Healthcare providers' views on the acceptability of financial incentives for breastfeeding: a qualitative study.
Despite a gradual increase in breastfeeding rates, overall in the UK there are wide variations, with a trend towards breastfeeding rates at 6-8 weeks remaining below 40% in less affluent areas. While financial incentives have been used with varying success to encourage positive health related behaviour change, there is little research on their use in encouraging breastfeeding. In this paper, we report on healthcare providers' views around whether using financial incentives in areas with low breastfeeding rates would be acceptable in principle. This research was part of a larger project looking at the development and feasibility testing of a financial incentive scheme for breastfeeding in preparation for a cluster randomised controlled trial. Fifty-three healthcare providers were interviewed about their views on financial incentives for breastfeeding. Participants were purposively sampled to include a wide range of experience and roles associated with supporting mothers with infant feeding. Semi-structured individual and group interviews were conducted. Data were analysed thematically drawing on the principles of Framework Analysis. The key theme emerging from healthcare providers' views on the acceptability of financial incentives for breastfeeding was their possible impact on 'facilitating or impeding relationships'. Within this theme several additional aspects were discussed: the mother's relationship with her healthcare provider and services, with her baby and her family, and with the wider community. In addition, a key priority for healthcare providers was that an incentive scheme should not impact negatively on their professional integrity and responsibility towards women. Healthcare providers believe that financial incentives could have both positive and negative impacts on a mother's relationship with her family, baby and healthcare provider. When designing a financial incentive scheme we must take care to minimise the potential negative impacts that have been highlighted, while at the same time recognising the potential positive impacts for women in areas where breastfeeding rates are low.


[Standardizing the diagnosis of acute ischemic stroke--the concept of an institutional manual and its clinical impact].
In the management of acute ischemic stroke, a diagnostic procedure is critically important. However, till date, no guideline or consensus regarding a standard diagnosis procedure has been established. We hereby report an institutional manual for the consistent diagnosis of acute ischemic stroke. The institutional manual was based on the National Institute of Neurological Disorders and Stroke III classification; however, the criteria for each stroke subtype were clearly defined using a rating system that included the representative observations specific to each subtype. The present manual was prepared in order to clearly define the characteristics of the stroke subtypes, which are often ambiguous in the clinical settings, and to design a diagnostic procedure within the institute with more emphasis on standardization rather than achieving complete accuracy. Several characteristic points were considered while doing this: lacunar infarctions should be clearly differentiated from other small infarctions, subtypes of "suspicious cardioembolism," "atherothrombotic infarction (embolic mechanism)," and "infarction due to arterial dissection" should be determined separately in order to identify the causal mechanism to the extent possible. For a final diagnosis, the patients were examined 3 time points during: at admission, during the subacute stage (day 4-7), and at discharge or more than 2 weeks after the onset of stroke. The current version of the manual has been used since 2006 after a transitional phase from 2004 to 2005. Retrospective comparisons on stroke diagnoses and clinical outcomes were performed between the periods during which the present manual was used and those during which it was not. The present manual was retrospectively applied for diagnosing 311 consecutive ischemic stroke patients who were admitted to our institute within 7 days after the onset in 2003; in the case of 85 patients (29%), the diagnosis was different from that made in 2003. Of all the lacunar infections reported in 2003, 17 were diagnosed as other conditions when the manual was used. Similarly, 45% of the cases of atherothrombotic infarction (thrombotic mechanism) and 24% of those of cardiac embolism in 2003 were diagnosed differently when the present manual was applied. The mean modified Rankin scale at discharge was 2.63 +/- 0.07 (mean +/- standard error of the mean) in patients in 2002 and 2003 (n=491), which was significantly different from 2.32 +/- 0.06 in patients in 2006 through March of 2008 (n=903; p=0.01). The present manual appears to be helpful to improve the uniformity of the diagnoses and the clinical results. It may also assist residents and their mentors in the educational field. The manual will require periodical evaluation and version upgrade in order to maintain its efficacy.


Time sequence of oxidative stress in the brain from transgenic mouse models of Alzheimer's disease related to the amyloid-β cascade.
Alzheimer's disease (AD) is a multifactorial disorder characterized by the presence of amyloid plaques and neurofibrillary tangles (NFTs). Rare early-onset forms of AD are associated with autosomal dominant mutations in the amyloid precursor protein gene, presenilin 1 gene, or presenilin 2 gene. The late-onset form of the disease (LOAD) is the most common form. The causes of LOAD are not yet clarified, but several environmental and genetic risk factors have been identified. Numerous studies have highlighted a role for free radical-mediated injury to brain regions of this illness. In addition, studies from mild cognitive impairment patients suggest that oxidative stress is an early event in the pathogenesis of AD. The associations between these markers of free radical damage and the pathogenic cascades involved in AD are complex. Over the past 2 decades, a number of mouse models have been created to recapitulate the major neuropathological hallmarks of AD, namely amyloid plaques and NFTs. These mice recapitulate many, although not all, of the key features of AD. Some strains of transgenic mice develop amyloid plaques, some accumulate NFTs, and some do both. Here we review the evidence for increased free radical-mediated damage to the brain with particular attention to the stage of the disease in various transgenic models of AD related to the amyloid-β cascade.


Thromboelastography in the management of snakebite-induced coagulopathy: a case series and literature review.
: North American crotaline envenomations are a significant source of morbidity annually. Envenomation is marked by a constellation of effects that can include severe tissue damage, systemic effects such as anaphylaxis, and coagulopathy with bleeding. Traditionally, static plasma-derived tests such as protime and PTT are used to assess coagulation status. However, other tests are available that provide a whole blood assessment of all portions of coagulation including initiation, propagation, and clot maintenance. One such analytical test is TEG. Minimal data is available regarding the effect of snake venom on TEG results. We report on three cases of North American crotaline envenomation with subsequent TEG analysis. Our results show that TEG analysis does relate the same information as traditional monitoring parameters. Although only in an isolated case, the TEG analysis potentially revealed a fibrinolytic process prior to the same process being apparent with traditional monitoring. Future research is necessary to identify the role of TEG in crotaline snakebite management.


Is wider surgical margin justified for better clinical outcomes in patients with resectable hepatocellular carcinoma?
Surgical resection for hepatocellular carcinoma (HCC) is regarded as a curable treatment; however, the postoperative recurrence still poses a challenge to surgeons. The effect of surgical margin on long-term outcome is still controversial, although it has been considered as the treatment-related risk factor for recurrence. A precise assessment of the effects of surgical margin on clinical outcome is required to clarify the issue. A retrospective study was conducted on 407 patients with microscopically complete resection of HCC; they were divided into three groups with surgical margin negative by 1-5 mm (Group A, n = 156), 6-10 mm (Group B, n = 109), and wider than 10 mm (Group C, n = 142). The groups were compared for clinicopathologic characteristics, perioperative features, postoperative recurrence, and long-term outcome. The median follow-up time for all patients was 72.97 months. Recurrence rates were similar among these groups. There was no significant difference in the overall 1-, 3-, and 5-year actual survival rates for the groups on the log-rank test (p = 0.073). After controlling the independent risks for disease-free survival, there was also no significant difference in the 1-, 3-, and 5-year disease-free survival rates for the groups (p = 0.354). The patients with wider surgical margin had worse perioperative outcomes; more patients in this group needed blood transfusion (p < 0.001) and more patients suffered from postoperative complications (p = 0.020). They also had higher in-hospital mortality rate than that of other groups (1.41% vs. 0.64%). No superiority was seen in patients with wider surgical margin in either perioperative features or long-term outcome.


Over-the-counter drugs and prescribing in general practice.
Both the government and the pharmaceutical industry are interested in increasing the use of over-the-counter (OTC) medicines. The reaction on the part of general practitioners is more circumspect. To investigate whether fundholding or dispensing status and patient exemption from, or prepayment of prescription charges influence the behaviour of general practitioners with respect to prescribing preparations otherwise available OTC. Regression analysis of data for all 105 Lincolnshire practices for the fiscal year 1993-94, using the number of items prescribed by the practice that were also available OTC as the outcome variable. Comparison of Audit Commission Thematic Analysis of Prescribing (ACTAP) data for fundholders' and non-fundholders' OTC prescribing in the same year. The prescription of medicines otherwise available OTC is less likely when the practice is fundholding and more likely when the practice has dispensing status. Prescription of such medicines also increases as the proportion of patients exempt from, or having prepaid prescription charges increases.


The polysaccharide fucoidin inhibits the antibiotic-induced inflammatory cascade in experimental pneumococcal meningitis.
There is evidence that the treatment of bacterial meningitis with antibiotics liberates harmful bacterial products in the subarachnoid space (SAS). This enhances meningeal inflammation and in particular the recruitment of leukocytes into the cerebrospinal fluid (CSF), which has been shown to be more harmful than beneficial in this disease. In this study, we used a rabbit meningitis model based on intracisternal injection of live Streptococcus pneumoniae. Ampicillin (40 mg/kg of body weight given intravenously [i.v.] 16 h after induction of meningitis) caused a fivefold increase in CSF leukocytes over a 4-h period. Inhibition of leukocyte rolling by treatment with the polysaccharide fucoidin (10 mg/kg, i.v.) prevented the enhanced leukocyte extravasation into the SAS and attenuated the leakage of plasma proteins over the blood-brain barrier. These results suggest that certain polysaccharides that block leukocyte rolling have the potential to reduce leukocyte-dependent central nervous system damage in bacterial meningitis.


Combination of HGF/MET-targeting agents and other therapeutic strategies in cancer.
MET receptor has emerged as a druggable target across several human cancers. Agents targeting MET and its ligand hepatocyte growth factor (HGF) including small molecules such as crizotinib, tivantinib and cabozantinib or antibodies including rilotumumab and onartuzumab have proven their values in different tumors. Recently, capmatinib was approved for treatment of metastatic lung cancer with MET exon 14 skipping. In this review, we critically examine the current evidence on how HGF/MET combination therapies may take advantage of synergistic effects, overcome primary or acquired drug resistance, target tumor microenvironment, modulate drug metabolism or tackle pharmacokinetic issues. Preclinical and clinical studies on the combination of HGF/MET-targeted agents with conventional chemotherapeutics or molecularly targeted treatments (including EGFR, VEGFR, HER2, RAF/MEK, and PI3K/Akt targeting agents) and also the value of biomarkers are examined. Our deeper understanding of molecular mechanisms underlying successful pharmacological combinations is crucial to find the best personalized treatment regimens for cancer patients.


Hylan g-f 20: review of its safety and efficacy in the management of joint pain in osteoarthritis.
Osteoarthritis (OA) is a chronic degenerative joint disease that is a clinically and economically important disease. The increased prevalence of OA with aging, coupled to the demographics of aging populations, make OA a high priority health care problem. Viscosupplementation (VS) is a well-established treatment option in knee OA that is included in the professional guidelines for treatment of this joint disease, and could potentially provide a useful alternative in treating such patients with painful OA. Theoretically VS is an approach that should apply to all synovial joints. The aim of this review is to assess the efficacy and safety of viscosupplementation with Hylan GF-20 (Synvisc(®)) in the management of joint pain in osteoarthritis. THE FOLLOWING DATABASES WERE SEARCHED: Medline, Database of Abstract on Reviews and Effectiveness, Cochrane Database of Systematic Reviews. Furthermore, the lists of references of retrieved publications were manually checked for additional references. The search terms Review, Viscosupplementation, Osteoarthritis, Hyaluronic acid, Hyaluronan, Sodium Hyaluronate, Hylan GF-20, Synvisc, intra-articular injection were used to identify all studies relating to the use of Synvisc(®) viscosupplementation therapy in OA. Hylan GF-20 is a safe and effective treatment for decreasing pain and improving function in patients suffering from knee and hip OA but new evidences are emerging for its use in other joints.


Evidence for a second peptide cleavage in the C-terminal domain of rodent intestinal mucin Muc3.
Rat intestinal mucin Muc3 (rMuc3), like its human homologue (MUC3) and several other membrane mucins, contains a C-terminally located SEA (sea urchin sperm protein, enterokinase and agrin) module, with an intrinsic proteolytic site sequence G downward arrow SIVV (where G downward arrow S is the glycine serine cleavage site). As shown previously [Wang, Khatri and Forstner (2002) Biochem. J. 366, 623-631], expression of the C-terminal domain of rMuc3 in COS-1 cells yields a V5 epitope-tagged N-terminal glycopeptide of 30 kDa and a Myc- and His epitope-tagged C-terminal glycopeptide of 49 kDa. The present study shows that the 49 kDa membrane-anchored fragment undergoes a further cleavage reaction which decreases its size to 30 kDa. Western blotting, pulse-chase metabolic incubations, immunoprecipitation and deglycosylation with N-glycosidase F were used to detect and identify the proteolytic products. Both the first and second cleavages are presumed to facilitate solubilization of Muc3 at the apical surface of enterocytes and/or enhance the potential for Muc3 to participate in ligand-receptor and signal transduction events for enterocyte function in vivo.


Associations of Sedentary and Physically-Active Behaviors With Cognitive-Function Decline in Community-Dwelling Older Adults: Compositional Data Analysis From the NEIGE Study.
Physical activity can help to protect against cognitive decline in older adults. However, little is known about the potential combined relationships of time spent in sedentary behavior (SB), light-intensity physical activity (LPA), and moderate-to-vigorous physical activity (MVPA) with indices of cognitive health. We examined the cross-sectional associations of objectively-determined sedentary and physically-active behaviors with an indicator of cognitive function decline (CFD) in older adults. A randomly-recruited sample of 511 Japanese older adults (47% male; aged 65-84 years) wore a tri-axial accelerometer for 7 consecutive days in 2017. Cognitive function was assessed by interviewers using the Japanese version of Mini-Mental State Examination, with a score of ≤23 indicating CFD. Associations of sedentary and physically-active behaviors with CFD were examined using a compositional logistic regression analysis based on isometric log-ratio transformations of time use, adjusting for potential confounders. Forty one (9.4%) of the participants had an indication of CFD. Activity compositions differed significantly between CFD and normal cognitive function (NCF); the proportion of time spent in MVPA was 39.1% lower, relative to the overall mean composition in those with CFD, and was 5.3% higher in those with NCF. There was a significant beneficial association of having a higher proportion of MVPA relative to other activities with CFD. LPA and SB were not associated with CFD when models were corrected for time spent in all activity behaviors. Larger relative contribution of MVPA was favorably associated with an indicator of CFD in older adults.


A unique fluorescence response of hexaphenylsilole to methyl parathion hydrolase: a new signal generating system for the enzyme label.
Methyl parathion hydrolase (MPH), an enzyme that catalyses the turnover of methyl parathion (MP) to p-nitrophenol (pNP), can be utilized as an enzyme label. In this paper, a unique fluorescence response of 1,1-bis[4-(diethylaminomethyl)phenyl]-2,3,4,5-tetraphenylsilole (A2HPS) to MPH whose gene was obtained from Pseudomonas sp. strain WBC-3 is described. In the absence of MP, A2HPS could only give a small fluorescence response to the enzyme (I/I0 = 1.1). The detection must be performed under low pH conditions, and the influence of BSA and hemoglobin (Hb) was high; upon addition of the enzyme's substrate, 1 × 10-5μg mL-1 or 2.85 × 10-13 M of the MPH could be reported by A2HPS with a higher I/I0 of 1.7. The detection limit was 105 times more sensitive than that given by a spectrophotometric method. In addition, the assay could be performed at a near neutral pH which was more biocompatible, and little influence was observed from BSA and Hb. The light-on response to the MPH was due to the different quenching effect of the MP and pNP on A2HPS and the improvement in the detection selectivity was due to combining the enzyme reaction with the detection. The findings of this work suggested that A2HPS and MP could form a new reporter system for the MPH enzyme label.


[Studies on chemical constituents of the brown alga Dictyopteris divaricata].
To investigate the chemical constituents of the brown alga D. divaricata, and to test cytotoxicities of the purified compounds. Compounds were isolated by normal phase silica gel, Sephadex LH-20 chromatography and reverse phase HPLC techniques. Their structures were elucidated by spectroscopic methods including IR, MS and NMR. Cytotoxicities were tested by MTT method. Eight compounds were isolated from ethanolic extract of the brown alga D. divaricata and their structures were identified as (-)-torreyol (I), 4beta, 5alpha-dihydroxycubenol (II), 3-farnesyl-p-hydroxybenzioc acid (III), chromazonarol (IV), fucosterol (V), phenyl acetylamine (VI), 4-hydroxybenzoic acid (VII) and n-hexadecanoic acid (VIII). Compound II and IV were obtained from this alga for the first time. The others were isolated from the Dictyotaceae algae for the first time. All compounds were inactive (IC50 > 10 microg x mL(-1)) against human tumor cell lines KB, Bel-7402, PC-3M, Ketr 3 and MCF-7.


Methodological quality of multivariate prognostic models for intracranial haemorrhages in intensive care units: a systematic review.
Patients with severe spontaneous intracranial haemorrhages, managed in intensive care units, face ethical issues regarding the difficulty of anticipating their recovery. Prognostic tools help clinicians in counselling patients and relatives and guide therapeutic decisions. We aimed to methodologically assess prognostic tools for functional outcomes in severe spontaneous intracranial haemorrhages. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses recommendations, we conducted a systematic review querying Medline, Embase, Web of Science, and the Cochrane in January 2020. We included development or validation of multivariate prognostic models for severe intracerebral or subarachnoid haemorrhage. We evaluated the articles following the CHecklist for critical Appraisal and data extraction for systematic Reviews of prediction Modelling Studies and Transparent Reporting of multivariable prediction model for Individual Prognosis Or Diagnosis statements to assess the tools' methodological reporting. Of the 6149 references retrieved, we identified 85 articles eligible. We discarded 43 articles due to the absence of prognostic performance or predictor selection. Among the 42 articles included, 22 did not validate models, 6 developed and validated models and 14 only externally validated models. When adding 11 articles comparing developed models to existing ones, 25 articles externally validated models. We identified methodological pitfalls, notably the lack of adequate validations or insufficient performance levels. We finally retained three scores predicting mortality and unfavourable outcomes: the IntraCerebral Haemorrhages (ICH) score and the max-ICH score for intracerebral haemorrhages, the SubArachnoid Haemorrhage International Trialists score for subarachnoid haemorrhages. Although prognostic studies on intracranial haemorrhages abound in the literature, they lack methodological robustness or show incomplete reporting. Rather than developing new scores, future authors should focus on externally validating and updating existing scores with large and recent cohorts.


Additively Manufactured Detection Module with Integrated Tuning Fork for Enhanced Photo-Acoustic Spectroscopy.
Starting from Quartz-Enhanced Photo-Acoustic Spectroscopy (QEPAS), we have explored the potential of a tightly linked method of gas/vapor sensing, from now on referred to as Tuning-Fork-Enhanced Photo-Acoustic Spectroscopy (TFEPAS). TFEPAS utilizes a non-piezoelectric metal or dielectric tuning fork to transduce the photoacoustic excitation and an optical interferometric readout to measure the amplitude of the tuning fork vibration. In particular, we have devised a solution based on Additive Manufacturing (AM) for the Absorption Detection Module (ADM). The novelty of our solution is that the ADM is entirely built monolithically by Micro-Metal Laser Sintering (MMLS) or other AM techniques to achieve easier and more cost-effective customization, extreme miniaturization of internal volumes, automatic alignment of the tuning fork with the acoustic micro-resonators, and operation at high temperature. This paper reports on preliminary experimental results achieved with ammonia at parts-per-million concentration in nitrogen to demonstrate the feasibility of the proposed solution. Prospectively, the proposed TFEPAS solution appears particularly suited for hyphenation to micro-Gas Chromatography and for the analysis of complex solid and liquid traces samples, including compounds with low volatility such as illicit drugs, explosives, and persistent chemical warfare agents.


A clinicopathological analysis of ovarian tumours.
We performed a retrospective analysis of patients with ovarian tumors who were admitted to the Aga Khan University Hospital from January 1985 to December 1989. Sixty one cases were reviewed. Mean age of the whole group was 44 years. Majority of the patients presented with abdominal pain and distention. Most frequent physical finding was a palpable mass on pelvic or abdominal examination. Overall these patients had a higher incidence of breast cancer than expected in the general population. Two-thirds of the tumors were malignant. Comparison of the patients with malignancy against those with benign tumors failed to show any correlation with parity. Majority of the patients with malignant disease were above forty and had ultrasound showing a cystic mass over 10 cms in size. Cancer was mostly epithelial in origin, with widespread disease (stage III or IV) at the time of presentation. Benign tumors, mostly of germ cell type, were predominantly seen in patients under the age of forty with ultrasound showing cystic mass of any size from under 5 cms to over 10 cms.


The association between the 844ins68 polymorphism in the CBS gene and breast cancer.
The cystathionine beta synthase (CBS) gene plays an important role in homocysteine metabolism because it catalyzes the first step of the transsulfuration pathway, during which homocysteine is converted to cystathionine. Polymorphisms of CBS have been associated with cancer. We examined the role of the 844ins68 polymorphism by comparing the genotypes of 371 healthy Mexican women with the genotypes of 323 Mexican women with breast cancer (BC). The observed genotype frequencies for controls and BC patients were 1% and 2% for Ins/Ins, 13% and 26% for W/Ins, and 86% and 72% for W/W, respectively. We found that the odds ratio (OR) was 2.2, with a 95% confidence interval (95% CI) of 1.5-3.3, p = 0.0001. The association was also evident when comparing the distribution of the W/Ins-Ins/Ins genotypes in patients in the following categories: 1) menopause and high γ-glutamyltransferase (GGT) levels (OR of 2.17, 95% CI: 1.17-4.26, p = 0.02), 2) chemotherapy response and high lactate dehydrogenase (LDH) levels (OR 2.2, 95% CI: 1.08-4.4, p = 0.027), 3) chemotherapy response and high GGT levels (OR 2.46, 95% CI: 1.2-4.8, p = 0.007), and 4) body mass index (BMI) and III-IV tumor stage (OR 3.2, 95% CI: 1.2-8.3, p = 0.013). We conclude that the genotypes W/Ins-Ins/Ins of the 844ins68 polymorphism in the CBS gene contribute significantly to BC susceptibility in the analyzed sample from the Mexican population.


Risk of pregnancy-related hypertension within 5 years of exposure to drinking water contaminated with Escherichia coli O157:H7.
The authors evaluated the risk for pregnancy-related hypertension among previously healthy women who conceived within 5 years of exposure to drinking water contaminated with Escherichia coli O157.H7 in Walkerton, Canada (2000). Chronic hypertension was defined as systolic/diastolic blood pressure >/=140/90 mm Hg before 20 weeks gestation; gestational hypertension was defined as new onset systolic/diastolic blood pressure >/=140/90 mm Hg >/=20 weeks gestation. The incidence of hypertension was compared between women who were asymptomatic during the outbreak to those who experienced acute gastroenteritis. Blood pressure data were available for 135 of 148 eligible pregnancies. The adjusted relative risks for chronic and gestational hypertension were 1.5 (95% confidence interval [CI]: 0.3-7.7) and 1.0 (95% CI: 0.4-2.5), respectively. Mean arterial pressure before 20 weeks gestation was 2.7 mm Hg higher in women who had acute gastroenteritis (95% CI: 0.05-5.4). A trend toward higher chronic hypertension and mean arterial pressure in early pregnancy was observed among women who experienced gastroenteritis after exposure to bacterially-contaminated drinking water.


Design, characterization and in vitro evaluation of stavudine-loaded microspheres.
The aim of the present study was to prepare and evaluate microspheres of Eudragit containing an antiviral drug stavudine. Microspheres were prepared by O/O solvent evaporation method using acetone/liquid paraffin system. The prepared microspheres were characterized for their micromeretic properties and entrapment efficiency; as well by Fourier transformed infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD) and scanning electron microscopy (SEM) which revealed the crystalline nature of drug in a final state. The in vitro studies revealed the controlled release of drug from microspheres up to 12 h and the best fit release kinetics was achieved with a Higuchi plot and found to be diffusion controlled. The yields of preparation and entrapment efficiencies were very high with a larger particle size for all the formulations. Mean particle size, entrapment efficiency and production yield were highly influenced by the type of polymer and polymer concentration.


Conducting human challenge studies in LMICs: A survey of researchers and ethics committee members in Thailand.
Questions have been raised over the acceptability of conducting human challenge studies in low and middle income countries (LMICs). Most of these concerns are based on theoretical considerations and there exists little data on the attitudes of stakeholders in these countries. This study examines the view of researchers and REC members in Thailand regarding the design and conduct of challenge studies in the country. A questionnaire was developed based on ethical frameworks for human challenge studies. The target respondents included those who had experience with health-related research at universities, non-university hospitals, and research institutes. A total of 240 respondents completed the on-line survey. In general, the respondents felt that the ethical issues raised by human challenge studies in LMICS do not differ significantly from those in high income countries, including: scientific rationale, safety, appropriate risks, and robust informed consent process. In contrast, issues that have been described as important for human challenge studies in LMICs were rated as having lower importance, including: a publicly available rationale, national priority, and community engagement. Responses did not vary significantly between researchers in different fields, nor between researchers and REC members. These findings provide an important perspective for assessing existing frameworks for human challenges studies in LMICs.


Long-term results of a cementless knee prosthesis with a metal-backed patellar component: clinical and radiological follow-up with histology from retrieved components.
Aseptic loosening of cemented knee arthroplasties has encouraged development of uncemented fixation. The Miller-Galante I (MG I) prosthesis was designed to achieve permanent stability through ingrowth into a titanium fiber mesh. Thirty-five knees in 30 patients with MG I knee replacements have been followed clinically and radiologically with a mean follow-up of 12 years. Twenty knees were revised with a mean follow-up of 5 years. Patellofemoral problems, especially avulsion of the polyethylene from the metal-backed patella and in some cases severe metallosis, have been the main reason for revision. Metal-backed patellar component should be avoided. In addition, instability has been a problem. However, the fixation of the components has been excellent, with a high degree of osseous ingrowth displayed at histological analysis of retrieved components. The clinical and radiological results after revision are in most cases good, despite the femoral component having been left in situ.


Effects of probiotics and antibiotics on the intestinal homeostasis in a computer controlled model of the large intestine.
Antibiotic associated diarrhea and Clostridium difficile infection are frequent complications of broad spectrum antibiotic therapy. Probiotic bacteria are used as therapeutic and preventive agents in these disorders, but the exact functional mechanisms and the mode of action are poorly understood. The effects of clindamycin and the probiotic mixture VSL#3 (containing the 8 bacterial strains Streptococcus thermophilus, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium infantis, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus paracasei and Lactobacillus delbrueckii subsp. Bulgaricus) consecutively or in combination were investigated and compared to controls without therapy using a standardized human fecal microbiota in a computer-controlled in vitro model of large intestine. Microbial metabolites (short chain fatty acids, lactate, branched chain fatty acids, and ammonia) and the intestinal microbiota were analyzed. Compared to controls and combination therapy, short chain fatty acids and lactate, but also ammonia and branched chain fatty acids, were increased under probiotic therapy. The metabolic pattern under combined therapy with antibiotics and probiotics had the most beneficial and consistent effect on intestinal metabolic profiles. The intestinal microbiota showed a decrease in several indigenous bacterial groups under antibiotic therapy, there was no significant recovery of these groups when the antibiotic therapy was followed by administration of probiotics. Simultaneous application of anti- and probiotics had a stabilizing effect on the intestinal microbiota with increased bifidobacteria and lactobacilli. Administration of VSL#3 parallel with the clindamycin therapy had a beneficial and stabilizing effect on the intestinal metabolic homeostasis by decreasing toxic metabolites and protecting the endogenic microbiota from destruction. Probiotics could be a reasonable strategy in prevention of antibiotic associated disturbances of the intestinal homeostasis and disorders.


The minimal instrumentation requirements for Hoechst side population analysis: stem cell analysis on low-cost flow cytometry platforms.
The Hoechst side population (SP) technique is a critical method of identifying stem cells and early progenitors in rodent, nonhuman primate, and human hematopoietic and nonhematopoietic tissues. In this technique, the cell-permeable DNA-binding dye Hoechst 33342 is loaded into the cell population of interest; stem cells and early progenitors subsequently pump this dye out via an ATP-binding cassette membrane pump-dependent mechanism, resulting in a low-fluorescence "tail" (the SP) when the cells are analyzed by flow cytometry. This population contains stem cells and early progenitors. One significant drawback of this method is the requirement of an UV laser to excite the Hoechst 33342. Unfortunately, flow cytometers equipped with UV sources are expensive to own and operate and are not readily available to many laboratories or institutions. In the interests of designing a less expensive flow cytometric system for stem cell analysis, we determined the minimum UV excitation and instrumentation requirements for measuring Hoechst SP. Less than 3 mW of UV laser output was required for adequate resolution of Hoechst SP on two cuvette-based flow cytometers, one of which was a simple, inexpensive benchtop analyzer (the Quanta Analyzer; NPE Systems). Furthermore, Hoechst SP could also be adequately resolved on this epifluorescence-based cytometer platform using two nonlaser UV sources, a mercury arc lamp with a UV bandpass filter and a UV-emitting light-emitting diode. These results suggest that an economical flow cytometric system can be designed that is capable of resolving Hoechst SP, with a cost far lower than most UV laser-equipped commercial systems. An inexpensive system of this type would make Hoechst SP analysis available to a much broader group of stem cell investigators.


Natural killer cell receptors in cattle: a bovine killer cell immunoglobulin-like receptor multigene family contains members with divergent signaling motifs.
Natural killer (NK) cells recognize and kill certain tumor cells, virally infected cells and MHC class I-disparate normal hematopoietic cells. NK cell cytotoxicity is regulated by a multitude of receptors with either activating or inhibitory signaling function. We here report the molecular cloning of bovine CD94 [killer cell lectin-like receptor (KLR)-D1] and NKp46 orthologues, four members of a bovine CD158 [killer cell immunoglobulin-like receptor (KIR)] family, and a novel KLR. This novel receptor was termed KLRJ1 and is most similar to Ly-49 (KLRA). The KLRD1 and KLRJ1 loci were mapped to a bovine NK gene complex on chromosome 5 by radiation hybrid mapping, whereas KIR2DL1 and NKP46 were localized to chromosome 18. Two of the bovine KIR(KIR2DL1 and KIR3DL1) contain immunoreceptor tyrosine-based inhibition motifs (ITIM), suggesting an inhibitory function. Bovine KIR2DS1 and KIR3DS1 lack ITIM but have an arginine-containing motif in their transmembrane domain, similar to primate KIR2DL4. Thus, KIR multigene families with divergent signaling motifs do not only exist in primates. Based on sequence comparison, it appears that the primate and bovine KIR multigene families may have evolved independently.


Gastroprotective effects of pantoprazole against experimental mucosal damage.
The present study investigated the gastroprotective effects of the proton pump inhibitor pantoprazole on gastric mucosal damage induced by ethanol-HCl in rats. Omeprazole was used as reference drug. The morphometric analysis of gastric histological sections revealed that pantoprazole and omeprazole dose-dependently prevented the necrotic mucosal injury evoked by ethanol-HCl (ED50 = 14.1 and 21.6 micromol/kg, respectively). These effects were associated with a marked increment of Alcian blue recovery from gastric bound mucus (ED50 = 18.8 and 29.3 micromol/kg, respectively). In addition, both pantoprazole and omeprazole inhibited gastric acid secretion in pylorus-ligated rats (ED50 = 1.5 and 3.3 micromol/kg, respectively). Further experiments indicated that the protective effects of pantoprazole were not modified by L-365,260 (a gastrin receptor antagonist), suramin (a drug able to interfere with endogenous growth factors), N(G)-nitro-L-arginine (an inhibitor of nitric oxide synthase) or systemic ablation of capsaicin-sensitive sensory nerves, whereas they were partly blocked by indomethacin (an inhibitor of prostaglandin synthesis) and fully prevented by N-ethylmaleimide (a potent blocker of sulfhydryl compounds). The present data provide histomorphometric evidence that: 1) pantoprazole is endowed with gastroprotective properties and is more active than omeprazole in preventing the necrotic mucosal damage induced by ethanol-HCl; 2) according to the rank order of ED50 values, the protective effects of both drugs appear to depend mainly on the enhancement of the gastric mucosal barrier rather than on the inhibition of acid secretion; 3) an increased production of prostaglandins, as well as an increased availability of sulfhydryl radicals at the level of the gastric mucosa may account for the gastroprotective effects of pantoprazole.


Improved electrochemical detection of biogenic amines in Aplysia using base-hydrolyzed cellulose-coated carbon fiber microelectrodes.
A major challenge with electrochemical techniques in vivo, using carbon-fiber microelectrodes, is to achieve sufficient sensitivity to detect the low concentrations of transmitters released by neurons. In particular, when an electrode is inserted into living tissue, its sensitivity is usually substantially decreased as a result of the degradation of the carbon surface by proteins. Here, we show that this decrease can be significantly attenuated by coating the electrode with cellulose acetate. The cellulose film offers a steric barrier that prevents macromolecules from diffusing to the carbon surface and its porosity can be progressively increased by controlled hydrolysis. We compared different cellulose-coated electrodes, either non-hydrolysed or hydrolyzed, in 0.08 N KOH for 10-30 min. We found that dopamine and serotonin detection was blocked by non-hydrolysed cellulose films, but that hydrolysis restored optimal detection similar to uncoated electrodes. Moreover, cellulose films (hydrolyzed for 20 min) significantly diminished electrode degradation in vivo and allowed reliable detection of fast concentration changes with <0.5 s delay, compared to uncoated electrodes. Finally, the sensitivity to endogenous 5-HT release in Aplysia central nervous system was more than doubled with these electrodes. We conclude that the optimal hydrolysis time of cellulose-coated electrodes is approximately 20 min with our protocol and carbon fiber electrodes prepared with this method offer improved sensitivity for the detection of biogenic amines.


Adverse effects of bisphosphonates.
Use of bisphosphonates has been growing steadily in the last decade. This follows the introduction of simpler dosing regimes, the availability of lower-priced generics, and concerns about the safety of hormone-replacement therapy. Bisphosphonates have a relatively good safety record and are tolerated by the majority of patients, but serious adverse events have been recorded in some cases. Only the most common of adverse effects are robustly observable in clinical trials. In general, studies were not powered to detect effects that were lower in incidence than fractures. This review of adverse events in bisphosphonate-treated patients was based on published information from case reports, case series, claims databases, national databases, surveys, adverse event reporting databases, and single or pooled clinical trials. The most common acute adverse events with bisphosphonates for osteoporosis are gastrointestinal discomfort and acute influenza-like illness. Renal complications are very rare with oral bisphosphonates and rare with i.v. bisphosphonates when used appropriately. Based on our current knowledge, skeletal events in the form of osteonecrosis of the jaw and atypical fragility fractures are rare compared with the risk of osteoporotic fractures, at least in patients with the same risk of fractures as those in the phase III trials. It is biologically plausible that atypical fragility fractures could follow from suppression of bone remodeling, but high-quality studies proving causality are lacking. Physicians are advised to critically reassess BMD and risk profile after 3-5 years of therapy to avoid treatment in patients at low risk.


Unconventional Implant Placement IV. Implant Placement through Impacted Teeth to Avoid Invasive Surgery. Long-term Results of 3 Cases.
This paper presents the long-term data of patients that have been treated with an unconventional implant placement protocol to avoid an invasive surgery when edentulism was caused by an impacted tooth. In 2009, the follow-up of this unconven-tional protocol was 2 to 3.5 years; this article documents now the long-term 5- to 8-year follow-up of 3 patients and 5 im-plants. Over this period of time, implant stability was maintained without complications. This unconventional protocol opens intriguing possibilities; however, more patients with long-term follow-up are warranted before endorsing it in routine appli-cation. Nonetheless, it might suggest that there is still room to revisit one of the leading concepts in dental implantology.


[The effect of drug abuse prevention program for elementary school students].
This study is to evaluate the effects of drug abuse prevention program for elementary school students. The design of this study is nonequivalent control group pretest-posttest design. The subjects of experimental group were 27 students and the subjects of control group were 25 students in fifth grade of elementary school in C City, Gyeongsangbuk-do. The experimental group had Drug Abuse Prevention Program, which was two days per week program, for 5 weeks. And post-test was carried out in the same way as the pre-test. Data analysis was done using frequency, percentage, mean, standard deviation, Chi-square test, t-test, Paired Samples t-test using with SPSS WIN 11.0. the experimental group, to which drug abuse prevention program was given, was improved in knowledge of drug and unacceptable attitude of drug compared to the control group, but there were no significant differences of self-esteem and assertiveness between two groups. The drug abuse prevention program was effective to increase knowledge and attitude of drug in elementary school students.


Brain levels of delta1-tetrahydrocannabinol and its metabolites in mice tolerant to the hypothermic effect of delta1-tetrahydrocannabinol.
Repeated voluntary consumption of cannabis in milk (approximately 8 mg per day for 15 days) produced in mice tolerance to the hypothermic effect of tritiated Delta(1)-tetrahydrocannabinol ([(3)H]-Delta(1)-THC; 2 mg/kg i.v.) but not to the effect of [(3)H]-Delta(1)-THC on immobility index. The development of tolerance was not accompanied by any detectable change in levels of radioactivity in the brain assigned to Delta(1)-THC or its metabolites. It was concluded that changes in metabolism or distribution of Delta(1)-THC are not responsible for tolerance at least to the hypothermic effect of Delta(1)-THC in mice.


Evaluation of antimicrobial properties of a novel synthesized nanometric delafossite.
Antibiotics and other antimicrobial compounds are the backbone of clinical medicine. Antimicrobial resistance can cause serious diseases to man. Nanotechnology can improve therapeutic potential of medicinal molecules and related agents. Widespread application of antibiotics and other antimicrobial compounds led to development of multidrug-resistant microbes, so there is need to develop novel therapeutic agents. Novel synthesized nanometric delafossite was assayed against two Gram-positive bacteria (Staphylococcus aureus and Micrococcus luteus), two Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae), four opportunistic fungi (Aspergillus flavus, A. fumigatus, A. niger, and Fusarium solani), and four Candida species (C. albicans, C. parapsilosis, C. krusei, and C. tropicalis) using diffusion assay method. The minimum inhibitory concentration (MIC) of the novel synthesized nanometric delafossite was determined using the dilution method. The assayed compounds showed different degrees of antifungal and antibacterial activities, depending on the annealing temperature of preparation of these compounds. Compounds prepared at room temperature showed greater antimicrobial activities than those prepared at higher temperatures. The antimicrobial activity depends also on the susceptibility of the test microbe.


Numerical matching judgments in children with mathematical learning disabilities.
Both deficits in the innate magnitude representation (i.e. representation deficit hypothesis) and deficits in accessing the magnitude representation from symbols (i.e. access deficit hypotheses) have been proposed to explain mathematical learning disabilities (MLD). Evidence for these hypotheses has mainly been accumulated through the use of numerical magnitude comparison tasks. It has been argued that the comparison distance effect might reflect decision processes on activated magnitude representations rather than number processing per se. One way to avoid such decisional processes confounding the numerical distance effect is by using a numerical matching task, in which children have to indicate whether two dot-arrays or a dot-array and a digit are numerically the same or different. Against this background, we used a numerical matching task to examined the representation deficit and access deficit hypotheses in a group children with MLD and controls matched on age, gender and IQ. The results revealed that children with MLD were slower than controls on the mixed notation trials, whereas no difference was found for the non-symbolic trials. This might be in line with the access deficit hypothesis, showing that children with MLD have difficulties in linking a symbol with its quantity representation. However, further investigation is required to exclude the possibility that children with MLD have a deficit in integrating the information from different input notations.


Are sex and educational level independent predictors of dementia and Alzheimer's disease? Incidence data from the PAQUID project.
To examine the age specific risk of Alzheimer's disease according to sex, and to explore the role of education in a cohort of elderly community residents aged 65 years and older. A community based cohort of elderly people was studied longitudinally for 5 years for the development of dementia. Dementia diagnoses were made according to the DSM III R criteria and Alzheimer's disease was assessed using the NINCDS-ADRDA criteria. Among the 3675 non-demented subjects initially included in the cohort, 2881 participated in the follow up. Hazard ratios of dementia were estimated using a Cox model with delayed entry in which the time scale is the age of the subjects. During the 5 year follow up, 190 incident cases of dementia, including 140 cases of Alzheimer's disease were identified. The incidence rates of Alzheimer's disease were 0.8/100 person-years in men and 1.4/100 person-years in women. However, the incidence was higher in men than in women before the age of 80 and higher in women than in men after this age. A significant interaction between sex and age was found. The hazard ratio of Alzheimer's disease in women compared with men was estimated to be 0.8 at 75 years and 1.7 at 85 years. The risks of dementia and Alzheimer's disease were associated with a lower educational attainment (hazard ratio=1.8, p<0.001). The increased risk of Alzheimer's disease in women was not changed after adjustment for education. Women have a higher risk of developing dementia after the age of 80 than men. Low educational attainment is associated with a higher risk of Alzheimer's disease. However, the increased risk in women is not explained by a lower educational level.


Development of a palliative education assessment tool for medical student education.
Studies assessing palliative care education in U.S. medical schools reveal that little attention is paid to this topic. Although core competencies have been defined, few schools have implemented effective means to incorporate formal palliative care education into undergraduate curricula. To promote reform, each school needs to conduct a thorough assessment to identify palliative care content throughout the four-year curriculum. The authors developed an innovative assessment instrument to facilitate curricular mapping of palliative care education. The Palliative Education Assessment Tool (PEAT) comprises seven palliative care domains: palliative medicine, pain, neuropsychologic symptoms, other symptoms, ethics and the law, patient/family/nonclinical caregiver perspectives on end-of-life care, and clinical communication skills. Each domain details specific curricular objectives of knowledge, skills, and attitudes. Designed as a flexible self-assessment tool, PEAT helps determine the existence of palliative care education, which usually is found in various formats throughout a medical school's curriculum and thus sometimes "hidden." PEAT enables educators to describe a specific, multidimensional aspect of the curriculum and use the information for strategic planning, educational reform, and evaluation. The curricular reform implications of such an instrument are broader than palliative care assessment. A modified version of PEAT can be used to assess systematically other topics that are taught in various formats in the curriculum and to develop collaborative approaches to fulfilling the educational objectives of those topics.


Curvature reduces bending strains in the quokka femur.
This study explores how curvature in the quokka femur may help to reduce bending strain during locomotion. The quokka is a small wallaby, but the curvature of the femur and the muscles active during stance phase are similar to most quadrupedal mammals. Our hypothesis is that the action of hip extensor and ankle plantarflexor muscles during stance phase place cranial bending strains that act to reduce the caudal curvature of the femur. Knee extensors and biarticular muscles that span the femur longitudinally create caudal bending strains in the caudally curved (concave caudal side) bone. These opposing strains can balance each other and result in less strain on the bone. We test this idea by comparing the performance of a normally curved finite element model of the quokka femur to a digitally straightened version of the same bone. The normally curved model is indeed less strained than the straightened version. To further examine the relationship between curvature and the strains in the femoral models, we also tested an extra-curved and a reverse-curved version with the same loads. There appears to be a linear relationship between the curvature and the strains experienced by the models. These results demonstrate that longitudinal curvature in bones may be a manipulable mechanism whereby bone can induce a strain gradient to oppose strains induced by habitual loading.


Audiogenic reflex seizures in cats.
This study aimed to characterise feline audiogenic reflex seizures (FARS). An online questionnaire was developed to capture information from owners with cats suffering from FARS. This was collated with the medical records from the primary veterinarian. Ninety-six cats were included. Myoclonic seizures were one of the cardinal signs of this syndrome (90/96), frequently occurring prior to generalised tonic-clonic seizures (GTCSs) in this population. Other features include a late onset (median 15 years) and absence seizures (6/96), with most seizures triggered by high-frequency sounds amid occasional spontaneous seizures (up to 20%). Half the population (48/96) had hearing impairment or were deaf. One-third of cats (35/96) had concurrent diseases, most likely reflecting the age distribution. Birmans were strongly represented (30/96). Levetiracetam gave good seizure control. The course of the epilepsy was non-progressive in the majority (68/96), with an improvement over time in some (23/96). Only 33/96 and 11/90 owners, respectively, felt the GTCSs and myoclonic seizures affected their cat's quality of life (QoL). Despite this, many owners (50/96) reported a slow decline in their cat's health, becoming less responsive (43/50), not jumping (41/50), becoming uncoordinated or weak in the pelvic limbs (24/50) and exhibiting dramatic weight loss (39/50). These signs were exclusively reported in cats experiencing seizures for >2 years, with 42/50 owners stating these signs affected their cat's QoL. In gathering data on audiogenic seizures in cats, we have identified a new epilepsy syndrome named FARS with a geriatric onset. Further studies are warranted to investigate potential genetic predispositions to this condition.


Differential regulation of matrix metalloproteinases in human cancer cells by exogenous factors.
Matrix degradation by cancer cells is an important attribute of malignant behaviour. We have postulated that during invasion and metastasis, matrix metalloproteinase (MMP) production by cancer cells is modulated by microenvironmental factors. To determine this, we screened human cancer cell lines of diverse histogenesis for their secreted MMP profile following exposure to a diversity of soluble and insoluble agents. Enzymatic changes were observed in the presence of phorbol ester (TPA) and hydrogen peroxide (H2O2). More specifically, we observed: (a) increased secretion and/or activation of gelatinases A (MMP-2) and B (MMP-9) after exposure of 8 cell lines to 10(-6) M TPA; (b) increased activation of interstitial collagenase (MMP-1) caseinolysis after stimulation of 3 cancer cell lines with 10(-7) M TPA; and (c) increased activation of MMP-2 after exposure of 2 cell lines to 0.5 mM H3O2. In functional assays, exposure of lung Calu-6 carcinoma cells to 0.5 mM H2O2 and PC-3 cells to TPA, increased their ability to degrade radiolabelled endothelial cell-derived and osteoblast-like-derived basement membranes, respectively. RNA hybridization studies confirmed and further documented these observations. We conclude that modulation of MMP activity in human cancer cells is a common phenomenon regardless of lineage. Modulatory mechanisms such as increased de novo pro-enzyme synthesis and/or release, and enhanced proenzyme activation may be involved. Furthermore, our results provide evidence that extracellular matrix degradation by human cancer cells during invasion and metastasis may be stimulated by soluble matrix-derived factors in a cell type-specific fashion.


Active force during hypoxia of hypertrophied rabbit right ventricular trabeculae.
Hypertrophy is often accompanied by increased myocardial oxygen demand, but any unique effects of hypoxia on contraction in hypertrophy are unknown. Trabeculae from normal [n = 9; 0.119 +/- 0.014 mm2 (means +/- SE) cross-sectional area] and hypertrophied (pulmonary artery constriction; n = 7; 0.108 +/- 0.028 mm2) rabbit right ventricles were subjected to graded hypoxia (Krebs-Ringer solution, 28 degrees C, 1 Hz stimulus frequency). During normoxia, peak active isometric (Pmax) and resting stress (Prest) at optimum length and peak rate of stress development (dP/dt) in hypertrophy were the same as normal and time to peak stress was longer than normal. Time to peak stress and dP/dt decreased with hypoxia, but time to peak stress remained longer than normal in hypertrophy; Prest was unchanged. The ratio of peak active stress (P) during hypoxia to Pmax decreased linearly with superfusate PO2, but the hypertrophy relationship (y = 4.00 X 10(-3) x + 0.084) is the same as normal (y = 3.70 X 10(-3) x + 0.154; p greater than 0.05). Therefore, a normal level of P was preserved in hypertrophied myocardium and prolonged time to peak stress might have been important for that preservation.


Opioid receptors: some perspectives from early studies of their role in normal physiology, stress responsivity, and in specific addictive diseases.
The early history of research on the possible existence of specific opioid receptors and on developing a new form of pharmacotherapy for the treatment of heroin addiction in New York City, from 1960-1973, along with the special relationships between two leading scientists conducting these research efforts, Dr. Eric Simon and Dr. Vincent P. Dole Jr., are presented in a historical perspective. The linkage of these early efforts and the subsequent identification and the elucidation of the effects of exogenous opiates acting at specific opiate receptors in human physiology, including some findings from perspective studies of heroin addicts at time of entry to and during methadone maintenance treatment, are presented in the context of the important clues which thereby were provided concerning the possible roles of the endogenous opioids in normal mammalian physiology. From many of these early clinical research findings and studies in animal models, the hypothesis that the endogenous opioids system may play an important role in stress responsivity was formulated along with the related hypothesis, first presented in the early 1970s, that an atypical responsivity to stress and stressors might be involved in the acquisition and persistence of, and relapse to specific addictive diseases, including heroin addiction, cocaine dependency and alcoholism. More recent studies of the possible involvement of the specific opioid receptors in these three addictive diseases-heroin addiction, cocaine addiction and alcoholism-from our laboratory are discussed in a historical perspective of the development of these ideas from the early research findings of not only Dr. Eric Simon, but his numerous colleagues in opioid research in the United States and throughout the world.


Characterization of the DNA in Drosophila melanogaster.
DNA has been quantitatively extracted from Drosophila melanogaster at various stages of embryonic development and analyzed by isopycnic centrifugation in CsCl and by fractionation on methylated albumin columns. The DNA is composed of three main classes of DNA, as defined by their buoyant density, rho, in CsCl: a bulk DNA, rho = 1.699 g cm(-3), and two satellite DNAs, rho = 1.685 g cm(-3) and rho = 1.669 g cm(-3). These three types of DNA persist throughout the development of the insect. In the unfertilized egg, 80% of the total DNA consists of the satellite DNAs; this amount decreases to 18% during the first three hours after fertilization and then remains constant through embryogenesis. There is a concomitant increase of the satellite DNA's with the bulk DNA after blastoderm formation.


Vascular Patterns in Retinitis Pigmentosa on Swept-Source Optical Coherence Tomography Angiography.
Retinitis Pigmentosa (RP) represents a retinal dystrophy with an extremely complex pathogenesis further worsened by the impairment of the retinal vascular supply. The main goal of this study was to identify different vascular patterns in RP, by means of optical coherence tomography angiography (OCTA). A total of 32 RP patients (16 males, 50%; mean age 45.93 ± 11.4) and 32 healthy age-matched controls (16 males, 50%; age 42.8 ± 11.2). High resolution OCT and OCTA images were obtained from all participants. Several quantitative parameters were extracted both from structural OCT and OCTA images. A post-hoc analysis assessed the relationship between the quantitative OCTA parameters adopted and the following measures: best corrected visual acuity (BCVA), central macular thickness (CMT) and retinal nerve fiber layer (RNFL). Mean LogMAR BCVA was 0.24 ± 0.32 for RP patients and 0.0 ± 0.0 for controls (p < 0.01). CMT, choroidal thickness and RNFL were statistically different between RP and controls (p < 0.01). OCTA parameters showed strong alterations of the retinal vascular network in RP (all p < 0.01). Several statistically significant correlations were also found. Furthermore, a vessel tortuosity cut-off of 4.80 and a vessel rarefaction cut-off of 0.62 enabled the RP cohort to be divided into two significantly different sub-groups in terms of BCVA, RNFL and CMT. Quantitative OCTA parameters help identify vascular abnormalities in RP, separating two different vascular patterns.


Automated high-content imaging for cellular uptake, from the Schmuck cation to the latest cyclic oligochalcogenides.
Recent progress with chemistry tools to deliver into living cells has seen a shift of attention from counterion-mediated uptake of cell-penetrating peptides (CPPs) and their mimics, particularly the Schmuck cation, toward thiol-mediated uptake with cell-penetrating poly(disulfide)s (CPDs) and cyclic oligochalcogenides (COCs), here exemplified by asparagusic acid. A persistent challenge in this evolution is the simultaneous and quantitative detection of cytosolic delivery and cytotoxicity in a high-throughput format. Here, we show that the combination of the HaloTag-based chloroalkane penetration assay (CAPA) with automated high-content (HC) microscopy can satisfy this need. The automated imaging of thousands of cells per condition in multiwell plates allows us to obtain quantitative data on not only the fluorescence intensity but also on the localization in a very short time. Quantitative and statistically relevant results can be obtained from dose-response curves of the targeted delivery to selected cells and the cytotoxicity in the same experiment, even with poorly optimized cellular systems.


Robust Cooperative Moving Path Following Control for Marine Robotic Vehicles.
This paper presents results on recent developments pertaining to the coordinated motion control of a fleet of marine robotic vehicles. Specifically, we address the Cooperative Moving Path Following (CMPF) motion control problem, that consists of steering the robotic vehicles along a priori specified geometric paths that jointly move according to a target frame, while achieving a pre-defined coordination objective. To this end, each vehicle will need to communicate with their neighbors in order to cooperatively solve the CMPF task. Two distinct robust Moving Path Following motion control strategies for achieving robustness on the moving path following tasks are proposed. Experimental results demonstrating the application of CMPF to marine vehicles in the context of source localization and tracking of underwater targets are presented backed with stability and convergence guarantees.


The development of cutaneous allodynia during a migraine attack clinical evidence for the sequential recruitment of spinal and supraspinal nociceptive neurons in migraine.
Recently, we showed that most migraine patients exhibit cutaneous allodynia inside and outside their pain-referred areas when examined during a fully developed migraine attack. In this report, we studied the way in which cutaneous allodynia develops by measuring the pain thresholds in the head and forearms bilaterally at several time points during a migraine attack in a 42-year-old male. Prior to the headache, he experienced visual, sensory, motor and speech aura. During the headache, he experienced photo-, phono- and odour-phobia, nausea and vomiting, worsening of the headache by coughing or moving his head, and cutaneous pain when shaving, combing his hair or touching his scalp. Comparisons between his pain thresholds in the absence of migraine and at 1, 2 and 4 h after the onset of migraine revealed the following. (i) After 1 h, mechanical and cold allodynia started to develop in the ipsilateral head but not in any other site. (ii) After 2 h, this allodynia increased on the ipsilateral head and spread to the contralateral head and ipsilateral forearm. (iii) After 4 h, heat allodynia was also detected while mechanical and cold allodynia continued to increase. These clinical observations suggest the following sequence of events along the trigeminovascular pain pathway of this patient. (i) A few minutes after the initial activation of his peripheral nociceptors, they became sensitized; this sensitization can mediate the symptoms of intracranial hypersensitivity. (ii) The barrage of impulses that came from the peripheral nociceptors activated second-order neurons and initiated their sensitization; this sensitization can mediate the development of cutaneous allodynia on the ipsilateral head. (iii) The barrage of impulses that came from the sensitized second-order neurons activated and eventually sensitized third-order neurons; this sensitization can mediate the development of cutaneous allodynia on the contralateral head and ipsilateral forearm at the 2-h point, over 1 h after the appearance of allodynia on the ipsilateral head. This interpretation calls for an early use of anti-migraine drugs that target peripheral nociceptors, before the development of central sensitization. If central sensitization develops, the therapeutic rationale is to suppress it. Because currently available drugs that aim to suppress central sensitization are ineffective, this study stresses the need to develop them for the treatment of migraine.


[Mitochondrial 2-methylacetoacetyl-CoA thiolase deficiency in Argentina].
From the description of two pairs of siblings belonging to unrelated families, one Argentine family with a history of consanguinity and Irish ancestry and the other family native of Paraguay, in whom mitochondrial 2-methylacetoacetyl-CoA thiolase deficiency, commonly known as beta-ketothiolase deficiency (beta-KTD, McKusick 203750; EC 188.8.131.52) was recognized. We tried to outline through this experience the clinical and biochemical consequences of this genetic defect in the 6th step of the isoleucine catabolism. The phenotyoic expression presented by the patients belonged to the classical form of beta-KTD. Seven to 15 months was the age at onset of the uniform clinical pattern this being essentially an association of one or several severe ketoacidotic episodes and hyperglycemia which was observed in two patients. The thin-layer chromatography of the tiglylglycine, and dinitrophenylhydrazone of the butanone were positive; aminoacidemia and aminoaciduria revealed normal levels. The organic acids having a unique profile obtained through gaschromatography and mass-spectrometry (GC/MS) showed excretion of large quantities of metabolites characteristic of the disease: 2-methyl-3-hydroxybutirate, 2-methylacetoacetic acid, tiglylglycine and 2-ethylhydracrilic acid which led us to establish the biochemical diagnosis of beta-KTD. The assay of the beta-ketothiolase in lymphocytes and polymorphonuclear leukocytes of the only surviving patient (VT) showed absence of activation by the K+ ion when the acetoacetyl-CoA was used as a substrate. This first Argentine report about beta-KTD leads us to mention three amplifying aspects with regards to previous literature: it adds other different ethnic ancestries of patients, points out a morphological analysis of autopsy material with unchanged structures in the brain, liver and kidneys and marks in the patient VT a dissociation between a symptom-free clinical pattern since age 7 and the persistent biochemical abnormality until the present age, 15 years. The knowledge of the existence of these diseases in our country together with the availability and access to GC/MS of high precision and speed, will allow early diagnosis and better therapeutic results.


U-shape association of serum albumin level and acute kidney injury risk in hospitalized patients.
While an association between hypoalbuminemia and increased risk of acute kidney injury (AKI) is well-established, the risk of AKI development and its severity among patients with elevated serum albumin is unclear. The aim of this study was to evaluate the risk of AKI in hospitalized patients stratified by various admission serum albumin levels. This single-center retrospective study was conducted at a tertiary referral hospital. All adult hospitalized patients who had admission albumin levels available between January 2009 and December 2013 were enrolled. Admission albumin was categorized based on its distribution into six groups (≤2.4, 2.5-2.9, 3.0-3.4, 3.5-3.9, 4.0-4.4, and ≥4.5 mg/dL). The primary outcome was the incidence of hospital-acquired AKI (HAKI). Logistic regression analysis was performed to obtain the odds ratio of AKI for various admission albumin strata using the albumin 3.5 to 3.9 mg/dL (lowest incidence of AKI) as the reference group. Of the total 9,552 studied patients, HAKI occurred in 1,556 (16.3%) patients. The incidence of HAKI among patients with admission albumin ≤2.4, 2.5-2.9, 3.0-3.4, 3.5-3.9, 4.0-4.4, and ≥4.5 mg/dL was 18.3%, 14.3%, 15.5%, 14.2%, 16.7%, and 26.0%, respectively. After adjusting for potential confounders, admission serum albumin levels ≤2.4 and ≥4.5 mg/dL were associated with an increased risk of HAKI with odds ratios of 1.52 (95% CI 1.18-1.94) and 2.16 (95% CI 1.74-2.69), respectively. While stage 1 HAKI was significantly more frequent among patients with admission albumin ≥4.5 mg/dL (23.0% vs. 11.6%, P<0.001), incidence of stage 3 HAKI was higher in those with albumin ≤2.4 mg/dL (2.8% vs 0.3%, P<0.001). Admission serum albumin levels ≤2.4 and ≥4.5 mg/dL were associated with an increased risk for HAKI. Patients with admission albumin ≥4.5 mg/dL had HAKI with a lower intensity when compared with those who had admission albumin levels ≤2.4 mg/dL.


Objective social outcomes index (SIX): a method to summarise objective indicators of social outcomes in mental health care.
No method has yet been established to summarise different objective indicators of social outcomes into one score. The study aimed to develop and test a simple, brief and meaningful index of social outcomes that can be used across a broad range of mental health studies and is potentially obtainable from routine documentation. An index capturing employment, accommodation and living situation (range from 0 to 6; SIX) was devised and tested in three longitudinal datasets of patients with severe mental illness from Germany, Sweden and the UK. SIX showed an almost normal distribution cross-sectionally in all three datasets. Between 58 and 78% of patients changed scores over time. Change scores were weakly associated with change scores of symptoms, quality of life, global functioning and disability. A simple method to summarise different indicators of social outcomes provides meaningful results. It can be widely used in research and routine care.


A Narrative Approach to Synthesizing Research on Vietnamese Bilingual and Monolingual Children.
This review article offers a narrativized synthesis of my research over the past 15+ years with Vietnamese-speaking children based on data collected from individual bilingual and monolingual children in preschool and elementary schools. I begin with a positionality narrative to describe who I am in relation to the research conducted. I provide an overview of the research program including tool building and how my research with bilinguals in the United States led me to international collaborations in Vietnam. I present main findings from this body of work in three areas: typical bilingual development, reading performance in Vietnam, and characteristics of developmental language disorder in the Vietnamese language. Implications within each area are discussed in terms of clinical application and future research directions. Practitioners and researchers alike can freely access the Vietnamese assessment tools created and validated to date from our website, https://vietslp.sdsu.edu/. This research overview aims to offer clinicians and researchers the sociocultural context for understanding the relevance of this body of research. It also serves as an invitation for new generations of scholars, particularly scholars of color, to see their own unique positionings and perspectives as valuable and necessary for scientific innovation and progress. https://doi.org/10.23641/asha.23929491.


Human immune response to group A streptococcal carbohydrate (A-CHO). II. Antigen-independent stimulation of IgM anti-A-CHO production in purified B cells by a monoclonal anti-idiotopic antibody.
The paper describes the induction by a monoclonal anti-idiotopic antibody (anti-Id mAb) of specific antibody production to group A streptococcal carbohydrate (A-CHO) in purified human B cells of several unrelated individuals. The anti-Id mAb, designated 16F498 (anti-Id498), recognizes a recurrent idiotope (Id 498) associated with the combining site of human antibodies to N-acetyl-D-glucosamine (GlcNAc), the immunodominant group of A-CHO. Id498 is expressed on IgM anti-GlcNAc antibodies but does not occur on IgG antibodies with the same specificity. It occurs also on a minor population of IgM antibodies without specificity for A-CHO. Id498 was found in 19 of 27 sera from unselected healthy donors and thus seems to be frequently expressed within the adult B cell repertoire. The in vitro induction of anti-A-CHO antibodies was analyzed in human B cells extensively depleted for T cells. Specific antibody secretion required cross-linked anti-Id which was achieved by coupling the mAb to agarose beads. No antibody secretion could be induced by soluble anti-Id (1 and 10 micrograms/ml). An optimal response required soluble T cell-derived factors which were added as a mixture of recombinant interleukin 2 with a T cell hybridoma supernatant that augments B cell growth and differentiation. Under these conditions an antigen-independent specific increase of IgM anti-A-CHO production (2.6- to 10-fold, or up to 2000 ng/ml respectively) could be induced in blood B cell populations of four of six normal individuals expressing the Id498 at serum level.


Is physical deconditioning a perpetuating factor in chronic fatigue syndrome? A controlled study on maximal exercise performance and relations with fatigue, impairment and physical activity.
Chronic fatigue syndrome (CFS) patients often complain that physical exertion produces an increase of complaints, leading to a greater need for rest and more time spent in bed. It has been suggested that this is due to a bad physical fitness and that physical deconditioning is a perpetuating factor in CFS. Until now, studies on physical deconditioning in CFS have shown inconsistent results. Twenty CFS patients and 20 matched neighbourhood controls performed a maximal exercise test with incremental load. Heart rate, blood pressure, respiratory tidal volume, O2 saturation, O2 consumption, CO2 production, and blood-gas values of arterialized capillary blood were measured. Physical fitness was quantified as the difference between the actual and predicted ratios of maximal workload versus increase of heart rate. Fatigue, impairment and physical activity were assessed to study its relationship with physical fitness. There were no statistically significant differences in physical fitness between CFS patients and their controls. Nine CFS patients had a better fitness than their control. A negative relationship between physical fitness and fatigue was found in both groups. For CFS patients a negative correlation between fitness and impairment and a positive correlation between fitness and physical activity was found as well. Finally, it was found that more CFS patients than controls did not achieve a physiological limitation at maximal exercise. Physical deconditioning does not seem a perpetuating factor in CFS.


Size and shape of airborne asbestos fibres in mines and mills.
There is increasing evidence that fibre size and shape play an important part in the production of health effects related to asbestos. The dimensions of airborne fibres collected at various stages of fibre processing in three mines and six mills producing three types of asbestos were measured using phase contrast light microscopy and transmission electron microscopy. Airborne fibres of different asbestos types had appreciably different size and shape distributions. For a given asbestos type, fibres collected at different stages of processing differed in their size distributions but the differences were considerably less than between fibre types. Most of the airborne fibres to which miners and millers were exposed were short and thin and thus respirable. The physical properties which best differentiated crocidolite fibres from other types of asbestos and which had higher values determined for crocidolite fibres than those obtained for the other types, were median aspect ratio and the proportion of long thin fibres--that is, fibres less than or equal to 0.2 micron in diameter and greater than 5 micron in length as the percentage of total fibres. The median true diameter of amosite fibres was about four and three times higher than the median true diameters of chrysotile and crocidolite fibers respectively. The median true length of amosite fibres was more than four and two times higher than the median true lengths of chrysotile and crocidolite fibres respectively. The observed differences in size and shape of airborne fibres have important implications for the setting of work environmental standards and in explaining differences in health risks associated with different types of fibre.


Suppression of pathogens in properly refrigerated raw milk.
Conflicting claims exist regarding pathogen growth in raw milk. A small pilot study was designed to provide definitive data on trends for pathogen growth and decline in raw bovine milk hygienically produced for direct human consumption. An independent laboratory conducted the study, monitoring growth and decline of pathogens inoculated into raw milk. Raw milk samples were inoculated with foodborne pathogens (Campylobacter, E. coli O157:H7, Listeria monocytogenes, or Salmonella) at lower (<162 colony forming units (CFU) per mL) and higher levels (<8,300 CFU/mL). Samples were stored at 4.4°C and quantified over time after inoculation (days 0, 3, 6, 9, 12, and 14) by standard culture-based methods. Statistical analysis of trends using the Mann-Kendall Trend Test and Analysis of Variance were conducted for 48 time series observations. Evidence of pathogen growth was documented for L. monocytogenes in 8 of 12 replicates (P = 0.001 to P = 0.028). Analysis of variance confirmed significant increases for L. monocytogenes at both initial levels in week 2. No evidence of growth was documented over 14 days for the three pathogens predominantly associated with raw milk outbreaks in the US (Campylobacter, E. coli O157:H7, and Salmonella). Further research is needed to characterize parameters for pathogen growth and decline to support re-assessment of risks that were based on incorrect assumptions about interactions of pathogens with the raw milk microbiota.


Comparative performance of the EQ-5D-5L and SF-6D index scores in adults with type 2 diabetes.
To explore the comparative performance including discriminative and longitudinal validity of EQ-5D-5L and SF-6D index scores in adults with type 2 diabetes. Data from an on-going cohort study of adults with type 2 diabetes in Alberta, Canada, were used. Known-groups approach was used to examine discriminative validity. Correlation and agreement indices and scatter and Bland-Altman plots were used to examine the relationship between the two measures. Longitudinal validity was explored using Wilcoxon signed-rank test, effect size, and standardized response mean. In 1832 participants at baseline (age 64.3, standard deviation 10.6 years; 45% female), median EQ-5D-5L score was 0.85 [interquartile range (IQR) 0.17], and floor and ceiling effects of 0.1 and 16.1%, respectively; median SF-6D score was 0.72 (IQR 0.24), and floor and ceiling effects of 0.1 and 3.2%, respectively. EQ-5D-5L and SF-6D index scores were significantly correlated with an overall Spearman correlation coefficient of 0.73, and an ICC of 0.62 (95% CI 0.42-0.74). Both EQ-5D-5L and SF-6D scores demonstrated statistically significant differences in self-reported chronic conditions, depressive symptoms, and diabetes-related distress, and were able to detect changes in depressive symptoms and diabetes distress across all change groups. Both EQ-5D-5L and SF-6D index scores provide valid measurement in this patient population. Considerable overlap between the measures means it is not necessary to include both in surveys, however, the advantages and disadvantages of each should be considered.


Systematic validation and atomic force microscopy of non-covalent short oligonucleotide barcode microarrays.
Molecular barcode arrays provide a powerful means to analyze cellular phenotypes in parallel through detection of short (20-60 base) unique sequence tags, or "barcodes", associated with each strain or clone in a collection. However, costs of current methods for microarray construction, whether by in situ oligonucleotide synthesis or ex situ coupling of modified oligonucleotides to the slide surface are often prohibitive to large-scale analyses. Here we demonstrate that unmodified 20mer oligonucleotide probes printed on conventional surfaces show comparable hybridization signals to covalently linked 5'-amino-modified probes. As a test case, we undertook systematic cell size analysis of the budding yeast Saccharomyces cerevisiae genome-wide deletion collection by size separation of the deletion pool followed by determination of strain abundance in size fractions by barcode arrays. We demonstrate that the properties of a 13K unique feature spotted 20 mer oligonucleotide barcode microarray compare favorably with an analogous covalently-linked oligonucleotide array. Further, cell size profiles obtained with the size selection/barcode array approach recapitulate previous cell size measurements of individual deletion strains. Finally, through atomic force microscopy (AFM), we characterize the mechanism of hybridization to unmodified barcode probes on the slide surface. These studies push the lower limit of probe size in genome-scale unmodified oligonucleotide microarray construction and demonstrate a versatile, cost-effective and reliable method for molecular barcode analysis.


Polyhydroxyalkanoate quantification in organic wastes and pure cultures using a single-step extraction and 1H NMR analysis.
In this study, a proton nuclear magnetic resonance (1H NMR) method was developed to quantitatively analyze polyhydroxyalkanoate (PHA) content in Cupriavidus necator H16, Azotobacter vinelandii AvOP, and mixed microbial cultures from the effluent of an agricultural waste treatment anaerobic digester. In contrast to previous methods, a single-step PHA extractive method using deuterated chloroform was established, thereby facilitating direct 1H NMR analysis. The accuracy of the method was verified through comparison with well-established gas chromatography (GC) methanolysis techniques. Nile blue fluorescence staining was also carried out to serve as an independent and qualitative indicator of intracellular PHA content. The results indicate that the 1H NMR method is appropriate for rapid and non-destructive quantification of overall PHA content and determination of PHA copolymer composition in a variety of cultures. Notably, this technique was effective in measuring PHA content in full-strength waste samples where high concentrations of background impurities and organic compounds are present. The straightforward procedures minimize error-introducing steps, require less time and materials, and result in an accurate method suitable for routine analyses.


[Content of ceruloplasmin and immunoglobulins of the main classes in the blood sera of patients with herpes virus infection].
In 35 patients with herpes virus infection (males and females aged 25 to 45 years) the content of ceruloplasmin and immunoglobulins of the main classes in the blood sera and the content of IgA in saliva at the stages of exacerbation and remission were evaluated. For control, a group of 35 healthy donors of the same ages were used. In patients with relapsing herpes virus infection even at the period of remission reliably higher levels of ceruloplasmin and immunoglobulins of the main classes were registered in comparison with those in the group of healthy donors. This was indicative of the fact that constant antigenic load caused by virus persistence and, in our opinion, could be regarded as a sign of unfavorable prognosis. At the periods of exacerbation a reliable increased level of secretory IgA was registered in a group of patients with rare relapses of herpes virus infection in comparison with a group of patients with frequent relapses, which showed that patients with rare relapses had a better immune response.


Differences in myocardial alpha- and beta-adrenergic receptor numbers in different species.
In the present study, we tested the hypotheses that 1) different species have different myocardial adrenergic receptor numbers and 2) selected "slow-channel" calcium antagonists compete with alpha-adrenergic antagonists for binding to varying degrees in different species. The data obtained in the present study demonstrate that there is a markedly decreased number of alpha 1-adrenergic and increased number of beta-adrenergic receptors in canine compared with rabbit and rat myocardium. The differences in adrenergic receptor numbers exist without major differences in alpha 1-adrenergic receptor affinity in the species studied. There was no significant difference in left ventricular or plasma catecholamine content between the rat and dog. Selected slow-channel calcium antagonists compete for alpha 1-adrenergic receptor binding in rabbit, rat, and canine myocardium. However, only in rabbit myocardium does verapamil antagonize alpha 1-adrenergic receptor binding at moderate concentrations, whereas verapamil in canine and rat myocardium and 1) 600 in all three species antagonize alpha 1-adrenergic receptor binding only at relatively high concentrations. Nifedipine, a dihydropyridine-type slow-channel calcium antagonist, had no effect on prazosin binding to rat, rabbit, and dog myocardial membranes.


Matrix logarithm parametrizations for neural network covariance models.
Neural networks are commonly used to model conditional probability distributions. The idea is to represent distributional parameters as functions of conditioning events, where the function is determined by the architecture and weights of the network. An issue to be resolved is the link between distributional parameters and network outputs. The latter are unconstrained real numbers whereas distributional parameters may be required to lie in proper subsets, or be mutually constrained, e.g. by the positive definiteness requirement for a covariance matrix. The paper explores the matrix-logarithm parametrization of covariance matrices for multivariate normal distributions. From a Bayesian point of view the choice of parametrization is linked to the choice of prior. This is treated by investigating the invariance of predictive distributions, for the chosen parametrization, with respect to an important class of priors.


Neuropsychiatric Symptoms in Patients With Alzheimer's Disease During SARS-COV-2 Pandemic in Peru.
To evaluate neuropsychiatric symptoms in patients with Alzheimer's disease (AD) and their association with cognition and functionality during lockdown of the COVID-19's first wave. We included 91 patients and caregivers of people with AD from a memory clinic. The RUDAS, M@T, and CDR were administered to patients and NPI/ADCS-ADL to caregivers. Baseline and lockdown measurements scales were analyzed to compare the frequencies at baseline versus lockdown and conditional Odds Ratio (ORc) was calculated for the neuropsychiatric symptoms. During the pandemic, significant increase in the number of cases was observed in depression (23%), agitation (36.8%), aberrant motor activity (12%), sleep disorders (26.3%), and appetite change (12.1%). In worsening of pre-existing symptoms, the most frequent were delusions (75%), followed by sleep disorders (71.7%). Lockdown induces a rapid increase of neuropsychiatric symptoms affecting cognitive symptoms and functionality of Peruvian patients with AD.


Egg production and welfare of laying hens kept in different housing systems (conventional, enriched cage, and free range).
The aim of this study was to compare egg production performance and welfare traits of laying hens kept in conventional cage (CC), enriched cage (EC), and free range (FR). Lohmann Brown laying hens (n = 480 with 160 per housing type) were studied across a production cycle from placement at 17 wk until depopulation at 66 wk. The hens were randomly allocated into cages or pens of housing system groups; within each system there were four replicates with 40 hens in each pen or cage. The hen day egg production (P = 0.037), feed intake (FI) (P < 0.001), egg mass (EM) (P < 0.001), and dirty egg ratio of hens were higher in the FR system but similar in the CC and EC systems. The highest mortality ratio was found in EC system hens (P = 0.020). The best feather score was found in FR system hens (P < 0.001). The worse body wound score was found in EC system hens (P = 0.038). On the other hand, the worse bumble foot and footpad lesions were found in FR system hens (P < 0.001). The highest tibia breaking strength was found in FR system hens compared with in CC and EC system hens (P < 0.001). The highest Heterophil/Lymphocyte ( H/L: ) ratio was found in CC system hens (P = 0.006) but the blood phosphorus ( P: ) level was higher in FR system hens (P = 0.013). The tonic immobility, blood glucose, total cholesterol, triglyceride, and Ca values of hens were found to be similar in all systems (P > 0.05). The hens in the FR system had additional space for optimum comfort and better feather and bone traits, but the dirty egg ratio, feed consumption, and foot lesions were higher than in CC and EC systems.


A chemical approach to myocardial protection and regeneration.
The possibility of generating induced pluripotent stem cells from mouse embryonic fibroblasts and human adult fibroblasts has introduced new perspectives for possible therapeutic strategies to repair damaged hearts. However, obtaining large numbers of adult stem cells is still an ongoing challenge, and the safety of genetic reprogramming with lenti- or retro-viruses has several drawbacks not easy to be addressed. Furthermore, the majority of adult stem cell-based clinical trials for heart regeneration have had generally poor and controversial results. Nonetheless, it is now clear that the injected cells activate the growth and differentiation of progenitor cells that are already present in the heart. This is achieved by the release of signalling factors and/or exosomes carrying them. Along this line, chemistry may play a major role in developing new strategies for activating resident stem cells to regenerate the heart. In particular, this review focuses on small molecule approaches for cell reprogramming, cell differentiation, and activation of cell protection.


Lipids: impact on dietary prescription in diabetes.
The rate of death from coronary artery disease is two to three times higher among individuals with diabetes than among their age- and sex-matched peers. In diabetes, many factors may affect blood lipid levels and complicate formulating an effective dietary prescription. Both Type I and Type II diabetes may have an independent effect on lipids. This article discusses metabolic derivations in carbohydrate and lipid metabolism that contribute to lipid abnormalities. The National Cholesterol Education Program Guidelines, which include dietary recommendations for lowering blood lipid levels by modifying fat intake, are discussed in regard to their effect on blood lipids in diabetes and their effect on other variables, such as blood glucose control. Other treatment approaches common in diabetes and lipid management include low-calorie diets and weight reduction, a change in the fatty acid composition of the diet, and high-carbohydrate diets. An evaluation of the etiologies of lipid abnormalities and interventions directed toward lipid levels and other indexes of diabetes management may lead to improved lipid levels, improved blood glucose control, and avoidance of unnecessary, ineffective, or deleterious dietary prescriptions for individuals with diabetes.


Effect of parathyroid hormone on erythropoiesis.
Inhibitors of erythropoiesis have been found in the blood of uremic patients but their nature has not been identified. These patients have excess blood levels of parathyroid hormone (PTH) and it is possible that PTH inhibits erythropoiesis. The present study was undertaken to examine the effect of intact PTH molecules and some of its fragments on human peripheral blood and mouse bone marrow burst-forming units-erythroid (BFU-E), on mouse bone marrow erythroid colony-forming unit (CFU-E), and granulocyte macrophage progenitors (CFU-GM), and evaluate the interaction between PTH and erythropoietin (Ep) on human BFU-E. Intact PTH (1-84 bPTH) in concentrations (7.5-30 U/ml;) comparable to those found in blood of uremic patients produced marked and significant (P less than 0.01) inhibition of BFU-E and mouse marrow GFU-GM, but not of mouse marrow CFU-E. Inactivation of 1-84 bPTH abolished its action on erythropoiesis. Increasing the concentration of Ep in the media from 0.67 to 1.9 U/ml overcame the inhibitory effect of 1-84 bPTH on BFU-E. The N-terminal fragment of PTH (1-34 bPTH) and 53-84 hPTH had no effect on BFU-E. The results demonstrate that (a) either the intact PTH molecule or a C-terminal fragment(s) bigger than 53-84 moiety exerts the inhibitory effect on erythropoiesis, and (b) adequate amounts of Ep can overcome this action of PTH. The data provide one possible pathway for the participation of excess PTH in the genesis of the anemia of uremia.


Limitations of cyclosporine C2 monitoring in pediatric heart transplant recipients.
Monitoring CSA levels at two h after dosing (C2) has been shown effective in providing adequate CSA-based immunosuppression in clinical trials in adult transplant recipients, but there is limited data regarding C2 monitoring in pediatric transplant recipients. Given the differences in CSA pharmacokinetics between children and adults, a cohort of stable pediatric transplant recipients was converted from monitoring CSA trough (C0) to C2 levels, to establish the clinical utility and safety of C2 monitoring. After an abbreviated AUC(0-5) to establish baseline exposure, subsequent CSA dosing was adjusted based on C2 levels. Additional evaluation included monitoring for rejection, changes in CSA dose, toxicity, serum chemistries, and infection. Twelve heart transplant recipients were enrolled, with mean age 4.8 yr (range: 0.6-14.0). All patients received microemulsified CSA (Neoral((R)); Novartis Pharmaceuticals, East Hanover, NJ, USA) twice daily. Baseline CSA dose was 5.39 +/- 2.05 mg/kg/day (mean +/- s.d.), with mean C0 = 267 +/- 112, C2 = 1065 +/- 565, and AUC(0-5) = 3817 +/- 1435. Only seven participants showed clear CSA peak levels at two h, with five exhibiting delayed peaks at three to five h post-dose. These seven participants completed 48 wk of study, with mean CSA dose decreasing to 4.55 +/- 3.61 mg/kg/day, maintaining mean C2 599 +/- 211 (vs. target C2 = 800). No significant change in serum creatinine was observed, although GFR increased from 76.9 to 107.6 mL/min/1.73 m(2) (p = 0.11). Five patients failed to achieve target C2 levels (>800) during the first four wk, despite comparable AUC values, and were maintained on trough monitoring (C0). Mean systolic and diastolic blood pressures fell slightly, three minor infections were noted during the study period, and one episode of acute rejection occurred, despite stable CSA dosing. Nearly 50% of stable pediatric transplant recipients failed to achieve adequate peak C2 CSA levels during conversion from C0 to C2 monitoring. Age-dependent differences in CSA absorption and/or clearance pharmacokinetics may explain these findings.