Effects of long-term desipramine administration on noradrenergic neurotransmission: electrophysiological studies in the rat brain.
The present studies were undertaken to assess in vivo the effects of repeated administration of the norepinephrine (NE) reuptake blocker desipramine (DMI) (5 or 10 mg/kg/day x 14 days) on five subsets of adrenoceptors and on the effectiveness of the NE reuptake process in the rat central nervous system. These receptors were: 1) the postsynaptic alpha-2 adrenoceptors which signal the suppressant effect of microiontophoretically applied NE on the firing activity of dorsal hippocampus pyramidal neurons; 2) the postsynaptic alpha-1 adrenoceptors which mediate the short-latency suppressant effect of endogenous NE, released by the electrical stimulation of the locus ceruleus (LC), on the firing activity of these same neurons; 3) the postsynaptic beta adrenoceptors which mediate the long-latency excitatory effect of LC stimulation on hippocampus pyramidal neurons; 4) the alpha-2 autoreceptors located on NE terminals in the hippocampus; and 5) the alpha-2 autoreceptors located on the cell body of LC NE neurons. The suppressant effect of microiontophoretic applications of NE on dorsal hippocampus pyramidal neurons was not changed by the long-term DMI treatments. The recovery time of the firing activity of these neurons after microiontophoretic application of NE, an index of the activity of the NE reuptake process, was the same in control and DMI-treated rats. However, the prolongation of this recovery time by the acute i.v. administration of DMI reached a plateau at a lower cumulative dose in long-term DMI-treated than in control rats. These results thus suggest that long-term administration of DMI did not affect the function of the NE transporter but reduced the number of DMI binding sites. The effectiveness of the 1 Hz stimulation of the LC in suppressing of the firing activity of pyramidal neurons was unchanged by either long-term DMI treatment (5 or 10 mg/kg/day), whereas that of the 5 Hz stimulation was enhanced only in rats on the 10-mg/kg regimen. This suggests that the function of terminal alpha-2 autoreceptors was decreased after this latter DMI regimen, because the reduction of the efficacy of the stimulation obtained by increasing the frequency from 1 to 5 Hz has been shown to be due to the activation of terminal alpha-2 adrenergic autoreceptors. In keeping with this interpretation, the reduction of the effectiveness of the stimulation of the LC by the systemic administration of the alpha-2 adrenoceptor agonist clonidine was attenuated in rats treated with the 10-mg/kg/day regimen of DMI, but not in those with the 5-mg/kg/day regimen.(ABSTRACT TRUNCATED AT 400 WORDS)


A Longitudinal Pathway for Clinical Educators: a Case Study.
This article was migrated. The article was marked as recommended. It is increasingly being recognised that medical education research and scholarship are essential if we are to provide innovative, research-informed teaching to medical students. Developing a discrete pathway for the professional development of clinical educators side by side with clinical training would enable such training to be offered to high caliber candidates. However, the challenges faced by clinical educators trying to deliver both the clinical and educational components of their training to a high standard must not be underestimated. In addition, employing institutions need to consider and agree criteria for career progression of these clinical academics. Identifying a mentor who can guide professional development is crucial, particularly in the early stages.


Interindividual variability of carboxymethylenebutenolidase homolog, a novel olmesartan medoxomil hydrolase, in the human liver and intestine.
Olmesartan medoxomil (OM) is a prodrug-type angiotensin II type 1 receptor antagonist. OM is rapidly converted into its active metabolite olmesartan by multiple hydrolases in humans, and we recently identified carboxymethylenebutenolidase homolog (CMBL) as one of the OM bioactivating hydrolases. In the present study, we further investigated the interindividual variability of mRNA and protein expression of CMBL and OM-hydrolase activity using 40 individual human liver and 30 intestinal specimens. In the intestinal samples, OM-hydrolase activity strongly correlated with the CMBL protein expression, clearly indicating that CMBL is a major contributor to the prodrug bioactivation in human intestine. The protein and activity were highly distributed in the proximal region (duodenum and jejunum) and decreased to the distal region of the intestine. Although there was high interindividual variability (16-fold) in both the protein and activity in the intestinal segments from the duodenum to colon, the interindividual variability in the duodenum and jejunum was relatively small (3.0- and 2.4-fold, respectively). In the liver samples, the interindividual variability in the protein and activity was 4.1- and 6.8-fold, respectively. No sex differences in the protein and activity were shown in the human liver or intestine. A genetically engineered Y155C mutant of CMBL, which was caused by a single nucleotide polymorphism rs35489000, showed significantly lower OM-hydrolase activity than the wild-type protein although no minor allele was genotyped in the 40 individual liver specimens.


Minor diterpene glycosides from the leaves of Stevia rebaudiana.
Two new diterpene glycosides in addition to five known glycosides have been isolated from a commercial extract of the leaves of Stevia rebaudiana. Compound 1 (rebaudioside KA) was shown to be 13-[(O-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid 2-O-β-d-glucopyranosyl-β-d-glucopyranosyl ester and compound 2, 12-α-[(2-O-β-d-glucopyranosyl-β-d-glucopyranosyl)oxy]ent-kaur-16-en-19-oic acid β-d-glucopyranosyl ester. Five additional known compounds were identified, rebaudioside E, rebaudioside M, rebaudioside N, rebaudioside O, and stevioside, respectively. Enzymatic hydrolysis of stevioside afforded the known ent-kaurane aglycone 13-hydroxy-ent-kaur-16-en-19-oic acid (steviol) (3). The isolated metabolite 1 possesses the ent-kaurane aglycone steviol (3), while compound 2 represents the first example of the isomeric diterpene 12-α-hydroxy-ent-kaur-16-en-19-oic acid existing as a glycoside in S. rebaudiana. The structures of the isolated metabolites 1 and 2 were determined based on comprehensive 1D- and 2D-NMR (COSY, HSQC, and HMBC) studies. A high-quality crystal of compound 3 has formed, which allowed the acquisition of X-ray diffraction data that confirmed its structure. The structural similarities between the new metabolites and the commercially available stevioside sweeteners suggest the newly isolated metabolites should be examined for their organoleptic properties. Accordingly rebaudiosides E, M, N, O, and KA have been isolated in greater than gram quantities.


Animals and alternatives: societal expectations and scientific need.
As Russell and Burch suggested more than 40 years ago, the most humane science is the best science. The path ahead is clear: pain and distress must be eliminated in animal experiments or reduced to an absolute minimum, and, as scientists, we must use the most humane approaches in our research. To accomplish the best science, we must train those who come after us in the principles and practice of humane science.


The effect of chronic retrovirus infection and immune dysfunction on the P-450-mediated activation of acetaminophen in mouse liver microsomes.
Acute viral infection has long been recognized to down-regulate cytochrome P-450 enzymes and subsequently to result in changes in the pharmacological and toxicological responses to xenobiotics. In our previous research, chronic retrovirus infection induced by inoculating a susceptible strain of mice with LP-BM5 murine leukaemia virus (MuLV) was found to suppress acetaminophen (APAP) induced liver injury. In the present study, we aimed to examine the influence of chronic retrovirus infection and its associated immune dysfunction on the activities of a number of cytochrome P-450 isozymes and the P-450-mediated activation of APAP in mouse liver microsomes. Liver microsomes prepared from female C57BL/6 mice at 8 and 16 weeks after LP-BM5 MuLV inoculation as well as from age-matched controls were used in the study. The catalytic activities of the cytochrome P-450 isozymes 1A family and 2E1, catalysts for the activation of APAP, were measured in different microsomal preparations using O-dealkylation of alkoxyresorufin homologues and oxidation of p-nitrophenol, respectively, as the metabolic markers. The formation of the reactive APAP metabolite trapped as glutathione conjugate in the microsomal preparations was also determined. We demonstrated that there were variable changes in total hepatic P-450 levels and in the activities of a number of P-450 isozymes in animals with chronic retrovirus infection and immune dysfunction. Such changes seemed to be dependent on the stage of the disease and to have resulted in increases or minimal changes in the rate of APAP activation in hepatic microsomes collected from this animal model. This suggests that the P-450-mediated activation of APAP was not down-regulated in animals with chronic retrovirus infection. Enhanced elimination of APAP by detoxification metabolic pathways is more likely to be responsible for the increased resistance to APAP-induced hepatotoxicity observed in our previous research in animals with chronic retrovirus infection.


Me, myself, and my muscles: associations between narcissism and muscle dysmorphia.
The conceptualisation and measurement of the associations between narcissism and muscle dysmorphia (MD) are poorly understood in the literature. It is recognised that narcissism features two broad dimensions: vulnerable and grandiose. Studying both narcissism dimensions is important given their differences in etiology and health-related outcomes. Therefore, the aim of this study was to investigate the associations between MD, vulnerable and grandiose narcissism. A total of 336 participants (48% male; Mage = 26.4 years) completed measures of MD and narcissism. Hierarchical regression analysis revealed that vulnerable narcissism was significantly, positively associated with MD, whilst grandiose narcissism was not. Practitioners should be aware that vulnerable narcissism may be a potential risk factor of MD.


Non-ribosomal factors in ribosome subunit assembly are emerging targets for new antibacterial drugs.
It is becoming increasingly clear that bacterial ribosome assembly is catalyzed by a variety of non-ribosomal factors. Newly characterized factors in bacterial ribosome biogenesis are broadly conserved and often indispensable proteins that can be classified either as chaperones facilitating assembly, or enzymes with ribosomal RNA- and ribosomal protein-modifying functions. Accumulating evidence indicates that the proteins Era, Obg, YjeQ, YlqF and RimM are chaperones which may be crucial to bacterial ribosome assembly, and therefore represent novel targets for modern antibacterial drug discovery. Ongoing work aimed at understanding ribosome biogenesis is expected to continue to yield additional factors crucial to this process, and provide new targets with drug discovery potential.


Conspecific olfactory preferences and interspecific divergence in odor cues in a chickadee hybrid zone.
Understanding how mating cues promote reproductive isolation upon secondary contact is important in describing the speciation process in animals. Divergent chemical cues have been shown to act in reproductive isolation across many animal taxa. However, such cues have been overlooked in avian speciation, particularly in passerines, in favor of more traditional signals such as song and plumage. Here, we aim to test the potential for odor to act as a mate choice cue, and therefore contribute to premating reproductive isolation between the black-capped (Poecile atricapillus) and Carolina chickadee (P. carolinensis) in eastern Pennsylvania hybrid zone populations. Using gas chromatography-mass spectrometry, we document significant species differences in uropygial gland oil chemistry, especially in the ratio of ester to nonester compounds. We also show significant preferences for conspecific over heterospecific odor cues in wild chickadees using a Y-maze design. Our results suggest that odor may be an overlooked but important mating cue in these chickadees, potentially promoting premating reproductive isolation. We further discuss several promising avenues for future research in songbird olfactory communication and speciation.


Protective role of ginger on lead induced derangement in plasma testosterone and luteinizing hormone levels of male Sprague Dawley rats.
Lead is one of the most serious environmental threats to human health especially in developing countries. It damages multiple body systems including the reproductive system. Ginger's antioxidant and androgenic activity is reported in multiple animal studies. The aim of this study was to investigate the ameliorative effect of Zingiber officinale (ginger) on lead induced derangement in plasma testosterone and luteinizing hormone (LH) levels of male rats. Sixty adult male Sprague Dawley rats were used in this study in four groups. Group A served as normal control, Group B received 0.3% lead acetate in drinking water, Group C and group D received supplementary 0.5 and 1 gm/Kg bodyweight of ginger respectively along with lead acetate in drinking water. Five rats from each group were sacrificed at the end of 2nd, 4th and 6th weeks. Serum testosterone and LH levels were analysed using ELISA technique. After co administration with different doses of ginger, serum testosterone level which was significantly decreased in lead treated group, showed a significant rise as compared to lead treated group. LH levels which had exhibited no significant change by lead treatment, after co administration with different doses of ginger, again showed no significant change. Oral administration of ginger ameliorated lead induced testicular toxicity in male rats by increasing serum testosterone level at all durations which might be a product of both its androgenic and antioxidant properties.


Photo-catalytic oxidation of cyclohexane over TiO2: a novel interpretation of temperature dependent performance.
The rate of cyclohexane photo-catalytic oxidation to cyclohexanone over anatase TiO(2) was studied at temperatures between 23 and 60 °C by in situ ATR-FTIR spectroscopy, and the kinetic parameters were estimated using a microkinetic model. At low temperatures, surface cyclohexanone formation is limited by cyclohexane adsorption due to unfavorable desorption of H(2)O, rather than previously proposed slow desorption of the product cyclohexanone. Up to 50 °C, the activation energy for photocatalytic cyclohexanone formation is zero, while carboxylates are formed with an activation energy of 18.4 ± 3.3 kJ mol(-1). Above 50 °C, significant (thermal) oxidation of cyclohexanone contributes to carboxylate formation. The irreversibly adsorbed carboxylates lead to deactivation of the catalyst, and are most likely the predominant cause of the non-Arrhenius behavior at relatively high reaction temperatures, rather than cyclohexane adsorption limitations. The results imply that elevating the reaction temperature of photocatalytic cyclohexane oxidation reduces selectivity, and is not a means to suppress catalyst deactivation.


[Antioxidant treatment with N-acetylcysteine and vitamin C in patients with chronic bronchitis].
N-acetylcysteine (NAC) is known to have direct antioxidant properties due to its thiol-group on the one hand as well as indirect antioxidant capacity as cysteine-donor to cellular glutathione-synthesis on the other hand. Therefore NAC appears to be attractive in antioxidant therapy of inflammatory disorders of the lung. This study aimed to investigate the use of antioxidant therapy with NAC in chronic bronchitis. In this randomized, double-blinded and placebo-controlled trial 100 patients divided into four groups were observed over a period of 3 months. The treatment consisted of either 600 mg NAC bid, 500 mg Vitamin C bid, a combination of those two substances using the same dosage or a placebo-preparation. The release of reactive oxygen species from isolated neutrophilic granulocytes and mononuclear cells was quantified before treatment and after three months using chemiluminescence (primary outcome parameter). Furthermore cellular glutathione content of these inflammatory cells was quantified spectrometrically. In addition, leukocyte-count and erythrocyte sedimentation rate as well as spirometry and a standardized symptom-based questionnaire were used to monitor the clinical efficacy. None of the above substances were able to significantly reduce the release of reactive oxygen species from the examined population of inflammatory cells. They also failed to increase intracellular glutathione-levels. Concordantly, no changes in spirometry and the results of the symptom-based questionnaire were found. In summary NAC, Vitamin C and the NAC/ Vitamin C-combination did neither enhance antioxidant protection in the blood nor is it of any clinical benefit in chronic bronchitis. Possible reasons may be a lack of antioxidant deficiency in these patients and negative feedback mechanisms of the glutathione-system.


A systems biology approach to construct the gene regulatory network of systemic inflammation via microarray and databases mining.
Inflammation is a hallmark of many human diseases. Elucidating the mechanisms underlying systemic inflammation has long been an important topic in basic and clinical research. When primary pathogenetic events remains unclear due to its immense complexity, construction and analysis of the gene regulatory network of inflammation at times becomes the best way to understand the detrimental effects of disease. However, it is difficult to recognize and evaluate relevant biological processes from the huge quantities of experimental data. It is hence appealing to find an algorithm which can generate a gene regulatory network of systemic inflammation from high-throughput genomic studies of human diseases. Such network will be essential for us to extract valuable information from the complex and chaotic network under diseased conditions. In this study, we construct a gene regulatory network of inflammation using data extracted from the Ensembl and JASPAR databases. We also integrate and apply a number of systematic algorithms like cross correlation threshold, maximum likelihood estimation method and Akaike Information Criterion (AIC) on time-lapsed microarray data to refine the genome-wide transcriptional regulatory network in response to bacterial endotoxins in the context of dynamic activated genes, which are regulated by transcription factors (TFs) such as NF-kappaB. This systematic approach is used to investigate the stochastic interaction represented by the dynamic leukocyte gene expression profiles of human subject exposed to an inflammatory stimulus (bacterial endotoxin). Based on the kinetic parameters of the dynamic gene regulatory network, we identify important properties (such as susceptibility to infection) of the immune system, which may be useful for translational research. Finally, robustness of the inflammatory gene network is also inferred by analyzing the hubs and "weak ties" structures of the gene network. In this study, Data mining and dynamic network analyses were integrated to examine the gene regulatory network in the inflammatory response system. Compared with previous methodologies reported in the literatures, the proposed gene network perturbation method has shown a great improvement in analyzing the systemic inflammation.


Dietary fiber and satiety: the effects of oats on satiety.
This review examines the effect of β-glucan, the viscous soluble fiber in oats, on satiety. A literature search for studies that examined delivery of the fiber in whole foods or as an extract was conducted. Viscosity interferes with the peristaltic mixing process in the small intestine to impede digestion and absorption of nutrients, which precipitates satiety signals. From measurements of the physicochemical and rheological properties of β-glucan, it appears that viscosity plays a key role in modulating satiety. However, the lack of standardized methods to measure viscosity and the inherent nature of appetite make it difficult to pinpoint the reasons for inconsistent results of the effects of oats on satiety. Nevertheless, the majority of the evidence suggests that oat β-glucan has a positive effect on perceptions of satiety.


Enhancement of bone-titanium integration profile with UV-photofunctionalized titanium in a gap healing model.
In this study, we tested the potential of UV-photofunctionalized titanium surfaces to overcome compromised bone-titanium integration in a gap healing model. Titanium in rod and disk forms was acid etched and then stored for 4 weeks under dark ambient conditions. Titanium rods with and without UV pretreatment were placed into a rat femur with (contact healing) or without (gap healing) contact with the innate cortical bone. The titanium implants were subjected to a biomechanical push-in test, micro-CT bone morphometry, and surface elemental analysis after 2 weeks of healing. The strength of bone-titanium integration in the gap healing model was one-third of that in the contact healing model. However, UV-treated implants in the gap healing condition produced a strength of bone-titanium integration equivalent to that of untreated implants in the contact healing condition. Bone volume around UV-treated implants was 2- to 3-fold greater than that around the untreated implants in the gap healing model. A bone generation profile drawn along the long axis of the implant exhibited greater contrast between the untreated and UV-treated surfaces in the cortical area than in the bone marrow area. The bone tissue formed on UV-treated implants showed a higher Ca/P ratio than that formed on untreated titanium. The rate of cell proliferation, alkaline phosphatase activity, and calcium deposition in femoral periosteal cells and in bone marrow-derived osteoblasts were greater in cultures on UV-treated titanium disks than in cultures on untreated disks. The UV-enhanced function in periosteal cells was more pronounced when they were co-cultured with bone marrow-derived osteoblasts, indicating a synergistic effect of UV-treated titanium with biological signals from bone marrow-derived osteoblasts. Within the limitation of the model used in this study, UV-photofunctionalized titanium surfaces may overcome the challenging condition of bone-titanium integration without cortical bone support. UV treatment of implants induced marked improvements in the behavior of bone formation and quantity and quality of bone tissue around the implants. These effects may be related to the promoted function of both periosteum- and bone marrow-derived osteogenic cells at the local level around UV-treated titanium surfaces.


Lysosomal membrane protein composition, acidic pH and sterol content are regulated via a light-dependent pathway in metazoan cells.
In metazoans, lysosomes are characterized by a unique tubular morphology, acidic pH, and specific membrane protein (LAMP) and lipid (cholesterol) composition as well as a soluble protein (hydrolases) composition. Here we show that perturbation to the eye-color gene, light, results in impaired lysosomal acidification, sterol accumulation, altered endosomal morphology as well as compromised lysosomal degradation. We find that Drosophila homologue of Vps41, Light, regulates the fusion of a specific subset of biosynthetic carriers containing characteristic endolysosomal membrane proteins, LAMP1, V0-ATPase and the cholesterol transport protein, NPC1, with the endolysosomal system, and is then required for the morphological progression of the multivesicular endosome. Inhibition of Light results in accumulation of biosynthetic transport intermediates that contain these membrane cargoes, whereas under similar conditions, endosomal delivery of soluble hydrolases, previously shown to be mediated by Dor, the Drosophila homologue of Vps18, is not affected. Unlike Dor, Light is recruited to endosomes in a PI3P-sensitive fashion wherein it facilitates fusion of these biosynthetic cargoes with the endosomes. Depletion of the mammalian counterpart of Light, hVps41, in a human cell line also inhibits delivery of hLAMP to endosomes, suggesting an evolutionarily conserved pathway in metazoa.


[Use of chromatographic-mass spectrometers in diagnosis].
Conditions for extraction, desorption and concentration of specific substances, such as metabolites, and characteristics of their gas-chromatographic division and mass-spectrometric identification have been established. A library for mass-spectra of specific metabolite compounds characteristic of various diseases has been set up. Computer programs for metabolite analysis and for diagnosis were modified. An experimental introduction of chromatographic-mass-spectrometric diagnosis into practice in a number of hospitals and research institutions for prophylactic medical examination of people will make it possible to quickly accumulate statistic data and create the necessary data bank.


A Bacteroidetes locus dedicated to fungal 1,6-β-glucan degradation: Unique substrate conformation drives specificity of the key endo-1,6-β-glucanase.
Glycans are major nutrients available to the human gut microbiota. The Bacteroides are generalist glycan degraders, and this function is mediated largely by polysaccharide utilization loci (PULs). The genomes of several Bacteroides species contain a PUL, PUL1,6-β-glucan, that was predicted to target mixed linked plant 1,3;1,4-β-glucans. To test this hypothesis we characterized the proteins encoded by this locus in Bacteroides thetaiotaomicron, a member of the human gut microbiota. We show here that PUL1,6-β-glucan does not orchestrate the degradation of a plant polysaccharide but targets a fungal cell wall glycan, 1,6-β-glucan, which is a growth substrate for the bacterium. The locus is up-regulated by 1,6-β-glucan and encodes two enzymes, a surface endo-1,6-β-glucanase, BT3312, and a periplasmic β-glucosidase that targets primarily 1,6-β-glucans. The non-catalytic proteins encoded by PUL1,6-β-glucan target 1,6-β-glucans and comprise a surface glycan-binding protein and a SusD homologue that delivers glycans to the outer membrane transporter. We identified the central role of the endo-1,6-β-glucanase in 1,6-β-glucan depolymerization by deleting bt3312, which prevented the growth of B. thetaiotaomicron on 1,6-β-glucan. The crystal structure of BT3312 in complex with β-glucosyl-1,6-deoxynojirimycin revealed a TIM barrel catalytic domain that contains a deep substrate-binding cleft tailored to accommodate the hook-like structure adopted by 1,6-β-glucan. Specificity is driven by the complementarity of the enzyme active site cleft and the conformation of the substrate. We also noted that PUL1,6-β-glucan is syntenic to many PULs from other Bacteroidetes, suggesting that utilization of yeast and fungal cell wall 1,6-β-glucans is a widespread adaptation within the human microbiota.


Clinical and pathological characteristics of early-onset colorectal cancer in South Korea.
Early-onset colorectal cancer (EOCRC) may differ by race and ethnicity, and recently South Korea has witnessed a surge in cases. We aimed to evaluate the clinical and pathological features of patients with EOCRC, and to determine the predictors of overall survival. In this retrospective study, EOCRC was defined as CRC diagnosed in patients aged < 50 years, and late-onset CRC was defined as CRC diagnosed in those over 75 years of age. The clinical and pathological characteristics of patients with EOCRC were compared with late-onset CRC. We also used multivariable Cox proportional hazard models to find predictors of overall survival in patients with EOCRC. The proportion of early-onset CRC was 9.1% of 518 patients with CRC, and the clinical and pathological characteristics were similar between early-onset (n = 47) and late-onset CRC (n = 134). However, EOCRC had a preponderance for distal tumor location (70.2% vs. 50.7%, P = 0.02) and T1-2 stage disease (23.4% vs. 11.2%, P = 0.04), compared with those of late-onset CRC. Using multivariable Cox proportional hazard models, only vascular invasion (hazard ratio = 8.75, 95% confidence interval 1.139‒67.197) was found to be a risk factor for overall survival (P = 0.04) for patients with CRC. EOCRC had preponderance for distal tumor location and early T-stage disease, compared with late-onset CRC. Considering the increasing incidence of EOCRC, more studies on clinical and pathological characteristics of EOCRC may be warranted.


Blackbody spectrum revisited in the near field.
We report local spectra of the near-field thermal emission recorded by a Fourier transform infrared spectrometer, using a tungsten tip as a local scatterer coupling the near-field thermal emission to the far field. Spectra recorded on silicon carbide and silicon dioxide exhibit temporal coherence due to thermally excited surface waves. Finally, we evaluate the ability of this spectroscopy to probe the frequency dependence of the electromagnetic local density of states.


Pelvic Discontinuity Associated With Total Hip Arthroplasty: Evaluation and Management.
Pelvic discontinuity is a challenging complication encountered during revision total hip arthroplasty. Pelvic discontinuity is defined as a separation of the ilium superiorly from the ischiopubic segment inferiorly and is typically a chronic condition in failed total hip arthroplasties in the setting of bone loss. After a history and a physical examination have been completed and infection has been ruled out, appropriate imaging must be obtained, including plain hip radiographs, oblique Judet radiographs, and often a CT scan. The main management options are a hemispheric acetabular component with posterior column plating, a cup-cage construct, pelvic distraction, and a custom triflange construct. The techniques have unique pros and cons, but the goals are to obtain stable and durable acetabular component fixation and a healed or unitized pelvis while minimizing complications.


A spatio-temporal atlas of the developing fetal brain with spina bifida aperta.
Background: Spina bifida aperta (SBA) is a birth defect associated with severe anatomical changes in the developing fetal brain. Brain magnetic resonance imaging (MRI) atlases are popular tools for studying neuropathology in the brain anatomy, but previous fetal brain MRI atlases have focused on the normal fetal brain. We aimed to develop a spatio-temporal fetal brain MRI atlas for SBA. Methods: We developed a semi-automatic computational method to compute the first spatio-temporal fetal brain MRI atlas for SBA. We used 90 MRIs of fetuses with SBA with gestational ages ranging from 21 to 35 weeks. Isotropic and motion-free 3D reconstructed MRIs were obtained for all the examinations. We propose a protocol for the annotation of anatomical landmarks in brain 3D MRI of fetuses with SBA with the aim of making spatial alignment of abnormal fetal brain MRIs more robust. In addition, we propose a weighted generalized Procrustes method based on the anatomical landmarks for the initialization of the atlas. The proposed weighted generalized Procrustes can handle temporal regularization and missing annotations. After initialization, the atlas is refined iteratively using non-linear image registration based on the image intensity and the anatomical land-marks. A semi-automatic method is used to obtain a parcellation of our fetal brain atlas into eight tissue types: white matter, ventricular system, cerebellum, extra-axial cerebrospinal fluid, cortical gray matter, deep gray matter, brainstem, and corpus callosum. Results: An intra-rater variability analysis suggests that the seven anatomical land-marks are sufficiently reliable. We find that the proposed atlas outperforms a normal fetal brain atlas for the automatic segmentation of brain 3D MRI of fetuses with SBA. Conclusions: We make publicly available a spatio-temporal fetal brain MRI atlas for SBA, available here: https://doi.org/10.7303/syn25887675. This atlas can support future research on automatic segmentation methods for brain 3D MRI of fetuses with SBA.


Clinical Gait Analysis on a Patient Undergoing Surgical Correction of Kyphosis from Severe Ankylosing Spondylitis.
Ankylosing spondylitis (AS) is a chronic inflammatory disease primarily affecting the axial skeleton, including the sacroiliac joints, costovertebral joints, and the spine. Patients with AS found to have an alter gait pattern. The purpose of this study was to investigate biomechanical alterations in gait after surgical correction in a patient with severe kyphosis from AS. A case report in controlled laboratory study, a pretest-posttest design. A 20 year-old male presented with severe sagittal imbalance and inability to stand erect due to AS. He presented with thoracic kyphosis of 70°, lumbar kyphosis of 25°, and pelvic incidence of 43°. The patient had a complex spinal reconstruction with 84° of sagittal correction, normalizing his sagittal alignment. Gait analysis was performed the day before surgery and one month post surgery, including three-dimensional kinematics, ground reaction forces, and electromyography outcomes. Normalization of spinal alignment minimally increased walking speed and cadence. Lower extremity ranges of motion angles increased, but were not symmetrical even one month post surgery. Postoperatively, trunk flexion, neck extension and head orientation angles decreased compared with preoperative values, but was not symmetrical even one month post surgery. The trunk muscles were activated earlier in the post surgery condition compared to the pre surgery condition while lower extremity muscles presented later muscle activation. Surgical correction of spinal alignment improved spine function and efficiency. Changes in gait abnormality parameters observed imply that the patient used less energy to ambulate after surgery than before surgery. Although pre-surgery data showed compensation in the spine kinematics, post-surgery data supported significant changes in the spine and the lower extremity values. Formal gait and motion analysis can provide a method to assess the impact of severe spinal deformity on function and changes after treatment.


Outcomes after pneumonectomy for benign disease: the impact of urgent resection.
Pneumonectomy for benign disease is often complicated by inflammatory processes that obscure operative planes. We reviewed our experience to evaluate the impact of requiring urgent or emergent pneumonectomy on outcomes. All pneumonectomies for benign conditions from 1997 to 2012 at a single institution were retrospectively reviewed. Mortality was assessed using multivariable logistic regression that included laterality, age, and surgery status, which was emergent if performed within 24 hours of initial evaluation, urgent if performed after 24 hours but within the same hospital stay, and otherwise elective. Among 42 pneumonectomies, completion pneumonectomy after previous ipsilateral lung resection was performed in 14 patients (33%). Resection was elective in 22 patients (52%), urgent in 12 (28%), and emergent in 8 (19%). The most common indication was for necrotic lung (n = 12; 29%). Muscle flaps were used in 26 patients (62%). Perioperative mortality for the entire cohort was 29% (n = 12) and was significantly higher when surgery was urgent (5 of 12; 42%) or emergent (5 of 8; 62.5%) compared with elective (2 of 22; 9.1%) (p = 0.03). Requiring urgent or emergent surgery remained a significant predictor of mortality in multivariable analysis (odds ratio 10.4, p = 0.01). Pneumonectomy for benign disease has significant risk for mortality, particularly when not performed electively. Although surgery cannot be planned in the setting of trauma or some situations of acute infection, patients known to have conditions that are likely to require pneumonectomy should be considered for surgery earlier in their disease course, before developing an acute problem that requires urgent or emergent resection.


Investigation of polyomaviruses replication in pediatric patients with nephropathy receiving rituximab.
Rituximab is a chimeric monoclonal antibody reacting with the CD20 antigen on B cells. It has been proposed as treatment for the idiopathic nephrotic syndrome, recurrent idiopathic nephropathy, and focal segmental glomerulosclerosis refractory to steroids. Rituximab influences T-cell immunity and may predispose the patients to opportunistic infections, such as progressive multifocal leukoencephalopathy caused by the polyomavirus JC (JCV). The risk of latent viruses infections/reactivations in pediatric patients receiving monoclonal antibodies is not well known yet. In this longitudinal 6-month study, the effects of rituximab on JCV and BK virus (BKV) replication have been investigated. Blood, serum, and urine samples have been collected monthly from 11 pediatric patients (mean age: 11 years) with the idiopathic nephrotic syndrome and recurrent idiopathic nephropathy, under rituximab therapy. JCV and BKV real-time PCRs and sequencing of the viral protein 1 and the non-coding control region have been conducted. The same investigations have been undertaken on samples collected from eight pediatric patients (controls, mean age: 6 years), with idiopathic nephrotic syndrome or focal segmental glomerulosclerosis, treated with conventional chemotherapy. JCV was detected in the urine of one patient (9%), and one control (12.5%); BKV was found in the urine of 7/11 patients (63.6%) and 2/8 controls (25%) and in blood samples from four patients. No significant difference was found in the mean viral loads and in the viral molecular characterizations between the two groups. The polyomaviruses replication was not associated with rituximab therapy in children.


The 150 most important questions in cancer research and clinical oncology series: questions 67-75 : Edited by Chinese Journal of Cancer.
Since the beginning of 2017, Chinese Journal of Cancer has published a series of important questions in cancer research and clinical oncology, which sparkle diverse thoughts, interesting communications, and potential collaborations among researchers all over the world. In this article, 9 more questions are presented as followed. Question 67. How could we overcome the resistance of hepatocellular carcinoma against chemotherapeutics? Question 68. Is pursuit of non-covalent small-molecule binders of RAS proteins viable as a strategy of cancer drug discovery? Question 69. In what oligomeric structures do RAS proteins signal? Question 70. How can we achieve non-invasive early detection and diagnosis of lung cancer? Question 71. Does genetic information influence the volatolome enabling diagnosis of lung cancer with genetic mutations via cell headspace or breath analysis? Question 72. Is heavy ion beam radiotherapy effective to kill cancer stem cells? Question 73. Is there any diversity among different types of cancer in terms of sensitivity to heavy ion beam radiotherapy? Question 74. Can targeted alpha-particle therapy augment the effect of carbon ion radiotherapy on malignancies? Question 75. How does chromosomal instability drive tumor progression?


Comparison of the health and lifestyle of general practitioners and teachers.
A total of 704 general practitioners and 588 teachers responded to a questionnaire about their health and lifestyle in 1991 (response rates 82% and 87%, respectively). The results for lifestyle measures were compared with those of a similar questionnaire completed by about half of each group two years before--there were no changes in the answers of either occupational group in the intervening two years. In 1991, 9% of general practitioners and 15% of teachers drank 22 units of alcohol per week or more; 13% of general practitioners and 23% of teachers reported troublesome depression and 31% of doctors and 37% of teachers excessive anxiety in the preceding 12 months. Teachers had more sickness absence, and significantly more experienced a need for daily alcohol and binge eating, and reported sleep difficulties, depression and anxiety than general practitioners. Self-medication among general practitioners was common and overall accounted for 83% of the medication taken by doctors. A follow-up survey of non-respondents found that only 11% of general practitioners and 11% of teachers indicated they had a health problem they wished to conceal or that they felt the questions were too intimate. General practitioners' lifestyle habits are better than those of teachers and published figures for the general population. The frequency of reported mental health problems in both professions gives cause for concern.


Porcine Knock-in Fibroblasts Expressing hDAF on α-1,3-Galactosyltransferase (GGTA1) Gene Locus.
The Galactose-α1,3-galactose (α1,3Gal) epitope is responsible for hyperacute rejection in pig-to-human xenotransplantation. Human decay-accelerating factor (hDAF) is a cell surface regulatory protein that serves as a complement inhibitor to protect self cells from complement attack. The generation of α1,3-galactosyltransferase (GGTA1) knock-out pigs expressing DAF is a necessary step for their use as organ donors for humans. In this study, we established GGTA1 knock-out cell lines expressing DAF from pig ear fibroblasts for somatic cell nuclear transfer. hDAF expression was detected in hDAF knock-in heterozygous cells, but not in normal pig cells. Expression of the GGTA1 gene was lower in the knock-in heterozygous cell line compared to the normal pig cell. Knock-in heterozygous cells afforded more effective protection against cytotoxicity with human serum than with GGTA1 knock-out heterozygous and control cells. These cell lines may be used in the production of GGTA1 knock-out and DAF expression pigs for xenotransplantation.


The microbiome of pancreatic cancer: from molecular diagnostics to new therapeutic approaches to overcome chemoresistance caused by metabolic inactivation of gemcitabine.
Introduction: Pancreatic cancer is a complex disease, with an extremely poor response to chemotherapy. Emerging evidence indicates that the tumor microenvironment (TME) might play an important role in mediating chemoresistance. Areas covered: The evaluated study by Geller and collaborators describes several bacterial species within pancreatic tumor tissues and TME and investigated their roles in gemcitabine chemoresistance. Intratumor bacteria express the enzyme cytidine deaminase (CDD), whose long form (CDDL) was shown to metabolize gemcitabine into its inactive metabolite. CDDL is mostly expressed by Gammaproteobacteria and this was among the most common species in pancreatic cancer tissues. Interestingly, mouse models of colorectal cancer injected with bacterial CDDL displayed a reduced response to gemcitabine, but this resistance was neutralized by the antibiotic ciprofloxacin. Expert Commentary: The increased knowledge on the microbiome in pancreatic tissues, as well as its role in chemoresistance, will provide innovative prognostic and therapeutic strategies.


Normal development of the gut-associated lymphoid tissue except Peyer's patch in MyD88-deficient mice.
MyD88 is a key adaptor molecule for signalling via Toll-like receptors (TLRs) and the response to gut commensal microbes. To investigate the role of TLRs/MyD88 pathway in the development of the gut-associated lymphoid tissue (GALT), we examined the development of Peyer's patches (PPs) and cryptopatch (CP), and also one of effector compartment, intraepithelial lymphocyte (IEL) in MyD88-/-, TLR2-/- and TLR4-/- mice. In MyD88-/- mice, the organogenesis of PPs was not disturbed. However, PPs in 2-week-old MyD88-/- mice were significantly smaller than those in MyD88+/- mice. Also, in 2-week-old TLR4-/-, but not TLR2-/- mice, PPs did not develop rapidly. The development of PPs in MyD88-/- and TLR4-/- mice was completely recovered in 10 weeks. PP cells from MyD88-/- mice showed significant decrease in proliferation when stimulated with lipopolysaccharide. The development of CP and IEL was also normal in 10-week-old MyD88-/- mice. These results suggest that the TLRs/MyD88 pathway might be involved in the development of PPs only at early postnatal stage, and TLRs/MyD88-independent signalling is critically involved in the development of GALT in adult mice.


Molecular recognition of methyl α-D-mannopyranoside by antifreeze (glyco)proteins.
Antifreeze proteins and glycoproteins [AF(G)Ps] have been well-known for more than three decades for their ability to inhibit the growth and recrystallization of ice through binding to specific ice crystal faces, and they show remarkable structural compatibility with specific ice crystal faces. Here, we show that the crystal growth faces of methyl α-D-mannopyranoside (MDM), a representative pyranose sugar, also show noteworthy structural compatibility with the known periodicities of AF(G)Ps. We selected fish AFGPs (AFGP8, AFGP1-5), and a beetle AFP (DAFP1) with increasing antifreeze activity as potential additives for controlling MDM crystal growth. Similar to their effects on ice growth, the AF(G)Ps can inhibit MDM crystal growth and recrystallization, and more significantly, the effectiveness for the AF(G)Ps are well correlated with their antifreeze activity. MDM crystals grown in the presence of AF(G)Ps are smaller and have better defined shapes and are of higher quality as indicated by single crystal X-ray diffraction and polarized microscopy than control crystals, but no new polymorphs of MDM were identified by single crystal X-ray diffraction, solid-state NMR, and attenuated total reflectance infrared spectroscopy. The observed changes in the average sizes of the MDM crystals can be related to the changes in the number of the MDM nuclei in the presence of the AF(G)Ps. The critical free energy change differences of the MDM nucleation in the absence and presence of the additives were calculated. These values are close to those of the ice nucleation in the presence of AF(G)Ps suggesting similar interactions are involved in the molecular recognition of MDM by the AF(G)Ps. To our knowledge this is the first report where AF(G)Ps have been used to control crystal growth of carbohydrates and on AFGPs controlling non-ice-like crystals. Our finding suggests MDM might be a possible alternative to ice for studying the detailed mechanism of AF(G)P-crystal interactions. The relationships between AF(G)Ps and carbohydrate binding proteins are also discussed. The structural compatibility between AF(G)Ps and growing crystal faces demonstrated herein adds to the repertoire of molecular recognition by AF(G)Ps, which may have potential applications in the sugar, food, pharmaceutical, and materials industries.


Imparting Immunomodulatory Activity to Scaffolds via Biotin-Avidin Interactions.
Biotin-avidin interactions have been explored for decades as a technique to functionalize biomaterials, as well as for in vivo targeting, but whether changes in these interactions can be leveraged for immunomodulation remain unknown. The goal of this study was to investigate how biotin density and avidin variant can be used to deliver the immunomodulatory cytokine, interleukin 4 (IL4), from a porous gelatin scaffold, Gelfoam, to primary human macrophages in vitro. Here, we demonstrate that the degree of scaffold biotinylation controlled the binding of two different avidin variants, streptavidin and CaptAvidin. Biotinylated scaffolds were also loaded with streptavidin and biotinylated IL4 under flow, suggesting a potential use for targeting this biomaterial in vivo. While biotin-avidin interactions did not appear to influence the protein release in this system, increasing degrees of biotinylation did lead to increased M2-like polarization of primary human macrophages over time in vitro, highlighting the capability to leverage biotin-avidin interactions to modulate the macrophage phenotype. These results demonstrate a versatile and modular strategy to impart immunomodulatory activity to biomaterials.


Herpes simplex virus type 1-derived recombinant and amplicon vectors.
Herpes simplex virus type 1 (HSV-1) is a human pathogen whose lifestyle is based on a long-term dual interaction with the infected host, being able to establish both lytic and latent infections. The virus genome is a 153 kbp double-stranded DNA molecule encoding more than 80 genes. The interest of HSV-1 as gene transfer vector stems from its ability to infect many different cell types, both quiescent and proliferating cells, the very high packaging capacity of the virus capsid, the outstanding neurotropic adaptations that this virus has evolved, and the fact that it never integrates into the cellular chromosomes, thus avoiding the risk of insertional mutagenesis. Two types of vectors can be derived from HSV-1, recombinant vectors and amplicon vectors, and different methodologies have been developed to prepare large stocks of each type of vector. This chapter summarizes (1) the two approaches most commonly used to prepare recombinant vectors through homologous recombination, either in eukaryotic cells or in bacteria, and (2) the two methodologies currently used to generate helper-free amplicon vectors, either using a bacterial artificial chromosome (BAC)-based approach or a Cre/loxP site-specific recombination strategy.


Death, child abuse, and adverse neurological outcome of infants after an apparent life-threatening event.
Apparent life-threatening events in infants constitute a significant challenge for health care providers. Apparent life-threatening event evaluation and management are poorly defined, and outcomes have not been clearly determined. Our objectives were to characterize short- and long-term risks for death, child abuse, and abnormal neurological outcomes of infants after an apparent life-threatening event and to identify clinical features that are predictive of these outcomes. We collected data from infants ages birth to 12 months of age who were hospitalized after an apparent life-threatening event during a 5-year time period. Patients were evaluated for subsequent death, child abuse, or adverse neurological outcome (chronic epilepsy or developmental delay). A total of 471 patients met inclusion criteria and were followed an average of 5.1 years. Two patients died after developing chronic epilepsy and severe developmental delay. Fifty-four (11%) patients were diagnosed as being a victim of child abuse, but only 2 were identified at initial presentation. There were 23 (4.9%) patients with adverse neurological outcomes, including 17 (3.6%) with chronic epilepsy and 14 (3.0%) with developmental delay. Of those who developed chronic epilepsy, 71% returned within 1 month of the initial apparent life-threatening event with a second event. Neurological evaluation at the time of the apparent life-threatening event had low yield for predicting those who would develop chronic epilepsy. Infants who suffer an apparent life-threatening event are at risk for subsequent child abuse and adverse neurological outcomes. Deaths were uncommon and only occurred in the setting of severe developmental delay and seizure disorders. Neurological evaluation during hospitalization for a first apparent life-threatening event is of low yield, but close follow-up is essential.


The realistic domain structure of as-synthesized graphene oxide from ultrafast spectroscopy.
Graphene oxide (GO) is an attractive alternative for large-scale production of graphene, but its general structure is still under debate due to its complicated nonstoichiometric nature. Here we perform a set of femtosecond pump-probe experiments on as-synthesized GO to extrapolate structural information in situ. Remarkably, it is observed that, in these highly oxidized GO samples, the ultrafast graphene-like dynamics intrinsic to pristine graphene is completely dominant over a wide energy region and can be modified by the localized impurity states and the electron-phonon coupling under certain conditions. These observations, combined with the X-ray photoelectron spectroscopy analysis and control experiments, lead to an important conclusion that GO consists of two types of domain, namely the carbon-rich graphene-like domain and the oxygen-rich domain. This study creates a new understanding of the realistic domain structure and properties of as-synthesized GO, offering useful guidance for future applications based on chemically modified/functionalized graphenes.


Evaluation of Bt resistance in Helicoverpa zea (Lepidoptera: Noctuidae) strains using various Bt cotton plant tissues.
Diet-overlay bioassays suggest that Helicoverpa zea (Lepidoptera: Noctuidae) field populations have developed resistance to some of the Bt insecticidal proteins that are constituents of the pyramids expressed in the second and third generation Bt cotton technologies. Unfortunately, these bioassays are not always a reliable indicator for how a seemingly resistant population will perform in an actual cotton field, and thus, leaf tissue bioassays have been suggested as a method to better assess field performance. However, bollworm larvae typically prefer to feed on floral tissue rather than leaf tissue, and an alternative cotton structure type may be more ideal for use in plant tissue-based bioassays. A series of diet-overlay bioassays using Bt proteins and Bt cotton plant tissue were conducted with laboratory susceptible (Bz-SS) and resistant (Cry-RR, resistant to Cry1Ac and Cry2Ab) H. zea strains to determine if plant tissue overlays could detect resistance and which cotton plant structure type would be most ideal for use in bioassays. Results suggest that diet overlays using lyophilized plant tissue were able to detect resistance. Lyophilized tissue from white flowers was most ideal for use in bioassays, whereas tissue from non-Bt bolls and leaves affected larval health and behavior, confounding assay results. Overlays using white flower tissue could potentially be used to supplement Bt protein overlays and provide an improved assessment of larval performance on Bt cotton technologies. © 2021 Society of Chemical Industry.


Surface states in nearly modulated systems.
A Landau model is used to study the phase behavior of the surface layer for magnetic and cholesteric liquid-crystal systems that are at or near a Lifshitz point marking the boundary between modulated and homogeneous bulk phases. The model incorporates surface and bulk fields and includes a term in the free energy proportional to the square of the second derivative of the order parameter in addition to the usual term involving the square of the first derivative. In the limit of vanishing bulk field, three distinct types of surface ordering are possible: a wetting layer, a nonwet layer having a small deviation from bulk order, and a different nonwet layer with a large deviation from bulk order that decays nonmonotonically as the distance from the wall increases. In particular, the large deviation nonwet layer is a feature of systems at the Lifshitz point and also those systems having only homogeneous bulk phases.


Laparoscopic versus open ventral hernia repair.
Ventral hernia repair remains one of the most common operations performed by general surgeons. Despite the frequency with which this procedure is performed, there is little agreement and extensive controversy as to the cause of most of the hernias, or the ideal approach to repair these complicated problems. This article attempts to identify and provide some clarification of these controversial issues in abdominal wall reconstruction after ventral herniation based on the available literature.


Mice with induced pulmonary morbidities display severe lung inflammation and mortality following exposure to SARS-CoV-2.
Mice are normally unaffected by SARS coronavirus 2 (SARS-CoV-2) infection since the virus does not bind effectively to the murine version of the angiotensin-converting enzyme 2 (ACE2) receptor molecule. Here, we report that induced mild pulmonary morbidities rendered SARS-CoV-2-refractive CD-1 mice susceptible to this virus. Specifically, SARS-CoV-2 infection after application of low doses of the acute lung injury stimulants bleomycin or ricin caused severe disease in CD-1 mice, manifested by sustained body weight loss and mortality rates greater than 50%. Further studies revealed markedly higher levels of viral RNA in the lungs, heart, and serum of low-dose ricin-pretreated mice compared with non-pretreated mice. Furthermore, lung extracts prepared 2-3 days after viral infection contained subgenomic mRNA and virus particles capable of replication only when derived from the pretreated mice. The deleterious effects of SARS-CoV-2 infection were effectively alleviated by passive transfer of polyclonal or monoclonal antibodies generated against the SARS-CoV-2 receptor binding domain (RBD). Thus, viral cell entry in the sensitized mice seems to depend on viral RBD binding, albeit by a mechanism other than the canonical ACE2-mediated uptake route. This unique mode of viral entry, observed over a mildly injured tissue background, may contribute to the exacerbation of coronavirus disease 2019 (COVID-19) pathologies in patients with preexisting morbidities.


Multifaceted nutritional intervention among nursing-home residents has a positive influence on nutrition and function.
We tested the hypothesis that a multifaceted 11-wk intervention comprising nutrition, group exercise, and oral care would have a significant influence on nutrition and function in elderly (>or=65 y) nursing-home residents. The study was an 11-wk randomized controlled intervention study with nutrition (chocolate and homemade oral supplements), group exercise twice a week (45-60 min, moderate intensity), and oral care intervention one to two times a week, with the aim of improving nutritional status and function in elderly nursing-home residents. A follow-up visit was made 4 mo after the end of the intervention. Assessments were weight, body mass index, dietary intake, handgrip strength, Senior Fitness Test, Berg's Balance Scale, and the prevalence of plaque. A total of 121 subjects (61%) accepted the invitation and 62 were randomized to the intervention group. Six of these dropped out during the 11 wk. At the 4-mo follow-up there were 15 deaths in the intervention group and 8 in the control group. The nutrition and exercise were well tolerated. After 11 wk the change in percentage of weight (P = 0.005), percentage of body mass index (P = 0.003), energy intake (P = 0.084), protein intake (P = 0.012), and Berg's Balance Scale (P = 0.004) was higher in the intervention group than in the control group. In addition, the percentage of subjects whose functional tests improved was higher in the intervention group. Both groups lost the same percentage of weight after the intervention (P = 0.908). The total percentage of weight loss from baseline to follow-up was higher in the control group (P = 0.019). Oral care was not well accepted and the prevalence of plaque did not change. It is possible to improve nutrition and function in elderly nursing-home residents by means of a multifaceted intervention consisting of chocolate, homemade supplements, group exercise, and oral care.


Acute versus chronic supplementation of sodium citrate on 200 m performance in adolescent swimmers.
A double-blinded, placebo-controlled, cross-over design was used to investigate whether two different sodium citrate dihydrate (Na-CIT) supplementation protocols improve 200 m swimming performance in adolescent swimmers. Ten, male swimmers (14.9 ± 0.4 years of age; 63.5 ± 4 kg) performed four 200 m time trials with the following treatments: acute (ACU) supplementation (0.5 g kg(-1) administered 120 min pre-trial), acute placebo (PLC-A), chronic (CHR) supplementation (0.1 g∙kg(-1) for three days and 0.3 g kg(-1) on the forth day 120 min pre-trial), and chronic placebo (PLC-C). The order of the trials was randomized, with at least a six-day wash-out period between trials. Blood samples were collected by finger prick pre-ingestion, 100 min post-ingestion, and 3 min post-trial. Performance time, rate of perceived exertion, pH, base excess, bicarbonate and lactate concentration were measured. Post-ingestion bicarbonate and base excess were higher (P < 0.05) in both the ACU and CHR trials compared to placebo showing adequate pre-exercise alkalosis. However, performance time, rate of perceived exertion as well as post-trial pH and lactate concentration were not significantly different between trials. Further analysis revealed that five swimmers, identified as responders, improved their performance time by 1.03% (P < 0.05) and attained higher post-trial lactate concentrations in the ACU versus PLC-A trial (P < 0.05). They also had significantly higher post-trial lactate concentrations compared to the non-responders in the ACU and CHR trials. Acute supplementation of Na-CIT prior to 200 m swimming performance led to a modest time improvement and higher blood lactate concentrations in only half of the swimmers while the chronic Na-CIT supplementation did not provide any ergogenic effect in this group of adolescent swimmers. Clinicaltrials.gov NCT01835912.


Nano-Particles Carried by Multiple Dynein Motors Self-Regulate Their Number of Actively Participating Motors.
Intra-cellular active transport by native cargos is ubiquitous. We investigate the motion of spherical nano-particles (NPs) grafted with flexible polymers that end with a nuclear localization signal peptide. This peptide allows the recruitment of several mammalian dynein motors from cytoplasmic extracts. To determine how motor-motor interactions influenced motility on the single microtubule level, we conducted bead-motility assays incorporating surface adsorbed microtubules and combined them with model simulations that were based on the properties of a single dynein. The experimental and simulation results revealed long time trajectories: when the number of NP-ligated motors Nm increased, run-times and run-lengths were enhanced and mean velocities were somewhat decreased. Moreover, the dependence of the velocity on run-time followed a universal curve, regardless of the system composition. Model simulations also demonstrated left- and right-handed helical motion and revealed self-regulation of the number of microtubule-bound, actively transporting dynein motors. This number was stochastic along trajectories and was distributed mainly between one, two, and three motors, regardless of Nm. We propose that this self-regulation allows our synthetic NPs to achieve persistent motion that is associated with major helicity. Such a helical motion might affect obstacle bypassing, which can influence active transport efficiency when facing the crowded environment of the cell.


A model to calculate the induced dose rate around an 18 MV ELEKTA linear accelerator.
The dose rate due to activity induced by (gamma, n) reactions around an ELEKTA Precise accelerator running at 18 MV is reported. A model to calculate the induced dose rate for a variety of working practices has been derived and compared to the measured values. From this model, the dose received by the staff using the machine can be estimated. From measured dose rates at the face of the linear accelerator for a 10 x 10 cm2 jaw setting at 18 MV an activation coefficient per MU was derived for each of the major activation products. The relative dose rates at points around the linac head, for different energy and jaw settings, were measured. Dose rates adjacent to the patient support system and portal imager were also measured. A model to calculate the dose rate at these points was derived, and compared to those measured over a typical working week. The model was then used to estimate the maximum dose to therapists for the current working schedule on this machine. Calculated dose rates at the linac face agreed to within +/- 12% of those measured over a week, with a typical dose rate of 4.5 microSv h(-1) 2 min after the beam has stopped. The estimated maximum annual whole body dose for a treatment therapist, with the machine treating at only 18 MV, for 60000 MUs per week was 2.5 mSv. This compares well with value of 2.9 mSv published for a Clinac 21EX. A model has been derived to calculate the dose from the four dominant activation products of an ELEKTA Precise 18 MV linear accelerator. This model is a useful tool to calculate the induced dose rate around the treatment head. The model can be used to estimate the dose to the staff for typical working patterns.


[Panic attacks and panic disorder. Checklist for the diagnosis].
After looking for possible organic factors, other psychiatric disorders must be considered, e.g. depression or other anxiety disorders. With respect of the different neurobiologic and psychologic factors of panic attacks the therapy should be complex. Psychotherapy (e.g. behavior therapy) and psychopharmacologic treatments were proved very well. For the general practitioner the treatment with SSRI is very helpful. This approach provides a 60% full remission. For an uncomplicated disorder prophylactic treatment is necessary.


Comparing the biological impact of glatiramer acetate with the biological impact of a generic.
For decades, policies regarding generic medicines have sought to provide patients with economical access to safe and effective drugs, while encouraging the development of new therapies. This balance is becoming more challenging for physicians and regulators as biologics and non-biological complex drugs (NBCDs) such as glatiramer acetate demonstrate remarkable efficacy, because generics for these medicines are more difficult to assess. We sought to develop computational methods that use transcriptional profiles to compare branded medicines to generics, robustly characterizing differences in biological impact. We combined multiple computational methods to determine whether differentially expressed genes result from random variation, or point to consistent differences in biological impact of the generic compared to the branded medicine. We applied these methods to analyze gene expression data from mouse splenocytes exposed to either branded glatiramer acetate or a generic. The computational methods identified extensive evidence that branded glatiramer acetate has a more consistent biological impact across batches than the generic, and has a distinct impact on regulatory T cells and myeloid lineage cells. In summary, we developed a computational pipeline that integrates multiple methods to compare two medicines in an innovative way. This pipeline, and the specific findings distinguishing branded glatiramer acetate from a generic, can help physicians and regulators take appropriate steps to ensure safety and efficacy.


Triosephosphate isomerase (TPI) facilitates the replication of WSSV in Exopalaemon carinicauda.
Triosephosphate isomerase (TPI) is a vital enzyme in the glycolytic pathway, which can catalyze the interconversion of glyceraldehyde-3-phosphate (GAP) and dihydroxyacetone phosphate (DHAP). DHAP is involved in lipid metabolism and phospholipid synthesis. In order to know the role of TPI in WSSV infection to prawn, we cloned the full length cDNA of triosephosphate isomerase gene (EcTPI) from Exopalaemon carinicauda, and its function during WSSV infection was analyzed. EcTPI transcripts were widely distributed in all tissues, but showed relatively higher expression levels in the gill and epidermis. Its expression was apparently up-regulated after 24 h post WSSV injection (hpi), when the virus load began to rise. Furthermore, we detected the expressions of the key genes encoding the enzymes which catalyze the key steps in the glycolysis during WSSV infection. The data showed that genes encoding the enzymes which catalyzed upper steps of glycolysis to produce GAP, including hexokinase (HK), glucose-6-phosphate isomerase (GPI) and phosphofructokinase-1 (PFK-1), were significantly up-regulated at 24 and 27 hpi. Genes encoding the enzymes catalyzing down steps of glycolysis after GAP, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), enolase (ENO) and pyruvate kinase (PK), were apparent down-regulated at 24 and 27 hpi. Meanwhile, the gene encoding the enzyme glycerol-3-phosphate dehydrogenase (GPDH) catalyzing DHAP to glycerol-3-phosphate (G-3-P) showed down-regulation at 12-27 hpi, while the gene encoding dihydroxyacetone-phosphate acyltransferase (DHAPAT) catalyzing DHAP to further synthesis of phospholipids showed up-regulation at 12-24 hpi. These data suggested that WSSV infection could change the glycolysis pathway to make them produce more phospholipids which could be very helpful for virus replication. In order to further confirm the above speculation, dsRNA interference (RNAi) approach was used to knock down EcTPI gene and analyze its effect on WSSV load in prawn. The data showed that interference of EcTPI gene led to a significant decrease of WSSV loads in WSSV infected prawn. These data provided useful information to understand the infection mechanism of WSSV.


Deep learning reconstruction for improving the visualization of acute brain infarct on computed tomography.
This study aimed to investigate the impact of deep learning reconstruction (DLR) on acute infarct depiction compared with hybrid iterative reconstruction (Hybrid IR). This retrospective study included 29 (75.8 ± 13.2 years, 20 males) and 26 (64.4 ± 12.4 years, 18 males) patients with and without acute infarction, respectively. Unenhanced head CT images were reconstructed with DLR and Hybrid IR. In qualitative analyses, three readers evaluated the conspicuity of lesions based on five regions and image quality. A radiologist placed regions of interest on the lateral ventricle, putamen, and white matter in quantitative analyses, and the standard deviation of CT attenuation (i.e., quantitative image noise) was recorded. Conspicuity of acute infarct in DLR was superior to that in Hybrid IR, and a statistically significant difference was observed for two readers (p ≤ 0.038). Conspicuity of acute infarct with time from onset to CT imaging at < 24 h in DLR was significantly improved compared with Hybrid IR for all readers (p ≤ 0.020). Image noise in DLR was significantly reduced compared with Hybrid IR with both the qualitative and quantitative analyses (p < 0.001 for all). DLR in head CT helped improve acute infarct depiction, especially those with time from onset to CT imaging at < 24 h.


Recombinant T cell receptor molecules can prevent and reverse experimental autoimmune encephalomyelitis: dose effects and involvement of both CD4 and CD8 T cells.
Autoimmune diseases are often characterized by spontaneous remission followed by relapses. Although the mechanism(s) controlling pathogenic self-reactive T cells are not fully understood, recent data in experimental autoimmune encephalomyelitis (EAE), a prototype for CD4 T cell-mediated autoimmune diseases, indicate that spontaneous recovery is mediated by regulatory T cells (Treg) specific for peptides derived from the beta-chain of the TCR. Here we have tested whether recombinant single-chain TCRs (scTCRs) containing Vbeta domains can be used as vaccines for efficient priming of Treg. A single injection of mice with these recombinant proteins leads to efficient in vivo priming of Treg and almost complete protection from Ag-induced EAE. Significantly, administration of scTCRs during ongoing disease at a 10-fold lower dose than that required for prophylactic treatment also reverses established EAE. However, if a higher dose of scTCR is administered during ongoing disease, paralytic symptoms become exacerbated and the majority of treated animals die from severe and chronic EAE. Furthermore, we demonstrate that regulatory determinants are processed and presented from scTCRs resulting in the recruitment of both CD4 and CD8 regulatory T cells which are required for efficient regulation induced by scTCR. Reversal of established disease following an optimum dose of recombinant TCRs suggests that proteins expressing appropriate Vbeta domains could be used for the treatment of a variety of T cell-mediated pathologic conditions.


ZACD: A Retrograde Panoramic Analysis among Indian Population with New System of Classification.
Zygomatic Air Cell Defect (ZACD) has been considered as a normal radiographic variant with a spare knowledge. This should be judged before any surgical procedure of the temporal region as lack of which can lead to infections of cranium and may other complications. The present study was undertaken to estimate the prevalence and characteristics of ZACD and to propose a classification for the same using digital panoramic radiography. A total of 6825 digital panoramic radiographs were evaluated retrospectively of subjects aged between 4-90 years from both genders. The radiographs were evaluated by 4 (four) oral radiologists and the ZACD was recorded only when the decision was indisputable. Out of 6825 digital panoramic radiographs evaluated ZACD was noticed in 133 subjects with an overall prevalence of 1.94%. A male predilection was noted in the present study with the maximum prevalence during the second, third and fourth decade of life and as the age increases its prevalence decreases. Most commonly it was found to be unilateral and the unilocular presentation being more common. Most of the finding of the present study is in acceptance with those conducted previously, but some of the finding contradicts those in the previous ones. It can be because of low sample size of the previous studies. Moreover this study gives a new system to classify these defects to help diagnose the nature of the defect.


Autonomic responding to aversive words without conscious valence discrimination.
A growing body of data suggests that the emotional dimension of a stimulus can be processed without conscious identification of the stimulus. The arousal system could be activated by unrecognised biologically significant stimuli through simple physical stimulus features related to threat, without any evaluation of the meaning of the stimulus. However, unconscious processing of emotionally laden words cannot rely only on perceptual features but must include some analysis of symbolic meaning. The first aim of the present study was to assess whether masked (unrecognised) aversive words can elicit enhanced skin conductance responses (SCRs), a major autonomic index of emotional arousal, in normal participants. Our second aim was to determine whether any autonomic activation related to affective value of words is independent from access of this value to consciousness. Thus, the presentation duration of masked aversive and neutral words was determined, for each participant, in such a way that (1) identification was precluded, (2) valence discrimination was at chance, as indicated by performance in a forced-choice two-alternative task and by confidence ratings of the responses, and (3) emotional and neutral words were not detected differentially. SCRs were recorded during masked and unmasked presentations of both types of word. SCRs elicited by unmasked words, and also by masked words, were of greater magnitude when the words were emotional than when they were neutral. Consequently, in normal participants, autonomic activation can be a discriminative marker of the affective dimension of unrecognised verbal material in the absence of conscious valence identification.


The important role of radiation treatment in the management of Merkel cell carcinoma.
Merkel cell carcinoma is an aggressive, radiosensitive cutaneous neuroendocrine tumour. In this review, the roles of radiation therapy and chemoradiation in the management of Merkel cell carcinoma are described and discussed, and guidelines for patient management are presented. Radiation treatment may be indicated for definitive (> 55 Gy) or adjuvant (> 50 Gy) treatment of the primary tumour site and for prophylactic (> 50 Gy), adjuvant (> 50 Gy) or definitive (> 55 Gy) treatment of the regional lymph node field. If a patient presents with positive margins after initial biopsy or resection, definitive radiation therapy or chemoradiation may be an alternative to further surgery and, importantly, results in less delay than re-resection followed by adjuvant radiation treatment. Given the rarity of this tumour, patients should be enrolled on prospective databases and clinical trials, and managed in a multidisciplinary clinical setting wherever possible.


Leishmania promastigotes induce cytokine secretion in macrophages through the degradation of synaptotagmin XI.
Synaptotagmins (Syts) are type-I membrane proteins that regulate vesicle docking and fusion in processes such as exocytosis and phagocytosis. We recently discovered that Syt XI is a recycling endosome- and lysosome-associated protein that negatively regulates the secretion of TNF and IL-6. In this study, we show that Syt XI is directly degraded by the zinc metalloprotease GP63 and excluded from Leishmania parasitophorous vacuoles by the promastigotes surface glycolipid lipophosphoglycan. Infected macrophages were found to release TNF and IL-6 in a GP63-dependent manner. To demonstrate that cytokine release was dependent on GP63-mediated degradation of Syt XI, small interfering RNA-mediated knockdown of Syt XI before infection revealed that the effects of small interfering RNA knockdown and GP63 degradation were not cumulative. In mice, i.p. injection of GP63-expressing parasites led to an increase in TNF and IL-6 secretion and to an augmented influx of neutrophils and inflammatory monocytes to the inoculation site. Both of these cell types have been shown to be infection targets and aid in the establishment of infection. In sum, our data revealed that GP63 induces proinflammatory cytokine release and increases infiltration of inflammatory phagocytes. This study provides new insight on how Leishmania exploits the immune response to establish infection.


Angles no longer weigh in: The effect of geometric cue directness on reorientation.
Previous research in spatial reorientation, which only presented the target location in the corner, has found that adults weighed angles more than wall lengths. We proposed that in previous research, angular cues were available for direct use whereas length cues had to be associated with the left/right sense. We thus investigated whether the directness of cues rather than the cues themselves accounted for the previous findings in the reorientation task. Through navigating a virtual environment, 111 participants were trained to remember target locations in a parallelogram-shaped room and tested in varied versions of the training environment: (a) a reverse-parallelogram environment where angular information conflicted with wall length information, (b) a rhombic environment that preserved only angular information from the training environment, and (c) a rectangular environment that preserved only wall length information. We varied the directness of the two cues in the current study. In addition to the condition with target location in the corner, we included a condition that placed the target positions in the middle of the walls, making the length cues direct. The results revealed that angular information no longer received priority, especially in the wall condition. More interestingly, compared to the group trained with the target positions at walls, the group trained with target positions at corners weighted angular cues more heavily in conflict trials and performed less accurately using length cues in rectangular environments. The results suggest that human adults prefer to rely on direct cues more than indirect cues in the reorientation process. (PsycINFO Database Record


Effects of Oxalobacter formigenes in subjects with primary hyperoxaluria Type 1 and end-stage renal disease: a Phase II study.
In primary hyperoxaluria Type 1 (PH1), endogenous oxalate overproduction significantly elevates urinary oxalate excretion, resulting in recurrent urolithiasis and/or progressive nephrocalcinosis and often early end-stage renal disease (ESRD). In ESRD, dialysis cannot sufficiently remove oxalate; plasma oxalate (Pox) increases markedly, inducing systemic oxalate deposition (oxalosis) and often death. Interventions to reduce Pox in PH1 subjects with ESRD could have significant clinical impact. This ongoing Phase II, open-label trial aimed to evaluate whether long-term Oxabact™ (Oxalobacter formigenes, OC5, OxThera Intellectual Property AB, Sweden) lowers Pox in PH1 ESRD subjects, ameliorating clinical outcome. PH1 ESRD subjects on stable dialysis regimens were examined. Subjects were administered one OC5 capsule twice daily for up to 36 months or until transplantation. Total Pox values, cardiac function and safety were evaluated. Free Pox was evaluated in a comparative non-treated PH1 dialysis group using retrospective chart reviews and analyses. Twelve subjects enrolled in an initial 6-week treatment phase. Following a washout of up to 4 weeks, eight subjects entered a continuation study; outcomes after 24 months of treatment are presented. After 24 months, all subjects had reduced or non-elevated Pox compared with baseline. Cardiac function improved, then stabilized. No treatment-related serious adverse events were reported. Compared with an untreated natural control cohort, 24 months OC5 administration was beneficial to PH1 ESRD subjects by substantially decreasing Pox concentrations, and improving or stabilizing cardiac function and clinical status, without increasing dialysis frequency. OC5 was safe and well-tolerated.


Sustainable approach to oil recovery from oil spills through superhydrophobic jute fabric.
Ecosystems suffer from increased oil exploitation and frequent oil spills, which calls for effective, environment-friendly, and economically viable solutions. To address this, abandoned gunny sacks as the concerned jute fabric were superhydrophobically (water contact angle ∼159°) modified, incorporating titanium dioxide (TiO2) nanoparticles and hexadecyltrimethoxysilane (HDTMS), rendering a facile drop casting procedure. The modified superhydrophobic-superoleophilic jute fabric has been identified as a high-performance filter with superior reusability that can separate oil-water mixtures in challenging environmental conditions (including potent acidic, alkaline, highly saline, aqueous, frigid, and blistering water environments) while maintaining high separation efficiency. In continuation, static conditions indulging a batch and continuous oil separation performance and dynamic conditions stimulating turbulence in the oil-water mixture were proficiently carried out, mimicking real-world circumstances. As a result, the modified jute fabric has the advantages of high separation efficiency, stable recyclable properties, and outstanding durability, highlighting its enormous potential for use in practical applications.


Hierarchically designed electrospun tubular scaffolds for cardiovascular applications.
Hierarchically designed tubular scaffolds with bi-layer and multi-layer structures are expected to mimic native vessels in its structural geometry. A new approach for the fabrication of hierarchically designed tubular scaffold with suitable morphology was introduced through electrospinning technique. Among these scaffolds, bi-layer scaffold had a single inner and outer layer whereas multilayer scaffold had more number of inner layers. The inner layer/layers of the scaffolds were made up of aligned poly (lactic acid) (PLA) fibers for EC adhesion where as outer layers were composed of random fibers of poly (caprolactone) (PCL) and PLA providing larger pores for SMC penetration. The fabricated scaffolds were characterized by FTIR spectroscopy and Differential Thermal Analysis (DTA) and examined by evaluating cellular interactions. Human Umbilical Vein Endothelial Cells (HUVECs) seeded on aligned PLA fibers showed enhanced cellular orientation and cytoskeletal organization. In addition, the PCL-PLA composite random fibers supported SMC adhesion and proliferation sufficiently. The functionality of the endothelial cells grown on the PLA-aligned scaffold was also found to be satisfactory. Lining the constructs with a luminal monolayer of well-organized ECs along with homogenously distributed SMCs surrounding them might result in vascular conduits suitable for in vivo applications. Since this hierarchically designed tubular scaffold closely mimics the morphology of native vessel, this could be a better candidate for vascular tissue engineering.


The interactive effects of exercise type and environment temperature on HSP72 in active females.
The purpose of this study was to determine the effects of endurance and/ or weight training in normal and/ or moderate heat environments on heat shock protein (HSP72) response in active young females. Forty-five healthy active female (21±3 year, 61.4±10.2 kg, VO2max 37.2±5.3 mL/kg/min) were randomLy assigned into five groups; endurance training+normal heat (ET+NH), endurance training+moderate heat (ET+MH), weight training+normal heat (WT+NH), weight training+moderate heat (WT+MH) and heat group (HG). The training protocols include treadmill running to exhaustion on a treadmill at %65-75 V02max in the ET+NH and ET+MH groups and 4 sets of eccentric actions of elbow flexors (two sets with %50 and two sets with %60 1RM) in the WT+NH and WT+MH groups. Fasting blood samples were collected from non-dominant arm at three phases; consist of rest (baseline) and mid-test and 30 min after the training protocols. Endurance training and/ or weight training, in particularly, in the moderate heat ambient resulted in an increase in the HSP72, creatin kinase (CK) and cortisol values, as compared to ET+NH and WT+NH groups and the mid-test and baseline phases. However, the HSP72 and CK concentrations significantly increased in WT+ NH group, as compared to ET+NH group. Furthermore, WT+MH group showed significant increase in HSP72 levels, as compared to ET+ NH group. These findings suggest the muscle damage related to factors such as; exercise type, environment temperature and increase in exercise-induced body temperature.


Mycobacterium tuberculosis-Induced Polarization of Human Macrophage Orchestrates the Formation and Development of Tuberculous Granulomas In Vitro.
The tuberculous granuloma is an elaborately organized structure and one of the main histological hallmarks of tuberculosis. Macrophages, which are important immunologic effector and antigen-presenting cells, are the main cell type found in the tuberculous granuloma and have high plasticity. Macrophage polarization during bacterial infection has been elucidated in numerous recent studies; however, macrophage polarization during tuberculous granuloma formation and development has rarely been reported. It remains to be clarified whether differences in the activation status of macrophages affect granuloma formation. In this study, the variation in macrophage polarization during the formation and development of tuberculous granulomas was investigated in both sections of lung tissues from tuberculosis patients and an in vitro tuberculous granuloma model. The roles of macrophage polarization in this process were also investigated. Mycobacterium tuberculosis (M. tuberculosis) infection was found to induce monocyte-derived macrophage polarization. In the in vitro tuberculous granuloma model, macrophage transformation from M1 to M2 was observed over time following M. tuberculosis infection. M2 macrophages were found to predominate in both necrotic and non-necrotic granulomas from tuberculosis patients, while both M1 and M2 polarized macrophages were found in the non-granulomatous lung tissues. Furthermore, it was found that M1 macrophages promote granuloma formation and macrophage bactericidal activity in vitro, while M2 macrophages inhibit these effects. The findings of this study provide insights into the mechanism by which M. tuberculosis circumvents the host immune system as well as a theoretical foundation for the development of novel tuberculosis therapies based on reprogramming macrophage polarization.


VEGF and soluble VEGF receptor-1 (sFlt-1) distributions in peripheral arterial disease: an in silico model.
Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis, the growth of new capillaries from existing microvasculature. In peripheral arterial disease (PAD), lower extremity muscle ischemia develops downstream of atherosclerotic obstruction. A working hypothesis proposed that the maladaptive overexpression of soluble VEGF receptor 1 (sVEGFR1) in ischemic muscle tissues, and its subsequent antagonism of VEGF bioactivity, may contribute to the deficient angiogenic response in PAD, as well as the limited success of therapeutic angiogenesis strategies where exogenous VEGF genes/proteins are delivered. The objectives of this study were to develop a computational framework for simulating the systemic distributions of VEGF and sVEGFR1 (e.g., intramuscular vs. circulating, free vs. complexed) as observed in human PAD patients and to serve as a platform for the systematic optimization of diagnostic tools and therapeutic strategies. A three-compartment model was constructed, dividing the human body into the ischemic calf muscle, blood, and the rest of the body, connected through macromolecular biotransport processes. Detailed molecular interactions between VEGF, sVEGFR1, endothelial surface receptors (VEGFR1, VEGFR2, NRP1), and interstitial matrix sites were modeled. Our simulation results did not support a simultaneous decrease in plasma sVEGFR1 during PAD-associated elevations in plasma VEGF reported in literature. Furthermore, despite the overexpression in sVEGFR1, our PAD control demonstrated increased proangiogenic signaling complex formation, relative to our previous healthy control, due to sizeable upregulations in VEGFR2 and VEGF expression, thus leaving open the possibility that impaired angiogenesis in PAD may be rooted in signaling pathway disruptions downstream of ligand-receptor binding.


Changes in peritoneal dialysis practices in Canada 1996-1999.
Over the past decade, clinical studies and clinical practice guidelines have suggested the use of higher small solute clearance targets for patients on peritoneal dialysis (PD). This study asks whether these recommendations have translated into changes in clinical prescription of PD. Data were collected annually from 1996 to 1999 on all prevalent dialysis patients in 24 Canadian centers, accounting for approximately 40% of the Canadian chronic dialysis population. Approximately a third of these patients were on PD. Full details of each patient's prescription were recorded, with particular attention to dwell volumes and frequency of exchanges for continuous ambulatory PD (CAPD) and to total treatment volumes and day dwells for automated PD (APD). The most recent Kt/V and creatinine clearance values available were recorded for each patient and the overall results for each year were compared to present treatment recommendations. 24 university- and community-based hospitals. From 1996 to 1999, the use of APD, relative to CAPD, grew from 14% to 28% of all PD patients. Among CAPD patients, the proportion using dwell volumes greater than 2 L rose from 14% to 32%, and the proportion doing more than 4 dwells per day rose from 16% to 28%. The mean daily volume of prescribed fluid for CAPD patients increased from 8.3 to 9.1 L. As a result, the proportion of patients achieving a weekly Kt/V above 2.0 rose from 54% to 72%, and those receiving a Kt/V less than 1.7 fell from 22% to 10%. For creatinine clearance, those exceeding 60 L per week rose from 63% to 73%. For APD, the mean treatment volume rose from 11.8 L in 1996 to plateau at about 13.4 L in 1998 and 1999. However, the proportion of patients receiving more than 1 day dwell grew from 31% in 1998 to 40% in 1999, and the proportion that were "day dry" fell from 25% to 17%. For APD, the proportion of patients with a Kt/V above 2.0 rose from 67% to 77%, and with a creatinine clearance above 60 L, from 62% to 70%. The proportion with no recent clearance value recorded fell during the course of the study, from 45% to 27%. There was a marked change in PD prescription practices in Canada during the second half of the 1990s. This occurred in response to clinical studies and publication of guidelines. There is room for further improvement, especially with respect to the proportion of patients that did not have regular clearance measurements made.


Morphological and functional aspects of progenitors perturbed in cortical malformations.
In this review, we discuss molecular and cellular mechanisms important for the function of neuronal progenitors during development, revealed by their perturbation in different cortical malformations. We focus on a class of neuronal progenitors, radial glial cells (RGCs), which are renowned for their unique morphological and behavioral characteristics, constituting a key element during the development of the mammalian cerebral cortex. We describe how the particular morphology of these cells is related to their roles in the orchestration of cortical development and their influence on other progenitor types and post-mitotic neurons. Important for disease mechanisms, we overview what is currently known about RGC cellular components, cytoskeletal mechanisms, signaling pathways and cell cycle characteristics, focusing on how defects lead to abnormal development and cortical malformation phenotypes. The multiple recent entry points from human genetics and animal models are contributing to our understanding of this important cell type. Combining data from phenotypes in the mouse reveals molecules which potentially act in common pathways. Going beyond this, we discuss future directions that may provide new data in this expanding area.


Markers of bone metabolism are affected by renal function and growth hormone therapy in children with chronic kidney disease.
The extent and relevance of altered bone metabolism for statural growth in children with chronic kidney disease is controversial. We analyzed the impact of renal dysfunction and recombinant growth hormone therapy on a panel of serum markers of bone metabolism in a large pediatric chronic kidney disease cohort. Bone alkaline phosphatase (BAP), tartrate-resistant acid phosphatase 5b (TRAP5b), sclerostin and C-terminal FGF-23 (cFGF23) normalized for age and sex were analyzed in 556 children aged 6-18 years with an estimated glomerular filtration rate (eGFR) of 10-60 ml/min/1.73 m2. 41 children receiving recombinant growth hormone therapy were compared to an untreated matched control group. Standardized levels of BAP, TRAP5b and cFGF-23 were increased whereas sclerostin was reduced. BAP was correlated positively and cFGF-23 inversely with eGFR. Intact serum parathormone was an independent positive predictor of BAP and TRAP5b and negatively associated with sclerostin. BAP and TRAP5B were negatively affected by increased C-reactive protein levels. In children receiving recombinant growth hormone, BAP was higher and TRAP5b lower than in untreated controls. Sclerostin levels were in the normal range and higher than in untreated controls. Serum sclerostin and cFGF-23 independently predicted height standard deviation score, and BAP and TRAP5b the prospective change in height standard deviation score. Markers of bone metabolism indicate a high-bone turnover state in children with chronic kidney disease. Growth hormone induces an osteoanabolic pattern and normalizes osteocyte activity. The osteocyte markers cFGF23 and sclerostin are associated with standardized height, and the markers of bone turnover predict height velocity.


Adeno-associated virus type 2-mediated transfer of ecotropic retrovirus receptor cDNA allows ecotropic retroviral transduction of established and primary human cells.
The cellular receptors that mediate binding and internalization of retroviruses have recently been identified. The concentration and accessibility of these receptors are critical determinants in accomplishing successful gene transfer with retrovirus-based vectors. Murine retroviruses containing ecotropic glycoproteins do not infect human cells since human cells do not express the receptor that binds the ecotropic glycoproteins. To enable human cells to become permissive for ecotropic retrovirus-mediated gene transfer, we have developed a recombinant adeno-associated virus type 2 (AAV) vector containing ecotropic retroviral receptor (ecoR) cDNA under the control of the Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter (vRSVp-ecoR). Established human cell lines, such as HeLa and KB, known to be nonpermissive for murine ecotropic retroviruses, became permissive for infection by a retroviral vector containing a bacterial gene for resistance to neomycin (RV-Neo(r)), with a transduction efficiency of up to 47%, following transduction with vRSVp-ecoR, as determined by the development of colonies that were resistant to the drug G418, a neomycin analog. No G418-resistant colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. Southern and Northern blot analyses revealed stable integration and long-term expression, respectively, of the transduced murine ecoR gene in clonal isolates of HeLa and KB cells. Similarly, ecotropic retrovirus-mediated Neo(r) transduction of primary human CD34+ hematopoietic progenitor cells from normal bone marrow was also documented, but only following infection with vRSVp-ecoR. The retroviral transduction efficiency was approximately 7% without prestimulation and approximately 14% with prestimulation of CD34+ cells with cytokines, as determined by hematopoietic clonogenic assays. No G418-resistant progenitor cell colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. These results suggest that sequential transduction of primary human cells with two different viral vectors may overcome limitations encountered with a single vector. Thus, the combined use of AAV- and retrovirus-based vectors may have important clinical implications for ex vivo and in vivo human gene therapy.