A Poly(Propyleneimine) Dendrimer-Based Polyplex-System for Single-Chain Antibody-Mediated Targeted Delivery and Cellular Uptake of SiRNA.
Therapeutics based on small interfering RNAs (siRNAs) offer a great potential to treat so far incurable diseases or metastatic cancer. However, the broad application of siRNAs using various nonviral carrier systems is hampered by unspecific toxic side effects, poor pharmacokinetics due to unwanted delivery of siRNA-loaded nanoparticles into nontarget organs, or rapid renal excretion. In order to overcome these obstacles, several targeting strategies using chemically linked antibodies and ligands have emerged. This study reports a new modular polyplex carrier system for targeted delivery of siRNA, which is based on transfection-disabled maltose-modified poly(propyleneimine)-dendrimers (mal-PPI) bioconjugated to single chain fragment variables (scFvs). To achieve targeted delivery into tumor cells expressing the epidermal growth factor receptor variant III (EGFRvIII), monobiotinylated anti-EGFRvIII scFv fused to a Propionibacterium shermanii transcarboxylase-derived biotinylation acceptor (P-BAP) is bioconjugated to mal-PPI through a novel coupling strategy solely based on biotin-neutravidin bridging. In contrast to polyplexes containing an unspecific control scFv-P-BAP, the generated EGFRvIII-specific polyplexes are able to exclusively deliver siRNA to tumor cells and tumors by receptor-mediated endocytosis. These results suggest that receptor-mediated uptake of otherwise noninternalized mal-PPI-based polyplexes is a promising avenue to improve siRNA therapy of cancer, and introduce a novel strategy for modular bioconjugation of protein ligands to nanoparticles.


Short Communication:Outcomes of Dual Versus Triple Antiretroviral Drug Regimens Among Virally Suppressed Adults in the DC Cohort.
This study explored virological outcomes of two-drug (2DRs) and three-drug (3DRs) antiretroviral regimens in adults with HIV in the DC Cohort. We analyzed 310 treatment-experienced adults with sustained HIV RNA ≤50 copies/mL at baseline, 53 of whom switched to 2DRs and 257 continued 3DRs. Adults on 2DRs and 3DRs had similar demographics (median age 53.3 years, 76.8% cisgender male, 76.1% Black). Adults on 2DRs had more participants with ≥2 comorbidities (62.3% vs. 42.8%, p = .019), had a longer time since HIV diagnosis (median years 20.4 vs. 13.2, p = .017), and received the regimen of interest for a shorter duration (median years 1.3 vs. 3.3, p < .001) compared with adults on 3DRs. Adults receiving 2DRs had a higher, although nonsignificant, risk for virological failure (two consecutive HIV RNA ≥50 copies/mL) at 24 months follow-up than adults on 3DRs (6.7% vs. 1.7%, respectively; p = .10). Future analysis of the effectiveness of 2DRs is needed.


Hair mercury levels of residents in China, Indonesia, and Japan.
The authors used gold-amalgamation cold-vapor atomic absorption spectrometry and ECD-gas chromatography to analyze total mercury and methylmercury levels in hair samples obtained from 362 residents in Harbin, China; Medan, Indonesia; and Tokushima, Japan. In this study, the authors initially questioned whether mercury levels in hair differed among different study areas, and if there were differences, they questioned the contributing factors. In the three countries surveyed, total mercury and methylmercury levels in hair were lowest in residents of China and were highest in residents of Japan. In the district of Tokushima, Japan, total mercury and methylmercury levels were highest in the coastal district, followed by the middle district; the lowest levels occurred in the mountainous district. In Japan, an individual's total mercury level correlated very closely with that person's methylmercury level; in China and Indonesia, the correlation between these 2 parameters was low. No subjects in China or Indonesia had high levels of methylmercury in hair; this was true even if their total mercury levels were high. This finding suggests that the high total mercury levels observed in some residents of China and Indonesia reflected exposure to inorganic mercury. In Japan, mercury (especially methylmercury) levels in hair samples were quite high. Fish and shellfish, caught in seas uncontaminated by human activity, appeared to be major sources of the high levels of hair mercury in Japanese subjects.


Perinatal salivary gland tumours (embryomas).
Epithelial salivary gland tumours presenting in the first month of life (perinatal tumours) are exclusively of major salivary gland origin and predominantly of the parotid gland. Approximately 25 per cent of these rare tumours have been histologically and/or biologically malignant. Except for an occasional pleomorphic and monomorphic adenoma, the majority of the tumours are proliferations suggesting a tumourous arrest of maturation of the salivary gland anlage, hence their designation as embryomas. A case report of an embryoma of the parotid is presented, accompanied by a review of the literature.


Neurodevelopmental outcome in survivors of periventricular hemorrhagic infarction.
Periventricular hemorrhagic infarction is a serious complication of germinal matrix-intraventricular hemorrhage in premature infants. Our objective was to determine the neurodevelopmental and adaptive outcomes of periventricular hemorrhagic infarction survivors and identify early cranial ultrasound predictors of adverse outcome. We retrospectively evaluated all cranial ultrasounds of 30 premature infants with periventricular hemorrhagic infarction and assigned a cranial ultrasound-based periventricular hemorrhagic infarction severity score (range: 0-3) on the basis of whether periventricular hemorrhagic infarction (1) involved > or = 2 territories, (2) was bilateral, or (3) caused midline shift. We then performed neuromotor, visual function, and developmental evaluations (Mullen Scales of Early Learning, Vineland Adaptive Behavior Scale). Developmental scores below 2 SD from the mean were defined as abnormal. Median adjusted age at evaluation was 30 months (range: 12-66 months). Eighteen subjects (60%) had abnormal muscle tone, and 7 (26%) had visual field defects. Developmental delays involved gross motor (22 [73%]), fine motor (17 [59%]), visual receptive (13 [46%]), expressive language (11 [38%]), and cognitive (14 [50%]) domains. Impairment in daily living and socialization was documented in 10 (33%) and 6 (20%) infants, respectively. Higher cranial ultrasound-based periventricular hemorrhagic infarction severity scores predicted microcephaly and abnormalities in gross motor, visual receptive, and cognitive function. In the current era, two thirds of periventricular hemorrhagic infarction survivors develop significant cognitive and/or motor abnormalities, whereas adaptive skills are relatively spared. Higher cranial ultrasound-based periventricular hemorrhagic infarction severity scores predict worse outcome in several modalities and may prove to be a valuable tool for prognostication.


Longitudinal association of glycemia and microalbuminuria: the Framingham Offspring Study.
To assess current and long-term associations of glycemia with microalbuminuria, a marker of generalized endothelial injury. We measured clinical characteristics, fasting plasma glucose, and the urinary albumin-to-creatinine ratio (UACR) in 1,311 men and 1,518 women attending the sixth examination cycle (1995-1998) of the Framingham Offspring Study. After excluding participants with diabetes or cardiovascular disease (CVD) at the baseline examination (1971-1974), we used fasting glucose measured at baseline, examination 6, and at least two additional examinations from 1974 to 1995 in regression models to predict risk for microalbuminuria (UACR > or = 30 mg/g) associated with baseline, current, and 24-year time-integrated glycemia. Microalbuminuria was present in 9.5% of men and 13.4% of women. Among men, age-adjusted odds ratios (95% CI) for microalbuminuria associated with each 0.28 mmol/l (5 mg/dl) increase in baseline, current, and time-integrated glucose levels were 1.12 (1.00-1.16), 1.08 (1.05-1.10), and 1.16 (1.11-1.21), respectively. These effects persisted after adjustment for systolic blood pressure and other confounders. Higher glucose levels also predicted incident diabetes and CVD. Mean time-integrated glucose levels were highest among men who developed both CVD and microalbuminuria (SE 6.82 +/- 0.16 mmol/l), intermediate among men with either condition (6.03 +/- 0.65 mmol/l), and lowest among men with neither condition (5.49 +/- 0.02 mmol/l; P < 0.001 for all pairwise comparisons). We observed similar associations in women. Long-term hyperglycemia and subdiabetic glycemia increase risk for microalbuminuria. Microalbuminuria, type 2 diabetes, and CVD seem to arise together over the course of decades, consistent with the hypothesis that they share a common antecedent.


Long-term culture of porcine bladder epithelial cells.
Epithelial cells from normal pig bladders proliferated when cocultured with lethally irradiated feeder cells of the LA7 rat mammary tumor line. When the bladder cells and feeders were plated together at a confluent density, the bladder cells proliferated as the feeder cells died, resulting in a confluent culture of bladder cells. The bladder cells were successfully subcultured by plating with freshly irradiated LA7 feeder cells. In this way, bladder cells from five pigs were carried to confluency in passages 1, 4, 7, 7, and 13, amounting to at least 6, 18, 24, 26, and 45 doublings in culture, respectively, and none showed signs of slowed proliferation at the time of culture termination. Fibroblasts never became a prominent feature of these cultures, and their frequency was determined to be about 26 fibroblasts per 10(5) cells in passage 9. Pig bladder cells in 0.5% serum doubled in number in slightly over 3 d, whereas cells in 5.0% serum doubled in about 6 d. In fresh medium without feeder cells only minimal proliferation of bladder cells occurred. In LA7-conditioned medium the bladder cell numbers decreased, leading to the conclusion that the stimulus from LA7 cells is mechanically or physically transmitted. The bladder cells reacted with antibodies to keratins 7 and 18 but not to keratin 14 or vimentin. Tight junctions, visualized with an antibody to the ZO1 protein, connected all the cells to their neighbors. Most cells in passage 9 carried the diploid chromosome number of 38.


Obesity and heredity in the etiology of non-insulin-dependent diabetes mellitus in 32,662 adult white women.
Obesity and family history of non-insulin-dependent diabetes mellitus were examined in a cross-sectional study of 32,662 white women in the United States and Canada who were members of the TOPS (Take Off Pounds Sensibly) Club, Inc., with respect to the association between these two factors and the prevalence of diabetes by using questionnaire data originally collected in 1969. A family history index based upon the reported presence of diabetes among siblings, parents, and grandparents was used to grade hereditary factors on a scale of zero to five. The ratio of actual to ideal weight was used to categorize levels of obesity. The prevalence of diabetes increased consistently with increasing levels of either obesity or family history. The two factors appear to present independent risks for diabetes. Among women with no family history of diabetes, the odds ratio for diabetes associated with a weight 75 per cent or more above ideal in comparison with a weight less than 10 per cent above ideal was 7.2. Among women who were less than 10 per cent above ideal weight for their height, a comparison between women with a family history index of 1.5 or greater and women with no family history of the disease showed an odds ratio for diabetes of 6.0. Women with both risk factors (family history index of 1.5 or greater and relative weight of 1.75 or greater) had an overall odds ratio for diabetes of 22.8 when compared with women who had no family history of diabetes and a weight less than 10 per cent above ideal. Further analysis showed the combination of these factors to have a positive synergistic effect. The interaction follows a multiplicative pattern.


Duplex-Specific Nuclease-Mediated Amplification Strategy for Mass Spectrometry Quantification of MiRNA-200c in Breast Cancer Stem Cells.
MicroRNAs (miRNAs) play a significant role in numerous biological processes and are implicated in a range of cancers, including breast cancer. MiRNAs have the potential to be biomarkers in clinical practice because of their distorted and unique expression, especially with regard to their presence in cancer stem cells (CSCs) that have applications in cancer diagnosis and treatment. Thus, the absolute determination of miRNA expression levels is a prerequisite for exploring their applications. Nevertheless, currently available methods may not be adequate for the detection of miRNAs in CSCs due to the inherently low population of these cells. Therefore, we combined a duplex-specific nuclease (DSN)-mediated amplification strategy with liquid chromatography-tandem mass spectrometry (LC-MS/MS) for use in this study. We designed the substrate peptide GDKAVLGVDPFR, which contains the reporter peptide AVLGVDPFR and a tryptic cleavage site (lysine at position 3) in combination with a biotinylated DNA sequence that was complementary to a target miRNA (i.e., miR-200c). Then, this newly synthesized DNA-peptide probe was hybridized with the target miRNA. Upon the introduction of the DSN, the enzyme degraded the probe into two parts where the target miRNA was integrated and thereby triggered further cleavage. The accumulated peptide fragments were ultimately digested with trypsin to release the reporter peptide for LC-MS/MS quantification. Under these circumstances, the miRNA signal was converted and amplified into a mass response of the reporter peptide. After optimization of the parameters, including temperature, hybridization/DSN time, and the amounts of DSN and streptavidin agarose beads, we demonstrated a linear detection range between 1 fM and 200 fM for miR-200c. The detection limit obtained was 3 orders of magnitude lower than those previously reported. Finally, quantification of miR-200c in breast cancer stem cells (BCSCs) and in stem cells isolated from breast tumors was performed. We also compared these data with quantitative reverse transcription PCR (qRT-PCR) results.


Significance of daily rhythm for the interpretation of functional diagnostic procedures.
For the first time, the reaction of the cardiovascular system during day and night in young people was examined. We were mostly interested in the influence of the nervous system and any autonomic mechanism. We examined 20 subjects at 4-hour intervals on the ergometer (active exercise) and on the tilt-table (passive exercise) to find out the rhythm in the circulatory regulation. The test person was kept in vertical position on the tilt-table for 10 minutes. On the ergometer he had to perform a work-load of 100 Watt for 10 minutes. We measured pulse rate and blood pressure; we introduced new terms for the evaluation. "Orthostatic Index" and "Performance Index" will be further explained. The results were: A minimum in the orthostatic stability around 1 p.m. and 1--5 am, and a performance maximum at 1 p.m. and 5 p.m. Further details will be reported.


Novel radioimmunopharmaceuticals for cancer imaging and therapy.
As interest in targeted therapies for cancer increases, radionuclides stand out not only for their ability to be detected by external scintigraphy, but also for their therapeutic capacity. Radionuclides need to be directed to the invading cancer however, and in this context antibodies have been at the forefront of targeting radionuclides for more than 50 years. Antibodies have been dissected and reconstituted in many different forms to fit a multitude of functions. Whether directly coupled to a radionuclide or used in a pretargeting setting, radioimmunoconjugates are being reinvented to meet the many challenges facing cancer detection and therapy today and in the future.


Squamous cell carcinoma of the right kidney on FDG PET-CT.
We report the PET-CT appearance of a newly diagnosed, biopsy proven squamous cell carcinoma of the right kidney in a 61-year-old man who presented with hematuria. This is a rare tumor, with few cases reported in the literature. The patient subsequently underwent right nephrectomy shortly after the initial staging PET-CT study.


Characterization of the Kaposi's sarcoma-associated herpesvirus K1 signalosome.
Kaposi's sarcoma (KS) is a multifocal angiogenic tumor and appears to be a hyperplastic disorder caused, in part, by local production of inflammatory cytokines. The K1 lymphocyte receptor-like protein of KS-associated herpesvirus (KSHV) efficiently transduces extracellular signals to elicit cellular activation events through its cytoplasmic immunoreceptor tyrosine-based activation motif (ITAM). To further delineate K1-mediated signal transduction, we purified K1 signaling complexes and identified its cellular components. Upon stimulation, the K1 ITAM was efficiently tyrosine phosphorylated and subsequently interacted with cellular Src homology 2 (SH2)-containing signaling proteins Lyn, Syk, p85, PLCgamma2, RasGAP, Vav, SH2 domain-containing protein tyrosine phosphatase 1/2, and Grab2 through its phosphorylated tyrosine residues. Mutational analysis demonstrated that each tyrosine residue of K1 ITAM contributed to the interactions with cellular signaling proteins in distinctive ways. Consequently, these interactions led to the marked augmentation of cellular signal transduction activity, evidenced by the increase of cellular tyrosine phosphorylation and intracellular calcium mobilization, the activation of NF-AT and AP-1 transcription factor activities, and the production of inflammatory cytokines. These results demonstrate that KSHV K1 effectively recruits a set of cellular SH2-containing signaling molecules to form the K1 signalosome, which elicits downstream signal transduction and induces inflammatory cytokine production.


Malaria outbreak along the India-Bhutan border town of Saralpara, Assam, India.
A remarkable progress is being made in the South-East Asia region in reducing the burden of malaria. While all countries are committed to malaria elimination, continued transmission at and across international borders poses a threat to achieving this goal. We investigated an outbreak of malaria in the India-Bhutan border area in a district of Assam state, India. A line listing of cases occurring during the outbreak period was made and a house-to-house survey was carried out to identify additional cases in the community. Entomological investigation consisted of light trap collection of vectors. Patients and health workers were interviewed to obtain information on the possible mode of transmission and ongoing inter-country collaboration across the border. Between October 21 to November 18 2022, 37 cases of malaria (all Plasmodium vivax), were diagnosed in Saralpara area of Kokrajhar district of Assam, bordering Bhutan. Of these four were diagnosed in Sarpang hospital, Bhutan. Median age was 20 years (range 3 to 70 years). Male to female ratio was 1:1.2. In addition, 6 individuals with asymptomatic malaria were detected in the community through an active case finding campaign. No cases were however reported on Bhutan side of the border. Anopheles culicifacies and An. annularis were identified from light trap collection. Saralpara is a remote and forested area where most of the residents are poor and engaged predominantly in farming activities. Being a harvesting season, they tend to stay overnight to protect their harvest from elephants. The health authorities in India and Bhutan exchanged information regularly at district-to-district level on the outbreak including through available and easy modes such as WhatsApp, telephone as well as electronic mail and collaborated in coherently implementing preventive measures on both sides and in managing the cases. The outbreak of malaria in Saralpara demonstrates the risk of malaria outbreak in remote and forested areas, with possibility of spill over to the other side of the border. The fact that two countries had excellent inter-country cross-border collaboration and prior commitment thereby preventing further transmission of malaria to Bhutan can serve as a best practice for cross-border control of malaria and other communicable diseases.


[Just as important as blood sugar control by drugs. A healthy cuisine for diabetics].
A modern optimal diet for diabetics offers more than simple control of blood sugar. Rather, it also aims to minimize the additional cardiovascular risk factors that are often present: hypertension and lipometabolic disorders as well as hyperuricemia. An improvement in the prognosis in particular of patients with a full-blown metabolic syndrome can be achieved only by comprehensively treating the various disorders associated with the disease. A specific "diabetic diet" is unknown. For the most part, recommendations focus on an energy-reduced, moderate-fat, high bulk, mixed diet. An additional aim is reduction of overweight with a BMI target of < 25 kg/m2. Since the eating habits of German diabetics are currently far from complying with these recommendations, greater engagement is required not only from the patient, but also from the care-providing physician. The latter should ensure that the patient receives qualified instruction, which can appreciably improve the patient's motivation and cooperation.


Targeting NRF2, Regulator of Antioxidant System, to Sensitize Glioblastoma Neurosphere Cells to Radiation-Induced Oxidative Stress.
The presence of glioma stem cells (GSCs), which are enriched in neurospheres, may be connected to the radioresistance of glioblastoma (GBM) due to their enhanced antioxidant defense and elevated DNA repair capacity. The aim was to evaluate the responses to different radiation qualities and to reduce radioresistance of U87MG cells, a GBM cell line. U87MG cells were cultured in a 3D model and irradiated with low (24 mGy/h) and high (0.39 Gy/min) dose rates of low LET gamma and high LET carbon ions (1-2 Gy/min). Thereafter, expression of proteins related to oxidative stress response, extracellular 8-oxo-dG, and neurospheres were determined. LD50 for carbon ions was significantly lower compared to LD50 of high and low dose rate gamma radiation. A significantly higher level of 8-oxo-dG was detected in the media of cells exposed to a low dose rate as compared to a high dose rate of gamma or carbon ions. A downregulation of oxidative stress proteins was also observed (NRF2, hMTH1, and SOD1). The NRF2 gene was knocked down by CRISPR/Cas9 in neurosphere cells, resulting in less self-renewal, more differentiated cells, and less proliferation capacity after irradiation with low and high dose rate gamma rays. Overall, U87MG glioma neurospheres presented differential responses to distinct radiation qualities and NRF2 plays an important role in cellular sensitivity to radiation.


Towards immunotherapeutic drugs and vaccines against multiple sclerosis.
Multiple sclerosis (MS) is an autoimmune, demyelinating disease of the central nervous system. Numerous treatment options are available to MS patients; however, these options need to be improved. Herein, we review the current drugs and therapeutic approaches available to MS patients, preclinical trial interventions and recent animal model studies for the potential therapy of MS. Since the current treatment of MS remains elusive and is limited, animal studies and clinical research offers an optimistic outlook.


Illness-related behaviour and utilization of oral health services among adult city-dwellers in Burkina Faso: evidence from a household survey.
In sub-Saharan Africa, the availability and accessibility of oral health services are seriously constrained and the provision of essential oral care is limited. Reports from the region show a very low utilization of oral health care services, and visits to dental-care facilities are mostly undertaken for symptomatic reasons. The objectives of the present study were to describe the prevalence of oral symptoms among adults in Ouagadougou, capital city of Burkina Faso and the use of oral health services and self-medication in response to these symptoms and to measure the associations between predisposing, enabling and needs factors and decisions to seek oral health care. The conceptual design of the study was derived from both the Andersen-Newman model of health care utilization and the conceptual framework of the WHO International Collaborative Study of Oral Health Outcomes. Data were obtained by two-stage stratified sampling through four areas representative of different stages of urbanization of Ouagadougou. The final study population comprised 3030 adults aged 15 years or over and the response rate was 65%. Overall, 28% of the respondents had experienced an oral health problem during the past 12 months; a high proportion (62%) reported pain or acute discomfort affecting daily life. In response to symptoms, only 28% used oral health facilities, 48% used self-medication and 24% sought no treatment at all. Multivariate analyses revealed that several socio-economic and socio-cultural factors such as religious affiliation, material living conditions and participation in a social network were significantly associated with the use of oral health care services by adults who had experienced oral health problems during the previous year. The proportion of people who have obtained oral health care is alarmingly low in Ouagadougou and self-medication appears to be an important alternative source of care for adult city-dwellers. Decision-makers in sub-Saharan countries must seek to ensure that access to essential oral health care is improved.


A descriptive analysis of midwifery education, regulation and association in 73 countries: the baseline for a post-2015 pathway.
Education, regulation and association (ERA) are the supporting pillars of an enabling environment for midwives to provide quality care. This study explores these three pillars in the 73 low- and middle-income countries who participated in the State of the World's Midwifery (SoWMy) 2014 report. It also examines the progress made since the previous report in 2011. A self-completion questionnaire collected quantitative and qualitative data on ERA characteristics and organisation in the 73 countries. The countries were grouped according to World Health Organization (WHO) regions. A descriptive analysis was conducted. In 82% of the participating countries, the minimum education level requirement to start midwifery training was grade 12 or above. The average length of training was higher for direct-entry programmes at 3.1 years than for post-nursing/healthcare provider programmes at 1.9 years. The median number of supervised births that must be conducted before graduation was 33 (range 0 to 240). Fewer than half of the countries had legislation recognising midwifery as an independent profession. This legislation was particularly lacking in the Western Pacific and South-East Asia regions. In most (90%) of the participating countries, governments were reported to have a regulatory role, but some reported challenges to the role being performed effectively. Professional associations were widely available to midwives in all regions although not all were exclusive to midwives. Compared with the 2011 SoWMy report, there is evidence of increasing effort in low- and middle-income countries to improve midwifery education, to strengthen the profession and to follow international ERA standards and guidelines. However, not all elements are being implemented equally; some variability persists between and within regions. The education pillar showed more systematic improvement in the type of programme and length of training. The reinforcement of regulation through the development of legislation for midwifery, a recognised definition and the strengthening of midwives' associations would benefit the development of other ERA elements and the profession generally.


Comparison of nuclear morphometric parameters in cytologic smears and histologic sections of spontaneous canine tumors.
Nuclear morphometry may provide useful diagnostic and prognostic information for neoplasms in animals. Most available data have been obtained from histologic sections. Nuclear morphometry of cytologic smears may provide important preoperative information. The goal of this study was to compare nuclear morphometric parameters in cytologic smears and histologic sections from spontaneous canine tumors. Mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear form factor (FF; nuclear perimeter(2)/4pi nuclear area) and their respective SDs were assessed by image analysis of both hematoxylin and eosin-stained histologic sections and May-Grünwald-Giemsa-stained cytologic smears from the same case in 20 spontaneous canine tumors of different histogenesis. The above parameters were selected as being the best morphometric tools for measuring variation in shape and size in cells after neoplastic transformation. Data were compared by ANOVA with P<.01 considered significant. There was a significant difference between histologic and cytologic specimens for MNA, MNP, and their SDs. Only the differences between FF and the SD of FF were not statistically significant. Only nuclear morphometric data related to nuclear shape and nuclear shape variability are comparable between histologic and cytologic specimens. Nuclear area and perimeter may be affected by the different fixation and smear preparation techniques used in histology and cytology.


Cytomegalovirus increases collagen synthesis in chronic rejection in the rat.
We have demonstrated previously that cytomegalovirus (CMV) infection enhances chronic renal allograft rejection in a rat model. Interstitial fibrosis, a characteristic finding for chronic rejection, was also more prominent in CMV-infected grafts. The effect of CMV on the development of fibrosis in this model was investigated here at the molecular level. The collagen/DNA ratio, gene expression of type I and III collagen mRNAs and the presence of myofibroblasts were examined. Transplantations were performed under triple drug immunosuppression in a rat strain combination of DA(RT1(a)) and BN(RT1(n)). One group of animals was infected with rat CMV and the other was left uninfected. The grafts were harvested at different time points post-transplantation. Graft histology was evaluated according to the Banff criteria and quantified by the chronic allograft damage index (CADI). Total collagen was measured and DNA and RNA were extracted from the grafts. Type I and III collagen mRNAs were determined by slot blot and in situ hybridizations. Myofibroblasts were demonstrated by immunohistochemistry. The time-related increase of the collagen/DNA ratio in the CMV-infected grafts was higher than in the uninfected animals, correlating with the development of fibrosis at the histology. The expression of type I and III collagen mRNAs peaked shortly after transplantation, together with the presence of myofibroblasts, with significantly higher peaks in the CMV-infected grafts compared with the non-infected ones. CMV increases the expression of both type I and III collagens and the accumulation of myofibroblasts, and enhances total collagen synthesis in the development of interstitial fibrosis in chronic renal allograft rejection.


Prevalence of glycemic variability and factors associated with the glycemic arrays among end-stage kidney disease patients on chronic hemodialysis.
Glycemic variability (GV) confers a significantly higher risk of diabetic-related complications, especially cardiovascular. Despite extensive research in this area, data on end-stage kidney disease (ESKD) patients on chronic hemodialysis are scarce. This study aims to determine the magnitude of GV among ESKD (diabetic vs nondiabetic) patients and its associated factors on hemodialysis days (HDD) and non-hemodialysis days (NHDD) where postulation of a higher GV observed among diabetic on HDD.We recruited 150 patients on hemodialysis, 93 patients with type 2 diabetic (DM-ESKD), and 57 with nondiabetic (NDM-ESKD). The GV indices (standard deviation [SD] and percentage coefficient variant [%CV]) were obtained from 11-point and 7-point self-monitoring blood glucose (fasting to post-meal) (SMBG) profiles on HDD and NHDD. The GV indices and its associated factors of both DM-ESKD and NDM-ESKD were analyzed to compare HDD vs NHDD.Mean blood glucose on HDD was 9.33 [SD 2.7, %CV 30.6%] mmol/L in DM-ESKD compared with 6.07 [SD 0.85, %CV 21.3%] mmol/L in NDM-ESKD (P = <.01). The DM-ESKD group experienced significantly above target GV indices compared to NDM-ESKD on both HDD and NHDD, particularly in the subgroup with HbA1c 8-10% (P = <.01). Presence of diabetes, older age, hyperlipidemia, HbA1c, ferritin levels, and albumin were identified as factors associated with GV.DM-ESKD patients have above-target GV indices, especially on HDD, therefore increasing their risk of developing future complications. We identified high HbA1c, older age group, presence of hyperlipidemia, ferritin levels, and albumin as factors associated with GV indices that may be used as surrogate markers for GV. Since these groups of patients are vulnerable to CVD mortality, urgent attention is needed to rectify it.


Phase II study of dose-adjusted EPOCH as initial therapy for adults with high-risk acute lymphoblastic leukemia.
Treatments for adults with newly-diagnosed acute lymphoblastic leukemia (ALL) may be prohibitively toxic and/or resource-intense. To address this, we performed a phase II study of dose-adjusted etoposide, prednisone, vincristine, cyclophosphamide, and doxorubicin (DA-EPOCH). Imatinib or dasatinib was added for Ph + disease; rituximab was added when CD20+. Fifty-three patients were evaluable: 28 with Ph + disease, and 25 with Ph-. All patients had ≥1 high-risk clinical feature. Measurable residual disease-negativity by multiparameter flow cytometry within 4 cycles was achieved in 71% in patients with Ph + ALL and 64% in Ph - ALL. Median overall survival (OS) was 49 months, with a 2-year OS of 71%. Median relapse-free survival (RFS) in the 47 patients that attained morphologic remission was 24 months, with a 2-year RFS of 57%. Early mortality was 2%. In summary, DA-EPOCH yields deep and durable remissions in adults with ALL comparable to some resource-intense strategies but with a low rate of treatment-related death.


Serial GH Measurement After Intravenous Catheter Placement Alone Can Detect Levels Above Stimulation Test Thresholds in Children.
GH stimulation testing is limited by poor specificity and reproducibility in identifying GH deficiency. Intravenous line placement (IVP) in pediatrics may be a stimulus for GH secretion. The objective of the study was to determine whether the measurement of GH at baseline as well as 15 and 30 minutes after IVP detects additional patients with sufficient peak GH concentrations who are not identified by a subsequent insulin tolerance test (ITT). The ITT protocol was modified to include GH measurement at the time of IVP (t = 0) and 15 (t = 15) and 30 (t = 30) minutes later. Insulin was administered at t = 30, and an ITT was performed as per standard protocols. Children were grouped according to the indication for ITT: initial evaluation of GH deficiency (group 1); and GH deficiency at transition to adulthood (group 2). Ninety-seven patients were included (76 in group 1, 21 in group 2). Of these, 27 (28%) had a peak GH concentration of 7 ng/mL or greater (19 in group 1, eight in group 2) either after IVP or ITT. Thirteen subjects (11 in group 1, two in group 2) had GH concentrations of 7 ng/mL or greater after IVP, without exceeding this on a subsequent ITT. Among the 11 group 1 patients, three of these GH peaks of 7 ng/mL or greater occurred at t = 0, 5 at t = 15, and 5 at t = 30, including one patient who had a peak GH of 7 ng/mL or greater at all three time points. Some children will not have a sufficient GH response to pharmacological stimuli but will have a robust response to IVP. We recommend GH measurement after IVP in children undergoing GH stimulation testing, particularly when there is a delay between IVP and the administration of the pharmacological stimulus.


Weight loss interventions in living donor liver transplantation as a tool in expanding the donor pool: A systematic review and meta-analysis.
With increasing rates of liver transplantation and a stagnant donor pool, the annual wait list removals have remained high. Living donor liver transplantation (LDLT) is an established modality in expanding the donor pool and is the primary method of liver donation in large parts of the world. Marginal living donors, including those with hepatic steatosis, have been used to expand the donor pool. However, due to negative effects of steatosis on graft and recipient outcomes, current practice excludes overweight or obese donors with more than 10% macro vesicular steatosis. This has limited a potentially important source to help expand the donor pool. Weight loss is known to improve or resolve steatosis and rapid weight loss with short-term interventions have been used to convert marginal donors to low-risk donors in a small series of studies. There is, however, a lack of a consensus driven standardized approach to such interventions. To assess the available data on using weight loss interventions in potential living liver donors with steatotic livers and investigated the feasibility, efficacy, and safety of using such donors on the donor, graft and recipient outcomes. The principal objective was to assess if using such treated donor livers, could help expand the donor pool. We performed a comprehensive literature review and meta-analysis on studies examining the role of short-term weight loss interventions in potential living liver donors with hepatic steatosis with the aim of increasing liver donation rates and improving donor, graft, and recipient outcomes. A total of 6 studies with 102 potential donors were included. Most subjects were males (71). All studies showed a significant reduction in body mass index post-intervention with a mean difference of -2.08 (-3.06, 1.10, I 2 = 78%). A significant reduction or resolution of hepatic steatosis was seen in 93 of the 102 (91.2%). Comparison of pre- and post-intervention liver biopsies showed a significant reduction in steatosis with a mean difference of -21.22 (-27.02, -15.43, I 2 = 56%). The liver donation rates post-intervention was 88.5 (74.5, 95.3, I 2 = 42%). All donors who did not undergo LDLT had either recipient reasons or had fibrosis/steatohepatitis on post intervention biopsies. Post-operative biliary complications in the intervention group were not significantly different compared to controls with an odds ratio of 0.96 [(0.14, 6.69), I 2 = 0]. The overall post-operative donor, graft, and recipient outcomes in treated donors were not significantly different compared to donors with no steatosis. Use of appropriate short term weight loss interventions in living liver donors is an effective tool in turning marginal donors to low-risk donors and therefore in expanding the donor pool. It is feasible and safe, with comparable donor, graft, and recipient outcomes, to non-obese donors. Larger future prospective studies are needed.


Sustained Release of Phosphates From Hydrogel Nanoparticles Suppresses Bacterial Collagenase and Biofilm Formation in vitro.
Intestinal disease or surgical intervention results in local changes in tissue and host-derived factors triggering bacterial virulence. A key phenotype involved in impaired tissue healing is increased bacterial collagenase expression which degrades intestinal collagen. Antibiotic administration is ineffective in addressing this issue as it inadvertently eliminates normal flora while allowing pathogenic bacteria to "bloom" and acquire antibiotic resistance. Compounds that could attenuate collagenase production while allowing commensal bacteria to proliferate normally would offer major advantages without the risk of the emergence of resistance. We have previously shown that intestinal phosphate depletion in the surgically stressed host is a major cue that triggers P. aeruginosa virulence which is suppressed under phosphate abundant conditions. Recent findings indicate that orally administered polyphosphate, hexametaphosphate, (PPi) suppresses collagenase, and biofilm production of P. aeruginosa and S. marcescens in animal models of intestinal injury but does not attenuate E. faecalis induced collagenolytic activity (Hyoju et al., 2017). Systemic administration of phosphates, however, is susceptible to rapid clearance. Given the diversity of collagenase producing bacteria and the variation of phosphate metabolism among microbial species, a combination therapy involving different phosphate compounds may be required to attenuate pathogenic phenotypes. To address these barriers, we present a drug delivery approach for sustained release of phosphates from poly(ethylene) glycol (PEG) hydrogel nanoparticles. The efficacy of monophosphate (Pi)- and PPi-loaded NPs (NP-Pi and NP-PPi, respectively) and a combination treatment (NP-Pi + NP-PPi) in mitigating collagenase and biofilm production of gram-positive and gram-negative pathogens expressing high collagenolytic activity was investigated. NP-PPi was found to significantly decrease collagenase and biofilm production of S. marcescens and P. aeruginosa. Treatment with either NP-Pi or NP-Pi + NP-PPi resulted in more prominent decreases in E. faecalis collagenase compared to NP-PPi alone. The combination treatment was also found to significantly reduce P. aeruginosa collagenase production. Finally, significant attenuation in biofilm dispersal was observed with NP-PPi or NP-Pi + NP-PPi treatment across all test pathogens. These findings suggest that sustained release of different forms of phosphate confers protection against gram-positive and gram-negative pathogens, thereby providing a promising treatment to attenuate expression of tissue-disruptive bacterial phenotypes without eradicating protective flora over the course of intestinal healing.


[Role of HMGB1-RAGE/TLRs-NF-κB signaling pathway on bone mesenchymal stem cells transplantation therapy for lipopolysaccaride-induced coagulation disorder rats].
To determine the effect of bone mesenchymal stem cells (BMSCs) in transplantation therapy for lipopolysaccharide (LPS)-induced coagulation disorder and the underlying mechanism of high mobility group protein B1-receptors for advanced glycation end products/Toll-like receptors-nuclear factor-κB (HMGB1-RAGE/TLRs-NF-κB) signaling pathway. BMSCs of female Sprague-Dawley (SD) rats ageing 4-5 weeks old were extracted and cultivated in vitro, and the fourth-passaged BMSCs phenotype was identified by flow cytometry for transplantation in the following experimental study. The rats were randomly divided into normal saline (NS) control group, LPS group, and BMSC group according to the random number table with 15 rats in each group. Coagulation disorders model was reproduced by injection of 1 mg/kg LPS via saphenous vein, and the rats in the NS control group was injected with equal volume NS. Those in the BMSC group were infused BMSC 0.5 mL containing 1×106 cells via tail vein at 2 hours after LPS injection, and the rats in other groups were injected with equal volume NS. Abdominal aorta blood was collected at 1, 3 and 7 days post operation. Coagulation indexes such as platelet count (PLT), platelet volume distribution width (PDW), mean platelet volume (MPV), plateletcrit (PCT), platelet large cell ratio (P-LCR), activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), international normalized ratio (INR), and fibrinogen (FIB) were determined. The mRNA levels and contents of HMGB1, RAGE, TLR2/4 and NF-κB were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. (1) The cells cultured in vitro were spindle shaped or flat. The fourth-passaged BMSCs phenotype was successfully identified by flow cytometry technology. (2) Coagulation indexes: compared with NS control group, PLT, PCT and FIB in LPS group were significantly decreased, PDW, MPV, P-LCP, and INR were significantly increased, and APTT, PT, and TT were significantly prolonged from the first day. Furthermore, those in LPS group were gradually ameliorated with prolongation of LPS induction time. The coagulation function abnormality induced by LPS was reversed by BMSCs with significant difference at 1 day as compared with LPS group [PLT (×109/L): 398.8±17.9 vs. 239.1±15.8, PCT (%): 0.35±0.04 vs. 0.23±0.06, FIB (g/L): 1.7±0.6 vs. 0.8±0.1, PDW (%): 12.4±1.6 vs. 16.2±1.5, MPV (fl): 11.0±1.6 vs. 13.7±1.1, P-LCP (%): 13.0±2.1 vs. 15.3±2.7, INR: 1.52±0.17 vs. 1.82±0.19, APTT (s): 66.3±4.1 vs. 89.5±4.5, PT (s): 18.3±0.7 vs. 25.1±1.9, TT (s): 87.5±7.8 vs. 115.0±9.7, all P < 0.05], till 7 days. (3) HMGB1-RAGE/TLRs-NF-κB signaling pathway related molecules: compared with NS control group, the mRNA expressions and contents of HMGB1, RAGE, TLR2/4 and NF-κB were significantly increased in LPS group from the first day. However, the mRNA expressions and contents of the molecules in LPS group were gradually decreased with prolongation of LPS induction time. After BMSC intervention, the mRNA expressions and contents of molecules at 1 day were significantly lower than those of LPS group [HMGB1 mRNA (2-ΔΔCt): 10.77±0.04 vs. 24.51±3.69, HMGB1 content (μg/L): 0.48±0.01 vs. 0.95±0.06; RAGE mRNA (2-ΔΔCt): 11.57±1.11 vs. 18.08±0.29, RAGE content (μg/L): 0.73±0.04 vs. 1.37±0.06; TLR2 mRNA (2-ΔΔCt): 2.60±0.22 vs. 12.61±0.27, TLR2 content (μg/L): 0.81±0.03 vs. 1.59±0.09; TLR4 mRNA (2-ΔΔCt): 2.95±0.52 vs. 4.06±0.11, TLR4 content (μg/L): 0.80±0.09 vs. 1.18±0.11; NF-κB mRNA (2-ΔΔCt): 1.29±0.06 vs. 7.79±0.25, NF-κB content (μg/L): 1.22±0.24 vs. 2.42±0.26, all P < 0.05], till 7 days. BMSCs administration could ameliorate the coagulation function in LPS-induced coagulation disorder rats and these might be associated with HMGB1-RAGE/TLRs-NF-κB signaling pathway inhibition.


The history of interventional ultrasound.
Ultrasound has been used to guide interventional procedures for more than 30 years. Initial applications included biopsy techniques and simple aspiration of fluid collections. However, with improved sonographic imaging and the use of different ultrasonic probes as well as development of less invasive therapies, there has been an increase in the use of ultrasound to guide interventional procedures. Interventional ultrasound has become routine in most medical specialties. Because of the remarkable success of interventional procedures guided by sonography, combined with an outstanding safety record, there is no doubt that there will be a future increase in the number and types of interventional procedures performed under sonographic guidance.


Combined Fluorescence-Based in Vitro Assay for the Simultaneous Detection of Cell Viability and Alkaline Phosphatase Activity during Osteogenic Differentiation of Osteoblast Precursor Cells.
Novel bone substitute materials need to be evaluated in terms of their osteogenic differentiation capacity and possible unwanted cytotoxic effects in order to identify promising candidates for the therapy of bone defects. The activity of alkaline phosphatase (ALP) is frequently quantified as an osteogenic marker, while various colorimetric assays, like MTT assay, are used to monitor cell viability. In addition, the DNA or protein content of the samples needs to be quantified for normalization purposes. As this approach is time consuming and often requires the analysis of multiple samples, we aimed to simplify this process and established a protocol for the combined fluorescence-based quantification of ALP activity and cell viability within one single measurement. We demonstrate that the fluorogenic substrate 4-methylumbelliferone-phosphate (4-MUP) and the commonly used para-nitrophenylphosphate (p-NPP) produce comparable and highly correlating results. We further show that fluorescein-diacetate (FDA) can be used to quantify both cell viability and cell number without interfering with the quantification of ALP activity. The measurement of additional normalization parameters is, therefore, unnecessary. Therefore, the presented assay allows for a time-efficient, simple and reliable analysis of both ALP activity and cell viability from one sample and might facilitate experiments evaluating the osteogenic differentiation of osteoblast precursor cells.


Temperature mapping of laser-induced hyperthermia in an ocular phantom using magnetic resonance thermography.
Laser-induced heating in an ocular phantom is measured with magnetic resonance thermography (MRT) using temperature-dependent phase changes in proton resonance frequency. The ocular phantom contains a layer of melanosomes isolated from bovine retinal pigment epithelium. The phantom is heated by the 806-nm output of a continuous wave diode laser with an irradiance of 2.4 to 21.6 W/cm2 in a beam radius of 0.8 or 2.4 mm, depending on the experiment. MRT is performed with a 2 T magnet, and a two-turn, 6-cm-diam, circular radio frequency coil. Two-dimensional temperature gradients are measured within the plane of the melanin layer, as well as normal to it, with a temperature resolution of 1 degrees C or better. The temperature gradients extending within the melanin layer are broader than those orthogonal to the layer, consistent with the higher optical absorption and consequent heating in the melanin. The temperature gradients in the phantom measured by MRT closely approximate the predictions of a classical heat diffusion model. Three-dimensional temperature maps with a spatial resolution of 0.25 mm in all directions are also made. Although the temporal resolution is limited in the prototype system (22.9 s for a single image "slice"), improvements in future implementations are likely. These results indicate that MRT has sufficient spatial and temperature resolution to monitor target tissue temperature during transpupillary thermotherapy in the human eye.


HSV-induced reactivation: contribution of epinephrine after corneal iontophoresis.
A modified, bilateral iontophoresis technique was used to evaluate herpes simplex virus (HSV) reactivation and epinephrine distribution and concentration in full-thickness corneas and in trigeminal ganglia of rabbits. Infectious virus was recovered from 58% of epinephrine-induced eyes 24-96 hours after iontophoresis. A histochemical adrenochrome oxidation method localized epinephrine exclusively in the corneal epithelium and anterior stromal lamellae after iontophoresis. Epinephrine introduced iontophoretically into the corneal epithelium and stroma did not reach the ganglion level as detected by the assay. Radiometric-enzymatic assay of corneal and ganglionic tissues demonstrated a transient increase in total corneal catecholamine levels from an average of 417 micrograms to an average of 778 micrograms ; concentrations returned to approximate control values 24 hours after iontophoresis. Ganglion total catecholamine levels increased from an average of 606 micrograms to an average of 1211 micrograms for a 12-hour period and remained elevated at 24 hours after iontophoresis. Epinephrine acting directly on the corneal epithelium and epinephrine metabolites at the ganglion level could act synergistically to induce ocular and ganglionic HSV reactivation.


Diagnostic, therapeutic and health-care management protocol for major abdominal trauma at the "Santa Maria" Hospital of Terni. Analysis of the results after two years.
Trauma is the fourth leading cause of death and is more common in people under 45 age. Abdominal trauma is cause of death in 7-10% of traumatized patients. Aim is to evaluate the effectiveness of the diagnostic, therapeutic and health-care management protocol for major abdominal trauma. A prospective study called DGR Azienda Ospedaliera S. Maria Terni n. 159. was registered on February 2014. Patients with abdominal major trauma admitted at Santa Maria Hospital of Terni from January 2015 to December 2016 were considered in this paper. Patients' demographics, comorbidities, application of the operative and non operative management (OM-NOM), Mortality, effectiveness of diagnostics were investigated. The most of the patients were the elderly. Staff members were compliant and operated in conformity to the protocol for abdominal trauma. Non operative management (NOM) was performed in 63% of patients, according to data from the literature. On the other hand data with respect to the performing of diagnostic examinations as Focused assessment with ultrasonography for trauma (FAST), chest X-ray, Computed Tomography (CT) were discordant with literature. No patient underwent damage control surgery (DCS). Number of missed injuries (3.3%) and overall (13.3%), daily and weekend mortality were similar to those from the available literature. In our experience, the best results were found regarding the sensitivity of the FAST and CT in unstable and stable patients respectively, and the lower incidence of: NOM failure (NOM 0%, NOM plus endovascular treatment 14.4%) , mortality in stable patients (2.3%) and night mortality (5%). Despite the application of trauma protocol at Terni Hospital has been effective it can be further improved. Diagnostic, Emergency surgery, Major Abdominal Trauma, Therapeutic and health-care management protocol.


The Monocyte to High-Density Lipoprotein Cholesterol Ratio Is Associated with Left Ventricular Diastolic Function in Patients with No Significant Perfusion Abnormality.
The monocyte to high-density lipoprotein cholesterol (HDL-C) ratio has been considered to be a prognostic marker. Whether this ratio is associated with left ventricular (LV) diastolic function remains undetermined. We tested the hypothesis that the monocyte to HDL-C ratio is associated with LV diastolic parameters derived from gated myocardial perfusion single-photon emission computed tomography (SPECT) in patients with no significant perfusion abnormality.The study population included 196 patients with no significant perfusion abnormalities and preserved ejection fraction. The peak filling rate (PFR) and one-third mean filling rate (1/3 MFR) were obtained as LV diastolic parameters using gated SPECT. Monocyte counts and plasma HDL-C levels were also examined.Significant associations were observed between the monocyte to HDL-C ratio and PFR (r = -0.20; P = 0.005) and 1/3 MFR (r = -0.19; P = 0.009). Multivariate linear regression analysis was performed to determine factors associated with LV diastolic parameters. Age (β = -0.27; P < 0.001), LV end-diastolic volume (β = -0.19; P = 0.034), and monocyte to HDL-C ratio (β = -0.15; P = 0.027) were determined to be significantly associated with PFR. Moreover, age (β = -0.13; P = 0.007), LV mass index (β = -0.18; P = 0.037), and the monocyte to HDL-C ratio (β = -0.13; P = 0.045) were significantly associated with 1/3 MFR.These results demonstrated that the monocyte to HDL-C ratio is associated with LV diastolic function, as derived from gated SPECT in patients with no significant perfusion abnormality.


The use of computer generated patient profiles to evaluate resident performance in patient care.
This paper describes the way in which data from a computer-based health information system are used to review the service experiences of family practice residents. First, it discusses the development of the patient profiles that provide a chronological account of a patient's visits, their purposes, diagnoses, laboratory procedures, treatments, and outcomes. Then, through four cases, it describes the way in which these computer-generated displays are used by faculty to conduct concurrent reviews of residents' performances, to select medical records for review, and to initiate feedback and instruction to residents as they care for their patients.


Parochial cooperation in nested intergroup dilemmas is reduced when it harms out-groups.
In intergroup settings, humans often contribute to their in-group at a personal cost. Such parochial cooperation benefits the in-group and creates and fuels intergroup conflict when it simultaneously hurts out-groups. Here, we introduce a new game paradigm in which individuals can display universal cooperation (which benefits both in- and out-group) as well as parochial cooperation that does, versus does not hurt the out-group. Using this set-up, we test hypotheses derived from group selection theory, social identity, and bounded generalized reciprocity theory. Across three experiments we find, first, that individuals choose parochial over universal cooperation. Second, there was no evidence for a motive to maximize differences between in- and out-group, which is central to both group selection and social identity theory. However, fitting bounded generalized reciprocity theory, we find that individuals with a prosocial value orientation display parochial cooperation, provided that this does not harm the out-group; individualists, in contrast, display parochialism whether or not nut it hurts the out-group. Our findings were insensitive to cognitive taxation (Experiments 2-3), and emerged even when universal cooperation served social welfare more than parochialism (Experiment 3). (PsycINFO Database Record


Executive rounding program improves communication, performance.
A patient safety executive rounding program at St. Charles Hospital, Port Jefferson, NY, has united staff and helped to identify a number of performance issues. The success of the program has even surprised top administrators who pushed to have it implemented. Each month, teams of executive staff and department heads make rounds through all areas of the hospital, talking with staff, identifying problem areas and noting best practices that could be used in other departments.


A rapid, novel model of culturing cranial nerve X-derived motoneurons for screening trophic factor outgrowth response.
After cranial nerve X (CN X) injury, vocal fold paralysis treatments currently face a myriad of obstacles in achieving non-synkinetic, functional reinnervation. Of particular therapeutic interest is the targeted administration of locally expressed biological neurotrophic factors (NFs). To date, a method to culture mature CN X motoneurons for NF responsiveness screening has not been described. We herein present a novel method for establishing mature murine CN X motoneuron cultures, and use the model to test CN X motoneuron outgrowth response to individual and paired ascending concentrations of selected neurotrophic factors [glial cell-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and ciliary neurotrophic factor (CNTF)]. Findings demonstrated low concentration (5 ng/ml) CNTF to have the greatest positive effect on motoneuron outgrowth, beyond that of both indivual NF and paired NF combinations, based on total neurite outgrowth [mean total neurite outgrowth = 445.7±84.45 μm in the (5 ng/ml) CNTF group versus 179.7±13.63 μm in saline controls (P<0.01)]. Paired treatments with CNTF/GDNF, and CNTF/BDNF promoted motoneuron branching at a variety of concentrations beyond saline controls, and paired GDNF/BDNF had inhibitory effects on motoneuron branching. Our described in vitro model of establishing mature CN X cultures allowed rapid screening for responsiveness to therapeutic NFs at a variety of concentrations and combinations. While the model ultimately may be used to investigate the molecular mechanisms of CN X motoneuron regeneration, the current study identified CNTF as a promising therapeutic candidate for the promotion of CN X outgrowth.


Cholinesterase activity in serum during general anesthesia in patients with or without vascular disease.
Maintaining hemodynamic stability during the induction and maintenance of anesthesia is one of the challenges of the anesthesiologist. Patients with vascular disease are at increased risk of instability due to imbalance between the sympathetic and parasympathetic parts of the autonomic nervous system, a balance accessible by serum cholinesterase activity. We aim to characterize the dynamics of cholinesterase activity in patients undergoing general anesthesia (GA) and surgery. This was a prospective study of 57 patients undergoing ambulatory or vascular surgery under GA. Cholinesterase activity was measured before the induction of anesthesia, after 15 min and at the end of surgery by calculating the capacity of serum acetylcholinesterase (AChE) and butyrylcholinesterase to hydrolyze AcetylThioCholine. Data on atherosclerotic disease, anesthesia management were analyzed. Both AChE and total cholinergic status (CS) decreased significantly after GA induction at 15 min and even more so by the end of surgery. Vascular surgery patients had lower baseline cholinesterase activity compared to ambulatory surgery patients. Patients requiring intraoperative administration of phenylephrine for hemodynamic support (21.1%) had a significantly lower level of AChE and CS compared to untreated patients. Our findings serve as a mirror to the sympathetic/parasympathetic imbalance during GA, with a marked decrease in the parasympathetic tone. The data of a subgroup analysis show a correlation between low cholinesterase activity and an increase in the need for hemodynamic support.


[Experimental systemic enzyme therapy of gouty and primary glomerulonephritis].
The influence of a systemic enzymotherapy on the morphological, biochemical, and functional manifestations of the kidney damage during the experimental gouty and primary glomerulonephritis is described in comparison to the results obtained by traditional methods.


Freedom-to-operate analysis of a transgenic multivitamin corn variety.
In this article, we explore the intellectual property (IP) landscape relevant to the production and commercialization of Carolight(™) , a transgenic multivitamin corn variety created on humanitarian grounds to address micronutrient deficiencies in low-and-middle-income countries. The successful production of this variety requires IP rights risk management because there is a strong protection on inventions and processes via patent portfolios in both developing and industrialized countries. The IP framework is complex, and specialist patent lawyers are usually employed to perform such analysis, but the costs cannot always be met by small, publicly funded projects. We report an alternative strategy, a do-it-yourself patent analysis, to produce a review with limited legal value that can nevertheless lay the foundations for a subsequent more in-depth professional freedom-to-operate opinion.


[Study on the influence of maternal physiology, mentality and prenatal health care on children born with low birth weight].
To explore the role of maternal physiology, mentality and prenatal health care on low birth weight (LBW). A LBW-small for gestation age (SGA)-control group retrospective study was conducted based upon a population of infants born from November 2004 to June 2006. 896 cases involved in this study were entirely under voluntary participation and subject could withdraw their consent at any point. All subject recruitment and enrollment took place at Fujian Provincial Maternal and Children Hospital at the time of delivery. Using chi2 test for single factor analysis and logistic regression for multiple analyses. There were 15 out of 22 single factors having statistical significances, including 2 maternal psychological, 4 physiological, 6 pathological and 3 prenatal health care factors respectively. When multi-factorial stepwise regression analyses was performed, there were 9 factors for main relative factors of LBW, including women's height, number of prenatal examination, in-normal non-stress test, umbilical cord around the neck, retardation of the umbilical blood flow, week of gestation when the first examination was performed, premature rupture of membrane, preference on the sex of the infant, abnormal family history etc. were influencing the existence of LBW. Awareness on health information appeared to be a protective factor, suggesting that LBW could be prevented during the pregnant period. Multiple factors seemed to be contributing to the incidence of low birth weight.


Homocysteine metabolism, growth performance, and immune responses in suckling and weanling piglets.
Homocysteine (Hcy), an intermediary sulfur AA, is recognized as a powerful prooxidant with deleterious effects on physiological and immune functions. In piglets, there is an acute 10-fold increase of plasma concentrations of homocysteine (pHcy) during the first 2 wk of life. This project aimed to maximize pHcy variations within physiological ranges using typical supplies of folates and vitamin B12 (B12) to sows and piglets. Growth, immune response, and Hcy metabolism of piglets were studied until piglets reached 56 d of age. Third-parity sows were randomly assigned to a 2 × 2 split-plot design with 2 dietary treatments during gestation and lactation, S(-) (1 mg/kg folates and 20 µg/kg B12, n = 15) and S(+) (10-fold S(-) levels, n = 16), and 2 treatments to piglets within each half litter, intramuscular injections (150 µg) of B12 (P(+)) at d 1 and 21 (weaning) and saline (P(-)). Within each litter of 12 piglets, 3 P(+) and 3 P(-) piglets were studied for growth and Hcy metabolism, and the others were studied for immune responses. During lactation, plasma B12 decreased and was transiently greater in S(+) vs. S(-) piglets on d 1 and P(+) vs. P(-) piglets on d 7 (sow treatment × age and piglet treatment × age; P < 0.05). From 14 to 21 d of age, pHcy was 33% lower in S(+)P(+) vs. S(-)P(-) piglets (sow treatment × piglet treatment interaction; P < 0.05). At 56 d of age, hepatic B12 was greater and pHcy was lower for P(+) vs. P(-) piglets (P < 0.05). No treatment effect was observed on growth except for a lower postweaning G:F in S(+)P(-) piglets than in others (sow treatment × piglet treatment interaction; P < 0.05). Positive correlations were observed between pHcy and growth (r > 0.29, P < 0.05) before and after weaning. Antibody responses to ovalbumin and serum tumor necrosis factor-α were not affected by treatments, but postweaning serum IL-8 peaked earlier in S(-)P(-) vs. S(+)P(+) piglets (piglet treatment × age; sow treatment × piglet treatment interaction, P < 0.05). Proliferation of lymphocytes in response to the mitogen concanavalin A tended to be lower in culture media supplemented with sera from S(-) vs. S(+) piglets (P = 0.081) and P(-) vs. P(+) piglets (P = 0.098), and the reduction of response was more marked (P < 0.05) with high (>21 µM) compared to medium (17 to 21 µM) or low (<17 µM) pHcy. In conclusion, the present vitamin supplements to sows and/or piglets produced variations of pHcy that were not apparently harmful for growth performance of piglets. The greater pHcy, particularly prevalent in S(-) and/or P(-) piglets, had negative effects on some indicators of immune responses, suggesting that these young animals may be immunologically more fragile.


Airborne Precursors Predict Maternal Serum Perfluoroalkyl Acid Concentrations.
Human exposure to persistent perfluoroalkyl acids (PFAAs), including perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), and perfluorooctanesulfonate (PFOS), can occur directly from contaminated food, water, air, and dust. However, precursors to PFAAs (PreFAAs), such as dipolyfluoroalkyl phosphates (diPAPs), fluorotelomer alcohols (FTOHs), perfluorooctyl sulfonamides (FOSAs), and sulfonamidoethanols (FOSEs), which can be biotransformed to PFAAs, may also be a source of exposure. PFAAs were analyzed in 50 maternal sera samples collected in 2007-2008 from participants in Vancouver, Canada, while PFAAs and PreFAAs were measured in matching samples of residential bedroom air collected by passive sampler and in sieved vacuum dust (<150 μm). Concentrations of PreFAAs were higher than for PFAAs in air and dust. Positive associations were discovered between airborne 10:2 FTOH and serum PFOA and PFNA and between airborne MeFOSE and serum PFOS. On average, serum PFOS concentrations were 2.3 ng/mL (95%CI: 0.40, 4.3) higher in participants with airborne MeFOSE concentrations in the highest tertile relative to the lowest tertile. Among all PFAAs, only PFNA in air and vacuum dust predicted serum PFNA. Results suggest that airborne PFAA precursors were a source of PFOA, PFNA, and PFOS exposure in this population.


Effect of endotoxin on oleic acid lung injury does not depend on priming.
Recent studies have demonstrated significant synergistic physiological and biochemical effects between low-dose endotoxin (Etx) administration and oleic acid (OA)-induced canine lung injury. To evaluate whether this interaction depends on Etx priming of some key cell population, we compared the effects of giving low-dose Etx both after as well as before inducing lung injury with OA. In addition to hemodynamic and blood-gas measurements, positron emission tomographic imaging was used to measure edema accumulation and intrapulmonary blood flow distribution. Biochemical measurements of the stable metabolites of prostacyclin and thromboxane were obtained as well as measurements of isoprostanes and reactive sulfhydryls as evidence for possible concomitant oxidant production. We found that the physiological and biochemical effects of low-dose Etx developed 30-45 min after its administration, regardless of whether Etx was administered before or after OA. No increase in either isoprostane or reactive sulfhydryl production after Etx and/or OA was detected. These data suggest that the synergistic effect of low-dose Etx and OA-induced lung injury is not due to a priming effect of Etx.


Experience of recombinant activated factor VII usage during surgery in patients with haemophilia with inhibitors.
Inhibitor development is one of the most challenging complications of haemophilia management. Haemostatic control in patients with haemophilia with inhibitors can be difficult, and is especially risky in those undergoing surgical interventions. Most haemophilia patients with inhibitors suffer from chronic joint disease requiring surgical correction due to recurrent bleeding episodes. The aim of this study was to assess the use of recombinant activated factor VII (rFVIIa) as haemostatic therapy during orthopaedic surgery in haemophilia patients with inhibitors. A series of case reports was retrospectively collected to describe clinical experience of rFVIIa use in inhibitor patients undergoing a range of orthopaedic surgical procedures at a single centre. All surgeries were performed using standard methods. All patients received rFVIIa at a starting dose of 120 μg kg(-1) with the subsequent regimens depending on the type of surgery. rFVIIa provided effective haemostasis in 23 patients with haemophilia A and inhibitors (15 with high inhibitor titres) undergoing orthopaedic surgery. The majority (70%) of surgical procedures were major (joint and extra-articular surgery). The doses and intervals of rFVIIa treatment used varied depending on the severity of bleeding, and the type (major or minor) or site of surgery. In all cases, administration of rFVIIa achieved good haemostasis. In all 23 patients with haemophilia with inhibitors, rFVIIa treatment in orthopaedic interventions proved to be an efficient haemostatic agent, providing effective intra-operative and postoperative haemostasis.


Throwing in the deep end: athletes, coaches and support staff experiences, perceptions and beliefs of upper limb injuries and training load in elite women's water polo.
To understand elite athlete, coach and support staff experiences, perceptions and beliefs in women's water polo with managing upper limb injuries and monitoring training loads. Inductive qualitative design. Twenty athletes, coaches and support staff were purposively recruited and participated in semistructured interviews. Participants either had experienced an upper limb injury or had experience managing athletes with upper limb injuries. Interviews were conducted in-person or virtually, audio-recorded, deidentified, transcribed verbatim and cleaned to ensure accuracy. Data were thematically analysed. Analysis identified five cohesive themes: (1) upper limb injury management is adequate-but prevention, communication and knowledge need improving, (2) current training load monitoring generates uncertainty and lack of consistency of processes-due to reliance on internal, and lack of external load monitoring, (3) optimal training load monitoring requires objective measurement of training load-that accurately measures the external load of athletes' upper limbs, (4) athlete-centred philosophy matters-including athlete-centred care to facilitate individually tailored rehabilitation programmes and their inclusion in management decisions, (5) mental, social and emotional aspects of upper limb injury management matter-acknowledging feelings of loss of team inclusion, fear of missing out and frustration felt by athletes as well as the emotional labour felt by coaches when supporting athletes with an upper limb injury. Upper limb injury management and training load monitoring are evolving areas where objective measurement of training load may assist in increasing consistency of communication, collaboration and coordination between all stakeholders, and to address uncertainty. Stakeholders placed value in intangible qualities such as trust and care in their relationships with other collaborators-facilitating athlete physical, mental and emotional recovery following upper limb injuries.


Nonlinear responses of degenerate two-level systems to intense few-cycle pulses.
Population transfers in degenerate (or almost degenerate) two-level systems interacting with the few-cycle laser pulse are investigated. A simple and analytical formula of nonadiabatic transition probability is derived with completely degenerate condition, demonstrating the sensitive dependence of the transition probability on the phase of the few-cycle pulses. As one of the applications of this formula, a new way of controlling the nuclear wave packet dynamics at a potential curve crossing by 1 cycle laser pulse is proposed.


[On organization of palliative medical care].
The study provides an analysis of the results of the questionnaire survey of physicians in the Russian Federation with the purpose of determining the competences of medical specialists and practice of palliative care and emerging problems. This article presents and discusses the data obtained during the study.


Comparative bacteriology of chronic blepharitis.
One hundred and fifteen patients with chronic blepharitis were compared with 47 normal controls. Six clinically distinct groups of blepharitis were observed: staphylococcal; seborrhoeic, alone, with associated staphylococcal superinfection, meibomian seborrhoea, or secondary inflammation of the meibomian glands; and meibomian keratoconjunctivitis (MKC). Staphylococcus aureus was isolated in appreciable frequency from the staphylococcal and the mixed staphylococcal/seborrhoeic groups in contrast to the normal and non-staphylococcal groups. Coagulase-negative Staphylococcus spp., Propionibacterium acnes, and cornyneform bacteria were the most commonly isolated bacteria from the lid for all groups. Cultures of material expressed from the meibomian glands yielded similar organisms but in reduced frequency. Testing of antibiotic susceptibility revealed Staph aureus to be usually sensitive to most commonly used ophthalmic antimicrobials except sulphonamides.


Proteolytic fragmentation of Helix pomatia alpha-hemocyanin: structural domains in the polypeptide chain.
alpha-Hemocyanin from the Roman snail Helix pomatia is composed of polypeptide chains with a molecular weight of 360000 +/- 30000. The cylindrically shaped hemocyanin molecule contains 20 of these large chains. The polypeptide chain has been split into components with molecular weights of: 210000, 154000, 147000, 112000, 120000, 98000, 55000, and 50000, by gentle proteolysis with enzymes of different specificities. Most of the fragments have molecular weights which are about 50000 or a multiple of 50000. Departure from these values, as found in the 112000 and 120000 fragments, is probably caused by the high carbohydrates content of these components. A mixture of these fragments has the same oxygen binding properties as the nondigested protein. Subtilisin converts the hemocyanin polypeptide chain, under appropriate conditions, almost completely into fragments of 50000 and 55000 daltons with conservation of the oxygen binding properties of the nondigested protein. We conclude from these studies that the polypeptide chain of Helix pomatia alpha-hemocyanin is folded into about seven compact tertiary structures, which are covalently interconnected. This chain of structural domains has been visualized. (Siezen and Van Bruggen (1974), J. Mol. Biol. 90, 77-89) by electron microscopy, which shows 1/20 hemocyanin molecules to be flexible structures consisting of 7-8 apparently spherical units of 55-60 A diameter.


Thyroid Imaging Reporting and Data System Score Combined with the New Italian Classification for Thyroid Cytology Improves the Clinical Management of Indeterminate Nodules.
The new Italian cytological classification (2014) of thyroid nodules replaced the TIR3 category of the old classification (2007) with two subclasses, TIR3A and TIR3B, with the aim of reducing the rate of surgery for benign diseases. Moreover, thyroid imaging reporting and data system (TI-RADS) score appears to ameliorate the stratification of the malignancy risk. We evaluated whether the new Italian classification has improved diagnostic accuracy and whether its association with TI-RADS score could improve malignancy prediction. We retrospectively analyzed 70 nodules from 70 patients classified as TIR3 according to the old Italian classification who underwent surgery for histological diagnosis. Of these, 51 were available for cytological revision according to the new Italian cytological classification. Risk of malignancy was determined for TIR3A and TIR3B, TI-RADS score, and their combination. A different rate of malignancy (p = 0.0286) between TIR3A (13.04%) and TIR3B (44.44%) was observed. Also TI-RADS score is significantly (p = 0.003) associated with malignancy. By combining cytology and TI-RADS score, patients could be divided into three groups with low (8.3%), intermediate (21.4%), and high (80%) risk of malignancy. In conclusion, the new Italian cytological classification has an improved diagnostic accuracy. Interestingly, the combination of cytology and TI-RADS score offers a better stratification of the malignancy risk.


Safety Profile of Oral Iron Chelator Deferiprone in Chinese Children with Transfusion-Dependent Thalassaemia.
There is a lack of knowledge regarding the incidence of serious adverse drug reactions (ADR) to the oral iron chelator deferiprone in Chinese children with transfusion-dependent thalassaemia. In this retrospective population-based cohort study, paediatric thalassaemia patients in Hong Kong were screened for serious and medically important adverse events related to deferiprone therapy using diagnosis codes, laboratory data and hospital admissions. Potential ADRs were assessed by reviewing concomitant medications, diagnoses and laboratory data and evaluated using standardised causality assessment. Eighty-seven patients contributing 169.8 person-years were included. Thirty ADRs were identified in 21 patients. Most ADRs (56.0%) occurred in the first three months of therapy. Neutropenia occurred in 11 patients (12.6%; incidence rate 6.5 per 100 patient-years) and severe neutropenia (agranulocytosis) was observed in 5 patients (5.7%, incidence rate 2.9 per 100 patient-years). Other identified ADRs involve severe arthropathy, elevated liver enzymes and mild thrombocytopenia. In conclusion, the safety profile of DFP therapy in Chinese children suffering from transfusion-dependent thalassaemia is in line with previous studies of non-Chinese children. However, unlike previous studies, we observed a relatively high incidence of agranulocytosis and neutropenia in patients with simultaneous combined therapy. Hence close monitoring for white blood cell counts is advised in Chinese children under combined iron chelation therapy. Further prospective clinical and pharmacogenetic studies are required to better evaluate this important safety signal. • Half of the identified ADRs related to deferiprone therapy occurred during the first three months of treatment. • A relatively high incidence of agranulocytosis and neutropenia. Hence close monitoring for white blood cell counts is advised in Chinese children under combined iron chelation therapy.


Factors Associated With Failure of Superior Pubic Ramus Screws.
Examine factors associated with fixation failure in patients treated with superior intramedullary ramus screws. Retrospective. Single, Level 1 trauma center. Unstable pelvic ring fractures amenable fixation that included superior intramedullary ramus screws. Percutaneously inserted intramedullary superior ramus screw fixation of superior pubic ramus (SPR) fractures. Loss of reduction (LOR) of the SPR fracture defined as >2 mm displacement on pelvic radiographs at any time point in follow-up. Two hundred eighty-five fractures in 211 patients (age 44, 95% confidence interval 40.8%-46.4%, 59.3% women, 55.1% retrograde screws) were included in the analysis. 14 (4.9%) of fractures had LOR. Patients were significantly more likely to have LOR as age increased (P = 0.01), body mass index (BMI) increased (P = 0.01), and if they were women (P < 0.01). There was a significantly decreased LOR (P < 0.01) as fractures moved further from the pubis symphysis. Retrograde screws were significantly (P < 0.01) more likely to have LOR. In SPR fractures treated with retrograde screws, failure was significantly associated with increasing BMI (P = 0.02), the presence of an inferior ramus fracture (P = 0.02), and trended toward significance with increasing age (P = 0.06), and decreased distance from the symphysis (P = 0.07). Superior ramus screws are associated with a low failure rate (4.9%), which is lower than previously reported. Retrograde screw insertion, distance from the symphysis, increasing age, increasing BMI, decreased distance from the symphysis, and ipsilateral inferior ramus fractures were predictors of failure. In these patients, alternative modalities should be considered, although low rates of failure can still be expected. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Brostallicin (PNU-166196)--a new DNA minor groove binder that retains sensitivity in DNA mismatch repair-deficient tumour cells.
Defects in DNA mismatch repair (MMR) are associated with a predisposition to tumorigenesis and with drug resistance owing to high mutation rates and failure to engage DNA-damage-induced apoptosis. DNA minor groove binders (MGBs) are a class of anticancer agents highly effective in a variety of human cancers. Owing to their mode of action, DNA MGB-induced DNA damage may be a substrate for DNA MMR. This study was aimed at investigating the effect of loss of MMR on the sensitivity to brostallicin (PNU-166196), a novel synthetic alpha-bromoacrylic, second-generation DNA MGB currently in Phase II clinical trials and structurally related to distamycin A. Brostallicin activity was compared to a benzoyl mustard derivative of distamycin A (tallimustine). We report that the sensitivities of MLH1-deficient and -proficient HCT116 human colon carcinoma cells were comparable after treatment with brostallicin, while tallimustine resulted in a three times lower cytotoxicity in MLH1-deficient than in -proficient cells. MSH2-deficient HEC59 parental endometrial adenocarcinoma cells were as sensitive as the proficient HEC59+ch2 cells after brostallicin treatment, but were 1.8-fold resistant after tallimustine treatment as compared to the MSH2-proficient HEC59+ch2 counterpart. In addition, p53-deficient mouse fibroblasts lacking PMS2 were as sensitive to brostallicin as PMS2-proficient cells, but were 1.6-fold resistant to tallimustine. Loss of neither ATM nor DNA-PK affected sensitivity to brostallicin in p53-deficient mouse embryonic fibroblasts, indicating that brostallicin-induced cytotoxicity in a p53-deficient genetic background does not seem to require these kinases. These data show that, unlike other DNA MGBs, MMR-deficient cells retain their sensitivity to this new alpha-bromoacrylic derivative, indicating that brostallicin-induced cytotoxicity does not depend on functional DNA MMR. Since DNA MMR deficiency is common in numerous types of tumours, brostallicin potentially offers the advantage of being effective against MMR-defective tumours that are refractory to several anticancer agents.


In vitro and in vivo hematopoietic effect of mutant human granulocyte colony-stimulating factor.
About 100 derivatives of human recombinant granulocyte colony-stimulating factor (rhG-CSF) were created by various gene-mutagenic techniques, and KW-2228, in which amino acids were replaced at five positions of N-terminal region of intact rhG-CSF, was picked up and evaluated for its biologic and physicochemical properties in comparison with intact rhG-CSF. KW-2228 showed two to four times higher specific activity than that of intact rhG-CSF in mouse and/or human bone marrow progenitor cells by colony-forming unit assay in soft agar, and by cell-proliferation assay in liquid culture. KW-2228 showed a potency to increase peripheral neutrophil counts when it was administered to normal C3H/He mice by single intravenous injection. Increase of total leukocyte count and neutrophils was observed, with peak level at 8 to 12 hours at low doses (0.5 to 1.0 micrograms/mouse), and the highest level was maintained for 24 to 30 hours at high doses (5 to 10 micrograms/mouse). The granulopoietic effect of KW-2228 was examined by several doses of single course (once daily for 10 days) or multiple courses (twice daily injection for 5 days followed by cessation for 9 days on one cycle, 3 cycles in total) of treatment. KW-2228 showed higher activity than that of rhG-CSF, especially at sub-optimal doses of multiple courses of treatment. Furthermore, KW-2228 was found to be more stable physicochemically and biologically than intact rhG-CSF, especially under thermal conditions at 56 degrees C and in the human plasma at 37 degrees C, suggesting a protease resistancy. Pharmacokinetic study showed that plasma concentration of KW-2228 assayed for its bioactivity maintained a higher level than that of intact rhG-CSF for 60 minutes after intravenous injection of this protein to normal mice. Those results suggest that KW-2228 might show a superior in vivo hematopoietic effect to intact rhG-CSF due to its high specific activity to progenitor cells, and also due to its improved physicochemical, biologic, and pharmacokinetic stability in host animals.


[Stages in coping after stroke].
People who suffer a stroke most often experience a major crisis in their lives. This leads to psychological problems which are likely to influence the patient's life satisfaction, long-term rehabilitation outcome, and quality of life as much as the physical consequences caused by the stroke, The patient's ability to accept the new reality created by a disability as a part of their future life is therefore essential. This necessitates an individual coping strategy in the center of which is a mourning process, which unfolds in the same way as other experiences of vital threat, e.g. confrontation with one's own dying. Different stages of mourning that can also be found in stroke patients are nonacceptance of the facts, erupting emotions, parting with the former and finding of a new self as well as new perspectives for one's future. Whether a coping process is successful does not depend on the type of the defense mechanisms or stages, but essentially on their persistence. It is the task of the therapeutical team to support this process by applying a basic therapeutic behavior, stage-consistent interventions and a positive feedback. A failed process of coping may be one of the causes of the high prevalence of psychological disorders like depression and anxiety disorders in patients having suffered a stroke.


Addressing multigenerational conflict: mutual respect and carefronting as strategy.
This article addresses the challenges faced by nurses as they work side-by-side with nurses from a variety of generational cohorts. First a brief overview of the generational characteristics of the four generational cohorts in today's workplace is presented. Next the importance of each nurse using respect and carefronting as antidotes to generational conflict is discussed. Finally the role of nursing leadership in facilitating respect and carefronting is noted.


Association of smoking cigarettes, age, and sex with serum concentrations of olanzapine in patients with schizophrenia.
Olanzapine is an atypical antipsychotic drug which is effective in the treatment of schizophrenia. Cigarette smoking, age, and sex could be related to the pharmacokinetics and serum concentrations of olanzapine in patients with schizophrenia. The aim of the study was to examine whether there was a significant difference in the serum olanzapine concentrations with regard to the mentioned factors. A total of 58 outpatients with schizophrenia (37 smokers, 42 men, 35 older than 40 years) participated in the study. Blood was sampled in serum tubes just before taking the next dose of olanzapine. Olanzapine was extracted by liquid-liquid extraction and was measured by an in-house high-performance liquid chromatography method on Shimadzu Prominence HPLC System with diode array detector SPD-M20A (Shimadzu, Kyoto, Japan). The results were expressed as the ratio of concentration to the daily dose of olanzapine (C/D). Non-parametric statistical tests were used to analyse differences between variables. The median C/D of olanzapine (interquartile range) in smokers was 6.0 (3.4-10.2) nmol/L/mg and in non-smokers 10.1 (5.9-17.6) nmol/L/mg; P = 0.007. The median C/D of olanzapine in patients younger than 40 years was 5.6 (4.5-10.2) nmol/L/mg and in patients older than 40 years 8.4 (5.6-13.0) nmol/L/mg; P = 0.105. The median C/D of olanzapine in male patients was 6.6 (4.6-10.4) nmol/L/mg and in female patients 9.0 (5.9-15.3) nmol/L/mg; P = 0.064. The serum olanzapine concentration was significantly lower in smoking than in non-smoking patients with schizophrenia. No significant difference was demonstrated with regard to age and sex.


Summer fescue toxicity in dairy steers fed tall fescue seed.
Seeds of the tall fescue ( Festuca arundinacea Schreb .) cultivars Kentucky 31 and an experimental ryegrass X tall fescue hybrid derivative strain (G1-307), and orchard grass (OG) seed were fed in a carrier diet to calves in controlled environmental rooms (31 to 32 C). Both tall fescue varieties produced symptoms of summer toxicosis in dairy steers. Total feed intake (P less than .01) and water intake (P less than .01) of calves were reduced by the tall fescue seed diets when compared with orchard grass. Steers fed G1-307 and Kentucky 31 tall fescue lost (NS) 17.5 and 7.8 kg of body weight, respectively, while those consuming orchard grass gained (P less than .01) 6.2 kg during the experiment. Rectal temperatures were lower (P less than .05) in the calves fed OG (39.4 C) when compared with those fed G1-307 (40.6 C) and Kentucky 31 (40.8 C) tall fescue seed, respectively. In a second trial Kentucky 31 seed was fed in a carrier diet to Holstein steers at graded levels of 0, 350, 700 and 1,050 g seed/d. Consumption of 700 and 1,050 g seed/d adversely affected performance of steers. Total feed intake was lower and water intake was reduced (P less than .05), with rectal temperatures being elevated (P less than .01) in these two groups when compared with 0- or 350-g treatments. No significant differences were detected in body weight changes in this trial. Respiration rates were not significantly affected in either trial. Results of these trials with the young bovine indicate that a toxic substance(s) is present in tall fescue seed.


Assessing cross-reactivity of Junín virus-directed neutralizing antibodies.
Arenaviruses cause several viral hemorrhagic fevers endemic to Africa and South America. The respective causative agents are classified as biosafety level (BSL) 4 pathogens. Unlike for most other BSL4 agents, for the New World arenavirus Junín virus (JUNV) both a highly effective vaccination (Candid#1) and a post-exposure treatment, based on convalescent plasma transfer, are available. In particular, neutralizing antibodies (nAbs) represent a key protective determinant in JUNV infection, which is supported by the correlation between successful passive antibody therapy and the levels of nAbs administered. Unfortunately, comparable resources for the management of other closely related arenavirus infections are not available. Given the significant challenges inherent in studying BSL4 pathogens, our goal was to first assess the suitability of a JUNV transcription and replication-competent virus-like particle (trVLP) system for measuring virus neutralization under BSL1/2 conditions. Indeed, we could show that infection with JUNV trVLPs is glycoprotein (GP) dependent, that trVLP input has a direct correlation to reporter readout, and that these trVLPs can be neutralized by human serum with kinetics similar to those obtained using authentic virus. These properties make trVLPs suitable for use as a proxy for virus in neutralization assays. Using this platform we then evaluated the potential of JUNV nAbs to cross-neutralize entry mediated by GPs from other arenaviruses using JUNV (strain Romero)-based trVLPs bearing GPs either from other JUNV strains, other closely related New World arenaviruses (e.g. Tacaribe, Machupo, Sabiá), or the distantly related Lassa virus. While nAbs against the JUNV vaccine strain are also active against a range of other JUNV strains, they appear to have little or no capacity to neutralize other arenavirus species, suggesting that therapy with whole plasma directed against another species is unlikely to be successful and that the targeted development of cross-specific monoclonal antibody-based resources is likely needed. Such efforts will be supported by the availability of this BSL1/2 screening platform which provides a rapid and easy means to characterize the potency and reactivity of anti-arenavirus neutralizing antibodies against a range of arenavirus species.


Analysis of primary aromatic amines in the mainstream waterpipe smoke using liquid chromatography-electrospray ionization tandem mass spectrometry.
In recent years waterpipe smoking has spread worldwide and emerged as global health issue. Yet only little is known on the composition of waterpipe smoke. Here, we present a study on the identification and quantification of primary aromatic amines (PAAs) in this complex environmental matrix. Smoking of the waterpipe was conducted with a smoking machine and particulate matter was collected on glass fiber pads. We developed a fast, simple and specific liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) approach to simultaneously detect 31 different PAAs in this matrix. The detection limits comprised a range of 0.45-4.50 ng per smoking session, represented by 2-aminobiphenyl and 3,4,5-trichloroaniline, respectively. Intra- and inter-day precision were determined and proved excellent. We detected 31.3 ± 2.2 ng aniline and 28.0 ± 1.6 ng 4,4'-oxydianiline in the smoke of one waterpipe session. The water in the bowl exerted a small but considerable filter effect on PAAs. The method worked-out showed excellent sensitivity and specificity and is thus highly suited for the determination of PAAs in mainstream waterpipe smoke.


Evaluation of the Hemodynamic Effects of Intravenous Amiodarone Formulations During the Maintenance Phase Infusion.
Two of the excipients in intravenous formulations of amiodarone, polysorbate 80 and benzyl alcohol, have been shown to cause hypotension. A newer formulation of amiodarone, which contains cyclodextrin, is devoid of these excipients. To evaluate the change in mean arterial pressure when utilizing 2 intravenous amiodarone formulations. This was a retrospective cohort analysis conducted at an academic medical center. Patients received intravenous amiodarone containing either polysorbate 80/benzyl alcohol (control) or cyclodextrin (cyclodextrin). Patients received these formulations based on a standard institutional protocol of 1 mg/min for 6 hours, followed by 0.5 mg/min for at least 18 hours or until discontinued by the provider. All data were collected from the medical record and included changes in blood pressures, time to lowest systolic blood pressure, concurrent antihypertensive use, and number of patients requiring treatment for hypotension. A total of 160 patients (120 control, 40 cyclodextrin) were included. There was a statistically significant difference in mean arterial pressure between the groups receiving the control formulation of amiodarone compared with the cyclodextrin formulation across the 24-hour maintenance phase infusion (P < 0.001). There was a significant difference between formulations with regard to the change in mean arterial pressure during the 0- to 6-hour and 12- to 18-hour time blocks. There was a statistically significant difference in the number of patients receiving fluid boluses for treatment of hypotension (P = 0.001). The excipients in the formulation of intravenous amiodarone may have a significant role in the hypotensive effects seen throughout the duration the maintenance phase infusion.