Burden of disease and productivity impact of Streptococcus suis infection in Thailand.
Streptoccocus suis (S.suis) infection is a neglected zoonosis disease in humans mainly affects men of working age. We estimated the health and economic burden of S.suis infection in Thailand in terms of years of life lost, quality-adjusted life years (QALYs) lost, and productivity-adjusted life years (PALYs) lost which is a novel measure that adjusts years of life lived for productivity loss attributable to disease. A decision-analytic Markov model was developed to simulate the impact of S. suis infection and its major complications: death, meningitis and infective endocarditis among Thai people in 2019 with starting age of 51 years. Transition probabilities, and inputs pertaining to costs, utilities and productivity impairment associated with long-term complications were derived from published sources. A lifetime time horizon with follow-up until death or age 100 years was adopted. The simulation was repeated assuming that the cohort had not been infected with S.suis. The differences between the two set of model outputs in years of life, QALYs, and PALYs lived reflected the impact of S.suis infection. An annual discount rate of 3% was applied to both costs and outcomes. One-way sensitivity analyses and Monte Carlo simulation modeling technique using 10,000 iterations were performed to assess the impact of uncertainty in the model. This cohort incurred 769 (95% uncertainty interval [UI]: 695 to 841) years of life lost (14% of predicted years of life lived if infection had not occurred), 826 (95% UI: 588 to 1,098) QALYs lost (21%) and 793 (95%UI: 717 to 867) PALYs (15%) lost. These equated to an average of 2.46 years of life, 2.64 QALYs and 2.54 PALYs lost per person. The loss in PALYs was associated with a loss of 346 (95% UI: 240 to 461) million Thai baht (US$11.3 million) in GDP, which equated to 1.1 million Thai baht (US$ 36,033) lost per person. S.suis infection imposes a significant economic burden both in terms of health and productivity. Further research to investigate the effectiveness of public health awareness programs and disease control interventions should be mandated to provide a clearer picture for decision making in public health strategies and resource allocations.


Rotational dynamics of actin-bound intermediates in the myosin ATPase cycle.
We have used saturation-transfer electron paramagnetic resonance (ST-EPR) to detect the microsecond rotational motions of spin-labeled myosin subfragment one (MSL-S1) bound to actin in the presence of the ATP analogues AMPPNP (5'-adenylylimido diphosphate) and ATP gamma S [adenosine 5'-O-(3-thiotriphosphate)], which are believed to trap myosin in strongly and weakly bound intermediate states of the actomyosin ATPase cycle, respectively. Sedimentation binding measurements were used to determine the fraction of myosin heads bound to actin under ST-EPR conditions and the fraction of heads containing bound nucleotide. ST-EPR spectra were then corrected to obtain the spectrum corresponding to the ternary complex (actin.MSL-S1.nucleotide). The ST-EPR spectrum of MSL-S1.AMPPNP bound to actin is identical to that obtained in the absence of nucleotide (rigor complex), indicating no rotational motion of MSL-S1 relative to actin on the microsecond time scale. However, MSL-S1-ATP gamma S bound to actin is rotationally mobile, with an effective rotational correlation time (tau r) of 17 +/- 2 microseconds. This motion is similar to that observed previously for actin-bound MSL-S1 during the steady-state hydrolysis of ATP [Berger et al. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 8753-8757]. We conclude that, in solution, the weakly bound actin-attached states of the myosin ATPase cycle undergo microsecond rotational motions, while the strongly bound intermediates do not, and that these motions are likely to be involved in the molecular mechanism of muscle contraction.


Expression of the transcription factor Pax 6 in the adult rat dentate gyrus.
The transcription factor Pax 6 is expressed in precursor cells during embryonic CNS development, and it plays an important role in the regulation of cell proliferation and neuronal fate determination. Pax 6-expressing cells are also present in the adult hippocampal dentate gyrus and subventricular zone/rostral migratory stream, regions in which neuronal precursors exist during adult life. In the adult dentate gyrus, precursor cells are located in the innermost portion of the granule cell layer, and Pax 6-expressing nuclei are most abundant in this region. To examine the putative role of Pax 6 in adult hippocampal neurogenesis, we have studied the proliferative activity, distribution, and phenotype of Pax 6-expressing cells by using immunohistochemistry. Our results indicate that Pax 6 is intensely expressed in proliferating precursors of the adult dentate gyrus. Pax 6 is also expressed in nonproliferating cells, which may correspond to resting progenitor cells and to granule neurons in their very early developmental stages, because this transcription factor is strongly down-regulated during granule neuron differentiation. However, a small subpopulation of hilar mature neurons and certain astrocytes of the adult hippocampus also express Pax 6. Although the precise roles of this transcription factor in the adult brain remain to be determined, our findings support the idea that its function in the control of cell proliferation and neuronal fate determination during embryogenesis is also operative in the adult hippocampus. However, the expression of Pax 6 in astrocytes and certain mature neurons may indicate the existence of other roles for this transcription factor in this telencephalic region.


Pedigree and marker information requirements to monitor genetic variability.
There are several measures available to describe the genetic variability of populations. The average inbreeding coefficient of a population based on pedigree information is a frequently chosen option. Due to the developments in molecular genetics it is also possible to calculate inbreeding coefficients based on genetic marker information. A simulation study was carried out involving ten sires and 50 dams. The animals were mated over a period of 20 discrete generations. The population size was kept constant. Different situations with regard to the level of polymorphism and initial allele frequencies and mating scheme (random mating, avoidance of full sib mating, avoidance of full sib and half sib mating) were considered. Pedigree inbreeding coefficients of the last generation using full pedigree or 10, 5 and 2 generations of the pedigree were calculated. Marker inbreeding coefficients based on different sets of microsatellite loci were also investigated. Under random mating, pedigree-inbreeding coefficients are clearly more closely related to true autozygosity (i.e., the actual proportion of loci with alleles identical by descent) than marker-inbreeding coefficients. If mating is not random, the demands on the quality and quantity of pedigree records increase. Greater attention must be paid to the correct parentage of the animals.


The intra- and interobserver variability of impedance cardiography in patients at rest and during exercise.
We studied the intra- and interobserver variability in the calculation of stroke volume by the impedance technique, using the recently proposed refinements in the electrode configuration and signal processing. Three groups of patients were included in this study: ten stable cardiac patients who underwent a diagnostic heart catheterization, ten patients 24-26 h after coronary artery bypass surgery and ten patients with severe chronic obstructive pulmonary disease (COPD). The first two groups were studied at rest and the COPD group during submaximal exercise. The intra-observer variability was 4.2%, 3.9-4.0% and 6.0-6.9% for the catheterized, surgical and COPD groups, respectively. The interobserver variability was 4.3%, 2.6% and 2.4%, respectively. It is concluded that highly reproducible data can be obtained with the newly proposed impedance technique in patients at rest and exercise which may be comparable or superior to other techniques used in clinical settings.


Development and Evaluation of Portal do Zumbido : A Novel Telehealth Approach to Tinnitus.
Introduction Tinnitus is a prevalent symptom in audiological clinics, and the speech therapist participates in the assessment, diagnosis, and intervention of tinnitus. A range of studies have been developed with the aim to promote auditory health, and telehealth is an option for this purpose. Objective The aim of the present study was to develop and assess a virtual platform for tinnitus management for speech therapists. Methods The present study provided a distance-learning-course, with the aid of Portal do Zumbido , available in the following electronic address: zumbido.fob.usp.br. Infographics were available for download, and modules were made to comprise the introduction to the symptoms, assessment, and intervention for tinnitus. Results Speech therapists of different regions of Brazil received an e-mail invitation to participate and to assess the platform, 46 of whom registered. From the 46 speech therapists registered, 38 assessed the virtual platform with the aid of a questionnaire about the theoretical content and then answered a motivational research sheet (MRS). The speech therapists correctly answered 90% and 100% of the questions in the questionnaire and in the MRS, respectively, which may indicate that the professionals positively absorbed the content; thus, the content was considered to be impressive when the MRS was analyzed. Conclusion Portal do Zumbido was developed and is available for access in the following electronic address: zumbido.fob.usp.br. Furthermore, the platform received a positive assessment by the speech therapists who participated in the present study.


Persistence of Epigenomic Effects After Recovery From Repeated Treatment With Two Nephrocarcinogens.
The discovery of the epigenetic regulation of transcription has provided a new source of mechanistic understanding to long lasting effects of chemicals. However, this information is still seldom exploited in a toxicological context and studies of chemical effect after washout remain rare. Here we studied the effects of two nephrocarcinogens on the human proximal tubule cell line RPTEC/TERT1 using high-content mRNA microarrays coupled with miRNA, histone acetylation (HA) and DNA methylation (DM) arrays and metabolomics during a 5-day repeat-dose exposure and 3 days after washout. The mycotoxin ochratoxin A (OTA) was chosen as a model compound for its known impact on HA and DM. The foremost effect observed was the modulation of thousands of mRNAs and histones by OTA during and after exposure. In comparison, the oxidant potassium bromate (KBrO3) had a milder impact on gene expression and epigenetics. However, there was no strong correlation between epigenetic modifications and mRNA changes with OTA while with KBrO3 the gene expression data correlated better with HA for both up- and down-regulated genes. Even when focusing on the genes with persistent epigenetic modifications after washout, only half were coupled to matching changes in gene expression induced by OTA, suggesting that while OTA causes a major effect on the two epigenetic mechanisms studied, these alone cannot explain its impact on gene expression. Mechanistic analysis confirmed the known activation of Nrf2 and p53 by KBrO3, while OTA inhibited most of the same genes, and genes involved in the unfolded protein response. A few miRNAs could be linked to these effects of OTA, albeit without clear contribution of epigenetics to the modulation of the pathways at large. Metabolomics revealed disturbances in amino acid balance, energy catabolism, nucleotide metabolism and polyamine metabolism with both chemicals. In conclusion, the large impact of OTA on transcription was confirmed at the mRNA level but also with two high-content epigenomic methodologies. Transcriptomic data confirmed the previously reported activation (by KBrO3) and inhibition (by OTA) of protective pathways. However, the integration of omic datasets suggested that HA and DM were not driving forces in the gene expression changes induced by either chemical.


Management of a Trichophyton tonsurans outbreak in a day-care center.
Trichophyton tonsurans is the leading cause of tinea capitis in the United Kingdom (UK) as well as causing tinea corporis. This organism has been linked to several outbreaks in the UK and abroad, and such outbreaks may be prolonged since T. tonsurans can be difficult to control. There remains an incomplete consensus in the literature on the optimal management of such outbreaks of this infection. Following notification that a child with T. tonsurans was identified at a day-care center in the UK, initial investigations identified nine cases of fungal infection involving children and staff over the previous 7 months. We report on the management of an outbreak of T. tonsurans tinea capitis and tinea corporis among children and staff in a day-care center. An outbreak control team with representatives from dermatology, microbiology, day-care center management, and the Health Protection Agency initiated case ascertainment by scalp inspection and brushing of all children and staff at the nursery. Two complete rounds of screening were required before the outbreak was declared over. Infection control measures included antifungal shampoo use, exclusion of identified cases for a short period, removal of shared items from the center, and enhanced decontamination of fomites. The outbreak, which lasted longer than 12 months, involved 12 children and 7 staff members. Of these, 12 cases were confirmed by positive fungal culture. T. tonsurans is difficult to manage, especially in childcare settings, but case ascertainment, appropriate treatment with oral agents, and sustained infection control measures can be effective in controlling such outbreaks.


The C-terminal proline-rich tail of human immunodeficiency virus type 2 Vpx is necessary for nuclear localization of the viral preintegration complex in nondividing cells.
Human immunodeficiency virus type 2 (HIV-2), like other lentiviruses, is capable of infecting nondividing T cells and macrophages. The present work shows that in HIV-2-infected cells, Vpx is necessary for efficient nuclear import of the preintegration complex. In agreement with this finding, the subcellular localization of a GFP-Vpx fusion protein was found to be predominantly nuclear. However, deletion of the proline-rich C-terminal 11 residues of Vpx resulted in a shift of the fusion protein to the cytoplasm. Furthermore, the same deletion in the context of the provirus resulted in a decrease in nuclear import of the preintegration complex and attenuated replication in macrophages.


Flow cytometric characterization of cecal appendix lymphocyte subpopulations in children: a pilot study.
Scientific literature regarding the characterization of lymphocyte subpopulations of the cecal appendix is sparse, with few precedents limited to immunohistochemical techniques. We conducted a prospective pilot study to characterize lymphocyte subpopulations of the cecal appendix in children. Participants were divided into three groups: (1) patients without histological acute appendiceal inflammation, (2) patients with histological uncomplicated acute appendicitis, and (3) patients with histological complicated acute appendicitis (gangrenous, perforated). A fresh sample of the base of the appendix was taken from all patients and a flow cytometric study was performed. Quantitative variables were compared using Kruskal-Wallis test and Mann-Whitney U test. This study included 57 patients divided into Group 1 (n = 5), Group 2 (n = 37), and Group 3 (n = 15). Median values (IQR) of the percentage of B-lymphocytes were 67.8 [66.8-68.1] in group 1, 61.15 [53.74-66.4] in group 2, and 52.1 [33-62.02] in group 3 (p = 0.02). Median values (IQR) of the percentage of NK-lymphocytes were 0.26 [0.2-0.3] in group 1, 0.55 [0.37-0.66] in group 2, and 0.84 [0.35-1.45] in group 3 (p = 0.008). Median values (IQR) of the percentage of T-lymphocytes were 31.9 [31.7-33.1] in group 1, 37.68 [32.15-45.69] in group 2, and 46.9 [37.03-67] in group 3 (p = 0.02). Pair comparisons of groups 2 and 3 also showed significant differences in the percentage of B lymphocytes (p = 0.03) and NK-lymphocytes (p = 0.02). Significant differences in lymphocyte subpopulations were identified according to the histologic grade of the cecal appendix. More specifically, a lower percentage of B-lymphocytes and a higher percentage of T- and NK-lymphocytes were observed in cases of acute appendicitis. These findings must be confirmed and their etiopathogenic, diagnostic, and prognostic implications elucidated in future studies with larger sample sizes.


Digital financial inclusion and welfare: Effect, mechanism and imbalance.
This study utilizes provincial panel data from China spanning from 2013 to 2019 and considers indices such as residents' income, employment, housing, education, and medical care to comprehensively measure the welfare index of each province using the coefficient of variation. It also examines the impact and mechanism of digital financial inclusion on welfare and evaluates its influence on welfare imbalance through panel data models and RIF regression. The estimation results of digital financial inclusion on welfare indicate that digital financial inclusion is an important driver elevating the income, employment, housing, education, medical treatment and other welfares of Chinese residents, and the conclusion remains robust after using instrumental variable estimation and replacing the measurement method of welfare. Regional differences reveal that digital financial inclusion has a greater contribution to the welfare in the Midwest. The effect mechanism test shows that digital financial inclusion improves the welfare by creating employment opportunities and plays a greater role in the Midwest, and will also improve the welfare by increasing the welfare expenditure. However, this mechanism will exacerbate the welfare imbalance between the Midwest and East. The estimation results of digital financial inclusion on the welfare imbalance show that digital financial inclusion reduces welfare imbalance between the central and western regions and the eastern regions, as well as within the central and western regions.


Biosimilars: a regulatory perspective from America.
Biosimilars are protein products that are sufficiently similar to a biopharmaceutical already approved by a regulatory agency. Several biotechnology companies and generic drug manufacturers in Asia and Europe are developing biosimilars of tumor necrosis factor inhibitors and rituximab. A biosimilar etanercept is already being marketed in Colombia and China. In the US, several natural source products and recombinant proteins have been approved as generic drugs under Section 505(b)(2) of the Food, Drug, and Cosmetic Act. However, because the complexity of large biopharmaceuticals makes it difficult to demonstrate that a biosimilar is structurally identical to an already approved biopharmaceutical, this Act does not apply to biosimilars of large biopharmaceuticals. Section 7002 of the Patient Protection and Affordable Care Act of 2010, which is referred to as the Biologics Price Competition and Innovation Act of 2009, amends Section 351 of the Public Health Service Act to create an abbreviated pathway that permits a biosimilar to be evaluated by comparing it with only a single reference biological product. This paper reviews the processes for approval of biosimilars in the US and the European Union and highlights recent changes in federal regulations governing the approval of biosimilars in the US.


"Our Job is that Whole Gray Zone in Between There": Investigating Genetic Counselors' Strategies for Managing and Communicating Uncertainty.
Ample research suggests that uncertainty is a major concern for individuals undergoing genetic testing and for genetic mutation carriers, and that their management of uncertainty is largely influenced by healthcare providers, including genetic counselors (GCs). Indeed, uncertainty is inherent in genetic testing results. To help patients grapple with uncertainty, GCs need to effectively manage the uncertainty inherent in genetic testing and communicate it appropriately to patients. The present study aimed to identify GCs' strategies for managing uncertainty and communicating uncertainty to patients. Eighteen GCs participated in five focus group interviews, and the data were analyzed using a grounded theory approach. Findings indicated that GCs used three main strategies to manage uncertainty: becoming lifelong learners, normalizing uncertainty, and seeking social support. To communicate uncertainty to patients, GCs also used three different strategies: engaging in open and honest communication, adapting to patients' needs, and focusing on known information. The implications of these findings for uncertainty theories and for conceptual frameworks of GCs' clinical communication are discussed.


Connecting, learning, supporting: Caregivers' experiences of a stress and distress biopsychosocial group intervention.
Family caregivers are fundamental in supporting people living with dementia to remain at home, however, psychological distress can occur as a result of their caring role. Research into interventions for caregivers of people living with young-onset dementia, including their experience of and the mediating processes of such interventions, remains limited. An Interpretative Phenomenological Analysis explored caregiver experiences and influence on caregiving of participating in a "Responding to Distress in Dementia" group. Five family caregivers were interviewed with discussions covering the period from first noticing symptoms to the interview session. Within the group experience, four superordinate themes were identified: 'connecting to other caregivers', 'learning about caregiving', 'group factors' and 'reduced caregiver distress'. During the post-group period, three superordinate themes were recognised: 'maintaining support', 'applying learning', and 'normalising caregiving'. The study highlighted several interrelated themes involving creating connections amongst caregivers with similar experiences, social learning, and supportive learning through group structure and facilitation. Many of the processes reflected those found in existing dementia caregiver intervention research. Recommendations included facilitating peer support groups and exploring whole-family approaches.


The dark side of testosterone deficiency: II. Type 2 diabetes and insulin resistance.
A considerable body of evidence exists suggesting a link among reduced testosterone plasma levels, type 2 diabetes (T2D), and insulin resistance (IR). Hypogonadal men are at higher risk for T2D. Here we evaluate the relationships between testosterone, metabolic syndrome (MetS), T2D, and IR and discuss the relationships among androgen deficiency and these factors, especially as it ultimately relates to the development of cardiovascular disease and erectile dysfunction (ED). Thus, a comprehensive literature search was carried out using PubMed, and relevant articles pertinent to androgen deficiency, T2D, IR, MetS, and ED were reviewed and discussed. Low testosterone precedes elevated fasting insulin, glucose, and hemoglobin A1c (HbA1C) values and may even predict the onset of diabetes. Treatment of prostate cancer patients with surgical or medical castration exacerbates IR and glycemic control, strengthening the link between testosterone deficiency and onset of T2D and IR. Androgen therapy of hypogonadal men improves insulin sensitivity, fasting glucose, and HbA1c levels. We suggest that androgen deficiency is associated with IR, T2D, MetS, and with increased deposition of visceral fat, which serves as an endocrine organ, producing inflammatory cytokines and thus promoting endothelial dysfunction and vascular disease.


Genetic basis of salt-susceptibility in the Sabra rat model of hypertension.
The Sabra salt-sensitive SBH/y and salt-resistant SBN/y rats constitute a unique experimental model of hypertension in which salt-susceptibility is genetically determined and expressed only after salt-loading, without the development of spontaneous hypertension. To determine the genetic basis of salt-susceptibility in the Sabra rats, the candidate gene and total genome screen approaches were adopted. The likely candidate genes in this model incorporate salt-related physiological mechanisms such as the nitric oxide system, the arginine vasopressin axis and the epithelial sodium channel. In the random genome search scheme for culprit genes, SBH/y and SBN/y were cross-bred. A highly unusual and composite mode of transmission of salt-susceptibility was found in this cross, emphasizing the complexity of the genetic basis of salt-susceptibility. Linkage analysis of the entire rat genome with a large number of widely distributed microsatellite markers identified three putative gene loci on chromosomes 1 and 17 that contribute importantly to salt-sensitivity and/or resistance, and uncovered sex specificity in the role that salt-susceptibility genes fulfill in the development of hypertension.


Preservation of 32 human kidneys by simple cold storage for more than 48 hours.
Transplant centers are reluctant to use kidneys stored cold for more than 48 hours. During a 6-year interval we transplanted 32 kidneys preserved by intracellular electrolyte flushing that were stored cold for 48.2 to 61.4 hours. Of the recipients 91 per cent required dialysis within 1 week after transplantation. The mean serum creatinine nadir within 1 month was 3.0 mg. per dl. and graft survival at 1 month was 81 per cent. Short-term kidney graft function was not influenced significantly by the addition of magnesium sulfate to the flush solutions or by cyclosporin immunosuppression. The 1 and 2-year actuarial kidney graft survival rates were 72 and 58 per cent, respectively. The 1 and 2-year mean serum creatinine levels were 1.9 and 1.6 mg. per dl., respectively. Kidneys can be transplanted successfully after 48 hours of simple cold storage following flushing with an ice-cold intracellular electrolyte solution.


Toxicological evaluation of polyphenol extract from Acerola (Malpighia emarginata DC.) fruit.
Acerola (Malpighia emarginata DC.) is known to be one of the best fruits rich in ascorbic acid and polyphenols. Acerola polyphenols (APs) were found to have radical scavenging activities and inhibitory effects on both alpha-glucosidase and advanced glycation endproducts (AGEs) production. While polyphenols from several foodstuffs have been shown to be safe, APs, which differ from those of other foodstuffs, have not been evaluated for their toxicological properties. Here, we report preliminary toxicological and safety evaluations of crude APs (C-AP), which were obtained by eluting an XAD7HP column-adsorbed fraction of APs with 70% ethanol containing malic acid. The total polyphenol content of C-AP was 57.7% with the main polyphenols being proanthocyanidin and cyanidin-3-alpha-O-rhamnoside. For toxicological evaluations, C-AP was administered orally to rats at doses of 2000 mg/kg body weight (acute) or 100, 300, and 1000 mg/kg body weight/d for 28 (subacute) and 90 (subchronic) d. In the acute oral toxicological test, no deaths or abnormalities at necropsy on day 14 were observed, confirming that the minimum fatal dose of C-AP is greater than 2000 mg/kg body weight. In both subacute and subchronic toxicological tests, no death was recorded and the body weights and food intakes of the rats did not differ significantly from the control groups. Besides, there were no abnormal clinical signs related to administration of C-AP in any of the experimental animals. These results provide an important reference for the safety of APs as a food supplement for human consumption.


Impaired glucose metabolism and type 2 diabetes are associated with hypercoagulability: potential role of central adiposity and low-grade inflammation--the Hoorn Study.
Type 2 diabetes (DM2) is associated with greater risk for cardiovascular disease (CVD), which may, at least partially, be explained by prothrombotic alterations. We therefore investigated; first, the extent to which individuals with impaired glucose metabolism (IGM) and/or DM2 had greater levels of thrombin generation than those with normal glucose metabolism (NGM); and second, whether any differences were independent of other cardiovascular risk factors, such as smoking, hypertension, dyslipidaemia, (micro)albuminuria, glycemic control and (central) adiposity, and/or were potentially 'mediated' by low-grade inflammation (high-sensitivity C-reactive protein (hsCRP)). We studied 744 individuals from the Hoorn Study (275 NGM, 176 IGM and 293 DM2, mean age 68.6 ± 7.1 years). Thrombin generation in platelet-poor plasma was measured using the Calibrated Automated Thrombogram and three parameters were derived: lag time, peak height and endogenous thrombin potential (ETP). Data were analyzed with multiple linear regression analyses. After adjustment for age, sex, prior CVD and smoking status, individuals with IGM or DM2 had a longer lag time [ß = 0.14 min (95% CI: 0.02; 0.26)], higher peak height [ß = 7.29 nM (-1.33; 15.91)] and ETP [ß = 35.65 nM*min (0.97; 70.34)] than those with NGM. These differences were attenuated to ß = 0.06 min (-0.07; 0.19), 3.82 nM (-5.46; 13.10) and 16.34 nM*min (-20.92; 53.59), respectively, when further adjusted for waist circumference and hsCRP. Individuals with IGM or DM2 had up to 4% higher thrombin generation compared with NGM, which may be explained, to a great extent, by the greater levels of central adiposity and related low-grade inflammation characterizing these individuals.


Study of a manual method of liquid-based cervical cytology.
We report a study of a manual liquid-based cytology (MLBC) method. Slides are prepared by using a polymer solution and allowing it to dry, forming a membrane. The aims of the study were to prepare cervical cytology smears using the manual method, observe morphology, compare with direct scrape smears and correlate with histopathology wherever possible. Out of 105 cases, the membrane was intact, indicating a good MLBC preparation, in 97 cases. Simultaneous conventional smears were taken in 81 patients. There was an 88.8% agreement in the diagnoses of general category in both groups. The diagnosis of negative for intraepithelial lesion or malignancy (NILM) in both groups was made in 70 cases. The MLBC preparation was unsatisfactory in two cases which showed high grade squamous intraepithelial lesion and low grade squamous intraepithelial lesion, respectively, on the conventional smear. One MLBC smear diagnosed as atypical squamous cells of undetermined significance was reported as NILM on conventional smear. Cytohistologic correlation was done in nine cases, all of which showed cervicitis on histopathology. The MLBC method was found to be comparable to the conventional scrape smear. Further study of this method as a cost-effective alternative to the mechanized methods would be worthwhile.


Frontal cortex functioning in the infant broader autism phenotype.
Atypical attention has been proposed as a marker of the broader autism phenotype. In the present study we investigated this and the related process of inhibitory control at the youngest possible age through the study of infant siblings of children with an autism spectrum disorder (Sibs-ASD). Both attention and inhibition have been related to the frontal cortex of the brain. Nine- to ten-month-old Sibs-ASD and low-risk control infants completed the Freeze-Frame task, in which infants are encouraged to inhibit looks to peripherally presented distractors whilst looking at a central animation. The attractiveness of the central stimulus is varied in order to investigate the selectivity of infants' responses. In line with previous studies, it was found that a subset of Sibs-ASD infants had difficulty disengaging attention from a central stimulus in order to orient to a peripheral stimulus. The Sibs-ASD group also showed less Selective Inhibition than controls. However, Sibs-ASD infants did demonstrate Selective Inhibitory Learning. These results provide preliminary evidence for atypical frontal cortex functioning in the infant broader autism phenotype.


Degradation and induction specificity in actinomycetes that degrade p-nitrophenol.
We have isolated two soil bacteria (identified as Arthrobacter aurescens TW17 and Nocardia sp. strain TW2) capable of degrading p-nitrophenol (PNP) and numerous other phenolic compounds. A. aurescens TW17 contains a large plasmid which correlated with the PNP degradation phenotype. Degradation of PNP by A. aurescens TW17 was induced by preexposure to PNP, 4-nitrocatechol, 3-methyl-4-nitrophenol, or m-nitrophenol, whereas PNP degradation by Nocardia sp. strain TW2 was induced by PNP, 4-nitrocatechol, phenol, p-cresol, or m-nitrophenol. A. aurescens TW17 initially degraded PNP to hydroquinone and nitrite. Nocardia sp. strain TW2 initially converted PNP to hydroquinone or 4-nitrocatechol, depending upon the inducing compound.


Identification of a SIRT1 mutation in a family with type 1 diabetes.
Type 1 diabetes is caused by autoimmune-mediated β cell destruction leading to insulin deficiency. The histone deacetylase SIRT1 plays an essential role in modulating several age-related diseases. Here we describe a family carrying a mutation in the SIRT1 gene, in which all five affected members developed an autoimmune disorder: four developed type 1 diabetes, and one developed ulcerative colitis. Initially, a 26-year-old man was diagnosed with the typical features of type 1 diabetes, including lean body mass, autoantibodies, T cell reactivity to β cell antigens, and a rapid dependence on insulin. Direct and exome sequencing identified the presence of a T-to-C exchange in exon 1 of SIRT1, corresponding to a leucine-to-proline mutation at residue 107. Expression of SIRT1-L107P in insulin-producing cells resulted in overproduction of nitric oxide, cytokines, and chemokines. These observations identify a role for SIRT1 in human autoimmunity and unveil a monogenic form of type 1 diabetes.


Are allogeneic and associative recognition coincident T cell paradigms?
T lymphocytes are obsessed with cell surface molecules encoded by genes of the MHC, particularly class I (K/D) gene products. In one paradigm, T cells recognize and are activated by allodisparate class I antigens (allogeneic recognition); in another paradigm, T lymphocytes recognize viral antigens or haptens in the context of syngeneic class I determinants (associative recognition). Whether these paradigms are coincident, or even overlapping, is unresolved. We have studied neonatal tolerance induction among the H-2Kb parent strain and several of its class I mutants as a means of analyzing the fine discrimination potential of T cell recognition. Neonatal C57BL/6 animals were rendered tolerant of Kb mutant antigens; bm1, bm3, bm5, or bm8. Similar panels of mutant mice were rendered tolerant neonatally of the parent Kb antigen. When fully tolerant, mature animals were challenged with third-party grafts bearing related Kb alloantigenic disparities, they proved to be highly discriminant. Kb animals tolerant of bm1 or bm3 always rejected third-party grafts bearing bm3 and bm1 antigens, respectively. Similarly, the majority of Kb animals tolerant of bm5 or bm8 (and vice versa) were able to distinguish accurately the tolerated antigen from third-party antigens, rejecting grafts bearing the latter. Since associative recognition involving these antigens has been found to be at least partially degenerate and since neonatally tolerant animals nonetheless remain highly discriminant of these same antigenic variations, it is tempting to conclude that associative and allogeneic recognition address different aspects of T cell function.


A new colorimetric and fluorescent probe with a large stokes shift for rapid and specific detection of biothiols and its application in living cells.
In this work, a new reversible colorimetric and fluorescent probe for sequential recognition of copper ions and biothiols is synthesized easily. Based on the chelation-enhanced fluorescence quenching (CHEQ) effect, this probe shows high sensitivity and selectivity towards Cu2+, which can be detected by the naked eye. And this experimental phenomenon can also be realised upon addition of biothiols, restoring their initial fluorescence intensity. This probe also features a very high response speed (less than 5 seconds) and a large stokes shift (178 nm) toward Cu2+ and biothiols. Moreover, the detection limit for Cu2+ and biothiols is as low as 7.34 nM and 10.3 nM, respectively. Additionally, this ON-OFF-ON-type fluorescence recognition cycle can be repeated more than 5 times by addition of Cu2+ and biothiols in turn. Particularly, this 1-Cu2+ ensemble is further successfully applied for GSH detection in living cells.


Control of the number of cell division rounds in distinct tissues during ascidian embryogenesis.
An important question in developmental biology is how the number of cells constituting the body is regulated during development. After termination of cell divisions, cells enter the postmitotic phase, but the mechanisms determining the timing of final cell division remain unclear. In ascidians, the egg develops into a tadpole larva consisting of a relatively small number of cells (approximately 2800), indicating that embryonic cells divide 11.5 times on average after fertilization, although the number of cell division rounds differs among tissues. This article discusses the possible mechanisms that control the timing of the final cell division: these include (i) the analog and digital clock models, in which the elapsed time and number of cell division cycles are measured after fertilization, respectively, and (ii) the analog and digital timer models, in which the elapsed time and number of cell divisions are measured after cell fate specification, respectively. In notochord and muscle cells of ascidian embryos, cell division termination involves cell fate specification by inductive FGF signaling, followed by activation of the tissue-specific key transcription factors, Brachyury and Tbx6, which then induce gene expression of an effector, Cdk inhibitor. The present evidence suggests that these timing mechanisms comprise two functional phases: (i) an analog timer, which is mediated by accumulation of Cdk inhibitor protein after cell fate specification, and (ii) a timekeeper, which confines the timing, when the above timer is triggered by Brachyury and Tbx6, to the appropriate stage.


Constitutive expression of thrombospondin 1 in MC3T3-E1 osteoblastic cells inhibits mineralization.
Thrombospondin 1 (TSP1) is a multifunctional extracellular glycoprotein present mainly in the fetal and adult skeleton. Although an inhibitory effect of TSP1 against pathological mineralization in cultured vascular pericytes has been shown, its involvement in physiological mineralization by osteoblasts is still unknown. To determine the role of TSP1 in biomineralization, mouse osteoblastic MC3T3-E1 cells were cultured in the presence of antisense phosphorothioate oligodeoxynucleotides complementary to the TSP1 sequence. The 18- and 24-mer antisense oligonucleotides caused concentration-dependent increases in the number of mineralized nodules, acid-soluble calcium deposition in the cell/matrix layer, and alkaline phosphatase activity within 9 days, without affecting cell proliferation. The corresponding sense or scrambled oligonucleotides did not affect these parameters. In the antisense oligonucleotide-treated MC3T3-E1 cells, thickened extracellular matrix, well-developed cell processes, increased intracellular organelles, and collagen fibril bundles were observed. On the other hand, the addition of TSP1 to the culture decreased the production of a mineralized matrix by MC3T3-E1 cells. Furthermore, MC3T3-E1 clones overexpressing mouse TSP1 were established and assayed for TSP1 protein and their capacity to mineralize. TSP1 dose-dependently inhibited mineralization by these cells both in vitro and in vivo. These results indicate that TSP1 functions as an inhibitory regulator of bone mineralization and matrix production by osteoblasts to sustain bone homeostasis.


Honokiol suppresses metastasis of renal cell carcinoma by targeting KISS1/KISS1R signaling.
Renal cell carcinoma (RCC) is a common urological cancer worldwide and is known to have a high risk of metastasis, which is considered responsible for more than 90% of cancer associated deaths. Honokiol is a small-molecule biphenol isolated from Magnolia spp. bark and has been shown to be a potential anticancer agent involved in multiple facets of signal transduction. In this study, we demonstrated that honokiol inhibited the invasion and colony formation of highly metastatic RCC cell line 786-0 in a dose-dependent manner. DNA-microarray data showed the significant upregulation of metastasis-suppressor gene KISS1 and its receptor, KISS1R. The upregulation was confirmed by qRT-PCR analysis. Overexpression of KISS1 and KISS1R was detected by western blotting at the translation level as well. Of note, the decreased invasive and colonized capacities were reversed by KISS1 knockdown. Taken together, the results first indicate that activation of KISS1/KISS1R signaling by honokiol suppresses multistep process of metastasis, including invasion and colony formation, in RCC cells 786-0. Honokiol may be considered as a natural agent against RCC metastasis.


Macrophage infiltration in 3D cancer spheroids to recapitulate the TME and unveil interactions within cancer cells and macrophages to modulate chemotherapeutic drug efficacy.
Recapitulating the tumor microenvironment (TME) in vitro remains a major hurdle in cancer research. In recent years, there have been significant strides in this area, particularly with the emergence of 3D spheroids as a model system for drug screening and therapeutics development for solid tumors. However, incorporating macrophages into these spheroid cultures poses specific challenges due to the intricate interactions between macrophages and cancer cells. To address this issue, in this study, we established a reproducible healthy multicellular 3D spheroid culture with macrophage infiltrates in order to mimic the TME and modulate the drug's efficacy on cancer cells in the presence of macrophages. A 3D spheroid was established using the human cancer cell line CAL33 and THP1 cell derived M0 macrophages were used as a source of macrophages. Cellular parameters including tumour metabolism, health, and mitochondrial mass were analysed in order to establish ideal conditions. To modulate the interaction of cancer cells with macrophage the ROS, NO, and H2O2 levels, in addition to M1 and M2 macrophage phenotypic markers, were analyzed. To understand the crosstalk between cancer cells and macrophages for ECM degradation, HSP70, HIF1α and cysteine proteases were examined in spheroids using western blotting and qPCR. The spheroids with macrophage infiltrates exhibited key features of solid tumors, including cellular heterogeneity, metabolic changes, nutrient gradients, ROS emission, and the interplay between HIF1α and HSP70 for upregulation of ECM degradading enzymes. Our results demonstrate that tumor cells exhibit a metabolic shift in the presence of macrophages. Additionally, we have observed a shift in the polarity of M0 macrophages towards tumor-associated macrophages (TAMs) in response to cancer cells in spheroids. Results also demonstrate the involvement of macrophages in regulating HIF-1α, HSP70, and ECM degradation cysteine proteases enzymes. This study has significant implications for cancer therapy as it sheds light on the intricate interaction between tumor cells and their surrounding macrophages. Additionally, our 3D spheroid model can aid in drug screening and enhance the predictive accuracy of preclinical studies. The strength of our study lies in the comprehensive characterization of the multicellular 3D spheroid model, which closely mimics the TME.


Synthesis and evaluation of 2-ethynyl-adenosine-5'-triphosphate as a chemical reporter for protein AMPylation.
Protein AMPylation is a posttranslational modification (PTM) defined as the transfer of an adenosine monophosphate (AMP) from adenosine triphosphate (ATP) to a hydroxyl side-chain of a protein substrate. One recently reported AMPylator enzyme, Vibrio outer protein S (VopS), plays a role in pathogenesis by AMPylation of Rho GTPases, which disrupts crucial signaling pathways, leading to eventual cell death. Given the resurgent interest in this modification, there is a critical need for chemical tools that better facilitate the study of AMPylation and the enzymes responsible for this modification. Herein we report the synthesis of 2-ethynyl-adenosine-5'-triphosphate () and its utilization as a non-radioactive chemical reporter for protein AMPylation.


Strategies for evaluation of suitable donors: Italian experience.
Italy was lacking standardised procedures for donor safety evaluation. We developed practice guidelines, while a panel of experts coordinated by the National Transplant Centre, is available 24 hours a day to support decisions in difficult cases. The guidelines identify five levels of risk and give recommendations for the utilization of donors with HBV and HCV infections as well as for malignancies with negligible or very low risk of transmission. In conclusion we aim to standardize the process of donor evaluation across Italy, to increase the pool of utilised donors and to reduce the risk of communicable disease transmission.


Modelling the bronchial barrier in pulmonary drug delivery: A human bronchial epithelial cell line supplemented with human tracheal mucus.
The airway epithelium together with the mucus layer coating it forms a protective system that efficiently filters and removes potentially harmful particles contained in inhaled air. The same mechanism, however, serves to entrap particulate drug carriers, precluding their interaction with their target. The mucus barrier is often neglected in in vitro testing setups employed for the assessment of pulmonary drug delivery strategies. Therefore, our aim was to more accurately model the bronchial barrier, by developing an in vitro system comprising a tight epithelial cell layer which may be optionally supplemented with a layer of human tracheal mucus. To form the epithelium in vitro, we used the cystic fibrosis cell line CFBE41o-, which can be grown as monolayers on Transwell® supports, expressing tight junctions as well as relevant transport proteins. In contrast to the cell line Calu-3, however, CFBE41o- does not produce mucus. Therefore, native human mucus, obtained from tracheal tubes of patients undergoing elective surgery, was used as a supplement. The compatibility of CFBE41o- cells with the human mucus was addressed with the MTT assay, and confirmed by fluorescein diacetate/propidium iodide live/dead staining. Moreover, the CFBE41o- cells retained their epithelial barrier properties after being supplemented with mucus, as evidenced by the high trans-epithelial electrical resistance values (∼1000Ωcm2) together with a continued low level of paracellular transport of sodium fluorescein. Fluorescently-labeled chitosan-coated PLGA nanoparticles (NP, ∼168nm) were used as a model drug delivery system to evaluate the suitability of this in vitro model for studying mucus permeation and cell uptake. Comparing CFBE41o- cell monolayers with and without mucus, resp., showed that the NP uptake was dramatically reduced in the presence of mucus. This model may therefore be used as a tool to study potential mucus interactions of aerosolized drugs, and more specifically NP-based drug delivery systems designed to exert their effect in the bronchial region.


CT signs of urethral injury.
Computed tomography (CT) is the accepted frontline imaging modality for blunt abdominopelvic trauma. However, urethral injuries are traditionally diagnosed with retrograde urethrography. The CT appearances of urethral injuries and the signs associated with posterior urethral injuries are not well described in the literature. CT scans of patients with pelvic fractures and urethrographically proved posterior urethral injuries were evaluated. CT scans of patients with similar pelvic fractures who did not have urethral injuries were also evaluated. The CT findings of elevation of the prostatic apex, extravasation of urinary tract contrast material above the urogenital diaphragm (UGD), and extravasation of urinary tract contrast material below the UGD were specific for type I, II, and III urethral injuries, respectively. If extraperitoneal bladder rupture is present along with periurethral extravasation of contrast material, the possibility of type IV and IVA urethral injuries should be considered. In addition, the CT findings of distortion or obscuration of the UGD fat plane, hematoma of the ischiocavernosus muscle, distortion or obscuration of the prostatic contour, distortion or obscuration of the bulbocavernosus muscle, and hematoma of the obturator internus muscle were more common in patients with pelvic fractures and associated urethral injuries than in patients with uncomplicated pelvic fractures.


Increased Hydrophobic Block Length of PTDMs Promotes Protein Internalization.
The plasma membrane is a major obstacle in the development and use of biomacromolecules for intracellular therapeutic applications. Protein transduction domains (PTDs) have been used to overcome this barrier, but often require covalent conjugation to their cargo and can be time consuming to synthesize. Synthetic monomers can be designed to mimic the amino acid moieties in PTDs, and their resulting polymers provide a well-controlled platform to vary molecular composition for structure-activity relationship studies. In this paper, a series of polyoxanorbornene-based synthetic mimics, inspired by PTDs, with varying cationic and hydrophobic densities, and the nature of the hydrophobic chain and degree of polymerizations were investigated in vitro to determine their ability to non-covalently transport enhanced green fluorescent protein into HeLa cells, Jurkat T cells, and hTERT mesenchymal stem cells. Polymers with high charge density lead to efficient protein delivery. Similarly, the polymers with the highest hydrophobic content and density proved to be the most efficient at internalization. The observed improvements with increased hydrophobic length and content were consistent across all three cell types, suggesting that these architectural relationships are not cell type specific. However, Jurkat T cells showed distinct variation in uptake between polymers than with the other two cell types. These results provide important design parameters for more effective delivery of biomacromolecules for intracellular delivery applications.


Neural activation during experimental allodynia: a functional magnetic resonance imaging study.
Abstract Pain induced by gentle stroking, i.e. dynamic-mechanical allodynia, is one of the most distressing symptoms of neuropathic pain. The underlying neuronal pathways are still a matter of debate. Here, we investigated the cortical activations associated with dynamic-mechanical allodynia in an experimental human pain model by functional magnetic resonance imaging (fMRI). Large and stable areas of brush-evoked allodynia were induced in 11 healthy subjects by topical capsaicin (2.5%, 30 min) application following local heating (45 degrees C for 5 min), thus combining both physical and chemical sensitization. During the fMRI experiments, allodynia was rekindled by local heat application (40 degrees C for 5 min) immediately before the allodynia testing. Brushing the untreated forearm (control condition) led to activations of the contralateral primary somatosensory cortex (S1), contralateral parietal association cortex (PA), bilateral secondary somatosensory cortices (S2) and insula (contralateral). Brushing the allodynic skin was painful and the cortical responses were partially overlapping with those induced by the nonpainful brush stimulation. Additionally, the contralateral inferior frontal cortex (IFC) and the ipsilateral insula were activated. Direct comparison between nonpainful brushing and brush-evoked allodynia revealed significant increases in blood oxygenation level-dependent (BOLD) signals in contralateral S1, PA, IFC and bilateral S2/insula during allodynia. This study highlights the importance of a cortical network comprising S1, PA, S2/insula and IFC in the processing of dynamic-mechanical allodynia in the human brain. Furthermore, it demonstrates that the combined heat/capsaicin model can be used successfully in the exploration of brain processes underlying stimulus-evoked pain.


Drug reaction with eosinophilia and systemic symptoms without skin rash.
Drug reaction with eosinophilia and systemic symptoms (DRESS) or drug hypersensitivity syndrome is considered as a severe cutaneous adverse drug reaction which is most commonly precipitated by aromatic anticonvulsants, lamotrigine, dapsone, allopurinol, minocycline, and salazopyrin. Its clinical manifestations are often variable. On rare occasions, it can present with only systemic involvement without any cutaneous features. A complete drug history is of paramount importance in making an early diagnosis. We report the case of a male patient who presented with fever, lymphadenopathy, hepatosplenomegaly, and hepatitis, 2 weeks after starting salazopyrin. The presence of atypical lymphocytes in the peripheral smear was indicative of a viral infection or a hematological dyscrasia. Bone marrow examination revealed a normocellular marrow with an increase in eosinophil precursors. Investigations for the common causes for fever and hepatitis were negative. The presence of eosinophilia, the temporal relationship of the symptoms with the initiation of treatment with salazopyrin, and the marked improvement on withdrawal of the drug along with the administration of systemic corticosteroids, were features consistent with the diagnosis of DRESS. With the incidence of this condition showing a rising trend, it is important for the clinician to be aware of its variable manifestations, as a delay in diagnosis and treatment can be fatal.


Haptoglobin interacts with apolipoprotein E and beta-amyloid and influences their crosstalk.
Beta-amyloid accumulation in brain is a driving force for Alzheimer's disease pathogenesis. Apolipoprotein E (ApoE) represents a critical player in beta-amyloid homeostasis, but its role in disease progression is controversial. We previously reported that the acute-phase protein haptoglobin binds ApoE and impairs its function in cholesterol homeostasis. The major aims of this study were to characterize the binding of haptoglobin to beta-amyloid, and to evaluate whether haptoglobin affects ApoE binding to beta-amyloid. Haptoglobin is here reported to form a complex with beta-amyloid as shown by immunoblotting experiments with purified proteins, or by its immunoprecipitation in brain tissues from patients with Alzheimer's disease. The interaction between ApoE and beta-amyloid was previously shown to be crucial for limiting beta-amyloid neurotoxicity and for promoting its clearance. We demonstrate that haptoglobin, rather than impairing ApoE binding to beta-amyloid, promotes to a different extent the formation of the complex between beta-amyloid and ApoE2 or ApoE3 or ApoE4. Our data suggest that haptoglobin and ApoE functions in brain should be evaluated taking into account their mutual interaction with beta-amyloid. Hence, the risk of developing Alzheimer's disease might not only be linked to the different ApoE isoforms, but also rely on the level of critical ligands, such as haptoglobin.


Genetically encoded impairment of neuronal KCC2 cotransporter function in human idiopathic generalized epilepsy.
The KCC2 cotransporter establishes the low neuronal Cl(-) levels required for GABAA and glycine (Gly) receptor-mediated inhibition, and KCC2 deficiency in model organisms results in network hyperexcitability. However, no mutations in KCC2 have been documented in human disease. Here, we report two non-synonymous functional variants in human KCC2, R952H and R1049C, exhibiting clear statistical association with idiopathic generalized epilepsy (IGE). These variants reside in conserved residues in the KCC2 cytoplasmic C-terminus, exhibit significantly impaired Cl(-)-extrusion capacities resulting in less hyperpolarized Gly equilibrium potentials (EG ly), and impair KCC2 stimulatory phosphorylation at serine 940, a key regulatory site. These data describe a novel KCC2 variant significantly associated with a human disease and suggest genetically encoded impairment of KCC2 functional regulation may be a risk factor for the development of human IGE.


Studies of a controlled-release microcapsule formulation of an LH-RH agonist (D-Trp-6-LH-RH) in the rhesus monkey menstrual cycle.
It has been shown that continuous administration of LH-RH agonists inhibits ovulation in many species, including human and nonhuman primates. However, the need of daily injections or intranasal application of the LH-RH agonists makes this contraceptive approach impractical. In addition, due to the repeated daily administration of the LH-RH analogues and their bioavailability, serum levels of gonadotropins and estradiol present wide variations and are often associated with irregular bleeding and/or endometrial hyperplasia. This experiment was designed to study the effects on ovulation and the hormonal profile during the rhesus monkey menstrual cycle of a system that continuously delivers a potent agonist of LH-RH (D-Trp-6-LH-RH encapsulated in poly [DL-lactide-co-glycolide]). Subjects were divided into two groups: On day 1 of the cycle, monkeys received a single subcutaneous injection of microcapsules containing D-Trp-6-LH-RH at a release rate of 13.6 micrograms/day (Group 1) or placebo microcapsules (Group 2). Ovulation dates were significantly delayed in animals from Group 1 (50 +/- 3 days) as compared to those in group 2 (20 +/- 2 days). Luteal phases following ovulation were normal in animals of both groups, as determined by length and serum progesterone levels. No significant differences on baseline levels of FSH and LH were observed between the two groups of animals. Post-treatment cycles were ovulatory and presented normal luteal phases and hormone concentrations as compared to the non-treated animals of our colony. The present data show for the first time that a controlled-release microcapsule formulation that delivers an agonist of LH-RH can effectively suppress ovulation when injected in a single dose in non-human primates.


Physical therapy management of a patient with Eagle syndrome.
The purpose of this care report was to describe the physical therapy management of a unique case involving a 50-year-old white female with complaints pertaining to the craniofacial, cervical, thoracic, and brachial regions with known Eagle syndrome as a complicating factor. Intervention consisted of manual therapy and therapeutic exercises. The Craniocervical Flexion Test (CCFT), Neck Disability Index (NDI), and subjective complaints were used as outcome measures. The patient was tested at initial visit, discharge, and at a 3-month follow-up. At discharge the patient reported complete cessation of craniofacial symptoms, her CCFT score had improved by 4 mmHg, and the NDI score had improved by 10%. At the 3-month follow-up, the patient's CCFT had improved an additional 4 mmHg and the NDI score had declined by 4%. The cumulative outcomes were an 8 mmHg improvement in the CCFT and a 6% increase on the NDI. Although subjective complaints and CCFT scores improved, clinically significant improvement with the NDI was not maintained. Limitations of this case study include the substitution of standardized equipment for the CCFT and the presence of numerous comorbidities that possibly influenced results. Further research is necessary.


Three patients who spontaneously developed persistent hypothyroidism during or following treatment with antithyroid drugs for Graves' hyperthyroidism.
Three patients with Graves' disease who spontaneously developed hypothyroidism after treatment with antithyroid drugs are described herein. Patient 1 developed a painful tender thyroid enlargement with a fever and accelerated erythrocyte sedimentation rate when she was receiving maintenance therapy with methimazole, and she progressed to persistent hypothyroidism with increased titers of antithyroglobulin and antimicrosomal antibodies and marked reduction of goiter size within the subsequent 2 months. Thyroid-stimulating hormone-binding inhibitory immunoglobulins (TBIIs) and thyroid stimulation-blocking antibody (TSBAb) were absent when she was hypothyroid. Hypothyroidism probably resulted from autoimmune thyroid destruction due to subacute aggravation of Hashimoto's thyroiditis. During the clinical course of patient 2, accelerated erythrocyte sedimentation rate and later transient increases of antimicrosomal and antithyroglobulin antibody titers were observed repeatedly (four times), and she finally fell into overt hypothyroidism. She also had negative results of tests for TBII and TSBAb. Her hypothyroidism appeared to result from repeated thyroid destruction due to aggravation of Hashimoto's thyroiditis. Patient 3 fell into hypothyroidism when receiving a small dosage of methimazole. The TBII and TSBAb were strongly active when she developed hypothyroidism, which thus seemed to be due to blocking antibody. Patients with Graves' hyperthyroidism may eventually progress to hypothyroidism later by several different mechanisms. Severe and sudden or slowly repeated thyroid destruction due to aggravation of Hashimoto's thyroiditis is one mechanism. Another may be the appearance of a blocking antibody to the TSH receptor.


Clinical analysis of esophageal cancer patients with a history of metachronous primary cancer.
Esophageal cancer (EC) is often associated with multiple primary cancers, and the frequency of EC in patients with a history of metachronous primary cancers (HMPC) was reported as 7.8%. The clinical characteristics of these cases have not yet been elucidated in detail. Therefore, we analyzed 36 EC patients with HMPC among 370 cases that underwent curative resection in our hospital between 1996 and 2013. The most frequent HMPC was gastric cancer (36.1%). The group with HMPC was significantly older than the group without HMPC, whereas no significant differences were observed in other background factors. The stomach was used for reconstruction more frequently reconstructed in the group without HMPC (p<.05). No significant differences were observed in surgical complications between the 2 groups (p=0.283). The 5-year overall survival rates were similar between the groups. The safety of surgery for and prognosis of EC patients with HMPC were similar to those of EC patients without HMPC; therefore, curative surgery should be considered for EC patients with HMPC.


Generation of murine xenograft models of brain tumors from primary human tissue for in vivo analysis of the brain tumor-initiating cell.
The generation of xenograft models, which support the growth of human tissue in animals, forms an important part of a researcher's tool kit and enhances the ability to understand the initiation and development of cancer in vivo. Especially in the context of the brain tumor-initiating cell (BTIC), a xenograft model allows for careful characterization of BTIC roles in tumor initiation, growth, and relapse. Here, we detail a set of procedures which describe the isolation, enrichment, and intracranial injection of human BTICs from patient samples to generate xenograft models of a human brain tumor.


Impacts of parent-implemented early-literacy intervention for Spanish-speaking children with language impairment.
Children with language impairment (LI) often have lags in development of print knowledge, an important early-literacy skill. This study explores impacts of a print-focused intervention for Spanish-speaking children with LI in Southeastern Mexico. Aims were twofold. First, we sought to describe the print knowledge (print-concept knowledge, alphabet knowledge) of Spanish-speaking children with LI. Second, we determined the extent to which print-referencing intervention delivered by children's parents could improve print knowledge. Using a pre-test-post-test delayed treatment research design, 13 parent-child dyads were assigned to an intervention (n = 8) versus control (n = 5) condition. Children were drawn from a speech-language clinic and all were receiving services for LI. Caregivers in the intervention group implemented an 8-week home-reading programme following a systematic scope and sequence for improving children's print knowledge. Children showed individual differences in their print knowledge based on three baseline measures examining print-concept knowledge, alphabet knowledge and letter-sound knowledge. Those whose caregivers implemented the 8-week programme showed statistically and practically significant gains on two of the three measures over the intervention period. The results presented here may stimulate future research on the print knowledge of Spanish-speaking children with LI. Sources of individual differences are important to determine. Caregivers may use the intervention presented here as a potential avenue for improving children's print knowledge.


[Functional concept of the initiation of blood coagulation by cell membranes].
According to the conception proposed, blood coagulation is initiated by Ca(2+)-induced rearrangement of the bilayer structure of native cell membranes into heterophasic. The thrombogenic action is realized through Ca2+ entering the cell cytoplasm. This leads to phosphatidyl serine translocation into the surface monolayer of the membrane, its clustering and the arising of phosphatidyl ethanolamine mesophases. Vitamin K-dependent coagulation factors are bound at the boundaries of clusters and mesophases of these phospholipids through Ca2+ ions. They form enzyme complexes functioning due to the matrix structure of phospholipid surface as units of thrombin generation.


National surveillance data on the epidemiology of cholera in Kenya, 1997-2010.
Kenya has experienced multiple cholera outbreaks since 1971. Cholera remains an issue of major public health importance and one of the 35 priority diseases under Kenya's updated Integrated Disease Surveillance and Response strategy. We reviewed the cholera surveillance data reported to the World Health Organization and the Kenya Ministry of Public Health and Sanitation from 1997 through 2010 to determine trends in cholera disease for the 14-year period. A total of 68 522 clinically suspected cases of cholera and 2641 deaths were reported (overall case-fatality rate [CFR], 3.9%), affecting all regions of the country. Kenya's largest outbreak occurred during 1997-1999, resulting in 26 901 cases and 1362 deaths (CFR, 5.1%). Following a decline in disease occurrence, the country experienced a resurgence of epidemic cholera during 2007-2009 (16 616 cases and 454 deaths; CFR, 2.7%), which declined rapidly to 0 cases. Cases were reported through July 2010, with no cases reported during the second half of the year. About 42% of cases occurred in children aged <15 years. Vibrio cholerae O1, serotype Inaba, was the predominant strain recorded from 2007 through 2010, although serotype Ogawa was also isolated. Recurrent outbreaks have most frequently affected Nairobi, Nyanza, and Coast provinces, as well as remote arid and semiarid regions and refugee camps. Kenya has experienced substantial amounts of reported cases of cholera during the past 14 years. Recent decreases in cholera case counts may reflect cholera control measures put in place by the National Ministry of Health; confirmation of this theory will require ongoing surveillance.


Cardiovascular protective effects of n-3 polyunsaturated fatty acids with special emphasis on docosahexaenoic acid.
It is widely accepted that n-3 polyunsaturated fatty acids (PUFAs) rich in fish oils protect against several types of cardiovascular diseases such as myocardial infarction, arrhythmia, atherosclerosis, or hypertension. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) may be the active biological components of these effects. Although the precise cellular and molecular mechanisms underlying the beneficial effects are still uncertain, the protective effects of n-3 PUFAs are attributable to their direct effects on vascular smooth muscle cell (VSMC) functions. These n-3 PUFAs activate K(+)(ATP) channels and inhibit certain types of Ca(2+) channels, probably via at least 2 distinct mechanisms. N-3 PUFAs favorably alter the eicosanoid profile and regulate cytokine-induced expression of inducible nitric oxide synthase and cyclooxygenase-2 via mechanisms involving modulation of signaling transduction events. N-3 PUFAs also modulate VSMC proliferation, migration, and apoptosis. These recent data suggest that modulation of these VSMC functions contribute to the beneficial effects of n-3 PUFAs on various cardiovascular disorders. Furthermore, recent studies strongly suggest that DHA has more potent and beneficial effects than EPA. However, many questions about the cellular and molecular mechanisms still remain to be answered.


In vitro and in vivo models of celiac disease.
Background : Celiac disease is an autoimmune intestinal disease that develops in genetically susceptible individuals upon the ingestion of gluten. In vitro and in vivo models have addressed the roles of the following five elements in the pathogenesis of celiac disease: i) A tight association with human leukocyte antigen (HLA)-DQ2 and HLA-DQ8. ii) A strong B-cell response to the autoantigen tissue transglutaminase that preferentially reacts with certain gluten peptides and that is almost exclusively IgA. iii) An adaptive inflammatory immune response to gluten peptides that are antigenically presented on HLA-DQ2 or HLA-DQ8. iv) An improved antigenic presentation of most gluten peptides and a stronger T-cell response after their deamidation by tissue transglutaminase; and v) An increased risk for autoimmune diseases associated with celiac disease, such as dermatitis herpetiformis and type 1 diabetes. To summarize the different in vitro and in vivo models of celiac disease, their contributions to our understanding of the pathogenesis of celiac disease and the alternative therapies that have been tested using these models. These models, especially the animal models, are continuously being developed. They have contributed greatly to the development of alternative therapies for celiac disease and are being used increasingly to test their effectiveness and potential.


The absorption, distribution and excretion of anabolic agents.
The metabolic fate of the anabolic agents, diethylstilbestrol, hexestrol, trenbolone acetate, zeranol and the endogenous steroids are discussed under the headings absorption, distribution and excretion. There is an optimum concentration of anabolic agent in the systemic circulation that results in a maximum increase in growth rate of farm animals. This optimum blood concentration should be maintained over a long period. However, there is rapid metabolism and excretion of anabolic agents with short half-lives in blood, and metabolic clearance rate equals entry rate. The rate of absorption of the agent, which is determined by formulation and site of administration, is most important and is best achieved by the use of slow release implants. The pattern of exponential absorption from compressed pellets of single anabolic agents is not ideal, and a more constant payout of drug, in particular estradiol-17 beta, is best achieved in combined preparations of agents or from silicone rubber implants impregnated with the agent. The high metabolic clearance rate and rapid excretion of anabolic substances influences the distribution of residues. Outside the site of administration, less than 1% of the administered dose is present in the animal. The lowest concentrations of residues are found in muscle and fat, higher concentrations are present in liver and kidney and the highest concentrations are in the bile, urine and feces.


Vascular anomalies of the hand and wrist.
Vascular malformations of the hand and wrist are uncommon. They develop from aberrations in angiogenic signaling during vascular development. Unlike hemangiomas, which are characterized by biphasic growth and slow spontaneous involution, vascular malformations continue to grow proportionally with the child. Management is dictated by classification of the vascular malformations, which is based on flow characteristics (ie, low, high) and predominant cell type (ie, venous, lymphatic, capillary, combined, arteriovenous). Initial management is conservative, with the goal of providing relief from pain and swelling. Sclerotherapy, laser treatment, and arterial embolization may be beneficial in well-selected patients. Surgery is indicated in cases of persistent pain and uncontrolled limb swelling leading to functional impairment and/or neurologic compression. The goals of surgery are to excise as much of the lesion as possible while avoiding injury to adjacent nerves, minimizing blood loss, and preventing distal limb ischemia. This mandates careful preoperative planning and meticulous technique. Adjuvant treatments may be warranted, as in the case of preoperative embolization in patients with high-flow lesions.


Public health infrastructure: creating a solid foundation.
The nation's public health system remains in disarray, despite the 1988 warning by the Institute of Medicine in its landmark report, "The Future of Public Health." Recent concerns about antibiotic resistance, inadequate disease surveillance capabilities, bioterrorism, and an increasing need for training the public health workforce have brought this problem into the forefront of congressional concern. Recent legislation aimed at addressing this problem was passed last Fall and signed into law. This program will take a significant step towards ensuring a solid public health system when it is fully implemented.


Retrospective analysis of 23 cases with peripheral T-cell lymphoma, unspecified: clinical characteristics and outcome.
Peripheral T-cell lymphomas (PTCL) are a heterogeneous group of post-thymic malignancies relatively uncommon in the Western world and their prognosis and therapeutic approach are still not well defined. The aim of this study was to retrospectively analyze the clinical, hematological and histological features at diagnosis, the relevance of the International Prognostic Index and the outcome of a group of 23 patients affected by peripheral T-cell lymphoma, unspecified (PTCL-U), according to the Revised European-American Classification of Lymphoid Neoplasms (REAL), observed between September 1985 and April 1995 at our Institution. Patients were separated into different prognostic groups according to Ann Arbor stage, cell size and International Prognostic Index. All patients had been treated with multiagent combination chemotherapy, mainly CHOP (9 cases) and F-MACHOP (9 cases), and were evaluable for response. The treatment was intensified with allogeneic bone marrow transplantation (BMT) in 1 patient and with autologous BMT in 4 patients. Median age was 55 (range 18-77) years and 70% of the patients were males. Four patients were in stage II (17%), 5 in stage III (22%) and 14 in stage IV (61%). Patient risk was classified according to the International Prognostic Index as follows: 8 cases (35%) low risk, 2 cases (9%) low-intermediate, 8 cases (35%) high-intermediate, 5 cases (21%) high. Median follow-up time was 20 months (range 2-132). Median progression-free survival (PFS) and overall survival (OS) for all the patients studied were 10 and 34 months, respectively. Stage IV was associated with a poorer response rate and a shorter PFS (median 6 months) and OS (median 32 months). No statistical correlation was found between cell size and overall response (complete + partial remission), PFS (p = 0.38) or OS (p = 0.59), although a better trend was observed for the large cell group. A less favorable outcome was observed in patients in the high-intermediate + high risk groups, where median PFS and OS were 7 and 24 months, respectively, than in patients in the low + low-intermediate risk groups. No difference in response or outcome was detected between patients treated with the CHOP and the F-MACHOP regimens, while all 5 patients given high-dose chemotherapy and BMT are alive and in CR. Our experience shows that PTCL-U are rare lymphomas frequently having an aggressive presentation. The response to conventional polychemotherapeutic regimens like CHOP or F-MACHOP is generally poor, especially in those cases with advanced stage and a high-intermediate or high International Prognostic Index. The observation that all five patients who were treated with bone marrow transplantation are alive and in complete remission suggests using this strategy, particularly in young patients with a poor International Prognostic Index.


Industrial Ziegler-type hydrogenation catalysts made from Co(neodecanoate)2 or Ni(2-ethylhexanoate)2 and AlEt3: evidence for nanoclusters and sub-nanocluster or larger Ziegler-nanocluster based catalysis.
Ziegler-type hydrogenation catalysts are important for industrial processes, namely, the large-scale selective hydrogenation of styrenic block copolymers. Ziegler-type hydrogenation catalysts are composed of a group 8-10 transition metal precatalyst plus an alkylaluminum cocatalyst (and they are not the same as Ziegler-Natta polymerization catalysts). However, for ∼50 years two unsettled issues central to Ziegler-type hydrogenation catalysis are the nature of the metal species present after catalyst synthesis, and whether the species primarily responsible for catalytic hydrogenation activity are homogeneous (e.g., monometallic complexes) or heterogeneous (e.g., Ziegler nanoclusters defined as metal nanoclusters made from combination of Ziegler-type hydrogenation catalyst precursors). A critical review of the existing literature (Alley et al. J. Mol. Catal. A: Chem. 2010, 315, 1-27) and a recently published study using an Ir model system (Alley et al. Inorg. Chem. 2010, 49, 8131-8147) help to guide the present investigation of Ziegler-type hydrogenation catalysts made from the industrially favored precursors Co(neodecanoate)(2) or Ni(2-ethylhexanoate)(2), plus AlEt(3). The approach and methods used herein parallel those used in the study of the Ir model system. Specifically, a combination of Z-contrast scanning transmission electron microscopy (STEM), matrix assisted laser desorption ionization mass spectrometry (MALDI MS), and X-ray absorption fine structure (XAFS) spectroscopy are used to characterize the transition metal species both before and after hydrogenation. Kinetic studies including Hg(0) poisoning experiments are utilized to test which species are the most active catalysts. The main findings are that, both before and after catalytic cyclohexene hydrogenation, the species present comprise a broad distribution of metal cluster sizes from subnanometer to nanometer scale particles, with estimated mean cluster diameters of about 1 nm for both Co and Ni. The XAFS results also imply that the catalyst solutions are a mixture of the metal clusters described above, plus unreduced metal ions. The kinetics-based Hg(0) poisoning evidence suggests that Co and Ni Ziegler nanoclusters (i.e., M(≥4)) are the most active Ziegler-type hydrogenation catalysts in these industrial systems. Overall, the novelty and primary conclusions of this study are as follows: (i) this study examines Co- and Ni-based catalysts made from the actual industrial precursor materials, catalysts that are notoriously problematic regarding their characterization; (ii) the Z-contrast STEM results reported herein represent, to our knowledge, the best microscopic analysis of the industrial Co and Ni Ziegler-type hydrogenation catalysts; (iii) this study is the first explicit application of an established method, using multiple analytical methods and kinetics-based studies, for distinguishing homogeneous from heterogeneous catalysis in these Ziegler-type systems; and (iv) this study parallels the successful study of an Ir model Ziegler catalyst system, thereby benefiting from a comparison to those previously unavailable findings, although the greater M-M bond energy, and tendency to agglomerate, of Ir versus Ni or Co are important differences to be noted. Overall, the main result of this work is that it provides the leading hypothesis going forward to try to refute in future work, namely, that sub, M(≥4) to larger, M(n) Ziegler nanoclusters are the dominant, industrial, Co- and Ni- plus AlR(3) catalysts in Ziegler-type hydrogenation systems.


A questionnaire survey on attitudes and understanding towards mental disorders.
OBJECTIVES. To obtain information about basic knowledge towards mental disorders and to evaluate public attitudes towards mental disorders in the Hong Kong Chinese population. METHODS. Questionnaires which collected basic demographic information, opinions about potential stigmas and myths, and knowledge on case vignettes depicting fictional characters with symptoms of mental illness were delivered to subjects in a secondary school, 2 homes for the elderly, a private housing estate, and a public housing estate in Hong Kong. RESULTS. Completed questionnaires were collected from 1035 subjects. In general, the participants' acceptance of mental illness was good. Regular contacts with such patients were associated with better knowledge (t = -2.71, p < 0.01) and better acceptance (t = 2.77, p < 0.01) of mental illness. Younger participants aged 15 to 19 years had a lower level of knowledge about mental health problems compared with other age-groups (p < 0.001). CONCLUSIONS. Personal contact with people with mental illness may help to improve knowledge and acceptance. Younger people in secondary school should be the target and prioritised group for mental health education. Apart from the delivery of mental health knowledge, strategies to increase social contact of the public with people having mental illness could be considered in the design and implementation of anti-stigma programmes.


Multiplexing detection of IgG against Plasmodium falciparum pregnancy-specific antigens.
Pregnant women exposed to Plasmodium falciparum generate antibodies against VAR2CSA, the parasite protein that mediates adhesion of infected erythrocytes to the placenta. There is a need of high-throughput tools to determine the fine specificity of these antibodies that can be used to identify immune correlates of protection and exposure. Here we aimed at developing a multiplex-immunoassay to detect antibodies against VAR2CSA antigens. We constructed two multiplex-bead arrays, one composed of 3 VAR2CSA recombinant-domains (DBL3X, DBL5Ɛ and DBL6Ɛ) and another composed of 46 new peptides covering VAR2CSA conserved and semi-conserved regions. IgG reactivity was similar in multiplexed and singleplexed determinations (Pearson correlation, protein array: R2 = 0.99 and peptide array: R2 = 0.87). IgG recognition of 25 out of 46 peptides and all recombinant-domains was higher in pregnant Mozambican women (n = 106) than in Mozambican men (n = 102) and Spanish individuals (n = 101; p<0.05). Agreement of IgG levels detected in cryopreserved plasma and in elutions from dried blood spots was good after exclusion of inappropriate filter papers. Under heterogeneous levels of exposure to malaria, similar seropositivity cutoffs were obtained using finite mixture models applied to antibodies measured on pregnant Mozambican women and average of antibodies measured on pregnant Spanish women never exposed to malaria. The application of the multiplex-bead array developed here, allowed the assessment of higher IgG levels and seroprevalences against VAR2CSA-derived antigens in women pregnant during 2003-2005 than during 2010-2012, in accordance with the levels of malaria transmission reported for these years in Mozambique. The multiplex bead-based immunoassay to detect antibodies against selected 25 VAR2CSA new-peptides and recombinant-domains was successfully implemented. Analysis of field samples showed that responses were specific among pregnant women and dependent on the level of exposure to malaria. This platform provides a high-throughput approach to investigating correlates of protection and identifying serological markers of exposure for malaria in pregnancy.


Approaches to improving breast cancer diagnosis using a high resolution, breast specific gamma camera.
Mammography remains the gold standard in breast cancer detection, although there remains a challenge for improvement in sensitivity of breast cancer detection and diagnosis. Although mammography is the most frequently utilized examination to screen for breast cancer, which has resulted in a reduction of breast cancer mortality, still some cancers are unable to be visualized on mammographic images. Mammography films are interpreted using an anatomic approach. A new approach to breast cancer diagnosis utilizes a breast specific gamma camera to measure radiotracer uptake of abnormal tissue in the breast in patients with an abnormal mammogram or palpble mass using technetium sestamibi. Images are taken using the same positioning techniques as mammography for comparison of both types of images. Multi-Institutional trials using a traditional gamma camera demonstrated potential for this approach. However, the inability of traditional gamma camera intrinsic resolution and non-optinuized breast imaging limited scintimammography. Therefore, a breast specific, high resolution gamma camera was developed to overcome these limitations. Results of clinical studies evaluating BSGI are promising and are increasingly being used. Additionally, means for minimally invasive imaging-guided acquisition of tissue are being developed so that biopsies will be avaibles for areas of interest based on radiotracer uptake. Breast specific gamma camera nuclear imaging of the breast is a developing and increasingly utilized approach to improving breast cancer detection and diagnosis.


Developmental trajectories of offending: validation and prediction to young adult alcohol use, drug use, and depressive symptoms.
This longitudinal study extended previous work of Wiesner and Capaldi by examining the validity of differing offending pathways and the prediction from the pathways to substance use and depressive symptoms for 204 young men. Findings from this study indicated good external validity of the offending trajectories. Further, substance use and depressive symptoms in young adulthood (i.e., ages 23-24 through 25-26 years) varied depending on different trajectories of offending from early adolescence to young adulthood (i.e., ages 12-13 through 23-24 years), even after controlling for antisocial propensity, parental criminality, demographic factors, and prior levels of each outcome. Specifically, chronic high-level offenders had higher levels of depressive symptoms and engaged more often in drug use compared with very rare, decreasing low-level, and decreasing high-level offenders. Chronic low-level offenders, in contrast, displayed fewer systematic differences compared with the two decreasing offender groups and the chronic high-level offenders. The findings supported the contention that varying courses of offending may have plausible causal effects on young adult outcomes beyond the effects of an underlying propensity for crime.


Demethylating and anti-hepatocarcinogenic potential of hesperidin, a natural polyphenol of Citrus juices.
Hepatocellular carcinoma (HCC) is a neoplasia representing the fifth most common malignancy worldwide and the third cause of death from cancer. Diets with high content in fruits and vegetables are widely recommended for their health-promoting properties, among them, the protection against diabetes, cancer, and cardiovascular diseases. Hesperidin is the most important phenol in the orange fruit with well-known health benefits. Diet components have been used as possible modulator agents of DNA methylation in cancer cells and epigenetic therapy against their harmful effects could be a potential tool in chemotherapy. The purpose of the present study was to evaluate the methylation patterns induced by hesperidin in HL60 cell line as an in vitro model in order to analyze its chemopreventive effects in epigenetic cancer therapies. A parallel in vivo pilot experience using a rat diethyl nitrosamine hepatocarcinogenesis-induced model was carried out to validate the therapeutic efficacy of this orange flavonol. Results showed that: (i) Hesperidin is cytotoxic in a dose-dependent manner and the IC50 was 12.5 mM; (ii) Hesperidin exerts a significant hypomethylating effect on the LINE-1 sequence (up to 47% hypomethylation at 12.5 mM) and on the ALU-M2 repetitive sequences (up to 32% at 6 mM) in HL60 tumor cells. (iii) Hesperidin does not affect the rat body and liver weight and it is able to reduce the diethyl nitrosamine-induced nodules at 1,000, 500, and 250 ppm. In conclusion, hesperidin could be proposed as a candidate molecule in chemoprevention in epigenetic therapy purposes.


Characterizing degree of lung injury in pediatric acute respiratory distress syndrome.
Although all definitions of acute respiratory distress syndrome use some measure of hypoxemia, neither the Berlin definition nor recently proposed pediatric-specific definitions proposed by the Pediatric Acute Lung Injury Consensus Conference utilizing oxygenation index specify which PaO2/FIO2 or oxygenation index best categorizes lung injury. We aimed to identify variables associated with mortality and ventilator-free days at 28 days in a large cohort of children with acute respiratory distress syndrome. Prospective, observational, single-center study. Tertiary care, university-affiliated PICU. Two-hundred eighty-three invasively ventilated children with the Berlin-defined acute respiratory distress syndrome. None. Between July 1, 2011, and June 30, 2014, 283 children had acute respiratory distress syndrome with 37 deaths (13%) at the Children's Hospital of Philadelphia. Neither initial PaO2/FO2 nor oxygenation index at time of meeting acute respiratory distress syndrome criteria discriminated mortality. However, 24 hours after, both PaO2/FIO2 and oxygenation index discriminated mortality (area under receiver operating characteristic curve, 0.68 [0.59-0.77] and 0.66 [0.57-0.75]; p < 0.001). PaO2/FIO2 at 24 hours categorized severity of lung injury, with increasing mortality rates of 5% (PaO2/FIO2, > 300), 8% (PaO2/FIO2, 201-300), 18% (PaO2/FIO2, 101-200), and 37% (PaO2/FIO2, ≤ 100) across worsening Berlin categories. This trend with 24-hour PaO2/FIO2 was seen for ventilator-free days (22, 19, 14, and 0 ventilator-free days across worsening Berlin categories; p < 0.001) and duration of ventilation in survivors (6, 9, 13, and 24 d across categories; p < 0.001). Similar results were obtained with 24-hour oxygenation index. PaO2/FIO2 and oxygenation index 24 hours after meeting acute respiratory distress syndrome criteria accurately stratified outcomes in children. Initial values were not helpful for prognostication. Definitions of acute respiratory distress syndrome may benefit from addressing timing of oxygenation metrics to stratify disease severity.


Mathematical modeling of the onset of capillary formation initiating angiogenesis.
It is well accepted that neo-vascular formation can be divided into three main stages (which may be overlapping): (1) changes within the existing vessel, (2) formation of a new channel, (3) maturation of the new vessel. In this paper we present a new approach to angiogenesis, based on the theory of reinforced random walks, coupled with a Michaelis-Menten type mechanism which views the endothelial cell receptors as the catalyst for transforming angiogenic factor into proteolytic enzyme in order to model the first stage. In this model, a single layer of endothelial cells is separated by a vascular wall from an extracellular tissue matrix. A coupled system of ordinary and partial differential equations is derived which, in the presence of an angiogenic agent, predicts the aggregation of the endothelial cells and the collapse of the vascular lamina, opening a passage into the extracellular matrix. We refer to this as the onset of vascular sprouting. Some biological evidence for the correctness of our model is indicated by the formation of teats in utero. Further evidence for the correctness of the model is given by its prediction that endothelial cells will line the nascent capillary at the onset of capillary angiogenesis.


Comparison of three tonsillectomy techniques in children.
Tonsillectomy is still one of the most common surgical procedures worldwide performed by otorhinolaryngologists. This single-blind randomized study aimed to compare cold dissection tonsillectomy, coblation tonsillectomy, and harmonic scalpel tonsillectomy in pediatric patients in respect of intraoperative blood loss, operating time, and postoperative pain and bleeding. This single-blind randomized clinical trial evaluated 82 pediatric patients aged 3-16 years (mean age: 7.23 ± 3.26 years) applied with tonsillectomy between April 2017 and March 2020. Harmonic scalpel tonsillectomy was applied to 33 (40.2%) patients, the cold knife technique to 25 (30.5%), and coblation tonsillectomy to 24 (29.3%). There was no statistically significant difference between the three techniques in respect of postoperative pain levels and post-tonsillectomy bleeding rates. The intraoperative bleeding rate and mean operating time were determined to be significantly lower in the harmonic scalpel group (p < 0.05). Harmonic scalpel tonsillectomy is associated with a shorter operating time and lower intraoperative bleeding rates and similar postoperative pain score and postoperative bleeding rates compared with coblation tonsillectomy and cold dissection tonsillectomy. Harmonic scalpel tonsillectomy is a fast, safe, and effective method for tonsillectomy in children.


Isolation, identification and pathogenicity of local entomopathogenic bacteria as biological control agents against the wild cochineal Dactylopius opuntiae (Cockerell) on cactus pear in Morocco.
The Opuntia ficus-indica (L.) cactus, a crucial crop in Morocco, is threatened by the wild cochineal, Dactylopius opuntiae (Cockerell). The aim of this research was to investigate the efficacy of nine bacterial strains against both D. opuntiae nymphs and adults females applied individually or after black soap in the laboratory, greenhouse, and field conditions. Using the partial 16S ribosomal DNA, the bacterial isolates were identified as Pseudomonas koreensis, Pseudomonas sp., Burkholderia sp. and Bacillus sp. Under laboratory conditions, the insecticidal activity of P. koreensis strain 66Ms.04 showed the level mortality (88%) of adult females' at 108 CFU/mL, 7 days after application. At a concentration of 108 CFU/mL, P. koreensis strain 66Ms.04 and Pseudomonas sp. (strains 37 and 5) caused 100% nymphs mortality rate three days after application. Under greenhouse conditions, the use of P. koreensis strain 66Ms.04 at 108 CFU/mL following the application of black soap (60 g/L) demonstrated the maximum levels of females and nymphs' mortalities with 80 and 91.25%, respectively, after 8 days of treatment. In field conditions, the combined application of the P. koreensis strain 66Ms.04 at 108 CFU/mL with black soap at 60 g/L, for an interval of 7 days, significantly increased the mortality of adult females to 93.33% at 7 days after the second application. These findings showed that the combined treatment of P. koreensis strain 66Ms.04 with black soap can be a potent and eco-friendly pesticide against D. opuntiae.


Chronic hepatitis B: Advances in treatment.
Treatment of chronic hepatitis B (CHB) has markedly improved in the last 15 years due to the availability of direct antivirals which greatly increase therapeutic options. Currently, there are two classes of agents licensed for CHB treatment: standard or pegylated interferon alpha (IFN or Peg-IFN) and five nucleoside/nucleotide analogues (NAs). Long-term treatment with NAs is the treatment option most often used in the majority of CHB patients. Entecavir and tenofovir, the most potent NAs with high barrier to resistance, are recommended as first-line monotherapy by all major treatment guidelines and can lead to long-lasting virological suppression, resulting in histological improvement or reversal of advanced fibrosis and reduction in disease progression and liver-related complications. In this review, we focus on current treatment strategies of chronic hepatitis B and discuss the most recent efficacy and safety data from clinical trials and real life clinical practice. Recent findings of response-guided approaches are also discussed.