Tissue-engineered nipple reconstruction.
We describe a simple, effective approach to the creation of autologous tissue-engineered cartilage in the shape of a human nipple by injecting a reverse thermosensitive polymer seeded with autologous chondrocytes in an immunocompetent porcine animal model. A biodegradable, biocompatible copolymer of polyethylene oxide and polypropylene oxide (Pluronic F-127), which exists as a liquid below 4 degrees C and polymerizes to a thick gel when it is exposed to physiologic temperatures (body temperatures), was used as a vehicle for chondrocyte delivery and as a scaffold to guide growth. Autologous chondrocytes isolated from porcine auricular elastic cartilage and suspended in 30% (weight/volume) Pluronic F-127 were injected on the ventral surface of the pigs from which the cells had been isolated. A circumferential subdermal suture was used to support the contour of the implant and assist in its projection in the form of a human nipple. After 3 weeks, the skin over and surrounding the implant was tattooed to create the appearance of a human nipple-areolar complex. As controls, an equal number of injections were made using either cells alone (not suspended in hydrogel), or hydrogel alone. After 10 weeks, all specimens were excised and examined both grossly and histologically. Before harvesting, visual inspection of the tattooed chondrocyte-Pluronic F-127 hydrogel implant sites revealed that they closely resembled a human female nipple-areolar complex. Nodules were similar in size, shape, and texture to a human nipple at each injection site. Glistening opalescent tissue was surgically isolated from each implant site. Hematoxylin and eosin, safranine o, trichrome blue, and Verhoeff's stains of the experimental implants showed nodules with the characteristic histologic signs of elastic cartilage. Control injections of copolymer hydrogel alone exhibited no evidence of cartilage formation. Control injections of chondrocytes alone showed evidence of dissociated microscopic nodules of elastic cartilage.


Wound care for persons with diabetes.
This article addresses local wound care in people with diabetes mellitus. An approach is proposed that uses wound depth and color to determine the goals of local wound treatment. Principles of cleansing, debridement, and the use of moisture retentive dressings are discussed. Infection, a significant problem in the diabetic patient, is addressed as an essential part of local wound care in these patients.


Conductance-Based Adaptive Exponential Integrate-and-Fire Model.
The intrinsic electrophysiological properties of single neurons can be described by a broad spectrum of models, from realistic Hodgkin-Huxley-type models with numerous detailed mechanisms to the phenomenological models. The adaptive exponential integrate-and-fire (AdEx) model has emerged as a convenient middle-ground model. With a low computational cost but keeping biophysical interpretation of the parameters, it has been extensively used for simulations of large neural networks. However, because of its current-based adaptation, it can generate unrealistic behaviors. We show the limitations of the AdEx model, and to avoid them, we introduce the conductance-based adaptive exponential integrate-and-fire model (CAdEx). We give an analysis of the dynamics of the CAdEx model and show the variety of firing patterns it can produce. We propose the CAdEx model as a richer alternative to perform network simulations with simplified models reproducing neuronal intrinsic properties.


Intestinal microbiota research from a global perspective.
The intestinal microbiota plays a crucial role in health and changes in its composition are linked with major global human diseases. Fully understanding what shapes the human intestinal microbiota composition and knowing ways of modulating the composition are critical for promotion of life-course health, combating diseases, and reducing global health disparities. We aim to provide a foundation for understanding what shapes the human intestinal microbiota on an individual and global scale, and how interventions could utilize this information to promote life-course health and reduce global health disparities. We briefly review experiences within the first 1,000 days of life and how long-term exposures to environmental elements or geographic specific cultures have lasting impacts on the intestinal microbiota. We also discuss major public health threats linked to the intestinal microbiota, including antimicrobial resistance and disappearing microbial diversity due to globalization. In order to promote global health, we argue that the interplay of the larger ecosystem with intestinal microbiota research should be utilized for future research and urge for global efforts to conserve microbial diversity.


Autophagic Inhibition via Lysosomal Integrity Dysfunction Leads to Antitumor Activity in Glioma Treatment.
Manipulating autophagy is a promising strategy for treating cancer as several autophagy inhibitors are shown to induce autophagic cell death. One of these, autophagonizer (APZ), induces apoptosis-independent cell death by binding an unknown target via an unknown mechanism. To identify APZ targets, we used a label-free drug affinity responsive target stability (DARTS) approach with a liquid chromatography/tandem mass spectrometry (LC-MS/MS) readout. Of 35 protein interactors, we identified Hsp70 as a key target protein of unmodified APZ in autophagy. Either APZ treatment or Hsp70 inhibition attenuates integrity of lysosomes, which leads to autophagic cell death exhibiting an excellent synergism with a clinical drug, temozolomide, in vitro, in vivo, and orthotropic glioma xenograft model. These findings demonstrate the potential of APZ to induce autophagic cell death and its development to combinational chemotherapeutic agent for glioma treatment. Collectively, our study demonstrated that APZ, a new autophagy inhibitor, can be used as a potent antitumor drug candidate to get over unassailable glioma and revealed a novel function of Hsp70 in lysosomal integrity regulation of autophagy.


Differences in Cervical Cancer Screening Knowledge and Practices by HIV Status and Geographic Location: Implication for Program Implementation in Zambia.
The knowledge and perceptions of cervical cancer among HIV negative and positive women, aged 25-49 years, from rural and urban locations in Zambia was systematically accessed in this study to determine any differences. Data were coded and analyzed using NVivo software. Compared to HIV negative women, HIV positive women had more accurate information about cervical cancer. They were more likely to cite male circumcision as the best approach to cervical cancer prevention. HPV infection was more commonly mentioned as a risk factor among HIV positive women. However, HIV positive women displayed little knowledge about HPV being the major cause of cervical cancer. Among HIV positive women, lack of time was the major screening barrier cited while HIV negative women mentioned being symptomatic as a determinant for early detection. Compared to rural residents, urban residents cited a wider range of cervical cancer information sources, including media and workplace although all of the participants who stated that they had no knowledge of cervical cancer were urban residents. Overall, knowledge and perceptions of cervical cancer among study participants was high, although differences exist between subgroups. Sharing accurate and standardized information on cervical cancer would improve participation in cervical cancer screening services.


Normalization of pre-ASCT, FDG-PET imaging with second-line, non-cross-resistant, chemotherapy programs improves event-free survival in patients with Hodgkin lymphoma.
We previously reported that remission duration < 1 year, extranodal disease, and B symptoms before salvage chemotherapy (SLT) can stratify relapsed or refractory Hodgkin lymphoma (HL) patients into favorable and unfavorable cohorts. In addition, pre-autologous stem cell transplant (ASCT) (18)FDG-PET response to SLT predicts outcome. This phase 2 study uses both pre-SLT prognostic factors and post-SLT FDG-PET response in a risk-adapted approach to improve PFS after high-dose radio-chemotherapy (HDT) and ASCT. The first SLT uses 2 cycles of ICE in a standard or augmented dose (ICE/aICE), followed by restaging FDG-PET scan. Patients with a negative scan received a transplant. If the FDG-PET scan remained positive, patients received 4 biweekly doses of gemcitabine, vinorelbine, and liposomal doxorubicin. Patients without evidence of disease progression proceeded to HDT/ASCT; those with progressive disease were study failures. At a median follow-up of 51 months, EFS analyzed by intent to treat as well as for transplanted patients is 70% and 79%, respectively. Patients transplanted with negative FDG-PET, pre-HDT/ASCT after 1 or 2 SLT programs, had an EFS of > 80%, versus 28.6% for patients with a positive scan (P < .001). This prospective study provides evidence that the goal of SLT in patients with Hodgkin lymphoma should be a negative FDG-PET scan before HDT/ASCT.


A model to facilitate the mental health of psychiatric nurses in a forensic unit to manage mental health care users' hostile behaviour constructively.
Hostile behaviour by mental health care users (MHCUs) is prevalent in forensic units in South Africa, and this causes service providers distress and burnout. Psychiatric nurses (PNs) find it difficult to render quality care to MHCUs who are threatening them and also challenging their authority in a forensic unit. Forensic mental health care practitioners may be challenged to engage authentically with MHCUs who constitute a risk to their personal safety or who have committed acts the practitioner finds morally disturbing. There is a need to facilitate the mental health of PNs in a forensic unit to manage hostile behaviour constructively. The objective of this article is to describe the process that was followed in developing, implementing and evaluating a model that could be used as a framework of reference to facilitate the mental health of PNs in a forensic unit to manage hostile behaviour constructively. A theory-generative, qualitative, exploratory descriptive and contextual study design was used to develop the model. The steps of the process entailed the identification of the central concept, the definition of the central concept and other essential criteria and the classification of the central and related concepts. The model was then described and evaluated. The central concept was identified as the 'facilitation of empowerment of PNs to manage hostility in a constructive manner', defined, classified and then described and evaluated. The model as framework of reference could assist PNs in managing hostility in a forensic unit constructively.


Chiral six-coordinate Dy(iii) and Tb(iii) complexes of an achiral ligand: structure, fluorescence, and magnetism.
Two new chiral six-coordinate lanthanide complexes (1 for Dy and 2 for Tb) have been synthesized using the achiral flexible ligand H2L (2,2'-[1,2-ethanediylbis[(methylimino)methylene]]bis[4,5-dimethylphenol]). Both complexes crystallize in the chiral P1 group space, and the enantiomers [Zn(L)Cl]3Dy·MeOH·0.5H2O (Λ-1), [Zn(L)Cl]3Dy·1.5H2O (Δ-1), [Zn(L)Cl]3Tb·1.5H2O (Λ-2) and [Zn(L)Cl]3Tb·H2O (Δ-2) are isolated in the reaction. Three [Zn(L)Cl]- anions coordinate to the central lanthanide ion using two phenoxo oxygen atoms of L2-, and the lanthanide ion has the coordination geometry of D3. Complex 1 exhibits a field-induced slow magnetization relaxation, which is typical of a single-ion magnet (SIM). Ab initio calculations on complex 1 and studies on magneto-structural correlationship of six-coordinate Dy(iii) SIMs indicate that the small energy barrier of complex 1 might be related to the absence of a unique anisotropic axis for the regular octahedral coordination configuration of Dy(iii) and the high quantum tunneling gap between the ground states (mJ = ±15/2). Complex 2 displays green photofluorescence and triboluminescence.


Advancements in the pathogenesis of hepatic osteodystrophy and the potential therapeutic of mesenchymal stromal cells.
Hepatic osteodystrophy (HOD) is a metabolically associated bone disease mainly manifested as osteoporosis with the characteristic of bone loss induced by chronic liver disease (CLD). Due to its high incidence in CLD patients and increased risk of fracture, the research on HOD has received considerable interest. The specific pathogenesis of HOD has not been fully revealed. While it is widely believed that disturbance of hormone level, abnormal secretion of cytokines and damage of intestinal barrier caused by CLD might jointly affect the bone metabolic balance of bone formation and bone absorption. At present, the treatment of HOD is mainly to alleviate the bone loss by drug treatment, but the efficacy and safety are not satisfactory. Mesenchymal stromal cells (MSCs) are cells with multidirectional differentiation potential, cell transplantation therapy based on MSCs is an emerging therapeutic approach. This review mainly summarized the pathogenesis and treatment of HOD, reviewed the research progress of MSCs therapy and the combination of MSCs and scaffolds in the application of osteoporotic bone defects, and discussed the potential and limitations of MSCs therapy, providing theoretical basis for subsequent studies.


Implementation and Evaluation of a Residency Preparation Program for Fourth-Year Doctor of Pharmacy Students.
To describe the impact of a formal residency preparation program on student match rates, and to evaluate student-reported advisement activities and perceptions of the residency application process. An optional, noncredit-bearing, residency preparation program was implemented in professional year 4 (PY4) of the Doctor of Pharmacy curriculum. The program consisted of 4 residency preparation presentations and/or workshops: curriculum vitae writing, navigating the residency application process and American Society of Health-Systems Pharmacy Midyear Clinical Meeting, letter of intent writing, and interview skills. Students attended either virtually or in person, with 3 of the 4 sessions including small group breakout sessions. The program also included dedicated, 1-on-1 residency advisement with residency-experienced advisors. Residency match rates following program implementation increased from 74.3% (comparison group) to 87.5% (intervention group). More students in the intervention group reported that their advisor assisted them with curriculum vitae review, letter of intent review, and interview skills. In addition, the intervention group reported significantly more time spent meeting with their advisor during PY4 than the comparison group. Students found the program to be beneficial to their professional development, indicated that it helped them to obtain a residency position, and expressed that they would participate in the residency preparation program again. Implementation of a formal residency preparation program for PY4 students that included 1-on-1 dedicated residency advisement increased match rates and interaction between students and their residency advisor.


Dioxin-induced chloracne--reconstructing the cellular and molecular mechanisms of a classic environmental disease.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is among the most toxic pollutants known to date that serves as a prototype for a group of halogenated hydrocarbon compounds characterized by extraordinary environmental persistence and unique ability to concentrate in animal and human tissues. TCDD can elicit a complex array of pleiotropic adverse effects in humans, although chloracne, a specific type of acne-like skin disease, is the only consistent manifestation of dioxin intoxication, thus representing a 'hallmark' of TCDD exposure. Chloracne is considered to be one of the most specific and sensitive biomarkers of TCDD intoxication that allows clinical and epidemiological evaluation of exposure level at threshold doses. The specific cellular and molecular mechanisms involved in pathogenesis of chloracne are still unknown. In this review, we summarize the available clinical data on chloracne and recent progress in understanding the role of the dioxin-dependent pathway in the control of gene transcription and discuss molecular and cellular events potentially involved in chloracne pathogenesis. We propose that the dioxin-induced activation of skin stem cells and a shift in differentiation commitment of their progeny may represent a major mechanism of chloracne development.


Plasma haloperidol levels drawn at neuroleptic threshold doses: a pilot study.
The daily haloperidol doses of 33 newly admitted, acutely psychotic schizophrenic patients were rapidly adjusted to a point at which slight hypokinesia-rigidity first appeared on clinical examination (the neuroleptic threshold). The mean daily haloperidol dose at which the neuroleptic threshold was crossed was 4.2 +/- 2.4 mg/day. The plasma haloperidol levels obtained at these neuroleptic threshold doses clustered around a mean of 4.9 +/- 2.9 ng/ml. This plasma haloperidol level distribution extensively overlaps the proposed lower boundaries for the therapeutically effective range of plasma haloperidol levels, but it is below the range in which coarse extrapyramidal side effects appear in high frequency (greater than 10 ng/ml). Sixty-seven percent of our patients demonstrated moderate or greater therapeutic improvement within 3 weeks of treatment at neuroleptic threshold doses, a therapeutic efficacy rate identical to that expected with standard neuroleptic doses. The authors propose that the neuroleptic threshold is a readily available clinical marker which identifies a range of plasma haloperidol levels capable of producing therapeutic response with minimal associated coarse extrapyramidal side effects.


Functional, structural, and metabolic abnormalities of the hippocampal formation in Williams syndrome.
Williams syndrome (WS), caused by microdeletion of some 21 genes on chromosome 7q11.23, is characterized by dysmorphic features, mental retardation or learning difficulties, elastin arteriopathy, and striking neurocognitive and social-behavioral abnormalities. Recent studies of murine knockouts of key genes in the microdeleted region, LIM kinase 1 (LIMK1) and cytoplasmatic linker protein 2 (CYLN2), demonstrated significant functional and metabolic abnormalities, but grossly normal structure, in the hippocampal formation (HF). Furthermore, deficits in spatial navigation and long-term memory, major cognitive domains dependent on hippocampal function, have been described in WS. We used multimodal neuroimaging to characterize hippocampal structure, function, and metabolic integrity in 12 participants with WS and 12 age-, sex-, and IQ-matched healthy controls. PET and functional MRI studies showed profound reduction in resting blood flow and absent differential response to visual stimuli in the anterior HF in WS. Spectroscopic measures of N-acetyl aspartate, considered a marker of synaptic activity, were reduced. Hippocampal size was preserved, but subtle alterations in shape were present. These data demonstrate abnormalities in HF in WS in agreement with murine models, implicate LIMK1 and CYLN2 in human hippocampal function, and suggest that hippocampal dysfunction may contribute to neurocognitive abnormalities in WS.


Evolutionary systems biology of amino acid biosynthetic cost in yeast.
Every protein has a biosynthetic cost to the cell based on the synthesis of its constituent amino acids. In order to optimise growth and reproduction, natural selection is expected, where possible, to favour the use of proteins whose constituents are cheaper to produce, as reduced biosynthetic cost may confer a fitness advantage to the organism. Quantifying the cost of amino acid biosynthesis presents challenges, since energetic requirements may change across different cellular and environmental conditions. We developed a systems biology approach to estimate the cost of amino acid synthesis based on genome-scale metabolic models and investigated the effects of the cost of amino acid synthesis on Saccharomyces cerevisiae gene expression and protein evolution. First, we used our two new and six previously reported measures of amino acid cost in conjunction with codon usage bias, tRNA gene number and atomic composition to identify which of these factors best predict transcript and protein levels. Second, we compared amino acid cost with rates of amino acid substitution across four species in the genus Saccharomyces. Regardless of which cost measure is used, amino acid biosynthetic cost is weakly associated with transcript and protein levels. In contrast, we find that biosynthetic cost and amino acid substitution rates show a negative correlation, but for only a subset of cost measures. In the economy of the yeast cell, we find that the cost of amino acid synthesis plays a limited role in shaping transcript and protein expression levels compared to that of translational optimisation. Biosynthetic cost does, however, appear to affect rates of amino acid evolution in Saccharomyces, suggesting that expensive amino acids may only be used when they have specific structural or functional roles in protein sequences. However, as there appears to be no single currency to compute the cost of amino acid synthesis across all cellular and environmental conditions, we conclude that a systems approach is necessary to unravel the full effects of amino acid biosynthetic cost in complex biological systems.


Supplementation of infant formula with probiotics and/or prebiotics: a systematic review and comment by the ESPGHAN committee on nutrition.
Infant formulae are increasingly supplemented with probiotics, prebiotics, or synbiotics despite uncertainties regarding their efficacy. The present article, developed by the Committee on Nutrition of the European Society for Paediatric Gastroenterology, Hepatology, and Nutrition, systematically reviews published evidence related to the safety and health effects of the administration of formulae supplemented with probiotics and/or prebiotics compared with unsupplemented formulae. Studies in which probiotics/prebiotics were not administered during the manufacturing process, but thereafter, for example in capsules, the contents of which were supplemented to infant formula or feeds, were excluded.On the basis of this review, available scientific data suggest that the administration of currently evaluated probiotic- and/or prebiotic-supplemented formula to healthy infants does not raise safety concerns with regard to growth and adverse effects. The safety and clinical effects of 1 product should not be extrapolated to other products. At present, there is insufficient data to recommend the routine use of probiotic- and/or prebiotic-supplemented formulae. The Committee considers that the supplementation of formula with probiotics and/or prebiotics is an important field of research. There is a need in this field for well-designed and carefully conducted randomised controlled trials, with relevant inclusion/exclusion criteria and adequate sample sizes. These studies should use validated clinical outcome measures to assess the effects of probiotic and/or prebiotic supplementation of formulae. Such trials should also define the optimal doses and intake durations, as well as provide more information about the long-term safety of probiotics and/or prebiotics. Because most of the trials were company funded, independent trials, preferentially financed jointly by national/governmental/European Union bodies and other international organisations, would be desirable.


Stromal Nerve Imaging and Tracking Using Micro-Optical Coherence Tomography.
To image, track and map the nerve fiber distribution in excised rabbit corneas over the entire stromal thickness using micro-optical coherence tomography (µOCT) to develop a screening tool for early peripheral neuropathy. Excised rabbit corneas were consecutively imaged by a custom-designed µOCT prototype and a commercial laser scanning fluorescence confocal microscope. The µOCT images with a field of view of approximately 1 × 1 mm were recorded with axial and transverse resolutions of approximately 1 µm and approximately 4 µm, respectively. In the volumetric µOCT image data, network maps of hyper-reflective, branched structures traversing different stromal compartments were segmented using semiautomatic image processing algorithms. Furthermore, the same corneas received βIII-tubulin antibody immunostaining before digital confocal microscopy, and a comparison between µOCT image data and immunohistochemistry analysis was performed to validate the nerval origin of the tracked network structures. Semiautomatic tracing of the nerves with a high range of different thicknesses was possible through the whole corneal volumes, creating a skeleton of the traced nerves. There was a good conformity between the hyper-reflective structures in the µOCT data and the stained nerval structures in the immunohistochemistry data. This article demonstrates nerval imaging and tracking as well as a spatial correlation between µOCT and a fluorescence corneal nerve standard for larger nerves throughout the full thickness of the cornea ex vivo. Owing to its advantageous properties, µOCT may become useful as a noncontact method for assessing nerval structures in humans to screen for early peripheral neuropathy.


[Outpatient and inpatient orthopaedic rehabilitation--results of a study comparing outcome and costs].
This article presents main results of a study comparing outcome and costs of various all-day outpatient and inpatient orthopaedic rehabilitation forms. The results were obtained within the scope of a comprehensive evaluation programme commissioned by the federation of health (VdAK) and pension insurance institutes (VDR). The purpose of this evaluation was to examine the quality of different types of various rehabilitation care. The outcome comparison comprises a longitudinal follow-up of various somatic, psychosocial und occupational reintegration parameters up to one year after rehabilitation as well as outcome assessments from the perspective of patients and physicians in rehabilitation centres and general practice. The costs are estimated on the basis of data from health and pension insurance funds. The results show extensive corresponding of results und assessments in the different forms of rehabilitation care for comparable patient groups. There are no indications of poorer care quality in outpatient rehabilitation, while economic analyses show better cost effectiveness in outpatient treatment by comparability of treatment, patients, and results. Results suggest that outpatient care, offered in the same quality as in the examined rehabilitation centres, is an alternative or complement to inpatient care at least for those patients, who can be treated in both the outpatient and inpatient setting.


Generation of transgenic rodent malaria parasites by transfection of cell culture-derived merozoites.
Malaria research is greatly dependent on and has drastically advanced with the possibility of genetically modifying Plasmodium parasites. The commonly used transfection protocol by Janse and colleagues utilizes blood stage-derived Plasmodium berghei schizonts that have been purified from a blood culture by density gradient centrifugation. Naturally, this transfection protocol depends on the availability of suitably infected mice, constituting a time-based variable. In this study, the potential of transfecting liver stage-derived merozoites was explored. In cell culture, upon merozoite development, infected cells detach from the neighbouring cells and can be easily harvested from the cell culture supernatant. This protocol offers robust experimental timing and temporal flexibility. HeLa cells are infected with P. berghei sporozoites to obtain liver stage-derived merozoites, which are harvested from the cell culture supernatant and are transfected using the Amaxa Nucleofector® electroporation technology. Using this protocol, wild type P. berghei ANKA strain and marker-free PbmCherryHsp70-expressing P. berghei parasites were successfully transfected with DNA constructs designed for integration via single- or double-crossover homologous recombination. An alternative protocol for Plasmodium transfection is hereby provided, which uses liver stage-derived P. berghei merozoites for transfection. This protocol has the potential to substantially reduce the number of mice used per transfection, as well as to increase the temporal flexibility and robustness of performing transfections, if mosquitoes are routinely present in the laboratory. Transfection of liver stage-derived P. berghei parasites should enable generation of transgenic parasites within 8-18 days.


Long-term stability of human aflatoxin B1 albumin adducts assessed by isotope dilution mass spectrometry and high-performance liquid chromatography-fluorescence.
The measurement of the aflatoxin B(1)-lysine serum albumin adduct in human blood samples is the most facile biomarker for the assessment of chronic exposure to aflatoxin B(1). Many technologies have been developed for the measurement of this protein adduct including immunoassays, high-performance liquid chromatography (HPLC) with fluorescence detection, and a newly developed isotope-dilution mass spectrometry method. Irrespective of the technology used to determine this adduct level, an important question remains about the long-term stability of this damage product in stored samples. To address this issue, 19 human serum samples that had been previously analyzed for the aflatoxin B(1)-lysine adduct by high-performance liquid chromatography-fluorescence in 1989 were re-analyzed by isotope dilution mass spectrometry after storage at -80 degrees C. The adduct concentrations measured by these two techniques were identical within 4% over the range 5 to 100 pg of aflatoxin B(1)-lysine/mg albumin. In addition, the specific chemical structure of the aflatoxin B(1)-lysine adduct in human samples was confirmed for the first time by collision-induced dissociation full scan mass spectrometry analysis of the protonated adduct molecular ion. These results illustrate that the aflatoxin B(1)-lysine serum albumin adduct can be stable in human serum stored at -80 degrees C since 1989, and this provides confidence for the measurement of this biomarker in repository samples from epidemiologic investigations.


Alterations of the medullary epithelial compartment in the Aire-deficient thymus: implications for programs of thymic epithelial differentiation.
A widely held model of thymic epithelial differentiation is based on patterns of keratin expression, where a K8(+)K5(+) progenitor gives rise to K8(+)K5/K14(-) cortical thymic epithelium (CTEC), and medullary thymic epithelium (MTEC) are K8(-)K5(+)K14(+). The thymic phenotype of p63-deficient mice indicates that p63 is an important regulator of proximal stages of thymic epithelial differentiation. In this study, we have examined several features of the thymic medullary compartment in wild-type and Aire-deficient thymi in an effort to integrate the proapoptotic activity of Aire with these different perspectives of TE differentiation. Patterns of keratin and p63 expression by MTEC described here are difficult to reconcile with postmitotic MTEC that express a K8(-)K14(+) phenotype and suggest that the patterns of p63 and keratin expression reflecting differentiation programs of other epithelial tissues provide a useful framework for revising models of TE differentiation. Alterations of the Aire(-/-) MTEC compartment included reduced expression of p63, increased frequency of MTEC expressing truncated Aire protein, and shifts in the pattern of keratin expression and epithelial morphology. These data suggest a scenario where cellular targets of Aire-mediated apoptosis are postmitotic MTEC that have not yet completed their terminal differentiation program. According to this view, the minor population of globular K8(+)K14(-/low) MTEC observed in the Aire(+/+) thymus and significantly expanded in the Aire(-/-) thymic medulla represent end-stage, terminally differentiated MTEC. These Aire-dependent alterations of the MTEC compartment suggest that the activity of Aire is not neutral with respect to the program of MTEC differentiation.


The influence of MHC gene products on the generation of an antigen-specific T-cell repertoire.
We have previously described the B10.A pigeon cytochrome c-specific response in terms of clonal phenotypes and T-cell receptor (TcR) gene usage. All B10.A T-cell clones studied respond to antigen in association with syngeneic B10.A APCs and cross-react to antigen in association with one or two allogeneic variants of the I-E-encoded MHC molecules. In congenic strains of mice expressing these allogeneic MHC alleles [B10.A(5R) and B10.S(9R)], pigeon cytochrome c-specific T cells exhibit very similar MHC cross-reactivities. Our goal was to determine whether the same MHC cross-reactive T-cell clones were expressed in each appropriate strain, or whether each T-cell repertoire was unique. The results indicate that identical V alpha-J alpha and V beta-J beta combinations were expressed by the major pigeon cytochrome c-specific response phenotype in B10.A and B10.A(5R) mice. Previous functional data supports this overlap in expressed T-cell clones. B10.A and B10.S(9R) mice exhibit similar response phenotypes to pigeon cytochrome c but express distinctly different TcR genes. The results of these studies support the existence of at least two different mechanisms in determining MHC-linked immune response polymorphisms.


Reduction of dietary obesity in aP2-Ucp transgenic mice: mechanism and adipose tissue morphology.
C57BL6/J mice with the expression of the mitochondrial uncoupling protein (UCP) gene from the fat-specific aP2 gene promoter were used to study the mechanism by which the aP2-Ucp transgene affects adiposity and reduces high-fat diet induced obesity. In the transgenic mice, UCP synthesized in white fat was inserted into mitochondria, and oxygen uptake by epididymal fat fragments indicated UCP-induced thermogenesis. The respirometry data, UCP content, cytochrome oxidase activity, and tissue morphology suggested functional involution of brown fat. Despite 25- to 50-fold lower mitochondrial cytochrome oxidase activity in white than in brown fat cells, total oxidative capacity in white and brown adipose tissue is comparable. Appearance of novel small cells in the gonadal fat of the transgenic mice was associated with a higher DNA content than that of the nontransgenic mice. The results prove a potential of transgenically altered mitochondria in white fat to modulate adiposity and energy expenditure and suggest the existence of a yet unidentified site-specific link between energy metabolism in adipocytes and cellularity.


X-ray micro-tomography of Carboniferous stem-Dictyoptera: new insights into early insects.
Computer reconstructions of Archimylacris eggintoni, a Carboniferous stem-group dictyopteran ('roachoid'), are presented. A siderite-hosted specimen was scanned using high-resolution X-ray microtomography (µCT), and a 'virtual fossil' was created with a resolution of 17.7 µm. This has revealed the morphology in great detail, including adhesive limb structures indicative of climbing and specializations for rapid movement. The antennae are filiform, and the mandibles are comparable to those of certain extant cockroaches, suggesting a similar generalist, saprophagous diet. The reconstruction reveals a high degree of specialization, and provides insights into the mode of life of these common Palaeozoic insects. Further µCT study of insect fossils has the potential to supplement wing venation with new characters, and hence improve fossil insect phylogenies.


Pseudoceranoids A-J, Sesquiterpene-Based Meroterpenoids with Cytotoxicity from the Sponge Pseudoceratina purpurea.
Pseudoceranoid A (1), a rare merosesquiterpene featuring a rearranged 4,9-friedodrimane-type core with a crotonolactone moiety, two new rearranged 4,9-friedodrimane-type sesquiterpene cyclopentanones (2 and 3), and three new rearranged 4,9-friedodrimane-type sesquiterpene hydroquinones (4-6), along with two new drimane-type sesquiterpene derivatives (7 and 8), as well as two new 4,9-friedodrimane-type sesquiterpene quinones (9 and 10), were isolated from the South China Sea sponge Pseudoceratina purpurea. The structures of compounds were established by analysis of spectroscopic data, as well as by single-crystal X-ray diffraction, DP4+ probability analyses, and calculated electronic circular dichroism. Compound 4 showed weak cytotoxicity against K562, H69AR, and MDAMB-231 cell lines with IC50 values of 3.01, 7.74, and 9.82 μM, respectively. Compound 5 exhibited cytotoxicity against the H69AR cell line with an IC50 value of 2.85 μM.


Successful use of platelet-rich plasma for chronic plantar fasciitis.
There are limited data on the use of platelet-rich plasma (PRP) for treating chronic plantar fasciitis. The purpose of this study was to document the clinical outcomes of patients who were treated with PRP injections for plantar fasciitis to determine the degree to which injections were able to decrease the visual analogue scale (VAS) pain scores and improve patient reported functional scores. This was a retrospective review of 23 consecutive patients treated with PRP for chronic plantar fasciitis (symptoms lasting over 6 months). Patients returned after 4 weeks for a postinjection follow-up. A second injection was given if significant improvement was not obtained by that time. Postinjection foot and ankle outcome scores (FAOS), 12-item short form health survey (SF-12), and VAS scores were collected at a minimum of 6 months follow-up. Thirty injections were given in 23 patients, with one patient lost to follow-up. The mean VAS score improved from 7 to 4. The pain, symptoms, and quality of life subscales of the FAOS and SF-12 significantly improved from preinjection scores. Five patients went on to have endoscopic release of the plantar fascia at an average of 94 days after the last injection (range, 22-314 days). Six patients obtained full resolution of symptoms while the majority of patients were able to forgo surgery due to improvement from the PRP injection. These results provide preliminary information on the safety and efficiency of PRP injection as treatment for chronic plantar fasciitis.


Treatment of segmental bone defects in rats by the stimulation of bone marrow osteo-progenitor cells with prostaglandin E2.
An alternative to bone grafting is engineered osteo-conductive material that carries osteo-progenitor cells with osteo-stimulant factors impregnated on a malleable osteo-conductive material. We used bone marrow stem cells as the source of osteo-progenitor cells and stimulated them with prostaglandin E2 using demineralised bone matrix as a carrier. We treated 35 skeletally mature male Wistar albino rats with segmentary radial bone defects using five different treatment groups. Group I received no treatment; the remaining four groups all received a mixture of bone marrow and demineralised bone matrix. In group III, a copolymer was added. In group IV, prostaglandin E2 and in group V prostaglandin E2 within a copolymer was added to the mixture. Eight weeks after the surgical procedure, the rats were sacrificed. Radiological and histological evaluation of the radial bone showed that while there was no significant healing in groups I, II and III, there was a significant healing response in groups IV and V.


Percutaneous nephrolithotomy of kidney calculi in horseshoe kidney.
The aim of this study was to evaluate percutaneous nephrolithotomy (PCNL) in horseshoe kidneys with calculi. Between 1995 and 2005, we performed PCNL in 9 patients with horseshoe kidney. In 3 of them, there was a single calculus and the rest had multiple calculi in the pelvis and at least 1 in the calyxes. Ultrasonography, plain abdominal radiography, and intravenous urography (IVU) were performed in all patients. We used fluoroscopy for entering the system and then, pneumatic or ultrasonic lithotripsy was used. In all except 1 patient (88.9%) we could access the system. Single calculi in 3 patients were removed. In 5 patients with multiple calculi, the calculus causing obstruction was removed, and in 3, the calculi located in the calyxes were removed too. Consequently, 66.7% were stone-free at the end of the procedure. In 2 patients, there were residual calculi in the calyxes and they underwent candidates for extracorporeal shockwave lithotripsy. Percutaneous nephrolithotomy can be used in patients with horseshoe kidney if the patient selection is appropriate and the surgeon is experienced enough. The success rate and complications are the same as the patients with normal anatomy. However, access to the lower calyx is more difficult due to its anatomic status.


Amebic perforation of the colon.
Acute necrotising colitis and perforation of the colon due to amebic infection is usually fatal. Only three patients in the present series of nine survived surgery. Preoperative diagnosis was not possible and because of an extremely friable, blotting paper like colon, this condition was only suspected at laparotomy. Examination of ulcer scrapings for the presence of Entamoeba histolytica or histopathology is confirmatory. Resection anastomosis is mandatory as less extensive local procedures or bypass surgery are not always feasible.


Invited Review: Long non-coding RNAs: important regulators in the development, function and disorders of the central nervous system.
Genome-wide transcriptional studies have demonstrated that tens of thousands of long non-coding RNAs (lncRNA) genes are expressed in the central nervous system (CNS) and that they exhibit tissue- and cell-type specificity. Their regulated and dynamic expression and their co-expression with protein-coding gene neighbours have led to the study of the functions of lncRNAs in CNS development and disorders. In this review, we describe the general characteristics, localization and classification of lncRNAs. We also elucidate the examples of the molecular mechanisms of nuclear and cytoplasmic lncRNA actions in the CNS and discuss common experimental approaches used to identify and unveil the functions of lncRNAs. Additionally, we provide examples of lncRNA studies of cell differentiation and CNS disorders including CNS injuries and neurodegenerative diseases. Finally, we review novel lncRNA-based therapies. Overall, this review highlights the important biological roles of lncRNAs in CNS functions and disorders.


Canine cryptorchism and subsequent testicular neoplasia: case-control study with epidemiologic update.
A retrospective study of 2,912 cryptorchid dogs identified 14 breeds with significantly high risk. Among six distinct closely interrelated breed groups (e.g., toy, miniature, and standard poodles), the risk in the smaller breed was always greater than that in the larger relative, suggesting that genetically influenced maldescent could be, in part, related to physical size or the rate of growth of the involved structures. Testicular tumors were diagnosed in 5.7% of the cryptorchid dogs; half had only Sertoli cell tumors, one-third had only seminomas. The relative risk for Sertoli cell tumor or seminoma was not directly related to a familial risk for cryptorchism. Using the health experience of a control population composed of male dogs with anal sac disease (N = 4,184), there is an estimated relative risk of 9.2 in cryptorchid dogs to develop a testis tumor (95% confidence interval, 5.9-14.3) and 4.2 in dogs with inguinal hernia (95% confidence interval, 1.8-9.5). Considering that the anatomical development of the genital tract, testis descent, and tunic relationships in dog are very similar to that in man, and that the associations of cryptorchism and inguinal hernia with testis neoplasms are also similar, the dog should be an excellent model system to further investigate the causes of human cryptorchism.


Oxidative stress in patients with rheumatoid arthritis.
Rheumatoid arthritis is an autoimmune disease of unknown etiology, characterized by articular inflammation. Oxidative damage induced by reactive oxygen species has been related to the pathophysiology of rheumatoid arthritis in several studies, although results have been inconsistent and contradictory. To determine oxidative stress markers in patients with rheumatoid arthritis. Descriptive cross-sectional study in rheumatoid arthritis patients and healthy controls. In peripheral blood samples from all study subjects, lipid peroxide (thiobarbituric acid reactive substances) and protein carbonyl levels were quantified as oxidative damage markers; superoxide dismutase and glutathione peroxidase activities, glutathione concentration, and the reduced glutathione/oxidized glutathione ratio were analyzed as antioxidant defense indicators. Mann-Whitney U tests were run. Statistical significance (a) was 0.05%. We included 29 rheumatoid arthritis patients, 10 with active disease, and 41 healthy controls. Age was higher in the rheumatoid arthritis group; there were no differences in female:male ratio between groups. Oxidative damage was higher in rheumatoid arthritis patients; however, there was no difference between patients with active or inactive rheumatoid arthritis. Antioxidant enzyme activities, glutathione concentration, and reduced glutathione/oxidized glutathione ratio were higher in rheumatoid arthritis patients than in controls. Antioxidant levels were higher in rheumatoid arthritis patients than in healthy controls; however, they were insufficient to prevent oxidative damage. This suggests an active oxidative process in rheumatoid arthritis patients.


Imaging-guided costotransversectomy for thoracic disc herniation.
The surgical management of thoracic disc disease remains challenging. Outcomes after laminectomy had been poor, and modern posterolateral, lateral, and anterior approaches have evolved to replace this older procedure. Each has its own set of complications, and all are hampered, to varying degrees, by the limited visualization of the ventral disc space and spinal cord during decompression. The authors present their early experience with computer-assisted image guidance as an adjunctive tool for preoperative planning and navigation in the treatment of thoracic disc disease. Five consecutive patients underwent image-guided costotransversectomies between January 1999 and April 2000. The levels of herniation were T8-9 in three and T7-8 and T5-6, respectively, in the other two. There were four centrolateral herniations and one midline herniation. Three discs were soft and two hard. Two patients had previously undergone failed disc excisions. All patients had axial pain and myeloradiculopathies preoperatively. Three were unable to walk. Four patients enjoyed good or excellent outcomes, with return of ambulation. One patient experienced only mild improvement in her severe paraparesis. Image-guidance was invaluable in planning the corpectomy and aiding visualization in situations in which the dura or disc were obscured; its use allowed successful surgical excisions in the most challenging circumstances.


Activities of vancomycin, ceftaroline, and mupirocin against Staphylococcus aureus isolates collected in a 2011 national surveillance study in the United States.
Forty-two medical centers from throughout the United States participating in a longitudinal surveillance program were asked to submit 100 consecutive Staphylococcus aureus isolates during July to December 2011. Susceptibility testing using CLSI broth microdilution and mecA detection by PCR analysis was performed on the 4,131 isolates collected. Methods employing Etest glycopeptide resistance detection (GRD; bioMérieux) and brain heart infusion agar containing 4 μg/ml vancomycin (BHIV) were used to screen methicillin-resistant S. aureus (MRSA) isolates for heterogeneous intermediate-level resistance to vancomycin (hVISA). Isolates with positive hVISA screen results were confirmed by population analysis profiling-area under the curve (PAP-AUC) determinations. The genetic relatedness of hVISA, ceftaroline-nonsusceptible, or high-level (HL) mupirocin resistance MRSA isolates was assessed by pulsed-field gel electrophoresis (PFGE). Among 2,093 MRSA isolates, the hVISA screen results were positive with 47 isolates by Etest GRD and 30 isolates by BHIV agar screen. Twenty-five of the GRD- or BHIV screen-positive isolates were confirmed as hVISA by PAP-AUC testing. Results of the current study were compared to results obtained from prior surveillance performed in 2009. The prevalence of hVISA among MRSA isolates was higher in 2011 than in 2009 (1.2% versus 0.4%, P = 0.003), especially for isolates with a vancomycin MIC of 2 (45.4% versus 14.3%, P = 0.01). The overall rate of ceftaroline susceptibility in the current study was 99.4% (one hVISA isolate had an intermediate ceftaroline MIC). HL mupirocin resistance increased from 2.2% in 2009 to 3.2% in 2011 (P = 0.006). Although overall rates of hVISA and HL mupirocin resistance are low, they have increased since 2009.


An integrated method to evaluate and monitor the conservation state of coralligenous habitats: The INDEX-COR approach.
A new method based on photographic sampling coupled with in situ observations was applied to 53 stations along the French Mediterranean coast, to assess the integrity of coralligenous reefs affected by different levels of anthropogenic pressure. The conservation state of the assemblages characterizing these habitats was then assessed by an index - the INDEX-COR - that integrates three metrics: (i) the sensitivity of the taxa to organic matter and sediment deposition, (ii) the observable taxonomic richness, and (iii) the structural complexity of the assemblages. The sensitivity of INDEX-COR was tested and showed good correlation with the Level of Pressure calculated for each station according to expert judgment and field observations.


Left ventricular aneurysmectomy.
Comparison of preoperative and postoperative studies in 81 patients undergoing left ventricular aneurysmectomy failed to show consistent hemodynamic trends. The most reliable prognostic indicator for survival (84% early, 71% late) was the function of the basilar ventricular segments. In 62 of the 81 patients, there was concomitant aortocoronary bypass grafting. Eighty-eight percent of the surviving patients are essentially free of symptoms. These findings support the continued surgical treatment of ventricular aneurysm in symptomatic patients, and suggest nonoperative treatment for patients who are asymptomatic.


High-density diffuse optical tomography of term infant visual cortex in the nursery.
Advancements in antenatal and neonatal medicine over the last few decades have led to significant improvement in the survival rates of sick newborn infants. However, this improvement in survival has not been matched by a reduction in neurodevelopmental morbidities with increasing recognition of the diverse cognitive and behavioral challenges that preterm infants face in childhood. Conventional neuroimaging modalities, such as cranial ultrasound and magnetic resonance imaging, provide an important definition of neuroanatomy with recognition of brain injury. However, they fail to define the functional integrity of the immature brain, particularly during this critical developmental period. Diffuse optical tomography methods have established success in imaging adult brain function; however, few studies exist to demonstrate their feasibility in the neonatal population. We demonstrate the feasibility of using recently developed high-density diffuse optical tomography (HD-DOT) to map functional activation of the visual cortex in healthy term-born infants. The functional images show high contrast-to-noise ratio obtained in seven neonates. These results illustrate the potential for HD-DOT and provide a foundation for investigations of brain function in more vulnerable newborns, such as preterm infants.


N.A.S. symposium: new evidence as the basis for increased efforts in cancer research.
Recent studies, primarily with mouse, rat, and chicken cells, have provided evidence to support the concept that vertebrates contain the genetic information for producing a type-C RNA tumor virus in an unexpressed form in their somatic cells as well as in their germ cells. This information, which our associates and we postulated has been part of the genetic makeup of vertebrates since early in evolution, can persist for hundreds of generations in cell culture without overt production of virus. It is proposed that the endogenous virogenes (the genes for the production of type-C viruses) and the oncogenes (that portion of the virogene responsible for transforming a normal cell into a tumor cell) are maintained in an unexpressed form by repressors in normal cells. Various agents, including radiation, chemical carcinogens, and, perhaps, exogenously added viruses, may transform cells by "switching on" the endogenous oncogenic information. Some other implications of the viral oncogene theory are presented.


Effects of long-term treatment with Dienogest on the quality of life and sexual function of women affected by endometriosis-associated pelvic pain.
The aim of this prospective study was to evaluate quality of life (QoL) and sexual function of women affected by endometriosis pain treated with Dienogest (DNG) for 24 months. Fifty-four women constituted the study group and were given DNG 2 mg/daily; 38 women were given non-steroidal anti-inflammatory drugs (NSAIDs) and constituted the control group. To define endometriosis-associated pelvic pain, dysmenorrhea and dyspareunia the Visual Analogic Scale (VAS) was used. The Short Form-36 (SF-36), the Female Sexual Function Index (FSFI) and the Female Sexual Distress Scale (FSDS) were used to assess the QoL, sexual function and the sexual distress, respectively. The study included five follow-ups at 3, 6, 12, 18 and 24 months. Slight improvements in chronic pelvic pain, dysmenorrhea and dyspareunia were observed in the study group at 3 months (p<0.05) and improved more from 6 to 24 months of DNG treatment (p<0.001). QoL improved with a similar trend: at 3 months the improvement was significant in several categories (p<0.05), and from 6 to 24 months in all categories (p<0.001). The FSFI score did not change at the 3 month follow-up (p=not significant [NS]) but it improved from 6 to 24 months (p<0.001). A similar trend was observed for the FSDS score (p<0.001). No change was observed in the control group (p=NS). Long-term treatment with DNG 2 mg once-daily in women with endometriosis-associated pelvic pain may have positive effects on the QoL and sexual life, confirming the observations of the previous study on the first 6 months of treatment.


Queries to the British Orthodontic Society.
As the representative body for orthodontic specialists, the British Orthodontic Society (BOS) provides advice and guidance to professionals and patients on a variety of topics. The BOS receive frequent queries from professionals and patients and, where appropriate, provide advice and guidance. By reflecting on the types of queries received, it is hoped that general guidance can be developed and 'frequently asked questions' information established to improve knowledge in these particular areas. Retrospective assessment of queries raised to the BOS. BOS headquarters, London. All queries raised to the BOS by professionals and patients from January 2017 to October 2018 were collated and divided into those raised by dental professionals or by patients. Within each group, all queries were further subcategorised into different topic areas. For each of the main topics, guidance will be developed to improve clinician and patient knowledge in these areas. In the time period assessed, 422 queries were received to the BOS, 51.8% (n = 229) were from patients and 48.2% (n = 213) from professionals. Commissioning was the most common topic queried by professionals (34.3%), which included NHS contracts and tendering. Queries regarding associate matters (9.4%), specific clinical queries (9.4%) and BOS issues (8.9%) were also common topics raised by professionals. The most common topic queried by patients was the eligibility for NHS treatment (29.3%), followed by specific clinical queries (22.3%) and seeking recommendations for clinicians (10.5%). Overall, a large number of queries were received by the BOS. While some queries required specific information from the BOS, the majority of queries could be addressed through already published information and guidance. This project identifies the more common queries and provides clear guidance on where to find the appropriate advice.


Phosphorylation of Not4p functions parallel to BUR2 to regulate resistance to cellular stresses in Saccharomyces cerevisiae.
The evolutionarily conserved Ccr4-Not and Bur1/2 kinase complexes are functionally related in Saccharomyces cerevisiae. In this study, we further explore the relationship between the subunits Not4p and Bur2p. First, we investigated the presence of post-translational modifications on the Ccr4-Not complex. Using mass spectrometry analyses we identified several SP/TP phosphorylation sites on its Not4p, Not1p and Caf1p subunits. Secondly, the influence of Not4p phosphorylation on global H3K4 tri-methylation status was examined by immunoblotting. This histone mark is severely diminished in the absence of Not4p or of Bur2p, but did not require the five identified Not4p phosphorylation sites. Thirdly, we found that Not4p phosphorylation is not affected by the kinase-defective bur1-23 mutant. Finally, phenotypic analyses of the Not4p phosphomutant (not4S/T5A) and bur2Delta strains showed overlapping sensitivities to drugs that abolish cellular stress responses. The double-mutant not4S/T5A and bur2Delta strain even revealed enhanced phenotypes, indicating that phosphorylation of Not4p and BUR2 are active in parallel pathways for drug tolerance. Not4p is a phospho-protein with five identified phosphorylation sites that are likely targets of a cyclin-dependent kinase(s) other than the Bur1/2p complex. Not4p phosphorylation on the five Not4 S/T sites is not required for global H3K4 tri-methylation. In contrast, Not4p phosphorylation is involved in tolerance to cellular stresses and acts in pathways parallel to BUR2 to affect stress responses in Saccharomyces cerevisiae.


Update on the optimal management of patients with colorectal liver metastases.
Patients with colorectal liver metastases represent a distinct subset of metastatic colorectal cancer. Optimal management requires a multidisciplinary approach, local and systemic. Curative hepatic surgery is standard for resectable cases, but unfortunately, the majority of patients are not initially resectable due to the size, location, and/or extent of disease, inadequate remnant liver volume, or comorbidities. Other local approaches may be complementary (such as portal vein embolization) or alternative (such as ablation, hepatic arterial infusion, selective internal radiation therapy, and stereotactic body radiotherapy) to surgery. Systemic therapy can downsize disease, allowing surgical resection and, potentially, long-term survival, but it must be balanced against the potential for hepatotoxicity. Current standard approaches including cytotoxics and biologics, such as bevacizumab and particularly anti-epidermal growth factor receptor therapy, improve response rates and may enhance downsizing and resection rates. Optimization of local therapies and systemic conversion strategies via controlled, randomized trials is still a pending question.


Progression from paroxysmal to persistent atrial fibrillation clinical correlates and prognosis.
We investigated clinical correlates of atrial fibrillation (AF) progression and evaluated the prognosis of patients demonstrating AF progression in a large population. Progression of paroxysmal AF to more sustained forms is frequently seen. However, not all patients will progress to persistent AF. We included 1,219 patients with paroxysmal AF who participated in the Euro Heart Survey on AF and had a known rhythm status at follow-up. Patients who experienced AF progression after 1 year of follow-up were identified. Progression of AF occurred in 178 (15%) patients. Multivariate analysis showed that heart failure, age, previous transient ischemic attack or stroke, chronic obstructive pulmonary disease, and hypertension were the only independent predictors of AF progression. Using the regression coefficient as a benchmark, we calculated the HATCH score. Nearly 50% of the patients with a HATCH score >5 progressed to persistent AF compared with only 6% of the patients with a HATCH score of 0. During follow-up, patients with AF progression were more often admitted to the hospital and had more major adverse cardiovascular events. A substantial number of patients progress to sustained AF within 1 year. The clinical outcome of these patients regarding hospital admissions and major adverse cardiovascular events was worse compared with patients demonstrating no AF progression. Factors known to cause atrial structural remodeling (age and underlying heart disease) were independent predictors of AF progression. The HATCH score may help to identify patients who are likely to progress to sustained forms of AF in the near future.


Structure, organization, and transcription units of the human alpha-platelet-derived growth factor receptor gene, PDGFRA.
Isolation and characterization of genomic clones encoding human alpha-platelet derived growth factor receptor (HGMW-approved symbol PDGFRA) revealed that the gene spans approximately 65 kb and contains 23 exons. The 5'-untranslated region of the mRNA is encoded by exon 1, and a large intron of 23 kb separates exon 2 encoding the translation initiator codon AUG and the signal sequence. The locations of exon/intron boundaries in the extracellular immunoglobulin-like domains, the transmembrane domain, the two cytoplasmic tyrosine kinase domains, and the kinase insertion domain are very similar to those in c-kit and macrophage colony stimulating factor-1 receptor genes. The transcription start site was mapped to a position 393 bp upstream of the AUG translocation initiator codon by S1 mapping and primer extension analysis. The 5'-flanking region of the gene lacks a typical TATA box but contains a typical CCAAT box and GATA motifs. This region also contains potential sites for AP-1, AP-2, Oct-1, Oct-2, and Sp1. The 5'-flanking region of the gene was fused to the luciferase reporter gene, and transcription units of the gene were determined.


Modified prone position to dissect the popliteal fossa.
Traditionally, the prone position is used for dissecting the popliteal fossa, which requires endotracheal intubation. Access to the airway in this position is limited, hence the complications. It is not surprising that the prone position is not favoured by the anaesthetists, especially in patients with a high body mass index. We describe a safe and novel alternative to the prone position. The modified prone position (MPP) is described as an alternative position that facilitates access to the airway. Between October 2007 and May 2010, 12 patients underwent popliteal fossa dissection using the MPP. All patients had general anaesthesia using a laryngeal mask airway with the exception of one, who had an epidural anaesthesia. There were no airway or haemodynamic complications. The surgical access to the popliteal fossa was as good as with the traditional prone position. The MPP was satisfactory for both the surgeon and the anaesthetists. The authors now use this position routinely for dissecting the popliteal fossa.


Cryopreservation in trehalose preserves functional capacity of murine spermatogonial stem cells.
Development of techniques to isolate, culture, and transplant human spermatogonial stem cells (SSCs) has the future potential to treat male infertility. To maximize the efficiency of these techniques, methods for SSC cryopreservation need to be developed to bank SSCs for extended periods of time. Although, it has been demonstrated that SSCs can reinitiate spermatogenesis after freezing, optimal cryopreservation protocols that maximize SSC proliferative capacity post-thaw have not been identified. The objective of this study was to develop an efficient cryopreservation technique for preservation of SSCs. To identify efficient cryopreservation methods for long-term preservation of SSCs, isolated testis cells enriched for SSCs were placed in medium containing dimethyl sulfoxide (DMSO) or DMSO and trehalose (50 mM, 100 mM, or 200 mM), and frozen in liquid nitrogen for 1 week, 1 month, or 3 months. Freezing in 50 mM trehalose resulted in significantly higher cell viability compared to DMSO at all thawing times and a higher proliferation rate compared to DMSO for the 1 week freezing period. Freezing in 200 mM trehalose did not result in increased cell viability; however, proliferation activity was significantly higher and percentage of apoptotic cells was significantly lower compared to DMSO after freezing for 1 and 3 months. To confirm the functionality of SSCs frozen in 200 mM trehalose, SSC transplantation was performed. Donor SSCs formed spermatogenic colonies and sperm capable of generating normal progeny. Collectively, these results indicate that freezing in DMSO with 200 mM trehalose serves as an efficient method for the cryopreservation of SSCs.


Making a friend from a foe: expressing a GroEL gene from the whitefly Bemisia tabaci in the phloem of tomato plants confers resistance to tomato yellow leaf curl virus.
Some (perhaps all) plant viruses transmitted in a circulative manner by their insect vectors avoid destruction in the haemolymph by interacting with GroEL homologues, ensuring transmission. We have previously shown that the phloem-limited begomovirus tomato yellow leaf curl virus (TYLCV) interacts in vivo and in vitro with GroEL produced by the whitefly vector Bemisia tabaci. In this study, we have exploited this phenomenon to generate transgenic tomato plants expressing the whitefly GroEL in their phloem. We postulated that following inoculation, TYLCV particles will be trapped by GroEL in the plant phloem, thereby inhibiting virus replication and movement, thereby rendering the plants resistant. A whitefly GroEL gene was cloned in an Agrobacterium vector under the control of an Arabidopsis phloem-specific promoter, which was used to transform two tomato genotypes. During three consecutive generations, plants expressing GroEL exhibited mild or no disease symptoms upon whitefly-mediated inoculation of TYLCV. In vitro assays indicated that the sap of resistant plants contained GroEL-TYLCV complexes. Infected resistant plants served as virus source for whitefly-mediated transmission as effectively as infected non-transgenic tomato. Non-transgenic susceptible tomato plants grafted on resistant GroEL-transgenic scions remained susceptible, although GroEL translocated into the grafted plant and GroEL-TYLCV complexes were detected in the grafted tissues.


Induction and modulation of lung tumors: genomic and transcriptional alterations in cigarette smoke-exposed mice.
Cigarette smoke plays a major role in the epidemiology of lung cancer, and smoke components have extensively been investigated in carcinogenicity and chemoprevention studies in experimental animals. However, it is much more difficult to reproduce the tumorigenicity of the whole complex mixture in preclinical models. The authors review here some results obtained in their laboratories, dealing with the induction of lung tumors, and genomic and transciptional alterations in smoke-exposed mice. The authors were successful in inducing lung tumors in 4 strains of mice exposed whole-body to environmental cigarette smoke, including Swiss albino, A/J, SKH-1 hairless, and p53 mutant (UL533 x A/J)F1 mice. However, the tumorigenic response was rather weak in all strains. Much more intense were the smoke-induced alterations of a variety of intermediate biomarkers, such as cytogenetic end points in pulmonary alveolar macrophages, bone marrow and peripheral blood erythrocytes; apoptosis, p53 oncoprotein, and proliferating cell nuclear antigen in the bronchial epithelium; bulky DNA adducts, 8-hydroxy-2-deoxyguanosine; multigene expression, and thiobarbituric acid-reactive aldehydes in whole lung and several other organs. Smoke-induced genomic and transcriptional alterations were suitable for evaluating their modulation by chemopreventive agent, as shown in studies using the thiol N-acetylcysteine and the nonsteroidal anti-inflammatory drug sulindac.


I didn't feel like drinking but I don't know why: the effects of evaluative conditioning on alcohol-related attitudes, craving and behavior.
The goal of the present research was to test the value of evaluative conditioning (EC) to unobtrusively change alcohol-related attitudes and drinking behavior. In the EC paradigm, participants had to spot an irrelevant target picture in a series of trials in which many different stimuli were presented. In the experimental condition, beer-related pictures (CSs) were consistently paired with negative words and pictures (USs) in a number of trials. In the control condition, participants were exposed to the same stimuli, but without the critical alcohol-negative pairings. After the EC task, participants participated in an allegedly second experiment in which we measured beer-related attitudes, craving for beer, and actual drinking behavior both during a bogus taste test and during the week following the experiment. Compared to participants in the control condition, participants in the experimental condition showed more negative attitudes toward beer, experienced less craving for beer, and consumed less beer both in the lab during the taste test and outside the lab during the week following the manipulation. These findings suggest that unhealthy drinking behavior may be targeted through EC procedures.


Aspergillus penicillioides Speg. Implicated in Keratomycosis.
The aim of the study was mycological examination of ulcerated corneal tissues from an ophthalmic patient. Tissue fragments were analyzed on potato-glucose agar (PDA) and maltose (MA) (Difco) media using standard laboratory techniques. Cultures were identified using classical and molecular methods. Macro- and microscopic colony morphology was characteristic of fungi from the genus Aspergillus (restricted growth series), most probably Aspergillus penicillioides Speg. Molecular analysis of the following rDNA regions: ITS1, ITS2, 5.8S, 28S rDNA, LSU and β-tubulin were carried out for the isolates studied. A high level of similarity was found between sequences from certain rDNA regions, i.e. ITS1-5.8S-ITS2 and LSU, what confirmed the classification of the isolates to the species A. penicillioides. The classification of our isolates to A. penicillioides species was confirmed also by the phylogenetic analysis. The aim of the study was mycological examination of ulcerated corneal tissues from an ophthalmic patient. Tissue fragments were analyzed on potato-glucose agar (PDA) and maltose (MA) (Difco) media using standard laboratory techniques. Cultures were identified using classical and molecular methods. Macro- and microscopic colony morphology was characteristic of fungi from the genus Aspergillus (restricted growth series), most probably Aspergillus penicillioides Speg. Molecular analysis of the following rDNA regions: ITS1, ITS2, 5.8S, 28S rDNA, LSU and β-tubulin were carried out for the isolates studied. A high level of similarity was found between sequences from certain rDNA regions, i.e. ITS1-5.8S-ITS2 and LSU, what confirmed the classification of the isolates to the species A. penicillioides. The classification of our isolates to A. penicillioides species was confirmed also by the phylogenetic analysis.


Urological complications after simultaneous renal and pancreatic transplantation.
To report the urological complications after simultaneous renal and pancreatic transplantation. Retrospective study. Teaching hospital, Italy. 143 consecutive patients having simultaneous renal and pancreatic transplantation by one of three techniques. 33 segmental pancreas with duct occlusion, 77 whole pancreas with bladder diversion, and 33 enteric diversion with systemic (n = 26) or portal venous drainage (n = 7). Urological complications were related to the pancreatic transplant, to the renal transplant, or unrelated to the transplant. Morbidity. After occlusion of the duct and enteric diversion, there were no urological complications related to the pancreatic transplant. On the other hand, among the 77 patients with pancreatic drainage into the bladder, urological complications were common (56/77; 73%). Complications related to the renal transplant were recorded in 6/33 (18%), 26/77 (34%) and 12/33 (36%), respectively. Complications unrelated to the transplant occurred in 6/77 patients (8%) in the bladder drainage group. Five patients after bladder drainage required cystoenteric conversion. Enteric diversion is a safe alternative to bladder diversion and results in significantly fewer urological complications.


Paratyphoid fever: a report of 62 cases with several unusual findings and a review of the literature.
Previous descriptions of paratyphoid fever (PTF) are of limited value to the clinician who has had no experience with Salmonella enteric fevers. Sixty-two cases of PTF occurred in a healthy, homogeneous population. Findings supported by the literature included a seasonal incidence, initially seen symptoms of gastroenteritis and spiking fevers, and a high percentage of positive blood cultures. Unusual findings included the rarity of rose spots, patients with fever as their only symptom, two patients with transient papilledema, and two others with peripheral blood smears suggestive of acute leukemia. The findings of this study are compared with previous reports. Likely causes for divergent and contradictory information are inclusion of data from non-PTF states and study of populations with altered states of health.


A Pilot Trial of Pioglitazone HCl and Tretinoin in ALS: Cerebrospinal Fluid Biomarkers to Monitor Drug Efficacy and Predict Rate of Disease Progression.
Objectives. To determine if therapy with pioglitazone HCl and tretinoin could slow disease progression in patients with ALS. Levels of tau and pNFH in the cerebrospinal fluid were measured to see if they could serve as prognostic indicators. Methods. 27 subjects on stable doses of riluzole were enrolled. Subjects were randomized to receive pioglitazone 30 mg/d and tretinoin 10 mg/BID for six months or two matching placebos. ALSFRS-R scores were followed monthly. At baseline and at the final visit, lumbar punctures (LPs) were performed to measure cerebrospinal fluid (CSF) biomarker levels. Results. Subjects treated with tretinoin, pioglitazone, and riluzole had an average rate of decline on the ALSFRS-R scale of -1.02 points per month; subjects treated with placebo and riluzole had a rate of decline of -.86 (P = .18). Over six months of therapy, CSF tau levels decreased in subjects randomized to active treatment and increased in subjects on placebo. Further higher levels of pNF-H at baseline correlated with a faster rate of progression. Conclusion. ALS patients who were treated with tretinoin and pioglitazone demonstrated no slowing on their disease progression. Interestingly, the rate of disease progression was strongly correlated with levels of pNFH in the CSF at baseline.


Influence of Perivascular Adipose Tissue on Microcirculation: A Link Between Hypertension and Obesity.
Alterations in microcirculation play a crucial role in the pathogenesis of cardiovascular and metabolic disorders such as obesity and hypertension. The small resistance arteries of these patients show a typical remodeling, as indicated by an increase of media or total wall thickness to lumen diameter ratio that impairs organ flow reserve. The majority of blood vessels are surrounded by a fat depot which is termed perivascular adipose tissue (PVAT). In recent years, data from several studies have indicated that PVAT is an endocrine organ that can produce a variety of adipokines and cytokines, which may participate in the regulation of vascular tone, and the secretory profile varies with adipocyte phenotype and disease status. The PVAT of lean humans largely secretes the vasodilator adiponectin, which will act in a paracrine fashion to reduce peripheral resistance and improve nutrient uptake into tissues, thereby protecting against the development of hypertension and diabetes. In obesity, PVAT becomes enlarged and inflamed, and the bioavailability of adiponectin is reduced. The inevitable consequence is a rise in peripheral resistance with higher blood pressure. The interrelationship between obesity and hypertension could be explained, at least in part, by a cross-talk between microcirculation and PVAT. In this article, we propose an integrated pathophysiological approach of this relationship, in order to better clarify its role in obesity and hypertension, as the basis for effective and specific prevention and treatment.


Outbreak of Aeromonas hydrophila wound infections associated with mud football.
On 16 February 2002, a total of 26 people presented to the emergency department of the local hospital in the rural town of Collie in southwest Western Australia with many infected scratches and pustules distributed over their bodies. All of the patients had participated in a "mud football" competition the previous day, in which there had been ~100 participants. One patient required removal of an infected thumbnail, and another required surgical debridement of an infected toe. Aeromonas hydrophila was isolated from all 3 patients from whom swab specimens were obtained. To prepare the mud football fields, a paddock was irrigated with water that was pumped from an adjacent river during the 1-month period before the competition. A. hydrophila was subsequently isolated from a water sample obtained from the river. This is the first published report of an outbreak of A. hydrophila wound infections associated with exposure to mud.


Evaluating the Content Validity of Four Performance Outcome Measures in Patients with Elective Hip Replacements and Hip Fractures.
To assess the content validity of performance outcome (PerfO) measures for use with patients undergoing hip fracture (HF) surgery and elective total hip replacement (eTHR). This study was a substudy of a broader evaluation of measurement properties of PerfO measures. The PerfO measures assessed were timed up and go (TUG), four-step stair climb (4SC), long stair climb (LSC), and repeated chair stand (RCS). For this substudy, HF and eTHR participants were interviewed to evaluate the relevance and difficulty of each PerfO measure. Qualitative analysis was conducted on interview transcripts, and summaries of coded data were produced to assess saturation. All 18 HF participants related the PerfO measures (TUG, 4SC, and RSC) to activities they completed in daily life, with slight variations in some specific aspects. For the eight eTHR participants, the correspondence between the PerfO measures (TUG, 4SC, and LSC) and activities in daily life varied: all participants saw similarity in the movements for the TUG; most undertook short stair climbs in daily life, but most did not regularly undertake LSC in daily life. Nevertheless, all HF and eTHR participants reported that the PerfO measures were relevant and had a level of difficulty similar to daily activities. This study contributes novel methods that adapt US regulatory guidance for patient-reported outcome measures to the evaluation of PerfO measures. A structured approach was used to explore specific details of each measure and correspondence to everyday life. This study demonstrates how content validity of PerfO measures can be meaningfully assessed.


Intravascular magnetic resonance/radiofrequency may enhance gene therapy for prevention of in-stent neointimal hyperplasia.
We evaluated the potential of using intravascular magnetic resonance (MR)/radiofrequency (RF) to enhance vascular endothelial growth factor (VEGF) gene therapy of in-stent neointimal hyperplasia. By using a catheter-based approach, VEGF/lentivirus was locally transferred into 10 (five paired) bilateral femoral-iliac arteries of five hypercholesterolemic pigs, whereas the right arteries were heated up to approximately 41 degrees C by using an intravascular MR/RF system. Then, identical stents were placed immediately into the bilateral VEGF-targeted arteries to create in-stent neointimal hyperplasia. At day 60 after gene/stent interventions, the targeted arteries were harvested for histological correlation. X-Ray angiography-detectable in-stent stenoses were found in three of the arteries treated with VEGF genes only, whereas there were no in-stent stenoses in arteries treated by using MR/RF-heated VEGF genes. Correlative histological examination confirmed a 138% reduction in average thickness of neointimal hyperplasia in VEGF/RF-treated arteries compared with VEGF-only-treated arteries (P < .01). We report a potential method of using an intravascular MR/RF heating technique to enhance gene therapy of in-stent restenosis.


Biomechanical relationships between the corneal endothelium and Descemet's membrane.
The posterior face of the cornea consists of the corneal endothelium, a monolayer of cuboidal cells that secrete and attach to Descemet's membrane, an exaggerated basement membrane. Dysfunction of the endothelium compromises the barrier and pump functions of this layer that maintain corneal deturgesence. A large number of corneal endothelial dystrophies feature irregularities in Descemet's membrane, suggesting that cells create and respond to the biophysical signals offered by their underlying matrix. This review provides an overview of the bidirectional relationship between Descemet's membrane and the corneal endothelium. Several experimental methods have characterized a richly topographic and compliant biophysical microenvironment presented by the posterior surface of Descemet's membrane, as well as the ultrastructure and composition of the membrane as it builds during a lifetime. We highlight the signaling pathways involved in the mechanotransduction of biophysical cues that influence cell behavior. We present the specific example of Fuchs' corneal endothelial dystrophy as a condition in which a dysregulated Descemet's membrane may influence the progression of disease. Finally, we discuss some disease models and regenerative strategies that may facilitate improved treatments for corneal dystrophies.


Clinical usefulness of urinary CrossLaps as a sensitive marker of bone metabolism.
CrossLaps peptide [Glu-Lys-Ala-His-Asp-Gly-Gly-Arg], a part of the C-telopeptide of the alpha 1-chain of type I collagen of bone, is a recently developed biochemical marker of bone turnover. In this study, the clinical utility of measurement of urinary CrossLaps was investigated in eleven premenopausal women who received a gonadotropin-releasing hormone (GnRH) agonist for 6 months for treatment of adenomyosis (n = 1) or leiomyomas (n = 10). Along with urinary CrossLaps, the levels of various biochemical markers, and serum estradiol, calcitonin and intact parathyroid hormone (i-PTH) were measured, and lumbar spine bone mineral density (BMD) was also monitored before, during, and at the end of the course of GnRH agonist therapy. Apart from CrossLaps, markers of bone resorption tested were urinary pyridinoline, deoxypyridinoline and hydroxyproline. Markers of bone formation tested were serum osteocalcin and bone-specific alkaline phosphatase (B-ALP). Serum estradiol levels decreased to undetectable levels at 2 months of GnRH agonist therapy. The values for all biochemical markers increased significantly throughout the therapy. The degree of an increase in CrossLaps levels was greater than that in all other markers. Mean lumbar spine (L2-L4) BMD was decreased by 7.2% at 6 months of treatment. The percent change in BMD at 6 months of treatment correlated inversely with the percent change in CrossLaps levels from the baseline to 1, 2, and 5 months of treatment. These results indicate that measurement of urinary CrossLaps might be a useful tool to predict the risk of bone loss caused by hypoestrogenism including GnRH agonist therapy.


How painful are shockwave lithotripsy and endoscopic procedures performed at outpatient urology clinics?
Our aim was to investigate the subjective pain felt by patients during shockwave lithotripsy (SWL) and endoscopic procedures such as cystoscopy, retrograde ureteral stenting, retrograde pyelography (RGP), and ureteroscopic lithotripsy performed in an outpatient clinic, and to identify how severe pain during such procedures is. We estimated subjective pain in 984 patients after SWL (186), cystoscopy (489), retrograde ureteral stenting (127), RGP (97), and ureteroscopic lithotripsy (85) performed by a single expert in an outpatient clinic using a prospective questionnaire with a ten point visual analog scale between January 2001 and December 2003. There was no premedication in any procedure except ureteroscopic lithotripsy for which an intramuscular injection of analgesics (pethidine HCl 50 mg) was used. The pain scale score in SWL was 6.62+/-2.27, the highest among the procedures (P<0.05). Pain scores for endoscopies were 4.48+/-2.07 in retrograde ureteral stenting, 3.81+/-2.06 in ureteroscopic lithotripsy, 3.72+/-1.75 in RGP, and 3.08+/-1.95 in cystoscopy. In this study, we observed that patients feel most pain in SWL without anesthesia, and that pain during ureteroscopic lithotripsy under local anesthesia is not high, compared with other endoscopic procedures.


A Histone Deacetylase Inhibitor Suppresses Epithelial-Mesenchymal Transition and Attenuates Chemoresistance in Biliary Tract Cancer.
Epithelial-mesenchymal transition (EMT) is involved in the characteristics of malignancy, such as invasion, metastasis, and chemoresistance. In biliary tract cancer (BTC), EMT is induced by transforming growth factor-beta 1 (TGF-β1). The EMT is reversible; therefore, it is conceivable that it could be related to some epigenetic changes. We focused on histone deacetylase (HDAC) inhibitors as regulators of TGF-β1 signaling, and investigated their effect on EMT and chemoresistance. We employed four BTC cell lines (MzChA-1, gemcitabine-resistant MzChA-1, TFK-1, and gemcitabine-resistant TFK-1) and used vorinostat as the HDAC inhibitor. The relative mRNA expression of an epithelial marker (CDH1) and mesenchymal markers (CDH2, vimentin, SNAI1) were measured by qRT-PCR to evaluate factors associated with EMT. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed to evaluate the chemoresistance of each cell line. In addition, NOD/SCID mice were used to evaluate the effect of vorinostat in vivo. In the parent MzChA-1 and TFK-1 cell lines, TGF-β1 induced EMT and chemoresistance; while vorinostat inhibited the EMT and chemoresistance induced by TGF-β1. In gemcitabine-resistant cell lines that highly expressed TGF-β1, vorinostat inhibited EMT and attenuated chemoresistance. We showed that vorinostat inhibits nuclear translocation of SMAD4 which is a signaling factor of TGF-β1, and this is one of the mechanisms by which vorinostat regulates EMT. We also showed that vorinostat attenuates the binding affinity of SMAD4 to the CDH1-related transcription factors SNAI1, SNAI2, ZEB1, ZEB2, and TWIST. Furthermore, combination therapy with vorinostat and gemcitabine improved survival time in the mice xenografted with gemcitabine resistant MzChA-1 cells. In conclusion, vorinostat regulated TGF-β1-induced EMT and chemoresistance through inhibition of SMAD4 nuclear translocation.


New treatment for urethral strictures.
A new operative technique using synthetic, absorbable mesh for grafting a urethral defect was applied in 7 mongrel male dogs. The ventral half of the urethral circumference with its surrounding corpus spongiosum was excised for a length of 3 to 4 cm. A Dexon mesh of the same dimensions, woven in our laboratory from polyglycolic acid fibers, was sutured to the defective area. A perineal urethrotomy was established, and no splints were left behind. Dogs were studied between two and six months. Retrograde urethrography showed that the operative area healed without strictures or irregularities. Intravenous urography showed no back pressure effects, and cultured urine was always sterile. Histologic examination two months after surgery showed that the urothelium was completely healed, without inflammatory changes or disruption. Suburothelial tissues were replaced by dense collagenous connective tissue. The excised corpus spongiosum did not regenerate. After six months, the area of dense collagen described was diminished in size so that the operative area could be hardly identified except by the absence of corpus spongiosum.


Systematic development of an evidence-based website on preconception care.
In February 2015, the Flemish Minister of Welfare, Public Health and Family launched a website on preconception care: 'gezondzwangerworden.be'. The website was developed in response to the lack of comprehensive communication on preconception care and the inadequate intake of folic acid among Flemish women. Despite the international recommendation to take 400 μg folic acid on a daily basis one month before conception until 12 weeks of pregnancy, studies show a lack of compliance in women wanting to become pregnant. A compilation of evidence was made through reviewing well-established guidelines on preconception and prenatal care. The quality of guidelines was assessed by means of AGREE II. The topics included in the website were selected by an internal committee of 5 experts and an external committee of 16 experts. Content validation was carried out by 40 experts in preconception care or related topics. The above-described procedure resulted in an evidence-based website with a selection of relevant, validated information for both women and men who plan a pregnancy and professionals who are consulted by these people. Evaluation and recommendation: The website is currently attracting a constant number of 100 to 200 visitors a day. The information on folic acid is among the most requested, which is an important finding with regard to the policy objectives on preconception care. More research is needed in order to evaluate the use and effect of the website more thoroughly.


Cardiac Rehabilitation for Women: A Systematic Review of Barriers and Solutions.
Cardiac rehabilitation (CR) services improve various clinical outcomes in patients with cardiovascular disease, but such services are underutilized, particularly in women. The aim of this study was to identify evidence-based barriers and solutions for CR participation in women. A literature search was carried out using PubMed, EMBASE, Cochrane, OVID/Medline, and CINAHL to identify studies that have assessed barriers and/or solutions to CR participation. Titles and abstracts were screened, and then the full-text of articles that met study criteria were reviewed. We identified 24 studies that studied barriers to CR participation in women and 31 studies that assessed the impact of various interventions to improve CR referral, enrollment, and/or completion of CR in women. Patient-level barriers included lower education level, multiple comorbid conditions, non-English native language, lack of social support, and high burden of family responsibilities. We found support for the use of automatic referral and assisted enrollment to improve CR participation. A small number of studies suggest that incentive-based strategies, as well as home-based programs, may contribute to improving CR attendance and completion rates. A systematic approach to CR referral, including automatic CR referral, may help overcome barriers to CR referral in women and should be implemented in clinical practice. However, more studies are needed to help identify the best methods to improve CR attendance and completion of CR rates in women.


The posterior operation in treatment of cervical spondylosis with myelopathy: a long-term follow-up study.
An analytical study of 59 patients who underwent the posterior operation for cervical spondylosis with myelopathy at the London Hospital between 1945 and 1960 has been made. The postoperative period of observation ranged from five to 20 years with an average of 10 years. Initially 36 patients were improved by the operation, 24 considerably so, and in 15 patients further deterioration was prevented. Five years after operation 33 patients were still improved and in five patients co-existent disease had become apparent. Ten years after operation 30 patients had maintained their initial improvement; progression of the disease accounted for a drop of 10% in the success rate initially achieved. No patient in this series developed postoperative instability of the cervical spine. Patients with less than two years' duration of symptoms and with less neurological disability fared better. The importance of a long follow-up period for the proper assessment of results is thus emphasized. It is suggested that the posterior operation has stood the test of time and will continue to maintain a useful place in the treatment of this condition. Some points are outlined for the successful practice of this operation.


Selective reduction of intracellular guanosine 5'-triphosphate pool by 4-carbamoylimidazolium 5-olate in murine tumor cells.
Ehrlich carcinoma and P388 leukemia cells were rendered resistant to 4-carbamoylimidazolium 5-olate (SM-108), and assessments were made of biochemical and pharmacological determinants for the sensitivity to SM-108 using both sensitive and resistant sublines. We observed that the treatment of cells with SM-108 in vitro caused a remarkable decrease in the intracellular guanosine 5'-triphosphate pool level in sensitive but not in resistant sublines. There was no difference in the ability to take up SM-108 between sensitive and resistant sublines, but the cellular conversion of SM-108 to its nucleotide, which is the putative active anabolite of SM-108, proceeded only in sensitive sublines. Enzymological studies revealed that the activity of adenine phosphoribosyltransferase (EC 18.104.22.168), which is believed to conjugate SM-108 with 5-phospho-alpha-D-ribose 1-diphosphate, was very low in the resistant sublines. These results strongly support our previous hypothesis that SM-108 is activated by adenine phosphoribosyltransferase to SM-108-nucleotide which then inhibits hypoxanthine-5'-monophosphate dehydrogenase (EC 188.8.131.52), a key enzyme for the de novo synthesis of guanosine 5'-monophosphate.


Effects of soybean small peptides on rumen fermentation and on intestinal and total tract digestion of luxi yellow cattle.
Four Luxi beef cattle (400±10 kg) fitted with ruminal, duodenal and ileal cannulas were used in a 4×4 Latin square to assess the effects of soybean small peptide (SSP) infusion on rumen fermentation, diet digestion and flow of nutrient in the gastrointestinal tract. The ruminal infusion of SSP was 0 (control), 100, 200 and 300 g/d. Ruminal SSP infusion linearly (p<0.01) and quadratically (p<0.01) increased microbial protein synthesis and rumen ammonia-N concentration. Concentrations of total volatile fatty acid were linearly increased (p = 0.029) by infusion SSP. Rumen samples were obtained for analysis of microbial ecology by real-time PCR. Populations of rumen Butyrivibrio fibrisolvens, Streptococcus bovis, Ciliate protozoa, Ruminococcus flavefaciens, and Prevotella ruminicola were expressed as a proportion of total Rumen bacterial 16S ribosomal deoxyribonucleic acid (rDNA). Butyrivibrio fibrisolvens populations which related to total bacterial 16S rDNA were increased (p<0.05), while Streptococcus bovis populations were linearly (p = 0.049) and quadratically (p = 0.020) decreased by infusion of SSP. Apparent rumen digestibility of DM and NDF were (Q, p<0.05; L, p<0.05) increased with infusion SSP. Total tract digestion of DM, OM and NDF were linearly (p<0.01) and quadratically (p<0.01) increased by infusing SSP. The flow of total amino acids (AA), essential amino acids (EAA) and individual amino acids were linearly (p<0.01) and quadratically (p<0.01) increased with infusion SSP. The digestibility of Lysine was quadratically (p = 0.033) increased and apparent degradability of Arginine was linearly (p = 0.032) and quadratically (p = 0.042) increased with infusion SSP. The results indicated that infusion SSP could improve nutrient digestion, ruminal fermentation and AA availability.


Severe atopic dermatitis and transient hypogammaglobulinemia in children.
We sought to describe the clinical outcomes of eight pediatric patients diagnosed with atopic dermatitis (AD) and hypogammaglobulinemia through retrospective review of medical records. All patients presented with severe facial AD. The mean and median ages of diagnosis of hypogammaglobulinemia were 6.2 months and 6.5 months, respectively, with a mean immunoglobulin G (IgG) level of 156 mg/dL. Seven of the eight patients identified in our search demonstrated simultaneous improvement in AD and serum IgG levels within 2 years of initial presentation, suggesting a diagnosis of transient hypogammaglobulinemia. The remaining patient demonstrated normalization by age 6, but no IgG levels had been measured between initial presentation and age 6. The five patients who were tested for specific antibody response to tetanus and Haemophilus influenzae type b vaccination all produced protective responses. All eight patients initially presented with high serum IgE levels. On initial evaluation, three patients had leukocytosis (white blood cell count >18,000 cells/μL), and six had peripheral blood eosinophilia. Three patients outgrew their AD by age 5, and five had clinically good to excellent control of their AD at their last visit, coincident with normalization of IgG levels. Although severe AD and immunoglobulin deficiency may rarely be associated with complex immunodeficiency disorders, our observations suggest that, with careful immunologic monitoring and diligent skin care, most children who present with severe AD and hypogammaglobulinemia exhibit improvement in dermatitis and serum IgG levels within 2 years of onset without major complications.