Impact of highly active antiretroviral therapy on organ-specific manifestations of HIV-1 infection.
In the last 10 years, interesting results have been reported concerning the impact of highly active antiretroviral therapy (HAART) on the changing pattern of organ-specific manifestations of HIV-1 infection. There has been a clear step-wise reduction in the incidence of several opportunistic infections (OIs), particularly Pneumocystis carinii pneumonia, whereas a nonsignificant reduction in incidence has been observed for other organ-specific diseases, including invasive cervical cancer and Hodgkin disease. In addition, several organ-specific manifestations, including HIV-associated nephropathy, wasting syndrome and cardiomyopathy, are a direct consequence of damage by HIV-1, and so HAART may have a therapeutic effect in improving or preventing these manifestations. Finally, the introduction of HAART has seen the emergence of several complications, termed immune reconstitution inflammatory syndrome, which includes OIs such as cytomegalovirus vitritis, Mycobacterium avium complex lymphadenitis, paradoxical responses to treatment for tuberculosis, and exacerbation of cryptococcosis. Because not all HIV-1 organ-specific manifestations are decreasing in the HAART era, this review will analyse the influence of HAART on several organ-specific manifestations, and in particular OIs related to several organs, cerebral disorders and HIV-1-related neoplasia.


[Reticuloendotheliosis virus infection in ducks--an epidemiological studies].
To study the epidemiological status of reticuloendotheliosis virus (REV) infection in ducks. Two hundred and twenty tissue samples of the Bursa, spleens and livers were randomly collected from 97 sick or dead ducks from 3 areas in Shandong Province, China. REV infections were tested by virus isolation in cell cultures, direct dot blot hybridization, regular PCR and nest-PCR with tissue-extracted DNA as the templates. PCR products of env gene fragments were cloned and sequenced, sequences of env gene fragments were compared and analyzed. REV infections were detected in 35/39 (90%) bursal samples, 54/84 (64%) spleens and 32/97 (33%) livers by Nest-PCR. The positive rate of bursal samples was significantly higher than spleen and liver samples (P < 0.01). No positive sample was detected by virus isolation, direct dot blot hybridization or regular PCR from the same Bursa, spleen and liver samples. In sequence comparisons of amplified env gene fragments, Yinan-1 and Binzhou-1 had 99.8% of identity with reference SNV strain isolated from ducks in USA, and Linqu-1 had 100% identity with two other reference strains isolated from chickens in USA. However, they had lower homology with strains isolated from chickens in China. More attention should be given to bursal samples in detection of REV. Since REV was existing only at a very low level in infected samples and difficult to be detected, nest-PCR should be conducted as the more sensitive assay. Sequence homology comparisons and phylogenetic tree analysis suggested that REV infections in these ducks may originally come from imported breeder ducklings without inspection for REV.


Effect of image magnification of digitized bitewing radiographs on approximal caries detection: an in vitro study.
To evaluate whether or not image magnification of digitized bitewing radiographs had an effect on approximal caries detection. Twenty general dental practitioners (GDPs) rated a total of 140 digital images (28 approximal surfaces at five different magnifications, x3, x6, x12, x18 and x30) for caries on a 5-point confidence scale. The observations were compared with the state of the approximal surfaces on visual inspection. For each GDP the average response rate was calculated. Paired t-tests were used to compare the diagnostic response at the five different image magnifications. The mean response rates obtained at x18 and x30 magnifications were significantly inferior to those at the three smaller magnifications (p < 0.007). Digital image magnification has a significant influence on observer performance in the detection of approximal caries, with an upper limit beyond which diagnostic accuracy may be reduced. Further research is needed to determine limits for various different diagnostic tasks so as to give GDPs guidelines on the appropriate magnifications to use.


Genomic Epidemiology of Azithromycin-Nonsusceptible Neisseria gonorrhoeae, Argentina, 2005-2019.
Azithromycin-nonsusceptible Neisseria gonorrhoeae strains are an emerging global public health threat. During 2015-2018, the prevalence of azithromycin-nonsusceptible gonococcal infection increased significantly in Argentina. To investigate the genomic epidemiology and resistance mechanisms of these strains, we sequenced 96 nonsusceptible isolates collected in Argentina during 2005-2019. Phylogenomic analysis revealed 2 main clades, which were characterized by a limited geographic distribution, circulating during January 2015-November 2019. These clades included the internationally spreading multilocus sequence types (STs) 1580 and 9363. The ST1580 isolates, which had MICs of 2-4 μg/mL, had mutations in the 23S rRNA. The ST9363 isolates, which had MICs of 2-4 or >256 μg/mL, had mutations in the 23S rRNA, a mosaic mtr locus, or both. Identifying the geographic dissemination and characteristics of these predominant clones will guide public health policies to control the spread of azithromycin-nonsusceptible N. gonorrhoeae in Argentina.


Endovascular aneurysm repair: Magnetic resonance monitoring of histological organization processes in the excluded aneurysm.
The purpose of the present study was to systematically analyze the histopathologic organization processes in excluded aneurysms after endovascular stenting and to develop a noninvasive monitoring method for these processes using MRI. In 36 mongrel dogs, autologous aortic aneurysms were created. Endovascular treatment was performed using covered stents. Follow-up was after 1 week, 6 weeks, and 6 months. MRI was performed with T2-weighted turbo-spin-echo sequences and T1-weighted spin-echo sequences and was repeated after contrast bolus with gadolinium. Histopathologic findings were correlated to signal intensities (SIs) of MRI images. SIs of distinct areas were analyzed and related to the SI of the reference tissue (SI ratio). The histological organization process was gradated in the following 4 classes: class 0, detritus without organization; classes I and II, connective tissue proliferation with increasing fiber synthesis; and class III, dense fibrous connective tissue. The SI ratios of T2-weighted images were significantly reduced from 4.76 in detritus (0) to 1.70 in dense fibrous connective tissue (III) as a function of histopathologic classes. SI ratios of T1-weighted images were reduced from 1.84 (0) to 1.12 (III). Contrast bolus with gadolinium-DTPA showed no change of SI ratio in detritus (0.99) but an increase from 1.12 (I) to 1.70 (III) as organization increased. The histological organization of excluded aneurysms can be monitored by MRI. Progressive organization is indicated by decreasing SIs in T2- and an increasing signal increase in T1-weighted images after gadolinium bolus.


Virilization in a Postmenopausal Female Due to Androgen Secreting Ovarian Dermoid Cyst.
Virilizing ovarian dermoid cysts are very rare. The source of androgen in these cysts may be tumors such as Sertoli-Leydig cell tumor or Leydig cell hyperplasia. A 52-year-old postmenopausal female with virilization was found to have an ovarian dermoid cyst on ultrasound. Her serum testosterone levels were elevated. Leydig cell hyperplasia within the dermoid cyst was found to be the source of androgen in this patient.


Insight Into Neonatal Sepsis: An Overview.
There are approximately 1.3 million cases of neonatal sepsis reported worldwide with deaths occurring more commonly in preterm and low-weight newborns. Neonatal sepsis is the third major cause of neonatal deaths resulting in 203,000 deaths per year. It is divided into two subtypes based on time of occurrence: early-onset neonatal sepsis (ENS), occurring within the first 72 hours of birth usually due to perinatal risk factors, and late-onset neonatal sepsis (LOS) usually occurring after the first week of life and up to 28th day of life. There are many complications associated with neonatal sepsis including septic shock, multiple organ failure, and death. It is vital for clinicians to know the signs and symptoms of neonatal sepsis in order to diagnose it early. Preventive measures, early diagnosis, appropriate antibiotic administration, timely supportive management, and the establishment of efficient management are vital in the prevention of severe complications or death. In this review, we aim to provide the most up-to-date information regarding risk factors, pathophysiology, signs and symptoms, diagnosis, and treatment of neonatal sepsis. We discuss the maternal and neonatal risk factors involved in the pathogenesis of neonatal sepsis and the signs and symptoms of early and late neonatal sepsis. We focus on the different pathogens involved and the markers used in the diagnosis and treatments available for each.


Stress and satisfaction among employees in AIDS service organizations in New York.
In recent years, employees in AIDS service organizations have witnessed a number of changes and challenges in their work, ranging from shifts in the population primarily affected by HIV disease, treatment advances and its implications, and changes in administrative factors such as cuts in funding, reorganization, and changes in job function. This study examined the motives, stressors, and rewards of employees in AIDS service organizations in New York City. The main reasons for choosing to work in this field were a desire to help and having experienced the loss of some-one to AIDS. A large number of respondents had been working in the AIDS field for 5 or more years and perceived the level of stress to be mild to moderate. They reported that the rewards of this type of work outweighed the stressors, and the level of job satisfaction was very high.


The Na+/Ca2+ Exchanger 1 (NCX1) Variant 3 as the Major Extrusion System in Renal Distal Tubular Transcellular Ca2+-Transport.
Fine-tuning of renal calcium (Ca(2+)) reabsorption takes place in the late distal convoluted and connecting tubules (DCT2/CNT) of the kidney via transcellular Ca(2+) transport. Here, Ca(2+) enters the cell at the apical side via the epithelial Ca(2+) channel transient receptor potential vanilloid 5 and is subsequently extruded at the basolateral side by the concerted actions of the plasma membrane Ca(2+) ATPases and the Na(+)/Ca(2+) exchanger 1 (NCX1). NCX1 is responsible for ∼ 70% of basolateral Ca(2+) extrusion. The aim of this study was to determine the predominant NCX1 variant in the kidney and its role in Ca(2+) transport. DCT2/CNT specific tubules were used to show the abundance of NCX1 specific isoforms. Renal NCX1 variants were cloned from mouse kidney tissue. Human Embryonic Kidney 293(T) cells were transiently transfected with NCX1.3, and Fura-2 measurements and 45Ca(2+) uptake assays were performed to determine several characteristics of NCX1.3 in the reverse mode. NCX1.3 was demonstrated to be the predominant NCX1 variant in the DCT2/CNT, next to NCX1.2 and NCX1.7. NCX1.3 could be inhibited by SN-6, an NCX-specific inhibitor, whereas stimulation of the cAMP/PKA or PKC-mediated pathway did not affect Ca(2+) influx as measured in the reverse mode. Lowering intracellular Ca(2+) concentrations resulted in a decreased Ca(2+) uptake. NCX1.3 is the predominant NCX variant in the DCT2/CNT tubules. Its function is dependent on intracellular Ca(2+) concentrations.


On-line computer scoring of the auditory brainstem response for estimation of hearing threshold.
Auditory brainstem responses (ABRs) to click stimuli have become established as a useful indicator of hearing thresholds in infants and young children. Although the investigation is objective in so far as the patient is concerned, a subjective element remains in the decision by the operator as to whether or not an ABR is present in an averaged waveform. A simple on-line computer detection technique is described which removes some of the reliance placed upon the operator. The technique employs a scanning window for correlation and amplitude analysis of pairs of averaged ABR waveforms. The reliability and accuracy of computer and operator scoring of 25 thresholds in normally hearing adults and 50 thresholds in infants and young children with suspected hearing impairment have been investigated. In the adult group, 96% of computer estimates of the threshold were within +/- 10 dB of the subjective hearing threshold, compared with 92% for operator scoring. There were 92% of computer and operator scores within +/- 10 dB of each other. In the patient group there was equally good agreement between computer and operator scoring with 90% of the thresholds within +/- 10 dB of each other; the incidence of possible false-positive thresholds was also lower with computer scoring. This on-line scoring technique, therefore, offers useful assistance to the operator; requires only limited computing power, and is suitable for use in a routine clinical environment.


Sulphhydryl and disulphide stainings of amyloid: a comparison with hyaline body.
Histochemical staining for sulphhydryl (-SH) groups and disulphide (S-S) bonds was undertaken in 10 cases of cutaneous amyloidosis using the new fluorescent thiol reagent. A weak S-S fluorescence (but not -SH) was detected in dermal amyloid, suggesting that cross-linking of -SH groups may occur in amyloidogenesis. Since in normal skin S-S bonds were stained only in the horny layer and negligibly in the dermis, our findings seem to support the view that amyloid is derived at least partially from epidermal cells.


Testicular toxicity following separate and combined administration of PDE5 inhibitors and opioid: assessment of recovery following their withdrawal.
We previously observed that PDE5 inhibitors and opioids were widely abused in Nigeria. Here, we examined the effect of high doses of sildenafil, tadalafil, tramadol and sildenafil + tramadol on reproductive toxicity in male rats. Rats were either administered normal saline (0.2 ml), sildenafil (10 mg/kg), tadalafil (10 mg/kg), tramadol (20 mg/kg) or sildenafil + tramadol (10 and 20 mg/kg respectively) p. o. for 8 weeks. The recovery groups were allowed 8-week recovery period before sacrifice. Results showed that body weight change, testicular and epididymal weights, epididymal sperm count and sperm viability were significantly reduced in all treated groups compared with the control. Spermatozoa with abnormal morphology were significantly increased in all treated groups compared with the control. Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) were significantly reduced, while malondialdehyde (MDA) was significantly increased in all treated groups compared with the control. The severity of toxicity was highest in sildenafil + tramadol group. There was no complete recovery from reproductive toxicity following withdrawal of the various treatments. High doses of sildenafil, tadalafil, tramadol or sildenafil + tramadol result in testicular oxidative stress-induced reproductive toxicity with poor reversal following withdrawal.


Age affects the perception of limb length discrepancy in patients with and without a total hip arthroplasty.
The amount of lengthening or shortening that can be detected by patients before and after total hip arthroplasty has not been yet quantified. We studied the ability to detect limb length inequality in 194 patients with and without a total hip arthroplasty, match-paired for age and sex. None of the participants had clinical signs of lumbosacral pathology, spinal deformity, or fixed pelvic obliquity; and all had equal functional and actual limb length. The participants walked with shoes, with and without the addition of fixed insoles, to simulate 2.5, 5, 10 and 15 millimeters of shortening and lengthening of the tested limb. Lengthening and shortening were similarly detected. Younger individuals detected the differences better than older ones (p=0.001), and there was a significant correlation between the decade of life and the ability to detect a limb length discrepancy (r=-0.22; p=0.002). This study demonstrates that perception of limb length is affected by the age, with older individuals having less awareness of changes in limb length than younger ones. (Hip International 2004; 14: 249-53).


Identification and characterization of genes encoding the nuclear envelope LINC complex in the monocot species Zea mays.
The linker of nucleoskeleton to cytoskeleton (LINC) complex is an essential multi-protein structure spanning the nuclear envelope. It connects the cytoplasm to the nucleoplasm, functions to maintain nuclear shape and architecture and regulates chromosome dynamics during cell division. Knowledge of LINC complex composition and function in the plant kingdom is primarily limited to Arabidopsis, but critically missing from the evolutionarily distant monocots, which include grasses, the most important agronomic crops worldwide. To fill this knowledge gap, we identified and characterized 22 maize genes, including a new grass-specific KASH gene family. By using bioinformatic, biochemical and cell biological approaches, we provide evidence that representative KASH candidates localize to the nuclear periphery and interact with Zea mays (Zm)SUN2 in vivo FRAP experiments using domain deletion constructs verified that this SUN-KASH interaction was dependent on the SUN but not the coiled-coil domain of ZmSUN2. A summary working model is proposed for the entire maize LINC complex encoded by conserved and divergent gene families. These findings expand our knowledge of the plant nuclear envelope in a model grass species, with implications for both basic and applied cellular research.This article has an associated First Person interview with the first author of the paper.


Direct electrochemistry of hemoglobin immobilized on a functionalized multi-walled carbon nanotubes and gold nanoparticles nanocomplex-modified glassy carbon electrode.
Direct electron transfer of hemoglobin (Hb) was realized by immobilizing Hb on a carboxyl functionalized multi-walled carbon nanotubes (FMWCNTs) and gold nanoparticles (AuNPs) nanocomplex-modified glassy carbon electrode. The ultraviolet-visible absorption spectrometry (UV-Vis), transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) methods were utilized for additional characterization of the AuNPs and FMWCNTs. The cyclic voltammogram of the modified electrode has a pair of well-defined quasi-reversible redox peaks with a formal potential of -0.270 ± 0.002 V (vs. Ag/AgCl) at a scan rate of 0.05 V/s. The heterogeneous electron transfer constant (ks) was evaluated to be 4.0 ± 0.2 s(-1). The average surface concentration of electro-active Hb on the surface of the modified glassy carbon electrode was calculated to be 6.8 ± 0.3 × 10(-10) mol cm(-2). The cathodic peak current of the modified electrode increased linearly with increasing concentration of hydrogen peroxide (from 0.05 nM to 1 nM) with a detection limit of 0.05 ± 0.01 nM. The apparent Michaelis-Menten constant (K(m)(app)) was calculated to be 0.85 ± 0.1 nM. Thus, the modified electrode could be applied as a third generation biosensor with high sensitivity, long-term stability and low detection limit.


Grueneisen relaxation photoacoustic microscopy in vivo.
Grueneisen relaxation photoacoustic microscopy (GR-PAM) can achieve optically defined axial resolution, but it has been limited to ex vivo demonstrations so far. Here, we present the first in vivo image of a mouse brain acquired with GR-PAM. To induce the GR effect, an intensity-modulated continuous-wave laser was employed to heat absorbing objects. In phantom experiments, an axial resolution of 12.5 μm was achieved, which is sixfold better than the value achieved by conventional optical-resolution PAM. This axial-resolution improvement was further demonstrated by imaging a mouse brain in vivo, where significantly narrower axial profiles of blood vessels were observed. The in vivo demonstration of GR-PAM shows the potential of this modality for label-free and high-resolution anatomical and functional imaging of biological tissues.


Molecular mechanisms of alcoholic fatty liver.
Alcoholic fatty liver is a potentially pathologic condition which can progress to steatohepatitis, fibrosis, and cirrhosis if alcohol consumption is continued. Alcohol exposure may induce fatty liver by increasing NADH/NAD(+) ratio, increasing sterol regulatory element-binding protein-1 (SREBP-1) activity, decreasing peroxisome proliferator-activated receptor-alpha (PPAR-alpha) activity, and increasing complement C3 hepatic levels. Alcohol may increase SREBP-1 activity by decreasing the activities of AMP-activated protein kinase and sirtuin-1. Tumor necrosis factor-alpha (TNF-alpha) produced in response to alcohol exposure may cause fatty liver by up-regulating SREBP-1 activity, whereas betaine and pioglitazone may attenuate fatty liver by down-regulating SREBP-1 activity. PPAR-alpha agonists have potentials to attenuate alcoholic fatty liver. Adiponectin and interleukin-6 may attenuate alcoholic fatty liver by up-regulating PPAR-alpha and insulin signaling pathways while down-regulating SREBP-1 activity and suppressing TNF-alpha production. Recent studies show that paracrine activation of hepatic cannabinoid receptor 1 by hepatic stellate cell-derived endocannabinoids also contributes to the development of alcoholic fatty liver. Furthermore, oxidative modifications and inactivation of the enzymes involved in the mitochondrial and/or peroxisomal beta-oxidation of fatty acids could contribute to fat accumulation in the liver.


Neuronal intrinsic properties shape naturally evoked sensory inputs in the dorsal horn of the spinal cord.
Intrinsic electrophysiological properties arising from specific combinations of voltage-gated channels are fundamental for the performance of small neural networks in invertebrates, but their role in large-scale vertebrate circuits remains controversial. Although spinal neurons have complex intrinsic properties, some tasks produce high-conductance states that override intrinsic conductances, minimizing their contribution to network function. Because the detection and coding of somato-sensory information at early stages probably involves a relatively small number of neurons, we speculated that intrinsic electrophysiological properties are likely involved in the processing of sensory inputs by dorsal horn neurons (DHN). To test this idea, we took advantage of an integrated spinal cord-hindlimbs preparation from turtles allowing the combination of patch-clamp recordings of DHN embedded in an intact network, with accurate control of the extracellular milieu. We found that plateau potentials and low threshold spikes (LTS) -mediated by L- and T-type Ca(2+)channels, respectively- generated complex dynamics by interacting with naturally evoked synaptic potentials. Inhibitory receptive fields could be changed in sign by activation of the LTS. On the other hand, the plateau potential transformed sensory signals in the time domain by generating persistent activity triggered on and off by brief sensory inputs and windup of the response to repetitive sensory stimulation. Our findings suggest that intrinsic properties dynamically shape sensory inputs and thus represent a major building block for sensory processing by DHN. Intrinsic conductances in DHN appear to provide a mechanism for plastic phenomena such as dynamic receptive fields and sensitization to pain.


Peripherally Induced Regulatory T Cells: Recruited Protectors of the Central Nervous System against Autoimmune Neuroinflammation.
Defects in regulatory T cells (Treg cells) aggravate multiple sclerosis (MS) after its onset and the absence of Treg cell functions can also exacerbate the course of disease in an animal model of MS. However, autoimmune neuroinflammation in many MS models can be acutely provoked in healthy animals leading to an activation of encephalitogenic T cells despite the induction of immune tolerance in the thymus including thymically produced (t)Treg cells. In contrast, neuroinflammation can be ameliorated or even completely prevented by the antigen-specific Treg cells formed extrathymically in the peripheral immune system (pTreg cells) during tolerogenic responses to relevant neuronal antigens. This review discusses the specific roles of Treg cells in blocking neuroinflammation, examines the impact of peripheral tolerance and dendritic cells on a relevant regulation of neuroinflammation, and explores some of the most recent advances in elucidation of specific mechanisms of the conversion and function of pTreg cells including the roles of CD5 and Hopx in these processes.


Associations between infant growth and pubertal onset timing in a multiethnic prospective cohort of girls.
Early puberty increases risk of adverse health conditions throughout the life course. US girls are experiencing earlier puberty without clear reasons. Studies suggest early life factors, such as infant growth, may influence pubertal timing. We assessed the associations between infant growth and onset of breast development (thelarche), pubic hair development (pubarche), and menarche in girls. A prospective cohort of girls born at a Kaiser Permanente Northern California medical facility in 2005-11 was used. Weight-for-age z-scores were calculated at birth and 24 months. Difference in z-scores greater than 0.67 represent rapid "catch-up" growth, less than -0.67 represent delayed "catch-down" growth, and between -0.67 and 0.67 represent "normal" growth. Pubertal onset was measured using clinician-assessed sexual maturity ratings (SMRs) and defined as the age at transition from SMR 1 to SMR 2 + for both thelarche and pubarche. SMR data was collected through June 2020. Menarche was analyzed as a secondary outcome. Weibull and modified Poisson regression models were used. Models were adjusted for potential confounders. There were 15,196 girls included in the study. Approximately 30.2% experienced catch-up growth, 25.8% experienced catch-down growth, and 44% had normal growth. Girls with catch-up growth had increased risk of earlier thelarche (hazard ratio = 1.26, 95% confidence interval (CI): 1.18, 1.35), pubarche (1.38, 95% CI: 1.28, 1.48), and menarche (< 12y, relative risk = 1.52, 95% CI: 1.36, 1.69) compared to those with normal growth, after adjusting for covariates. These associations were partially mediated by childhood body mass index. Catch-down growth was associated with later pubertal onset. Girls who experience infant catch-up growth have higher risk of earlier pubertal development compared to girls with normal growth and the associations are partially explained by childhood obesity. This information may help clinicians to monitor girls who are at high risk of developing earlier.


Long-term Outcomes in Intestinal Rehabilitation in Children.
Long-term outcomes for pediatric patients with intestinal failure have significantly improved with advances in management of parenteral nutrition and the associated comorbidities. These changes have been driven by the development of multidisciplinary intestinal rehabilitation teams. Overall survival and transplant-free survival rates have increased while the introduction of new management strategies has decreased complications such as central line infections and intestinal failure associated liver disease. Factors have been identified that aid in prediction of duration of parenteral nutrition and time to enteral autonomy. Close long-term monitoring of growth and early evaluation of development are needed as abnormalities in both areas are common. With the improved survival, an important focus going forward will be the study of and improvement in quality of life for both children with intestinal failure and their families.


Taking the leap earlier: the timing of tolerance.
The paradigm for the primary prevention of peanut allergy was, until recently, strict avoidance of peanut during pregnancy, infancy, and early childhood. However, in countries that had these dietary guidelines, the prevalence of peanut allergy greatly increased in contrast to low prevalence rates of peanut allergy in countries where infant diets included peanut at an early age, suggesting the hypothesis that early introduction of peanut is superior to delayed introduction of peanut as a strategy for the primary prevention of food allergy. This review examines recent high-quality studies that support this hypothesis. Publication of the Learning Early about Allergy to Peanut interventional trial showed that 17% of children avoiding peanut until 5 years of age developed peanut allergy compared with 3% of children who started eating peanut at ages of 4-11 months. Two birth cohort studies demonstrated that high maternal consumption of peanut during the pregnancy period reduced the risk of peanut allergy in offspring by 50-75%. Recent studies indicate that early introduction of peanut enhances the development of tolerance to peanut, reducing the risk of childhood peanut allergy.


Prior-image-based CT reconstruction using attenuation-mismatched priors.
Prior-image-based reconstruction (PIBR) methods are powerful tools for reducing radiation doses and improving the image quality of low-dose computed tomography (CT). Apart from anatomical changes, prior and current images can also have different attenuations because they originated from different scanners or from the same scanner but with different x-ray beam qualities (e.g., kVp settings, beam filters) during data acquisition. In such scenarios, with attenuation-mismatched priors, PIBR is challenging. In this work, we investigate a specific PIBR method, called statistical image reconstruction, using normal-dose image-induced nonlocal means regularization (SIR-ndiNLM), to address PIBR with such attenuation-mismatched priors and achieve quantitative low-dose CT imaging. We propose two corrective schemes for the original SIR-ndiNLM method, (1) a global histogram-matching approach and (2) a local attenuation correction approach, to account for the attenuation differences between the prior and current images in PIBR. We validate the efficacy of the proposed schemes using images acquired from dual-energy CT scanners to simulate attenuation mismatches. Meanwhile, we utilize different CT slices to simulate anatomical mismatches or changes between the prior and the current low-dose image. We observe that the original SIR-ndiNLM introduces artifacts to the reconstruction when an attenuation-mismatched prior is used. Furthermore, we find that a larger attenuation mismatch between the prior and current images results in more severe artifacts in the SIR-ndiNLM reconstruction. Our two proposed corrective schemes enable SIR-ndiNLM to effectively handle the attenuation mismatch and anatomical changes between the two images and successfully eliminate the artifacts. We demonstrate that the proposed techniques permit SIR-ndiNLM to leverage the attenuation-mismatched prior and achieve quantitative low-dose CT reconstruction from both low-flux and sparse-view data acquisitions. This work permits robust and reliable PIBR for CT data acquired using different beam settings.


Enzyme-Instructed Assembly and Disassembly Processes for Targeting Downregulation in Cancer Cells.
Cancer cells differ from normal cells in both gain of functions (i.e., upregulation) and loss of functions (i.e., downregulation). While it is common to suppress gain of function for chemotherapy, it remains challenging to target downregulation in cancer cells. Here we show the combination of enzyme-instructed assembly and disassembly to target downregulation in cancer cells by designing peptidic precursors as the substrates of both carboxylesterases (CESs) and alkaline phosphatases (ALPs). The precursors turn into self-assembling molecules to form nanofibrils upon dephosphorylation by ALP, but CES-catalyzed cleavage of the ester bond on the molecules results in disassembly of the nanofibrils. The precursors selectively inhibit the cancer cells that downregulate CES (e.g., OVSAHO) but are innocuous to a hepatocyte that overexpresses CES (HepG2), while the two cell lines exhibit comparable ALP activities. This work illustrates a potential approach for the development of chemotherapy via targeting downregulation (or loss of functions) in cancer cells.


Rational design of small-molecule responsive protein switches.
Small-molecule responsive protein switches are powerful tools for controlling cellular processes. These switches are designed to respond rapidly and specifically to their inducer. They have been used in numerous applications, including the regulation of gene expression, post-translational protein modification, and signal transduction. Typically, small-molecule responsive protein switches consist of two proteins that interact with each other in the presence or absence of a small molecule. Recent advances in computational protein design already contributed to the development of protein switches with an expanded range of small-molecule inducers and increasingly sophisticated switch mechanisms. Further progress in the engineering of small-molecule responsive switches is fueled by cutting-edge computational design approaches, which will enable more complex and precise control over cellular processes and advance synthetic biology applications in biotechnology and medicine. Here, we discuss recent milestones and how technological advances are impacting the development of chemical switches.


Differential presence of fibrillation in mimic muscles in Bell's palsy.
The presence of fibrillation in mimic muscles in Bell's palsy was investigated during a clinical and electrophysiological follow-up study. Fibrillation was present in 58,82% of the frontalis and mentalis, but only in 29,41% of the orbicularis oculi (OO) muscles tested. Furthermore, the amount and the time course of fibrillation were also different, without any significant differences concerning the EMG pattern among the three muscles. The compound muscle action potential (CMAP) of the OO was negatively correlated with the presence of fibrillation. The recovery of OO was slightly better than that of the others muscles. An attempt of interpretation of these findings is made on the line of a possible sparing of the OO nerve fibers and the unique histochemical properties of the OO. On these grounds, fibrillation in the OO in Bell's palsy is less frequent than as in the other mimic muscles, and its absence is not necessarily indicating a good prognosis.


Strategies for public health adaptation to climate change in practice: social learning in the processionary Moth Knowledge Platform.
Social learning theory can support understanding of how a group of diverse actors addresses complex challenges related to public health adaptation. This study focuses on one specific issue of public health adaptation: oak processionary moth (OPM) adaptation. With a social learning framework, we examined how public health adaption strategies gradually develop and are adjusted on the basis of new knowledge and experiences. For this qualitative case study, data were collected through 27 meetings of the Processionary Moth Knowledge Platform in the Netherlands and six additional interviews. Results indicate that relations between stakeholders, including experts played a major role in the learning process, facilitating the development and implementation of OPM adaptation and connecting local challenges to national adaptation strategies. Uncertainties regarding knowledge and organization were recurrent topics of discussion, highlighting the iterative and adaptive nature of public health adaptation. The study emphasizes the importance of building relationships among stakeholders and small steps in the learning process that can lead to the creation of new strategies and, if successful, the prevention of negative health impacts.


Mechanisms of antigen processing.
Using MAb and monovalent Fab probes and saponin permeabilization we have demonstrated that PEC and TA3 B lymphoma-hybridoma cells contain a significant intracellular pool of Ia. At least in TA3 cells, this intracellular pool was independent of protein synthesis. In PEC, adherence caused redistribution of Ia with disappearance of the intracellular pool. Endocytosis of Ia occurred in both TA3 and PEC, and internalized Ia reached a plateau level corresponding in size to the total intracellular Ia pool revealed by saponin treatment. These results suggest that intracellular Ia is largely in a recycling pool derived from the plasma membrane by endocytosis. Subcellular fractionation studies suggest that Ia processing occurs in endosomes similar to those involved in transferrin processing. Antigen processing by TA3 cells was found to be unaffected by cycloheximide. In contrast, antigen processing by adherent PEC was markedly inhibited by cycloheximide, despite the fact that they maintained surface Ia and were still capable of presenting antigen peptides. This suggests that an important intracellular Ia processing step or antigen processing step was blocked in these cells. Adherent PEC may contain less recycling Ia, making protein synthesis the major source for intracellular Ia and the availability of intracellular Ia sensitive to cycloheximide. Alternatively, the inhibition of antigen processing by cycloheximide in PEC may reflect depletion of enzymes or other factors involved in antigen processing. Proteins and polysaccharides may interfere with the events that result in the formation of an immunogenic Ia-peptide complex. We had previously documented that peptides compete for the binding site of Ia molecules. We discussed here a second form of interference by polysaccharides and microbial products. These materials did not compete or interfere with the binding and presentation of processed peptides by Ia. Rather, their presence inside the macrophage inhibited MHC-dependent presentation of immunogenic proteins by inhibiting intracellular steps in antigen processing. This intracellular interference with antigen presentation can be of major importance in the presentation of complex mixtures of protein and carbohydrates.


Measuring Habitual Physical Activity in Neuromuscular Disorders: A Systematic Review.
Free-living or habitual physical activity (HPA) refers to someone's performance in his or her free-living environment. Neuromuscular disorders (NMD) manifest through HPA, and the observation of HPA can be used to identify clinical risks and to quantify outcomes in research. This review summarizes and analyses previous studies reporting the assessment of HPA in NMD, and may serve as the basis for evidence-based decision-making when considering assessing HPA in this population. A systematic review was performed to identify all studies related to HPA in NMD, followed by a critical appraisal of the assessment methodology and a final review of the identified HPA tools. A total of 22 studies were selected, reporting on eight different direct tools (or activity monitors) and ten structured patient-reported outcomes. Overall, HPA patterns in NMD differ from healthy control populations. There was a noticeable lack of validation studies for these tools and outcome measures in NMD. Very little information regarding feasibility and barriers for the application of these tools in this population have been published. The variety and heterogeneity of tools and methods in the published literature makes the comparison across different studies difficult, and methodological guidelines are warranted. We propose a checklist of considerations for the assessment and reporting of HPA in NMD.


First report of aphaenopsoid trechines (Coleoptera: Carabidae: Trechini) from Serbia, with descriptions of new taxa.
The trechine ground beetle taxa, Velesaphaenops gen. n., Velesaphaenops tarensis sp. n., and Acheroniotes lethensis sp. n., are described and diagnosed. The new taxa differ clearly from their closest relatives in a number of external characters and the shape of the genitalia. They probably belong to phyletic lineages of Pliocene age (the age of the palaeokarst of Kamena Gora and Mt. Tara). The new taxa are endemic to western and southwestern Serbia. Keys to the aphaenopsoid trechine genera in Serbia and to species of the genus Acheroniotes Lohaj Lakota, 2010 are appended.


HLA class III genes involvement in Kawasaki disease: a case-control study in Caucasian population.
Kawasaki disease (KD) is an acute, multisystemic, febrile vasculitis of unknown aetiology, which affects young children mainly under 5 years of age. The clinical variability has until now prevented to decrypt KD aetiological factors. Recently, the importance of genetics and the pivotal role of the immune system have emerged. To investigate in this direction, genomic DNA from 74 Caucasian KD cases and 440 healthy controls has been analysed to characterize functional polymorphisms of relevant HLA class III genes: AGER -429 and -374, TNF -857, -308 and -238, HSPA1A +190, HSPA1B +1267 and HSPA1L +2437. Allele, genotype and haplotype frequencies were therefore compared with the chi-squared test and Fisher's exact test. Our data showed significant deviations between patients with Kawasaki disease and controls concerning the TNF -308 polymorphism genotype (GG: P = 0.0449) and allele (G,A: P = 0.0433) and -238 polymorphism genotype frequencies (AA: P = 0.0351). Moreover, we found differences concerning the HSPA1A +190 polymorphism (GC: P = 0.0317) and the HSPA1L +2437 polymorphism (TT: P = 0.0072; TC: P = 0.0250; T: P = 0.0037; C: P = 0.0037). The calculation of TNF -238 and HSPA1L haplotype frequencies also pointed out a statistically significant decrease in patients of CG haplotype (P = 0.0001), which could have a role in protecting from the inflammatory processes that characterize the disease progression. The results obtained point to a possible involvement of the entire HLA class III region in KD susceptibility.


Esx Paralogs Are Functionally Equivalent to ESX-1 Proteins but Are Dispensable for Virulence in Mycobacterium marinum.
Mycobacterium marinum is a nontuberculous pathogen of poikilothermic fish and an opportunistic human pathogen. Like tuberculous mycobacteria, the M. marinum M strain requires the ESX-1 (ESAT-6 system 1) secretion system for virulence in host cells. EsxB and EsxA, two major virulence factors exported by the ESX-1 system, are encoded by the esxBA genes within the ESX-1 locus. Deletion of the esxBA genes abrogates ESX-1 export and attenuates M. marinum in ex vivo and in vivo models of infection. Interestingly, there are several duplications of the esxB and esxA genes (esxB_1, esxB_2, esxA_1, esxA_2, and esxA_3) in the M. marinum M genome located outside the ESX-1 locus. We sought to understand if this region, known as ESX-6, contributes to ESX-1-mediated virulence. We found that deletion of the esxB_1 gene alone or the entire ESX-6 locus did not impact ESX-1 export or function, supporting the idea that the esxBA genes present at the ESX-1 locus are the primary contributors to ESX-1-mediated virulence. Nevertheless, overexpression of the esxB_1 locus complemented ESX-1 function in the ΔesxBA strain, signifying that the two loci are functionally equivalent. Our findings raise questions about why duplicate versions of the esxBA genes are maintained in the M. marinum M genome and how these proteins, which are functionally equivalent to virulence factors, contribute to mycobacterial biology.IMPORTANCEMycobacterium tuberculosis is the causative agent of the human disease tuberculosis (TB). There are 10.4 million cases and 1.7 million TB-associated deaths annually, making TB a leading cause of death globally. Nontuberculous mycobacteria (NTM) cause chronic human infections that are acquired from the environment. Despite differences in disease etiology, both tuberculous and NTM pathogens use the ESX-1 secretion system to cause disease. The nontubercular mycobacterial species, Mycobacterium marinum, has additional copies of specific ESX-1 genes. Our findings demonstrate that the duplicated genes do not contribute to virulence but can substitute for virulence factors in M. marinum These findings suggest that the duplicated genes may play a specific role in NTM biology.


On-chip enzyme quantification of single Escherichia coli bacteria by immunoassay-based analysis.
Individual bacteria of an isogenic population can differ significantly in their phenotypic characteristics. This cellular heterogeneity is thought to increase the adaptivity to environmental changes on a population level. Analytical methods for single-bacteria analyses are essential to reveal the different factors that may contribute to this cellular heterogeneity, among them the stochastic gene expression, cell cycle stages and cell aging. Although promising concepts for the analysis of single mammalian cells based on microsystems technology were recently developed, platforms suitable for proteomic analyses of microbial cells are by far more challenging. Here, we present a microfluidic device optimized for the analysis of single Escherichia coli bacteria. Individual bacteria are captured in a trap and isolated in a volume of only 155 pL. In combination with an immunoassay-based analysis of the cell lysate, the platform allowed the selective and sensitive analysis of intracellular enzymes. The limit of detection of the developed protocol was found to be 200 enzymes. Using this platform, we could investigate the levels of β-galactosidase in cells grown under different nutrient conditions. We successfully determined the enzyme copy numbers in cells cultured in defined medium (3517 ± 1578) and in complex medium (4710 ± 2643), and verified the down-regulation of expression in medium that contained only glucose as carbon source. The strong variations we found for individual bacteria confirm the phenotype heterogeneity. The capability to quantify proteins and other molecules in single bacterial lysates is encouraging to use the new analysis platform in future proteomics studies of isogenic bacteria populations.


The effect of chronic periodontal disease on human jaw muscles: a pilot study using computed tomography.
Patients with chronic periodontal disease sometimes complain of difficulty in masticating hard foods. Computed tomography (CT) was used to investigate the cross-sectional area (CSA) and density of the masseter and medial pterygoid muscles in a pilot study of 12 healthy female subjects aged 30-39 years. Six subjects (the 'diseased' group) had suffered from chronic periodontal disease and had, on average, 40% bone loss. Their disease had been treated by scaling and root planning and had remained clinically stable for at least 6 months before their inclusion in the study. The six age-matched 'control' subjects had no history of periodontal disease. All bone-loss measurements were evaluated as a percentage of root length. At defined scan planes, sections of both muscles were readily identified. Results from the 'diseased' group as compared with the 'control' group revealed that in the former the CSA of masseter and medial pterygoid was reduced significantly by 25% and 10%, respectively. There were no significant differences in the X-ray density of both muscles between the two groups, which indicated a similar fibro-fatty content. These findings indicate a reduction in the masticatory force that may be utilized by patients with reduced periodontal support and may help explain their masticatory difficulties.


Preliminary report of a mathematical model of ventilation and intrathoracic pressure applied to prehospital patients with severe traumatic brain injury.
Inadvertent hyperventilation is associated with poor outcomes from traumatic brain injury (TBI). Hypocapnic cerebral vasoconstriction is well described and causes an immediate and profound decrease in cerebral perfusion. The hemodynamic effects of positive-pressure ventilation (PPV) remain incompletely understood but may be equally important, particularly in the hypovolemic patient with TBI. Preliminary report on the application of a previously described mathematical model of perfusion and ventilation to prehospital data to predict intrathoracic pressure. Ventilation data from 108 TBI patients (76 ground transported, 32 helicopter transported) were used for this analysis. Ventilation rate (VR) and end-tidal carbon dioxide (PetCO2) values were used to estimate tidal volume (VT). The values for VR and estimated VT were then applied to a previously described mathematical model of perfusion and ventilation. This model allows input of various lung parameters to define a pressure-volume relationship, then derives mean intrathoracic pressure (MITP) for various VT and VR values. For this analysis, normal lung parameters were utilized. Separate analyses were performed assuming either fixed or variable PaCO2-PetCO2 differences. Ground and air medical patients were compared with regard to VR, PetCO2, estimated VT, and predicted MITP. A total of 10,647 measurements were included from the 108 TBI patients, representing about 13 minutes of ventilation per patient. Mean VR values were higher for ground patients versus air patients (21.6 vs. 19.7 breaths/min; p < 0.01). Estimated VT values were similar for ground and air patients (399 mL vs. 392 mL; p = NS) in the fixed model but not the variable (636 vs. 688 mL, respectively; p < 0.01). Mean PetCO2 values were lower for ground versus air patients (30.6 vs. 33.8 mmHg; p < 0.01). Predicted MITP values were higher for ground versus air patients, assuming either fixed (9.0 vs. 8.1 mmHg; p < 0.01) or variable (10.9 vs. 9.7 mmHg; p < 0.01) PaCO2-PetCO2 differences. Predicted MITP values increased with ventilation rates. Future studies to externally validate this model are warranted.


Characterization of ApB73, a virulence factor important for colonization of Zea mays by the smut Ustilago maydis.
The biotrophic fungus Ustilago maydis, the causal agent of corn smut disease, uses numerous small secreted effector proteins to suppress plant defence responses and reshape the host metabolism. However, the role of specific effectors remains poorly understood. Here, we describe the identification of ApB73 (Apathogenic in B73), an as yet uncharacterized protein essential for the successful colonization of maize by U. maydis. We show that apB73 is transcriptionally induced during the biotrophic stages of the fungal life cycle. The deletion of the apB73 gene results in cultivar-specific loss of gall formation in the host. The ApB73 protein is conserved among closely related smut fungi. However, using virulence assays, we show that only the orthologue of the maize-infecting head smut Sporisorium reilianum can complement the mutant phenotype of U. maydis. Although microscopy shows that ApB73 is secreted into the biotrophic interface, it seems to remain associated with fungal cell wall components or the fungal plasma membrane. Taken together, the results show that ApB73 is a conserved and important virulence factor of U. maydis that localizes to the interface between the pathogen and its host Zea mays.


Comparing the effect of e-learning and educational booklet on the childbirth self-efficacy: a randomized controlled clinical trial.
This study aimed to compare the effect of e-learning and educational booklet on the childbirth self-efficacy (CBSE). This randomized controlled clinical trial was conducted on 153 pregnant women referred to health centers in the city of Miandoab, Iran in 2015-2016. Participants were assigned into two intervention groups (e-learning and educational booklet) and the control group. A single face-to-face session was held for intervention groups about the management of labor pain in 30-34 weeks of pregnancy and the booklet and software were provided. The CBSE questionnaire was filled out by the participants before intervention and active phase of labor at 4-5 cm dilatation of cervix. One-way ANOVA and ANCOVA test with adjusting the baseline scores were used to compare the mean score of self-efficacy among study groups respectively before and after the intervention. There was no significant difference between the three groups in terms of socio-demographic characteristics (p > 0.05). After the intervention, the mean score of the CBSE in the educational booklet group (adjusted mean difference: 113.4; confidence interval 95%: 100.7-126.1) and e-learning group (159.3; 146.5-172.0) was significantly higher than the control group. Also, the mean score of the CBSE in the e-learning group had a significant increase compared to the educational booklet group (45.9; 33.0-58.7). The results indicate that e-learning and educational booklet are effective in enhancing mothers' CBSE. Thus, the mothers are recommended to use these teaching methods.


Mating structure, isonymy and social class in late nineteenth century Tasmania.
This study analysed all marriages (N = 4258) registered in the island State of Tasmania during the period 1896-1899 for mating structure, using marital isonymy. We found that irrespective of the geographic size of the subdivisions the random component (Fr) values were similar. This is because there was a high proportion of unique names despite the large population size. Fn values were sensitive to geographic distance only with respect to the 'unrealistically' larger units of analysis. Isonymy is valid over very large distances in Tasmania. Inbreeding values in the study were typical of other mainstream populations, but the nonrandom component was unusually high. Farmer marriages were the major contributors to the high Fn values. One particular region, the Midlands, displayed both a high F (0.0081) and Fn (0.0074) component. The explanation of these values lies in the effects of a rigid socio-economic structure established during colonization.


Revised Theoretical Model on Enantiocontrol in Phosphoric Acid Catalyzed H-Transfer Hydrogenation of Quinoline.
The enantioselective H-transfer hydrogenation of quinoline by Hantzsch ester is a relevant example of Brønsted acid catalyzed cascade reactions, with phosphoric acid being a privileged catalyst. The generally accepted mechanism points out the hydride transfer step as the rate- and stereodetermining step, however computations based on these models do not totally fit with experimental observations. We hereby present a computational study that enlightens the stereochemical outcome and quantitatively reproduces the experimental enantiomeric excesses in a series of H-transfer hydrogenations. Our calculations suggest that the high stereocontrol usually attained with BINOL-derived phosphoric acids results mostly from the steric constraints generated by an aryl substituent of the catalyst, which hinders the access of the Hantzsch ester to the catalytic site and enforces approach through a specific way. It relies on a new model involving the preferential assembly of one of the stereomeric complexes formed by the chiral phosphoric acid and the two reaction partners. The stereodetermining step thus occurs prior to the H-transfer step.


Agonism of 4-1BB for immune therapy: a perspective on possibilities and complications.
Costimulatory receptors on immune cells represent attractive targets for immunotherapy given that these molecules can increase the frequency of individual protective immune cell populations and their longevity, as well as enhance various effector functions. 4-1BB, a member of the TNF receptor superfamily, also known as CD137 and TNFRSF9, is one such molecule that is inducible on several cell types, including T cells and NK cells. Preclinical studies in animal models have validated the notion that stimulating 4-1BB with agonist reagents or its natural ligand could be useful to augment conventional T cell and NK cell immunity to protect against tumor growth and against viral infection. Additionally, stimulating 4-1BB can enhance regulatory T cell function and might be useful in the right context for suppressing autoimmunity. Two human agonist antibodies to 4-1BB have been produced and tested in clinical trials for cancer, with variable results, leading to the production of a wealth of second-generation antibody constructs, including bi- and multi-specifics, with the hope of optimizing activity and selectivity. Here, we review the progress to date in agonism of 4-1BB, discuss the complications in targeting the immune system appropriately to elicit the desired activity, together with challenges in engineering agonists, and highlight the untapped potential of manipulating this molecule in infectious disease and autoimmunity.


Alternative splicing of human papillomavirus type-16 E6/E6* early mRNA is coupled to EGF signaling via Erk1/2 activation.
Certain types of human papillomaviruses (HPVs) are etiologically linked to cervical cancer. Their transforming capacity is encoded by a polycistronic premRNA, where alternative splicing leads to the translation of functional distinct proteins such as E6, E6*, and E7. Here we show that splicing of HPV16 E6/E7 ORF cassette is regulated by the epidermal growth factor (EGF) pathway. The presence of EGF was coupled to preferential E6 expression, whereas depletion of EGF, or treatment with EGF receptor (EGFR) neutralizing antibodies or the EGFR inhibitor tyrphostin AG1478, resulted in E6 exon exclusion in favor of E6*. As a consequence, increased p53 levels and enhanced translation of E7 with a subsequent reduction of the retinoblastoma protein pRb could be discerned. E6 exon exclusion upon EGF depletion was independent from promoter usage, mRNA stability, or selective mRNA transport. Time-course experiments and incubation with cycloheximide demonstrated that E6 alternative splicing is a direct and reversible effect of EGF signal transduction, not depending on de novo protein synthesis. Within this process, Erk1/2-kinase activation was the critical event for E6 exon inclusion, mediated by the upstream MAP kinase MEK1/2. Moreover, siRNA knockdown experiments revealed an involvement of splicing factors hnRNPA1 and hnRNPA2 in E6 exon exclusion, whereas the splicing factors Brm and Sam68 were found to promote E6 exon inclusion. Because there is a natural gradient of EGF and EGF receptor expression in the stratified epithelium, it is reasonable to assume that EGF modulates E6/E7 splicing during the viral life cycle and transformation.


Variation in the fine structure of a marine achromobacter and a marine pseudomonad grown under selected nutritional and temperature regimes.
Certain features of the fine structure of a marine achromobacter and a marine pseudomonad were dependent upon the conditions of growth. Cells of achromobacter grown at 10 C in a low peptone-seawater (SW) medium displayed the characteristic morphology of the achromobacter: a regularly undulant outer element of the cell wall and a planar inner element, tightly packed ribonucleoprotein (RNP) particles in the cytoplasm, deoxyribonucleic acid (DNA) disposed in a lobate manner, and dense inclusion bodies. Few mesosomes, however, were seen. Cells of achromobacter grown at 10 C in a high peptone-SW medium had larger and more highly organized mesosomes. At 22 C, in a low peptone-SW medium, no mesosomes were seen, but the inclusions were more frequently seen and were larger in the achromobacter cells. At 22 C, in a high peptone-SW medium, these cells revealed the greatest variation in cellular morphology. They contained both small and large mesosomes, or no mesosomes, and both small and large inclusions, or no inclusions. Pseudomonad cells at 10 C in a low peptone-SW medium revealed a typical gram-negative morphology: double-layered, irregularly undulant cell wall; more nearly planar cytoplasmic membrane; densely stained, lightly packed RNP particles; finely fibrillar, axially disposed DNA; simple mesosomes. At 10 C, in a high peptone-SW medium, pseudomonad cells revealed associated strands of material and intracytoplasmic ringlike structures. At 22 C, in a low peptone-SW medium, pseudomonad cells had a more undulant cell-wall and a more nearly planar cytoplasmic membrane. At 22 C, in a high peptone-SW medium, these cells revealed prominent blebs of the cell wall.


Environmental and genetic sources of familial aggregation of blood pressure in Tecumseh, Michigan.
Environmental and genetic sources of variation in blood pressure have been assessed for an 82% sample of the Tecumseh, Michigan, population during the period 1962-1965. Correlations of systolic blood pressure and diastolic blood pressure were calculated for sets of related and unrelated individuals living together and related individuals living apart. These correlations were analyzed using a biologic model which partitions the total phenotypic variance into components associated with shared genes and components associated with shared household environments both within and across generations. Heritabilities were estimated to be 0.42 and 0.30 for systolic blood pressure and diastolic blood pressure, respectively. The model confirms the hypothesis that the determinants of blood pressure consist of both significant genetic (p less than 0.005) and environmental (p less than 0.005) components. However, only the shared household environments within generations were found to be significant for systolic blood pressure (p less than 0.005) and diastolic blood pressure (p less than 0.05). Although not significant, the across generation shared environment between mothers and offspring tended to have some importance, while that between fathers and offspring was negligible. The implication that there may be some maternal influence in addition to the genetic influence on the determination of blood pressure in offspring warrants further investigation.


[The role of food allergy as a provoking factor of migraine].
To assess the effect of food allergies on the course of migraine. Seventy patients with migraine, aged 21-56 years old, were examined using headache diary, MIDAS and VAS, studies of specific antibodies of the IgG4 class (delayed type food allergy) by immuno-enzyme analysis (ELISA), microbiological examination of a smear from the mucous membrane of the posterior wall of the oropharynx with mass spectrometry of microbial markers (MSMM) with the identification of 57 microorganisms. We found an increase in specific IgG4 for a number of food allergens in most patients with migraine, of which 48.5% had a pronounced increase in IgG4 (>150 kEd/l) for at least one allergen (cow's milk - 13% patients, wheat flour - 5%, egg white - 47% or yolk - 26%, quail egg - 15%, sweet pepper - 6%), in 29% of people to several food allergens at once (all of them had chicken egg protein as one of the allergens). There was the association of IgG4 titers to wheat allergen with the severity of headache according to VAS (r-S=0.7; p=0.0046) in patients with the most severe, chronic migraine (17 people) and with an imbalance of the oropharyngeal microbiota, namely, concentration of pathological viruses Herpes spp. (rs=0.29; p=0.02), Epstein-Barr (rs=0.46; p=0.0002) and microscopic fungi (rs=0.39; p=0.0016), detected in these patients. We show for the first time the relationship between delayed-type food allergy and redistribution in the microbiome of the oropharynx of patients with migraine and once again confirm the role of delayed-type food allergy as a clinically significant factor influencing the course of migraine (its intensity and chronicity).


Vitamin D: a negative acute phase reactant.
We evaluated the effect of the systemic inflammatory response (SIR), as provoked by elective orthopaedic surgery, on serum vitamin D [25-(OH)D]. Serum 25-(OH)D, serum vitamin D binding protein (VDBP) and urinary VDBP were measured in 30 patients before and 48-hours after knee or hip arthroplasty. C-reactive protein (CRP) was measured to assess the SIR. The mean (SD) CRP increased following surgery [5.0 (5.5) vs 116.0 (81.2) mg/L; P<0.0001] as did urine VDBP/Creatinine ratio [8 (9) vs 20 (25) pg/mmol; p=0.0004]. Serum 25-(OH)D [56.2 (30.3) vs 46.0 (27.6) nmol/L; p = 0.0006] and serum VDBP [334 (43) vs 298 (37) mg/L]; P<0.0001] decreased. Serum 25-(OH)D is a negative acute phase reactant, which has implications for acute and chronic inflammatory diseases. Serum 25-(OH)D is an unreliable biomarker of vitamin D status after acute inflammatory insult. Hypovitaminosis D may be the consequence rather than cause of chronic inflammatory diseases.


The effect of medetomidine on GABA and benzodiazepine receptors in vivo: lack of anxiolytic but some evidence of possible stress-protective activity.
Medetomidine, a new selective alpha 2-adrenoceptor agonist, potentiated bicuculline seizures in mice. In vivo pretreatment with medetomidine in mouse cerebral cortex reduced dose-dependently (2.5-100 micrograms/kg) GABA-potentiated 3H-flunitrazepam binding. The affinity of 3H-muscimol was also reduced by medetomidine. This effect of medetomidine on GABA-potentiated benzodiazepine binding was reversed by pretreatment with atipamezole (1 mg/kg), a specific alpha 2-antagonist. In an elevated plus-maze model of anxiety medetomidine (0.5-10 micrograms/kg) was inactive both in rats and mice and did not antagonize the behavioural effects of an anxiogenic beta-carboline, DMCM. However, at lower doses medetomidine (10 but not 50 micrograms/kg) antagonized the swimming stress caused increase of central benzodiazepine binding sites (labeled with 3H-Ro 15-1788) in mouse cerebral cortex. The increase of peripheral benzodiazepine binding sites on brain and heart cryostat cut slices caused by stress was also antagonized by pretreatment with medetomidine. The behavioural and biochemical data obtained in this study are evidence that medetomidine does not have anxiolytic effect but may have, in lower doses, stress-protective activity.


Comparison of Human Primary with Human iPS Cell-Derived Dopaminergic Neuron Grafts in the Rat Model for Parkinson's Disease.
Neuronal degeneration within the substantia nigra and the loss of the dopaminergic nigro-striatal pathway are the major hallmarks of Parkinson's disease (PD). Grafts of foetal ventral mesencephalic (VM) dopaminergic (DA) neurons into the striatum have been shown to be able to restore striatal dopamine levels and to improve overall PD symptoms. However, human foetus-derived cell grafts are not feasible for clinical application. Autologous induced pluripotent stem cell (iPS cell)-derived DA neurons are emerging as an unprecedented alternative. In this review, we summarize and compare the efficacy of human iPS cell-derived DA neuron grafts to restore normal behaviour in a rat model for PD with that of human foetal primary DA neurons. The differences we observed in the efficacy to restore normal function between the 2 types of DA neuron grafts could be ascribed to intrinsic properties of the iPS cell-derived DA neurons that critically affected survival and proper neurite extension in the striatum after implantation.


The Use of Echocardiograms in Preparticipation Examinations.
Sudden cardiac death (SCD) is the leading cause of death during exercise in young athletes. Preparticipation physical examinations were developed to maintain the health and safety of athletes, including the prompt identification of those at risk for SCD. The use of medical history and physical examinations, electrocardiography, and echocardiography, or some combination thereof, is the source of continued debate. This article provides an overview of the etiology of SCD and reviews literature relating to preparticipation echocardiography, with a focus on its evolution, utility, and effectiveness. The limited echocardiogram is a potentially viable screening option yet to be thoroughly explored by experts and policymakers in the sports medicine community.


Nonlocal Patch Tensor Sparse Representation for Hyperspectral Image Super-Resolution.
This paper presents a hypserspectral image (HSI) super-resolution method which fuses a low-resolution hyperspectral image (LR-HSI) with a high-resolution multispectral image (HR-MSI) to get high-resolution HSI (HR-HSI). The proposed method first extracts the nonlocal similar patches to form a nonlocal patch tensor (NPT). A novel tensor-tensor product (t-product) based tensor sparse representation is proposed to model the extracted NPTs. Through the tensor sparse representation, both the spectral and spatial similarities between the nonlocal similar patches are well preserved. Then, the relationship between the HR-HSI and LR-HSI is built using t-product which allows us to design a unified objective function to incorporate the nonlocal similarity, tensor dictionary learning, and tensor sparse coding together. Finally, Alternating Direction Method of Multipliers (ADMM) is used to solve the optimization problem. Experimental results on three data sets and one real data set demonstrate that the proposed method substantially outperforms the existing state-of-the-art HSI super-resolution methods.


[Mediators of amyloidogenesis and cytokines in dialysis-related amyloidosis].
Dialysis related amyloidosis (DRA) is a severe complication of the long-term dialysis treatment. beta 2 microglobulin and probably other factors influence the development of amyloid deposits. We investigated some of these factors during hemodialysis session. We investigated 20 patients undergoing regular hemodialysis treatment. Patients were divided into AMYL group (with histologically proven DRA) and NE-AMYL group (without signs of DRA). Plasma levels of following factors were investigated using standard ELISA kits: serum amyloid A (SAA), interleukin-6 (IL-6), macrophage-colony stimulating factor (M-CSF). In addition plasma concentrations of C-reactive protein (CRP) and beta 2 microglobulin (beta 2M) were investigated in the AMYL group. All these parameters were studied during different time periods of the hemodialysis session. Plasma levels of SAA and IL-6 did not increase during hemodialysis session and we did not find any difference in plasma levels of these factors between the group of patients with AMYLand NE-AMYL. Plasma levels of M-CSF increased during hemodialysis and its levels in AMYL group were significantly higher in comparison with NE-AMYL group at the end of hemodialysis session (5345.10 +/- 340.42 vs. 3458.45 +/- 332.15 pg/ml, p = 0.0011). A linear correlation was found between plasma levels of SAA and CRP during hemodialysis whereas no correlation was found between plasma levels of beta 2M and other factors. Our study suggests that plasma levels of M-CSF are increased in patients with chronic renal failure. Significant increase of M-CSF levels in the AMYL group could lead to greater activation of monocyte-macrophage system and could serve as factor supporting amyloid deposition process.


The risk of cancer in patients with generalized anxiety disorder: a nationwide population-based study.
To evaluate the risk of cancer among patients with generalized anxiety disorder (GAD) in a nationwide population-based dataset. We recruited newly-diagnosed GAD patients aged 20 years or older without antecedent cancer from the Taiwan National Health Insurance Research database between 2000-2010. Standardized incidence ratios (SIRs) of cancers were calculated in GAD patients, and the subgroup of GAD patients diagnosed by psychiatric specialists. A total of 559 cancers developed among 19,793 GAD patients with a follow-up of 89,485 person-years (median follow-up of 4.34 years), leading to a significantly increased SIR of 1.14 [95% confidence interval (CI) 1.05-1.24]. Male GAD patients had a significantly increased SIR overall (1.30, 95% CI 1.15-1.46) and for lung and prostate cancer (1.77, 95% CI 1.33-2.30 and 2.17, 95% CI 1.56-2.93, respectively). Patients over 80 years of age also had a significantly increased SIR (1.56, 95% CI 1.25-1.92), especially in males. However, psychiatrist-diagnosed GAD patients did not show increased cancer risk relative to the general population, perhaps due to having fewer physical comorbidities than non-psychiatrist-diagnosed GAD patients. This study found that overall cancer risk is elevated among patients with GAD. The risk of lung and prostate cancer also increased in male patients with GAD. This increased cancer risk may be due to physical comorbidities and surveillance bias. Further prospective study is necessary to confirm these findings.


Aberrant lipid accumulation in the mouse visceral yolk sac resulting from maternal diabetes and obesity.
Maternal diabetes and obesity in pregnancy are well-known risk factors for structural birth defects, including neural tube defects and congenital heart defects. Progeny from affected pregnancies are also predisposed to developing cardiometabolic disease in later life. Based upon in vitro embryo cultures of rat embryos, it was postulated that nutrient uptake by the yolk sac is deficient in diabetic pregnancies. In contrast, using two independent mouse models of maternal diabetes, and a high-fat diet-feeding model of maternal obesity, we observed excessive lipid accumulation at 8.5 days in the yolk sac. The numbers as well as sizes of intracellular lipid droplets were increased in yolk sacs of embryos from diabetic and obese pregnancies. Maternal metabolic disease did not affect expression of lipid transporter proteins, including ApoA1, ApoB and SR-B1, consistent with our earlier report that expression of glucose and fatty acid transporter genes was also unchanged in diabetic pregnancy-derived yolk sacs. Colocalization of lipid droplets with lysosomes was significantly reduced in the yolk sacs from diabetic and obese pregnancies compared to yolk sacs from normal pregnancies. We therefore conclude that processing of lipids is defective in pregnancies affected by maternal metabolic disease, which may lead to reduced availability of lipids to the developing embryo. The possible implications of insufficient supply of lipids -and potentially of other nutrients-to the embryos experiencing adverse pregnancy conditions are discussed.


Total synthesis of clavatadine A.
The first total synthesis of the potent and selective human blood coagulation factor XIa inhibitor clavatadine A (1) is described. Direct, early-stage guanidinylation enabled rapid, convergent access to an immediate clavatadine A precursor. Concomitant lactone hydrolysis and guanidine deprotection with aqueous acid cleanly provided clavatadine A (1) in only four steps (longest linear sequence, 41-43% overall yield).


Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses.
It is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. In many cases of community-acquired respiratory infections, clinicians treat patients empirically. The development of molecular methods for direct detection of viruses has been progressed recently. The objective of this study was recognizing the panel of respiratory RNA viruses by multiplex SYBR Green real-time polymerase chain reaction (PCR). Randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. After RNA extraction, cDNA was synthesized. Three SYBR Green multiplex real-time PCR assays were developed for simultaneous detection of 12 respiratory RNA viruses. Each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (PIV1-4); the third set: coronaviruses NL63, 229E, severe acute respiratory syndrome (SARS), and OC43. Application of the multiplex SYBR Green real-time PCR in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) PIV3, 9 (5.23%) PIV4, and 1 (0.58%) coronavirus NL63. All the positive samples were detected during December to March (2011 - 2012). Multiplex SYBR Green real-time PCR is a rapid and relatively inexpensive method for detection of respiratory viruses.


Place-pitch discrimination of single- versus dual-electrode stimuli by cochlear implant users (L).
Simultaneous or near-simultaneous activation of adjacent cochlear implant electrodes can produce pitch percepts intermediate to those produced by each electrode separately, thereby increasing the number of place-pitch steps available to cochlear implant listeners. To estimate how many distinct pitches could be generated with simultaneous dual-electrode stimulation, the present study measured place-pitch discrimination thresholds for single- versus dual-electrode stimuli in users of the Clarion CII device. Discrimination thresholds were expressed as the proportion of current directed to the secondary electrode of the dual-electrode pair. For 16 of 17 electrode pairs tested in six subjects, thresholds ranged from 0.11 to 0.64, suggesting that dual-electrode stimuli can produce 2-9 discriminable pitches between the pitches of single electrodes. Some subjects demonstrated a level effect, with better place-pitch discrimination at higher stimulus levels. Equal loudness was achieved with dual-electrode stimuli at net current levels that were similar to or slightly higher than those for single-electrode stimuli.


The origins of nonideality exhibited by monoclonal antibodies and Fab fragments in human serum.
The development of therapeutic antibodies remains challenging, time-consuming, and expensive. A key contributing factor is a lack of understanding of how proteins are affected by complex biological environments such as serum and plasma. Nonideality due to attractive or repulsive interactions with cosolutes can alter the stability, aggregation propensity, and binding interactions of proteins in solution. Fluorescence correlation spectroscopy (FCS) can be used to measure apparent second virial coefficient (B2,app ) values for therapeutic and model monoclonal antibodies (mAbs) that capture the nature and strength of interactions with cosolutes directly in undiluted serum and similar complex biological media. Here, we use FCS-derived B2,app measurements to identify the components of human serum responsible for nonideal interactions with mAbs and Fab fragments. Most mAbs exhibit neutral or slightly attractive interactions with intact serum. Generally, mAbs display repulsive interactions with albumin and mildly attractive interactions with IgGs in the context of whole serum. Crucially, however, these attractive interactions are much stronger with pooled IgGs isolated from other serum components, indicating that the effects of serum nonideality can only be understood by studying the intact medium (rather than isolated components). Moreover, Fab fragments universally exhibited more attractive interactions than their parental mAbs, potentially rendering them more susceptible to nonideality-driven perturbations. FCS-based B2,app measurements have the potential to advance our understanding of how physiological environments impact protein-based therapeutics in general. Furthermore, incorporating such assays into preclinical biologics development may help de-risk molecules and make for a faster and more efficient development process.


Correlation between morphological and functional retinal impairment in patients affected by ocular hypertension, glaucoma, demyelinating optic neuritis and Alzheimer's disease.
In this article the correlations between the morphological evaluation of the nerve fiber layer (NFL) thickness (by OCT) and retinal functional assessment (by Pattern ERG recordings) performed in patients affected by ocular hypertension (OHT), glaucoma (OAG), demyelinating optic neuritis (MSON), and Alzheimer's disease (AD) are reported. In OHT eyes with ocular hypertension we observed that the inter-individual variation in NFL thickness is correlated with the variability of the PERG responses (the thinner the layer, the worse the visual function). In our OAG, MSON and AD eyes we observed a significant reduction in NFL thickness when compared with controls. In OHT, OAG, MSON and AD eyes abnormal PERG responses with delayed implicit times and reduced amplitudes were found. The impairment in the PERG parameters was significantly correlated to the reduction in NFL thickness. Our results suggest that in patients affected by ocular hypertension, glaucoma, demyelinating optic neuritis, and Alzheimer's Disease there is a reduction of NFL thickness evaluated "in vivo" by OCT, and this morphological involvement is correlated with electrophysiological responses assumed to be originating from the innermost retinal layers.


Practice-based evidence study design for comparative effectiveness research.
To describe a new, rigorous, comprehensive practice-based evidence for clinical practice improvement (PBE-CPI) study methodology, and compare its features, advantages, and disadvantages to those of randomized controlled trials and sophisticated statistical methods for comparative effectiveness research. PBE-CPI incorporates natural variation within data from routine clinical practice to determine what works, for whom, when, and at what cost. It uses the knowledge of front-line caregivers, who develop study questions and define variables as part of a transdisciplinary team. Its comprehensive measurement framework provides a basis for analyses of significant bivariate and multivariate associations between treatments and outcomes, controlling for patient differences, such as severity of illness. PBE-CPI studies can uncover better practices more quickly than randomized controlled trials or sophisticated statistical methods, while achieving many of the same advantages. We present examples of actionable findings from PBE-CPI studies in postacute care settings related to comparative effectiveness of medications, nutritional support approaches, incontinence products, physical therapy activities, and other services. Outcomes improved when practices associated with better outcomes in PBE-CPI analyses were adopted in practice.


Sex hormone-binding globulin: a novel marker for nodal metastases prediction in prostate cancer patients undergoing extended pelvic lymph node dissection.
To examine the association between sex hormone-binding globulin (SHBG) and lymph node invasion (LNI) in patients treated with radical retropubic prostatectomy and extended pelvic lymph node dissection (ePLND). The preoperative serum SHBG level was measured in a cohort of 168 consecutive patients (mean age 63.9 years, range 48-77) who underwent radical retropubic prostatectomy with ePLND for clinically localized prostate cancer. Logistic regression models tested the association between the predictors (including prostate-specific antigen, clinical stage, primary and secondary biopsy Gleason grades, and SHBG) and LNI. Logistic regression coefficients were used to calculate the predictive accuracy, which was subjected to 200 bootstrap resamples to reduce overfit bias. Thirteen patients (7.7%) had LNI. The mean serum SHBG level was significantly greater in the patients with LNI than in those without LNI (50.0 vs 35.1 nmol/L, respectively; P < .001). Univariate analysis indicated that preoperative SHBG was the single most informative predictor of LNI (77.8% vs 71.7% for prostate-specific antigen, 63.9% for clinical stage, and 63.1% and 54.2% for primary and secondary Gleason grade, respectively). On multivariate analysis, preoperative SHBG was still significantly associated with LNI (P < .001), after accounting for the other variables. The addition of preoperative SHBG increased the predictive accuracy of the base model using clinically established predictors from 72.7% to 82.8% (10.1% gain; P < .001). The results of this study provide novel evidence that SHBG might serve as a significant multivariate predictor of LNI in patients with prostate cancer undergoing ePLND. The use of preoperative serum SHBG could help to identify patients at risk of LNI who should undergo ePLND.


Pathophysiology of proteinuria in immune glomerular injury.
Uncharged dextrans of graded size (28-60 A) were used to evaluate glomerular barrier size selectivity in 13 nephrotic patients with diffuse proliferative lupus nephritis. Transglomerular sieving was enhanced selectively for dextrans of greater than 46 A radius when the nephritis was active and restored towards normal by immunosuppressive therapy. A mathematical model of hindered solute transport through a porous membrane revealed these findings to reflect the prevalence in the glomerular barrier of a subset of enlarged, nondiscriminatory pores. Although few in number, such enlarged pores accounted for both the magnitude and composition of observed proteinuria. We infer that impaired barrier size selectivity underlies the proteinuria that attends lupus nephritis, and that impairment of barrier charge selectivity need not be invoked in this circumstance.


Influence of carbohydrate source on liver fat accumulation in hens.
Four experiments were conducted with laying hens and one with broiler chicks to investigate the effect of carbohydrate source on fat accumulation in the liver. When corn and wheat in various proportions were fed to laying hens, the percent fat and total fat per liver increased as the proportion of corn increased. In comparison of different cereal grains and wheat samples in isocaloric diets, the total liver fat accumulated was the highest for hens fed grain sorghum, corn or triticale and the lowest for those fed barley, oats or rye. Intermediate levels of liver fat were obtained with wheat and similar liver fat contents were observed with hens fed different samples of West Coast wheat. There was no significant difference in liver fat of groups fed wheat diets made isocaloric with corn oil or animal fat, Liver fat accumulation of broiler chicks fed diets with corn or wheat was not significantly different. No difference in liver weight, liver fat content and total fat per liver was observed among hens fed diets containing either zero, one-half, or all glucose monohydrate substituted for corn. In comparisons of corn or wheat from different geographical area, no difference was observed in liver fat content of hens fed corn grown in Georgia or the Midwest or wheat grown in Georgia or in the Far West. Hens fed diets contaning corn had significantly more liver dry matter, liver fat, and total fat per liver than hens fed the isocaloric diets containing wheat. Considerably more fat accumulated in the livers of hens fed diets containing corn at Washington State University (15.4 to 19.6 g. fat/liver) than in hens fed corn at the University of Georgia (5.4 to 7.2 g. fat/liver), suggesting the existence of unidentified environmental factors affecting liver fat accumulation.


Neonicotinoid Residues in Fruits and Vegetables: An Integrated Dietary Exposure Assessment Approach.
Neonicotinoids have become the most widely used insecticides in the world since introduced in the mid 1990s, yet the extent of human exposure and health impacts is not fully understood. In this study, the residues were analyzed of seven neonicotinoids in fruit and vegetable samples collected from two cross-sectional studies: the U.S. Congressional Cafeteria study (USCC) and the Hangzhou China (HZC) study. We then employed a relative potency factor method to integrate all neonicotinoids in each food sample using the respective reference dose values as the basis for summation. The findings were compared with data published by the U.S. Department of Agriculture Pesticide Data Program (USDA/PDP). Imidacloprid and thiamethoxam were the most commonly detected neonicotinoids in fruits and vegetables with 66 and 51% detection in the HZC study and 52 and 53% detection in the USCC study, respectively. The overall frequency of detection for neonicotinoids in the USDA/PDP samples was much lower than those reported here for the USCC or HZC studies, with imidacloprid being the most frequently detected neonicotinoid at 7.3%. The high frequencies of neonicotinoid detection in fruits and vegetables in the USCC and HZC studies give us a snapshot of the ubiquity of neonicotinoid use in global agriculture and make it clear that neonicotinoids have become part of the dietary staple, with possible health implications for individuals.


Physiological responses to divergent selection for phytate phosphorus bioavailability in a randombred chicken population.
An investigation was conducted to study insulin-like growth factor (IGF)-I, IGF-II, insulin, glucagon, leptin, triiodothyronine (T(3)), and thyroxine (T(4)) levels in a chicken population divergently selected for P bioavailability (PBA). There were differences in growth and feed efficiency between the 2 lines. Concentrations of IGF-I, IGF-II, and T(3) were significantly greater in the high PBA line compared with the low PBA line, whereas the reverse was true for glucagon. There were no correlations between IGF-I and II and PBA in either line, suggesting that the line differences may be the result of factors other than PBA. Glucagon and IGF-I have different relationships with feed conversion ratio in the high PBA line compared with the low PBA line. There was a significant correlation between PBA and T(3) in the low line and between PBA and T(4) in the high PBA line. Thyroid hormone levels may be an indirect indicator of PBA in growing chickens. The genes in the thyroid hormone pathway may be key in the identification of genes associated with PBA.


Treatment of osteochondral defects with autologous bone marrow in a hyaluronan-based delivery vehicle.
The natural repair of osteochondral defects can be enhanced with biocompatible, biodegradable and bioactive materials that provide structural support and molecular cuing to stimulate repair. Since bone marrow contains osteochondral progenitor cells and bioactive agents, it is hypothesized that the combination of scaffold and bone marrow would be a superior composite material for osteochondral repair. This hypothesis will be tested by comparing the outcome of osteochondral defects filled with a fibronectin-coated hyaluronan-based sponge (ACP) with or without autologous bone marrow. Thirty-three 4-month-old rabbits received 3-mm diameter osteochondral defects that were then filled with ACP loaded or not with autologous bone marrow. Rabbits were sacrificed at 2, 3, 4, 12, and 24 weeks after surgery and the condyles processed for histologic and immunohistochemical evaluation. The defects were graded with a histologic scoring scale. Except for the 3-week specimens, the histologic appearance of the defects was similar in both groups. Four weeks after surgery, the defects were filled with bone with a top layer of cartilage well integrated with the adjacent cartilage. Twelve and 24 weeks after surgery, the defects again showed bone filling. The primary difference between the 4-week samples and the 12- and 24-week samples was that the layer of cartilage that appeared to be thinner than the adjacent cartilage. At each harvest time, the overall histologic scores of the specimens did not reveal statistical differences between the treatment groups. However, as revealed by the results of the 3-week sacrifices, bone marrow loading appeared to accelerate the first stages of the repair process. The fibronectin-coated hyaluronan-based scaffold appears to organize the natural response and facilitate the integration of the neo-cartilage with the adjacent tissue. The fundamental tissue engineering principles derived from this study should provide guidelines for the development of comparable clinical reconstructive therapies.


Isolation and characterization of the mouse CD8 beta-chain (Ly-3) genes. Absence of an intervening sequence between V- and J-like gene segments.
We have isolated and determined the nucleotide sequence and genomic organization of the genes encoding Ly-3.1 and Ly-3.2. These genes span approximately 14 kb on chromosome 6 and consist of six exons and five introns. The exons correlate roughly with the putative functional domains, namely, a leader exon, a variable and joining region-like exon, a hinge region-like exon, a transmembrane exon, and two intracytoplasmic exons. There is no intervening sequence between V- and J-like gene segments, indicating that rearrangement is not necessary for the expression of the Ly-3 gene. In the 5'-flanking region there is no "TATA" box nor "CAAT" box; however, three "GC" boxes are located upstream of the ATG initiator codon. There are short stretches of sequence homologous to 5'-flanking sequences of the Ly-2 gene. In addition, the sequences CTCTGTGGCA at -748 exhibits homology to the enhancer core sequence of the human Ig H chain and TCR genes. Comparison of the nucleotide sequence corresponding to the extracellular portion between Ly-3.1 and Ly-3.2 revealed a single base difference which results in an amino acid substitution. Therefore it is likely that this amino acid difference is responsible for the previously defined Ly-3 allotypes.


Expression of hypoxia-inducible genes is suppressed in altered gravity due to impaired nuclear HIF1α accumulation.
Extravehicular activities, the backbone of manned space exploration programs, set astronauts into mild hypoxia. Unfortunately, microgravity aggravates threatening symptoms of hypoxia such as vision impairment and brain edema. Hypoxia-inducible factors (HIFs) sense cellular hypoxia and, subsequently, change the cells' expression profile instantaneously by rapidly translocating-most likely cytoskeleton-dependently-into the nucleus and subsequently forming transcription complexes with other proteins. We tested the hypothesis that this fundamental process could be altered by sudden changes in gravitational forces in parabolic flights using a newly developed pocket-size cell culture lab that deoxygenizes cells within 15 min. Sudden gravity changes (SGCs 1g-1.8g-0g-1.8g-1g) during hypoxic exposure suppressed expression of the HIF1α-dependent genes investigated as compared with hypoxia at constant 1g. Normoxic cells subjected to SGCs showed reduced nuclear but not cytoplasmatic HIF1α signal and appeared to have disturbed cytoskeleton architecture. Inhibition of the actin-dependent intracellular transport using a combination of myosin V and VI inhibitors during hypoxia mimicked the suppression of the HIF1α-dependent genes observed during hypoxic exposure during SGCs. Thus, SGCs seem to disrupt the cellular response to hypoxia by impairing the actin-dependent translocation of HIF1α into the nucleus.


Peak expiratory flow rate of children working in lock factories.
Peak expiratory flow rate (PEFR) of 106 children working in different units of lock factory was measured and compared with age and sex matched control group of same socio-economic status children. All the children worked for about ten hours per day. It was observed that there was a significant decrease in PEFR of children working in the different units of lock factories i.e. Hand press, Polishing, Lock fitting, Lock packing units as compared to control group (P>0.001). The reduction percentage of PEFR was maximum in children working in polishing unit (25.48%).


Cytochrome P450 metabolizing fatty acids in plants: characterization and physiological roles.
In plants, fatty acids (FA) are subjected to various types of oxygenation reactions. Products include hydroxyacids, as well as hydroperoxides, epoxides, aldehydes, ketones and α,ω-diacids. Many of these reactions are catalysed by cytochrome P450s (P450s), which represent one of the largest superfamilies of proteins in plants. The existence of P450-type metabolizing FA enzymes in plants was established approximately four decades ago in studies on the biosynthesis of lipid polyesters. Biochemical investigations have highlighted two major characteristics of P450s acting on FAs: (a) they can be inhibited by FA analogues carrying an acetylenic function, and (b) they can be enhanced by biotic and abiotic stress at the transcriptional level. Based on these properties, P450s capable of producing oxidized FA have been identified and characterized from various plant species. Until recently, the vast majority of characterized P450s acting on FAs belonged to the CYP86 and CYP94 families. In the past five years, rapid progress in the characterization of mutants in the model plant Arabidopsis thaliana has allowed the identification of such enzymes in many other P450 families (i.e. CYP703, CYP704, CYP709, CYP77, CYP74). The presence in a single species of distinct enzymes characterized by their own regulation and catalytic properties raised the question of their physiological meaning. Functional studies in A. thaliana have demonstrated the involvement of FA hydroxylases in the synthesis of the protective biopolymers cutin, suberin and sporopollenin. In addition, several lines of evidence discussed in this minireview are consistent with P450s metabolizing FAs in many aspects of plant biology, such as defence against pathogens and herbivores, development, catabolism or reproduction.


[Rapid detection of genotypes of TT virus using a heteroduplex mobility assay].
To establish a simple, sensitive, specific and less-costly method for detecting genotypes of TT virus (TTV). TTV DNA was tested by nested polymerase chain reaction (nPCR) in sera from 180 patients with different types of viral hepatitis and 96 normal individuals in Beijing. TTV genotypes were determined in 40 sera collected from TTV DNA positive patients by heteroduplex mobility assay (HMA) and through sequencing. The positive rates of TTV DNA in viral hepatitis patients and normal individuals were 22.2% (40/180) and 19.8% (19/96), respectively (chi(2) = 0.220, P = 0.639). TTV DNA positive rates of patients with hepatitis A, B, C, E and non-A to E were 20.0% (6/30), 16.7% (5/30), 23.3% (7/30), 36.7% (11/30) and 18.3% (11/60), respectively. Of 40 TTV DNA positive patients, 20 (50.0%) were TTV G1, 7 (17.5%) TTV G2, 10 (25.0%) coinfected with different genotypes of TTV, and 3 untyped by HMA. Twenty G1 and 7 G2 detected by HMA were confirmed by sequence analysis. Of 10 patients coinfected with different genotypes of TTV, 5 were G1 and G2, 2 G1 and G3, 1 G1 and G4, 1 G1 and G3, and 1 with G1, G2 and G3 coinfections. HMA was recognized as simple, sensitive, specific and less-costly, thus could be used for genotyping of TTV.