Food consumption patterns in preschool children.
Healthy eating during early childhood is important for growth and development. Eating Well with Canada's Food Guide (CFG) provides dietary recommendations. We investigated patterns of food consumption among preschool children and attempted to determine whether these children's intakes met nutrition recommendations. Between 2005 and 2007, four- and five-year-old children (n=2015) attending 12 Edmonton-region public health units for immunization were recruited for a longitudinal study on determinants of childhood obesity. The children's dietary intake at baseline was assessed using parental reports. Overall, 29.6%, 23.5%, 90.9%, and 94.2% of the children met recommendations for vegetables and fruit, grain products, milk and alternatives, and meat and alternatives, respectively. In addition, 79.5% consumed at least one weekly serving of foods in the "choose least often" group. Significant differences existed in consumption of food groups across socioeconomic and demographic groups. For example, 82.9%, 84.7%, and 75.9% of preschool children from neighbourhoods of low, medium, and high socioeconomic status, respectively, consumed at least one food in the "choose least often" group (χ² =16.2, p<0.001). Consumption of vegetables and fruit and grain products was low among participants, and intake of "choose least often" foods was high. Consumption of foods also differed among socioeconomic and demographic groups. To encourage healthy eating among children, public health professionals should target groups who do not meet the CFG recommendations.


Diagnosis of aggressive periodontitis: A dilemma?
Periodontitis is one of the leading causes of tooth loss in the adult population. This disease can be classified into various categories, and one of the most destructive amongst them is aggressive periodontitis (AgP). The incidence of AgP is lower than other types of periodontitis. However, it affects young individuals and can cause severe destruction of tooth-supporting structures including tooth loss if left untreated. The current classification for diagnosing periodontal disease was established by the American Academy of Periodontology (AAP) in 1999. This classification provided strict guidelines to aid in AgP diagnosis. These include three main factors: systemically healthy individual, rapid loss of clinical attachment, and familial aggregation. In spite of these specific guidelines, AgP diagnosis is often missed clinically due to various reasons. There is still a vast variation in the diagnostic criteria for identifying AgP and not all practitioners utilize the AAP guidelines for their diagnosis. Furthermore, the definition of the disease might be changing in the future to better represent the current understanding of the disease. Since early diagnosis and prompt treatment is key in treating these patients, it is important to have calibration in the diagnosis process. This review aims to identify sources of variation and ambiguity in diagnosing AgP among dental practitioners. For this purpose, we have conducted an extensive literature search and outlined the various diagnostic aids for AgP patients reported in the literature. Understanding and correcting these variations can simplify the diagnostic process leading to faster treatment of patients affected with AgP. This review also emphasizes the importance of minimizing the bias in identifying patients with AgP and highlights the best tools for this purpose.


Primary left atrial leiomyosarcoma: literature review and lessons of a case.
Primary cardiac sarcoma is an uncommon disease. In particular, leiomyosarcoma of cardiovascular origin is extremely rare. Half of all cardiac leiomyosarcomas are located in the left atrium. Due to the extreme rarity of left atrial leiomyosarcoma, there is no great experience in its management. This review includes a report of a case of left atrial leiomyosarcoma followed up over 45 months. The literature review examines the distribution of left atrial leiomyosarcoma, the physiological reasons for the tendency of cardiac leiomyosarcoma to be localized to the left atrial cavity, the clinical and physical appearance of this disease, and the key differences between left atrial leiomyosarcoma and the most common left atrial tumor, myxoma. The morphological features, using light and electron microscopy and immunochemical staining, are discussed. Treatment modalities including adjuvant therapy and surgical resection are examined and their effectiveness compared. Opinions regarding the results and optimal treatment of leiomyosarcoma are not always in agreement. This highlights the need for inter-hospital comparison to determine the optimal treatment regimen.


Lytic infection of Epstein-Barr virus (EBV) in hemophagocytic syndrome associated with EBV-induced lymphoproliferative disorder.
This report describes lytic infection with Epstein-Barr virus (EBV) in three cases of hemophagocytic syndrome (HPS) presumably associated with EBV-induced lymphoproliferative disorder. All cases were previously healthy females with ages ranging from 52 to 87 years who showed a fulminant clinical course with accompanying fever, liver dysfunction, and disseminated intravascular coagulation. Cases 1 and 2 showed proliferation of atypical T lymphocytes, and case 3 showed proliferation of atypical B lymphocytes. Hemophagocytic histiocytes were observed in the bone marrow, lymph nodes, spleen, and liver. Atypical lymphocytes in all cases showed a positive reaction for both EBV-encoded small RNA (EBER) indicating latent infection with EBV and immediate early mRNAs of the Bam HI fragment of lower stranded frame (BHLF), indicating lytic infection by in situ hybridization. Interestingly, BHLF-positive cells were predominant in all cases. It is possible that reactivation of EBV infection may be involved in triggering the induction of cytokines and abnormal activation of histiocytes.


The application of cost effectiveness analysis to derive a formulary for urinary tract infections.
According to economic principles an inappropriate prescription is the choice of an antimicrobial with higher/equivalent cost and lower effectiveness (or higher cost and equivalent/lower efficacy) than an alternative (in this case, the former is specified as a "dominated" drug). To identify cost-effective antibiotics we applied the principles of incremental cost-effectiveness analysis (ICEA) to microbiological data of San Bortolo Hospital. Its 27 wards were grouped in 9 functional areas. The resistance patterns of 8 urinary pathogens in the 1997 microbiology data base were assessed. The measure of antibiotic effectiveness was expressed as the percentage of isolates susceptible to each antibiotic tested. The difference in cost (i.e. the incremental change) between each antibiotic and the next more expensive alternative was calculated, and compared with the incremental change in effectiveness. Calculations were made for each pathogen. The antibiotics remaining after exclusion of all "dominated" antibiotics were pooled on a list defined as "Specific Area Formulary". The implications of the use of economic principles within a general antimicrobial policy are discussed.


Intratumoral delivery of RIG-I agonist SLR14 induces robust antitumor responses.
Cytosolic nucleic acid-sensing pathways can be triggered to enhance immune response to cancer. In this study, we tested the antitumor activity of a unique RIG-I agonist, stem loop RNA (SLR) 14. In the immunogenic tumor models, we observed significant tumor growth delay and an extended survival in SLR14-treated mice. SLR14 also greatly improved antitumor efficacy of anti-PD1 antibody over single-agent treatment. SLR14 was mainly taken up by CD11b+ myeloid cells in the tumor microenvironment, and many genes associated with immune defense were significantly up-regulated after treatment, accompanied by increase in the number of CD8+ T lymphocytes, NK cells, and CD11b+ cells in SLR14-treated tumors. Strikingly, SLR14 dramatically inhibited nonimmunogenic B16 tumor growth, and the cured mice developed an immune memory. Furthermore, a systemic antitumor response was observed in both bilateral and tumor metastasis models. Collectively, our results demonstrate that SLR14 is a promising therapeutic RIG-I agonist for cancer treatment, either alone or in combination with existing immunotherapies.


Molecular mechanisms of bacterial pathogenicity.
Cautious optimism has arisen over recent decades with respect to the long struggle against bacteria, viruses, and parasites. This has been offset, however, by a fatal complacency stemming from previous successes such as the development of antimicrobial drugs, the eradication of smallpox, and global immunization programs. Infectious diseases nevertheless remain the world's leading cause of death, killing at least 17 million persons annually [61]. Diarrheal diseases caused by Vibrio cholerae or Shigella dysenteriae kill about 3 million persons every year, most of them young children: Another 4 million die of tuberculosis or tetanus. Outbreaks of diphtheria in Eastern Europe threatens the population with a disease that had previously seemed to be overcome. Efforts to control infectious diseases more comprehensively are undermined not only by socioeconomic conditions but also by the nature of the pathogenic organisms itself; some isolates of Staphylococcus aureus and Enterobacter have become so resistant to drugs by horizontal gene transfer that they are almost untreatable. In addition, the mechanism of genetic variability helps pathogens to evade the human immune system, thus compromising the development of powerful vaccines. Therefore detailed knowledge of the molecular mechanisms of microbial pathogenicity is absolutely necessary to develop new strategies against infectious diseases and thus to lower their impact on human health and social development.


Radiofrequency Ablation for Stage I Non-Small Cell Lung Cancer: An Updated Review of Literature from the Last Decade.
This review summarizes the current findings on radiofrequency ablation (RFA) for stage I non-small cell lung cancer (NSCLC) from relevant literature published in the last decade. While most earlier studies included small populations and had short follow-up periods, more robust data have become available owing to prospective or large cohort studies. The reported overall survival rates after RFA for stage I NSCLC were 83-96%, 40-74%, and 23-61% at 1, 3, and 5 years, respectively, in recent studies. Furthermore, many comparative studies on the outcomes of RFA and stereotactic body radiotherapy have been performed. Most of these studies report no significant difference in survival outcomes between the therapies. Currently, major guidelines define RFA as a reasonable alternative treatment for stage I NSCLC in non-surgical candidates.


The prevalence of interdental gingival inflammation: a report from the 1986 ADA annual health screening.
This study screened 1986 ADA annual session participants for interdental gingival inflammation by using the Eastman Interdental Bleeding Index (EIBI). The EIBI provides a sensitive and objective measure of interdental gingival inflammation, and has been shown to be reliable. At least one bleeding site was found in 81% of the 435 dentists examined. This study showed that in a dental population, the prevalence of gingival inflammation was high when the health of interdental tissues was assessed.


Allergenic properties of roasted peanut allergens may be reduced by peroxidase.
Peanut allergy is a public health issue. The culprits are the peanut allergens. Reducing the allergenic properties of these allergens or proteins will be beneficial to allergic individuals. In this study, the objective was to determine if peroxidase (POD), which catalyzes protein cross-linking, reduces the allergenic properties of peanut allergens. In the experiments, protein extracts from raw and roasted defatted peanut meals at pH 8 were incubated with and without POD in the presence of hydrogen peroxide at 37 degrees C for 60 min. The POD-treated and untreated samples were then analyzed by SDS-PAGE, western blots, and competitive inhibition ELISA. IgE binding or allergenicity was determined in blots and ELISA. Results showed that POD treatment had no effect on raw peanuts with respect to protein cross-linking. However, a significant decrease was seen in the levels of the major allergens, Ara h 1 and Ara h 2, in roasted peanuts after POD treatment. Also, polymers were formed. Despite this, a reduction in IgE binding was observed. It was concluded that POD induced the cross-linking of mainly Ara h 1 and Ara h 2 from roasted peanuts and that, due to POD treatment, IgE binding was reduced. The finding indicates that POD can help reduce the allergenic properties of roasted peanut allergens.


Stimulation of mouse lymphocytes by a mitogen derived from Mycoplasma arthritidis. VII. Responsiveness is associated with expression of a product(s) of the V beta 8 gene family present on the T cell receptor alpha/beta for antigen.
Mycoplasma arthritidis produces a potent soluble T lymphocyte mitogen (MAM) which is dependent upon accessory cells bearing the alpha-chain of the I-E molecule (E alpha). Lymphocytes from the RIIIS mouse strain which possess E alpha yet whose T cells fail to recognize the MAM-E alpha complex were shown not to express V beta 8.1, 8.2 or 8.3 gene products present on the TCR-alpha/beta by virtue of their lack of reactivity with the F23.1 mAb. Because lymphocytes from the congenic B10.RIII mouse strain reacted positively with F23.1, we examined the progeny from (RIIIS x B10.RIII)F1 x RIIIS backcross mice for cosegregation of lymphocytes expressing F23.1-reactive sites and ability to proliferate in response to MAM. Whereas lymphocytes of all mice responded to Con A, only lymphocytes from progeny expressing F23.1-reactive cells responded to MAM. Similar studies were conducted on progeny from the F23.1- SWR mouse which was backcrossed to (SWR x B10.RIII)F1 mice. Of the E alpha-bearing progeny, there was a direct correlation between lymphocyte expression of F23.1 determinants and ability to respond to MAM. These results established that MAM reactivity was dependent upon a product(s) of the V beta locus of the TCR-alpha/beta. Nodal and thymic lymphocytes cultured for 3 days in vitro with MAM exhibited clonal expansion of F23.1 and F23.2-reactive cells as compared with cultures treated with Con A. We also demonstrated that F23.1 and F23.2 mAb inhibited the ability of lymphocytes to proliferate in response to MAM but had little effect on responses to Con A. The combined data suggest that the MAM-E alpha complex can utilize a V beta 8 gene product(s) on the TCR-alpha/beta.


Stereotactic CT scanning for the biopsy of intracranial lesions and functional neurosurgery.
This report describes a system for incorporation of stereotactic CT scanning data, stereotactic arteriographic data and a computer-generated stereotactic atlas into a three-dimensional matrix utilizing an operating room computer. 86 patients have undergone computer-assisted stereotactic biopsies of intracranial lesions without mortality or neurologic morbidity. Neuroablative and neuroaugmentative procedures have been performed on 5 patients using the CT stereotactic atlas with good correlation with target points determined by ventriculography and microelectrode recording.


[Attitudes of medical students towards psychotherapeutic treatment and training].
Against the background of insufficient medical care for patients with psychiatric and psychosomatic disorders it is important to ascertain the interests and attitudes of medical students towards mental disorders, psychotherapeutic treatment and training. Medical students (N = 366) in their clinical training were given half-standardized questionnaires about their attitudes towards psychotherapeutic treatments, their career plans, psychosocial distress and sociodemographic characteristics. There was an overall high interest in psychological distress of patients and in psychotherapeutic treatment methods. Contrary to these findings, however, only few medical students were willing to specialize in psychosomatic medicine, psychiatry or psychotherapy. Psychosomatic and psychotherapeutic career choices were more frequent in women as well as in students who had experienced psychological distress themselves, and who had undergone psychotherapy, and who were less oriented toward economic success and careers. Medical students regarded psychological characteristics as generally interesting and important for medical care, but rarely considered specializing in these medical fields after graduation. Potential remedies (e.g., improved teaching of psychosocial competences and psychotherapeutic knowledge, career counselling) are discussed.


To what extent is telehealth reported to be incorporated into undergraduate and postgraduate allied health curricula: A scoping review.
Telehealth has become a necessity within the medical and allied health professions since the COVID-19 Pandemic generated a rapid uptake worldwide. It is now evident that this health delivery format will remain in use well into the future. However, health education training, most particularly allied health, has been slow to 'catch up' and adapt curriculum to ensure graduates are equipped with the knowledge and skills to implement telehealth in the workplace. The aim of this study was to gain a comprehensive understanding of current telehealth curricula in undergraduate and postgraduate allied health education training programs, with a focus on the aims, objectives, content, format, delivery, timeline and assessments. A systematic search of Medline, Embase, PsychINFO, Scopus, ERIC and relevant grey literature was conducted. Students studying allied health degrees through formal education at either postgraduate or undergraduate level were included, while nursing, dentistry and medical students were excluded. The data from the included studies was extracted and tabulated by country, participants, program and content. Of the 4484 studies screened, eleven met the eligibility criteria. All studies were published after 2012, highlighting the recency of research in this area. The studies were conducted in four countries (Australia, United Sates of America, United Kingdom, Norway) and participants were from various allied health professions. Of the included studies, four related to undergraduate programs, four to postgraduate programs and for the remaining three, this was not specified. Curricula were delivered through a combination of online and face-to-face delivery, with assessment tasks, where reported, comprising mainly multiple-choice and written tests. Published reporting of telehealth curricula within allied health programs is limited. Even the minority of programs that do include a telehealth component lack a systematic approach. This indicates that further primary research would be beneficial in this area.


Inhibition of PI3K/AKt/mTOR signaling pathway protects against d-galactosamine/lipopolysaccharide-induced acute liver failure by chaperone-mediated autophagy in rats.
This study aims to investigate the effects of PI3K/AKt/mTOR signaling pathway on the proliferation and apoptosis in acute liver failure (ALF) by chaperone mediated autophagy (CMA). The hepatocytes extracted from both normal rats and rats with ALF were assigned to control, acute injury, P13K agonist, and P13K inhibitor groups. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used as part of this investigation to detect the expression of PI3K/AKt/mTOR signaling pathway related-proteins (PI3K, AKt, mTOR), apoptosis related-proteins (Fas, Bax, Bcl-2), chaperone-mediated autophagy (CMA) marker proteins (LAMP-2A, HSC 70), p-PI3K, p-AKt, p-4E-BPI, and p-S6K. An MTT assay was used for analysis of cell proliferation after transfection. Flow cytometry is performed to detect the cell apoptosis. In comparison to the normal group, the model group showed enhanced positive rate of PI3K, AKt, mTOR, increased expression levels of PI3K, AKt, mTOR, Fas, Bax, p-PI3K, p-AKt, p-4E-BPI and p-S6K, reduced expression levels of Bcl-2, LAMP-2A and HSC 70. The results in vitro experiment: compared with the acute injury group, the PI3K agonist group showed elevated expression levels of PI3K, AKt, mTOR, Fas, Bax, p-PI3K, p-AKt, p-4E-BPI and p-S6K, decreased expression levels of Bcl-2, LAMP-2A and HSC 70, inhibited cell proliferation, more arrested cells in G1 stage, and promoted cell apoptosis. Opposing this, the P13K inhibitor group exhibited an opposite trend. In conclusion, inhibition of the PI3K/AKt/mTOR signaling pathway plays a protective role in ALF by promoting CMA expression, which could arrest cell proliferation and promote cell apoptosis.


Hyperexcitability of the CA1 hippocampal region during epileptogenesis.
Temporal Lobe Epilepsy (TLE) is often preceded by a latent (seizure-free) period during which complex network reorganizations occur. In experimental epilepsy, network hyperexcitability is already present during the latent period, suggesting a modification of information processing. The purpose of this study was to assess the input/output relationship in the hippocampal CA1 region during epileptogenesis. Field recordings in strata pyramidale and radiatum were used to measure the output of CA1 pyramidal cells as a function of the synaptic inputs they receive following the stimulation of Shaffer collaterals in slices obtained from sham and pilocarpine-treated animals during the latent and chronic periods. We show that there is a transient increase of the input and output field responses during the latent period as compared to sham and epileptic animals. The coupling between excitatory inputs and cell firing was also increased during the latent period. This increase persisted in epileptic animals, although to a lesser extent. We also confirm that paired-pulse facilitation occurs before the chronic phase. The present data further support the view that hyperexcitability is present at an early stage of epileptogenesis. Network output is more facilitated during the latent than during the chronic period. Hyperexcitability may participate to epileptogenesis, but it is not sufficient in itself to produce seizures.


Role of nitric oxide in benzodiazepines-induced antinociception in mice.
The influence of nitric oxide (NO) on antinociceptive activity of diazepam (DZ), chlordiazepoxide (CDP) and clonazepam (CZ) was examined using the writhing test in mice. The effect of DZ was also studied in mice using hot plate and tail flick tests. DZ (1.25, 2.5 and 5 mg/kg), CDP (1.25, 2.5, 5, 10 and 20 mg/kg) and CZ (0.075, 0.3125, 0.625, 1.25 and 2.5 mg/kg) produced significant, dose-dependent (DZ, CDP) antinociception in mice. The benzodiazepines (BZs)-induced antinociception was antagonized by flumazenil (5 mg/kg) and was not changed by naloxone (2.5, 5 and 10 mg/kg), except that of CZ, which was reversed by 5 mg/kg of naloxone. NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) as well as 7-nitroindazole (7-NI) intensified antinociceptive activity of BZs. The antinociceptive effect resulting from co-administration of L-NAME with CZ and 7-NI with CDP was reversed by L-arginine. Methylene blue (MB) increased, whereas L-arginine (but not D-arginine) decreased antinociceptive effects of the studied BZs. These results suggest that the NO-cGMP pathway is involved in the mechanism of BZs-induced antinociception in the writhing test in mice.


Genetic counselling: its need in psychiatry and the directions it gives for future research.
The methods for investigating the extent to which genetic factors can influence vulnerability to psychiatric illness are, in increasing order of precision: family, twin, and adoption studies. The evidence from these studies is in support of a gene-environment interaction for schizophrenia and the affective disorders. While the family study method cannot supply precise etiological data, the empirically derived information can be used by the genetic counsellor to provide empirical risk estimates to the counsellee. The psychiatrist, geneticist, and social worker make an appropriate team for reliable genetic counselling. The clinician must determine the precise psychiatric diagnoses in family members which the geneticist may use to estimate risk. The social worker can follow-up the counselling session or sessions to assess the counsellees' understanding of what has been told to them. It is stressed that while genetic counselling should be available, clinical judgement should be exercised to ensure its appropriate use.


Detection and Identification of Six Monilinia spp. Causing Brown Rot Using TaqMan Real-Time PCR from Pure Cultures and Infected Apple Fruit.
Brown rot is a severe disease affecting stone and pome fruit. This disease was recently confirmed to be caused by the following six closely related species: Monilinia fructicola, M. laxa, M. fructigena, Monilia polystroma, M. mumecola, and M. yunnanensis. Because of differences in geographic distributions, some of these species are important quarantine pathogens in certain countries. In this study, we developed TaqMan real-time polymerase chain reaction (PCR) assays to detect and identify the six species. Primer pairs and probes were designed for Monilinia fructicola, M. fructigena, M. laxa, and Monilia polystroma based on sequence differences in the laccase-2 genes. Additionally, based on sequence differences in the elongation factor genes, primer pairs and probes were designed for Monilia mumecola and M. yunnanensis. The real-time PCR assays were able to specifically identify the target pathogens, with detection limits of 10 to 100 fg of DNA, which is equivalent to one to seven conidia. The assays were also able to detect the target pathogens in a mixed DNA sample comprising all six Monilinia spp. and related species. The real-time PCR assays accurately detected target fungi from infected apple fruit. Furthermore, the identification results were consistent with those of traditional morphological methods.


Cooperative interactions in hybrids of aspartate transcarbamylase containing succinylated regulatory polypeptide chains.
Succinylated derivatives of the regulatory subunit of aspartate transcarbamylase of Escherichia coli were prepared by treating the intact enzyme with succinic anhydride followed by dissociation of the modified protein into catalytic and regulatory subunits which were separated by ion exchange chromatography. The succinylated regulatory subunits were used in hybridization experiments with native subunits to study the organization of the six regulatory polypeptide chains in the intact enzyme. Rapid mixing of succinylated and native regulatory subunits with native catalytic subunits yielded a four-membered hybrid set of reconstituted enzyme-like molecules; hence, the assembly process involves three regulatory combining units and the six regulatory polypeptide chains in the intact enzyme must be arranged as three dimeric subunits. When the modified and native regulatory subunits were incubated together for only brief periods (less than 1 min) followed by the addition of catalytic subunits, the resulting hybrid set was complex with no resolution of discrete species. Apparently the isolated regulatory dimers dissociate readily and reversibly into single polypeptide chains due to relatively weak intra-subunit bonding domains. In contrast, after reconstitution of enzyme-like molecules, the incorporated succinylated regulatory subunits did not exchange with free subunits. Enzyme-like molecules containing three extensively succinylated regulatory subunits show reduced binding of the inhibitor, CTP, and lack both the homotropic and heterotropic effects characteristic of native aspartate transcarbamylase. Preparations containing only slightly succinylated regulatory subunits showed only little inhibition by CTP and considerable cooperativity. The decrease in homotropic effects in these reconstituted molecules correlated with the reduction in the succinate-promoted change in the sedimentation coefficient. Reconstituted enzyme-like molecules containing regulatory subunits which had been extensively succinylated in the presence of CTP retained their binding capacity even though they were only slightly inhibited by CTP and exhibited reduced cooperativity. Hybrid molecules containing both native and succinylated regulatory subunits also possessed reduced allosteric behavior.


Mediating positive moods: the impact of experiencing compassion at work.
The aim of this study was to examine the mediating effects of positive moods underlying the relationship between experiencing compassion at work and both job performance and organisational citizenship behaviour (OCB). Employees who have experienced compassion might have more positive moods while at work; such moods have been connected to a host of important organisational outcomes such as better job performance and OCB. This study adopted a two-stage survey whose participants consisted of 269 registered nurses employed by hospitals in Taiwan. All hypotheses were tested using hierarchical regression analyses. The results showed that positive moods mediate completely the relationship between experiencing compassion at work and both job performance and OCB. When nurses frequently feel that their hospital, supervisors or co-workers are willing to listen to them, the strength that arises from this compassion can effectively increase job performance and OCB though the positive moods process. It is importance to cultivate a compassionate and caring culture throughout hospitals' hierarchies, especially from the top down. Nurses will have more opportunities to experience it, witness it, or participate in it when they work within a compassionate and caring organisational culture.


Autophagic cell death: Loch Ness monster or endangered species?
The concept of autophagic cell death was first established based on observations of increased autophagic markers in dying cells. The major limitation of such a morphology-based definition of autophagic cell death is that it fails to establish the functional role of autophagy in the cell death process, and thus contributes to the confusion in the literature regarding the role of autophagy in cell death and cell survival. Here we propose to define autophagic cell death as a modality of non-apoptotic or necrotic programmed cell death in which autophagy serves as a cell death mechanism, upon meeting the following set of criteria: (i) cell death occurs without the involvement of apoptosis; (ii) there is an increase of autophagic flux, and not just an increase of the autophagic markers, in the dying cells; and (iii) suppression of autophagy via both pharmacological inhibitors and genetic approaches is able to rescue or prevent cell death. In light of this new definition, we will discuss some of the common problems and difficulties in the study of autophagic cell death and also revisit some well-reported cases of autophagic cell death, aiming to achieve a better understanding of whether autophagy is a real killer, an accomplice or just an innocent bystander in the course of cell death. At present, the physiological relevance of autophagic cell death is mainly observed in lower eukaryotes and invertebrates such as Dictyostelium discoideum and Drosophila melanogaster. We believe that such a clear definition of autophagic cell death will help us study and understand the physiological or pathological relevance of autophagic cell death in mammals.


The evolution of genetic-based risk scores for lipids and cardiovascular disease.
With improved next-generation sequencing technology, open-access genetic databases and increased awareness of complex trait genetics, we are entering a new era of risk assessment in which genetic-based risk scores (GRSs) will play a clinical role. We review the concepts underlying polygenic models of disease susceptibility and challenges in clinical implementation. Polygenic risk scores are currently used in genetic research on dyslipidemias and cardiovascular disease (CVD). Although the underlying principles for constructing polygenic scores for lipids are established, the lack of consensus on which score to use is indicated by the large number - about 50 - that have been published. Recently, large-scale polygenic scores for CVD appear to afford superior risk prediction compared to small-scale scores. Despite the potential benefits of GRSs, certain biases towards ethnicity and sex need to be worked through. We are on the verge of clinical application of GRSs to provide incremental information on dyslipidemia and CVD risk above and beyond traditional clinical variables. Additional work is required to develop a consensus of how such scores will be constructed and measured in a validated manner, as well as clinical indications for their use.


Aging and platelet beta-adrenoceptor function.
Ten young volunteers (mean age 20 years) and ten elderly volunteers (mean age 89 years), including equal numbers of healthy men and women, were tested regarding adrenaline-induced platelet aggregation in vitro, platelet cyclic AMP content before and after beta-adrenoceptor stimulation with isoprenaline, as well as binding by platelet membranes of 125I-hydroxybenzylpindolol. In the aged patients there was a highly significant decrease in the concentration of adrenaline needed to produce irreversible platelet aggregation, as compared with the young. Platelet basal cyclic AMP content did not differ but the response to isoprenaline stimulation, expressed as the percentage rise of cyclic AMP content above the unstimulated level, was significantly decreased in the old subjects. The beta-adrenoceptor number was unchanged but the affinity decreased significantly in the old group. The data suggest that the increased aggregation response to adrenaline in old people could be due in part to a diminished functional capacity of the platelet beta-adrenoceptor.


Disseminated cerebral hydatid disease (multiple intracranial echinococcosis).
Intracranial echinococcosis is relatively uncommon and usually occurs in the context of echinococcal lesions elsewhere in the body, mostly liver and lung. Multiple intracranial lesions can result from rupture of an initial single intracranial cyst (in cystic echinococcosis) or from dissemination of systemic disease of the lung, liver or heart (cystic and alveolar echinococcosis). The two main subtypes, cystic and alveolar echinococcosis, present differently and have distinct imaging features in the brain. We discuss the presentation, imaging findings and clinical course of three cases (two cystic and one alveolar) of intracranial echinococcal disease in adults.


Postmortem radiography after unexpected death in neonates, infants, and children: should imaging be routine?
The purpose of this study was to determine whether postmortem radiography of neonates, infants, and children provides additional information that is not detected at autopsy in cases of unexpected death. Inclusion criteria for 106 consecutive postmortem skeletal surveys (1998-2000) were neonates, infants, and children 2 years old or younger with no preexisting medical condition to account for mortality. Pediatric radiologists interpreted all the radiographic examinations, which consisted of high-detail, collimated anteroposterior radiographs of the appendicular and axial skeleton, lateral radiographs of the axial skeleton, and oblique radiographs of the ribs. Imaging results were compared with those obtained from standard protocol autopsies on all children. Four categories of death were designated: homicide (i.e., abuse, n = 14), accidental (e.g., drowning, n = 28), natural (e.g., acute illness, n = 43), and undetermined (n = 21). The causes of death in the 14 child abuse victims were blunt force injuries to the intracranial (n = 11) and chest and abdominal (n = 1) areas; asphyxia (n = 1); and shaking injury (n = 1). In six (43%) of these 14 patients, radiography detected 26 extremity fractures that had not been detected at autopsy; four (67%) of these six patients had fractures of different ages that involved more than one extremity. All fractures carried a high index of suspicion of abuse. No skeletal injuries were found in cases of accidental, undetermined, and natural deaths. Postmortem radiography provides important additional information regarding the extent and chronicity of extremity trauma that may not be documented at autopsy. This finding supports the routine use of radiography in cases of suspected child abuse. Normal findings on postmortem skeletal radiography may help to distinguish cases of natural, accidental, and undetermined causes of death from those of abuse, aiding in the proper handling of these cases by medical and law enforcement personnel.


[Investigation of trend in patients with pollinosis using web site: Comparison of these three years].
Recently, the number of patient with Japanese cedar pollinosis (JCP) has been increasing. In the last 3 years, the amount of pollen has been very volatile. In this study, we examined that whether the amount of pollen influenced symptoms and selfcare using web site. For this examination, the questionnaire on web site was used from March 15 to April 30, 2003, 2004, and 2005 respectively. The rate of early treatments was high in the years when the amount of pollen was large, while the rate was low in the years when the amount of pollen was little. In the comparison of symptoms, the severity of the symptoms, in particular, nasal congestion, itchy eyes, watering eyes, and disturbance of sleep, was highly related to the amount of pollen. In addition, these symptoms were the severest in 2005. And the selfcare was also changing according to the amount of pollen. We concluded the greater the amount of pollen was, the severer the nasal congestion, eye symptoms, and disturbance of sleep were. The questionnaire survey that uses web site is a limited investigation of patient who has the access to the Internet. However, in the trend survey of JCP patients, it seemed that was extremely meaningful.


[Relaxing effects of local heat-air exposure under stresses in students].
The relaxing effects of local heat-air exposures (TAE) on man (60 male students aged 19-21 years) were studied using a specially designed device which may control temperatures and time on an individual basis depending on the examinees' sensation of a positive emotion. On study days and under examination stress. TAE was shown to reduce situation-induced anxiety, to increase the duration and variability of R-R intervals on ECG, to make EEG right-frontal delta- and theta-indices lower and EEG right occipital and frontal alpha-index higher. The range of visual accommodation increased. The findings suggest that the proposed graded local heat-air exposure procedure is an effective tool of relaxation and prevention of negative effects of emotional stress.


Total deletion of in vivo telomere elongation capacity: an ambitious but possibly ultimate cure for all age-related human cancers.
Despite enormous effort, progress in reducing mortality from cancer remains modest. Can a true cancer "cure" ever be developed, given the vast versatility that tumors derive from their genomic instability? Here we consider the efficacy, feasibility, and safety of a therapy that, unlike any available or in development, could never be escaped by spontaneous changes of gene expression: the total elimination from the body of all genetic potential for telomere elongation, combined with stem cell therapies administered about once a decade to maintain proliferative tissues despite this handicap. We term this therapy WILT, for whole-body interdiction of lengthening of telomeres. We first argue that a whole-body gene-deletion approach, however bizarre it initially seems, is truly the only way to overcome the hypermutation that makes tumors so insidious. We then identify the key obstacles to developing such a therapy and conclude that, while some will probably be insurmountable for at least a decade, none is a clear-cut showstopper. Hence, given the absence of alternatives with comparable anticancer promise, we advocate working toward such a therapy.


Immunological repertoire linked to PSTPIP1-associated myeloid-related inflammatory (PAMI) syndrome.
Mutations along PSTPIP1 gene are associated to two specific conditions, PAPA syndrome and PAMI syndrome, both autoinflammatory disorders associated to disturbances in cytoskeleton formation. Immunological aspects of PAMI syndrome has not yet been reported neither the clinical impact on therapeutical decisions. Clinical data of patients records were retrospectively accessed. Genomic DNA were extracted and sequenced following standard procedures. Peripheral lymphocytes were quantified in T, B e FOXP3 phenotypes. We describe two related patients with PAMI syndrome harboring the usual E250K mutation. Anti-IL1 therapy could partially control the disease in the index patient. A broad spectrum of immunological effects as well as an aberrant expression of FOXP3 could be observed. Here we report two related brazilian patients with PAMI syndromes harboring the E250K mutation in PSTPIP1, their immunological aspects and the therapeutical response to canakinumab.


Long-term efficacy and safety of combined prolonged-release oxycodone and naloxone in the management of non-cancer chronic pain.
The aim of this study was to assess safety and efficacy of fixed combination oxycodone prolonged release (PR)/naloxone PR in terms of both analgesia and improving opioid-induced bowel dysfunction (OIBD) and associated symptoms, such as opioid-induced constipation (OIC), in adults with chronic non-cancer pain. These were open-label extension studies in which patients who had previously completed a 12-week, double-blind study received oxycodone PR/naloxone PR for up to 52 weeks. The analgesia study assessed pain using the modified Brief Pain Inventory-Short Form (BPI-SF). The bowel function study assessed improvements in constipation using the Bowel Function Index (BFI). At open-label baseline in the analgesia study (n = 379), mean score [+/- standard deviation (SD)] for the BPI-SF item 'average pain over the last 24 h' was 3.9 +/- 1.52, and this remained low at 6 months (3.7 +/- 1.59) and 12 months (3.8 +/- 1.72). Mean scores for BPI-SF item 'sleep interference', and the BPI-SF 'pain' and 'interference with activities' subscales also remained low throughout the 52-week study. In the bowel function study (n = 258), mean BFI score (+/- SD) decreased from 35.6 +/- 27.74 at the start of the extension study to 20.6 +/- 24.01 after 12 months of treatment with oxycodone PR/naloxone PR. Pain scores also remained low and stable during this study. Adverse events in both extension phases were consistent with those associated with opioid therapy; no additional safety concerns were observed. Results from these two open-label extension studies demonstrate the long-term efficacy and tolerability of fixed combination oxycodone PR/naloxone PR in the treatment of chronic pain. Patients experienced clinically relevant improvements in OIBD while receiving effective analgesic therapy.


Results of a randomized, double-blind, vehicle-controlled efficacy trial of pimecrolimus cream 1% for the treatment of moderate to severe facial seborrheic dermatitis.
Seborrheic dermatitis is commonly treated with anti-inflammatory products, including topical corticosteroids. Pimecrolimus cream 1% also exerts anti-inflammatory activity by inhibiting T-cell cytokine production. We sought to compare the efficacy and safety of twice-daily pimecrolimus for treatment of moderate to severe facial seborrheic dermatitis. This double-blind, vehicle-controlled, 4-week trial randomized patients with seborrheic dermatitis to pimecrolimus or vehicle (1:1). Clinical assessments (erythema [0-3] and scaling [0-3] combined for a total area score [0-6]) were performed at weeks 0, 2, and 4. Inclusion criteria included total area score 4 or greater and erythema 2 or greater. The prespecified primary variable, change from baseline in total area score at week 4, was analyzed using a two-sample t test for intent-to-treat and per protocol populations. In all, 96 adults of mean age 59.6 years, 88.5% male, were randomized (n = 47 pimecrolimus; 49 vehicle). At week 4, the mean change from baseline in total area score was 3.7 versus 3.3 for pimecrolimus and vehicle groups, respectively (intent-to-treat: P = .1913; 95% confidence interval (CI) for difference [-0.195, 0.961]). Per protocol analysis (n = 41 pimecrolimus; 46 vehicle) indicated a significant difference between groups (mean change 3.9 pimecrolimus vs 3.2 vehicle; P = .0156; CI [0.129, 1.197]). The superiority of pimecrolimus was observed as early as week 2 (intent-to-treat: P = .0062; CI [0.132, 0.777]; per protocol: P = .0012; CI [0.410, 1.593]). No drug-related serious adverse events occurred. The most frequent drug-related adverse events were local, mild, and transient (pimecrolimus = 26%; vehicle = 12%). Generalizability is limited by the elderly male study population. This study suggests that pimecrolimus cream 1% is an effective and well-tolerated treatment for moderate to severe facial seborrheic dermatitis.


Comparative proteomic analysis of generative and sperm cells reveals molecular characteristics associated with sperm development and function specialization.
In flowering plants, two sperm cells (SCs) are generated from a generative cell (GC) in the developing pollen grain or growing pollen tube and are then delivered to the embryo sac to initiate double fertilization. SC development and function specialization involve the strict control of the protein (gene) expression program and coordination of diverse cellular processes. However, because methods for collecting a large amount of highly purified GCs and SCs for proteomic and transcriptomic studies from a plant are not available, molecular information about the program and the interconnections is lacking. Here, we describe a method for obtaining a large quantity of highly purified GCs and SCs from just-germinated lily pollen grains and growing pollen tubes for proteomic analysis. Our observation showed that SCs had less condensed chromatin and more vacuole-like structures than GCs and that mature SCs were arrested at the G2 phase. Comparison of SC and GC proteomes revealed 101 proteins differentially expressed in the two proteomes. These proteins are involved in diverse cellular and metabolic processes, with preferential involvement in metabolism, the cell cycle, signaling, the ubiquitin/proteasome pathway, and chromatin remodeling. Impressively, almost all proteins in SCF complex-mediated proteolysis and the cell cycle were up-regulated in SCs, whereas those in chromatin remodeling and stress response were down-regulated. Our data also reveal the coordination of SCF complex-mediated proteolysis, cell cycle progression, and DNA repair in SC development and function specialization. This study revealed for the first time a difference in protein profiles between GCs and SCs.


Assessment of mold contamination and physicochemical properties of crude palm oil sold in Jos, Nigeria.
Due to increasing reports on poor-quality palm oil in the market, there has been a continual decrease in demand and revenue cum product rejection of palm oil sold in Jos, Nigeria. Hence, the significance of this work aims to moderate the microbial and physical qualities of crude palm oil sold in different major markets in order to increase revenue through quality control and quality assurance protocols. The intent is to create awareness among government monitoring agencies, buyers (exporters and importers), and to promote standard processing procedures among manufacturers. The study was carried out to ascertain the levels of mold contamination and physicochemical properties of crude palm oil sold in Jos and its environ. A total of 90 samples were collected in sterile containers. Molds were isolated and identified using standard identification procedures. The physicochemical properties: free fatty acids (FFA), peroxide value (PV), iodine value (IV), moisture content (MC), and impurity level were determined. The assessment of mold isolated from the study sites recognized some life-threatening genera capable of producing carcinogenic toxins. Candida sp. (51%) had the highest percentage of occurrence followed by Aspergillus sp. (45%) while Fusarium sp. (3%) was the least occurring mold. The mean mold count for all the palm oil samples ranges from 3.18 × 104 (cfu/mL)-4.56 × 104 (cfu/mL). Physicochemical findings showed that the average FFA (3.43-6.88%), PV (6.96-13.63 mEq/kg), IV (39.7-67.5 wijs), and MC (0.44-0.72%) values were within the SON acceptable limit, except the impurity level (0.28-0.44%) which was higher than the acceptable SON range in all the sites. There was a significant difference (P ≤ 0.05) in the mold and physicochemical properties of crude palm oil in almost all the samples analyzed when compared to both local and international permissible limits of the tested parameters recommended for edible palm oil. There was compliance between the permissible limits (local and international) of the physicochemical values of the parameters we tested for in the edible palm oil except the impurity level, while the mold count did not meet (was higher) with the required permissible limit of the SON mold count.


The effect of lymphatic oedema on the uptake of colloids to the lymph nodes.
The majority of pathological conditions of the lymphatic system can result in some degree of lymphoedema, which in turn causes a reduced rate of lymph flow. In some cases, such as when nodes are invaded by tumour metastases, blockage of the lymphatic vessels may occur. In order to investigate the effect of such pathology on nanosphere uptake in regional lymph nodes, the fate of model polystyrene nanospheres, surface modified with block co-polymers of the poloxamine series, was determined following subcutaneous administration in a rat model with lymphoedema (induced by the administration of lambda-carrageenan). A drastic reduction of injection site drainage and lymph node uptake of nanospheres was observed in the inflammation model compared to control animals. The observations suggest that biodegradable nanospheres based on these will be suitable for the detection of oedema in the lymphatic system.


Limited niche availability suppresses murine intrathymic dendritic-cell development from noncommitted progenitors.
The origins of dendritic cells (DCs) and other myeloid cells in the thymus have remained controversial. In this study, we assessed developmental relationships between thymic dendritic cells and thymocytes, employing retrovirus-based cellular barcoding and reporter mice, as well as intrathymic transfers coupled with DC depletion. We demonstrated that a subset of early T-lineage progenitors expressed CX3CR1, a bona fide marker for DC progenitors. However, intrathymic transfers into nonmanipulated mice, as well as retroviral barcoding, indicated that thymic dendritic cells and thymocytes were largely of distinct developmental origin. In contrast, intrathymic transfers after in vivo depletion of DCs resulted in intrathymic development of non-T-lineage cells. In conclusion, our data support a model in which the adoption of T-lineage fate by noncommitted progenitors at steady state is enforced by signals from the thymic microenvironment unless niches promoting alternative lineage fates become available.


Multidisciplinary Management of Ataxia Telangiectasia: Current Perspectives.
Ataxia telangiectasia (A-T) is a rare autosomal recessive disease caused by mutations in the ataxia telangiectasia mutated (ATM) gene. In the absence of a family history, the diagnosis of A-T is usually not made until the child is older and symptomatic. Classic A-T is characterized by a constellation of clinical symptoms including progressive ataxia, oculocutaneous telangiectasias and sinopulmonary disease and is usually associated with absence of ATM protein. Other laboratory features associated with A-T include elevated serum levels of alpha-fetoprotein (AFP) and increased chromosomal breakage with in vitro exposure to ionizing radiation. Sinopulmonary symptoms can occur to varying degrees across the lifespan. Some children will also have hypogammaglobulinemia and impaired antibody responses requiring supplemental gamma globulin. People with hypomorphic ATM mutations are often considered to have mild A-T with onset of ataxia and neurological progression occurring later in life with less impairment of the immune system. The risk of malignancy, however, is significantly increased in people with either classic or mild A-T. While hematological malignancies are most common in the first two decades of life, solid organ malignancies become increasingly common during young adulthood. Deterioration of neurologic function with age is associated with dysphagia with aspiration, growth faltering, loss of ambulation and decline in pulmonary function, morbidities that contribute to shortened life expectancy and decreased quality of life. Premature death is often due to malignancies or chronic respiratory insufficiency. A-T is currently managed with supportive care and symptomatic treatment. Current clinical trials, however, represent progress and hope towards disease-modifying therapies for A-T.


Decreased catalytic activity and expression of protein kinase C isozymes in teenage suicide victims: a postmortem brain study.
Teenage suicide is a major public health concern. Although there is some understanding of the psychosocial factors associated with teenage suicide, little is known about the neurobiologic factors of teenage suicide. Protein kinase C (PKC) is a critical phosphorylating enzyme in the phosphoinositide signaling pathway (which is involved in many physiologic functions in the brain and has been implicated in the pathogenesis of mood disorders) and is also a target for the therapeutic action of mood-stabilizing drugs. To examine whether the pathogenesis of teenage suicide is associated with changes in PKC. Postmortem brain study. Seventeen teenage suicide victims and 17 nonpsychiatric control subjects. Catalytic activity of PKC and protein and messenger RNA levels of various PKC isozymes, such as PKC alpha, beta, and gamma, were determined in the prefrontal cortex and hippocampus of both groups. Protein kinase C activity was statistically significantly decreased in membrane and cytosol fractions of the prefrontal cortex and hippocampus of teenage suicide victims compared with control subjects. Statistically significant decreases in protein levels of PKC alpha, beta I, beta II, and gamma isozymes were also observed in both of these fractions. These decreases were associated with decreases in levels of their respective messenger RNAs. Because many physiologic functions are mediated through phosphorylation by PKC and because PKC is a target for the therapeutic action of psychoactive drugs, our findings indicate that the pathogenesis of teenage suicide may be associated with abnormalities in PKC and that PKC may be a target for therapeutic intervention in patients with suicidal behavior.


Laboratory Automation in Microbiology: Impact on Turnaround Time of Microbiological Samples in COVID Time.
Laboratory Automation (LA) is an innovative technology that is currently available for microbiology laboratories. LA can be a game changer by revolutionizing laboratory workflows through efficiency improvement and is also effective in the organization and standardization of procedures, enabling staff requalification. It can provide an important return on investment (time spent redefining the workflow as well as direct costs of instrumentation) in the medium to long term. Here, we present our experience with the WASPLab® system introduced in our lab during the COVID-19 pandemic. We evaluated the impact due to the system by comparing the TAT recorded on our samples before, during, and after LA introduction (from 2019 to 2021). We focused our attention on blood cultures (BCs) and biological fluid samples (BLs). TAT recorded over time showed a significant decrease: from 97 h to 53.5 h (Δ43.5 h) for BCs and from 73 h to 58 h (Δ20 h) for BLs. Despite the introduction of the WASPLab® system, we have not been able to reduce the number of technical personnel units dedicated to the microbiology lab, but WASPLab® has allowed us to direct some of the staff resources toward other laboratory activities, including those required by the pandemic. LA can significantly enhance laboratory performance and, due to the significant reduction in reporting time, can have an effective impact on clinical choices and therefore on patient outcomes. Therefore, the initial costs of LA adoption must be considered worthwhile.


Job Satisfaction and Turnover Intention Among People With Disabilities Working in Special Employment Centers: The Moderation Effect of Organizational Commitment.
With the goal of contributing to the growth of research on people with disabilities in employment, in particular in relation to their job satisfaction (JS), organizational commitment (OC), and turnover intention (TI), this study explores the effect of JS on TI among employees with disabilities and the moderation effect of OC and its four dimensions on this main relationship. A total of 245 Special Employment Center (SEC) employees in Spain answered a questionnaire. To analyze the results, a descriptive analysis with bivariate correlations across the variables was performed, and the moderation model was tested subsequently using macro PROCESS for SPSS by Hayes. For the significant effects, a pick-a-point approximation was used to interpret the results. The results show that OC and its dimensions have no significant effect on the direct relationship. However, some components of JS, such as the relationship with co-workers and with supervisors, play a significant role in the relationship with TI when moderated by affective and value commitment. Our results show that it is important that human resources departments create conditions favoring a work environment with positive interpersonal relationships between employees and managers in order to minimize TI at SECs.


Increases in number and size of osteoclasts in response to calcium or phosphorus deficiency in the rat.
In this study, the two determinants of the rate of osteoclastic bone resorption, cell number and cell activity, were evaluated to characterize the response to two chronic resorptive stimuli, a calcium-deficient diet and a phosphorus-deficient diet, fed to rats for 8 days. The number of osteoclast nuclei was determined directly and the resorptive activity of osteoclasts indirectly from cell size. In our sampling site in the tibial diaphysis, there were marked increases in osteoclast layer width (i.e., cell size) and in the number of osteoclasts and osteoclast nuclei in both test groups. Also, in both test groups the increase in the number of osteoclast nuclei was greater than the increase in osteoclast width, emphasizing the importance of osteoclast stem cell activity in the resorptive response. When the data on the control and test animals were pooled, there was a strong correlation between the width of the osteoclast layer and the number of osteoclast nuclei (r = 0.87, P less than 0.001). Thus, these two resoprtive stimuli produced proportional changes in osteoclast cell number and probably activity, raising the possibility that the resorptive response is sterotyped. These results also suggest the existence of a control mechanism which coupled the proliferative activity of osteoclast stem cells and the resorptive activity of osteoclasts. Despite the fact that the rats were subjected to chronic resorptive stimuli, forming surface and the total number of osteoblasts were also increased. Moreover, in data pooled from control and test rats, there was a strong correlation between the number of osteoclast nuclei and the number of osteoblasts (r = 0.92, P less than 0.001). This implies that chronic resorptive stimuli directly or indirectly stimulate osteogenic stem cells to increase production of osteoblasts as well as osteoclasts. The observed coupling of differentiated cell number probably contributes to the established phenomenon of coupling between the rates of bone accretion and resorption.


A misclassification model for the non-disjunction fraction in meiosis I.
In this paper we introduce a misclassification model for the meiosis I non-disjunction fraction in numerical chromosomal anomalies named trisomies. We obtain posteriors, and their moments, for the probability that a non-disjunction occurs in the first division of meiosis and for the misclassification errors. We also extend previous works by providing the exact posterior, and its moments, for the probability that a non-disjunction occurs in the first division of meiosis assuming the model proposed in the literature which does not consider that data are subject to misclassification. We perform Monte Carlo studies in order to compare Bayes estimates obtained by using both models. An application to Down Syndrome data is also presented.


[Free Radical Processes Play an Important Role in the Etiology and Pathogenesis of Atherosclerosis and Diabetes].
In this review, we present with exhaustive completeness the data of foreign studies and results of many years of own research, evidencing of the important role of free radical processes in the etiology and pathogenesis of atherosclerosis. The review contains the rationale for the concept that oxidative stress in atherosclerosis is even more developed during disorders of carbohydrate metabolism, contributing to the emergence of carbonyl stress in diabetes. We hypothesize that there exists a single common molecular mechanism of primary preatherogenic damages of vessel walls in atherosclerosis and diabetes, which enhances the formation of carbonyl-modified low-density lipoproteins to be accumulated in the foam cells. Possible reasons for failure of "antioxidant therapy" of atherosclerosis are discussed.


Biochemical effects of Solidago virgaurea extract on experimental cardiotoxicity.
Cardiovascular diseases (CVDs) are the major health problem of advanced as well as developing countries of the world. The aim of the present study was to investigate the protective effect of the Solidago virgaurea extract on isoproterenol-induced cardiotoxicity in rats. The subcutaneous injection of isoproterenol (30 mg/kg) into rats twice at an interval of 24 h, for two consecutive days, led to a significant increase in serum lactate dehydrogenase, creatine phosphokinase, alanine transaminase, aspartate transaminase, and angiotensin-converting enzyme activities, total cholesterol, triglycerides, free serum fatty acid, cardiac tissue malondialdehyde (MDA), and nitric oxide levels and a significant decrease in levels of glutathione and superoxide dismutase in cardiac tissue as compared to the normal control group (P < 0.05). Pretreatment with S. virgaurea extract for 5 weeks at a dose of 250 mg/kg followed by isoproterenol injection significantly prevented the observed alterations. Captopril (50 mg/kg/day, given orally), an inhibitor of angiotensin-converting enzyme used as a standard cardioprotective drug, was used as a positive control in this study. The data of the present study suggest that S. virgaurea extract exerts its protective effect by decreasing MDA level and increasing the antioxidant status in isoproterenol-treated rats. The study emphasizes the beneficial action of S. virgaurea extract as a cardioprotective agent.


How to Conduct Clinical Trials in Children: A Tutorial.
Despite a growing interest in, and commitment to, implementing pediatric clinical trials, approximately one in every five trials in children fails because of inappropriate study design, suboptimal experiment planning, or inadequate participant enrollment. This tutorial, presented from the perspectives of seasoned pediatric investigators, an experienced research coordinator, and an established pediatric clinical trials network, is designed to provide practical guidance for successfully implementing pediatric clinical trials at an academic center or another comparable institution.


Anxiety-like behaviour assessments of adolescent rats after repeated maternal separation during early life.
Maternal separation (MS) plays a central role in developing physiology and psychology during the individual ontogeny process. MS is used to research the neurobiological mechanisms of mental disorders and early life stress. In this study, we investigated the effects of repeated MS and early handling (EH) on locomotor activity in an open-field test, a light-dark box test and an elevated plus-maze test of adolescent rats. The results showed that MS reduced locomotor activities in the open-field test, and increased anxiety-like behaviours in the light-dark box test and the elevated plus-maze test in adolescent rats. These tests indicated that early life stress caused by MS might induce anxiety-like behaviours during adolescence. However, compared with the control group, both the MS and EH groups showed conflicting anxiety levels. The results also suggested that females were more prone to showing anxiety-like behaviour compared with males when suffering from high-intensity stimulation. However, because of the low anxiety level associated with EH, the sex difference in behaviour was not significant. The present study provides novel insights into the effects of MS and EH on behaviour, which shows unique anxiety levels different in adolescent male and female rats.


New bioactive peptide reduces the toxicity of chemotherapy drugs and increases drug sensitivity.
Anticancer bioactive peptide (ACBP) is extracted from normal goat spleens and exhibits antitumor activity alone and in combination with low cisplatin doses to achieve antitumor efficacy similar to higher cisplatin doses via sustained medication modes. In the present study, we investigated whether elevated levels of induced or normal ACBP in MKN‑45 gastric cancer (GC) cells may reduce their toxicity to oxaliplatin (L‑OHP) in a dose‑dependent manner. The growth inhibition rate (IR), morphological changes and gene expression were examined in MKN‑45 GC cells. Compared with normal ACBP, induced ACBP alone significantly enhanced the anticancer activity of L‑OHP‑mediated apoptosis and reduced the amount and side‑effects of L‑OHP (P<0.05). The inhibition of cancer cell growth at high concentrations of induced ACBP and L‑OHP was significantly more effective than at low concentrations. In addition, for the first time, we examined the potential of a combination of induced ACBP and L‑OHP to increase L‑OHP sensitivity in human gastric carcinoma xenograft tumors. Nude mice were implanted with human gastric carcinoma MKN‑45 cells and treated with an intraperitoneal injection of 0.5 ml of normal saline, 30 µg/ml ACBP, 20 µg/ml L‑OHP or 30 µg/ml ACBP + 20 µg/ml L‑OHP [combination of anticancer bioactive peptide and oxaliplatin (A+L)] via the tail vein twice a week. In vivo short‑term intermittent use of induced ACBP alone significantly inhibited MKN‑45 tumor growth. The combination of induced ACBP and L‑OHP also significantly improved the quality of life of the nude mice and reduced the toxicity of L‑OHP. Based on flow cytometry and gene expression analyses, A+L significantly increased the proportion of cells in the G2/M phase (P<0.05) relative to ACBP or L‑OHP alone, and short‑term intraperitoneal injection of ACBP increased the sensitizing effect of L‑OHP. Collectively, these results suggest that high levels of induced ACBP in combination with L‑OHP via a short‑term intermittent medication mode could be a useful clinical therapeutic strategy for GC.


Juvenile hormone acid methyltransferase: a key regulatory enzyme for insect metamorphosis.
Juvenile hormone (JH) acid methyltransferase (JHAMT) is an enzyme that converts JH acids or inactive precursors of JHs to active JHs at the final step of JH biosynthesis pathway in insects. By fluorescent mRNA differential display, we have cloned a cDNA encoding JHAMT from the corpora allata (CA) of the silkworm, Bombyx mori (BmJHAMT). The BmJHAMT cDNA encodes an ORF of 278 aa with a calculated molecular mass of 32,544 Da. The predicted amino acid sequence contains a conserved S-adenosyl-l-methionine (SAM) binding motif found in the family of SAM-dependent methyltransferases. Purified N-terminal 6xHis-tagged recombinant BmJHAMT protein expressed in Escherichia coli catalyzed conversion of farnesoic acid and JH acids I, II, and III to their cognate methyl esters in the presence of SAM, confirming that this cDNA encodes a functional JHAMT. Putative orthologs, DmJHAMT and AgJHAMT, were identified from the genome sequence of the fruit fly Drosophila melanogaster, and a malaria vector, Anopheles gambiae, respectively. Northern blot and quantitative RT-PCR analyses revealed that the BmJHAMT gene was expressed specifically in the CA throughout the third and fourth instar. At the beginning of the last (fifth) instar, the expression level of BmJHAMT declined rapidly and became undetectable by day 4 and remained so until pupation. Correlation of the BmJHAMT gene expression and the JH biosynthetic activity in the CA suggests that the transcriptional suppression of the BmJHAMT gene is crucial for the termination of JH biosynthesis in the CA, which is a prerequisite for the initiation of metamorphosis.


Potential salivary biomarkers and their genetic effects in a pilot study of adolescent boys with externalizing problems.
Beside the well-known stress response marker cortisol, salivary alpha-amylase is receiving increasing attention. Numerous studies have investigated the potential biomarker properties of cortisol mirroring abnormal hypothalamic-pituitary-adrenal axis activity in connection to both internalizing and externalizing behavior problems. The other major physiological system involved in stress reactivity, the sympathetic nervous system activity can be also measured by the surrogate marker of salivary alpha-amylase. Most of the studies applied a stressful situation to obtain inter-individual differences in stress-reactivity, although differences in the baseline level of cortisol have been also shown in relation to externalizing problems. To test the relevance of another (easier) biomarker, we selected to study baseline circadian salivary cortisol and alpha-amylase levels among adolescent boys with externalizing problems. Saliva samples were collected at 3 time-points (morning, noon, evening) during 3 consecutive days from 37 inpatient boys (mean age 12.4±1.0). Cortisol and alpha-amylase levels were measured by enzyme-linked immunosorbent and kinetic enzyme assays, respectively. Genetic variants in the hypothalamic-pituitary-adrenal axis and the norepinephrine transporter or catecholamine metabolizing enzymes were tested for potential moderating effects at these salivary biomarkers. Saliva cortisol showed the classical diurnal fluctuation in boys with externalizing problems (possibly from a lower morning level), but it was not modified by the presence of either conduct, oppositional defiant or attention-deficit/hyperactivity disorder. The diurnal fluctuation of the salivary alpha-amylase levels was also typical, but the presence of conduct disorder was associated with significantly lower alpha-amylase activity (p=0.024) among boys with externalizing problems. The catechol-O-methyltransferase Val158Met (rs4680) polymorphism had an additional effect on salivary alpha-amylase: boys with homozygote genotypes had lower alpha-amylase activity at all 3 time-points compared to Val/Met heterozygotes (p=0.045). Our preliminary data suggest that salivary alpha-amylase might be used to further characterize subgroups within externalizing problems, however, this biomarker might be modified by certain genetic polymorphisms.


A one-dimensional coordination polymer constructed from isatine-3-oximate and sodium.
The reaction of hydroxyl-amine hydro-chloride with isatin in ethanol, catalysed with HCl and neutralized with Na(2)CO(3), yielded the one-dimensional coordination polymer, catena-poly[[[aqua-sodium]-di-μ-aqua-[aqua-sodium]-bis-(μ-2-oxoindoline-2,3-dione 3-oximato)] tetra-kis-(oxoindoline-2,3-dione 3-oxime)], {[Na(C(8)H(5)N(2)O(2))(H(2)O)(2)]·2C(8)H(6)N(2)O(2)}(n). The Na(I) atom has a six-coordinate distorted-octa-hedral environment. Isatine-3-oximate O atoms and water mol-ecules bridge adjacent Na atoms, forming a one-dimensional polymeric structure parallel to [100]. Each isatine-3-oxime dimerizes through N-H⋯O interactions and in addition each oxime is linked to a coordination polymer. Thus, coordination polymers are linked by O-H⋯O and O-H⋯N interactions from isatine-3-oxime dimers, building a two-dimensional network parallel to [110].


Purification of a maize dehydrin.
A maize dehydrin with an apparent molecular weight of 20 kDa was purified from whole kernels of maize inbred line G50. Kernels were ground in a seed mill, stirred overnight in extraction buffer, and centrifuged to extract soluble proteins. The sample was heated to 89 degrees C and centrifuged to remove heat-insoluble proteins. The remaining soluble proteins were fractionated in a three-step chromatographic process. Following cation exchange, hydrophobic interaction, and gel filtration chromatography, pure dehydrin samples were obtained.


Susceptibility of intact germinating Arabidopsis thaliana to human fungal pathogens Cryptococcus neoformans and C. gattii.
The fungus Cryptococcus contributes a large global burden of infectious death in both HIV-infected and healthy individuals. As Cryptococcus is an opportunistic pathogen, much of the evolutionary pressure shaping virulence occurs in environments in contact with plants and soil. The present studies investigated inoculation of intact seeds of the common weed Arabidopsis thaliana with fungal cells over a 21-day period. C. gattii was the more virulent plant pathogen, resulting in disrupted germination as well as increased stem lodging, fungal burden, and plant tissue colocalization. C. neoformans was a less virulent plant pathogen but exhibited prolonged tissue residence within the cuticle and vascular spaces. Arabidopsis mutants of the PRN1 gene, which is involved in abiotic and biotic signaling affecting phenylalanine-derived flavonoids, showed altered susceptibility to cryptoccocal infections, suggesting roles for this pathway in cryptococcal defense. The fungal virulence factor laccase was also implicated in plant pathogenesis, as a cryptococcal lac1Δ strain was less virulent than wild-type fungi and was unable to colonize seedlings. In conclusion, these studies expand knowledge concerning the ecological niche of Cryptococcus by demonstrating the pathogenic capacity of the anamorphic form of cryptococcal cells against healthy seedlings under physiologically relevant conditions. In addition, an important role of laccase in plant as well as human virulence may suggest mechanisms for laccase retention and optimization during evolution of this fungal pathogen.


Association of Combined Focal 22q11.22 Deletion and IKZF1 Alterations With Outcomes in Childhood Acute Lymphoblastic Leukemia.
Alterations in the IKZF1 gene drive B-cell acute lymphoblastic leukemia (B-ALL) but are not routinely used to stratify patients by risk because of inconsistent associations with outcomes. We describe a novel deletion in 22q11.22 that was consistently associated with very poor outcomes in patients with B-ALL with IKZF1 alterations. To determine whether focal deletions within the λ variable chain region in chromosome 22q11.22 were associated with patients with B-ALL with IKZF1 alterations with the highest risk of relapse and/or death. This cohort study included 1310 primarily high-risk pediatric patients with B-ALL who were taken from 6 independent clinical cohorts, consisting of 3 multicenter cohorts (AALL0232 [2004-2011], P9906 [2000-2003], and patients with Down syndrome who were pooled from national and international studies) and 3 single-institution cohorts (University of Utah [Salt Lake City], Children's Hospital of Philadelphia [Philadelphia, Pennsylvania], and St. Jude Children's Hospital [Memphis, Tennessee]). Data analysis began in 2011 using patients from the older studies first, and data analysis concluded in 2021. Focal 22q11.22 deletions. Event-free and overall survival was investigated. The hypothesis that 22q11.22 deletions stratified the prognostic effect of IKZF1 alterations was formulated while investigating nearby deletions in VPREB1 in 2 initial cohorts (n = 270). Four additional cohorts were then obtained to further study this association (n = 1040). This study of 1310 patients with B-ALL (717 male [56.1%] and 562 female patients [43.9%]) found that focal 22q11.22 deletions are frequent (518 of 1310 [39.5%]) in B-ALL and inconsistent with physiologic V(D)J recombination. A total of 299 of 1310 patients with B-ALL had IKZF1 alterations. Among patients with IKZF1 alterations, more than half shared concomitant focal 22q11.22 deletions (159 of 299 [53.0%]). Patients with combined IKZF1 alterations and 22q11.22 deletions had worse outcomes compared with patients with IKZF1 alterations and wild-type 22q11.22 alleles in every cohort examined (combined cohorts: 5-year event-free survival rates, 43.3% vs 68.5%; hazard ratio [HR], 2.18; 95% CI, 1.54-3.07; P < .001; 5-year overall survival rates, 66.9% vs 83.9%; HR, 2.05; 95% CI, 1.32-3.21; P = .001). While 22q11.22 deletions were not prognostic in patients with wild-type IKZF1 , concomitant 22q11.22 deletions in patients with IKZF1 alterations stratified outcomes across additional risk groups, including patients who met the IKZF1plus criteria, and maintained independent significance in multivariate analysis for event-free survival (HR, 2.05; 95% CI, 1.27-3.29; P = .003) and overall survival (HR, 1.83; 95% CI, 1.01-3.34; P = .05). This cohort study suggests that 22q11.22 deletions identify patients with B-ALL and IKZF1 alterations who have very poor outcomes and may offer a new genetic biomarker to further refine B-ALL risk stratification and treatment strategies.


Reconstruction of Highly Proliferative Auto-Tissue-Engineered Lamellar Cornea Enhanced by Embryonic Stem Cell.
To increase the epithelial proliferation of an auto-tissue-engineered lamellar cornea, 3.0 × 10(6) corneal epithelial cells (CECs) were combined with 3 × 10(5) mouse embryonic stem cells (ESCs) pretransfected with the HSV-tk gene (CECs + ESCs-TK group), and 3.3×10(6) corneal epithelial cells (CECs group) were seeded between the acellular porcine corneal stroma and the amniotic membrane using the centrifugal cell seeding method. After 4 days of perfusion culture (treatment with ganciclovir starting on day 2), a thicker corneal epithelium (four to five layers) formed in the CECs + ESCs-TK group compared with that observed in the CECs group (two to three layers). More stem/progenitor cell (K3 -, p63+, ABCG2+, and integrin-β1+) and proliferation phenotypes (Ki67+) were measured in the CECs + ESCs-TK group compared with the CECs group using immunofluorescence staining, real-time quantitative reverse transcription polymerase chain reaction, and flow cytometry. Consistent with these findings, the colony-forming efficiency and cellular doubling time were significantly different between the CECs + ESCs-TK group (16.18% ± 3.98%, 28.45 ± 2.03 h) and CECs group (11.96% ± 2.60%, 36.3 ± 1.15 h). In a rabbit lamellar transplantation model, the CECs + ESCs-TK group had better epithelial barrier functions and wound healing abilities compared with the CECs group. Furthermore, ESCs-TK could be completely and safely removed by ganciclovir. Thus, the ESCS-TK coculture system could serve as a potential strategy for corneal tissue engineering.


Comparison of smoking cessation between education groups: findings from 2 US National Surveys over 2 decades.
We examined smoking cessation rate by education and determined how much of the difference can be attributed to the rate of quit attempts and how much to the success of these attempts. We analyzed data from the National Health Interview Survey (NHIS, 1991-2010) and the Tobacco Use Supplement to the Current Population Survey (TUS-CPS, 1992-2011). Smokers (≥ 25 years) were divided into lower- and higher-education groups (≤ 12 years and >12 years). A significant difference in cessation rate between the lower- and the higher-education groups persisted over the last 2 decades. On average, the annual cessation rate for the former was about two thirds that of the latter (3.5% vs 5.2%; P<.001, for both NHIS and TUS-CPS). About half the difference in cessation rate can be attributed to the difference in quit attempt rate and half to the difference in success rate. Smokers in the lower-education group have consistently lagged behind their higher-education counterparts in quitting. In addition to the usual concern about improving their success in quitting, tobacco control programs need to find ways to increase quit attempts in this group.


Comparison of CT duodeno-cholangiopancreatography to ERCP for assessing biliary obstruction.
The authors have developed a CT protocol, CT duodeno-cholangiopancreatography (CDCP), which is performed during a single contrast-enhanced phase, proceeding cranially, allowing enhancement of the pancreas during its parenchymal phase followed by enhancement of the liver during its portal-venous phase. This retrospective pilot study evaluates CDCP compared with endoscopic retrograde cholangiopancreatography (ERCP) as a diagnostic tool for assessing the cause and level of biliary obstruction. Forty-one patients with jaundice underwent CDCP and ERCP between October 2002 and May 2004. Pathologic confirmation was obtained in 31 of the 41 (76%) patients. The sensitivity, specificity, and kappa values of CDCP and ERCP compared with pathology were calculated for tumors and stones. Pathology-proven cases included 7 cases of stones, 23 tumors, and 1 other cause of obstruction. The overall level of agreement of diagnoses between CDCP and pathology was 29 of 31 (93.5%); that between CDCP and ERCP was 36 of 41 (88%). Comparing CDCP to pathology for tumors, the sensitivity was 100%, the specificity was 89%, and the kappa was 0.92 (95% CI 0.76-1.0). For stone detection, CDCP had a sensitivity of 86%, a specificity of 100%, and a kappa value of 0.90 (95% CI 0.72-1.0). For level of obstruction of the common bile duct, comparing CDCP to ERCP, observations agreed in 31 of the 36 (86%) cases; for the pancreatic duct, observations agreed in 24 of the 25 (96%) cases. CDCP is a noninvasive diagnostic tool that can be used to assess the cause and level of obstruction. A blinded prospective study would be valuable to further assess the merits of CDCP.


The 2020 update on anaphylaxis in paediatric population.
The objective of the review is to present recent updates on anaphylaxis in paediatric population worldwide. The article summarizes the results of epidemiological studies, diagnostic methods and treatments. We present a new WAO definition of anaphylaxis (2019), which broader criteria excluding dermal symptoms should facilitate faster life-saving adrenaline use. Adrenaline remains the best treatment to manage severe symptoms and to prevent biphasic reactions. There is ongoing effort to increase adrenaline use, such as modified autoinjectors, individual training, and diversified dosing. There are five independent risk factors of lethal anaphylaxis in children, including history of asthma, almost immediate onset of symptoms, unwell appearance, tachycardia and hypotension. We also report improvements in diagnostics, like component-resolved diagnostics, and novel therapies stimulating immunotolerance. We signal the development of ICD-11 with updated coding of anaphylaxis, which corresponds better to clinical observations.


Synthetic Glycosphingolipids for Live-Cell Labeling.
Glycosphingolipids are an important component of cell membranes that are involved in many biological processes. Fluorescently labeled glycosphingolipids are frequently used to gain insight into their localization. However, the attachment of a fluorophore to the glycan part or-more commonly-to the lipid part of glycosphingolipids is known to alter the biophysical properties and can perturb the biological function of the probe. Presented here is the synthesis of novel glycosphingolipid probes with mono- and disaccharide head groups and ceramide moieties containing fatty acids of varying chain length (C4 to C20). These glycosphingolipids bear an azide or an alkyne group as chemical reporter to which a fluorophore can be attached through a bioorthogonal ligation reaction. The fluorescent tag and any linker connected to it can be chosen in a flexible manner. We demonstrate the suitability of the probes by selective visualization of the plasma membrane of living cells by confocal microscopy techniques. Whereas the derivatives with the shorter fatty acids can be directly applied to HEK 293T cells, the hydrophobic glycosphingolipids with longer fatty acids can be delivered to cells using fusogenic liposomes.


Winged fruits and associated leaves of Shorea (Dipterocarpaceae) from the Late Eocene of South China and their phytogeographic and paleoclimatic implications.
Dipterocarps are the representative component of tropical rain forests in Southeast Asia and hold important economic and ecological significance, but their origin and migration are controversial. Information on dipterocarpaceous fossils, particularly the more convincing reproductive structures, not only can improve the phylogenetic and phytogeographic studies of this family, but also provide important information for reconstructing paleoclimate. • Morphologically preserved winged fruits and associated leaves were collected from the Late Eocene Huangniuling Formation, Maoming Basin, South China. We determined their taxonomic positions based on comparative morphology with similar extant and fossil specimens and discuss their phytogeographic and paleoclimatic implications by consulting the distribution and habitat of fossil and modern populations. • The Late Eocene winged fruits are attributed to Shorea Roxburgh ex Gaertner (Dipterocarpaceae) as Shorea maomingensis sp. nov. The associated leaves are recognized as Shorea sp. based on leaf architecture, and they are likely to be conspecific with the winged fruits. • The discovery of dipterocarps indicates that they had arrived in tropical and humid South China by the Late Eocene. Dipterocarps including Shorea exhibit a wide range of physiological tolerance to climate; palynological analysis suggests an increase in aridity and seasonality in the Maoming Basin from the Late Eocene. Dipterocarps became adapted to this seasonal climate from the Late Eocene to Early Miocene, expanded northward in the climatic optimum of the Middle Miocene, and declined and gradually disappeared from the southeastern part of the continent from the Late Miocene.


Gastrointestinal stromal tumours: evaluation of biological and clinical current opinions.
The discovery of c-kit gene mutations and the positivity of its transcription products in gastrointestinal stromal tumours (GISTs) suggest a possible origin from Cajal interstitial cells. The study population consisted of 12 patients with GIST, with a mean age of 67.6 years. Preoperative biopsy was performed in 4 cases and in only 1 case did it prove correct. Mesenchymal tumours were regarded as myogenic when they were positive for desmin, and as neurogenic when they were positive for S-100 protein or specific neural enolase (SNE). Seven out of 12 patients underwent simple tumour excision, while in 2 cases ileal resection was performed; gastric resection, total gastrectomy with D2 lymphadenectomy and left colectomy were carried out in one case each. There were two deaths, both unrelated to the primitive diagnosis. Immunohistochemical studies were positive for CD34 in 58% of the cases, and for CD117 in 83%. The mitotic count was higher than 5/10 HPF in 3 cases. The mean survival was 57 months. The overall survival rate was 66%. We found that good tumour differentiation, small size and a low mitotic index correlate with benign behaviour and a better prognosis. Positivity for CD117, evaluated in all malignant lesions, was slightly lower (83% vs 89%) as compared to the data reported in the literature. Tumours with a high mitotic index (> 5 mitoses/10 HPF) and measuring more than 5 cm in diameter are to be considered malignant.


Effect of positive charges in the structural interaction of crabrolin isoforms with lipopolysaccharide.
Antimicrobial peptides (AMPs) appear as chemical compounds of increasing interest for their role in killing bacteria and, more recently, for their ability to bind endotoxin (lipopolysaccharide, LPS) that is released during bacterial infection and that may lead to septic shock. This dual role in the mechanism of action can further be enhanced in a synergistic way when two or more AMPs are combined together. Not all AMPs are able to bind LPS, suggesting that several modes of binding to the bacterial surface may exist. Here we analyze a natural AMP, crabrolin, and two mutated forms, one with increased positive charge (Crabrolin Plus) and the other with null charge (Crabrolin Minus), and compare their binding abilities to LPS. While Crabrolin WT as well Crabrolin Minus do not show binding to LPS, the mutated Crabrolin Plus exhibits binding and forms a well defined structure in the presence of LPS. The results strengthen the importance of positive charges for the binding to LPS and suggest the mutated form with increased positive charge as a promising candidate for antimicrobial and antiseptic activity.


Clinical and serological evaluation of Leptospirosis in Puducherry, India.
Leptospirosis is a zoonotic disease with a worldwide distribution. There is a paucity of available data about prevalence of this disease in Pondicherry. Our aim was to investigate the seropositivity rate of leptospirosis in suspected cases and also to identify the predominant serogroups present by performing Microscopic Agglutination Test (MAT). The other aim of this study was to compare the results of a commercially available IgM ELISA with that of MAT. A total of 110 blood samples from patients suspected of leptospirosis were sent for diagnosis. These samples were subjected to IgM ELISA and the microscopic agglutination test (MAT). MAT was done using a panel of 12 Leptospira serovars. MAT analysis of the 110 samples showed 40 (36%) to be positive. Antibodies were predominantly seen against serogroup Leptospira Icterohemorrhagiae (27%), followed by Pomona (17%), and Pyrogenes (12%). IgM ELISA done on these samples showed a positivity of 37% compared to MAT. This study reveals that the MAT test can be standardized in a diagnostic laboratory and used in conjunction with an IgM ELISA.


The Nithsdale schizophrenia survey. IV. Social adjustment by self-report.
The Social Adjustment Scale Self-report (SAS-SR) was completed by chronic schizophrenics (N = 82) living in Nithsdale, in Dumfries and Galloway region. Relatives also assessed patients' adjustment. The questionnaire response indicated that scrutiny of the form by a professional immediately after completion is necessary. Patients' and relatives' assessment of the patients' overall adjustment correlated very closely, thus suggesting a schizophrenic can satisfactorily describe his own adjustment. The patients were less well adjusted than a normal community population in all areas except the parental role. The SAS-SR highlights for each individual where many of his problems lie, and therefore may act as a guide to specific rehabilitative measures.


Miller (Genee-Wiedemann) syndrome represents a clinically and biochemically distinct subgroup of postaxial acrofacial dysostosis associated with partial deficiency of DHODH.
Biallelic mutations in the gene encoding DHOdehase [dihydroorotate dehydrogenase (DHODH)], an enzyme required for de novo pyrimidine biosynthesis, have been identified as the cause of Miller (Genée-Weidemann or postaxial acrofacial dysostosis) syndrome (MIM 263750). We report compound heterozygous DHODH mutations in four additional families with typical Miller syndrome. Complementation in auxotrophic yeast demonstrated reduced pyrimidine synthesis and in vitro enzymatic analysis confirmed reduced DHOdehase activity in 11 disease-associated missense mutations, with 7 alleles showing discrepant activity between the assays. These discrepancies are partly explained by the domain structure of DHODH and suggest both assays are useful for interpretation of individual alleles. However, in all affected individuals, the genotype predicts that there should be significant residual DHOdehase activity. Urine samples obtained from two mutation-positive cases showed elevated levels of orotic acid (OA) but not dihydroorotate (DHO), an unexpected finding since these represent the product and the substrate of DHODH enzymatic activity, respectively. Screening of four unrelated cases with overlapping but atypical clinical features showed no mutations in either DHODH or the other de novo pyrimidine biosynthesis genes (CAD, UMPS), with these cases also showing normal levels of urinary OA and DHO. In situ analysis of mouse embryos showed Dhodh, Cad and Umps to be strongly expressed in the pharyngeal arch and limb bud, supporting a site- and stage-specific requirement for de novo pyrimidine synthesis. The developmental sensitivity to reduced pyrimidine synthesis capacity may reflect the requirement for an exceptional mitogenic response to growth factor signalling in the affected tissues.


Immediate autografting of bone in open fractures with bone loss of the hand: a preliminary report. Case reports.
Three patients with open fractures of the hand associated with bone loss were treated within four to six hours of injury by corticocancellous bone grafting and soft tissue coverage after meticulous debridement, copious irrigation of the wounds, and broad-spectrum antibiotics given intravenously. Long term follow-up was uneventful and showed that the graft had taken and healed well with early and full restoration of function and a good cosmetic result. Immediate corticocancellous bone grafting of an injured hand could be used in selected cases with well-debrided, surgically clean wounds as long as there is a rich blood supply. Adequate bone fixation, soft tissue coverage, and broad-spectrum antibiotics given intravenously will remove the risk of infection. Hand architecture is corrected while wound contracture and secondary deformity are avoided. Both patients' discomfort and hospital costs are considerably reduced.


Routine clean intermittent self-catheterization: innovative implementation in the Hunter Area Health Service.
Clean intermittent self-catheterization (CISC) has been used successfully for many years to promote socially acceptable and functional continence practice. The impact on health services, quality of life, and associated costs has also been documented. The use of CISC as a routine technique has been widely implemented in the Hunter Area Health Service, NSW, Australia. The positive outcomes resulting from the routine use of CISC has included reduction in length of stay and numbers of admissions for patients with urinary retention, reduction in nursing time, and decreased infection rates. The technique has been well received by patients who report a positive impact on their quality of life.


Sugar transport through maltoporin of Escherichia coli: role of the greasy slide.
The lining of the maltodextrin-specific maltoporin (LamB) channel exhibits a string of aromatic residues, the greasy slide, part of which has been shown previously by crystallography to be involved in substrate binding. To probe the functional role of the greasy slide, alanine scanning mutagenesis has been performed on the six greasy slide residues and Y118 at the channel constriction. The mutants were characterized by an in vivo uptake assay and sugar-induced-current-noise analysis. Crystallographic analysis of the W74A mutant showed no perturbation of the structure. All mutants showed considerably decreased maltose uptake rates in vivo (<10% of the wild-type value), indicating the functional importance of the investigated residues. Substitutions at the channel center revealed appreciably increased (up to 100-fold) in vitro half-saturation concentrations for maltotriose and maltohexaose binding to the channel. Sugar association rates, however, were significantly affected also by the mutations at either end of the slide (W74A, W358A, and F227A), an effect which became most apparent upon nonsymmetrical sugar addition. The kinetic data are discussed on the basis of an asymmetric one-site two-barrier model, which suggests that, at low substrate concentrations, as are found under physiological conditions, only the heights of the extracellular and periplasmic barriers, which are reduced by the presence of the greasy slide, determine the efficiency of this facilitated diffusion channel.


Diagnostic accuracy of 64-slice computed tomography coronary angiography for the detection of in-stent restenosis: a meta-analysis.
We sought to evaluate the diagnostic accuracy of 64-slice multi-detector row computed tomography (MDCT) compared with invasive coronary angiography for in-stent restenosis (ISR) detection. MEDLINE, Cochrane library, and BioMed Central database searches were performed until April 2009 for original articles. Inclusion criteria were (1) 64-MDCT was used as a diagnostic test for ISR, with >50% diameter stenosis selected as the cut-off criterion for significant ISR, using invasive coronary angiography and quantitative coronary angiography as the standard of reference; (2) absolute numbers of true positive, false positive, true negative, and false negative results could be derived. Standard meta-analytic methods were applied. Nine studies with a total of 598 patients with 978 stents included were considered eligible. On average, 9% of stents were unassessable (range 0-42%). Accuracy tests with 95% confidence intervals (CIs) comparing 64-MDCT vs invasive coronary angiography showed that pooled sensitivity, specificity, positive and negative likelihood ratio (random effect model) values were: 86% (95% CI 80-91%), 93% (95% CI 91-95%), 12.32 (95% CI 7.26-20.92), 0.18 (95% CI 0.12-0.28) for binary ISR detection. The symmetric area under the curve value was 0.94, indicating good agreement between 64-MDCT and invasive coronary angiography. 64-MDCT has a good diagnostic accuracy for ISR detection with a particularly high negative predictive value. However, still a relatively large proportion of stents remains uninterpretable. Accordingly, only in selected patients, 64-MDCT may serve as a potential alternative noninvasive method to rule out ISR.