Approval summary for imatinib mesylate capsules in the treatment of chronic myelogenous leukemia.
Chronic myelogenous leukemia (CML) results from the breakpoint cluster region-Abl fusion gene product, a tyrosine kinase involved in cell division and apoptosis. Imatinib, an orally administered inhibitor of the breakpoint cluster region-Abl tyrosine kinase, is capable of blocking proliferation and inducing apoptosis in CML cell lines. In this report, we describe the preclinical profile of imatinib and the data submitted in the New Drug Application that led to its marketing approval. Chemistry manufacturing and controls, animal toxicology, and biopharmaceutical data are described. Results of Phase I and Phase II clinical studies in patients with CML in blast crisis (CML-BC), in accelerated phase (CML-AP), and in chronic phase disease-resistant or intolerant to IFN-alpha (CML-CP) are summarized. The basis for marketing approval and postmarketing commitments by the pharmaceutical company are discussed. Toxicology studies in the rat, dog, and monkey show the hematological, renal, and hepatobiliary toxicity of imatinib. Pharmacokinetic studies in patients with CML demonstrate 98% imatinib bioavailability. The elimination half-lives of the parent drug and the major active metabolite, CGP74588, from plasma are approximately 18 and 40 h, respectively. Approximately 81% of the drug is eliminated in 7 days, 68% in the feces and 13% in the urine. Cytochrome P-450 3A4 is the main enzyme responsible for imatinib metabolism. Phase I and II clinical studies were conducted. The Phase I study, in 83 CML patients, evaluated oral imatinib doses from 25 to 1000 mg/day. Dose-limiting toxicity was not observed. The three Phase II studies, in CML-CP, CML-AP, and CML-BC, enrolled 1027 patients. CML-CP patients received 400 mg/day imatinib, whereas CML-AP and CML-BC patients generally received 600 mg/day imatinib. Primary study endpoints were cytogenetic response rate (CML-CP) and hematological response rate (CML-AP and CML-BC). The cytogenetic response rate for CML-CP patients was 49%. The hematological response rate of CML-AP and CML-BC patients was 63 and 26%, respectively. The most common imatinib adverse events were nausea, vomiting, myalgia, edema, and diarrhea. Elevated liver enzymes and/or bilirubin were reported in 27 patients (2.6%). On May 10, 2001, imatinib mesylate (Gleevec, formerly known as STI-571 and Glivec), manufactured and distributed by Novartis Pharmaceuticals, East Hanover, NJ, was approved by the United States Food and Drug Administration for the treatment of CML in three clinical settings: CML-BC, CML-AP, and CML-CP. This report summarizes the Food and Drug Administration's review of the New Drug Application.


Handheld Tissue Oximetry for the Prehospital Detection of Shock and Need for Lifesaving Interventions: Technology in Search of an Indication?
Improved prehospital methods for assessing the need for lifesaving interventions (LSIs) are needed to gain critical lead time in the care of the injured. We hypothesized that threshold values using prehospital handheld tissue oximetry would detect occult shock and predict LSI requirements. This was a prospective observational study of adult trauma patients emergently transported by helicopter. Patients were monitored with a handheld tissue oximeter (InSpectra Spot Check; Hutchinson Technology Inc, Hutchinson, MN), continuous vital signs, and 21 laboratory measurements obtained both in the field with a portable analyzer and at the time of admission. Shock was defined as base excess ≥ 4 or lactate > 3 mmol/L. Eighty-eight patients were enrolled with a median Injury Severity Score of 16 (interquartile range, 5-29). The median hemoglobin saturation in the capillaries, venules, and arterioles (StO2) value for all patients was 82% (interquartile range, 76%-87%; range, 42%-98%). StO2 was abnormal (< 75%) in 18 patients (20%). Eight were hypotensive (9%) and had laboratory-confirmed evidence of occult shock. StO2 correlated poorly with shock threshold laboratory values (r = -0.17; 95% confidence interval, -0.33 to 1.0; P = .94). The area under the receiver operating curve was 0.51 (95% confidence interval, 0.39-0.63) for StO2 < 75% and laboratory-confirmed shock. StO2 was not associated with LSI need on admission when adjusted for multiple covariates, nor was it independently associated with death. Handheld tissue oximetry was not sensitive or specific for identifying patients with prehospital occult shock. These results do not support prehospital StO2 monitoring despite its inclusion in several published guidelines.


Evaluation of systemic inflammatory responses in neonates with herpes simplex virus infection.
Neonatal herpes simplex virus (HSV) infection is a severe disease with high mortality and morbidity. To investigate the pathogenesis of neonatal HSV infection, we examined inflammatory responses and markers of apoptosis in patients with neonatal HSV infection. Concentrations of inflammatory cytokines and markers of apoptosis were significantly higher in patients with disseminated HSV infection and were correlated with HSV load. It appears that the immunopathological damage that results from host responses to viral infection leads to organ dysfunction in patients with neonatal HSV infection.


Characteristics, management and outcome of patients with recurrent tuberculosis under routine programme conditions in Malawi.
All 43 non-private hospitals in Malawi, which registered TB cases between 1 July 1999 and 30 June 2000. To determine 1) the characteristics, management and treatment outcome, 2) timing of the previous episode of TB, and 3) pattern of drug resistance in patients registered with recurrent smear-positive pulmonary TB. Retrospective data collection using TB registers and laboratory culture and drug sensitivity registers. There were 748 recurrent patients; data were available for 747. Of these, 487 (65%) successfully completed a re-treatment regimen, 185 (25%) died and the remainder had another outcome. Information about previous TB was recorded for 491 (66%) patients. In 286 (58%) there were 2 years or less between completing and re-starting treatment. Only 307 (41%) patients had sputum sent for culture and drug sensitivity tests. In 164 patients with cultures of Mycobacterium tuberculosis, 122 (81%) were fully sensitive, 25 (15%) had resistance to isoniazid and/or streptomycin, and 6 (4%) had resistance to isoniazid and rifampicin (MDR-TB). Patients with recurrent TB had acceptable treatment outcomes, and most had fully sensitive organisms. Over half had recurrent TB 2 years or less after completing treatment. Ways to prevent recurrence need to be investigated and implemented in the field.


Screening for Cystic Fibrosis Related Complications in the Context of a Pandemic and Altered Models of Care.
Standard of care recommend that patients with cystic fibrosis (CF) require screening investigations to assess for complications. Changing models of care due to the COVID19 pandemic may have impacted completion of recommended screening. To compare the frequency of screening investigations completed in people with CF before and after the onset of the COVID19 pandemic. Medical records were reviewed at 4 CF-specialist centers to identify screening investigations completed in the 12-months before and after pandemic onset. Records of 625 patients were reviewed. Prior to pandemic onset, there was between center variability in completion of screening investigations. There was greatest baseline variation between centers in performing oral glucose tolerance test (OGTT); range 38%-69%, exercise tests; 3%-51% and sputum screening for non-tuberculous mycobacteria; 53%-81%. Following pandemic onset, blood tests, and sputum cultures were maintained at the highest rates. Exercise testing, CXR and OGTT exhibited the greatest declines, with reductions at individual centers ranging between 10%-24%, 22%-43%, and 20%-26%, respectively. Return to in-person visits following pandemic onset was variable, ranging from 16% to 74% between centers. Completion of screening investigations varies between CF centers and changes in models of care, such as increased virtual care in response to COVID19 pandemic was associated with reduction in completion of investigations. Centers would benefit from auditing their adherence to standards of care, particularly considering recent changes in care delivery.


Debate: endoscopy is unnecessary in the management of uncomplicated GERD.
The case of a 56-year-old man with recurrent retrosternal heartburn no longer relieved by antacids is discussed. Arguments for and against conducting endoscopy in this patient are presented. Initial therapy with a standard dose proton pump inhibitor, without endoscopy is the suggested treatment strategy. The main purpose of conducting an endoscopy in a patient with chronic gastrointestinal reflux is to detect the presence of Barrett's esophagus. However, data indicate that the presence of Barrett's esophagus is unrelated to symptoms and that it is not significantly associated with heartburn. Additionally, there is no certainty that survival is improved by the detection of Barrett's esophagus. Nonetheless, many clinical practice guidelines support conducting endoscopy in patients with GERD.


Magnetic resonance imaging with gadolinium-DTPA for detecting cardiac transplant rejection in rats.
To date, no noninvasive tool has gained widespread acceptance as an adequate substitute for endomyocardial biopsy for the diagnosis and grading of cardiac transplant rejection. We examined the potential role of magnetic resonance imaging with gadolinium (Gd)-diethylenetriamine penta-acetic acid (DTPA) image enhancement for the diagnosis of cardiac graft rejection. We studied 15 rats with heterotopic cardiac transplants, nine of which received no immunosuppression, and six of which received cyclosporine, azathioprine, and methylprednisolone. The animals underwent magnetic resonance imaging, which was immediately followed by sacrifice (2-12 days after transplant). Myocardial image enhancement was assessed on T1-weighted images performed before and after administration of Gd-DTPA, 0.5 mmol/kg. Histological specimens were graded I, II, or III to indicate increasing severity of rejection. In the absence of rejection, Gd-DTPA induced mild homogeneous myocardial enhancement. Ten of 11 cases with Grade II or III rejection manifested one or more areas of intense myocardial enhancement. The extent and distribution of intense myocardial enhancement corresponded to the severity and distribution of histological rejection. Quantitative myocardial enhancement, expressed as the ratio of maximal signal intensity after Gd-DTPA to signal intensity before Gd-DTPA administration, separated Grade I animals (1.61 +/- 0.27; mean +/- SD) from Grades II (2.89 +/- 0.58) and III (3.10 +/- 0.77; p less than 0.01) animals. In conclusion, cardiac transplant rejection is characterized by intense T1-weighted image enhancement after administration of Gd-DTPA. Magnetic resonance imaging with Gd-DTPA thus has potential application in the clinical diagnosis of cardiac transplant rejection.


Gender Disparities in Hypertensive Emergency Admissions: A National Retrospective Cohort Study.
Background Hypertension is one of the most common conditions affecting almost one in every five adults globally and hypertensive emergency is a life-threatening complication of uncontrolled hypertension leading to significant disability. Despite advances in treatment, gender disparities are yet to be addressed. Methods This retrospective cohort study used nationally representative data from the Healthcare Cost and Utilization Project (HCUP), specifically the National Inpatient Sample, to study two cohorts divided by sex (males and females). The primary outcome was all-cause inpatient mortality. Multivariate logistic regression analysis yielded adjusted odds ratios (aORs) for confounders. Secondary outcomes included length of stay (LOS) and total hospital charges. Multivariate linear regression identified independent predictors. We described crude rates of mechanical ventilation, acute kidney injury (AKI) requiring hemodialysis (HD), and vasopressor requirements. Patient demographics were also presented. We used the chi-squared (χ2) test for categorical variables and Student's t-test for continuous variables. Statistical significance was defined as a two-tailed p-value<0.05. Results A total of 229,025 patients met the inclusion criteria, where 52% were male and 48% were female. The mean patient age was 58 years (55 for men and 62 for women, p <0.001). White patients represented 40% of hospitalizations (males: 37%; females: 42%), black patients represented 42% (males: 43%; females: 41%), and Hispanics 11% (males: 12%; females: 10%). Medicare was the primary payer 47% of the time (males: 38%; females: 56%), Medicaid in 21% (males: 23%; females: 18%), private insurance in 20% (males: 23%; females: 17%), and no insurance in 10% (males: 14%; females: 7%). Female patients had higher rates of chronic obstructive pulmonary disease (COPD) (21% for females vs. 15% for males), connective tissue disease (4.6% for females vs. 0.98% for males; p<0.001), and dementia (6% for females vs. 3% for males). Conversely, males had a higher rate of chronic kidney disease (CKD) (51% vs. 42% for females). Male sex was a predictor of mortality (aOR 1.39, p=0.036), along with age (aOR 1.02, p<0.001) and Charlson Comorbidity Index (http://mchp-appserv.cpe.umanitoba.ca/viewConcept.php?printer=Y&conceptID=1098) (aOR 1.20, p<0.001). Sex was not a predictor of length of stay (LOS) (p=0.496) or total hospital charges (p=0.192). Conclusions In an attempt to achieve better outcomes in patients affected by hypertensive emergency, our retrospective cohort study found that male patients who experienced hypertensive emergency had 39% higher odds of mortality than female patients. Age and Charlson Comorbidity Index were additionally found to be independent predictors of mortality.


Variation in the behavioral responses of Budgerigars Melopsittacus undulatus to an alarm call in relation to sex and season.
In the Budgerigar Melopsittacus undulatus, the significance of a conspecific alarm call was investigated in two seasons, winter and spring. Two qualitatively different behavioral responses were displayed by the receivers in reply to playbacks: call(s) and/or taking flight(s). The comparative analysis of the number of birds responding to the alarm and to the control signals revealed two major facts: 1) in both seasons, the responses to the alarm signal were only observed for females, not for males, 2) qualitatively, females exhibited a great inter-season variability in their behavioral responses to the hearing of an alarm call. In winter, the females were more predisposed to emit acoustic responses while in spring they mainly took flight.


Biomarkers reflect differences in osteoarthritis phenotypes of the lumbar spine: the Johnston County Osteoarthritis Project.
To determine differences in biomarker levels between radiographic phenotypes of facet joint osteoarthritis (FOA) only, spine OA only ((disc space narrowing (DSN) and vertebral osteophytes (OST)) or the combination of FOA and spine OA. A cross-sectional analysis of data from 555 participants in the Johnston County Osteoarthritis Project was performed. Lumbar spine levels were graded by severity (OST and DSN) and presence (FOA) of degeneration. Biomarkers included hyaluronan (HA) and type II collagen (CTX-II). Adjusted risk ratios (aRRR) were estimated using multinomial regression, with adjustment for age, race, sex, body mass index (BMI), and radiographic OA (knee, hip, hand). Interactions were tested between sex, race and low back symptoms. FOA only was present in 22.4%, 14.5% had spine OA only, and 34.6% had the combination of FOA and spine OA. Compared to the reference group of neither FOA or spine OA, a one unit higher ln HA level was associated with 31% higher relative risk ratio (RRR = 1.31 (95% 1.03, 1.67)) of having FOA only, while, a one unit higher lnuCTX-II level was associated with 84% higher relative risk ratio (RRR = 1.84 (95% CI 1.19, 2.84)) of having spine OA only. No significant interactions were identified. Interestingly, OA affecting the synovial facet joint was associated with a marker of inflammation (HA). Spine OA, affecting intervertebral discs that contain collagen type II, was associated with a marker reflecting collagen type II degradation (CTX-II). These findings suggest that biomarkers may reflect the different pathophysiologic processes of lumbar spine OA phenotypes.


Labeling and intergenerational transmission of crime: The interaction between criminal justice intervention and a convicted parent.
Labeling theory suggests that criminal justice interventions amplify offending behavior. Theories of intergenerational transmission suggest why children of convicted parents have a higher risk of offending. This paper combines these two perspectives and investigates whether labeling effects might be stronger for children of convicted parents. We first investigated labeling effects within the individual: we examined the impact of a conviction between ages 19-26 on self-reported offending behavior between 27-32 while controlling for self-reported behavior between 15-18. Our results show that a conviction predicted someone's later self-reported offending behavior, even when previous offending behavior was taken into account. Second, we investigated whether having a convicted parent influenced this association. When we added this interaction to the analysis, a labeling effect was only visible among people with convicted parents. This supports the idea of cumulative disadvantage: Labeling seems stronger for people who are already in a disadvantaged situation having a convicted parent.


Mechanistic issues in asparagine synthetase catalysis.
The enzymatic synthesis of asparagine is an ATP-dependent process that utilizes the nitrogen atom derived from either glutamine or ammonia. Despite a long history of kinetic and mechanistic investigation, there is no universally accepted catalytic mechanism for this seemingly straightforward carboxyl group activating enzyme, especially as regards those steps immediately preceding amide bond formation. This chapter considers four issues dealing with the mechanism: (a) the structural organization of the active site(s) partaking in glutamine utilization and aspartate activation; (b) the relationship of asparagine synthetase to other amidotransferases; (c) the way in which ATP is used to activate the beta-carboxyl group; and (d) the detailed mechanism by which nitrogen is transferred.


[Evaluation of the QBC-Autoread System in hematological screening in a hospital laboratory].
The QBC Autoread System is based on fully automatic, electro-optical linear measurements of the discrete layers of packed blood cells in a microhematocrite tube. The layering results from differing cell densities during high-speed centrifugation. The determination of hemoglobin, hematocrit, leucocytes, platelets and of the percentual share of granulocytes and lymphocytes/monocytes with the QBC Autoread System was evaluated concerning reliability and precision in comparison with the conventional analytical methods. We used a Coulter Counter System and the manual differentiation of the stained blood smears as reference methods. The evaluation of the first 400 blood samples showed a close correlation for all hematological parameters examined, but no significant deviation relative to the reference methods. Only 83 of totally 1000 (8.3%) examined venous blood samples could not be evaluated completely by the QBC Autoread System. For the analysis of very pathological cell populations, especially of hematological oncological diseases, the QBC System is not qualified. The extreme portability, the quick practicability and the good accordance with the conventional analytical methods make the QBC Autoread System a qualified screening method for practice and emergency laboratories.


[Human embryonic haemoglobins. Ac-Ser-Leu-Thr-is the N-terminal sequence of the zeta-chains (author's transl)].
The complete amino acid sequence of the embryonic zeta-chains has been reported in an earlier communication (Aschauer, H., Sanguansermsri, T. & Braunitzer, G. (1981), Hoppe-Seyler's Z. Physiol. Chem. 362, 1159-1162). To elucidate the nature of the N-terminal blocking group, tryptical cleavage of the zeta-chains was carried out and the N-terminal tryptic tetrapeptide was isolated by high performance liquid chromatography on reversed phase (RP 8). The tetrapeptide was digested with pronase and the mixture was subjected to mass spectrometry by chemical ionization. By this method Ac-Ser was found to be the N-terminus. In a second experiment the tetrapeptide was permethylated and analysed by mass spectrometry with electron ionization. The N-terminal sequence was found to be Ac-Ser-Leu-Thr-.


Association of insulinoma with subcutaneous nodular fat necrosis.
A 57-year-old man first noted tender subcutaneous nodules on his extremities and abdomen. Six months later symptoms of insulin hypersecretion became evident and hyperinsulinemia was documented. An exploratory laparotomy revealed an islet cell tumor measuring 12 cm in diameter extending anteriorly from head of the pancreas with metastases in the liver. Evidence of parathyroid and pituitary hormone hypersecretion suggests that the insulinoma may be part of a multiple endocrine neoplasia syndrome. Histological examination of the subcutaneous nodules revealed the characteristic histology of nodular fat necrosis. This is the first report of the association of an insulin-secreting islet cell carcinoma with subcutaneous nodular fat necrosis.


Critical 23S rRNA interactions for macrolide-dependent ribosome stalling on the ErmCL nascent peptide chain.
The nascent peptide exit tunnel has recently been identified as a functional region of ribosomes contributing to translation regulation and co-translational protein folding. Inducible expression of the erm resistance genes depends on ribosome stalling at specific codons of an upstream open reading frame in the presence of an exit tunnel-bound macrolide antibiotic. The molecular basis for this translation arrest is still not fully understood. Here, we used a nucleotide analog interference approach to unravel important functional groups on 23S rRNA residues in the ribosomal exit tunnel for ribosome stalling on the ErmC leader peptide. By replacing single nucleobase functional groups or even single atoms we were able to demonstrate the importance of A2062, A2503 and U2586 for drug-dependent ribosome stalling. Our data show that the universally conserved A2062 and A2503 are capable of forming a non-Watson-Crick base pair that is critical for sensing and transmitting the stalling signal from the exit tunnel back to the peptidyl transferase center of the ribosome. The nucleobases of A2062, A2503 as well as U2586 do not contribute significantly to the overall mechanism of protein biosynthesis, yet their elaborate role for co-translational monitoring of nascent peptide chains inside the exit tunnel can explain their evolutionary conservation.


Sexual self-esteem in mothers of normal and mentally-retarded children.
Sexual self-esteem is negatively influenced by the stressful experiences in lifetime. This study compared the sexual self-esteem and its components in mothers with normal and mentally-retarded children in Qaen city, in 2014. A total of 120 mothers were selected and assigned into two groups of 60 samples based on convenient sampling method and randomized multiple stage sampling. Both groups completed sexual self-esteem questionnaire. The data were analyzed employing t-test through SPSS software version15. The results showed that the rate of sexual self-esteem in mothers of mentally-retarded children decreased significantly compared with that of mothers with normal children (p<0.05). Moreover, the mean scores of all components of sexual self-esteem including skill and experience, attractiveness, control, moral judgment, and adaptiveness in mothers of mentally-retarded children were significantly less than those of mothers with normal children (p <0.05). Therefore, it is recommended that self-esteem, especially the sexual one, be taught to mothers of mentally-retarded children by specialists.


[Induced abortion in France: new legislation and questions from healthcare providers].
The latest changes in French law concerning induced abortion as well as comments from healthcare providers are discussed here. The new articles of the French law are explained. Some of the changes are to be applied immediately: limit set at 14 weeks amenorrhea, non-mandatory psychological consultation, referent adult. Other modifications are pending, including ambulatory induced abortion. These different changes raise many questions.


Autoimmunity in myocarditis: relevance of animal models.
Infections (viruses, bacteria, protozoa, fungi) are major etiological factors causing clinical myocarditis and dilated cardiomyopathy. In many patients and symptom-free relatives antibodies and T cells reactive to heart antigens are detected, which implies that autoimmunity is probably a major pathogenic mechanism of cardiac injury. Animal models have been established to elucidate how infections initiate autoimmunity and how autoimmune mediators cause death or transient dysfunction of myocytes. Two major types of experimental models are discussed: adjuvant-induced myocarditis, in which animals are given multiple immunizations of heart proteins (myosin, adenine nucleotide translocator); and virus-induced myocarditis, in which animals are infected with the viruses predominantly associated with the human disease.


Telomere length and somatic mutations in correlation with response to immunosuppressive treatment in aplastic anaemia.
We investigated the frequencies of cytogenetic aberrations and somatic mutations of prognostic relevance in 393 patients with aplastic anaemia (AA). Clonality was determined by G-banding/fluorescence in situ hybridization (FISH) (n = 245), and targeted capture sequencing was performed for 88 haematopoiesis-related genes (n = 70). The telomere length (TL) of bone marrow nucleated cells was measured at the single cell level by FISH (n = 135). Eighteen (4·6%) patients showed disease progression, and monosomy 7 (50·0%) was the most predominant cytogenetic evolution at disease transformation. One third of patients (32·9%) presented at least 1 mutation; the most frequently mutated genes were NOTCH1, NF1, SCRIB, BCOR and DNMT3A. The patient group with clonal changes (30·7%) showed an adverse response to immunosuppressive treatment (IST), compared to the non-clonal group, but this finding did not show statistical significance. The TL of AA patients was significantly shorter than normal control and patients with clonal changes showed significantly shorter TLs. Patients with TL>5·9 showed a higher response rate to IST (P = 0·048). In conclusion, the patients with clonal changes or TL attrition showed a poor response to IST. Shorter TL can be used not only as a biomarker, but also as a predictive marker for treatment response to IST.


The influence of suction catheter tip design on tracheobronchial trauma and fluid aspiration efficiency.
The suctioning efficiency and trauma-producing characteristics of five commercially available tracheobronchial suction catheters (Pharmaseal Tri-Flo, NCC Gentle-Flo, Argyle Aero-Flo, Argyle Dual Side-Hole, and Pharmaseal Whistle-Tip) were experimentally evaluated in anesthetized healthy dogs. The tendency of catheters to invaginate or "grab" tracheobronchial mucosa was observed with a bronchofiberscope during suctioning. Mucosal grabbing was seldom seen even at high (greater than 300 torr) vacuum levels with the cateter tip in the trachea. All catheters were observed to invaginate mucosa in lobar and segmental bronchi, with the frequency of grabbing being a function of airway anatomy, airway size, catheter orientation, tip design, and vacuum level. Catheters with multiple side-holes appeared to invaginate mucosa less frequently than the single side-hole catheter. Repeated suctioning of anesthetized healthy dogs followed by tracheobronchial excision, gross observation, and histologic examination of mucosal tissue biopsies demonstrated significant differences in the frequency and severity of lesions caused by the tracheobronchial suction procedure. All catheters were observed to damage airway lining, the damage related to multiple side-hole catheters appearing to be associated entirely with the act of cateter insertion and not with the application of vacuum. Only the Whistle-Tip design produced measurable damage beyond that related to catheter insertion. The average tip-suctioning effectiveness for each catheter, determined in vitro by aspirating a thin, uniform layer of simulated mucus, was found to be significantly higher for the Tri-Flo and Whistle-Tip catheters than the others, the Aero-Flo being least effective. Preliminary attempts to demonstrate this difference in suctioning effectiveness by comparing the performance of the catheters which displayed the highest and lowest tip suction effectiveness in a standardized clinical suctioning procedure revealed no significant difference in the percentage of mucus removed by either catheter. Additional studies should clarify this apparent contradiction.


Racial discrimination and metabolic control in women with type 2 diabetes.
We investigated whether self-reported racial discrimination was associated with insulin resistance (IR) and glycosylated hemoglobin (A1c) in women with type 2 diabetes in the United States, after controlling for covariates. Seventy-seven Black and White women with type 2 diabetes completed the Experiences of Discrimination Scale, which assesses self-reported lifetime frequency of racially motivated discrimination. Participants provided fasting blood samples for assessment of glucose and insulin for determination of IR and A1c. Covariates included age, education, waist circumference, diabetes distress, and stressful life events. In unadjusted regression analysis discrimination was significantly associated with IR. There was a trend for a race by discrimination interaction, with a weaker effect for Blacks than Whites. Follow up analysis showed that discrimination was significantly associated with IR in both Blacks and Whites, even after adjustment, as was waist circumference. In unadjusted regression analysis, discrimination was significantly associated with A1c. There was a significant race by discrimination interaction. Follow up analysis showed that discrimination was not significantly associated with A1c among Blacks, but was among Whites, even after adjustment, as was diabetes distress and insulin use. Racial discrimination is associated with insulin resistance in Black and White women with diabetes, and with A1c in White women with diabetes.


TPI1-reduced extracellular vesicles mediated by Rab20 downregulation promotes aerobic glycolysis to drive hepatocarcinogenesis.
Rab GTPases are major mediators that ensure the proper spatiotemporal regulation of intracellular trafficking. Functional impairment and altered expression of Rab proteins have been revealed in various human cancers. There is an emerging evidence about the role of Rab proteins in the biogenesis of extracellular vesicles (EVs). In hepatocellular carcinoma (HCC), using RNA sequencing comparing expression profiles of adjacent non-tumorous tissues and HCC, Rab20 is identified to be the most frequently downregulated Rab member in HCC. Functionally, restoration of Rab20 in metastatic HCC cells results in the release of EVs with a diminished activity to promote cell growth, motility and metastasis. Conversely, EVs released from normal liver cells with Rab20 knockdown loses suppressive effect on HCC cell growth and motility. Proteomic profiling revealed the level of triosephosphate isomerase 1 (TPI1), a glycolytic enzyme, in EVs to be positively associated with Rab20 expression of the releasing cells. TPI1 targeted to be expressed in EVs released by Rab20 knockdown cells compromises the oncogenic activity of EVs. Besides, EVs released by TPI1 knockdown cells recapitulates the promoting effect of EVs derived from HCC cells with Rab20 underexpression. Aerobic glycolysis is beneficial to the survival and proliferation of tumour cells. Here, we observed that the enhanced cell growth and motility are driven by the enhanced aerobic glycolysis induced by EVs with reduced TPI1. The addition of glycolytic inhibitor blocks the promoting effect of EVs with reduced TPI1. Taken together, our study provides a mechanistic link among tumour cell-derived EVs and glucose metabolism in HCC with Rab20 deregulation.


Axitinib for the treatment of advanced non-small-cell lung cancer.
Lung cancer is the most frequently diagnosed cancer in men and comprises 23% of total cancer deaths worldwide. The majority of patients present with advanced disease, for whom the 5-year survival is < 5%. Since angiogenesis plays a central role in tumourigenesis, inhibiting this pathway may improve outcomes in non-small-cell lung cancer (NSCLC). Axitinib is one of the latest and most potent anti-angiogenic tyrosine kinase inhibitors (TKI) currently being evaluated to treat NSCLC. In this review, the rationale for targeting angiogenesis in lung cancer, other angiogenic agents in NSCLC, axitinib's mechanism of action, pharmacology and metabolism, and the preclinical and clinical data to date in NSCLC will be discussed. Several TKI which target angiogenesis pathways have resulted in improved response rates and progression-free survival in NSCLC, but no improvement in overall survival in clinical trials. Axitinib is a more potent inhibitor of vascular endothelial growth factor receptors than other TKI, but this has yet to translate into a clinical benefit. Phase II trials are ongoing, but the published data to date has yet to support a role for axitinib in the treatment algorithm for NSCLC.


Faecal indicator bacteria and antibiotic-resistant β-lactamase producing Escherichia coli in blackwater: a pilot study.
The aim of this study was to identify and quantify faecal indicator bacteria in blackwater collected from a source separation unit and determine the amount of E. coli isolates resistant to antimicrobials and their potential to produce extended spectrum β-lactamases (ESβLs) and metallo-β-lactamases (MβLs), which hydrolyse the most important antibiotics used in clinical practice. Most of the isolates were resistant to amoxicillin with clavulanic acid (36.4 %), followed by ticarcillin with clavulanic acid (22.7 %) and tetracycline (18.2 %). ESβL-producing genes blaCTX-M and blaTEM were found in three (13.6 %) and four (18.2 %) E. coli strains, respectively, while MβL genes were found in two (9.1 %). By separating at source, this pilot study clearly shows that gastrointestinal bacteria of healthy people can be an important source of antibiotic resistance released into the environment through wastewaters. One way to prevent that is to treat wastewater with a combination of TiO2, UV light, or ozone, as successful methods to remove resistant bacteria and prevent their spread in the environment.


Three-dimensional high-definition flow in the diagnosis of placental lakes.
Placental lakes are sonolucent areas often found in the normal placenta. Most of them are asymptomatic. They are sometimes related to placenta accreta or intrauterine fetal growth restriction, among other conditions. Although Doppler sonography is useful for evaluating noxious placental lakes, it is not easy to adapt Doppler studies to conventional two-dimensional color Doppler sonography because of the low-velocity blood flow and high vascularity in the placenta. Here, we demonstrate how three-dimensional high-definition imaging of flow provides a novel visual depiction of placental lakes, which helps substantially with the differential diagnosis. As far as we know, there have been no previous reports of observation of placental lakes using three-dimensional high-definition imaging of flow.


Human dermal fibroblasts exhibit delayed adipogenic differentiation compared with mesenchymal stem cells.
Human dermal fibroblasts (FBs) express mesenchymal stem cell (MSC)-specific cell surface markers and differentiate into several cell types under appropriate conditions. Molecular mechanisms controlling the early stages of differentiation of dermal FBs and MSCs isolated from different sources have not been well studied. Here, we have analyzed the cell type-specific changes of adipose tissue-derived mesenchymal stem cells (AdMSCs) and dermal FBs in the process of differentiation into adipocytes and osteoblasts. Analysis of gene expression in the course of adipogenic differentiation of AdMSCs and FBs isolated from the same individuals revealed a time lag in the induction of adipogenesis-related genes in FBs compared with AdMSCs, a phenomenon not previously described. Further, preliminary evidence suggests that delayed adipogenesis of FBs is related to the delayed induction of preadipocyte transcription factor ZNF423 in FBs. These findings clearly show that AdMSCs and FBs have similar developmental potential but different molecular control mechanisms of initial stages of adipogenic differentiation.


Accelerated 2D Cartesian MRI with an 8-channel local B0 coil array combined with parallel imaging.
In MRI, the magnetization of nuclear spins is spatially encoded with linear gradients and radiofrequency receivers sensitivity profiles to produce images, which inherently leads to a long scan time. Cartesian MRI, as widely adopted for clinical scans, can be accelerated with parallel imaging and rapid magnetic field modulation during signal readout. Here, by using an 8-channel local B 0 $$ {\mathrm{B}}_0 $$ coil array, the modulation scheme optimized for sampling efficiency is investigated to speed up 2D Cartesian scans. An 8-channel local B 0 $$ {\mathrm{B}}_0 $$ coil array is made to carry sinusoidal currents during signal readout to accelerate 2D Cartesian scans. An MRI sampling theory based on reproducing kernel Hilbert space is exploited to visualize the efficiency of nonlinear encoding in arbitrary sampling duration. A field calibration method using current monitors for local B 0 $$ {\mathrm{B}}_0 $$ coils and the ESPIRiT algorithm is proposed to facilitate image reconstruction. Image acceleration with various modulation field shapes, aliasing control, and distinct modulation frequencies are scrutinized to find an optimized modulation scheme. A safety evaluation is conducted. In vivo 2D Cartesian scans are accelerated by the local B 0 $$ {\mathrm{B}}_0 $$ coils. For 2D Cartesian MRI, the optimal modulation field by this local B 0 $$ {\mathrm{B}}_0 $$ array converges to a nearly linear gradient field. With the field calibration technique, it accelerates the in vivo scans (i.e., proved safe) by threefold and eightfold free of visible artifacts, without and with SENSE, respectively. The nonlinear encoding analysis tool, the field calibration method, the safety evaluation procedures, and the in vivo reconstructed scans make significant steps to push MRI speed further with the local B 0 $$ {\mathrm{B}}_0 $$ coil array.


Characterization and expression of an Fc gamma receptor cDNA cloned from rat natural killer cells.
A cDNA clone for an IgG-binding Fc receptor, rtFc gamma R alpha, of the rat natural killer cell line CRNK-16 is characterized here. This clone encodes an Fc gamma receptor as shown by the ability of cDNA-transfected COS cells to rosette IgG-coated sheep erythrocytes. The rtFc gamma R alpha is exceptionally homologous to the mouse moFc gamma R alpha, with 77% protein sequence identity and 71% nucleic acid identity overall. The transmembrane region of the rtFc gamma R alpha contains the sequence Leu-Phe-Ala-Val-Asp-Thr-Gly-Leu, which is present in the membrane sequences of four other Fc receptors including mouse Fc gamma R alpha, human Fc gamma RIII-2, and the Fc epsilon R alpha subunits of the rat and human high-affinity IgE-binding receptors. Also, the rtFc gamma R alpha cytoplasmic domain exhibits specific homology to other receptors derived from natural killer cells, human Fc gamma RIII-2 and mouse Fc gamma R alpha. However, the rtFc gamma R alpha cDNA clone is complementary to at least two different-sized mRNAs expressed by CRNK-16 cells, contrasting the single Fc gamma R-related mRNA species expressed by human and mouse natural killer cells. These rat mRNAs are homologous to both the 5' and the 3' end of the cDNA clone, suggesting that they may be (i) splice variants of one transcript or (ii) products of different but highly related genes.


Vision following extended congenital blindness.
Animal studies suggest that early visual deprivation can cause permanent functional blindness. However, few human data on this issue exist. Given enough time for recovery, can a person gain visual skills after several years of congenital blindness? In India, we recently had an unusual opportunity to work with an individual whose case history sheds light on this question. S.R.D. was born blind, and remained so until age 12. She then underwent surgery for the removal of dense congenital cataracts. We evaluated her performance on an extensive battery of visual tasks 20 years after surgery. We found that although S.R.D.'s acuity is compromised, she is proficient on mid- and high-level visual tasks. These results suggest that the human brain retains an impressive capacity for visual learning well into late childhood. They have implications for current conceptions of cortical plasticity and provide an argument for treating congenital blindness even in older children.


Bcl-2 overexpression does not enhance in vivo axonal regeneration of retinal ganglion cells after peripheral nerve transplantation in adult mice.
Optic nerve (ON) injury in adult mammals causes retinal ganglion cell (RGC) death and subsequent visual loss. Recovery of vision requires both rescuing axotomized RGCs and inducing their axonal regeneration. Axotomized RGCs are significantly rescued by overexpression of bcl-2, an anti-apoptotic gene. However, whether bcl-2 affects axonal regeneration is controversial. In neonatal bcl-2 transgenic mice (bcl-2 mice), optic tract regeneration after tectal lesion was promoted (Chen et al., 1997), whereas ON regeneration after ON crush was not (Lodovichi et al., 2001). These conflicting results may be attributable to different environments between tectum and ON. We tested here whether bcl-2 overexpression enhances in vivo RGC axonal regeneration in adult mice through a permissive environment in the peripheral nerve (PN) graft. Four weeks after PN transplantation to the proximal ON stump, we assessed the number of surviving and regenerating RGCs by retrograde labeling. Although the survival rate in bcl-2 mice was significantly enhanced compared with that in wild-type (wt) mice, the regeneration rate was not enhanced. In both bcl-2 and wt mice, RT97 immunostaining of the PN-grafted retinas revealed some RGC axons regrowing intraretinally but repulsed at the optic disk. To circumvent this repulsive barrier, we directly transplanted the PN graft to the partially injured retina and compared regeneration rates between these mice. Here again the regeneration rate in bcl-2 mice did not exceed that in wt mice. These findings indicate that bcl-2 overexpression enhances survival but not axonal regeneration of adult RGCs even within a permissive environment.


Structure and properties of alpha-synuclein and other amyloids determined at the amino acid level.
The structure of alpha-synuclein (alpha-syn) amyloid was studied by hydrogen-deuterium exchange by using a fragment separation-MS analysis. The conditions used made it possible to distinguish the exchange of unprotected and protected amide hydrogens and to define the order/disorder boundaries at close to amino acid resolution. The soluble alpha-syn monomer exchanges its amide hydrogens with water hydrogens at random coil rates, consistent with its natively unstructured condition. In assembled amyloid, long N-terminal and C-terminal segments remain unprotected (residues 1- approximately 38 and 102-140), although the N-terminal segment shows some heterogeneity. A continuous middle segment (residues approximately 39-101) is strongly protected by systematically H-bonded cross-beta structure. This segment is much too long to fit the amyloid ribbon width, but non-H-bonded amides expected for direction-changing loops are not apparent. These results and other known constraints specify that alpha-syn amyloid adopts a chain fold like that suggested before for amyloid-beta [Petkova et al. (2002) Proc. Natl. Acad Sci. USA 99, 16742-16747] but with a short, H-bonded interlamina turn. More generally, we suggest that the prevalence of accidental amyloid formation derives mainly from the exceptional ability of the main chain in a structurally relaxed beta-conformation to adapt to and energy-minimize side-chain mismatching. Seeding specificity, strain variability, and species barriers then arise because newly added parallel in-register chains must faithfully reproduce the same set of adaptations.


Molecular mechanisms responsible for the involvement of tissue transglutaminase in human diseases: Celiac Disease.
Tissue transglutaminase (tTG or TG2; E.C. 184.108.40.206) belongs to the transglutaminase family, a group of closely related enzymes that share the ability to catalyze the cross-linking of a glutaminyl residue of a protein/peptide substrate to a lysyl residue of a protein/peptide co-substrate. tTG is a multifunctional enzyme since it is also capable of catalyzing other biochemical reactions. The distribution and physiological roles of tTG have been widely studied in numerous cell types and tissues, but only recently its role in human diseases has started to be clarified. For example, transglutaminase activity has been hypothesized to be involved in the pathogenetic mechanisms responsible for several human diseases, including neurodegenerative diseases, such as polyglutamine diseases hitherto identified. Among human diseases, a large and recent series of studies have clearly shown that the activity of the tTG is critical for a very diffuse human pathology known as Celiac Disease. This disease is due to intolerance to a food component, gliadin, and is characterized by a very complex clinical syndrome, including gastrointestinal pathological manifestations, often associated with extra-intestinal manifestations. Interestingly, a subset of celiac patients also develops certain neurological disorders. In this review we describe the roles played by tTG in the molecular mechanisms responsible for pathophysiology of Celiac Disease.


Delivery of CW laser power up to 300 watts at 1080 nm by an uncooled low-loss anti-resonant hollow-core fiber.
In this paper, we report the use of a 3-meter low-loss anti-resonant hollow-core fiber (AR-HCF) to deliver up to 300 W continuous-wave laser power at 1080 nm wavelength from a commercial fiber laser source. A near-diffraction-limited beam is measured at the output of the AR-HCF and no damage to the uncooled AR-HCF is observed for several hours of laser delivery operation. The limit of AR-HCF coupling efficiency and laser-induced thermal effects that were observed in our experiment are also discussed.


Resistance mechanisms to mitochondrial electron transport inhibitors in a field-collected strain of Tetranychus urticae Koch (Acari: Tetranychidae).
A Belgian field strain (MR-VP) of Tetranychus urticae (Koch) (Acari: Tetranychidae) exhibits different levels of resistance to four frequently used METI (mitochondrial electron transport inhibitor)-acaricides, i.e. tebufenpyrad, fenpyroximate, pyridaben and fenazaquin. Resistance factors for these compounds were 184, 1547, 5971 and 35, respectively. A 23.5-fold increase in 7-ethoxy-4-trifluoromethylcoumarin O-deethylation activity suggested that metabolic resistance through elevated levels of cytochrome P450 dependent monooxygenase-activity is a possible resistance mechanism.However, synergism studies with different metabolic inhibitors revealed some contrasting resistance mechanisms between the METI-acaricides. Tebufenpyrad resistance could only be synergized after pre-treatment with the monooxygenase inhibitor piperonyl butoxide (PBO), whereas pyridaben resistance was strongly synergized both by PBO and the esterase inhibitor S,S,S-tributylphosphorotrithioate (DEF). Resistance levels to fenpyroximate could neither be suppressed by PBO nor by DEF. Although METI-acaricides are structurally related, these findings probably reflect a different role of esterases and mono-oxygenases in metabolic detoxification between these compounds. The overall lack of synergism by diethylmaleate (DEM) suggests that glutathione-S-transferases are not an important factor in resistance to METIs.Reciprocal crosses between susceptible females and resistant males showed no maternal effect, and resistance to METI-acaricides was inherited generally as a dominant trait. Backcrosses with F1 females revealed striking differences in the mode of inheritance. Although resistance to fenpyroximate and pyridaben was under monogenic control, resistance to tebufenpyrad was under control of more than one gene.


Induction of ornithine decarboxylase activity by growth and differentiation factors in FRTL-5 cells.
Induction of ornithine decarboxylase has been correlated with the onset of cellular proliferation and cAMP production. Whether the resulting increases in polyamine levels are essential mediators of growth and/or differentiation or are merely incidental remains controversial. We have used FRTL-5 thyroid cells in culture to study the effects of three growth factors on ornithine decarboxylase activity. These factors [TSH, bovine calf serum, and 12-O-tetradecanoylphorbol-13-acetate (TPA)] are thought to act through different intracellular pathways. TSH stimulates cAMP production in thyroid cells, calf serum acts through ill-defined pathways to stimulate growth, and TPA is known to activate protein kinase C. Bovine calf serum and TSH acted synergistically to induce ornithine decarboxylase activity. Activity was maximal when the phosphodiesterase inhibitor, methyl isobutyl xanthine, was included. Individually, neither serum nor TSH was a potent stimulator of the enzyme. Ornithine decarboxylase mRNA was apparent on Northern blots as a doublet following one hour of exposure to these agents. TPA did not stimulate ornithine decarboxylase activity and had an inhibitory effect on enzyme induction by TSH and serum. Difluoromethylornithine, a specific inhibitor of ornithine decarboxylase, inhibited growth induced by both TPA and TSH in putrescine-free medium. This effect was not apparent in medium containing 10(-5) M putrescine. The data indicate that, although intracellular levels of cyclic AMP regulate ornithine decarboxylase activity, a component in serum is necessary for significant induction of this enzyme. Factors stimulating growth by non-cyclic AMP-dependent pathways may act without apparently stimulating this enzyme, although polyamines appear to be essential for their growth stimulatory effects.


Prevalence of Helicobacter pylori infection in disorders of the upper gastrointestinal tract in south India.
Helicobacter pylori (H. pylori) is the most common human infection. Though most individuals are asymptomatic, H. pylori plays a key role in the aetiology of many upper gastrointestinal disorders. The prevalence of duodenal ulcer in south India is high but there are very few reports regarding the prevalence of H. pylori infection in various upper gastrointestinal disorders in south Indians. Therefore, we studied the prevalence of H. pylori infection in upper gastrointestinal disorders in south Indians. Three hundred and thirty-five patients with various upper gastrointestinal disorders were included in the study. Seventy-five patients with no gastrointestinal disease based on symptoms and endoscopy were taken as controls. The H. pylori status was determined by the urease test, serology and histology and the prevalence compared between various upper gastrointestinal disorders, with controls and with one another. The prevalence of H. pylori was high in the patients and controls. Duodenal ulcer patients had a significantly higher prevalence compared to controls (p < 0.001) and those with other upper alimentary disorders. There was no significant difference between patients with other disorders and controls nor between each other (p > 0.05). The prevalence of H. pylori infection is high in south India. It is closely associated with duodenal ulcer. More population-based studies are required to evaluate the relationship of H. pylori with other disorders of the upper gastrointestinal tract.


[Planning and implementation of evaluation studies in child and adolescent psychiatry].
An introduction into planning and implementing evaluation studies in child and adolescent psychiatry is presented. After the presentation of theoretical aspects of evaluation studies, the necessity of these studies is elucidated. Further, a conceptualisation of the dimensions of a treatment programme and some criteria for the selection of outcome criteria are offered. In addition to symptomatology, other criteria such as family functioning and quality of life are proposed in this context. Furthermore, it is recommended to ascertain data already at the time of the patient's registration. The influence of drop-out rate on the study result makes it necessary to closely consider any missing data. These and other problems lead to the conclusion that a sophisticated and complex evaluation methodology is required to assess the performance of a child and adolescent treatment programme validly. Finally, the design of the Hamburg evaluation study is presented, which is suitable for the implementation of some of the mentioned aspects.


Improving the effectiveness of cancer multidisciplinary team meetings: analysis of a national survey of MDT members' opinions about streamlining patient discussions.
Cancer is diagnosed and managed by multidisciplinary teams (MDTs) in the UK and worldwide, these teams meet regularly in MDT meetings (MDMs) to discuss individual patient treatment options. Rising cancer incidence and increasing case complexity have increased pressure on MDMs. Streamlining discussions has been suggested as a way to enhance efficiency and to ensure high-quality discussion of complex cases. Secondary analysis of quantitative and qualitative data from a national survey of 1220 MDT members regarding their views about streamlining MDM discussions. The majority of participants agreed that streamlining discussions may be beneficial although variable interpretations of 'streamlining' were apparent. Agreement levels varied significantly by tumour type and occupational group. The main reason for opposing streamlining were concerns about the possible impact on the quality and safety of patient care. Participants suggested a range of alternative approaches for improving efficiency in MDMs in addition to the use of treatment protocols and pre-MDT meetings. This work complements previous analyses in supporting the development of tumour-specific guidance for streamlining MDM discussions considering a range of approaches. The information provided about the variation in opinions between MDT for different tumour types will inform the development of these guidelines. The evidence for variation in opinions between those in different occupational groups and the reasons underlying these opinions will facilitate their implementation. The impact of any changes in MDM practices on the quality and safety of patient care requires evaluation.


Relationship between bone-prosthesis bonding and load transfer in total hip reconstruction.
The effect of bone-prosthesis bonding on proximal load transfer is investigated using a coupled experimental and finite element analysis on a synthetic femur. Three-dimensional finite element models for an intact femur and a femur implanted with a cementless prosthesis were constructed from the experimental models used, and the proximal femoral strains recorded for two loading conditions approximating a one-legged stance. The approach was used to investigate a press-fitted and a fully bonded bone-prosthesis structure to identify the stem-bone behaviour for both interface conditions and their implications for proximal bone load transfer. Regression slopes close to unity indicated that the finite element predictions were an accurate estimate of the experimental measurements. Physiological surface strains were recorded only when the abductor force was included in the loading. Meanwhile, experimental measurements and numerical predictions showed that a different load transfer pattern is to be expected for normally press-fitted and glued press-fitted stems. The finite element model for the treated femur, modelling both interface conditions correlated very well with the experimental model. These finite element models subsequently modified and used to analyse the effect of different interface conditions predicted a significant increase in the load transfer to the proximal calcar bone when only proximal bonding is achieved. This study suggests that information obtained for the assessment and prediction of total hip arthroplasty longevity by numerical and experimental techniques used together and in parallel is of greater value than either technique used alone. The employment of a femur analogue as featured in this study is also shown to be a suitable alternative to cadaveric specimens in such an analysis.


Glioblastoma multiforme cells: expression of erythropoietin receptor and response to erythropoietin.
Erythropoietin (EPO) is a glycoprotein hormone that is a primary regulator of erythropoiesis. In erythroid cells, EPO binds to its receptor (EPOR) to stimulate growth, prevent apoptosis, and promote differentiation. Both EPO and EPOR have been found in many normal and tumor nonerythroid cell types. EPO has been reported to stimulate proliferation and inhibit apoptosis of cancer cells. In this study, we found that EPOR is expressed in brain tumors, glioma cell lines and explants, as well as, normal brain. EPO slightly stimulated the growth of serum-starved glioma cells. Furthermore, EPO increased the phosphorylation of AKT through the PI3K pathway in the glioma cells. It also increased the phosphorylation of ERK, c-jun, JNK, as well as, the expression of BCL-2 and BCL-xl in these cells. These results suggest that the EPO-EPOR pathway may promote glioma cell survival and could become a therapeutic target in brain tumors.


Disease relevant modifications of the methylome and transcriptome by particulate matter (PM2.5) from biomass combustion.
Exposure to particulate matter (PM) is recognized as a major health hazard, but molecular responses are still insufficiently described. We analyzed the epigenetic impact of ambient PM2.5 from biomass combustion on the methylome of primary human bronchial epithelial BEAS-2B cells using the Illumina HumanMethylation450 BeadChip. The transcriptome was determined by the Affymetrix HG-U133 Plus 2.0 Array. PM2.5 induced genome wide alterations of the DNA methylation pattern, including differentially methylated CpGs in the promoter region associated with CpG islands. Gene ontology analysis revealed that differentially methylated genes were significantly clustered in pathways associated with the extracellular matrix, cellular adhesion, function of GTPases, and responses to extracellular stimuli, or were involved in ion binding and shuttling. Differential methylations also affected tandem repeats. Additionally, 45 different miRNA CpG loci showed differential DNA methylation, most of them proximal to their promoter. These miRNAs are functionally relevant for lung cancer, inflammation, asthma, and other PM-associated diseases. Correlation of the methylome and transcriptome demonstrated a clear bias toward transcriptional activation by hypomethylation. Genes that exhibited both differential methylation and expression were functionally linked to cytokine and immune responses, cellular motility, angiogenesis, inflammation, wound healing, cell growth, differentiation and development, or responses to exogenous matter. Disease ontology of differentially methylated and expressed genes indicated their prominent role in lung cancer and their participation in dominant cancer related signaling pathways. Thus, in lung epithelial cells, PM2.5 alters the methylome of genes and noncoding transcripts or elements that might be relevant for PM- and lung-associated diseases.


SNPdryad: predicting deleterious non-synonymous human SNPs using only orthologous protein sequences.
The recent advances in genome sequencing have revealed an abundance of non-synonymous polymorphisms among human individuals; subsequently, it is of immense interest and importance to predict whether such substitutions are functional neutral or have deleterious effects. The accuracy of such prediction algorithms depends on the quality of the multiple-sequence alignment, which is used to infer how an amino acid substitution is tolerated at a given position. Because of the scarcity of orthologous protein sequences in the past, the existing prediction algorithms all include sequences of protein paralogs in the alignment, which can dilute the conservation signal and affect prediction accuracy. However, we believe that, with the sequencing of a large number of mammalian genomes, it is now feasible to include only protein orthologs in the alignment and improve the prediction performance. We have developed a novel prediction algorithm, named SNPdryad, which only includes protein orthologs in building a multiple sequence alignment. Among many other innovations, SNPdryad uses different conservation scoring schemes and uses Random Forest as a classifier. We have tested SNPdryad on several datasets. We found that SNPdryad consistently outperformed other methods in several performance metrics, which is attributed to the exclusion of paralogous sequence. We have run SNPdryad on the complete human proteome, generating prediction scores for all the possible amino acid substitutions. The algorithm and the prediction results can be accessed from the Web site: http://snps.ccbr.utoronto.ca:8080/SNPdryad/ CONTACT: email@example.com Supplementary information: Supplementary data are available at Bioinformatics online.


Soluble adhesion molecule profiling in preoperative infants with biliary atresia.
Intercellular adhesion molecule (ICAM) expression in liver and bile duct remnant is a feature of the inflammatory component of biliary atresia (BA). Circulating levels of such soluble adhesion molecules (SAM) should reflect intrahepatic disease and would prove a useful adjunct in the evaluation of BA. Serum ICAM-1 (sICAM-1), serum vascular cell adhesion molecule (sVCAM-1), and serum E selectin (sE-selectin) were measured by enzyme-linked immunosorbent assay in infants with BA at the time of portoenterostomy and stratified by outcome. Results are quoted as medians (range). Binary logistic regression analysis and actuarial survival curves were used to investigate the relationship of SAM profile to outcome. Significance was assumed at P values of < or = .05. Sixty-one infants with BA were treated between 1996 and 2002 and at follow-up; 39 of these were alive and jaundice-free (good outcome, n = 39); 21 had been transplanted, and 1 died (poor outcome, n = 22). Preoperative values for sICAM-1 were 1233 (400-2000) ng/mL; sVCAM-1, 1204 (517-1921) ng/mL; and sE-selectin, 71 (26-192) ng/mL. sVCAM-1 (P < .0001) and sICAM-1 (P < .0001) significantly increased compared with normal control infants, although sE-selectin did not (P = .17). There was a significant correlation of age at surgery with sICAM-1 (r = 0.33, P = .01) but not with sVCAM-1 (r = 0.16, P = .23) or sE-selectin (r = 0.05, P = .70). Binary logistic regression analysis showed that the variables, sICAM-1, sE-selectin, or age at surgery, were not significant predictors of outcome, although sVCAM-1 approached significance (P = .069). A cutoff value for sVCAM-1 of 1380 ng/mL was defined by receiver operating characteristic curve analysis, and the cohort of patients with sVCAM-1 (> 1380 ng/mL) showed a significantly worse actuarial survival (P = .05). Levels of sICAM-1 and sVCAM-1 are grossly elevated in BA, whereas sE-selectin levels are normal. Only sVCAM-1 levels have prognostic significance. SAM profiling has the potential to monitor the inflammatory process of BA and may guide more novel forms of pharmacological intervention or immunomodulation.


Near-infrared fundus autoflorescence imaging in solar retinopathy.
Solar retinopathy is a rare clinical entity caused by photochemical damage to the retinal pigment epithelium layer and photoreceptors of the fovea. Here we describe a case of a 33-year-old female patient diagnosed by near-infrared fundus autofluorescence imaging for signs of damage to the melanosomes of the retinal pigment epithelium of the fovea. The patient was advised to discontinue looking at the sun with the naked eye.


The Future Hospital: A Business Architecture View.
The future hospital is a resilient, physical learning facility featuring digital enhancement and leveraging an ecosystem of platforms for the Internet of Things (IoT) and analytics, achieving patient-centric care delivery via multidisciplinary healthcare provider teams coordinated to meet patients' medical, psychological, social and economic needs. It exists in a just ecosystem that assimilates the care spectrum from healthy living, the prevention of disease to acute care and the rehabilitation of patients recuperating from illnesses. It will take some time for these future hospitals to be built or for current hospitals to evolve and/or transform, but efforts to spread wisdom among the stakeholders, healthcare providers and patients must start now. The development of the digital components can also begin today, as can competency building for the healthcare providers who will be staffing these future hospitals, ensuring that they are equipped with competent staff employing patient-centric care processes that cater to patients' current and future needs.


Human injuries and fatalities due to venomous marine snails of the family Conidae.
This paper provides the first compilation in more than 30 years of human injuries and fatalities from envenomation by marine gastropod molluscs of the predominantly tropical family Conidae. It seeks to apply recent advances in knowledge of the physiological effects of conopeptides and molecular genetics to improve our understanding of the human responses to stings by species that normally use their venom peptides to paralyze and overcome prey such as polychaete worms, other gastropod molluscs, and fishes. A database has been constructed for the 139 cases accepted as reliably reporting each human injury. It includes data on the species responsible, the time and place where the stinging occurred and the sting site on the victim's body, the time-course of clinical effects, treatment carried out, if any, and outcome. Members of the hyperdiverse genus Conus caused all the injuries, except for 2 cases involving species from the recently separated genus Conasprella. Death occurred in 36 cases, 57 cases presented with serious symptoms but recovered completely, and in 44 cases victims were only minimally affected. A few cases are listed as tentative because the information in the reports was limited or unverifiable. Many cases have undoubtedly gone unreported and been forgotten. No cases are known for the period between the date of the first reliable report in the 17th century, and the mid-19th century. Knowledge of conopeptide molecular structure and function has recently burgeoned, permitting initial exploration of relationships between the symptoms and outcomes of human injuries and modes of action of these mainly small, very toxic neuroactive peptides. These relationships are reviewed here, especially in regard to the severe and fatal cases, with the aim of making recent knowledge accessible to clinicians and others involved in treating the effects of human stings, which continue to be reported. Conus geographus, a specialized predator of fishes, which it paralyzes with its venom and swallows whole, is the most dangerous species to humans. It accounts for about half of the known human envenomations and almost all the fatalities. Children succumb more often to C. geographus stings than adults and stings by larger snails are lethal more often than stings from smaller snails, regardless of the victim's age. Other piscivorous Conus species have stung humans, but with nonlethal results. A few species that normally prey on other gastropods have also seriously injured humans, but most of the fatalities reported have not been confirmed. Most species of Conidae prey only on marine worms; 18 of these species are known to have stung humans, with generally mild effects. Research on the treatment of Conus stings has lagged behind that on the application of conopeptides in pharmacological research and in the development of new pharmaceuticals. However, improved communication and availability of medical aid in remote tropical areas has likely contributed to reducing the mortality rate during the last half century.


Tannerella forsythia scavenges Fusobacterium nucleatum secreted NOD2 stimulatory molecules to dampen oral epithelial cell inflammatory response.
The oral organism Tannerella forsythia is auxotrophic for peptidoglycan amino sugar N-acetylmuramic acid (MurNAc). It survives in the oral cavity by scavenging MurNAc- and MurNAc-linked peptidoglycan fragments (muropeptides) secreted by co-habiting bacteria such as Fusobacterium nucleatum with which it forms synergistic biofilms. Muropeptides, MurNAc-l-Ala-d-isoGln (MDP, muramyl dipeptide) and d-γ-glutamyl-meso-DAP (iE-DAP dipeptide), are strong immunostimulatory molecules that activate nucleotide oligomerization domain (NOD)-like innate immune receptors and induce the expression of inflammatory cytokines and antimicrobial peptides. In this study, we utilized an in vitro T. forsythia-F. nucleatum co-culture model to determine if T. forsythia can selectively scavenge NOD ligands from the environment and impact NOD-mediated inflammation. The results showed that NOD-stimulatory molecules were secreted by F. nucleatum in the spent culture broth, which subsequently induced cytokine and antimicrobial peptide expression in oral epithelial cells. In the spent broth from T. forsythia-F. nucleatum co-cultures, the NOD-stimulatory activity was significantly reduced. These data indicated that F. nucleatum releases NOD2-stimulatory muropeptides in the environment, and T. forsythia can effectively scavenge the muropeptides released by co-habiting bacteria to dampen NOD-mediated host responses. This proof-of-principle study demonstrated that peptidoglycan scavenging by T. forsythia can impact the innate immunity of oral epithelium by dampening NOD activation.


Mechanics of Brain Tissues Studied by Atomic Force Microscopy: A Perspective.
Tissue morphology and mechanics are crucial to the regulation of organ function. Investigating the exceptionally complex tissue of the brain at the sub-micron scale is challenging due to the complex structure and softness of this tissue, despite the large interest of biologists, medical engineers, biophysicists, and others in this topic. Atomic force microscopy (AFM) both as an imaging and as a mechanical tool provides an excellent opportunity to study soft biological samples such as live brain tissues. Here we review the principles of AFM, the performance of AFM in tissue imaging and mechanical mapping of cells and tissues, and finally opening the prospects and challenges of probing the biophysical properties of brain tissue using AFM.


[Keratoconjunctivitis sicca during folicular conjunctivitis in adult patients with Chlamydia pneumoniae etiology (summarizing twelve years study)].
Following the previous study observing keratoconjunctivitis sicca (KCS) after treated follicular conjunctivitis of chlamydial etiology from the years 1999-2002, at this time the authors would like to point out the possible connection between KCS appearance and Chlamydia pneumoniae infection. In the Department of Ophthalmology, during the period of 12 months (in the years 2009 - 2010), there were treated 11 adult patients (6 women and 5 men) in the age of 28 - 81 years (median, 50 years) with follicular conjunctivitis lasting on average 2.5 years and caused by Chlamydia pneumoniae, and simultaneously were diagnosed the symptoms of KCS, out of the total number of 37 examined patients with similar clinical findings. The diagnosis of chlamydial infection in clinical symptoms of follicular conjunctivitis and simultaneous signs of KCS was established partly according to the serologic positivity of chlamydial genus (anti-LPS) and/or species (anti-Ch. pneumoniae and anti-Ch. trachomatis) IgG, IgM, and IgA antibodies together with simultaneous anti-cHSP60 IgG positivity, further using conjunctival prints examined by means of indirect immunofluorescence method using specific monoclonal antibodies; in conjunctival smears and peripheral leucocytes was tested also the presence of Chlamydia pneumoniae DNA. The main subjective complaints of the patients were burning sensation, sticking, or foreign-body sensation; the itching and lacrimation were less common. Among clinical signs dominated the hyperemia and the edema of the lower fornix; the tortuosity of the vessels and the follicles were not always present. The KCS diagnosis was confirmed by Bengal rose staining. In 10 patients, the positive IgA and/or IgM anti-Ch. pneumoniae antibodies were detected. Simultaneously, the positive anti-cHSP60 IgG were confirmed, out of them medium- and high-positive findings were in 7 patients, and anti-LSP antibodies IgA and/or IgM in 8 of them. Positive smears were found in 6 patients. In 2 persons of this group of treated patients, the general serologic findings were not significant, but the Chlamydia pneumoniae infection was confirmed by Chlamydia pneumoniae DNA positivity in peripheral leucocytes; in these 2 patients, the symptoms lasted for one half of the year. As the side findings were the confirmations of active Chlamydia trachomatis infection in other 4 patients of the whole group. The patients with Chlamydia pneumoniae infection were treated systemically by macrolide antibiotic azithromycine for 12 days. After the treatment, above all else the clinical signs of conjunctival inflammation disappeared, but remained KCS symptoms, which were disappearing only slowly. The optimal supporting therapies were lubricant drops with different concentration of sodium hyaluronide without preservatives. Control microbiological examinations during 6 months after the treatment termination confirmed negative smears findings, but the antibody levels were not decreased significantly. This study confirmed the possible connection between the KCS and Chlamydia pneumoniae infection in simultaneous clinical signs of follicular conjunctivitis. Along with that the study pointed out the necessity of simultaneous evaluating of the microbial findings and clinical symptoms to consider the systemic antibiotic treatment.


Patient preference disability questionnaire in systemic sclerosis: a cross-sectional survey.
To assess patient priorities concerning disability in systemic sclerosis (SSc). A total of 150 SSc patients (22 men) fulfilling the American College of Rheumatology and/or LeRoy and Medsger criteria for SSc were evaluated by the McMaster Toronto Arthritis Patient Preference Disability Questionnaire (MACTAR), Karnofsky performance status (KPS), Cochin Hand Function Scale, Health Assessment Questionnaire (HAQ), Hospital Anxiety and Depression Scale, Mouth Handicap in SSc (MHISS) scale, and global perception regarding their health status. Correlations between scores were analyzed using Spearman's coefficient. Logistic regression analysis was used to determine factors associated with patients' global perception of their health. Of the patients investigated, 81 (54%) had limited cutaneous SSc, 65 (43.3%) diffuse SSc, and 4 (2.7%) limited SSc. The 3 disability domains most often cited were walking (82 patients [54.6%]), housekeeping (67 patients [44.6%]), and sport activities (59 patients [39.3%]). The MACTAR score correlated moderately with KPS (r = 0.58) but only weakly with the HAQ score (r = 0.38). In multivariate analysis, 2 factors were associated with patients' negative global perception of their health status: KPS (odds ratio [OR] 1.07, 95% confidence interval [95% CI] 1.00-1.15) and MHISS score (OR 0.93, 95% CI 0.88-0.99). For assessing SSc patient priorities concerning disability, the MACTAR has acceptable construct validity. Its weak correlation with the HAQ suggests that it adds useful information on disability.


Correlation Between Serum 25(OH)D and Abdominal Visceral Fat Area in Patients with Type 2 Diabetes Mellitus in the Context of different Bone Mass.
To investigate the correlation between serum levels of 25-hydroxy vitamin D [25(OH)D] and the visceral fat area of patients with type 2 diabetes mellitus (T2DM) in the context of different bone mass. A total of 180 patients with T2DM were randomly selected for bone mineral density (BMD) examination. According to the results, they were divided into three groups: T2DM normal bone group (group A); T2DM bone mass reduction group (group B); T2DM osteoporosis group (group C). Serum 25(OH)D levels in NC group, A group, B group and C group decreased in turn, and Visceral fat area (VFA) in group B and group C were significantly higher than those in group A and NC [(29.41±4.87) vs. (22.76±4.23) vs. (17.78±3.61) vs. (9.70±3.01), P<0.05], [(117.76±38.79), (125.08±37.90) vs. (89.79±26.51), (97.53±28.61), P<0.05]. Pearson correlation analysis showed that L1-L4 lumbar vertebrae bone density was positively correlated with 25(OH)D and VFA; left femoral neck bone density was positively correlated with 25(OH)D, and negatively correlated with VFA. Serum 25(OH)D and VFA may be associated with the development of T2DM combined with OP.


Stabilization and manipulation of electronically phase-separated ground states in defective indium atom wires on silicon.
Exploration and manipulation of electronic states in low-dimensional systems are of great importance in the fundamental and practical aspects of nanomaterial and nanotechnology. Here, we demonstrate that the incorporation of vacancy defects into monatomic indium wires on n-type Si(111) can stabilize electronically phase-separated ground states where the insulating 8×2 and metallic 4×1 phases coexist. Furthermore, the areal ratio of the two phases in the phase-separated states can be tuned reversibly by electric field or charge doping, and such tunabilities can be quantitatively captured by first principles-based modeling and simulations. The present results extend the realm of electronic phase separation from strongly correlated d-electron materials typically in bulk form to weakly interacting sp-electron systems in reduced dimensionality.


Plasma concentration of oestradiol-17beta in premature thelarche and in different types of sexual precocity.
Plasma Oe2 concentration was measured by radioimmunoassay in patients with premature thelarche, with precocious puberty and in 29 normal controls. The mean plasma Oe2 was 1.5 pg/ml (0-7.2) in normal prepubertal girls, 23.8 +/- 17.8 (SD) in pubertal girls, 50.2 (+/- 19.4) in the follicular phase, and 94.2 (+/- 19.5) in the luteal phase of normal adult females. Ten girls with premature thelarche had a mean of 7.7 +/- 6.6 pg/ml. Three of them showed higher values than the other 7, suggesting that in these cases, elevated levels of plasma Oe2 might have played a role in the development of breast tissue. Ten untreated girls with idiopathic precocious sexual development had a mean of 51.6 +/- 42.9 pg/ml while 6 patients treated with 150 mg per week of medroxyprogesterone acetate had a mean of 11.4 +/- 2.5 pg/ml. Two patients with Down's syndrome, hypothyroidism and sexual precocity had plasma Oe2 of 144 and 31.5 which fell to 24.7 and 8 pg/ml, respectively, after thyroid replacement. One girl with a granulosa cell tumour had a basal value of 304 pg/ml and a concentration of 27 pg/ml after surgery.


MRI of the hip: current concepts on bone marrow oedema.
Bone marrow oedema (BME) is commonly found in hip MR imaging. BME is a nonspecific finding which can be related to a wide variety of diseases. However, there are cases where its existence or its pattern can narrow the differential or even point to a specific diagnosis. Therefore, deep understanding of its pathophysiology is of utmost importance for clinicians and radiologists in order to avoid diagnostic pitfalls. We hereby present currently established knowledge on BME related to adult hip pathology, aiming to raise physicians' suspicion and to highlight that it is mandatory to combine specific imaging patterns with the history, clinical and laboratory findings in order to achieve accurate diagnosis.


On-site cellulase production by Trichoderma reesei 3EMS35 mutant and same vessel saccharification and fermentation of acid treated wheat straw for ethanol production.
Bioethanol production from lignocellulosic raw materials involves process steps like pre-treatment, enzymatic hydrolysis, fermentation and distillation. In this study, wheat straw was explored as feedstock for on-site cellulase production by T. reesei 3EMS35 mutant, and as a substrate for second generation bioethanol production from baker yeast. Scanning electron microscopy (SEM) and X-ray diffractography (XRD) of untreated wheat straw (UWS) and acid treated wheat straw (TWS) were done to understand the structural organization and changes in the cellulase accessibility and reactivity. The effect of delignification and structural modification for on-site cellulase enzyme production was comparably studied. The efficiency of crude cellulase enzyme for digestion of UWS and TWS and then production of ethanol from TWS was studied using same-vessel saccharification and fermentation (SVSF) technique, both in shaking flasks as well as in fermenters. Two different methods of operation were tested, i.e. the UWSEnz method, where UWS was used for on-site enzyme production, and TWSEnz method where TWS was applied as substrate for cellullase production. Results obtained showed structural modifications in cellulose of TWS due to delignification, removal of wax and change of crystallinity. UWS was better substrate than TWS for cellulase production due to the fact that lignin did not hinder the enzyme production by fungus but acted as a booster. On-site cellulase enzyme produced by T. reesei 3EMS35 mutant hydrolyzed most of cellulose (91 %) in TWS within first 24 hrs. Shake flasks experiments showed that ethanol titers and yields with UWSEnz were 2.9 times higher compared to those obtained with TWSEnz method respectively. Comparatively, titer of ethanol in shake flask experiments was 10 % higher than this obtained in 3 L fermenter with UWSEnz. Outcomes from this investigation clearly demonstrated the potential of on-site cellulase enzyme production and SVSF for ethanol production from wheat straw.


[Anterior resection of the rectum via laparoscopy].
Of the newer laparoscopic procedures for colorectal cancers the abdominoperineal resection of the rectum was previously communicated in this journal. The laparoscopic anterior resection is a more technically demanding operation. We report the case of 63 years old women with a rectal cancer at 9 cm from the anus. The procedure was realised under general anaesthesia, with a urinary catheter in place. An blunt obturator was introduced into the uterine cavity and used to manipulate the uterus during the procedure. After establishing of the pneumoperitoneum, a 30 degrees telescope was introduced through the umbilical port (10 mm). The 30 degrees telescope allows visualization of the splenic flexure and rectal side wall. The main steps of the procedure were the exploration of the abdominal cavity, mobilization of the descendent colon and sigmoid, exposure of the left ureter, division of the left sided peritoneal leaf and division of the inferior mesenteric vessels. The anterior dissection of the rectum was facilitated by manipulation of the uterus. The rectum was then mobilized with the "abdominalization of the tumor". A 5 cm left paramedian vertical incision was performed; through this incision a stapler was introduced and the rectum was transected 5 cm under the tumor. The rectum with the tumor, the sigmoid and mesosigmoid were exteriorized through the incision; the sigmoid was divided with a linear stapler cutter 20 cm above the tumor and the specimen was removed extracorporeally. The anvil of a circular stapler was secured in the left colon and returned in the abdominal cavity. The body of the stapler was inserted into the rectal stump through the anus and connected intracorporeally to the anvil. The postoperative evolution was simple; flatus and feces were passed at 48 hours. The patient was discharged to home after 6 days, 13 months after operation she has no morbidity.


A theoretical model for magneto-acoustic imaging of bioelectric currents.
A theoretical model of magneto-acoustic current imaging is derived, based on fundamental equations of continuum mechanics and electromagnetism. In electrically active tissue, the interaction between an applied magnetic field, B, and action currents, J, creates a pressure distribution. In the near field limit, this pressure obeys Poisson's equation, with a source term (delta x J).B. The displacement and pressure fields are calculated for a dipole (q), oriented either parallel or perpendicular to the applied magnetic field (B), at the center of an elastic, conducting sphere (radius a, shear modulus G). Surface displacements are on the order of qB/(4 pi Ga), which is about 1 nm for typical biological parameters. If the applied magnetic field is changing with time, eddy currents induced in the tissue may be larger than the action currents themselves. The frequency of the pressure and displacement arising from these eddy currents, however, is twice the frequency of the applied magnetic field, so it may be possible to eliminate this artifact by filtering or lock-in techniques. Magneto-acoustic and biomagnetic measurements both image delta x J in a similar way, although magneto-acoustic current imaging has the disadvantage that acoustic properties vary among tissues to a greater degree than do magnetic properties.


New invasive serotype of Escherichia coli.
A new invasive Escherichia coli strain which presented delayed or variable reactions in lactose, mucate, and acetate tests is described. It did not agglutinate with antisera for all known E. coli and Shigella groups A, B, C, D, and provisional Shigella serovar. We propose the designation of E. coli BH until the situation of its 0 antigen is settled.


Metastatic subcutaneous zygomycosis following intravenous and intracerebral inoculation of Absidia corymbifera spores.
A subcutaneous granuloma developed in a C3H mouse ca. 5 months after sequential intravenous and intracerebral inoculation of Absidia corymbifera spores. A. corymbifera was demonstrated microscopically and culturally in the lesion. Infection is believed to have been secondary to fungal infection of the kidney. The histopathological features of the lesion are described.


Chondral resurfacing and high tibial osteotomy in the varus knee.
Treatment of patients with degenerative knees and varus malalignment presents a difficult clinical problem. Combining a medial opening wedge high tibial osteotomy with the microfracture chondral resurfacing procedure is a viable treatment option. Retrospective review of prospectively collected data. A group of 38 consecutive patients (mean age, 51.3 years; range, 34 to 72 years; 29 men and 10 women) with varus malalignment and chondral lesions who were treated with chondral resurfacing (an abrasion and microfracture technique) combined with a medial opening wedge high tibial osteotomy. All patients had >5 degrees of varus malalignment. Patients were evaluated preoperatively with the Lysholm and Western Ontario & McMasters Universities Osteoarthritis Index scoring systems and at a minimum of 2 years follow-up. Thirty-three of 38 patients (87%) were available for 2-year follow-up (average, 45 months; range, 24 to 80 months). Lysholm scores improved from a preoperative score of 43.5 to 78.0 at follow-up; Western Ontario & McMasters Universities Osteoarthritis Index scores improved from 45.8 to 16.2. The average Tegner score was 5.0. Combining a medial opening wedge high tibial osteotomy with the microfracture chondral resurfacing procedure in the varus knee is an effective method of decreasing pain and increasing function at a minimum of 2 years follow-up.


Development of a specific and sensitive method for the detection of glyphosate pesticide and its metabolite in tea using dummy molecularly imprinted solid-phase extraction coupled with liquid chromatography-tandem quadrupole mass spectrometry.
Glyphosate, a widely used herbicide, and its primary metabolite aminomethyl phosphonic acid have been found to cause environmental and ecological issues and threaten human health. The conventional pretreatment method was insufficient for the extraction, concentration, and enrichment of trace substances, resulting in poor specificity. Thus, our objective was to develop a method for glyphosate pesticide detection using dummy molecularly imprinted solid-phase extraction (DMI-SPE) combined with liquid chromatography-tandem quadrupole mass spectrometry (DMI-SPE-LC/MS/MS). The sol-gel method was used to prepare the molecularly imprinted material, using glyphosine as the dummy template molecule, to achieve specific adsorption to glyphosate and reduce costs. The optimized polymerization conditions achieved maximum adsorption of 28.6 µg/mg glyphosate by the molecularly imprinted material. The established DMI-SPE-LC/MS/MS method was used to detect glyphosate and its metabolite (aminomethyl)phosphonic acid in tea. The concentration ranges of glyphosate and (aminomethyl)phosphonic acid (from 0.05 to 4 µg/mL) were linear with correlation coefficients of 0.999 and 0.991, respectively. The recoveries of (aminomethyl)phosphonic acid at three spiked levels ranged from 79.95% to 83.74%, with RSDs between 6.40% and 7.45%, while the recoveries of glyphosate ranged from 98.69% to 106.26%, with RSDs between 0.91% and 1.18%. Our results demonstrate that the developed DMI-SPE-LC/MS/MS method achieves high sensitivity and specific detection of glyphosate and its metabolite (aminomethyl)phosphonic acid in tea matrices.


'Cooking as a biological trait'.
No human foragers have been recorded as living without cooking, and people who choose a 'raw-foodist' life-style experience low energy and impaired reproductive function. This suggests that cooking may be obligatory for humans. The possibility that cooking is obligatory is supported by calculations suggesting that a diet of raw food could not supply sufficient calories for a normal hunter-gatherer lifestyle. In particular, many plant foods are too fiber-rich when raw, while most raw meat appears too tough to allow easy chewing. If cooking is indeed obligatory for humans but not for other apes, this means that human biology must have adapted to the ingestion of cooked food (i.e. food that is tender and low in fiber) in ways that no longer allow efficient processing of raw foods. Cooking has been practiced for ample time to allow the evolution of such adaptations. Digestive adaptations have not been investigated in detail but may include small teeth, small hind-guts, large small intestines, a fast gut passage rate, and possibly reduced ability to detoxify. The adoption of cooking can also be expected to have had far-reaching effects on such aspects of human biology as life-history, social behavior, and evolutionary psychology. Since dietary adaptations are central to understanding species evolution, cooking appears to have been a key feature of the environment of human evolutionary adaptedness. Further investigation is therefore needed of the ways in which human digestive physiology is constrained by the need for food of relatively high caloric density compared to other great apes.


A possible case of Eagle's syndrome from an Italian ossuary (Chiavari, GE).
Eagle's syndrome is a rare condition associated with the temporal styloid process hypertrophy (Eagle, 1937). It is in most cases asymptomatic, but may be accompanied by dysphagia, pharyngeal or head and neck pain due to neurovascular structure compression. The current study aims to present and discuss the etiology of a possible case of Eagle's syndrome from an Italian ossuary. Skull OC 002/08 shows a unilateral hypertrophic styloid process (48 mm long; 5-7 mm thick). Areas of remodel-ed periosteal swelling are visible at the stylohyoid and stylopharyngeus muscles' insertion, along with evidence of healed trauma to both nasal bones and a depressed fracture on the upper left portion of the frontal. Several factors have been proposed as possible causes of styloid elongation, including anatomical variation, aging and trauma. Evidence of unilateral styloid hypertrophy in association with healed cranial trauma in OC 002/08 suggests a traumatic etiology for the condition.


Spread of ISCR1 elements containing blaDHA-₁ and multiple antimicrobial resistance genes leading to increase of flomoxef resistance in extended-spectrum-beta-lactamase-producing Klebsiella pneumoniae.
Increasing resistance to quinolones, aminoglycosides, and/or cephamycins in extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae exacerbates the already limited antibiotic treatment options for infections due to these microbes. In this study, the presence of resistance determinants for these antimicrobial agents was examined by PCR among ESBL-producing Klebsiella pneumoniae (ESBL-KP) isolates that caused bacteremia. Pulsed-field gel electrophoresis was used to differentiate the clonal relationship among the isolates studied. Transferability and the location of the resistance genes were analyzed by conjugation experiments, followed by DNA-DNA hybridization. Among the 94 ESBL-KP isolates studied, 20 isolates of flomoxef-resistant ESBL-KP were identified. They all carried a DHA-1 gene and were genetically diverse. CTX-M genes were found in 18 of the isolates. Among these DHA-1/CTX-M-producing K. pneumoniae isolates, ISCR1 was detected in 13 (72%) isolates, qnr genes (1 qnrA and 17 qnrB genes) were detected in 18 (100%), aac(6')-Ib-cr was detected in 11 (61%), and 16S rRNA methylase (all armA genes) was detected in 14 (78%). Four transconjugants were available for further analysis, and qnrB4, aac(6')-Ib-cr, armA, and bla(DHA-1) were all identified on these self-transferable bla(CTX-M)-carrying plasmids. The genetic environments of ISCR1 associated with armA, bla(DHA-1), and qnrB4 genes in the four transconjugants were identical. Replicon-type analysis revealed a FIIA plasmid among the four self-transferable plasmids, although the other three were nontypeable. The cotransfer of multiple resistance genes with the ISCR1 element-carrying plasmids has a clinical impact and warrants close monitoring and further study.


Interaction of native and mutant MecI repressors with sequences that regulate mecA, the gene encoding penicillin binding protein 2a in methicillin-resistant staphylococci.
Methicillin resistance in staphylococci is mediated by PBP2a, a penicillin binding protein with low affinity for beta-lactam antibiotics. The gene encoding PBP2a, mecA, is transcriptionally regulated in some clinical isolates by mecR1 and mecI, genes divergently transcribed from mecA that encode a signal transducer and repressor, respectively. The biochemical basis of MecI-mediated mecA transcriptional repression was investigated by using purified MecI. In DNase I protection studies, MecI protected a 30-bp palindrome encompassing the predicted mecA -10 and the mecR1 -35 promoter sequences. The larger palindrome contained 15 bp of dyad symmetry within which was a smaller 6-bp palindrome. Electrophoretic mobility shift assays established a requirement for the entire 15-bp half-site for initial repressor binding. Fragments containing the 30-bp palindrome and the entire mecA-mecR1 intergenic region were retarded in gels as multiple discrete bands varying in molecular size, characteristic of cooperative DNA binding. Glutaraldehyde cross-linking confirmed oligomerization of repressor in solution. A naturally occurring MecI mutant (MecI*; D39G) repressed mecA transcription sixfold less well than the wild type in vivo. Although MecI* protected the same target sequences and exhibited similar gel shift patterns to MecI, 5- to 10-fold more protein was required. MecI* exhibited defective oligomerization in solution, suggesting that the MecI amino terminus is important in protein-protein interactions and that protein oligomerization is necessary for optimum repression.


Microevolutionary processes impact macroevolutionary patterns.
Macroevolutionary modeling of species diversification plays important roles in inferring large-scale biodiversity patterns. It allows estimation of speciation and extinction rates and statistically testing their relationships with different ecological factors. However, macroevolutionary patterns are ultimately generated by microevolutionary processes acting at population levels, especially when speciation and extinction are considered protracted instead of point events. Neglecting the connection between micro- and macroevolution may hinder our ability to fully understand the underlying mechanisms that drive the observed patterns. In this simulation study, we used the protracted speciation framework to demonstrate that distinct microevolutionary scenarios can generate very similar biodiversity patterns (e.g., latitudinal diversity gradient). We also showed that current macroevolutionary models may not be able to distinguish these different scenarios. Given the compounded nature of speciation and extinction rates, one needs to be cautious when inferring causal relationships between ecological factors and macroevolutioanry rates. Future studies that incorporate microevolutionary processes into current modeling approaches are in need.