understandings of pathology, which also encompassed factory work as a reproductive hazard. Expectant mothers, in contrast, viewed pregnancy and childbirth as inherently dangerous based on their experiences and those of the women they knew, which not infrequently involved illness, disability, or close encounters with death. Many of them also held the ancient idea that a woman could harm the foetus with her imagination. As a result, though women varied in their beliefs (influenced by class, education, region, and religion), as a group they feared childbirth. Paradoxically, competition among lay and medical ideas gave women new options while also gradually enhancing medical authority.


Populations of western North American monkeyflowers accrue niche breadth primarily via genotypic divergence in environmental optima.
Niche breadth, the range of environments that individuals, populations, and species can tolerate, is a fundamental ecological and evolutionary property, yet few studies have examined how niche breadth is partitioned across biological scales. We use a published dataset of thermal performance for a single population from each of 10 closely related species of western North American monkeyflowers (genus Mimulus) to investigate whether populations achieve broad thermal niches through general purpose genotypes, specialized genotypes with divergent environmental optima, and/or variation among genotypes in the degree of generalization. We found the strongest relative support for the hypothesis that populations with greater genetic variation for thermal optimum had broader thermal niches, and for every unit increase in among-family variance in thermal optimum, population-level thermal breadth increased by 0.508°C. While the niche breadth of a single genotype represented up to 86% of population-level niche breadth, genotype-level niche breadth had a weaker positive effect on population-level breadth, with every 1°C increase in genotypic thermal breadth resulting in a 0.062°C increase in population breadth. Genetic variation for thermal breadth was not predictive of population-level thermal breadth. These findings suggest that populations of Mimulus species have achieved broad thermal niches primarily through genotypes with divergent thermal optima and to a lesser extent via general-purpose genotypes. Future work examining additional biological hierarchies would provide a more comprehensive understanding of how niche breadth partitioning impacts the vulnerabilities of individuals, populations, and species to environmental change.


A case of atypical progressive outer retinal necrosis after highly active antiretroviral therapy.
This is a report of an atypical case of progressive outer retinal necrosis (PORN) and the effect of highly active antiretroviral therapy (HAART) on the clinical course of viral retinitis in an acquired immunodeficiency syndrome (AIDS) patient. A 22-year-old male patient infected with human immunodeficiency virus (HIV) presented with unilaterally reduced visual acuity and a dense cataract. After cataract extraction, retinal lesions involving the peripheral and macular areas were found with perivascular sparing and the mud-cracked, characteristic appearance of PORN. He was diagnosed as having PORN based on clinical features and was given combined antiviral treatment. With concurrent HAART, the retinal lesions regressed, with the regression being accelerated by further treatment with intravenous acyclovir and ganciclovir. This case suggests that HAART may change the clinical course of PORN in AIDS patients by improving host immunity. PORN should be included in the differential diagnosis of acute unilateral cataract in AIDS patients.


Chronic daily ethanol and withdrawal: 3. Forebrain pro-opiomelanocortin gene expression and implications for dependence, relapse, and deprivation effect.
Although forebrain pro-opiomelanocortin (POMC)-producing neurons seem to mediate or modulate many responses to ethanol consumption, changes in activity of this opiomelanocortinergic system in response to chronic ethanol consumption, withdrawal, and subsequent abstinence remain unresolved. We investigated the effects of chronic daily ethanol consumption, withdrawal, and subsequent abstinence on adult male Sprague-Dawley rat forebrain opiomelanocortinergic activity as reflected by changes in hypothalamic POMC messenger RNA (mRNA) content by using a well characterized liquid diet model that we have previously demonstrated to accurately simulate not only daily oral ethanol consumption quantity and pattern, but also both neuroendocrine and behavioral changes characteristic of actively drinking and subsequently abstinent alcoholics. After 7 weeks of daily ethanol consumption at night and withdrawal during the day, evening mediobasal hypothalamus POMC mRNA concentrations were suppressed versus both ad libitum-fed and pair-fed controls. Morning POMC mRNA concentrations were also suppressed versus ad libitum-fed controls and tended to be decreased versus pair-fed controls. Three weeks after gradual removal of ethanol from the diet, mediobasal hypothalamus POMC mRNA concentrations were increased relative to ad libitum-fed and pair-fed controls. Plasma concentrations of corticosterone, testosterone, and leptin were also altered by the daily ethanol/withdrawal treatment and by subsequent abstinence. Because each of these hormones has been demonstrated to modify forebrain POMC gene expression under some conditions, the overall changes in forebrain opiomelanocortinergic regulation in response to chronic daily ethanol/withdrawal and subsequent abstinence probably reflect, at least in part, regulation by multiple endocrine mechanisms, together with responses to stress, development of tolerance during chronic daily ethanol consumption, and rebound of function after termination of this consumption. Overall, the demonstrated changes in forebrain POMC gene expression are consistent with significant roles for forebrain opiomelanocortinergic regulation in mediating alcohol dependence, propensity to relapse, and the alcohol deprivation effect.


Infant hearing screening.
Infant hearing screening has progressed markedly over the past year. Although uncommon in the past, now almost 95% of infants in the United States are screened for hearing loss. Recent literature has expanded on several important components of hearing screening. A major revision of the American Academy of Pediatrics policy statement on infant hearing screening was published. This statement identifies a number of important principles and guidelines for infant hearing screening. Improvements in diagnostic techniques have been described including the need for auditory brainstem response screening in premature infants. Risk factors for congenital-hearing loss have been updated. An etiologic diagnosis for infants with hearing loss is increasingly possible with advances in genetics and molecular diagnosis. Finally, several articles provide further information on optimizing follow-up and diagnostic testing as well as early intervention. Despite the challenges still present in infant hearing screening, screening continues to identify infants at a very young age, and improve the early communication skills of infants with hearing loss through early diagnosis and early intervention.


A brain-tumor neural circuit controls breast cancer progression in mice.
Tumor burden, considered a common chronic stressor, can cause widespread anxiety. Evidence suggests that cancer-induced anxiety can promote tumor progression, but the underlying neural mechanism remains unclear. Here, we used neuroscience and cancer tools to investigate how the brain contributes to tumor progression via nerve-tumor crosstalk in mice model of breast cancer. We showed that the tumor-bearing mice exhibited significant anxiety-like behaviors and that corticotropin-releasing hormone (CRH) neurons in the central medial amygdala (CeM) were activated. Moreover, newly formed sympathetic nerves were detected in tumors, which established a polysynaptically connected with the brain. Pharmacogenetic or optogenetic inhibition of CeMCRH neurons and CeMCRH→LPGi circuit significantly alleviated anxiety-like behaviors and slowed tumor growth. Conversely, artificial activation of CeMCRH neurons and CeMCRH→LPGi circuit increased anxiety and tumor growth. Importantly, alprazolam, an anti-anxiety drug, was found to be a promising intervention for slowing tumor progression. Furthermore, we showed that manipulation of CeMCRH→LPGi circuit directly regulates the activity of the intratumoral sympathetic nerves and peripheral nerve-derived norepinephrine, then affecting tumor progression by modulating the antitumor immunity. Together, these findings reveal a brain-tumor neural circuit contributing to breast cancer progression and provide new therapeutic insights for breast cancer.


Postinfectious cerebellitis: clinical significance of Tc-99m HMPAO brain SPECT compared with MRI.
We compared Tc-99m HMPAO brain SPECT and MRI with the clinical findings in a case of a 14-year-old boy with post-infectious cerebellitis. Tc-99m HMPAO brain SPECT scan showed marked reduction in perfusion to the left anterior lobe, left lateral hemisphere, and left posterior lobe of the cerebellum. The MRI showed a limited region of abnormal signal in the mid posterior left cerebellar hemisphere. Since the patient's overall neurological deficit was characterized by ataxia, nystagmus, head and body titubation, dysmetria, dysdiadochokinesia, loss of normal check, and rebound testing attributed to the involvement of the anterior, lateral, and posterior lobes of the left cerebellar hemisphere, the Tc-99m HMPAO brain SPECT finding correlated better with the extent of functional neurological deficits. Tc-99m HMPAO brain SPECT may better assess cerebellar signs and symptoms in cerebellitis since the blood flow abnormalities correlated with dysfunction of the cerebellum, whereas the MRI abnormalities did not encompass the scope of the neurological deficits.


Trypanosoma brucei and Trypanosoma cruzi DNA Mismatch Repair Proteins Act Differently in the Response to DNA Damage Caused by Oxidative Stress.
MSH2, associated with MSH3 or MSH6, is a central component of the eukaryotic DNA Mismatch Repair (MMR) pathway responsible for the recognition and correction of base mismatches that occur during DNA replication and recombination. Previous studies have shown that MSH2 plays an additional DNA repair role in response to oxidative damage in Trypanosoma cruzi and Trypanosoma brucei. By performing co-immunoprecipitation followed by mass spectrometry with parasites expressing tagged proteins, we confirmed that the parasites' MSH2 forms complexes with MSH3 and MSH6. To investigate the involvement of these two other MMR components in the oxidative stress response, we generated knockout mutants of MSH6 and MSH3 in T. brucei bloodstream forms and MSH6 mutants in T. cruzi epimastigotes. Differently from the phenotype observed with T. cruzi MSH2 knockout epimastigotes, loss of one or two alleles of T. cruzi msh6 resulted in increased susceptibility to H2O2 exposure, besides impaired MMR. In contrast, T. brucei msh6 or msh3 null mutants displayed increased tolerance to MNNG treatment, indicating that MMR is affected, but no difference in the response to H2O2 treatment when compared to wild type cells. Taken together, our results suggest that, while T. cruzi MSH6 and MSH2 are involved with the oxidative stress response in addition to their role as components of the MMR, the DNA repair pathway that deals with oxidative stress damage operates differently in T. brucei.


[Successful treatment of panarteritis nodosa with low-dose methotrexate therapy].
We report on a 45-year-old male patient who presented a classic polyarteritis nodosa (PAN). The clinical course extended over 7 years. In spite of 2 years immunosuppressive therapy with azathioprine and methylprednisolone the course was progressive. Low-dose methotrexate therapy was the only treatment that controlled the disease, leading to rapid clinical and histopathological remission. In low concentrations methotrexate acts as an IL-1 inhibitor, and it obviously suppresses the pathogenetic mechanism of PAN.


FOXP3 and CXCR4-positive regulatory T cells in the tumor stroma as indicators of tumor immunity in the conjunctival squamous cell carcinoma microenvironment.
Conjunctival squamous cell carcinoma (SCC) is the most common ocular surface neoplasia. The purpose of this retrospective study was to examine the role of regulatory T cell (Treg) activity in tumor immunity and investigate the tumor microenvironment as a new treatment focus in conjunctival SCC. Cancer progression gene array and immunohistochemical analyses of FOXP3 as a Treg marker, CD8 as a tumor-infiltrating lymphocyte marker, and CXCR4 expression on activated Tregs were conducted in a series of 31 conjunctival SCC cases. The objective was to investigate the immunoreactive response in tumor cells and stromal cells in the cancer microenvironment. The stroma ratio in tumor cells was investigated by monitoring α-smooth muscle actine (SMA) expression between carcinoma in situ (Tis) and advanced carcinoma (Tadv) (P<0.01). No significant change in PD-L1 expression was observed in this study (P = 0.15). Staining patterns of FOXP3, CD8, and CXCR4 were examined separately between tumor cells and stromal cells in SCC tumors. Differences in staining of FOXP3 in Tregs and CD8 in tumor-infiltrating lymphocytes in tumor stroma in the Tis group were observed compared with the Tadv group (each P<0.01). In addition, double immunostaining of CXCR4/FOXP3 was correlated with progression-free survival (P = 0.049). Double immunostaining of CXCR4/FOXP3 correlated with American Joint Committee on Cancer T-stage, independent of age or Ki67 index (P<0.01). Our results show that FOXP3 and the CXCR4/FOXP3 axis are important pathologic and prognostic factors of ocular surface neoplasia, including SCC. The tumor microenvironment of conjunctival SCC should be considered in the future development of treatment options.


Increased antioxidant efficacy of tocopherols by surfactant solubilization in oil-in-water emulsions.
The physical location of antioxidants in oil-in-water emulsions can have significant influence on their free radical scavenging activity and ability to inhibit lipid oxidation. We aimed to determine the effect of the surfactant concentration on the partitioning behavior of tocopherols (α, γ, and δ) in oil-in-water emulsions. Tween 20 (0.1, 0.5, and 1%) increased the partitioning of the tocopherols into the aqueous phase via the formation of Tween 20-tocopherol comicelles. Partitioning behavior of antioxidants was dependent upon the number of methyl groups and, thus, polarity of the tocopherols. δ-Tocopherol (one methyl group) exhibited the most partitioning into the aqueous phase, while α-tocopherol (three methyl groups) had the lowest partitioning. Lipid oxidation studies showed that the antioxidant activity of δ- and α-tocopherols was enhanced by adding Tween 20 to oil-in-water emulsions. This work suggests that surfactant micelles could increase the antioxidant activity of tocopherols by changing their physical location.


Novel bacterial consortia isolated from plastic garbage processing areas demonstrated enhanced degradation for low density polyethylene.
This study aimed to formulate novel microbial consortia isolated from plastic garbage processing areas and thereby devise an eco-friendly approach for enhanced degradation of low-density polyethylene (LDPE). The LDPE degrading bacteria were screened and microbiologically characterized. The best isolates were formulated as bacterial consortia, and degradation efficiency was compared with the consortia formulated using known isolates obtained from the Microbial Culture Collection Centre (MTCC). The degradation products were analyzed by FTIR, GC-FID, tensile strength, and SEM. The bacterial consortia were characterized by 16S ribosomal DNA (rDNA) sequencing. The formulated bacterial consortia demonstrated 81 ± 4 and 38 ± 3 % of weight reduction for LDPE strips and LDPE pellets, respectively, over a period of 120 days. However, the consortia formulated by MTCC strains demonstrated 49 ± 4 and 20 ± 2 % of weight reduction for LDPE strips and pellets, respectively, for the same period. Furthermore, the three isolates in its individual application exhibited 70 ± 4, 68 ± 4, and 64 ± 4 % weight reduction for LDPE strips and 21 ± 2, 28 ± 2, 24 ± 2 % weight reduction for LDPE pellets over a period of 120 days (p < 0.05). The end product analysis showed structural changes and formation of bacterial film on degraded LDPE strips. The 16S rDNA characterization of bacterial consortia revealed that these organisms were novel strains and designated as Enterobacter sp. bengaluru-btdsce01, Enterobacter sp. bengaluru-btdsce02, and Pantoea sp. bengaluru-btdsce03. The current study thus suggests that industrial scale-up of these microbial consortia probably provides better insights for waste management of LDPE and similar types of plastic garbage.


Restoration of primary canines with porcelain laminate veneers: a clinical report.
This article describes treatment of a young adult patient with porcelain laminate veneers for restoring unaesthetic maxillary anterior teeth with two retained primary canines. The patient had experienced an approximately two-year orthodontic treatment and had received both fixed and removable retainers for the upper arch. The patient could not afford implant supported restorations for his missing premolar teeth and was not pleased by the appearance of his smile. Using porcelain laminate veneers is a proper treatment option that could be taken into consideration in these situations.


Temporal Evaluation of Polybrominated Diphenyl Ether (PBDE) Serum Levels in Middle-Aged and Older California Women, 2011-2015.
In response to health concerns and widespread human exposures, two widely used commercial formulations of polybrominated diphenyl ethers (PBDEs) were banned in the United States in 2005. Initial biomonitoring data have provided early indications of reduced human exposures since these bans took effect. Our objective was to evaluate temporal trends in PBDE serum levels among a population of older California women during a four-year period, beginning approximately five years after these formulations were banned. Automated solid phase extraction and gas chromatography/high resolution mass spectrometry were used to measure PBDE levels in blood collected during 2011-2015 among 1253 women (ages 40-94) participating in the California Teachers Study. Only congeners with detection frequencies (DF) ≥ 75% were included in the present analysis: BDE-47 (DF = 88%); BDE-100 (DF = 78%); and BDE-153 (DF = 80%). Results from age- and race/ethnicity-adjusted linear regression analyses indicated modest, but statistically significant, average annual percent increases in the serum concentrations of all three PBDEs over the four-year study period. While not without limitations, these results, in the context of other biomonitoring data, suggest that earlier reported declines in PBDE levels may have plateaued and may now be starting to increase. Further biomonitoring to ascertain current trends and determinants of population exposures is warranted.


Extracts of the Wild Potato Species Solanum chacoense on Breast Cancer Cells: Biochemical Characterization, In Vitro Selective Cytotoxicity and Molecular Effects.
Solanum chacoense (wild potato) is intensively used in breeding, its biochemical profile and putative human health-related traits being transferred into potato cultivars aimed for consumption. The goal of this study was to evaluate the biochemical profile and the anti-tumor potential of methanolic extracts obtained from S. chacoense leaves and tubers against three breast cancer cell lines in comparison to healthy endothelial cells (HUVEC). The biochemical profile of the extracts was determined by HPLC-PDA/-ESI+-MS and ITEX/GC-MS, the selective cytotoxicity by MTT assay whereas RT-qPCR was used to evaluate the expression of proliferation- and apoptosis-related genes. Both extracts proved to be rich in phenolic acids and volatile compounds, the leaf extract also containing glycoalkaloids. Both extracts proved to be cytotoxic for breast cancer cell lines, with IC50 values varying between 132.9 and 390.7 µg/ml. Both extracts had selective cytotoxicity against MCF7 cell line in comparison to HUVECs (selectivity coefficients >2.3). The treatment with the extracts induced overexpression of the pro-apoptotic gene BAX¸ down-regulation of the anti-apoptotic gene BCL-2 and the pro-proliferation genes NFkB, CCND1, and STAT3. Thus S. chacoense extracts proved to be rich in compounds with anticancer proprieties and are capable of inducing selective cytotoxicity on MCF7 cell line.


Synthetic intron improves transduction efficiency of trans-splicing adeno-associated viral vectors.
Trans-splicing adeno-associated viral (AAV) vectors hold great promise in many gene therapy applications. We have shown that rational selection of the gene-splitting site in a therapeutic target gene can lead to extremely efficient trans-splicing vectors [Lai, Y., Yue, Y., Liu, M., Ghosh, A., Engelhardt, J.F., Chamberlain, J.S., and Duan, D. (2005). Nat. Biotechnol. 23, 1435-1439]. Our original strategy requires the screening of endogenous introns that are capable of overcoming the mRNA accumulation barrier. To further develop transsplicing vectors, we have tested whether the use of a generic synthetic intron can bypass the labor-intensive intron-screening process. Two previously characterized exon/intron/exon junctions (60/60/61 and 63/63/64, respectively) in the 6 kb minidystrophin gene were used as templates to represent highly efficient (60/60/61) and relatively poor (63/63/64) gene-splitting sites. We compared RNA production from the reconstituted viral genome and transduction efficiency of the trans-splicing vectors in dystrophin-null mdx mouse skeletal muscle. Our results suggest that a synthetic intron can successfully overcome the mRNA accumulation barrier at the exon 63/64 junction. Furthermore, when the gene was split at the exon 63/64 junction, the synthetic intronbased vectors performed better than the endogenous intron-based vectors. When the gene was split at the exon 60/61 junction, we observed only nominal improvement in mRNA production. Nevertheless, vectors based on the exon 60/61 junction remain the best set in transduction efficiency. Taken together, our results suggest that optimizing intron sequence may boost the transduction efficiency of trans-splicing AAV vectors.


Requirement of matrix metalloproteinase-9 for the transformation of human mammary epithelial cells by Stat3-C.
Persistently activated Stat3 is found in many different cancers, including approximately 60% of breast tumors. Here, we demonstrate that a constitutively activated Stat3 transforms immortalized human mammary epithelial cells and that this oncogenic event requires the activity of matrix metalloproteinase-9 (MMP-9). By immunohistochemical analysis, we observe a positive correlation between strong MMP-9 expression and tyrosine phosphorylated Stat3 in primary breast cancer specimens. These results demonstrate a relationship between activated Stat3 and MMP-9 in breast oncogenesis.


GREM1 is expressed in the cancer-associated myofibroblasts of basal cell carcinomas.
Cancer-associated fibroblasts (CAFs) play important roles in cancer progression through their complex interactions with cancer cells. The secreted bone morphogenetic protein antagonist, gremlin1 (GREM1) is expressed by the CAFs of basal cell carcinomas (BCCs), and promotes the growth of cancer cells. In this study, we investigated the expression of GREM1 mRNAs in various benign and malignant skin tumors, including various BCC subtypes. Analysis by RNA in situ hybridization (ISH) revealed that fibroblasts in the scar tissue expressed GREM1 and α-smooth muscle actin (α-SMA), whereas resident fibroblasts in the dermis of the normal skin did not express GREM1. Real-time polymerase chain reaction analysis showed significantly higher GREM1 expression in skin cancers and pilomatricomas (PMCs) than in other benign skin tumors. Tissue microarrays analyzed by RNA ISH for GREM1 expression also demonstrated that 23% of BCCs, 42% of squamous cell carcinomas, 20% of melanomas, and 90% of PMCs were positive for GREM1 expression, whereas trichoepitheliomas, eccrine poromas, hidradenomas, and spiradenomas were negative for GREM1 expression. Most BCCs that were GREM1 expression positive were of desmoplastic or mixed subtypes, and GREM1 expression was localized to activated myofibroblasts at the tumoral-stromal interface. Interestingly, most PMCs harbored GREM1-expressing fibroblasts, probably because of the inflammatory responses caused by foreign body reactions to keratin. Additionally, in BCCs, stromal GREM1 expression had a strong correlation with CD10 expression. In conclusion, GREM1 is frequently expressed by myofibroblasts in scars or in the stroma of basal cell carcinomas, suggesting that GREM1 expression can be a marker for activated myofibroblasts in the cancer stroma or in scar tissue.


Polymorphism of the HLA-DP beta region detected by Southern blot hybridization.
A panel of homozygous cell lines, previously typed by primed lymphocyte test for their DPw specificity, have been studied by restriction fragment length polymorphism analysis, using a DP beta-specific probe. Highly stringent hybridization and washing conditions were used to prevent cross-hybridization with DR- and DQ-specific fragments. Three out of six enzymes employed allowed us to distinguish clustered or single DPw specificities, and by MspI digestion it was possible to detect different patterns within a single specificity such as DPw4. Some of the cell lines have been further studied with synthetic oligonucleotides derived from the polymorphic regions of the second exon of DP beta 1 gene, and, in general, a correlation with the primed lymphocyte test--defined specificities and restriction fragment length polymorphism was found. These data suggest a more extended complexity of the DP region, in addition to that defined as the DPw1-DPw6 segregant series.


Pain management during labour at the El-Emarati Hospital: a clinical audit.
The aim of this study was to assess the practice of pain management for women in labour at El-Emarati Hospital in Rafah, Gaza Strip, and to compare this management with the guidelines by the National Institute for Health and Care Excellence (NICE). A data collection sheet was prepared and completed for each patient admitted to the delivery room. Patients were enrolled consecutively between Feb 1 and April 1, 2016. Data collection sheets, based on NICE guidelines, were completed by the audit team (TAH, MSF, and RYA). Verbal consent was obtained from all participants. 50 patients were included in the audit. 22 patients were primigravidae, and 28 patients were multigravidae. At least one or more non-pharmacological pain management method was used for each patient. 50 (100%) patients were managed by breathing and relaxing techniques. 35 (70%) patients also received abdominal massage. 20 (40%) patients had their pain managed pharmacologically. None of the patients was informed beforehand about possible side-effects of the drugs used, and none of the patients was offered or received regional anaesthesia for pain management during labour. We found a substantial shortfall in the management of pain during labour, especially as many options and techniques are not used or offered to patients. Another weakness was the limited discussion and lack of choice available to patients with respect to the use of pain relief. Pain is a highly subjective experience and varies greatly between patients, so preferences for use of pain relief will differ between individuals, and this should be reflected in practice. None.


Patient satisfaction in military medicine: status and an empirical test of a model.
The Department of Defense (DoD) is concerned about how well military medical treatment facilities in the military health system perform. Patient expectations, attitudes, and health care use have been examined in numerous studies; the results are fairly consistent. Eligible beneficiaries report moderate satisfaction with the health care received. In 1994-2001, annual DoD and monthly ambulatory patient surveys were conducted in military medical treatment facilities. The DoD surveys document how patients perceive the care provided. The obvious research concerns are: requirements for conducting surveys; who should be surveyed: eligible beneficiaries or actual users; when; where; representative sample; how often to conduct assessment; data collection methods; analytic schemes; overall trends; predictors of satisfaction; use of results; and timeliness of findings. This study examines these issues and analyzes raw data from selected annual DoD and monthly ambulatory surveys. The overall level of perceived satisfaction has been "good" over the years surveys were used. The model demonstrated the use of examining demographic and attitudinal components of patient satisfaction in military medical facilities.


Idiopathic hepatic arterio-portal fistula: report of one case.
Hepatic arterio-portal fistula is a rare cause of portal hypertension in children; it is an abnormal communication of hepatic artery and portal venous system, the most common causes being trauma or malignancy. There were only 11 cases reported in English literature and were not ever reported in Taiwan. We report a 9-year old boy with idiopathic hepatic arterio-portal fistula presented as intractable hematemesis due to esophageal and gastric varices. He had received sclerotherapy twice, and Sugiura operation (resection of the lower part of esophagus, devasculization of the stomach and splenectomy). Idiopathic hepatic arterio-portal fistula was found in angiography examination and the esophageal and gastric varices disappeared after transarterial embolization (TAE). We conclude that angiography is the golden diagnostic method for portal hypertension when the etiology is hepatic arterio-portal fistula and TAE will provide immediately therapy.


SNPs in Mammary Gland Epithelial Cells Unraveling Potential Difference in Milk Production Between Jersey and Kashmiri Cattle Using RNA Sequencing.
Deep RNA sequencing experiment was employed to detect putative single nucleotide polymorphisms (SNP) in mammary epithelial cells between two diverse cattle breeds (Jersey and Kashmiri) to understand the variations in the coding regions that reflect differences in milk production traits. The low milk-producing Kashmiri cattle are being replaced by crossbreeding practices with Jersey cattle with the aim of improving milk production. However, crossbred animals are prone to infections and various other diseases resulting in unsustainable milk production. In this study, we tend to identify high-impact SNPs from Jersey and Kashmiri cows (utilizing RNA-Seq data) to delineate key pathways mediating milk production traits in both breeds. A total of 607 (442 SNPs and 169 INDELs) and 684 (464 SNPs and 220 INDELs) high-impact variants were found specific to Jersey and Kashmir cattle, respectively. Based on our results, we conclude that in Jersey cattle, genes with high-impact SNPs were enriched in nucleotide excision repair pathway, ABC transporter, and metabolic pathways like glycerolipid metabolism, pyrimidine metabolism, and amino acid synthesis (glycine, serine, and threonine). Whereas, in Kashmiri cattle, the most enriched pathways include endocytosis pathway, innate immunity pathway, antigen processing pathway, insulin resistance pathway, and signaling pathways like TGF beta and AMPK which could be a possible defense mechanism against mammary gland infections. A varied set of SNPs in both breeds, suggests a clear differentiation at the genomic level; further analysis of high-impact SNPs are required to delineate their effect on these pathways.