Identification of parameters required for efficient lentiviral vector transduction and engraftment of human cord blood CD34(+) NOD/SCID-repopulating cells.
Human cord blood (CB) is a potential source of hematopoietic stem cells (HSC) for gene therapy to treat patients with hematopoietic disorders. However, limited numbers of CB CD34(+) cells, low transduction efficiency with lentiviral vectors (LVs), and low engraftment efficiency of nonobese diabetic/severe combined immunodeficient (NOD/SCID) repopulating cells (SRC), a measure of HSC, are blocks to this procedure. To optimize culture and transduction conditions, we compared various lengths of time for prestimulation before transduction, transduction duration, and posttransduction cell culture. We used a LV to transduce human CB CD34(+) cells followed by engraftment into NOD/SCID mice. We evaluated the effects of prestimulation and transduction time and optimized ex vivo cell culture duration before transplantation. We were able to achieve up to 40% transduction efficiency and up to 50% engraftment efficiency of SRC in CB CD34(+) cells when CB CD34(+) cells were either not prestimulated or prestimulated in 1% fetal bovine serum medium for 1 hour, followed by 5 hours transduction and 3 days culture in a cocktail of growth factors after transduction. No apparent functional changes of CB CD34(+) cells were noted under these conditions. This gene-transduction/cell-expansion protocol is the first systematic study to optimize prestimulation time, transduction time, and, very importantly, ex vivo culture time after transduction, and may be of use for LV gene transduction in a gene therapy setting.


Expression of M type 12 protein by a group A streptococcus exhibits phaselike variation: evidence for coregulation of colony opacity determinants and M protein.
Three major categories of colony opacity were observed for natural variants of the M type 12 (M12) group A streptococcus strain CS24. Colony opacity variants that switched between two alternative categories at significantly high frequencies were identified and are referred to as switching between more opaque (Op+) and less opaque (Op-) phenotypes. Twenty lineages of such variants were derived for analysis and were assessed for resistance to phagocytosis, acid-extractable M12 antigen, and M12 mRNA, criteria which define the M protein-positive phenotype (M+). Transition from the M+ to the M protein-negative phenotype (M-) correlated with a change from Op+ to Op-. Reversion to the Op+ phenotype was accompanied by reversion to the M+ state in all variants except one and occurred at a higher frequency than the forward M+ to M- switch. These data demonstrate the existence of M12 protein phaselike switching in the group A streptococcus strain CS24. The discovery of an Op+ M- revertant confirmed that colony opacity and M protein can be expressed independently and are distinct gene products. We suggest that coregulation of colony opacity and M protein expression accounts for their association among descendents of strain CS24. Southern blot hybridization analyses of digested genomic DNA from 27 M- variants and 15 M+ revertants were performed with DNA probes containing M12 protein and adjacent upstream sequences. DNA deletions were identified only in two stable M- variants, approximately 1.3 and 1.4 kilobases upstream from the M12 gene, respectively, whereas all unstable M- variants lacked detectable rearrangements. This suggests that deletions within or adjacent to the structural gene are unlikely to be responsible for the reversible switch in M protein expression. However, the association with the stable M- phenotype and the location of these deletions, as well as two other deletions, approximately 0.5 kilobase upstream from the M12 promoter in two previously described variants of strain CS24 suggests that a second gene product is required for full expression of M12 protein synthesis in this strain.


Zolpidem and promethazine in pre-anaesthetic medication. A pharmacopsychological approach.
This study dealt with the question: What are the effects of different doses of the benzodiazepine-like agent zolpidem and the phenothiazine derivative promethazine on mood in pre-anaesthetic medication? Subjects were 192 female and male patients awaiting elective surgery. Two drugs were administered to the patients in the evening before anaesthesia according to a 3 x 2 factorial randomized double-blind design. The first drug factor was zolpidem (8.03 vs. 16.06 mg) versus placebo. These drugs were combined with the second drug factor, either promethazine (50 mg) or placebo. In summary, the results confirm the hypothesis that zolpidem 8.03 mg can induce paradoxical effects, especially of anxiety, 1 h after application. When the dosage is doubled, no adverse effects are found, but there are no effects different from placebo either. Men could probably be described as a risk population for the adverse effects of zolpidem 8.03 mg. Furthermore, selective deactivating effects of promethazine 50 mg were shown in this study. In addition to these anaesthesiological aspects, this study showed promethazine to be useful as a research tool, suitable as a reference drug to detect deactivating effects. The multidimensional self-report inventory BSKE(EWL) (Befindlichkeitsskalierung durch Kategorien und Eigenschaftswörter) detected the expected drug effects, indicating that this instrument is suitable for further pharmacopsychological research.


Analysis of hospital ability to provide trauma services: a comparison between teaching and community hospitals.
Teaching hospitals (TH) can maintain the American College of Surgeons Committee on Trauma (ACSCOT) criteria for Level II trauma care more consistently than can community hospitals (CH). A retrospective analysis of 2,091 trauma system patients was done to determine if TH in an urban area are better able to meet the criteria for Level II trauma care than are CH. During the study period, a voluntary trauma plan existed among five hospitals; two TH and three CH. A hospital could accept patients that met trauma system entry criteria as long as, at that moment, it could provide the resources specified by ACSCOT. Hospitals were required to report their current resources accurately. A centralized communications center maintained a computerized, inter-hospital link which continuously monitored the availability of all participating hospitals. Trauma system protocols required paramedics to transport system patients to the closest available trauma hospital that had all the required resources available. Nine of the required ACSCOT Level II trauma center criteria were monitored for each institution emergency department (ED); trauma surgeon (TS); operating room (OR); angiography (ANG); anesthesiologist (ANE); intensive care unit (ICU); on-call surgeon (OCS); neurosurgeon (NS); and CT scanner (CT) available at the time of each trauma system entry. With the exception of OR, TH generally maintained the required staff and services more successfully than did CH. Further, less day to night variation in the available resources occurred at the TH. Specifically, ANE, ICU, TS, NS, and CT were available more often both day and night, at TH than CH. However, OR was less available at TH than CH during both day and night (p less than .01). In this community, TH provided a greater availability of trauma services than did CH. This study supports the designation of TH as trauma centers. A similar availability can be performed in other communities to help guide trauma center designation.


Does urinary incontinence affect middle-aged and older women's time use and activity patterns?
To investigate the relationship between urinary incontinence and women's levels and hours of participation in 31 activities. A subset of panel members from the Health and Retirement Study completed the self-administered Consumption and Activities Mail Survey questionnaire in 2001. These data were linked with Health and Retirement Study 2000 data. Analyses were limited to 2,190 female Consumption and Activities Mail Survey self-respondents born in 1947 or earlier. Logistic regression was used to predict activity participation. Linear regression was used to predict the number of hours of participation. The hypothesis that urinary incontinence affects women's time use and activity patterns was supported. Compared with the continent women, the incontinent women were less likely to have house cleaned, shopped, physically shown affection, or attended religious services in the recent past; and were more likely to have watched television or made music by singing or playing an instrument. Compared with continent activity participants, incontinent participants reported significantly fewer hours spent walking, communicating with friends and family by telephone or e-mail, working for pay, using a computer, and engaging in personal grooming and hygiene. These findings substantiate prior work on the relationship between urinary incontinence and quality of life, and suggest a useful route for educating patients about the impact of urinary incontinence. Clinicians must be alert to opportunities for encouraging incontinent women to be active. It is also important to consider the implications for time use and activity patterns when advising patients about treatment and management options. II-2.


Challenges in health and health care for Australia.
The next Australian Government will confront major challenges in the funding and delivery of health care. These challenges derive from: Changes in demography and disease patterns as the population ages, and the burden of chronic illness grows; Increasing costs of medical advances and the need to ensure that there are comprehensive, efficient and transparent processes for assessing health technologies; Problems with health workforce supply and distribution; Persistent concerns about the quality and safety of health services; Uncertainty about how best to balance public and private sectors in the provision and funding of health services; Recognition that we must invest more in the health of our children; The role of urban planning in creating healthy and sustainable communities; and Understanding that achieving equity in health, especially for Indigenous Australians, requires more than just providing health care services. The search for effective and lasting solutions will require a consultative approach to deciding the nation's priority health problems and to designing the health system that will best address them; issues of bureaucratic and fiscal responsibility can then follow.


Framewise multi-echo distortion correction for superior functional MRI.
Functional MRI (fMRI) data are severely distorted by magnetic field (B0) inhomogeneities which currently must be corrected using separately acquired field map data. However, changes in the head position of a scanning participant across fMRI frames can cause changes in the B0 field, preventing accurate correction of geometric distortions. Additionally, field maps can be corrupted by movement during their acquisition, preventing distortion correction altogether. In this study, we use phase information from multi-echo (ME) fMRI data to dynamically sample distortion due to fluctuating B0 field inhomogeneity across frames by acquiring multiple echoes during a single EPI readout. Our distortion correction approach, MEDIC (Multi-Echo DIstortion Correction), accurately estimates B0 related distortions for each frame of multi-echo fMRI data. Here, we demonstrate that MEDIC's framewise distortion correction produces improved alignment to anatomy and decreases the impact of head motion on resting-state functional connectivity (RSFC) maps, in higher motion data, when compared to the prior gold standard approach (i.e., TOPUP). Enhanced framewise distortion correction with MEDIC, without the requirement for field map collection, furthers the advantage of multi-echo over single-echo fMRI.


Simple oligomers as antimicrobial peptide mimics.
New approaches to antibiotic design are desperately needed. The design of simple oligomers that capture the shape and biological function of natural antimicrobial peptides could prove to be versatile and highly successful. We discuss the use of aromatic backbones to design facially amphiphilic (FA) beta-sheet like structures which are potently antimicrobial. These oligomers capture the physiochemical properties of peptides like the Magainins and Defensins, which fold into specific conformations that are amphiphilic resulting in antimicrobial activity. However, natural peptides are expensive to prepare and difficult to produce on large scale. The design of polymers and oligomers that mimic the complex structures and remarkable biological properties of proteins is an important endeavor and provides attractive alternatives to the difficult synthesis of natural peptides. We therefore have designed a series of FA oligomers that are easy to prepare from inexpensive monomers. They adopt structures very reminiscent of amphiphilic beta-sheets and have significant activity with minimal inhibitory concentrations at 6 h in the low microgram per ml range (muM to nM). They are active against a broad spectrum of bacteria including gram-positive and gram-negative as well as antibiotic resistant strains.


Cascade catalysis--strategies and challenges en route to preparative synthetic biology.
Nature's smartness and efficient assembling cascade type reactions inspired biologists and chemists all around the world. Tremendous effort has been directed towards the understanding and mimicking of such networks. In recent years considerable progress has been made in developing multistep one-pot reactions combining either advantage of chemo-, regio-, and stereoselectivity of biocatalysts or promiscuity and productivity of chemocatalysts. In this context several concepts, inspired by different disciplines (biocatalysis, metabolic engineering, synthetic chemistry, and material science), have been evolved. This review will focus on major contributions in the field of cascade reactions over the last three years.


The Importance of Ultrasound Monitoring of the Normal and Lesional Cervical Ectropion Treatment.
to investigate the importance of various ultrasound prognosis features in the assessment of the cervical ectropion treatment monitoring. The inclusion criteria was the presence of ectropion and the selection was based on clinical examination performed during routine consultations in specialized clinics, later confirmed by colposcopic evaluation of cervix. The evaluation protocol included: clinical evaluation completed with colposcopy, guided biopsy when lesions were suspected, serological assay of day 21 progesteronemy, presence of Chlamydia, Mycoplasma, Ureaplasma, HVS type II, HPV and bacterial infections, transvaginal ultrasound serial evaluation at the 7th, 14th and 21st day before and after tretment concerning: cervical volumetric calculations and velocimetric measurements of uterine arteries flows. Progestative treatment was prescribed, and antiinfectious specific treatment when needed. Patients were reevaluated after 3 months. The prospective study included 45 patients between 2013-2014. 28 presented serum progesterone levels below the reference range or borderline. We noted a moderate reduction of the ectropion area in 42 % and a marked reduction in 58% of the cases. No statistically significant differences were found between the size of the cervix before or after treatment, except certain evaluations (the 7th and the 14th day) in the presence of bacterial coinfections. Evaluation of pulsed Doppler velocimetric indices of uterine arteries flows showed generally minor variations with no constant positive or negative trend. Based on the data obtained in our study, we conclude that ultrasound monitoring of ectropion treatment do not provide reliable prognosis data regarding the evolution of cervical lesion.


Ambiguity aversion, modern Bayesianism and small worlds.
The central question of this paper is whether a rational agent under uncertainty can exhibit ambiguity aversion (AA). The answer to this question depends on the way the agent forms her probabilistic beliefs: classical Bayesianism (CB) vs modern Bayesianism (MB). We revisit Schmeidler's coin-based example and show that a rational MB agent operating in the context of a "small world", cannot exhibit AA. Hence we argue that the motivation of AA based on Schmeidler's coin-based and Ellsberg's classic urn-based examples, is poor, since they correspond to cases of "small worlds". We also argue that MB, not only avoids AA, but also proves to be normatively superior to CB because an MB agent (i) avoids logical inconsistencies akin to the relation between her subjective probability and objective chance, (ii) resolves the problem of "old evidence" and (iii) allows psychological detachment from actual evidence, hence avoiding the problem of "cognitive dissonance". As far as AA is concerned, we claim that it may be thought of as a (potential) property of large worlds, because in such worlds MB is likely to be infeasible.


Multiple scalp metastases from colonic neuroendocrine carcinoma: case report and literature review.
Colonic neuroendocrine neoplasms (NENs) are relatively rare tumors with an incidence rate of 0.11-0.21/100,000. NENs account for approximately 0.4% of colorectal neoplasms. Cutaneous metastases of colonic neuroendocrine carcinomas (NECs) are very infrequent, while cases of scalp metastasis are even fewer. Cutaneous metastases are more rare than visceral metastases and usually develop later; therefore, cutaneous metastases as initial distant metastases can be easily overlooked. This is the second case report of a colonic NEC with scalp metastasis. Compared with the previous case, in this instance scalp metastasis developed before visceral metastasis, and the cutaneous lesions were confined to the scalp alone. A 62-year-old Chinese man, who had undergone radical surgery for a "locoregional" colonic NEC one and half months before, came to our hospital for adjuvant chemotherapy. We found multiple scalp nodules during physical examination. Moreover, these nodules had occurred and had not been detected prior to the patient undergoing radical surgery. The scalp nodules proved to be metastases from colonic NEC as determined using pathological and immunohistochemical examinations following lumpectomy. After one and half months, visceral metastases were detected in this patient. Ultimately, the patient died two months later. In this report an unusual case of a colonic NEC with initial distant metastasis confined to the scalp is presented. This case is unusual because of the development of cutaneous metastasis before visceral metastasis. The scalp metastasis were initially overlooked, leading to inaccurate staging and radical surgery that was not curative. This demonstrates that distant metastasis can occur during the early phase of tumor growth in these aggressive lesions. Thus, the possibility of distant metastases should be assessed in the initial work up to avoid mistaken clinical staging especially when distant metastases occur only in skin.


Statistical parametric maps of ¹⁸F-FDG PET and 3-D autoradiography in the rat brain: a cross-validation study.
Although specific positron emission tomography (PET) scanners have been developed for small animals, spatial resolution remains one of the most critical technical limitations, particularly in the evaluation of the rodent brain. The purpose of the present study was to examine the reliability of voxel-based statistical analysis (Statistical Parametric Mapping, SPM) applied to (18)F-fluorodeoxyglucose (FDG) PET images of the rat brain, acquired on a small animal PET not specifically designed for rodents. The gold standard for the validation of the PET results was the autoradiography of the same animals acquired under the same physiological conditions, reconstructed as a 3-D volume and analysed using SPM. Eleven rats were studied under two different conditions: conscious or under inhalatory anaesthesia during (18)F-FDG uptake. All animals were studied in vivo under both conditions in a dedicated small animal Philips MOSAIC PET scanner and magnetic resonance images were obtained for subsequent spatial processing. Then, rats were randomly assigned to a conscious or anaesthetized group for postmortem autoradiography, and slices from each animal were aligned and stacked to create a 3-D autoradiographic volume. Finally, differences in (18)F-FDG uptake between conscious and anaesthetized states were assessed from PET and autoradiography data by SPM analysis and results were compared. SPM results of PET and 3-D autoradiography are in good agreement and led to the detection of consistent cortical differences between the conscious and anaesthetized groups, particularly in the bilateral somatosensory cortices. However, SPM analysis of 3-D autoradiography also highlighted differences in the thalamus that were not detected with PET. This study demonstrates that any difference detected with SPM analysis of MOSAIC PET images of rat brain is detected also by the gold standard autoradiographic technique, confirming that this methodology provides reliable results, although partial volume effects might make it difficult to detect slight differences in small regions.


Accessory subunits are integral for assembly and function of human mitochondrial complex I.
Complex I (NADH:ubiquinone oxidoreductase) is the first enzyme of the mitochondrial respiratory chain and is composed of 45 subunits in humans, making it one of the largest known multi-subunit membrane protein complexes. Complex I exists in supercomplex forms with respiratory chain complexes III and IV, which are together required for the generation of a transmembrane proton gradient used for the synthesis of ATP. Complex I is also a major source of damaging reactive oxygen species and its dysfunction is associated with mitochondrial disease, Parkinson's disease and ageing. Bacterial and human complex I share 14 core subunits that are essential for enzymatic function; however, the role and necessity of the remaining 31 human accessory subunits is unclear. The incorporation of accessory subunits into the complex increases the cellular energetic cost and has necessitated the involvement of numerous assembly factors for complex I biogenesis. Here we use gene editing to generate human knockout cell lines for each accessory subunit. We show that 25 subunits are strictly required for assembly of a functional complex and 1 subunit is essential for cell viability. Quantitative proteomic analysis of cell lines revealed that loss of each subunit affects the stability of other subunits residing in the same structural module. Analysis of proteomic changes after the loss of specific modules revealed that ATP5SL and DMAC1 are required for assembly of the distal portion of the complex I membrane arm. Our results demonstrate the broad importance of accessory subunits in the structure and function of human complex I. Coupling gene-editing technology with proteomics represents a powerful tool for dissecting large multi-subunit complexes and enables the study of complex dysfunction at a cellular level.


Free radical-mediated oxidation of free amino acids and amino acid residues in proteins.
We summarize here results of studies designed to elucidate basic mechanisms of reactive oxygen (ROS)-mediated oxidation of proteins and free amino acids. These studies have shown that oxidation of proteins can lead to hydroxylation of aromatic groups and aliphatic amino acid side chains, nitration of aromatic amino acid residues, nitrosylation of sulfhydryl groups, sulfoxidation of methionine residues, chlorination of aromatic groups and primary amino groups, and to conversion of some amino acid residues to carbonyl derivatives. Oxidation can lead also to cleavage of the polypeptide chain and to formation of cross-linked protein aggregates. Furthermore, functional groups of proteins can react with oxidation products of polyunsaturated fatty acids and with carbohydrate derivatives (glycation/glycoxidation) to produce inactive derivatives. Highly specific methods have been developed for the detection and assay of the various kinds of protein modifications. Because the generation of carbonyl derivatives occurs by many different mechanisms, the level of carbonyl groups in proteins is widely used as a marker of oxidative protein damage. The level of oxidized proteins increases with aging and in a number of age-related diseases. However, the accumulation of oxidized protein is a complex function of the rates of ROS formation, antioxidant levels, and the ability to proteolytically eliminate oxidized forms of proteins. Thus, the accumulation of oxidized proteins is also dependent upon genetic factors and individual life styles. It is noteworthy that surface-exposed methionine and cysteine residues of proteins are particularly sensitive to oxidation by almost all forms of ROS; however, unlike other kinds of oxidation the oxidation of these sulfur-containing amino acid residues is reversible. It is thus evident that the cyclic oxidation and reduction of the sulfur-containing amino acids may serve as an important antioxidant mechanism, and also that these reversible oxidations may provide an important mechanism for the regulation of some enzyme functions.


Complications in Brow Lifts: A Systemic Review of Surgical and Nonsurgical Brow Rejuvenations.
Brow lift has been a part of plastic surgeons' armamentarium for over a century. Although there are multiple different techniques available, there has been a dearth of literature comparing which technique is better and has lesser complications. In this study, we performed a systemic literature review of all brow lift techniques to determine and compare complication rates of each technique. We performed a computerized search of brow rejuvenation techniques using the MEDLINE database. Data regarding the type of brow lift procedure, number of patients, and complications were collected. The systematic review was performed in December of 2017. A total of 326 articles were identified initially, and 76 studies remained after final review. The review showed highest revision rate in the hairline brow lift (7.4%), highest numbness rate in the direct brow lift (5.5%), highest asymmetry rate in the temporal/lateral brow lift (1.5%), and highest alopecia rate in the endoscopic brow lift (2.8%). In addition, we found 5 studies in nonsurgical brow rejuvenation (3 botulinum toxin, 1 fat injection, 1 radiofrequency). Our study shows that each brow technique has a different complication profile, and it is important for plastic surgeons to understand the difference. Furthermore, the study demonstrated the lack of randomized prospective outcome studies and standardization of outcome measures in brow rejuvenation. We believe this information would be the key and next frontier in providing technically safe and long-lasting aesthetic outcomes for the patients.


In vitro bone marrow purging of multidrug-resistant cells with a mouse monoclonal antibody directed against Mr 170,000 glycoprotein and a saporin-conjugated anti-mouse antibody.
Selective elimination of multidrug resistance-positive cells (LoVo/Dx) was obtained by using the monoclonal antibody MRK 16, which recognizes a surface epitope of the Mr 170,000 glycoprotein, and a sheep anti-mouse immunoglobulin antibody, conjugated to the ribosome-inactivating protein saporin 6. The killing was greatly decreased or even abolished by adding the monoclonal antibody at a 100-fold concentration. Both the MRK 16 and anti-mouse saporin 6 conjugate did not show any killing activity when they were used separately. In cell suspensions composed of 90% normal bone marrow cells and 10% multidrug resistance-positive cells, the monoclonal antibody MRK 16 followed by the anti-mouse immunotoxin caused the elimination of 99% multidrug resistance-positive cells, as revealed by immunofluorescence and immunocytochemistry as well as by a clonal assay. Human normal hematopoietic precursors (granulomonocytic colony-forming units, erythroid burst-forming units, and multipotent granulomonocytic, erythroid, and megakaryocytic-forming units) were not affected by the MRK 16 plus immunotoxin treatment. This technique might be suitable for ex vivo bone purging in an appropriate clinical setting, such as autologous bone marrow transplantation.


Soybean seed acid phosphatases: unusual optimum temperature and thermal stability studies.
In contrast to other acid phosphatases, four cytoplasmic isoforms (AP1, AP2, AP3A, and AP3B) purified from mature soybean seeds presented high activities at temperatures above 80 degrees C, when p-nitrophenylphosphate (p-NPP) was utilized as substrate. However, with tyrosine phosphate and inorganic pyrophosphate as substrates, maximum activities were observed at temperature of 60 degrees C during 10 min reaction. In the absence of substrate, enzymes lost only 20% activity after 60 min at 60 degrees C; the isoforms AP3A and AP3B retained 30% of activity at 70 degrees C after 60 min and all the isoforms were inactivated at 80 degrees C, after 5 min. Thermal inactivation studies indicated that the soybean enzymes showed different temperature dependences in relation to most plant acid phosphatases. A best protective effect was observed when the isoforms were preincubated, at 70 degrees C, with phosphate (10 mM) and p-nitrophenol (10 mM) which indicates that the enzyme inactivation was prevented only in the presence of both reaction products.


Generation of influenza A virus NS2 (NEP) mutants with an altered nuclear export signal sequence.
The NS2 (NEP) protein of influenza A virus contains a highly conserved nuclear export signal (NES) motif in its amino-terminal region (12ILMRMSKMQL21, A/WSN/33), which is thought to be required for nuclear export of viral ribonucleoprotein complexes (vRNPs) mediated by a cellular export factor, CRM1. However, simultaneous replacement of three hydrophobic residues in the NES with alanine does not affect NS2 (NEP) binding to CRM1, although the virus with these mutations is not viable. To determine the extent of sequence conservation required by the NS2 (NEP) NES for its export function during viral replication, we randomly introduced mutations by degenerative mutagenesis into the region of NS cDNA encoding the NS2 (NEP) NES and then attempted to generate mutant viruses containing these alterations by reverse genetics. Sequence analysis of the recovered viruses showed that although some of the mutants possessed amino acids other than those conserved in the NES, hydrophobicity within this motif was maintained. Nuclear export of vRNPs representing all of the mutant viruses was completely inhibited in the presence of a CRM1 inhibitor, leptomycin B, as was the transport of wild-type virus, indicating that the CRM1-mediated pathway is responsible for the nuclear export of both wild-type and mutant vRNPs. The vRNPs of some of the mutant viruses were exported in a delayed manner, resulting in limited viral growth in cell culture and in mice. These results suggest that the NES motif may be an attractive target for the introduction of attenuating mutations in the production of live vaccine viruses.


Association mapping by generalized linear regression with density-based haplotype clustering.
Haplotypes of closely linked single-nucleotide polymorphisms (SNPs) potentially offer greater power than individual SNPs to detect association between genetic variants and disease. We present a novel approach for association mapping in which density-based clustering of haplotypes reduces the dimensionality of the general linear model (GLM)-based score test of association implemented in the HaploStats software (Schaid et al. [2002] Am. J. Hum. Genet. 70:425-434). A flexible haplotype similarity score, a generalization of previously used measures, forms the basis, for grouping haplotypes of probable recent common ancestry. All haplotypes within a cluster are assigned the same regression coefficient within the GLM, and evidence for association is assessed with a score statistic. The approach is applicable to both binary and continuous trait data, and does not require prior phase information. Results of simulation studies demonstrated that clustering enhanced the power of the score test to detect association, under a variety of conditions, while preserving valid Type-I error. Improvement in performance was most dramatic in the presence of extreme haplotype diversity, while a slight improvement was observed even at low diversity. Our method also offers, for binary traits, a slight advantage in power over a similar approach based on an evolutionary model (Tzeng et al. [2006] Am. J. Hum. Genet. 78:231-242).


Childhood sexual abuse and sexual risk behavior among men and women attending a sexually transmitted disease clinic.
Childhood sexual abuse (CSA) is associated with a wide range of negative outcomes. The authors investigated the relation between CSA and sexual risk behavior in 827 patients recruited from a sexually transmitted disease (STD) clinic. Overall, CSA was reported by 53% of women and 49% of men and was associated with greater sexual risk behavior, including more sexual partners, unprotected sex, and sex trading. Alcohol use for men and drug use for women mediated the relation between CSA and the number of sexual partners in the past 3 months; intimate partner violence mediated the relation between CSA and the number of episodes of unprotected sex in the past 3 months for women. These results document the prevalence of CSA among patients seeking care for an STD and can be used to tailor sexual risk reduction programs for individuals who were sexually abused.


Mathematical modeling at the livestock-wildlife interface: scoping review of drivers of disease transmission between species.
Modeling of infectious diseases at the livestock-wildlife interface is a unique subset of mathematical modeling with many innate challenges. To ascertain the characteristics of the models used in these scenarios, a scoping review of the scientific literature was conducted. Fifty-six studies qualified for inclusion. Only 14 diseases at this interface have benefited from the utility of mathematical modeling, despite a far greater number of shared diseases. The most represented species combinations were cattle and badgers (for bovine tuberculosis, 14), and pigs and wild boar [for African (8) and classical (3) swine fever, and foot-and-mouth and disease (1)]. Assessing control strategies was the overwhelming primary research objective (27), with most studies examining control strategies applied to wildlife hosts and the effect on domestic hosts (10) or both wild and domestic hosts (5). In spatially-explicit models, while livestock species can often be represented through explicit and identifiable location data (such as farm, herd, or pasture locations), wildlife locations are often inferred using habitat suitability as a proxy. Though there are innate assumptions that may not be fully accurate when using habitat suitability to represent wildlife presence, especially for wildlife the parsimony principle plays a large role in modeling diseases at this interface, where parameters are difficult to document or require a high level of data for inference. Explaining observed transmission dynamics was another common model objective, though the relative contribution of involved species to epizootic propagation was only ascertained in a few models. More direct evidence of disease spill-over, as can be obtained through genomic approaches based on pathogen sequences, could be a useful complement to further inform such modeling. As computational and programmatic capabilities advance, the resolution of the models and data used in these models will likely be able to increase as well, with a potential goal being the linking of modern complex ecological models with the depth of dynamics responsible for pathogen transmission. Controlling diseases at this interface is a critical step toward improving both livestock and wildlife health, and mechanistic models are becoming increasingly used to explore the strategies needed to confront these diseases.


Mobility training using a bionic knee orthosis in patients in a post-stroke chronic state: a case series.
An emerging area of neurorehabilitation is the use of robotic devices to enhance the efficiency and effectiveness of lower extremity physical therapy post-stroke. Many of the robotic devices currently available rely on computer-driven locomotive algorithms combined with partial bodyweight-supported treadmill training that drive reflex stepping with minimal patient intention during therapy. In this case series, we examined the effect of task-oriented mobility training in patients in a post-stroke chronic state using a novel, wearable, mobile, intention-based robotic leg orthosis. Three individuals, all of whom had reached a plateau with conventional bodyweight-supported treadmill training, participated in task-oriented mobility therapy (1.5 hours, two to four times per week for four weeks) with a robotic leg orthosis under supervision by a physical therapist. Participant 1 was a 59-year-old Caucasian man, who had an ischemic left stroke six years previously with resultant right hemiparesis. Participant 2 was a 42-year-old Caucasian woman with left hemiparesis after a right stroke 15 months previously. Participant 3 was a 62-year-old Caucasian woman with a history of a right middle cerebral artery aneurysm with third degree sub-arachnoid hemorrhage 10 years ago.Immediately after training, all participants demonstrated improved gait speed (10 meter walk), stride length and walking endurance (6 minute walk) compared with baseline measurements. Improvements were maintained one month after training. Timed up and go and five times sit-to-stand were maintained for all three participants, with only one individual remaining outside the safety performance norm. Lower extremity training integrating an intention-based robotic leg orthosis may improve gait speed, endurance and community levels of participation in select patients in a post-stroke chronic state after plateauing within a bodyweight-supported treadmill training program. The wearable, mobile assistive robotic device safely supplemented supervised physical therapy including mobility and balance skill training.


[The patient's privacy and the treatment of intimacy: an ethical reflection].
Although no one today doubts that speaking of a nurse's patient care presupposes the nurse will carry out her/his role as a professional; nonetheless, this has been related to the patient's intimacy very few times. This article concerns this last perspective since the author maintains as her hypothesis that the sharing of myths and images people hold, as an essential element of each person, requires nurse's care. For this reason, the author presents the concept of the intimacy of the patient from an ethical point of view and defines the principles which compose the treatment of intimacy.


Early introduction of fish decreases the risk of eczema in infants.
Atopic eczema in infants has increased in western societies. Environmental factors and the introduction of food may affect the risk of eczema. To investigate the prevalence of eczema among infants in western Sweden, describe patterns of food introduction and assess risk factors for eczema at 1 year of age. Data were obtained from a prospective, longitudinal cohort study of infants born in western Sweden in 2003; 8176 families were randomly selected and, 6 months after the infant's birth, were invited to participate and received questionnaires. A second questionnaire was sent out when the infants were 12 months old. Both questionnaires were completed and medical birth register data were obtained for 4921 infants (60.2% of the selected population). At 1 year of age, 20.9% of the infants had previous or current eczema. Median age at onset was 4 months. In multivariable analysis, familial occurrence of eczema, especially in siblings (OR 1.87; 95% confidence interval (CI) 1.50 to 2.33) or the mother (OR 1.54; 95% CI 1.30 to 1.84), remained an independent risk factor. Introducing fish before 9 months of age (OR 0.76; 95% CI 0.62 to 0.94) and having a bird in the home (OR 0.35; 95% CI 0.17 to 0.75) were beneficial. One in five infants suffer from eczema during the first year of life. Familial eczema increased the risk, while early fish introduction and bird keeping decreased it. Breast feeding and time of milk and egg introduction did not affect the risk.


Preclinical detection of variant CJD and BSE prions in blood.
The emergence of variant Creutzfeldt Jakob Disease (vCJD) is considered a likely consequence of human dietary exposure to Bovine Spongiform Encephalopathy (BSE) agent. More recently, secondary vCJD cases were identified in patients transfused with blood products prepared from apparently healthy donors who later went on to develop the disease. As there is no validated assay for detection of vCJD/BSE infected individuals the prevalence of the disease in the population remains uncertain. In that context, the risk of vCJD blood borne transmission is considered as a serious concern by health authorities. In this study, appropriate conditions and substrates for highly efficient and specific in vitro amplification of vCJD/BSE agent using Protein Misfolding Cyclic Amplification (PMCA) were first identified. This showed that whatever the origin (species) of the vCJD/BSE agent, the ovine Q171 PrP substrates provided the best amplification performances. These results indicate that the homology of PrP amino-acid sequence between the seed and the substrate is not the crucial determinant of the vCJD agent propagation in vitro. The ability of this method to detect endogenous vCJD/BSE agent in the blood was then defined. In both sheep and primate models of the disease, the assay enabled the identification of infected individuals in the early preclinical stage of the incubation period. Finally, sample panels that included buffy coat from vCJD affected patients and healthy controls were tested blind. The assay identified three out of the four tested vCJD affected patients and no false positive was observed in 141 healthy controls. The negative results observed in one of the tested vCJD cases concurs with results reported by others using a different vCJD agent blood detection assay and raises the question of the potential absence of prionemia in certain patients.


Physical model of differential Mueller matrix for depolarizing uniform media.
In this article, we address the question of significance of the parameters of differential Mueller matrix formalism. We show how the concept of mean value and uncertainty of the optical properties recently introduced to depict this differential matrix can be related to the random fluctuations of these optical properties. From the layered-medium interpretation introduced by Jones [J. Opt. Soc. Am. 38, 671 (1948)] and extended to Mueller-Jones matrix by Azzam [J. Opt. Soc. Am. 68, 1756 (1978)], a generalization to depolarizing Mueller matrices is proposed. Based on the random Mueller-Jones matrix approach, the obtained parameterization perfectly fits the previous results from the literature. Necessary conditions of positivity on specific coefficients imposed in order to have physical Mueller matrix are introduced in a natural way and not inferred a posteriori. Interpretations of the underlying physical processes are also presented. An illustrative experimental example is provided from literature data.


Enhancement of Opioid Antinociception by Nicotine.
Nicotine can produce antinociception in preclinical pain models; however, the ability of nicotine to augment the antinociceptive effects of opioid agonists has not been investigated. The present experiments were conducted to determine how nicotine modifies the effects of opioid agonists differing in efficacy. Male squirrel monkeys responded for the delivery of milk under a fixed ratio 10 schedule of reinforcement. During the 30-second timeout period following each milk delivery, the subject's tail was immersed in 35, 50, 52, or 55°C water, and the latency to remove the tail was recorded. Dose-response functions for tail-withdrawal latency and operant performance were determined for fentanyl, oxycodone, buprenorphine, and nalbuphine alone and after treatment with nicotine. Excepting nalbuphine, all opioids produced dose-related disruptions in food-maintained responding and increases in tail-withdrawal latency at each water temperature. Nicotine did not exacerbate the behaviorally disruptive effects of the μ-opioids on operant performance but produced a significant mecamylamine-sensitive enhancement of the antinociceptive potency of each opioid. Failure of arecoline to augment the antinociceptive effects of oxycodone and antagonism by mecamylamine suggests this nicotine-induced augmentation of prescription opioid antinociception was nicotinic acetylcholine receptor (nAChR) mediated. This was reflected in leftward shifts in the antinociceptive dose-response curve of each opioid, ranging from 2- to 7-fold increases in the potency of oxycodone across all water temperatures to an approximately 70-fold leftward shift in the antinociceptive dose-response curve of nalbuphine at the lower and intermediate water temperatures. These results suggest that nicotine may enhance μ-opioid antinociceptive effects without concomitantly exacerbating their behaviorally disruptive effects. SIGNIFICANCE STATEMENT: Prescription opioids remain the most effective pain-management pharmacotherapeutics but are limited by their adverse effects. The present results indicate that nicotine enhances antinociceptive effects of various opioid agonists in nonhuman primates without increasing their disruptive effects on operant performance. These results suggest that nicotine might function as an opioid adjuvant for pain management by enabling decreased clinically effective analgesic doses of prescription opioids without exacerbating their adverse behavioral effects.


Cell and matrix reactions at titanium implants in surgically prepared rat tibiae.
The tissue response of rat tibiae to the surgical placement of commercially pure titanium implants was examined at 2, 6, 10, and 28 days. The transcortical placement of 1.5-mm x 2-mm implants resulted in the apposition of threaded implant surfaces within cortical and cancellous regions of the tibia. In all regions, evidence of bone formation was obtained through pre-embedding fracture of the implant from the bone tissue interface. Scanning electron microscopy examination of early responses revealed a fibrin clot and rapid formation of a loosely organized collagenous matrix. Many extravasated blood cells contacted the implant surface. At day 6, a more organized matrix containing many blood vessels opposed the implant surfaces, and few extravasated blood cells remained in contact with the implant surface. By day 10, the surgical wound was filled with woven bone that approximated the contours of the threaded implant. Later, few cells were attached to the retrieved implants. The consolidation of the forming matrix was clearly evident at 28 days. The tissue interface was an amorphous matrix that revealed the surface characteristics of the machined implant. Light microscopic analysis of ground sections indicated that, from day 6 onward, cells morphologically consistent with the osteoblastic phenotype were predominant within the gap between the surgical margin and implant surface. Osteoblastic cells had achieved the formation of an osteoid seam upon which bone formation progressed. The matrix that had formed represented woven bone containing many osteocytes. At day 6, evidence of remodeling was observed at sites distant from the surgical site, and by day 28 osteoclastic activity was observed at trabecular sites adjacent to the implant surface. The rat tibia model provides evidence of rapid formation of bone at implant surfaces.


Biological comparison between three clones of Trypanosoma cruzi and the strain of origin (Bolivia) with reference to clonal evolution studies.
After isolating three clones of Trypanosoma cruzi (Bolivia), we first characterized them according to parasitaemia, pleomorphism and virulence, and then histopathologically. The study's interest lies on the hypothesis that clonal evolution of T. cruzi has a major impact on biologically relevant properties of this parasite. Data obtained from the studies of parasitaemia, pleomorphism and virulence showed no differences between the groups studied. As a final point, the histopathological study shows us a muscular tissue tropism both in clones and in their mother strain (Bolivia). In this paper, we conclude that Bolivia strain and clones isolated from it, pertaining to the same major clone share similar biological properties.


[Immunogenic properties of various forms of flagellin].
The authors obtained and studied immunologically flagellae of S. typhi and flagellins (monomeric and polymeric) obtained from them. Immunological activity of flagellin was the greater the higher the molecular weight of the preparation (the polymer molecules were the largest and the most immunogenic). There were revealed differences between the preparations according to the dependence of the immunizing effect on the dose and the capacity to induce synthesis of 19S- and 7S-antibodies. A sharp change of the immunogenicity of flagellin preparation following gel-filtration through sepharose 2B was demonstrated.


Impaired urinary concentration after vasopressin and its gradual correction in hypothalamic diabetes insipidus.
This study utilized rates with hereditary hypothalamic diabetes insipidus (D.I.) in order to explore possible mechanisms which prevent full urinary concentration after acute administration of vasopressin in hypothalamic D.I. and which correct this concentrating defect with prolonged therapy.IT WAS FOUND: (a) that the concentrating defect persisted even when the urinary osmolal excretion of D. I. rats was reduced to that of normal animals; (b) that the defect was not corrected more rapidly if larger doses of vasopressin were given; (c) that it persisted even when the D.I. rats were deprived of drinking water after vasopressin was given; (d) that there was osmotic equilibration between urine and renal papilla at a time when the concentrating defect was still evident; and (e) that the correction of the defect was associated with progressive and significant rise of the papillary osmolality. These studies appear to rule out osmotic diuresis, accumulation of exogenous vasopressin, persistent primary polydipsia, or delay in the induction of membrane permeability as causes for the concentrating defect. Rather, subnormal osmolality of the renal papilla, which can be corrected only gradually, accounts for the initial concentrating defect and the long time required for its correction. Reduction of water content and increase of urea content are primarily responsible for restoration of papillary osmolality to normal.


Use of advanced technologies across the wound care spectrum: prologue.
The growing trends of ageing populations and increasing prevalence of diabetes have given rise to an expanding number of problematic acute and chronic wounds. Over the past two decades, the use of negative pressure wound therapy (NPWT) with and without instillation of topical wound solutions has expanded to include treatment of a large variety of wounds. Additionally, the use of NPWT specifically designed for use over closed surgical incisions has been associated with favourable results. The recent introduction of an automated epidermal harvesting system, which neither creates a donor site wound nor requires the use of a surgeon, operating room or anaesthesia, has facilitated the use of epidermal grafting. This supplement highlights some of the advanced wound approaches that have been developed to address challenging wounds and the growing burden of wound care affecting both the patient and the health care system.


[The efficacy of Piracetam on the mental functional capacity of chronic alcoholics (author's transl)].
Piracetam, 1-pyrrolidone acetamid, was tested in 40 chronic alcoholics with a more or less marked psycho-organic syndrome by means of psychological tests. It was a double-blind-cross-over study. Statistical analysis of the results showed that Piracetam improves the energo-functional capacity of the cortex i.e. the basal functions of the cortical cells such as activating capacity, vital dynamic, flexibility, intellectual reactivity and stress tolerance. Apart from the overall improvement we also observed an improvement of specific cerebral performances which is however unimportant in comparison with the greneralized effect.


Genome-Scale Metabolic Modelling of Lifestyle Changes in Rhizobium leguminosarum.
Biological nitrogen fixation in rhizobium-legume symbioses is of major importance for sustainable agricultural practices. To establish a mutualistic relationship with their plant host, rhizobia transition from free-living bacteria in soil to growth down infection threads inside plant roots and finally differentiate into nitrogen-fixing bacteroids. We reconstructed a genome-scale metabolic model for Rhizobium leguminosarum and integrated the model with transcriptome, proteome, metabolome, and gene essentiality data to investigate nutrient uptake and metabolic fluxes characteristic of these different lifestyles. Synthesis of leucine, polyphosphate, and AICAR is predicted to be important in the rhizosphere, while myo-inositol catabolism is active in undifferentiated nodule bacteria in agreement with experimental evidence. The model indicates that bacteroids utilize xylose and glycolate in addition to dicarboxylates, which could explain previously described gene expression patterns. Histidine is predicted to be actively synthesized in bacteroids, consistent with transcriptome and proteome data for several rhizobial species. These results provide the basis for targeted experimental investigation of metabolic processes specific to the different stages of the rhizobium-legume symbioses. IMPORTANCE Rhizobia are soil bacteria that induce nodule formation on plant roots and differentiate into nitrogen-fixing bacteroids. A detailed understanding of this complex symbiosis is essential for advancing ongoing efforts to engineer novel symbioses with cereal crops for sustainable agriculture. Here, we reconstruct and validate a genome-scale metabolic model for Rhizobium leguminosarum bv. viciae 3841. By integrating the model with various experimental data sets specific to different stages of symbiosis formation, we elucidate the metabolic characteristics of rhizosphere bacteria, undifferentiated bacteria inside root nodules, and nitrogen-fixing bacteroids. Our model predicts metabolic flux patterns for these three distinct lifestyles, thus providing a framework for the interpretation of genome-scale experimental data sets and identifying targets for future experimental studies.


Disentangling direct and indirect effects of experimental grassland management and plant functional-group manipulation on plant and leafhopper diversity.
Plant biodiversity can affect trophic interactions in many ways, including direct bottom-up effects on insects, but is negatively affected by agricultural intensification. Grassland intensification promotes plant productivity, resulting in changes in plant community composition, and impacts on higher trophic levels. Here, we use a novel grassland management experiment combining manipulations of cutting and fertilization with experimental changes in plant functional group composition (independent of management effects) to disentangle the direct and indirect effects of agricultural management on insect herbivore diversity and abundance. We used leafhoppers as model organisms as they are a key insect taxon in grasslands and react rapidly to management changes. Leafhoppers were sampled between May and September 2010 using standardized sweep netting and pan traps. Plant diversity, functional group composition and management regime in grasslands affected leafhopper species richness and abundance. Higher cutting frequencies directly led to decreasing leafhopper species richness, presumably due to the higher disturbance frequency and the reduction in food-resource heterogeneity. In contrast, fertilizer application had only a small indirect negative effect via enhanced aboveground plant biomass, reduced plant diversity and changes in functional group composition. The manipulated increase in grass cover had contrasting direct and indirect effects on leafhopper species richness: grass cover directly increased leafhopper species richness, but negatively affected plant diversity, which in turn was positively related to leafhopper species richness. In conclusion, insect diversity is driven in complex direct and indirect ways by grassland management, including changes in functional group composition. The availability of preferred food sources and the frequency of disturbance are important direct and indirect drivers of leafhopper species richness, interacting in complex ways with plant diversity and food resource heterogeneity.


Suppressive substance produced by T cells from mice chronically infected with Trypanosoma cruzi. I. Preferential inhibition of the induction of delayed-type hypersensitivity.
Culture supernatants of spleen cells from susceptible CBA mice chronically infected with Trypanosoma cruzi were able to inhibit the induction of delayed-type hypersensitivity (DTH) to a wide range of antigens as measured by 24-hr footpad swelling, bone marrow homing, and radioactivity accumulation assays. The suppressive activity, which was also present in the serum of these chronically infected mice, appears to be specific for the induction of DTH and had no effect on the 3-hr immediate-type hypersensitivity. It also failed to modify the expression of DTH in presensitized mice. Furthermore, it did not affect the synthesis in normal recipients of specific antibody or the induction of helper T cells or cytotoxic T cells. It also failed to induce DTH tolerance as recipient mice with markedly reduced DTH were able to develop a normal DTH response after secondary immunization. The suppressive activity was produced by an Ig- macrophage-depleted splenic T cell population, whose capacity to secrete the suppressive substance was completely abrogated by treatment in vitro with anti-L3T4 antibody and complement, but not with anti-Lyt-2 antibody and complement. These results therefore demonstrate that L3T4+ T cells from mice chronically infected with T. cruzi can produce substances which interfere with the induction of DTH. This finding may help to identify the differential antigenic stimulatory requirement for the activation of the various subsets of T cells.


Smoking prevalence, knowledge, and attitudes among a population of Vietnamese American men.
Tobacco use among Vietnamese adult males in the United States is higher than the general population. Less is known about the role of knowledge and attitudes of smoking in smoking status. This study describes the smoking prevalence, practices, support, knowledge, and attitudes among Vietnamese American men by smoking status. We administrated a cross-sectional in-person health questionnaire to randomly selected Vietnamese men (18-64 years of age) living in Seattle, Washington, using bilingual, bicultural Vietnamese male interviewers (N = 509). The response rate was 79%; the cooperation rate was 82%. Sixty-four percent of respondents had a history of smoking: 37% current, 27% former, and 36% never smokers. Smoking prevalence was lowest among men aged 18-29 years. Among smokers, 81% smoked 1 to 10 cigarettes per day, 69% wanted to quit, and 48% planned to do so in the next 6 months. Twelve percent of smokers reported smoking was allowed in the home. On average, respondents correctly answered six out of seven questions regarding health risks related to smoking. In logistic regression analyses, being a current smoker was negatively associated with a higher knowledge score (OR = 0.83, 95% CI 0.71-0.97). Adjusted odds of being a current smoker were 3.77 times higher among men who agreed with the attitude statement "It is appropriate for Vietnamese men to smoke when with friends." (OR = 2.15, 95% CI 1.28-3.61). The findings suggest a great need to develop appropriate tobacco-control interventions to lower smoking prevalence, improve tobacco-related health knowledge, and reduce the acceptance of smoking among Vietnamese American men.


Fine structure of synapses of the central nervous system in resinless sections.
The cytoskeleton has been implicated in neuronal function, particularly in axonal transport, excitability at axonal membranes, and movement of synaptic vesicles at preganglionic endings. The present study demonstrates the presence of a pre- and postsynaptic cytoskeleton in resinless sections of CNS tissue by use of the polyethylene glycol (PEG) technique of Wolosewick (1980) viewed by conventional transmission EM, scanning transmission EM, and surface scanning EM. The PEG technique permits visualization of the cytoskeletal network unobscured by the electron scattering properties of epoxy embedment. In the presynaptic process, synaptic vesicles appear to be suspended in a filamentous network that is contiguous with the synaptic vesicle membrane and with the presynaptic plasma membrane and its dense material. In the postsynaptic process, the postsynaptic density (PSD) is seen in intimate contact with the postsynaptic membrane. En face images of the PSD in some synapses appear as a torus. Emanating from the filamentous web of the PSD are filaments which extend to the adjacent plasma membrane. We conclude that membranous synaptic elements are contiguous with a three-dimensional lattice network that is similar to that described in whole unembedded cells (Wolosewick and Porter, 1976). Moreover, the synaptic densities represent a specialized elaboration of the cytoskeleton.


Dietary fat intake and risk of pancreatic cancer in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial.
Epidemiologic and experimental studies suggest that dietary fat intake may affect risk of pancreatic cancer, but published results are inconsistent. We examined risk associations for specific types of dietary fat intakes and related food sources among 111,416 participants in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. We used Cox proportional hazards regression to examine associations between fat intake and pancreatic cancer risk. Over a mean 8.4 years of follow-up, 411 pancreatic cancer cases were identified. We observed an inverse association between saturated fat intake and pancreatic cancer risk (hazard ratio [HR], 0.64 comparing extreme quintiles; 95% confidence interval [CI], 0.46-0.88), but the association became weaker and nonsignificant when individuals with fewer than 4 years of follow-up were excluded to avoid possible reverse causation (HR, 0.88; 95% CI, 0.58-1.33). Total fat intake showed a similar pattern of association, whereas intakes of monounsaturated and polyunsaturated fats and fats from animal or plant sources showed no associations with risk. These results do not support the hypothesis of increased pancreatic cancer risk with higher fat consumption overall or by specific fat type or source. Dietary changes owing to undetected disease may explain the observed inverse association with saturated fat.


[Synthesis of peptide fragments of influenza virus A/Aichi/2/68 (H3N2) hemagglutinins].
Peptides corresponding to sequences 122-133, 136-147, and 154-164 of the heavy chain of hemagglutinin of the A/Aichi/2/68 (H3N2) influenza virus have been synthesized by stepwise elongation of the peptide chain with Boc-amino acid activated esters or by condensation of peptide blocks by DCC/HOBt-method. A coloured C-protecting group, 2-[4-(phenylazo)-benzylsulfonyl]ethyl (PSE), was used, which is convenient in purification of synthetic peptides. After removal of terminal N-and C-protecting groups the side-protecting residues were cleaved off with 1 M trifluoromethanesulfonic acid in trifluoroacetic acid containing 10% thioanisole. Crude products were purified by preparative reversed-phase liquid chromatography. Synthesized peptides were conjugated with BSA.


Prevalent Gaps in Understanding the Features of Catatonia Among Psychiatrists, Psychiatry Trainees, and Medical Students.
Background: Catatonia is often overlooked, and a key factor for underdiagnosis may be an inadequate understanding of catatonia's heterogeneous phenotypes. The aim of this study was to identify the current state of theoretical and applied knowledge of catatonic features among psychiatry trainees and practitioners using the Bush-Francis Catatonia Rating Scale (BFCRS), the most commonly used instrument to identify and score catatonia. Methods: We created an online 50-item multiple-choice test and 3-minute standardized patient video to be scored using the BFCRS. Email invitations were sent to medical students and psychiatry residents and fellows through listservs of psychiatry clerkship and residency directors and to consultation-liaison psychiatrists through the Academy of Consultation-Liaison Psychiatry. Participants could access the exam from October 1 to December 31, 2020. Results: In our sample (n = 482), participants correctly answered an average of 55% of test questions and identified 69% of BFCRS items on the standardized patient exam. Multivariable regression adjusting for demographics revealed that, compared to medical students, psychiatrists scored 7 points higher on the multiple-choice test and identified only 2 more items correctly on the BFCRS. Older participants performed worse than younger participants. No meaningful performance differences were identified by region or gender. Several items were consistently misidentified. Conclusions: We found significant inaccuracies in clinicians' understanding of catatonic features irrespective of their stage of training and years of experience. These data suggest prevalent gaps in catatonia recognition among psychiatrists, psychiatry trainees, and medical students utilizing the BFCRS. This has important implications for clinical research and patient care.


[The future of clinical pathology].
Predictions of the future of pathology must encompass not only anticipation of scientific advances, but the important relationships with other physicians, scientists, health care agencies, hospitals, the public, and the government. The last fifty years have seen advances dwarfing all medical science throughout previous time, with sweeping changes in laboratory medicine in more recent years. For example, advent of molecular pathology will generate opportunities for the pathologist to lead not only in diagnosis, but to accurately predict and prevent disease, and to participate in treatment. Genetic engineering, which promises revolutionary improvement in medicine with already exciting advances in heritable disease. Immune status abnormalities, organ transplantation, and even mental disease, offers great opportunities for clinical pathologists. Currently there is much variation in definition and practice of anatomic and clinical pathology throughout the world. Scientific advances will merge the roles of these two branches more closely. The American Board of Pathology already foresees changing the current separate certifications in anatomic and clinical pathology into one certification in pathology. Multiple subcertifications will be offered to meet the certain increase in applicable clinical science in many related disciplines. The pathologist expert in a new field will assume a more activist role in clinical medicine and will be positioned to become the "information specialist" who will be required, with the aid of computerization, to assemble, correlate, and apply this burgeoning medical knowledge, which in many cases will transcend the abilities of any one medical specialist or allied scientist. The public become insatiable for knowledge and then access to this new medical science.(ABSTRACT TRUNCATED AT 250 WORDS)


Histone deacetylase (HDAC) inhibitors attenuate cardiac hypertrophy by suppressing autophagy.
Histone deacetylases (HDACs) regulate cardiac plasticity; however, their molecular targets are unknown. As autophagy contributes to pathological cardiac remodeling, we hypothesized that HDAC inhibitors target autophagy. The prototypical HDAC inhibitor (HDACi), trichostatin A (TSA), attenuated both load- and agonist-induced hypertrophic growth and abolished the associated activation of autophagy. Phenylephrine (PE)-triggered hypertrophy and autophagy in cultured cardiomyocytes were each blocked by a panel of structurally distinct HDAC inhibitors. RNAi-mediated knockdown of either Atg5 or Beclin 1, two essential autophagy effectors, was similarly capable of suppressing ligand-induced autophagy and myocyte growth. RNAi experiments uncovered the class I isoforms HDAC1 and HDAC2 as required for the autophagic response. To test the functional requirement of autophagic activation, we studied mice that overexpress Beclin 1 in cardiomyocytes. In these animals with a fourfold amplified autophagic response to TAC, TSA abolished TAC-induced increases in autophagy and blunted load-induced hypertrophy. Finally, we subjected animals with preexisting hypertrophy to HDACi, finding that ventricular mass reverted to near-normal levels and ventricular function normalized completely. Together, these data implicate autophagy as an obligatory element in pathological cardiac remodeling and point to HDAC1/2 as required effectors. Also, these data reveal autophagy as a previously unknown target of HDAC inhibitor therapy.


Monitoring methods for local nasal immunotherapy.
When a patient is affected by specific vasomotor rhinopathy (allergic rhinitis), both perennial and seasonal, all the nasal functions are altered to various degrees. For this reason, in the evaluation of the efficacy of an antiallergic therapy we include: 1) anterior rhinoscopy; 2) active anterior rhinomanometry; 3) positioned acoustic rhinometry; 4) determination of mucociliary transport time; and 5) specific nasal provocation test. Active anterior rhinomanometry performed according to the directions of the International Committee on Standardization allows reliable assessment of the nasal respiratory function. Acoustic rhinometry is an objective method to assess the geometry of nasal cavities but, as a new method, needs more standardization with particular care in connecting the nose-piece to the nostril. Both these tests performed before and after local administration of nasal decongestants allow quantification of the congestion degree of the nasal mucosa. Determination of mucociliary transport time, according to the method of coloured tracer, is easy to perform and may be considered a reliable indicator of the mucosa eutrophism. In our clinical experience we have often found mucociliary transport times delayed or even blocked, especially in patients affected by perennial allergic rhinitis. According to our experience the specific nasal provocation test is one of the most important tests in this field: it is more sensitive than skin tests and RAST in the asymptomatic phase and it is able to show organ allergies. In this paper we review the importance of this test, the methodology we currently use as well as changes after local nasal immunotherapy.


Hydrogen Sulfide Alleviates Cadmium-Induced Cell Death through Restraining ROS Accumulation in Roots of Brassica rapa L. ssp. pekinensis.
Hydrogen sulfide (H2S) is a cell signal molecule produced endogenously and involved in regulation of tolerance to biotic and abiotic stress in plants. In this work, we used molecular biology, physiology, and histochemical methods to investigate the effects of H2S on cadmium- (Cd-) induced cell death in Chinese cabbage roots. Cd stress stimulated a rapid increase of endogenous H2S in roots. Additionally, root length was closely related to the cell death rate. Pretreatment with sodium hydrosulfide (NaHS), a H2S donor, alleviated the growth inhibition caused by Cd in roots-this effect was more pronounced at 5 μM NaHS. Cd-induced cell death in roots was significantly reduced by 5 μM NaHS treatment. Under Cd stress, activities of the antioxidant enzymes were significantly enhanced in roots. NaHS + Cd treatment made their activities increase further compared with Cd exposure alone. Enhanced antioxidant enzyme activity led to a decline in reactive oxygen species accumulation and lipid peroxidation. In contrast, these effects were reversed by hydroxylamine, a H2S inhibitor. These results suggested that H2S alleviated the cell death caused by Cd via upregulation of antioxidant enzyme activities to remove excessive reactive oxygen species and reduce cell oxidative damage.


Effects of hypercalcemia and parathyroid hormone on blood pressure in normal and renal-failure rats.
Hypertension is common in primary hyperparathyroidism, but the mechanisms are not clear. Significant hypercalcemia induces elevation in blood pressure (BP), whereas excessive parathyroid hormone (PTH) lowers BP. However, in chronic renal failure (CRF) and secondary hyperparathyroidism, the hypercalcemia-induced hypertension is more severe. We examined the interaction between PTH and calcium on BP in normal rats and in those with CRF. Calcium caused a dose-related rise in serum calcium and a rise in mean arterial pressure (MAP). For a comparable rise in serum calcium, the increment in MAP in parathyroidectomized (PTX) rats (7 +/- 3 mmHg) was significantly lower (P less than 0.05) than in sham PTX rats (19 +/- 7.3 mmHg). In PTX rats receiving PTH, the MAP response to calcium infusion (17 +/- 2.4 mmHg) was similar to that in the sham PTX rats. The infusion of similar amounts of calcium in CRF rats caused a greater rise in serum calcium. In CRF-PTX rats, the changes in MAP during calcium infusion were significantly lower (P less than 0.05) than in CRF-sham PTX animals, despite similar rise in serum calcium. For a comparable rise in serum calcium, the rise in MAP in CRF rats was greater than in normal rats. These data suggest that the presence of PTH plays an important permissive role for the hypertensive action of the hypercalcemia.


Results of Schirmer tear test in clinically normal llamas (Lama glama).
To determine the normal reference range for Schirmer tear test (STT) values in clinically normal llamas (Lama glama) Nine captive llamas (Lama glama) (seven females and two males) were used in this study. Complete ophthalmic examinations were performed without chemical restraint. STT I values were evaluated in both eyes of all llamas using a commercial STT strip of a single lot number (Schirmer-Tränentest(®), Germany). STT II value was also measured in both eyes of seven female llamas. No statistically significant differences among ages or between right and left eyes were found for any of the results. The mean ± SD STT I of 18 eyes of nine llamas was 17.3 ± 1.1 mm/min (Range 15-19 mm/min). The mean ± SD STT II of 14 eyes of seven llamas was 15.4 ± 1.7 mm/min (Range 12.5-17.5 mm/min). A paired samples t-test demonstrated that there was a significant difference between the STT I and II values (P = 0.001). This study provides novel data for normal reference ranges of STT I and II values in healthy llamas. Results of this study may assist veterinarians in the diagnosis of ocular surface disease and syndromes affecting the tear film in these species.


Neuroendocrine tumors of the bronchopulmonary system (typical and atypical carcinoid tumors): current strategies in diagnosis and treatment. Conclusions of an expert meeting February 2011 in Weimar, Germany.
Neuroendocrine tumors (NETs; syn. carcinoid tumors) are highly or moderately differentiated neoplasms. They comprise a large variety of rare and heterogeneous tumors with an estimated incidence of 3-5/100,000/year. They can arise in virtually every internal organ, but mainly occur in the gastroenteropancreatic and bronchopulmonary systems. Around 25% of the NETs are localized in the bronchopulmonary system. Approximately 2% of all lung tumors are NETs. According to the World Health Organization (WHO) classification of lung tumors, bronchopulmonary NETs are subdivided into typical carcinoids (TCs) and atypical carcinoids (ACs). The parameter with the highest impact on NET behavior and prognosis is the histological classification and staging according to the tumor/node/metastasis (TNM) system. The diagnosis of NETs is established by histological examination and the immunohistochemical detection of general neuroendocrine markers, such as chromogranin A (CgA) and synaptophysin. Serum markers and the use of functional imaging techniques are important additive tools to establish the diagnosis of a NET. The only curative option for lung NETs is complete surgical resection. Beyond that, the currently available interdisciplinary therapeutic options are local ablation, biotherapy (somatostatin analogues), or chemotherapy. New therapeutic options such as peptide receptor radionuclide therapy (PRRT) and molecularly targeted therapies achieve promising results and are under further evaluation. This report is a consensus summary of the interdisciplinary symposium 'Neuroendocrine Tumors of the Lung and of the Gastroenteropancreatic System (GEP NET) - Expert Dialogue' held on February 25-26, 2011 in Weimar, Germany. At this conference, a panel of 23 German experts shared their knowledge and exchanged their thoughts about research, diagnosis, and clinical management of NETs, whereby special attention was paid to NETs of the respiratory tract.


Peripheral and central immune cell reservoirs in tissues from asymptomatic cats chronically infected with feline immunodeficiency virus.
Feline immunodeficiency virus (FIV) infection in cats results in life-long viral persistence and progressive immunopathology. We have previously described a cohort of experimentally infected cats demonstrating a progressive decline of peripheral blood CD4+ T-cell over six years in the face of apparent peripheral viral latency. More recently we reported findings from this same cohort that revealed popliteal lymph node tissue as sites for ongoing viral replication suggesting that tissue reservoirs are important in FIV immunopathogenesis during the late asymptomatic phase of infection. Results reported herein characterize important tissue reservoirs of active viral replication during the late asymptomatic phase by examining biopsied specimens of spleen, mesenteric lymph node (MLN), and intestine from FIV-infected and uninfected control cats. Peripheral blood collected coincident with harvest of tissues demonstrated severe CD4+ T-cell depletion, undetectable plasma viral gag RNA and rarely detectable peripheral blood mononuclear cell (PBMC)-associated viral RNA (vRNA) by real-time PCR. However, vRNA was detectable in all three tissue sites from three of four FIV-infected cats despite the absence of detectable vRNA in plasma. A novel in situ hybridization assay identified B cell lymphoid follicular domains as microanatomical foci of ongoing FIV replication. Additionally, we demonstrated that CD4+ leukocyte depletion in tissues, and CD4+ and CD21+ leukocytes as important cellular reservoirs of ongoing replication. These findings revealed that tissue reservoirs support foci of ongoing viral replication, in spite of highly restricted viral replication in blood. Lentiviral eradication strategies will need address tissue viral reservoirs.


A multi-locus species phylogeny of African forest duikers in the subfamily Cephalophinae: evidence for a recent radiation in the Pleistocene.
Duikers in the subfamily Cephalophinae are a group of tropical forest mammals believed to have first originated during the late Miocene. However, knowledge of phylogenetic relationships, pattern and timing of their subsequent radiation is poorly understood. Here we present the first multi-locus phylogeny of this threatened group of tropical artiodactyls and use a Bayesian uncorrelated molecular clock to estimate divergence times. A total of 4152 bp of sequence data was obtained from two mitochondrial genes and four nuclear introns. Phylogenies were estimated using maximum parsimony, maximum likelihood, and Bayesian analysis of concatenated mitochondrial, nuclear and combined datasets. A relaxed molecular clock with two fossil calibration points was used to estimate divergence times. The first was based on the age of the split between the two oldest subfamilies within the Bovidae whereas the second was based on the earliest known fossil appearance of the Cephalophinae and molecular divergence time estimates for the oldest lineages within this group. Findings indicate strong support for four major lineages within the subfamily, all of which date to the late Miocene/early Pliocene. The first of these to diverge was the dwarf duiker genus Philantomba, followed by the giant, eastern and western red duiker lineages, all within the genus Cephalophus. While these results uphold the recognition of Philantomba, they do not support the monotypic savanna-specialist genus Sylvicapra, which as sister to the giant duikers leaves Cephalophus paraphyletic. BEAST analyses indicate that most sister species pairs originated during the Pleistocene, suggesting that repeated glacial cycling may have played an important role in the recent diversification of this group. Furthermore, several red duiker sister species pairs appear to be either paraphyletic (C.callipygus/C. ogilbyi and C. harveyi/C. natalensis) or exhibit evidence of mitochondrial admixture (C. nigrifrons and C. rufilatus), consistent with their recent divergence and/or possible hybridization with each other. Molecular phylogenetic analyses suggest that Pleistocene-era climatic oscillations have played an important role in the speciation of this largely forest-dwelling group. Our results also reveal the most well supported species phylogeny for the subfamily to date, but also highlight several areas of inconsistency between our current understanding of duiker taxonomy and the evolutionary relationships depicted here. These findings may therefore prove particularly relevant to future conservation efforts, given that many species are presently regulated under the Convention for Trade in Endangered Species.


Evolution of the continents and the atmosphere inferred from Th-U-Nb systematics of the depleted mantle
Temporal evolution of depleted mantle thorium-uranium-niobium systematics constrain the amount of continental crust present through Earth's history (through the niobium/thorium ratio) and date formation of a globally oxidizing atmosphere and hydrosphere at approximately 2.0 billion years ago (through the niobium/uranium ratio). Increase in the niobium/thorium ratio shows involvement of hydrated lithosphere in differentiation of Earth since approximately 3.8 billion years ago. After approximately 2.0 billion years ago, the decreasing mantle thorium/uranium ratio portrays mainly preferential recycling of uranium in an oxidizing atmosphere and hydrosphere. Net growth rate of continental crust has varied over time, and continents are still growing today.


Deficiency of a new protein associated with cardiac syndrome X; called adropin.
The pathophysiology of cardiac syndrome X (CSX) is still unclear, but most patients with CSX have endothelial dysfunction. It has been shown that adropin uniquely effects the regulation of endothelial function. The purpose of the study was to evaluate the role of adropin in CSX. Eighty-six consecutive cardiac syndrome X-diagnosed patients and 86 age-sex matched healthy subjects were enrolled into the study. Serum adropin levels, nitrite/nitrate levels were measured in each subject. The adropin levels were significantly lower in patients with CSX than healthy subjects (1.7 ± 0.8 ng/mL and 3.4 ± 1.8 ng/mL, respectively; P < 0.001). The BMI values of patients with CSX were significantly higher than control subjects (28.1 ± 2.4 kg/m(2) and 26.0 ± 3.7 kg/m(2) , respectively; P < 0.001). Plasma nitrite/nitrate levels were lower in patients with CSX than control subjects (15.9 ± 1.6 μmol/L vs. 25.4 ± 2.8 μmol/L, respectively; P < 0.001), and they have a significantly positive correlation with plasma adropin levels (r = 0.463, P < 0.001). In the multiple linear regression analysis, nitrite/nitrate levels, BMI, and adropin were found to be independent risk factors for CSX. A ROC curve is used to identify the ability of adropin levels to predict the cardiac syndrome X. The area under the ROC curve was 0.854 for adropin levels (P = 0.0001). The sensitivity and specificity values of adropin levels were 90.7 and 70.9%, respectively (cut-off value 2.73). In conclusion, lower serum adropin levels were associated with CSX. Adropin is an independent risk factor for CSX.


Tobacco smoking and bipolar disorder.
We sought to determine whether tobacco smoking is associated with bipolar disorder. This case-control study carried out in Alava, in the north of Spain, included patients with a DSM-III-R diagnosis of bipolar disorder type I (N = 51) and a representative sample of the normal population (N = 517). Smoking history of bipolar patients was assessed with the Fagerstrom Test for Nicotine Dependence and was verified by family members of the patients. The frequencies of ever smoking and current daily smoking were, respectively, 63% (32/51) and 51% (26/51) for the bipolar patients and 45% (235/517) and 33% (169/517) for the control group (respective odds ratios [ORs] and 95% confidence intervals [CIs] were OR = 2.0, 95% CI = 1.1 to 3.8 and OR = 2.1, 95% CI = 1.2 to 4.0). The differences were significant (p = .03 and p = .042, respectively) for bipolar versus control males. Bipolar disorder (in both genders) was also significantly associated (OR = 4.4, 95% CI = 1.7 to 11.9, p = .0015) with heavy smoking (more than 1 pack per day). Despite its small sample size, this study suggests that smoking may be more prevalent in bipolar patients than in the normal population. Since most patients started to smoke before the onset of illness, vulnerability to bipolar illness may make subjects vulnerable to become tobacco smokers.


Characteristics of patients presenting with ocular pain.
The purpose of this study was to assess the utility of neuroimaging in patients with normal neuro-ophthalmic examinations referred to a neuro-ophthalmology clinic for complaints of chronic isolated eye pain. We were also interested in the prevalence of primary headache syndromes in these patients, particularly medication overuse headache (MOH). Charts of 125 consecutive patients referred to a neuro-ophthalmology clinic with some form of head or facial pain were reviewed to find patients meeting the criteria of having complaints of chronic eye with normal neuro-ophthalmic examinations. Excluded were patients with primary complaint other than eye pain. A single examiner performed all examinations in a standardized fashion. Each patient was queried about the use of prescription and over-the-counter analgesics. Seventy-three percent of patients had some form of primary headache syndrome, and 21% of patients had some component of probable MOH. Another 4.5% had a presumed component of medication overuse. None of those who received imaging had any results explaining the ocular pain. Primary headache is present in most patients with chronic, isolated ocular pain. MOH is common and probably underrecognized in this population. It needs to be addressed if the patient is ever to be pain free. Neuroimaging has very low yield in this group of subspecialty referral patients.


Migraine as a Disease Associated with Dysbiosis and Possible Therapy with Fecal Microbiota Transplantation.
Migraine is a painful neurological condition characterized by severe pain on one or both sides of the head. It may be linked to changes in the gut microbiota, which are influenced by antibiotic use and other factors. Dysbiosis, which develops and persists as a result of earlier antibiotic therapy, changes the composition of the intestinal flora, and can lead to the development of various diseases such as metabolic disorders, obesity, hematological malignancies, neurological or behavioral disorders, and migraine. Metabolites produced by the gut microbiome have been shown to influence the gut-brain axis. The use of probiotics as a dietary supplement may reduce the number and severity of migraine episodes. Dietary strategies can affect the course of migraines and are a valuable tool for improving migraine management. With fecal microbiota transplantation, gut microbial restoration is more effective and more durable. Changes after fecal microbiota transplantation were studied in detail, and many data help us to interpret the successful interventions. The microbiological alteration of the gut microflora can lead to normalization of the inflammatory mediators, the serotonin pathway, and influence the frequency and intensity of migraine pain.


Probiotics, prebiotics, and synbiotics: impact on the gut immune system and allergic reactions.
Probiotics and prebiotics, alone or together (synbiotics), can influence the intestinal microbiota and modulate the immune response. They may therefore be tools that can prevent or alleviate certain pathologies involving the gut immune system, such as allergies for which no treatment is yet available. This review focuses first on the definitions of probiotics, prebiotics, and synbiotics and key cells in the gut immune system. It then discusses their effects on mucosal immune stimulation. Experimental findings suggest that different probiotic species have similar effects on innate immunity by improving the mechanisms of pathogen destruction. On the contrary, their impacts seem to be variable on the adaptive immune system. Prebiotics can also exert an influence on the gut immune system via the stimulation of the autochthonous bacteria metabolism. Finally, this review focuses on the effects of food supplements on allergy. Different studies performed in humans or rodents have supported a potential role for selected probiotics and prebiotics in reducing some allergic parameters. Probiotic effects on allergy treatment are unclear, especially in human studies. However, they are potentially effective at short-term for prevention when they are administered in perinatal conditions. A clinical study performed with an infant cohort revealed a beneficial effect of prebiotics in preventing allergic manifestations at long-term. Further studies are nonetheless essential to confirm these findings. Food supplements offer potential tools for the prevention or treatment of allergy, but insufficient evidence is available at present to recommend their use in clinical practice.


[The role of microsporidia in the disturbance of the hormonal balance in the infected insects].
Since investigations of Fisher and Sanborn (1964), the most popular point of view on pathological alterations in the development of insects is the statement that microsporidians can produce and secrete the insect juvenile hormone in infected tissues. We suppose that the juvenilizing effect of microsoporidian infection may be a consequence of stress induced by the protozoan invasion. To our opinion, microsporidians can not be a source of juvenile hormone in parasitized insects because of several reasons. 1. The juvenilizing effect of infection may be explained by other reason, that is the stress effect of invasion. 2. Until now nobody can find juvenile hormone in microsporidians. 3. An increase of juvenile hormone titre either alters non-metabolic activity of fat body or alters it in a way that is not favorable for microsporidias. 4. Some effects of the microsporidian infection more resemble an action of ecdysone, but not juvenile hormone. Hypothesis of parasitic stress can explain this, while hypothesis of production JH by microsporidians can not. 5. An effect of invasion by parasites of other systematic groups onto the hormone balance is of the same type. But nobody thinks that other parasites can produce juvenile hormone. Nevertheless it is clear that a high titre of the juvenile hormone is favorable for the microsporidian infection in larvae, especially the last instars. The most descriptions of microsporidian infections are related with this age of insects. Juvenilizing effect of Nosema implantant in the work of Fisher and Sanborn (1964) may be easily explained by decreasing of JH-esterase activity. But in this case the decrease of JH-esterase activity after the implantation of infected tissue in a healthy insect should be explained. We suppose that this fact witness the existence of stress-factor produced by damaged cells of fat body that can enter the healthy cells of fat body and inhibit the activity of JH-esterases.


Extended safety, immunogenicity and efficacy of a blood-stage malaria vaccine in malian children: 24-month follow-up of a randomized, double-blinded phase 2 trial.
The FMP2.1/AS02A candidate malaria vaccine was tested in a Phase 2 study in Mali. Based on results from the first eight months of follow-up, the vaccine appeared well-tolerated and immunogenic. It had no significant efficacy based on the primary endpoint, clinical malaria, but marginal efficacy against clinical malaria in secondary analyses, and high allele-specific efficacy. Extended follow-up was conducted to evaluate extended safety, immunogenicity and efficacy. A randomized, double-blinded trial of safety, immunogenicity and efficacy of the candidate Plasmodium falciparum apical membrane antigen 1 (AMA1) vaccine FMP2.1/AS02A was conducted in Bandiagara, Mali. Children aged 1-6 years were randomized in a 1∶1 ratio to receive FMP2.1/AS02A or control rabies vaccine on days 0, 30 and 60. Using active and passive surveillance, clinical malaria and adverse events as well as antibodies against P. falciparum AMA1 were monitored for 24 months after the first vaccination, spanning two malaria seasons. 400 children were enrolled. Serious adverse events occurred in nine participants in the FMP2.1/AS02A group and three in the control group; none was considered related to study vaccination. After two years, anti-AMA1 immune responses remained significantly higher in the FMP2.1/AS02A group than in the control group. For the entire 24-month follow-up period, vaccine efficacy was 7.6% (p = 0.51) against first clinical malaria episodes and 9.9% (p = 0.19) against all malaria episodes. For the final 16-month follow-up period, vaccine efficacy was 0.9% (p = 0.98) against all malaria episodes. Allele-specific efficacy seen in the first malaria season did not extend into the second season of follow-up. Allele-specific vaccine efficacy was not sustained in the second malaria season, despite continued high levels of anti-AMA1 antibodies. This study presents an opportunity to evaluate correlates of partial protection against clinical malaria that waned during the second malaria season. Clinicaltrials.gov NCT00460525 NCT00460525.


The new Childhood Arthritis and Rheumatology Research Alliance (CARRA) registry: design, rationale, and characteristics of patients enrolled in the first 12 months.
Herein we describe the history, design, and rationale of the new Childhood Arthritis and Rheumatology Research Alliance (CARRA) Registry and present the characteristics of patients with juvenile idiopathic arthritis (JIA) enrolled in the first 12 months of operation. The CARRA Registry began prospectively collecting data in the United States and Canada in July 2015 to evaluate the safety of therapeutic agents in persons with childhood-onset rheumatic disease, initially restricted to JIA. Secondary objectives include the evaluation of disease outcomes and their associations with medication use and other factors. Data are collected every 6 months and include clinical assessments, detailed medication use, patient-reported outcomes, and safety events. Follow-up is planned for at least 10 years for each participant and is facilitated by a telephone call center. As of July 2016, 1192 patients with JIA were enrolled in the CARRA Registry at 49 clinical sites. At enrollment, their median age was 12.4 years old and median disease duration was 2.6 years. Owing to preferential enrollment, patients with systemic JIA (13%) and with a polyarticular course (75%) were over-represented compared to patients in typical clinical practice. Approximately 49% were currently using biologic agents and ever use of oral glucocorticoids was common (47%). The CARRA Registry provides safety surveillance data to pharmaceutical companies to satisfy their regulatory requirements, and several independently-funded sub-studies that use the Registry infrastructure are underway. The new CARRA Registry successfully enrolled nearly 1200 participants with JIA in the first 12 months of its operation. Sustainable funding has been secured from multiple sources. The CARRA Registry may serve as a model for the study of other uncommon diseases.


Oxidative stress-induced depolymerization of microtubules and alteration of mitochondrial mass in human cells.
Mitochondrial biogenesis is a biological process that has been intensively studied over the past few years. However, the detailed molecular mechanism underlying this increase in mitochondria remains unclear. To investigate the mechanism of such a mitochondrial proliferation, we examined alterations in mitochondria of human osteosarcoma 143B cells that had been treated with 100 to 500 microM hydrogen peroxide (H2O2) for 48 h. The results showed that mitochondrial mass of the cell was increased with the increase of the concentration of H2O2. On the other hand, by using real-time PCR techniques, we observed the changes of mitochondrial DNA (mtDNA) content in the cells exposed to oxidative stress. The copy number of mtDNA was increased by treatment with a low dose of H2O2 but was drastically decreased after treatment with H2O2 higher than 300 microM. Transmission electron microscopic images revealed that mitochondria were abnormally proliferated in cells exposed to oxidative stress. Moreover, we found that the percentage of 143B cells arrested at the G2/M phase increased upon treatment with H2O2. Immunostaining and microtubule fractionation assay revealed that microtubules were depolymerized in the cells that had been treated with H2O2. To understand the effect of microtubules depolymerization on the mitochondrial mass, we treated the cells with several kinds of microtubule-active drugs, which arrest cultured cells at the G2/M phase. The results showed that mitochondrial mass and mtDNA copy number all were increased after such treatments. Taking these findings together, we suggest that oxidative stress-induced microtubule derangement is one of the molecular events involved in the increase of mitochondrial mass upon treatment of human cells with H2O2.