Metagenome reveals potential microbial degradation of hydrocarbon coupled with sulfate reduction in an oil-immersed chimney from Guaymas Basin.
Deep-sea hydrothermal vent chimneys contain a high diversity of microorganisms, yet the metabolic activity and the ecological functions of the microbial communities remain largely unexplored. In this study, a metagenomic approach was applied to characterize the metabolic potential in a Guaymas hydrothermal vent chimney and to conduct comparative genomic analysis among a variety of environments with sequenced metagenomes. Complete clustering of functional gene categories with a comparative metagenomic approach showed that this Guaymas chimney metagenome was clustered most closely with a chimney metagenome from Juan de Fuca. All chimney samples were enriched with genes involved in recombination and repair, chemotaxis and flagellar assembly, highlighting their roles in coping with the fluctuating extreme deep-sea environments. A high proportion of transposases was observed in all the metagenomes from deep-sea chimneys, supporting the previous hypothesis that horizontal gene transfer may be common in the deep-sea vent chimney biosphere. In the Guaymas chimney metagenome, thermophilic sulfate reducing microorganisms including bacteria and archaea were found predominant, and genes coding for the degradation of refractory organic compounds such as cellulose, lipid, pullullan, as well as a few hydrocarbons including toluene, ethylbenzene and o-xylene were identified. Therefore, this oil-immersed chimney supported a thermophilic microbial community capable of oxidizing a range of hydrocarbons that served as electron donors for sulphate reduction under anaerobic conditions.


Activation of the PAK-related kinase by human immunodeficiency virus type 1 Nef in primary human peripheral blood lymphocytes and macrophages leads to phosphorylation of a PIX-p95 complex.
Human immunodeficiency virus type 1 (HIV-1) Nef enhances virus replication in both primary T lymphocytes and monocyte-derived macrophages. This enhancement phenotype has been linked to the ability of Nef to modulate the activity of cellular kinases. We find that despite the reported high-affinity interaction between Nef and the Src kinase Hck in vitro, a Nef-Hck interaction in the context of HIV-1-infected primary macrophages is not detectable. However, Nef binding and activation of the PAK-related kinase and phosphorylation of its substrate could be readily detected in both infected primary T lymphocytes and macrophages. Furthermore, we show that this substrate is a complex composed of the recently characterized PAK interacting partner PIX (PAK-interacting guanine nucleotide exchange factor) and its tightly associated p95 protein. PAK and PIX-p95 appear to be differentially activated and phosphorylated depending on the intracellular environment in which nef is expressed. These results identify the PIX-p95 complex as a novel effector of Nef in primary cells and suggest that the regulation of the PAK signaling pathway may differ in T cells and macrophages.


[Characteristics of phenotypes of Alteromonas putrefaciens. Study of 123 strains].
The phenotypic characteristics of 124 strains of Alteromonas putrefaciens, a bacterium that produces large quantities of hydrogen sulfide, were studied. The strains were isolated from man, warm and cold-blooded animals (especially fish and shellfish), marine and fresh waters. Two phenetic groups were differentiated: group A strains grew at 42 degrees C, but not at 4 degrees C, tolerated 6 p. cent NaCl and did not utilize glucose or maltose, unlike group B strains. Each group could be divided into two subgroups A1, A2 and B1, B2. Citrate utilization, hydrolysis of urea and aesculin, resistance or susceptibility to cephalothin and vibriostatic agent 0/129 were useful for subgroup determination. The taxonomic relatedness of A. putrefaciens to genus Alteromonas are discussed.


The penetration of antibiotics into bacterial cells: significance to antimicrobial action.
The penetration of antibiotics into the microbial cell is reviewed from the standpoint of the structure and function of the several types of membranes which constitute permeability barriers to the passage of hydrophilic molecules. It is clear that little is actually known about the mechanisms by which the majority of antibiotics gain access to their site of action within the target cell. Sufficient evidece is at hand to indicate that resistance to several groups of antibiotics is determined by changes in the permeability of the cell membranes, the genes for which are often carried on R-plasmids. The importance of studies to determine the precise mechanisms by which each type of antibiotic permeates the membrane of the bacterial cell is stressed by a discussion of several approaches to enhancing antibiotic permeation through manipulation of either the drug or the target cell based on knowledge of the uptake mechanisms.


The distribution of intraocular pressure and its association with metabolic syndrome in a community.
The current study was performed to assess the distribution of intraocular pressure (IOP) and its association with metabolic syndrome (MS) in a community. We measured IOP and MS components from 446 adults, age 20 or more years old, who reside in a community in Kyunggi Province, South Korea. We compared the level of IOP according to the number of metabolic abnormalities and between normal and abnormal metabolic components. Linear regression analyses were used to determine the relationship between IOP and metabolic components. No significant difference in IOP (mean +/- SE) was found between men (12.24 +/- 2.42) and women (12.55 +/- 2.41 mmHg, p > 0.1), while IOP of men tended to decrease as age increased (p for trend < 0.01). After adjusting for age, IOP of subjects with abdominal obesity in men and high blood pressure in women were significantly higher than those without abdominal obesity or high blood pressure (p < 0.05). Female subjects with MS showed significantly higher IOP than those without MS. Participants with more metabolic disturbances tended to have a greater IOP elevation with a linear trend after adjusting for age and sex. In the univariate regression analysis, age and waist circumference were significantly associated with IOP in men, but systolic and diastolic blood pressure were associated with IOP in women. In final multiple regression model, age, systolic blood pressure, and triglyceride were associated with IOP in women, and age in men. These findings suggest that MS and its components may be important determinants of elevated IOP.


Titania-promoted carboxylic acid alkylations of alkenes and cascade addition-cyclizations.
Photochemical reactions employing TiO2 and carboxylic acids under dry anaerobic conditions led to several types of C-C bond-forming processes with electron-deficient alkenes. The efficiency of alkylation varied appreciably with substituents in the carboxylic acids. The reactions of aryloxyacetic acids with maleimides resulted in a cascade process in which a pyrrolochromene derivative accompanied the alkylated succinimide. The selectivity for one or other of these products could be tuned to some extent by employing the photoredox catalyst under different conditions. Aryloxyacetic acids adapted for intramolecular ring closures by inclusion of 2-alkenyl, 2-aryl, or 2-oximinyl functionality reacted rather poorly. Profiles of reactant consumption and product formation for these systems were obtained by an in situ NMR monitoring technique. An array of different catalyst forms were tested for efficiency and ease of use. The proposed mechanism, involving hole capture at the TiO2 surface by the carboxylates followed by CO2 loss, was supported by EPR spectroscopic evidence of the intermediates. Deuterium labeling indicated that the titania likely donates protons from surface hydroxyl groups as well as supplying electrons and holes, thus acting as both a catalyst and a reaction partner.


Cardiac and renal protective effects of 2,5-dimethylcelecoxib in angiotensin II and high-salt-induced hypertension model mice.
We reported that 2,5-dimethylcelecoxib (DM-celecoxib), a celecoxib derivative that is unable to inhibit cyclooxygenase-2, prevented cardiac remodeling induced by sarcomeric gene mutation, left ventricular pressure overload, or β-adrenergic receptor stimulation. This effect seemed to be mediated by the inhibition of the canonical Wnt/β-catenin signaling pathway, which has been suggested to play a key role in the development of chronic kidney disease and chronic heart failure. We investigated the effect of DM-celecoxib on cardiac remodeling and kidney injury in hypertension model mice induced by angiotensin II infusion in the absence or presence of high-salt load. DM-celecoxib prevented cardiac remodeling and markedly reduced urinary albumin excretion without altering blood pressure in those mice. Moreover, DM-celecoxib prevented podocyte injury, glomerulosclerosis, and interstitial fibrosis in the kidney of mice loaded with angiotensin II and high-salt load. DM-celecoxib reduced the phosphorylation level of Akt and activated glycogen synthase kinase-3, which led to the suppression of the Wnt/β-catenin signal in the heart and kidney. DM-celecoxib also reduced the expression level of snail, a key transcription factor for the epithelial-mesenchymal transition and of which gene is a target of the Wnt/β-catenin signal. Results of the current study suggested that DM-celecoxib could be beneficial for patients with hypertensive heart and kidney diseases.


[Evaluation of effectiveness of short-term orthostatic positioning of head-down tilted primates on the liquid body sectors].
Head-down tilting (HDT) of primates is a well-recognized model for studying the hypokinetic syndrome effects on the body functions. Benefits for the body liquids distribution of periodic transition of primates (rhesus macaques) into the vertical position without support loading were evaluated in a 30-d HDT experiment. Brief (30 to 120 minutes long, 4-5 times a week) transition from tilting into the vertical position had no influence on primates' hydration homeostasis and its infrastructure. However, the orthostatic test at the end of the HDT experiment showed a less increase in blood content of the lower extremities as compared with tilted primates that had not been periodically turned up into the vertical position.


Renin angiotensin aldosterone system blockade and renal disease in patients with type 2 diabetes. An Asian perspective from the RENAAL Study.
Asia is predicted to have the largest population of patients with diabetes who are at high risk for renal disease. In the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study, approximately 17% of patients were Asians. In this subgroup analysis, we examined the characteristics, response, and adherence to treatment of the Asian population, as well as their baseline predictors of risk of renal end points. A total of 252 Asian patients were enrolled in the RENAAL study, which compared losartan (50 mg titrated to 100 mg) to placebo in addition to conventional antihypertensive medications in type 2 diabetic patients with nephropathy. Mean follow-up was 3.2 years. The effect of losartan therapy on renal and cardiovascular outcomes was examined, and baseline predictors of risk were determined using a Cox proportional hazards model with prespecified baseline covariates. Losartan reduced the risk of the primary composite end point composed of a doubling of serum creatinine, end-stage renal disease, or all-cause mortality in Asian patients by 35% (P = 0.02). No difference between losartan and placebo was observed for the cardiovascular composite outcomes. Losartan reduced the level of proteinuria by 47% (P < 0.001) and rate of decrease in renal function by 31% (0.0074). Discontinuations were lower in the losartan-treated patients. The strongest baseline predictors of risk of renal end points were proteinuria (hazard ratio 1.42, P < 0.0001) and low Hb (0.81, P < 0.0001). In this subgroup analysis of the RENAAL study, losartan conferred significant renal benefits and was well tolerated in Asian patients with type 2 diabetes and clinical nephropathy. Baseline proteinuria and low Hb were strong predictors of risk of renal outcomes.


Nitric oxide, human diseases and the herbal products that affect the nitric oxide signalling pathway.
1. Nitric oxide (NO) is formed enzymatically from l-arginine in the presence of nitric oxide synthase (NOS). Nitric oxide is generated constitutively in endothelial cells via sheer stress and blood-borne substances. Nitric oxide is also generated constitutively in neuronal cells and serves as a neurotransmitter and neuromodulator in non-adrenergic, non-cholinergic nerve endings. Furthermore, NO can also be formed via enzyme induction in many tissues in the presence of cytokines. 2. The ubiquitous presence of NO in the living body suggests that NO plays an important role in the maintenance of health. Being a free radical with vasodilatory properties, NO exerts dual effects on tissues and cells in various biological systems. At low concentrations, NO can dilate the blood vessels and improve the circulation, but at high concentrations it can cause circulatory shock and induce cell death. Thus, diseases can arise in the presence of the extreme ends of the physiological concentrations of NO. 3. The NO signalling pathway has, in recent years, become a target for new drug development. The high level of flavonoids, catechins, tannins and other polyphenolic compounds present in vegetables, fruits, soy, tea and even red wine (from grapes) is believed to contribute to their beneficial health effects. Some of these compounds induce NO formation from the endothelial cells to improve circulation and some suppress the induction of inducible NOS in inflammation and infection. 4. Many botanical medicinal herbs and drugs derived from these herbs have been shown to have effects on the NO signalling pathway. For example, the saponins from ginseng, ginsenosides, have been shown to relax blood vessels (probably contributing to the antifatigue and blood pressure-lowering effects of ginseng) and corpus cavernosum (thus, for the treatment of men suffering from erectile dysfunction; however, the legendary aphrodisiac effect of ginseng may be an overstatement). Many plant extracts or purified drugs derived from Chinese medicinal herbs with proposed actions on NO pathways are also reviewed.


Aseismic uplift in southern california.
Preliminary examination of the historic geodetic record has disclosed crustal uplift of 0.15 to 0.25 meter that apparently began around 1960 and has since grown to include at least 12,000 square kilometers of southern California. This uplift extends at least 150 kilometers west-northwestward along the San Andreas Fault from Cajon to Maricopa, southward from the San Andreas into the northern Transverse Ranges, and eastward from Lebec into and including much of western Mojave block. It seems to have grown spasmodically eastward from a center near the junction of the San Andreas and Garlock faults and has occurred largely within an area that has remained virtually aseismic since at least 1932. Although much of this area has been characterized by crustal mobility since at least the turn the century, the described uplift seems to be an unusually large and probably unique event superimposed the existing pattern of continuing deformation.


Assessing Age Inclusivity in Higher Education: Introducing the Age-Friendly Inventory and Campus Climate Survey.
This study introduces a theoretical framework for assessing age inclusivity in higher education environments and describes the Age-Friendly Inventory and Campus Climate Survey (ICCS). The ICCS measures age-friendly campus practices as reported by administrators, perceptions of age-friendliness by campus constituents, and the fit between objective practices and subjective perceptions as an overall indicator of age inclusivity. The ICCS was administered at a public university in the northeastern United States. Administrators completed the Inventory of potential age-friendly campus practices associated with their units. Campus constituents (n = 688) completed the online Campus Climate Survey to assess subjective awareness of these practices, perceived age-friendliness, and personal beliefs about age inclusivity. The Inventory yielded a score of 66% of potential age-friendly practices in place as reported by administrators. The Campus Climate Survey showed low overall perceptions of age-friendliness and varied beliefs about age inclusivity on campus. Fit was measured by comparing the Inventory practices and Campus Climate Survey awareness of the existence of 47 of 73 potential practices. Convergence on this campus showed an awareness of 36% of age-friendly practices. Based on the proposed theoretical framework for age inclusivity, the ICCS offers a way of assessing the age-friendliness of the objective environment across campus functions, the subjective environment across campus constituents, and the fit between subjective and objective environments on campuses. The ICCS will help higher education institutions identify strengths and challenges for advancing age inclusivity.


Robust voltage-controlled transcutaneous energy transfer system for artificial anal sphincter.
The artificial anal sphincter (AAS) system has gained significant attention as a solution for treating fecal incontinence (FI). It relies on transcutaneous energy transfer (TET) as its primary energy source. However, changes in posture or biological tissue can cause misalignment of the coil, resulting in unstable power reception. Inadequate power affects charging efficiency, while excessive power leads to excessive heating at the receiver side. Consequently, achieving safe and constant voltage charging for the AAS becomes a complex challenge. To maintain a consistent charging voltage and overcome the issue of variations in load and coil coupling strength, this article proposes a wireless charging control system that utilizes an LCC-S-type resonant network and phase shift to adjust the transmitting voltage based on feedback charging voltage in real time. In particular, the PI controller and neural network are introduced to change the phase-shift angle swiftly. The dynamic performance is then evaluated under different misalignments and presented with comparative results. The results indicate that the multilayer perceptron control system outperforms the PI. Under the complex misalignment disturbance, the average error of receiver side load voltage is only 0.007 V, with an average settling time of 960 ms. Additionally, the average temperature at the receiver side is 40.4°C. The experiments demonstrate that the proposed system effectively addresses the misalignment issue in TET during the charging, ensuring constant voltage charging at the receiver side and thermal safety.


Solid leukemic tumor. An uncommon presentation of a common disease.
Granulocytic sarcoma of the clavicle occurred in a 17-year-old boy. The patient had a densely sclerotic bony lesion with periosteal reaction and no peripheral blood manifestations of leukemia. Supraclavicular adenopathy developed after a preliminary diagnosis of Ewing's sarcoma was made, and a second biopsy was performed. Electron microscopy of the tissue showed Auer bodies and Charcot-Leyden granules characteristic of acute myelogenous leukemia. Combination chemotherapy, local irradiation, and clavicular resection have left the patient with no evidence of disease 50 months after diagnosis. Leukemia may appear as a solitary bone tumor before hematologic changes develop. Granulocytic sarcomas can have the radiographic and histologic appearance of primary bone tumors. Bone marrow examination and electron microscopy of biopsy tissue are diagnostic of leukemia and are important in all hospital investigations of round-cell tumors of bone.


Hearing Aid Noise Reduction Lowers the Sustained Listening Effort During Continuous Speech in Noise-A Combined Pupillometry and EEG Study.
The investigation of auditory cognitive processes recently moved from strictly controlled, trial-based paradigms toward the presentation of continuous speech. This also allows the investigation of listening effort on larger time scales (i.e., sustained listening effort). Here, we investigated the modulation of sustained listening effort by a noise reduction algorithm as applied in hearing aids in a listening scenario with noisy continuous speech. The investigated directional noise reduction algorithm mainly suppresses noise from the background. We recorded the pupil size and the EEG in 22 participants with hearing loss who listened to audio news clips in the presence of background multi-talker babble noise. We estimated how noise reduction (off, on) and signal-to-noise ratio (SNR; +3 dB, +8 dB) affect pupil size and the power in the parietal EEG alpha band (i.e., parietal alpha power) as well as the behavioral performance. Our results show that noise reduction reduces pupil size, while there was no significant effect of the SNR. It is important to note that we found interactions of SNR and noise reduction, which suggested that noise reduction reduces pupil size predominantly under the lower SNR. Parietal alpha power showed a similar yet nonsignificant pattern, with increased power under easier conditions. In line with the participants' reports that one of the two presented talkers was more intelligible, we found a reduced pupil size, increased parietal alpha power, and better performance when people listened to the more intelligible talker. We show that the modulation of sustained listening effort (e.g., by hearing aid noise reduction) as indicated by pupil size and parietal alpha power can be studied under more ecologically valid conditions. Mainly concluded from pupil size, we demonstrate that hearing aid noise reduction lowers sustained listening effort. Our study approximates to real-world listening scenarios and evaluates the benefit of the signal processing as can be found in a modern hearing aid.


Reconstruction and analysis of exploding wire particle trajectories via automatic calibration of stereo images.
Quantitative understanding of the physics of dust or granular matter transport significantly impacts several aspects of burning plasma science and technology. This work takes machine vision techniques popular in robotics and self-driving cars and applies them to identification and analysis of microparticles generated from exploding wires. Using only the image frames and knowledge of the intrinsic properties of the cameras, a Python code was written to identify the particles, automatically calibrate the relative image positions, and extract trajectory data. After identifying approximately 50 particles based on the timing of secondary particle explosions, the eight point and random sample consensus algorithms were used to determine the geometric correlation between the cameras. Over 100 particle matches were found between the two camera views. These correlated trajectories were used in subsequent 3D track reconstruction and analysis of the physics behind the particle motion. The 3D reconstruction resulted in accurate positioning of the particles with respect to the experimental setup. The particle motion was consistent with the effects of a 1 g gravitational field modified by drag forces. The methods and analyses presented here can be used in many facets of high temperature plasma diagnostics.


Sexual risk and sexual healthcare utilization profiles among Black sexual minority men in the U.S. Deep South.
Black gay, bisexual, and other Black sexual minority men (BSMM) continue to experience negative health outcomes along the HIV prevention and treatment continuum, especially in the U.S. Deep South. The purpose of this study is to identify sexual risk and healthcare utilization profiles behaviors among BSMM in the Deep South. Guided by the Behavioral Model for Vulnerable Populations, latent class analysis was used to identify sexual risk and healthcare utilization profiles using data from 348 individuals in Jackson, MS and Atlanta, GA. Multinomial logistic regression was used to identify the correlates of class membership. A 3-class solution was identified: Class 1 (Substance using, Low Testers); Class 2 (Condom using testers), and Class 3 (Casual Partner Testers). Class 1 had the highest conditional probabilities of drug (75%) and alcohol (84%) use before sex and the lowest probability of STI and HIV testing. Class 2 had the highest probabilities of condom use and a 65% probability of seeing a healthcare provider. Class 3 had the highest probability of inconsistent condom use and seeing a healthcare provider in the previous 12 months (76%). Findings support the need for targeted interventions tailored to BSMM of different sexual risk and healthcare utilization behaviors.


Image Segmentation Using Parametric Contours With Free Endpoints.
In this paper, we introduce a novel approach for active contours with free endpoints. A scheme for image segmentation is presented based on a discrete version of the Mumford-Shah functional where the contours can be both closed and open curves. Additional to a flow of the curves in normal direction, evolution laws for the tangential flow of the endpoints are derived. Using a parametric approach to describe the evolving contours together with an edge-preserving denoising, we obtain a fast method for image segmentation and restoration. The analytical and numerical schemes are presented followed by numerical experiments with artificial test images and with a real medical image.


Computer-assisted measurement of primary tumor area is prognostic of recurrence-free survival in stage IB melanoma patients.
Current staging guidelines are insufficient to predict which patients with thin primary melanoma are at high risk of recurrence. Computer-assisted image analysis may allow for more practical and objective histopathological analysis of primary tumors than traditional light microscopy. We studied a prospective cohort of stage IB melanoma patients treated at NYU Langone Medical Center from 2002 to 2014. Primary tumor width, manual area, digital area, and conformation were evaluated in a patient subset via computer-assisted image analysis. The associations between histologic variables and survival were evaluated using Cox proportional hazards model. Logistic regressions were used to build a classifier with clinicopathological characteristics to predict recurrence status. Of the 655 patients with stage IB melanoma studied, a subset of 149 patient tumors (63 recurred, 86 did not recur) underwent computer-assisted histopathological analysis. Increasing tumor width (hazard ratios (HR): 1.17, P=0.01) and digital area (HR: 1.08, P<0.01) were significantly associated with worse recurrence-free survival, whereas non-contiguous conformation (HR: 0.57, P=0.05) was significantly associated with better recurrence-free survival. The novel histopathological classifier composed of digital area, conformation, and baseline variables effectively distinguished recurrent cases from non-recurrent cases (AUC: 0.733, 95% confidence interval (CI): 0.647-0.818), compared to the baseline classifier alone (AUC: 0.635, 95% CI: 0.545-0.724). Primary tumor cross-sectional area, width, and conformation measured via computer-assisted analysis may help identify high-risk patients with stage IB melanoma.


Chemical approaches for functionally probing the proteome.
With the availability of complete genome sequences, emphasis has shifted toward the understanding of protein function. We have developed a functional proteomic methodology that makes use of chemically reactive fluorescent probes to profile and identify enzymes in complex mixtures by virtue of their catalytic activity. This methodology allows a comparison of changes in activity of multiple enzymes under a variety of conditions using a single two-dimensional separation. The probes can also be used to localize active enzymes in intact cells using fluorescence microscopy. Furthermore, the probes enable screens for selective small molecule inhibitors of each enzyme family member within crude lysates or intact cells. Ultimately, this technology allows the rapid identification of potential drug targets and small molecule lead compounds targeted to them.


Lysates of turkey-grown Pasteurella multocida: effects of solubilizing agents on the immunologic properties of membrane vesicles.
Membrane vesicles from lysed suspensions of turkey-grown Pasteurella multocida were treated with various solubilizing agents to release protein that may contain cross-protection factor. Potassium thiocyanate, NaOH-glycine, lithium diiodosalicylate, guanidine hydrochloride, n-butanol, dimethyl sulfoxide, Triton X-100, and sodium lauryl sarcosinate were each tested as solubilizing agents. Vaccines made from combining solubilized membrane vesicles with complete lysate supernatant fluid produced various degrees of protection against challenge exposure with a heterologous serotype of P multocida in turkeys. Only vaccines prepared from membranes that were solubilized with potassium thiocyanate and sodium lauryl sarcosinate protected as well as complete lysate from turkey-grown P multocida. The amount of protein in each vaccine did not relate to protection. Distinct chemical differences were observed between lysates prepared from turkey-grown P multocida and lysates prepared from 41 C broth-grown P multocida. The external morphology of P multocida, after treatment with lysozyme and EDTA, was similar whether grown in broth or in turkeys.


Dichotic listening asymmetry: sex differences and menstrual cycle effects.
The impact of menstrual cyclicity and sex differences on dichotic listening was studied in 25 women and 20 men (aged 20-25 years). Dichotic listening was administered using consonant-vowel (CV) stimuli and tested across three attention conditions. Women were tested at two points in the menstrual cycle (Day 2-5: low oestrogen and progesterone/Low-EP; Day 18-25: high oestrogen and progesterone/High-EP). Men were tested once. Performance averaged across attention conditions was analysed for menstrual cycle and sex effects. Significant menstrual cycle phase effects were observed in women. At the High-EP phase women showed a greater right ear advantage (REA) compared to the Low-EP phase. Sex differences were found when dichotic listening asymmetry in men was compared to women at the Low-EP, but not the High-EP phase. In contrast to laterality effects, baseline perceptual performance (total right plus left ear response) was similar in men and women at both phases of the menstrual cycle. Results support a role for ovarian hormones in shaping laterality of speech perception in women. This study also emphasises the importance of considering menstrual cycle effects when evaluating sex differences in dichotic listening.


Cochlear abnormalities in Alport's syndrome.
The cochleae of four patients with a diagnosis or suspicion of Alport's syndrome were examined by microdissection and surface preparations. Strial atrophy, found in all cases, was most pronounced in the middle and apical turns and mainly involved the inferior margin of the stria. Numerous vacuoles were seen in the spiral ligament behind the atrophic stria. The youngest patient, aged 16 years, who had little or no hearing loss, showed limited hair cell degeneration. The other two patients had severe hearing loss and extensive sensory cell loss of unusual distribution. In the specimens from the second patient, the extent of hair cell loss was fairly even throughout, but the loss of inner hair cells was far more severe than the loss of outer hair cells. In the third case, the loss of outer hair cells was most pronounced in the apical half of the cochlea. All of these specimens had only a mild secondary nerve degeneration. The specimens from the fourth patient were poorly preserved and showed only nerve degeneration.


Accuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymerase.
For certain applications of the polymerase chain reaction (PCR), it may be necessary to consider the accuracy of replication. The breakthrough that made PCR user friendly was the commercialization of Thermus aquaticus (Taq) DNA polymerase, an enzyme that would survive the high temperatures needed for DNA denaturation. The development of enzymes with an inherent 3' to 5' exonuclease proofreading activity, lacking in Taq polymerase, would be an improvement when higher fidelity is needed. We used the forward mutation assay to compare the fidelity of Taq polymerase and Thermotoga maritima (ULTMA) DNA polymerase, an enzyme that does have proofreading activity. We did not find significant differences in the fidelity of either enzyme, even when using optimal buffer conditions, thermal cycling parameters, and number of cycles (0.2% and 0.13% error rates for ULTMA and Taq, respectively, after reading about 3,000 bases each). We conclude that for sequencing purposes there is no difference in using a DNA polymerase that contains an inherent 3' to 5' exonuclease activity for DNA amplification. Perhaps the specificity and fidelity of PCR are complex issues influenced by the nature of the target sequence, as well as by each PCR component.


Preparation and characterization of transparent hydrophilic photocatalytic TiO2/SiO2 thin films on polycarbonate.
Transparent hydrophilic photocatalytic TiO2 coatings have been widely applied to endow the surfaces self-cleaning properties. A mixed metal oxide (TiO2/SiO2) can enhance the photocatalytic performance improving the ability of surface adsorption and increasing the amount of hydroxyl surface groups. The present work introduces a systematic study concerning the effect of the SiO2 addition to TiO2 films on the wettability, the photocatalytic activity, the adhesion strength, and the mechanical stability of the films. Transparent hydrophilic photocatalytic TiO2/SiO2 thin films were used to coat the polycarbonate (PC) substrate which was precoated by an intermediate SiO2 layer. The TiO2/SiO2 thin film was prepared employing a bulk TiO2 powder (Sachtleben Hombikat UV 100) and different molar ratios of tetraethoxysilane in acidic ethanol. A dip-coating process was used to deposit the films onto the polycarbonate substrate. The films were characterized by UV/vis spectrophotometry, FTIR spectroscopy, ellipsometry, BET, AFM, XRD, and water contact angle measurements. The mechanical stability and the UV resistance were examined. The photocatalytic activity of the coated surface was calculated from the kinetic analysis of methylene blue photodegradation measurements and compared with the photocatalytic activity of Pilkington Activ sheet glass. The coated surfaces displayed considerable photocatalytic activity and superhydrophilicity after exposure to UV light. The addition of SiO2 results in an improvement of the photocatalytic activity of the TiO2 film reaching the highest value at molar ratio TiO2/SiO2 equal to 1:0.9. The prepared films exhibit a good stability against UV(A) irradiation.


Entomological risk assessment for transmission of arboviral diseases by Aedes mosquitoes in a domestic and forest site in Accra, Ghana.
Dengue, Zika and chikungunya are Aedes-borne viral diseases that have become great global health concerns in the past years. Several countries in Africa have reported outbreaks of these diseases and despite Ghana sharing borders with some of these countries, such outbreaks are yet to be detected. Viral RNA and antibodies against dengue serotype-2 have recently been reported among individuals in some localities in the regional capital of Ghana. This is an indication of a possible silent transmission ongoing in the population. This study, therefore, investigated the entomological transmission risk of dengue, Zika and chikungunya viruses in a forest and domestic population in the Greater Accra Region, Ghana. All stages of the Aedes mosquito (egg, larvae, pupae and adults) were collected around homes and in the forest area for estimation of risk indices. All eggs were hatched and reared to larvae or adults for morphological identification together with larvae and adults collected from the field. The forest population had higher species richness with 7 Aedes species. The predominant species of Aedes mosquitoes identified from both sites was Aedes aegypti (98%). Aedes albopictus, an important arbovirus vector, was identified only in the peri-domestic population at a prevalence of 1.5%, significantly higher than previously reported. All risk indices were above the WHO threshold except the House Index for the domestic site which was moderate (19.8). The forest population recorded higher Positive Ovitrap (34.2% vs 26.6%) and Container (67.9% vs 36.8%) Indices than the peri-domestic population. Although none of the mosquito pools showed the presence of dengue, chikungunya or Zika viruses, all entomological risk indicators showed that both sites had a high potential arboviral disease transmission risk should any of these viruses be introduced. Continuous surveillance is recommended in these and other sites in the Metropolis to properly map transmission risk areas to inform outbreak preparedness strategies.


A trivariate continual reassessment method for phase I/II trials of toxicity, efficacy, and surrogate efficacy.
Recently, many Bayesian methods have been developed for dose finding when simultaneously modeling both toxicity and efficacy outcomes in a blended phase I/II fashion. A further challenge arises when all the true efficacy data cannot be obtained quickly after the treatment so that surrogate markers are instead used (e.g., in cancer trials). We propose a framework to jointly model the probabilities of toxicity, efficacy, and surrogate efficacy given a particular dose. Our trivariate binary model is specified as a composition of two bivariate binary submodels. In particular, we extend the bivariate continual reassessment method (CRM), as well as utilize a particular Gumbel copula. The resulting trivariate algorithm utilizes all the available data at any given time point and can flexibly stop the trial early for either toxicity or efficacy. Our simulation studies demonstrate that our proposed method can successfully improve dosage targeting efficiency and guard against excess toxicity over a variety of true model settings and degrees of surrogacy.


Blacks in the Coronary Artery Surgery Study: risk factors and coronary artery disease.
In this paper we examine the relationship between risk factors and angiographically determined coronary artery disease for blacks and whites enrolled in the Coronary Artery Surgery Study (CASS). Analysis of data from the CASS registry indicated that blacks had a higher incidence of hypertension and current cigarette smoking than did whites in CASS and that chest pain was the major reason that both blacks and whites underwent coronary angiography for suspected or proven coronary disease. The CASS data also showed that, despite high levels of risk factors and chest pain, blacks had minimal or absent coronary disease. The results of this study raise several questions. First, to what extent are blacks in CASS representative of blacks in the general population and blacks undergoing coronary angiography? Additionally, are risk factors for coronary artery disease different for blacks than for whites? And finally, how does the physician effectively treat the black patient with high levels of risk factors and minimal coronary disease?


Pediatric intracranial gunshot wounds: the Memphis experience.
OBJECTIVE Penetrating brain injury in civilians is much less common than blunt brain injury but is more severe overall. Gunshot wounds (GSWs) cause high morbidity and mortality related to penetrating brain injury; however, there are few reports on the management and outcome of intracranial GSWs in children. The goals of this study were to identify clinical and radiological factors predictive for death in children and to externally validate a recently proposed pediatric prognostic scale. METHODS The authors conducted a retrospective review of penetrating, isolated GSWs sustained in children whose ages ranged from birth to 18 years and who were treated at 2 major metropolitan Level 1 trauma centers from 1996 through 2013. Several standard clinical, laboratory, and radiological factors were analyzed for their ability to predict death in these patients. The authors then applied the St. Louis Scale for Pediatric Gunshot Wounds to the Head, a scoring algorithm that was designed to provide rapid prognostic information for emergency management decisions. The scale's sensitivity, specificity, and positive and negative predictability were determined, with death as the primary outcome. RESULTS Seventy-one children (57 male, 14 female) had a mean age of 14 years (range 19 months to 18 years). Overall mortality among these children was 47.9%, with 81% of survivors attaining a favorable clinical outcome (Glasgow Outcome Scale score ≥ 4). A number of predictors of mortality were identified (all p < 0.05): 1) bilateral fixed pupils; 2) deep nuclear injury; 3) transventricular projectile trajectory; 4) bihemispheric injury; 5) injury to ≥ 3 lobes; 6) systolic blood pressure < 100 mm Hg; 7) anemia (hematocrit < 30%); 8) Glasgow Coma Scale score ≤ 5; and 9) a blood base deficit < -5 mEq/L. Patient age, when converted to a categorical variable (0-9 or 10-18 years), was not predictive. Based on data from the 71 patients in this study, the positive predictive value of the St. Louis scale in predicting death (score ≥ 5) was 78%. CONCLUSIONS This series of pediatric cranial GSWs underscores the importance of the initial clinical exam and CT studies along with adequate resuscitation to make the appropriate management decision(s). Based on our population, the St. Louis Scale seems to be more useful as a predictor of who will survive than who will succumb to their injury.


Towards fully automated structure-based NMR resonance assignment of ¹⁵N-labeled proteins from automatically picked peaks.
In NMR resonance assignment, an indispensable step in NMR protein studies, manually processed peaks from both N-labeled and C-labeled spectra are typically used as inputs. However, the use of homologous structures can allow one to use only N-labeled NMR data and avoid the added expense of using C-labeled data. We propose a novel integer programming framework for structure-based backbone resonance assignment using N-labeled data. The core consists of a pair of integer programming models: one for spin system forming and amino acid typing, and the other for backbone resonance assignment. The goal is to perform the assignment directly from spectra without any manual intervention via automatically picked peaks, which are much noisier than manually picked peaks, so methods must be error-tolerant. In the case of semi-automated/manually processed peak data, we compare our system with the Xiong-Pandurangan-Bailey-Kellogg's contact replacement (CR) method, which is the most error-tolerant method for structure-based resonance assignment. Our system, on average, reduces the error rate of the CR method by five folds on their data set. In addition, by using an iterative algorithm, our system has the added capability of using the NOESY data to correct assignment errors due to errors in predicting the amino acid and secondary structure type of each spin system. On a publicly available data set for human ubiquitin, where the typing accuracy is 83%, we achieve 91% accuracy, compared to the 59% accuracy obtained without correcting for such errors. In the case of automatically picked peaks, using assignment information from yeast ubiquitin, we achieve a fully automatic assignment with 97% accuracy. To our knowledge, this is the first system that can achieve fully automatic structure-based assignment directly from spectra. This has implications in NMR protein mutant studies, where the assignment step is repeated for each mutant.


A Novel Prognostic Predictor of Immune Microenvironment and Therapeutic Response in Kidney Renal Clear Cell Carcinoma based on Necroptosis-related Gene Signature.
Background: Necroptosis, a cell death of caspase-independence, plays a pivotal role in cancer biological regulation. Although necroptosis is closely associated with oncogenesis, cancer metastasis, and immunity, there remains a lack of studies determining the role of necroptosis-related genes (NRGs) in the highly immunogenic cancer type, kidney renal clear cell carcinoma (KIRC). Methods: The information of clinicopathology and transcriptome was extracted from TCGA database. Following the division into the train and test cohorts, a three-NRGs (TLR3, FASLG, ZBP1) risk model was identified in train cohort by LASSO regression. The overall survival (OS) comparison was conducted between different risk groups through Kaplan-Meier analysis, which was further validated in test cohort. The Cox proportional hazards regression model was introduced to assess its impact of clinicopathological factors and risk score on survival. ESTIMATE and CIBERSORT algorithms were introduced to evaluate immune microenvironment, while enrichment analysis was conducted to explore the biological significance. Correlation analysis was applied for the correlation assessment between checkpoint gene expression and risk score, between gene expression and therapeutic response. Gene expressions from TCGA were verified by GEO datasets and immunohistochemistry (IHC) analysis. Results: This NRGs-related signature predicted poorer OS in high-risk group, which was also verified in test cohort. Risk score could also independently predict survival outcome of KIRC. Significant changes were also found in immune microenvironment and checkpoint gene expressions between different risk groups, with immune functional enrichment in high-risk group. Interestingly, therapeutic response was correlated with the expressions of NRGs. The expressions of NRGs from TCGA were consistent with those from GEO datasets and IHC analysis. Conclusion: The NRGs-related signature functions as a novel prognostic predictor of immune microenvironment and therapeutic response in KIRC.


Percent Body Fat Measurement in the Medical Management of Children with Obesity.
Sustaining weight loss can be challenging, as physiological responses to weight loss, including metabolic and hormonal adaptations and decreased energy expenditure, promote weight regain. Paired with sustained dietary changes, physical activity can promote weight maintenance after successful weight loss, as physical activity can help maintain fat-free mass. We present several illustrative cases to highlight the potential use of body composition measurement using a bioelectrical impedance analysis (BIA) scale to augment obesity management counseling in a tertiary care pediatric weight-management clinic. BIA does require some interpretation, as it can be affected by hydration status and time of day, as well as patient age, sex, and body mass index. Nonetheless, BIA can be a helpful aid to obesity counseling. More research is needed to better understand how to use change in percent body fat over time as a motivational tool for management of children with obesity. [Pediatr Ann. 2018;47(12):e487-e493.].


An Aberrant Phosphorylation of Amyloid Precursor Protein Tyrosine Regulates Its Trafficking and the Binding to the Clathrin Endocytic Complex in Neural Stem Cells of Alzheimer's Disease Patients.
Alzheimer's disease (AD) is the most common cause of dementia and is likely caused by defective amyloid precursor protein (APP) trafficking and processing in neurons leading to amyloid plaques containing the amyloid-β (Aβ) APP peptide byproducts. Understanding how APP is targeted to selected destinations inside neurons and identifying the mechanisms responsible for the generation of Aβ are thus the keys for the advancement of new therapies. We previously developed a mouse model with a mutation at tyrosine (Tyr) 682 in the C-terminus of APP. This residue is needed for APP to bind to the coating protein Clathrin and to the Clathrin adaptor protein AP2 as well as for the correct APP trafficking and sorting in neurons. By extending these findings to humans, we found that APP binding to Clathrin is decreased in neural stem cells from AD sufferers. Increased APP Tyr phosphorylation alters APP trafficking in AD neurons and it is associated to Fyn Tyr kinase activation. We show that compounds affecting Tyr kinase activity and counteracting defects in AD neurons can control APP location and compartmentalization. APP Tyr phosphorylation is thus a potential therapeutic target for AD.


Assessment of molecularly imprinted sol-gel materials for selective room temperature phosphorescence recognition of nafcillin.
Sol-gel imprinted materials were prepared against nafcillin, a semisynthetic beta-lactamic antibiotic employed in the treatment of serious infections caused by penicillinase-producing staphylococci. Two approaches were addressed for preparation of the imprinted materials and the controls: as conventional monoliths, which were ground and sieved to a desired particle size for rebinding analysis, and as films on supporting glass slides. The specific binding sites that are created during the imprinting process are analyzed via selective room temperature phosphorescence (RTP) (sol-gel films) measurements as well as via competitive room temperature phosphorescence ligand assay. Results demonstrated the importance of the physical configuration of the imprinted material for minimizing non-specific binding. The close similarities between the structures of different beta-lactamic antibiotics made it possible to interpret the roles of the template structure on specific molecular recognition. In this article, we introduce the use of room temperature phosphorescence as selective transduction method for the template. The imprinted sol-gel films displayed enhanced specific binding characteristics respect to the monolithic sol-gel and can be envisaged for the use as recognition matrices for the screening and rapid selection of antibiotics from a combinatorial library or for the rapid control of nafcillin in biological samples (e.g. milk, serum, urine).


Early onset severe and late-onset mild Charcot-Marie-Tooth disease with mitofusin 2 (MFN2) mutations.
Mutations in the mitofusin 2 (MFN2) gene, which encodes a mitochondrial GTPase mitofusin protein, have recently been reported to cause both Charcot-Marie-Tooth 2A (CMT2A) and hereditary motor and sensory neuropathy VI (HMSN VI). It is well known that HMSN VI is an axonal CMT neuropathy with optic atrophy. However, the differences between CMT2A and HMSN VI with MFN2 mutations remained to be clarified. Therefore, we studied the phenotypic characteristics of CMT patients with MFN2 mutations. Mutations in MFN2 were screened in 62 unrelated axonal CMT neuropathy families. We calculated CMT neuropathy scores (CMTNSs) and functional disability scales (FDSs) to quantify disease severity. Twenty-one patients with the MFN2 mutations were studied by brain MRI. Ten pathogenic mutations were identified in 26 patients from 15 families (24.2%). Six of these mutations had not been reported, and de novo mutations were observed in five families (33.3%). The electrophysiological patterns of affected individuals with the MFN2 mutations were typical of axonal CMT; however, the clinical and electrophysiological characteristics were markedly different in early (<10 years) and late disease-onset (> or =10 years) groups. All patients with an early onset had severe CMTNS (> or =21) and FDS (6 or 7), whereas most patients with late onset had mild CMTNS (< or =10) and FDS (< or =3). We identified two HMSN VI families with the R364W mutation in the early onset group; however, two other families with the same mutation did not have optic atrophy. In addition, two early onset families with R94W mutations, previously reported for HMSN VI, did not have visual impairment. Interestingly, eight patients had periventricular and subcortical hyperintense lesions by brain MRI. In the late-onset group, three patients had sensorineural hearing loss and two had bilateral extensor plantar responses. We found that MFN2 mutations are the major cause of axonal CMT neuropathy, and that they are associated with variable CNS involvements. Phenotypes were significantly different in the early and late disease-onset groups. Our findings suggest that HMSN VI might be a variant of the early onset severe CMT2A phenotype.


Levels of uroporphyrinogen decarboxylase (URO-D) in erythrocytes of Italian porphyria cutanea tarda patients.
Porphyria cutanea tarda (PCT) is a human metabolic disorder due to the acquired or genetic impairment of uroporphyrinogen decarboxylase (URO-D) activity, the fifth enzyme of the heme biosynthetic pathway. A classification of inherited and non-inherited forms is based on the enzyme activity levels in red blood cells (RBC). Clinical manifestations of PCT are often precipitated by triggering factors such as alcohol, drug abuse, estrogens, virus infections, hepatotoxic chemicals and hepatic siderosis. We measured URO-D activity in RBC from a large sample of Italian PCT patients in order to define the enzyme activity distribution and to attempt a correlation among activity, risk factors and clinical outcome. Three classes of patients with low, normal and over-normal URO-D activity were defined according to control values. Low URO-D levels were present in 25.8% of patients, suggesting the familial form of PCT (type II). In this group, the outcome of PCT seems to be less influenced by risk factors. Patients with over-normal URO-D activity in RBC deserve further investigation.


Establishing health-promoting workplaces in Aboriginal community organisations: healthy eating policies.
Aboriginal community controlled health organisations (ACCHOs) and cooperatives function at the centre of community life for local Aboriginal people across Victoria. Local Aboriginal people govern them, work within them as managers and service providers, access health and community services from them and form the constituents who determine their directions. Victorian ACCHOs reflect the unique characteristics of the local Aboriginal community. Thus, potentially, Victorian ACCHOs are key strategic sites for health promotion activities that seek to establish and nurture healthy community, family and peer norms. The Victorian Aboriginal Community Controlled Health Organisation (VACCHO) partnered five metropolitan, regional and rural ACCHOs in a pilot project towards the establishment of healthy food policies and practices in their organisations. Project activities combined both 'top-down' policy-oriented and 'bottom-up' practice-oriented strategies. This paper, drawing upon both baseline and follow-up quantitative and qualitative data, describes initiatives leading to increases in healthy catering choices and related challenges for Aboriginal workplace health promotion practice.


Characterization of methicillin-resistant Staphylococcus aureus isolates from patients with persistent or recurrent bacteremia.
Methicillin-resistant Staphylococcus aureus (MRSA) bloodstream infections (BSI) are associated with considerable morbidity and mortality, especially with persistent (PB) or recurrent bacteremia (RB). To determine the frequency of PB and RB in patients with MRSA BSI, and to characterize the isolates from these patients. Surveillance for MRSA BSI was performed for one year in 13 Canadian hospitals. PB was defined as a positive blood culture that persisted for ≥7 days; RB was defined as the recurrence of a positive blood culture ≥14 days following a negative culture. Isolates were typed using pulsed-field gel electrophoresis (PFGE). Vancomycin susceptibility was determined using Etest. A total of 183 patients with MRSA BSI were identified; 14 (7.7%) had PB and five (2.7%) had RB. Ten (5.5%) patients were known to have infective endocarditis, and five of these patients had PB or RB. Initial and subsequent MRSA isolates from patients with PB and RB had the same PFGE type. There were no significant differences in the distribution of PFGE types in patients with PB or RB (37% CMRSA-2/USA100; 37% CMRSA-10/USA300) compared with that in other patients (56% CMRSA-2/USA100; 32% CMRSA-10/USA300). All isolates were susceptible to vancomycin, but patients with PB or RB were more likely to have initial isolates with vancomycin minimum inhibitory concentration = 2.0 μg/mL (26% versus 10%; P=0.06). Persistent or recurrent MRSA bacteremia occurred in 10.4% of patients with MRSA BSIs. Initial isolates from patients with persistent or recurrent MRSA BSIs were more likely to exhibit reduced susceptibility to vancomcyin, but were not associated with any genotype.


A Novel Interleukin 17 Receptor A Mutation in a Child with Chronic Mucocutaneous Candidiasis and Staphylococcal Skin Infections.
Chronic mucocutaneous candidiasis leads to persistent or recurrent fungal infections of the nail, skin, oral, and genital mucosa. Impaired interleukin 17-mediated immunity is a cause of chronic mucocutaneous candidiasis. We aimed to show the pathogenicity of a novel interleukin 17 receptor A mutation through functional studies. After next-generation sequencing analysis showed the interleukin 17 receptor A variant, we confirmed the variant by Sanger sequencing and functional validation of the variant by flow cytometry. We present the case of a 6-year-old male patient who presented with recurrent oral and genital Candida infections and eczema. He had staphylococcal skin lesions, fungal susceptibility, and eczema. The patient carried a novel homozygous nonsense [(c.787C> T) (p.Arg263Ter)] mutation in the interleukin 17 receptor A gene. Sanger sequencing confirmed the variant and revealed the segregation of the variant in the family. We used flow cytometry to detect interleukin 17 receptor A protein expression in peripheral blood mononuclear cells from patients and measured Th17 cell percentage. We observed low interleukin 17 receptor A protein expression in patient peripheral blood mononuclear cells, decreased CD4+ interleukin 17+ cell percentage, and decreased interleukin 17F expression in CD4+ cells compared to healthy controls. Innate immune defects may lead to chronic recurrent fungal and bacterial infections of the skin, mucosa, and nails. Generally, genetic and functional analysis is needed in addition to basic immunological tests.


Experimental intramammary inoculation with Mycoplasma bovis in vaccinated and unvaccinated cows: effect on the mycoplasmal infection and cellular inflammatory response.
The effect of vaccination on mycoplasmal infection and the cellular inflammatory response was evaluated in 4 vaccinated and 4 control cows experimentally challenged in 2 of 4 quarters with live Mycoplasma bovis. In unchallenged quarters during the first three weeks after experimental challenge exposure, 6 of 8 quarters on control cows, and 7 of 8 quarters on vaccinated cows became infected with low numbers (10(2)-10(4) cfu/ml) of M bovis. During the same period all challenge-infused quarters on both control and vaccinated animals became infected with high numbers (10(9) cfu/ml) of M bovis. Thereafter, all quarters on vaccinated cows became culture-negative for M bovis, while 2 of 8 unchallenged quarters, and 4 of 8 challenged quarters on 3 of 4 control cows remained infected. A cellular inflammatory response as measured by the California Mastitis Test accompanied the experimental infection in proportion to the infection level except in challenged quarters on vaccinated cows after the first three weeks post challenge in which the cellular inflammatory response remained high despite the advent of negative M bovis culture results. This study indicates that the course of experimental M bovis mastitis can be affected by vaccination, and that vaccination results in an adverse cellular inflammatory response in challenged quarters.


Diagnostic validity of the hyperactive child (attention deficit disorder with hyperactivity) syndrome.
The evolution of the hyperactive child syndrome as a diagnostic category is traced through various pseudo-etiological formulations (minimal brain dysfunction) to the current descriptive DSM III category as ADDH. The review suggests that the use of inconsistent and poorly specified selection criteria in many studies has led to confusion in terminology, and controversy over the diagnostic validity of the syndrome. Probably because of diagnostic difficulties, biological and genetic studies have not so far definitively validated a syndrome distinct from conduct and learning disorders. Among the most important questions to be answered is whether response to psychostimulants depends on a unique and qualitatively different action in ADDH children. The syndrome is undoubtedly not homogeneous and recent attempts to isolate subgroups on the basis of independent hyperactivity-inattention and aggressive dimensions have shown promise in predicting natural history and therapeutic response. Thus improved methods of subclassification, coupled with improved measures of activity level and attention span are required.


Prescription opiate analgesics, heroin, HIV and HCV among persons who inject drugs in New York City, 2016-2018.
Assess relationships among non-medical use of prescription opioid analgesics (POAs), heroin use, and HIV and hepatitis C (HCV) infection among persons who inject drugs (PWID) in New York City, 2016-2018. PWID (N = 134) were recruited from Mount Sinai Beth Israel drug treatment programs. HIV seropositive persons were oversampled. A questionnaire was administered, and serum samples were collected for HIV and HCV testing. Analyses were stratified by HIV serostatus and compared those who had used POAs to those who had not used POAs. Among the participants, 97% reported injecting heroin, 44% reported injecting cocaine, and 47% reported smoking crack cocaine in the 6 months prior to the interview. There were 66% who reported oral non-medical use of POAs, with 42% using oral POAs in the previous 6 months. There was a clear historical pattern in median year of first injection for different groups: HIV seropositive persons (1985), HIV seronegative persons who never used POAs (1999), and HIV seronegative persons who used POAs (2009). By the time of interview (2016-2018), however, almost all participants (97%) reported injecting heroin. All PWID who reported using POAs also reported injecting heroin. Non-medical POA use among PWID was very common and should not be considered a separate drug use epidemic, but as an additional component of the continuing heroin/poly-drug use epidemic, itself a part of the syndemic of opioid use, stimulant use, overdose, HCV and HIV occurring in New York City.


Effect of the usage of Er,Cr:YSGG laser with and without different remineralization agents on the enamel erosion of primary teeth.
The aim of the present study was to evaluate the effects of different remineralization agents associated with erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) (0.5 W power, 20 Hz frequency, 60% water, 40% air, 25 mJ pulse energy, 8.84 J/cm2 fluence, 60 μs pulse duration, 600 μm tip diameter, and an approximate 1-1.5 mm distance to the target) laser irradiation on erosion induced by the consumption of carbonated drinks in human primary enamel. There were 8 groups and 10 primary teeth in each g0roup. The distribution was as follows: group 1, casein phosphopeptide-amorphous calcium phosphate with fluoride (CPP-ACPF); group 2, Er,Cr:YSGG laser+CPP-ACPF; group 3, fluor varnish; group 4, Er,Cr:YSGG Laser+fluoride varnish; group 5, ROCS® medical mineral gel; group 6, Er,Cr:YSGG laser + ROCS® medical mineral gel; group 7, Er,Cr:YSGG laser; and group 8, artificial saliva. The samples in the groups were submerged in artificial saliva and acid twice a day for 6 s at 6-h intervals and were then exposed to an erosion cycle 15 times. In the groups in which the Er,Cr:YSGG laser was applied in combination with the remineralization agents, the laser application was made first, and then the remineralization agents were applied for 4 min in each group. The Friedman and Wilcoxon signed-rank tests and the Bonferroni correction were used in statistical analyses, and the significance level was taken as p < 0.05. According to the results, all agents had a statistically significant difference (groups 1, 2, 3, 4, and 6: p = 0.005, p < 0.017; groups 5 and 7: p = 0.007, p < 0.017) between BL-RM periods. However, all agents had a statistically significant remineralization effect on primary teeth enamel (groups 1, 2, 3, 6, and 7: p = 0.005, p < 0.017; group 4: p = 0.011, p < 0.017) except that group 5 (p = 0.074, p < 0.017) between DM-RM periods. The coadministration of an agent with the laser did not make any difference at a statistical level (p = 0.804, p > 0.05). The results were supported by scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis. As a result of this study, CPP-ACPF had a notable impact in terms of the remineralization effect on eroded enamel, and the Er,Cr:YSGG laser alone may be an alternative method, which may be related to the modified hydroxyapatite structure, 38.5% H0.56Ca4.56O13P3Y0.44, that was determined in XRD analysis.


Autonomous targeting of infectious superspreaders using engineered transmissible therapies.
Infectious disease treatments, both pharmaceutical and vaccine, face three universal challenges: the difficulty of targeting treatments to high-risk 'superspreader' populations who drive the great majority of disease spread, behavioral barriers in the host population (such as poor compliance and risk disinhibition), and the evolution of pathogen resistance. Here, we describe a proposed intervention that would overcome these challenges by capitalizing upon Therapeutic Interfering Particles (TIPs) that are engineered to replicate conditionally in the presence of the pathogen and spread between individuals--analogous to 'transmissible immunization' that occurs with live-attenuated vaccines (but without the potential for reversion to virulence). Building on analyses of HIV field data from sub-Saharan Africa, we construct a multi-scale model, beginning at the single-cell level, to predict the effect of TIPs on individual patient viral loads and ultimately population-level disease prevalence. Our results show that a TIP, engineered with properties based on a recent HIV gene-therapy trial, could stably lower HIV/AIDS prevalence by ∼30-fold within 50 years and could complement current therapies. In contrast, optimistic antiretroviral therapy or vaccination campaigns alone could only lower HIV/AIDS prevalence by <2-fold over 50 years. The TIP's efficacy arises from its exploitation of the same risk factors as the pathogen, allowing it to autonomously penetrate superspreader populations, maintain efficacy despite behavioral disinhibition, and limit viral resistance. While demonstrated here for HIV, the TIP concept could apply broadly to many viral infectious diseases and would represent a new paradigm for disease control, away from pathogen eradication but toward robust disease suppression.


Dizziness and Imbalance in the Elderly: Age-related Decline in the Vestibular System.
Dizziness and imbalance are amongst the most common complaints in older people, and are a growing public health concern since they put older people at a significantly higher risk of falling. Although the causes of dizziness in older people are multifactorial, peripheral vestibular dysfunction is one of the most frequent causes. Benign paroxysmal positional vertigo is the most frequent form of vestibular dysfunction in the elderly, followed by Meniere's disease. Every factor associated with the maintenance of postural stability deteriorates during aging. Age-related deterioration of peripheral vestibular function has been demonstrated through quantitative measurements of the vestibulo-ocular reflex with rotational testing and of the vestibulo-collic reflex with testing of vestibular evoked myogenic potentials. Age-related decline of vestibular function has been shown to correlate with the age-related decrease in the number of vestibular hair cells and neurons. The mechanism of age-related cellular loss in the vestibular endorgan is unclear, but it is thought that genetic predisposition and cumulative effect of oxidative stress may both play an important role. Since the causes of dizziness in older people are multi-factorial, management of this disease should be customized according to the etiologies of each individual. Vestibular rehabilitation is found to be effective in treating both unilateral and bilateral vestibular dysfunction. Various prosthetic devices have also been developed to improve postural balance in older people. Although there have been no medical treatments improving age-related vestibular dysfunction, new medical treatments such as mitochondrial antioxidants or caloric restriction, which have been effective in preventing age-related hearing loss, should be ienvestigated in the future.


Human papillomavirus infection: etiopathogenesis, molecular biology and clinical manifestations.
Human papillomavirus (HPV) is a DNA virus that presents tropism for epithelial cells, causing infections of the skin and mucous membranes. Replication of HPV occurs in the nuclei of squamous cells and its life cycle is directly related to the differentiation program of the host cell. To date, nearly 100 different types of HPV have been characterized and there is a large number of other types that have not been sequenced yet. Besides being responsible for benign lesions of the skin and mucous membranes, HPV is also involved in the development of various mucocutaneous tumors: Bowen's disease, non-melanoma skin cancers and genital carcinomas. This review discusses the characteristics of HPV, malignant and benign mucous and skin manifestations caused by HPV, besides the main methods of detection and typing of the virus.


Upregulation of miR181a/miR212 Improves Myogenic Commitment in Murine Fusion-Negative Rhabdomyosarcoma.
Fusion-negative rhabdomyosarcoma (FN-RMS) is the most common soft tissue sarcoma of childhood arising from undifferentiated skeletal muscle cells from uncertain origin. Currently used therapies are poorly tumor-specific and fail to tackle the molecular machinery underlying the tumorigenicity and uncontrolled proliferation of FN-RMS. We and other groups recently found that microRNAs (miRNA) network contributes to myogenic epigenetic memory and can influence pluripotent stem cell commitments. Here, we used the previously identified promyogenic miRNAs and tailored it to the murine FN-RMS. Subsequently, we addressed the effects of miRNAs in vivo by performing syngeneic transplant of pre-treated FN-RMS cell line in C57Bl/6 mice. miRNA pre-treatment affects murine FN-RMS cell proliferation in vivo as showed by bioluminescence imaging analysis, resulting in better muscle performances as highlighted by treadmill exhaustion tests. In conclusion, in our study we identified a novel miRNA combination tackling the anti-myogenic features of FN-RMS by reducing proliferation and described novel antitumorigenic therapeutic targets that can be further explored for future pre-clinical applications.


Novel classification based on immunohistochemistry combined with hierarchical clustering analysis in non-functioning neuroendocrine tumor patients.
Somatostatin analogues ameliorated many symptoms caused by neuroendocrine tumors (NET), but their antitumor activities are limited especially in non-functioning cases. An overactivation of signaling pathways under receptor tyrosine-kinase (RTK) has been recently demonstrated in some NET patients, but its details have remained largely unknown. Therefore, in this study, we immunolocalized therapeutic factors and evaluated the data to study the clinical significance of the molecules in non-functioning Japanese gastrointestinal NET. Fifty-two NET cases were available for examination in this study and expression of somatostatin receptor (sstr) 1, 2A, 2B, 3 and 5, activated form of mammalian target of rapamycin (mTOR), eukaryotic initiation factor 4-binding protein 1 (4EBP1), ribosomal protein s6 (S6), extracellular signal-regulated kinase (ERK) and insulin-like growth factor 1 receptor (IGF-1R) was evaluated using immunohistochemistry. We then studied the correlation among the immunohistochemical results of the individual cases using hierarchical clustering analysis. Results of clustering analysis demonstrated that NET cases were basically classified into Cluster I and II. Cluster I was associated with higher expression of sstr1, 2B and 3 and Cluster II was characterized by an activation of the PI3K/Akt pathway and IGF-1R and higher proliferative status. Cluster II was further classified into Cluster IIa and IIb. Cluster IIa was associated with higher expression of sstr1 and 5 and higher proliferative status and Cluster IIb was characterized by ERK activation. Hierarchical clustering analysis of immunoreactivity of the therapeutic factors can classify NET cases into three distinctive groups and the medical treatment may be determined according to this novel classification method for non-functioning NET patients.


Think twice before re-manipulating distal metaphyseal forearm fractures in children.
Treatment of displaced paediatric distal forearm fractures is not always successful. Re-occurrence of angular deformity is a frequent complication. No consensus exists when to perform secondary manipulations. The purpose of this study was to analyse the long-term outcome of re-angulated paediatric forearm fractures to determine if re-manipulations can be avoided. Children who underwent closed reduction for distal forearm fractures and presented with re-angulation at follow-up were included in this retrospective cohort study. We compared those that were re-manipulated to those managed conservatively. Re-angulation was defined as ≥15° of angulation on either the AP or lateral view. Children were reviewed after 1-8 years post injury. Outcome measures were residual angulation on radiographs, active range of motion, grip strength, Visual Analogue Scales (satisfaction, cosmetics and pain) and the ABILHANDS-kids questionnaire. Sixty-six children (mean age of 9.6 years) were included. Twenty-four fractures were re-manipulated and 42 fractures had been left to heal in angulated position. At time of re-angulation, children <12 years in the conservative group had similar angulations to those re-manipulated. Children ≥12 years in the re-manipulation group had significantly greater angulations than children in the conservative group. At final follow-up, after a mean of 4.0 years, near anatomical alignment was seen on radiographs in all patients. Functional outcome was predominantly excellent. There was no significant difference in functional, subjective or radiological outcomes between treatment groups. Re-manipulation of distal forearm fractures in children <12 years did not improve outcomes, deeming re-manipulations unnecessary. Children ≥12 years in the conservative group achieved satisfactory outcomes despite re-angulations exceeding current guidelines. Based on observed remodelling, we now accept up to 30° angulation in children <9 years; 25° angulation in children aged 9-<12; 20° angulation in children ≥12 years, when re-angulation occurs. We conclude that clinicians should be more reluctant to perform re-manipulations.


Safety and efficacy of CDX-014, an antibody-drug conjugate directed against T cell immunoglobulin mucin-1 in advanced renal cell carcinoma.
CDX-014 is an antibody-drug conjugate directed against TIM-1, a surface marker highly expressed in renal cell carcinoma (RCC) and ovarian carcinoma. This phase I, first-in-human trial was conducted to evaluate the safety and preliminary activity of CDX-014 in patients with advanced refractory RCC, following a dose-escalation and dose expansion design. CDX-014 was administered intravenously at doses ranging from 0.15 to 2.0 mg/kg every 2 or 3 weeks until progression or unacceptable toxicity. Sixteen patients received at least one dose of CDX-014. The maximum tolerated dose was not identified. Most frequent adverse grade 1 or 2 adverse events included nausea (38%), fatigue, alopecia, elevation of AST and decreased appetite (25% each). Adverse events of grade 3 or more included hyperglycemia (19%), urosepsis (6%), and one multi-organ failure (6%) responsible for one treatment-related death. Two patients discontinued therapy for adverse events including fatigue grade 2 and urosepsis grade 4. CDX-014 showed antitumor activity with one prolonged partial response and a clinical benefit rate (objective response or stable disease >6 months) of 31%. The two patients that exhibited the most marked tumor shrinkage had high TIM-1 expression on tumor tissue. Overall, CDX-014 exhibited a manageable toxicity profile and early signs of activity, supporting further evaluation of antibody-drug conjugates in patients with advanced RCC and potentially other TIM-1 expressing cancers. Trial registration https://clinicaltrials.gov/ct2/show/NCT02837991 NCT02837991; July 20, 2016.


Estrogen-progesterone balance in the context of blastocyst implantation failure induced by predator stress.
Diverse stressors can disrupt blastocyst implantation in inseminated female mammals. Stress-induced implantation failure can be mimicked by minute doses of exogenous estradiol, and some evidence indicates that it may be mitigated by exogenous progesterone. In Experiment 1, we showed that acute exposure to a rat across a wire-mesh grid caused elevation of corticosterone and progesterone. In Experiment 2, we showed that exposure of inseminated mice to rats across a grid during gestation days 1-5 was associated with avoidance of proximity to the grid and a significantly reduced number of implantation sites on gestation day 6. Rat-exposure also resulted in elevated progesterone levels in females that maintained their pregnancies, and elevated estradiol levels in females that lost their pregnancies. In Experiment 3, we investigated whether exogenous progesterone, estradiol, or a combination of both could influence implantation failure induced by rat-exposure stress. Treatment with 100 ng estradiol per day on gestation days 1-5 induced a complete absence of implantation sites on gestation day 6, regardless of the presence or absence of the stressor. Administration of 500 μg progesterone per day was insufficient to prevent the stress-induced pregnancy loss. However, 500 μg progesterone plus 10 ng estradiol per day did prevent implantation failure in rat-exposed females. These findings are consistent with the hypothesis that estradiol elevations contribute to stress-induced pregnancy loss, but show paradoxically that low doses of estradiol can act together with progesterone to mitigate stress-induced pregnancy loss.


MCNPX CALCULATIONS OF SPECIFIC ABSORBED FRACTIONS IN SOME ORGANS OF THE HUMAN BODY DUE TO APPLICATION OF 133Xe, 99mTc and 81mKr RADIONUCLIDES.
Monte Carlo simulations were performed to evaluate treatment doses with wide spread used radionuclides 133Xe, 99mTc and 81mKr. These different radionuclides are used in perfusion or ventilation examinations in nuclear medicine and as indicators for cardiovascular and pulmonary diseases. The objective of this work was to estimate the specific absorbed fractions in surrounding organs and tissues, when these radionuclides are incorporated in the lungs. For this purpose a voxel thorax model has been developed and compared with the ORNL phantom. All calculations and simulations were performed by means of the MCNP5/X code.


[Retrogastral located gastrointestinal stromal tumor (GIST) as a sonographically detected rare incidental finding].
Gastrointestinal stromal tumors (GIST) represent compared to carcinomas a rare group of neoplasias of the gastro-intestinal tract of unclear dignity. We report the example of a patient suffering from a big retrogastral located gastrointestinal stromal tumor which had been detected as an incidental finding without previous complaints. Because origin and dignity of the process could not definitely be diagnosed, total resection (R0-resection) without systemic lymphadenectomy of the process measuring 11.5 cm x 11 cm x 7 cm was performed. Mitotic activity and tumor-size are regarded as predictive factors of potenzial malignancy of GISTs. In general tumors with low mitotic activity of up to 5 mitoses per 50 high power fields (HPFs) and a diameter smaller than 5 cm are regarded as benign. In the presented case, up to 4 mitoses per 50 HPFs could be detected and thus, in connection with tumor-size, an uncertain biological behaviour of the process has to be expected. Since no generally accepted consensus on the treatment of the GISTs exists, also patients originally suffering from tumors regarded as borderline-malignant should undergo a close-meshed follow-up in regular intervals.


Octave-spanning single-cycle middle-infrared generation through optical parametric amplification in LiGaS2.
We report the generation of extremely broadband and inherently phase-locked mid-infrared pulses covering the 5 to 11 µm region. The concept is based on two stages of optical parametric amplification starting from a 270-fs Yb:KGW laser source. A continuum seeded, second harmonic pumped pre-amplifier in β-BaB2O4 (BBO) produces tailored broadband near-infrared pulses that are subsequently mixed with the fundamental pump pulses in LiGaS2 (LGS) for mid-infrared generation and amplification. The pulse bandwidth and chirp is managed entirely by selected optical filters and bulk material. We find an overall quantum efficiency of 1% and a mid-infrared spectrum smoothly covering 5-11 µm with a pulse energy of 220 nJ at 50 kHz repetition rate. Electro-optic sampling with 12-fs long white-light pulses directly from self-compression in a YAG crystal reveals near-single-cycle mid-infrared pulses (32 fs) with passively stable carrier-envelope phase. Such pulses will be ideal for producing attosecond electron pulses or for advancing molecular fingerprint spectroscopy.


Advances in bio-optical imaging for the diagnosis of early oral cancer.
Oral cancer is among the most common malignancies worldwide, therefore early detection and treatment is imperative. The 5-year survival rate has remained at a dismal 50% for the past several decades. The main reason for the poor survival rate is the fact that most of the oral cancers, despite the general accessibility of the oral cavity, are not diagnosed until the advanced stage. Early detection of the oral tumors and its precursor lesions may be the most effective means to improve clinical outcome and cure most patients. One of the emerging technologies is the use of non-invasive in vivo tissue imaging to capture the molecular changes at high-resolution to improve the detection capability of early stage disease. This review will discuss the use of optical probes and highlight the role of optical imaging such as autofluorescence, fluorescence diagnosis (FD), laser confocal endomicroscopy (LCE), surface enhanced Raman spectroscopy (SERS), optical coherence tomography (OCT) and confocal reflectance microscopy (CRM) in early oral cancer detection. FD is a promising method to differentiate cancerous lesions from benign, thus helping in the determination of adequate resolution of surgical resection margin. LCE offers in vivo cellular imaging of tissue structures from surface to subsurface layers and has demonstrated the potential to be used as a minimally invasive optical biopsy technique for early diagnosis of oral cancer lesions. SERS was able to differentiate between normal and oral cancer patients based on the spectra acquired from saliva of patients. OCT has been used to visualize the detailed histological features of the oral lesions with an imaging depth down to 2-3 mm. CRM is an optical tool to noninvasively image tissue with near histological resolution. These comprehensive diagnostic modalities can also be used to define surgical margin and to provide a direct assessment of the therapeutic effectiveness.


Handle with care: A systematic review on frailty in cardiac care and its usefulness in heart transplantation.
There is growing consensus that frailty, a state of vulnerability and a decline in functioning across multiple physiological body systems, is a valuable criterion to guide clinicians' risk prediction for poor outcomes in adult transplant candidates. In its 2016 listing criteria for heart transplantation the International Society for Heart Lung Transplantation recommends frailty assessment. We aimed to summarize the usefulness of frailty assessment in heart transplant candidates or recipients reported throughout the literature. We performed a systematic literature search in PubMed to identify papers reporting on frailty in transplantation, chronic heart failure, and ventricualr assist device implantation published over the last 10 years in English. Additionally, a hand search was conducted, including manually searching the reference lists and a citation search of relevant papers. Eleven primary research articles were included in this systematic review. Frailty is a risk factor for morbidity, hospitalization, and mortality in patients with advanced heart failure and individuals being considered for ventricualr assist device implantation. Of the patients being considered for transplantation, 33% are frail. The Frailty Phenotype by Fried is a particularly useful tool to quickly identify higher risk patients for adverse outcomes. A lack of standardization and limited evidence on frailty in transplantation limit its use as a definitive listing criterion. Future research efforts should focus on systematic integration of frailty measures in transplant practice.


Establishment of the DU.528 human lymphohemopoietic stem cell line.
We have established the DU.528 cell line from the pretreatment leukemia cells of a patient who underwent a T lymphoblastic-to-promyelocytic phenotype conversion during treatment with the adenosine deaminase inhibitor, deoxycoformycin. The cell line and clones obtained from it by limiting dilution have the same karyotype previously found in the patient's pretreatment T lymphoblasts and post-deoxycoformycin treatment promyelocytes. DU.528 cells in continuous culture for greater than 2 yr display a predominant undifferentiated T lymphoblastoid phenotype. These cells spontaneously generate progeny of at least three lineages, T lymphoid, granulocytic/monocytic, and erythroid. The surface marker most consistently expressed by DU.528 cells in the undifferentiated state is the 3A1 antigen, which has been found on prothymocytes in the embryonic thymus. Some undifferentiated DU.528 cells also expressed the IL-2 receptor, but no other T cell differentiation antigens. Exposure of DU.528 cells to a variety of agents induced myeloid maturation; adenosine and deoxyadenosine, in the presence of deoxycoformycin, induced expression of myeloid differentiation antigens. Our results suggest that DU.528 is a lymphohematopoietic stem cell line and support the hypothesis that differentiation of pluripotent stem cells may be altered by genetic deficiency of adenosine deaminase. DU.528 cells may provide a useful model for examining factors that regulate stem cell proliferation and differentiation.


Comparison of the Haas and the Oxford classifications for prediction of renal outcome in patients with IgA nephropathy.
Pathologic features can provide valuable information for determining prognosis in IgA nephropathy (IgAN). However, it is uncertain whether the Oxford classification, a new classification of IgAN, can predict renal outcome better than previous ones. We conducted a retrospective cohort study in 500 patients with biopsy-proven IgAN between January 2002 and December 2010 to compare the ability of the Haas and the Oxford classifications to predict renal outcome. Primary outcome was a doubling of the baseline serum creatinine concentration (D-SCr). During a mean follow-up of 68 months, 52 (10.4%) and 35 (7.0%) developed D-SCr and end-stage renal disease, respectively. There were graded increases in the development of D-SCr in the higher Haas classes. In addition, the primary endpoint of D-SCr occurred more in patients with the Oxford M and T lesions than those without such lesions. In multivariate Cox regression analyses, the Haas class V (HR, 12.19; P=.002) and the Oxford T1 (hazard ratio [HR], 6.68; P<.001) and T2 (HR, 12.16; P<.001) lesions were independently associated with an increased risk of reaching D-SCr. Harrell's C index of each multivariate model with the Haas and the Oxford classification was 0.867 (P=.015) and 0.881 (P=.004), respectively. This was significantly higher than that of model with clinical factors only (C=0.819). However, there was no difference in C-statistics between the 2 models with the Haas and the Oxford classifications (P=.348). This study suggests that the Haas and the Oxford classifications are comparable in predicting progression of IgAN.


Function in athymic nude mice of parathyroid heterografts from patients with primary hyperparathyroidism and secondary hyperparathyroidism.
Heterotransplantation of adenomatous parathyroid glandular tissue from humans with primary hyperparathyroidism into athymic nude mice creates a unique animal model of this disease. The mice manifest high concentrations of both midregion/C-terminal human parathyroid hormone and biologically active intact human parathyroid hormone relative to either mice with no implants or mice that received normal human parathyroid tissue. Secretion of these substances is maintained in most mice for at least 9 to 13 months after implantation. In addition, animals that have experienced implantation exhibit other characteristics associated with human primary hyperparathyroidism including relative hypercalcemia and increased renal 25-hydroxyvitamin D-1 alpha-hydroxylase activity. We also measured these parameters in a group of nude mice that received transplantation of a similar mass of hyperplastic parathyroid tissue that was obtained from patients with uremic secondary hyperparathyroidism. Although we hypothesized that the level of human parathyroid hormone secretion from these implants would fall over time in response to the normal host environment, hormone levels remained as high as those in recipients of adenomatous heterografts, even after 9 to 13 months. Moreover, similar biologic effect of the excess parathyroid hormone (i.e., relative hypercalcemia, hyperphosphatasemia, and increased 1,25-dihydroxyvitamin D biosynthesis) were detected. These animal models should prove extremely useful in supplementing our understanding of hyperparathyroid disorder in man.


Quantitation of human cytochrome P450 2D6 protein with immunoblot and mass spectrometry analysis.
Accurate quantification of cytochrome P450 (P450) protein contents is essential for reliable assessment of drug safety, including the prediction of in vivo clearance from in vitro metabolism data, which may be hampered by the use of uncharacterized standards and existence of unknown allelic isozymes. Therefore, this study aimed to delineate the variability in absolute quantification of polymorphic CYP2D6 drug-metabolizing enzyme and compare immunoblot and nano liquid chromatography coupled to mass spectrometry (nano-LC/MS) methods in identification and relative quantification of CYP2D6.1 and CYP2D6.2 allelic isozymes. Holoprotein content of in-house purified CYP2D6 isozymes was determined according to carbon monoxide difference spectrum, and total protein was quantified with bicinchoninic acid protein assay. Holoprotein/total CYP2D6 protein ratio was markedly higher for purified CYP2D6.1 (71.0%) than that calculated for CYP2D6.1 Supersomes (35.5%), resulting in distinct linear calibration range (0.05-0.50 versus 0.025-0.25 pmol) that was determined by densitometric analysis of immunoblot bands. Likewise, purified CYP2D6.2 and CYP2D6.10 and the CYP2D6.10 Supersomes all showed different holoprotein/total CYP2D6 protein ratios and distinct immunoblot linear calibration ranges. In contrast to immunoblot, nano-LC/MS readily distinguished CYP2D6.2 (R296C and S486T) from CYP2D6.1 by isoform-specific proteolytic peptides that contain the altered amino acid residues. In addition, relative quantitation of the two allelic isozymes was successfully achieved with label-free protein quantification, consistent with the nominated ratio. Because immunoblot and nano-LC/MS analyses measure total P450 protein (holoprotein and apoprotein) in a sample, complete understanding of holoprotein and apoprotein contents in P450 standards is desired toward reliable quantification. Our data also suggest that nano-LC/MS not only facilitates P450 quantitation but also provides genotypic information.


Cloning and genetic organization of the pca gene cluster from Acinetobacter calcoaceticus.
The beta-ketoadipate pathway of Acinetobacter calcoaceticus comprises two parallel metabolic branches. One branch, mediated by six enzymes encoded by the cat genes, converts catechol to succinate and acetyl coenzyme A (acetyl-CoA); the other branch, catalyzed by products of the pca genes, converts protocatechuate to succinate and acetyl-CoA by six metabolic reactions analogous or identical to those of the catechol sequence. We used the expression plasmid pUC18 to construct expression libraries of DNA from an A. calcoaceticus mutant strain from which the cat genes had been deleted. Immunological screening with antiserum to the pcaE gene product, beta-ketoadipate:succinyl-CoA transferase I, resulted in the isolation of a cloned 11-kilobase-pair (kbp) fragment which inducibly expressed all six pca genes under control of the lac promoter on pUC18. The induced Escherichia coli cells formed the six pca gene products at levels 10- to 30-fold higher than found in fully induced A. calcoaceticus cultures, although protocatechuate 3,4-dioxygenase (the iron-containing product of the pcaA gene) from the recombinant strain possessed a relatively low turnover number. An E. coli culture expressing the cloned pca genes quantitatively converted protocatechuate to beta-ketoadipate; failure of the organism to metabolize the latter compound can be most readily ascribed to relatively low pool levels of succinyl-CoA, a required substrate for beta-ketoadipate:succinyl-CoA transferase, in E. coli. The gene order and direction of transcription were determined to be pcACBDFE by identification of enzymes expressed in subclones, by using natural transformation to identify subclones carrying DNA corresponding to dysfunctional alleles in mutant A. calcoaceticus strains, and by restriction mapping of both the 11-kbp fragment and derivatives of the 11-kbp fragment containing Tn5 in the pcaA, pcaB, pcaD, and pcaE genes. The fragment containing the pca gene hybridized strongly and specifically to a previously cloned fragment containing A. calcoaceticus cat genes.


Effect of atrial natriuretic peptide on vasa recta blood flow in the rat.
To determine whether synthetic atrial natriuretic peptide (ANP) increases renal medullary blood flow and if so whether the increase mediates the diuresis and natriuresis induced by ANP, inner medullary vasa recta blood flow in the exposed left renal papilla of anesthetized Munich Wistar rats weighing between 102 and 161 g was measured by fluorescence videomicroscopy. The rats were maintained in a euvolemic state by the infusion of albumin. Synthetic ANP (Auriculin B) was administered intravenously as 2.5 micrograms/kg body wt prime and as a continuous infusion of 0.2 microgram X min-1 X kg body wt-1 to the experimental group (n = 7). Within 2 min after ANP was given, urine flow and sodium excretion increased (29.4 +/- 3.8 to 50.4 +/- 5.8 microliter X min-1 X kidney wt-1, P less than 0.01, and 3.39 +/- 0.57 to 6.05 +/- 0.95 mueq X min-1 X g kidney wt-1, P less than 0.01, respectively), but vasa recta blood flow in descending (DVR) or ascending (AVR) vasa recta did not change significantly (9.5 +/- 2.3 to 10.0 +/- 2.8 nl/min in DVR and 5.3 +/- 1.0 to 6.1 +/- 1.2 nl/min in AVR). Forty-five minutes after ANP was begun, urine flow and sodium excretion increased further (77.1 +/- 11.1 microliter X min-1 X g kidney wt-1 and 12.0 +/- 2.15 mueq X min-1 X g kidney wt-1, respectively), and by this time vasa recta blood flow had increased significantly to 14.0 +/- 2.6 in DVR, P less than 0.01, and 9.8 +/- 1.2 in AVR, P less than 0.01.(ABSTRACT TRUNCATED AT 250 WORDS)


A history of medical professionalisation in NSW: 1788-1950.
This paper presents a brief history of medical professionalisation in the colony and State of New South Wales to the middle of the twentieth century. It is suggested initially that although 'western' medicine is a powerful international organisation with common structures and a shared scientific core, any worthwhile study of the medical profession in a particular setting must be founded on an analysis of the intersection of local conditions and international structures. Thus various overseas and interstate accounts of the social history of medicine are reviewed in terms of their relevance in helping to explain the process of medical professionalisation as it occurred in New South Wales.


Solidity of sickle hemoglobin gels: relevance to pathophysiology of sickling disorders.
The physical properties of hemoglobin (Hb) S gels formed under conditions relevant to in vivo conditions have been characterized using viscometry. Mixed Hb SF and Hb AS gels of hemoglobin concentrations and compositions found in patients with Hb S-HPFH (mild sickle-cell disease) and Hb AS (sickle-cell trait) have been compared to those of pure Hb S. The results may explain why red cells containing gels occlude the microcirculation and/or undergo hemolysis in patients with sickle-cell anemia, why patients with Hb S-HPFH are mildly affected with red-cell sickling, and why individuals with Hb AS have no clinical manifestations of the Hb abnormality.