Colorectal Cancer in Young African Americans: Is It Time to Revisit Guidelines and Prevention?
Previous studies have suggested an increase in the incidence of colorectal cancer (CRC) in young adults (younger than 50 years). Among older people, African Americans have disproportionally higher CRC incidence and mortality. We assessed whether this CRC disparity also applies to CRC diagnosed among young people. Using the Surveillance, Epidemiology, and End Results cancer registries, a population-based cancer registry covering 25.6 % of the United States' African American population, we identified patients diagnosed with CRC between the years of 2000-2012. The age-adjusted rates for non-Hispanic whites (NHW), African Americans, and Asian-Pacific Islanders (API) were calculated for the age categories 20-24, 25-29, 30-34, 35-39, and 40-44. CRC age-adjusted incidence is increasing among all three racial groups and was higher for African Americans compared to NHW and API across all years 2000-2012 (P < 0.001). Stage IV CRC was higher in African Americans compared with NHW, while there was higher stage III CRC in API compared with NHWs. CRC incidence is increasing among the young in all racial groups under study. This increase in frequency of CRC is true among young African American adults who display highly advanced tumors in comparison with other races. While the present attention to screening seems to have decreased CRC prevalence in individuals older than 50, special attention needs to be addressed to young African American adults as well, to counter the observed trend, as they have the highest incidence of CRC among young population groups by race/ethnicity.


Pseudoangiomatous stromal hyperplasia of the vulva presenting as a polypoid vulvar lesion: a lesion associated with anogenital mammary-like tissue-report of a case and review of the literature.
The study aimed to report the case of a patient with a polypoid vulvar lesion clinically thought to be an aggressive angiomyxoma. On examination, a 4.0 × 2.0 × 1.5-cm polypoid lesion was seen on the right labium majus. Histopathological findings of the excised mass were consistent with pseudoangiomatous stromal hyperplasia of the vulva Pseudoangiomatous stromal hyperplasia is a lesion of breast tissue but can occur in anogenital mammary-like glands. It must be distinguished from low-grade angiosarcoma.


S100B and response to treatment in major depression: a pilot study.
S100B is a protein which exerts both detrimental and neurotrophic effects, depending on its concentration in brain tissue. An increase of S100B in micromolar concentrations is observed in traumatic brain conditions and is associated with poor outcome. Micromolar levels of extracellular S100B in vitro may have deleterious effects. However, in nanomolar concentrations S100B has multiple neurotrophic effects in vitro may in vivo be regarded as a hallmark of neuroprotective efforts. This pilot study addresses the hypothesis that S100B serum concentrations may be of predictive validity for the response to antidepressant treatment in patients with major depression. S100B plasma levels were determined in 25 patients with major depression and 25 matched healthy controls using an immunofluorimetric sandwich assay. S100B plasma levels were significantly higher in major depressive patients than in healthy controls and positively correlated with treatment response after 4 weeks of treatment. In a linear regression model, a significant predictive effect was found only for S100B and severity of depressive symptoms upon admission. These results suggest that neuroprotective functions of S100B counterbalance neurodegenerative mechanisms that are involved in the pathophysiology of major depression and in the response to antidepressant treatment.


Suppressed cellular immunity in patients with nasopharyngeal carcinoma.
The subsets and functions of lymphocytes were investigated in patients with nasopharyngeal carcinoma (NPC). The patients were divided into two groups comprising tumor-bearing patients and those in remission. There was no difference in the proportion of T cells among tumor-bearing, remission and healthy control groups. The percentages of inducer/helper T cells and natural killer cells were smaller in the tumor-bearing group than in the control group whereas the percentage of suppressor T cells was greater in the tumor-bearing group. Phytohemagglutinin-stimulated blastogenesis was markedly suppressed in the tumor-bearing group. The responsiveness to interleukin-2 of blastogenesis and of natural killer and lymphokine-activated killer activities was lowered in the tumor-bearing group. These parameters in the remission group were intermediate between those of the tumor-bearing and control groups. These results suggest that cellular immunity is suppressed in patients with NPC and that the suppressed condition still remains even in remission. Immunotherapy is considered to be indispensable for the proper treatment of NPC.


Speech audiometry.
Two aspects of speech audiometry, namely, the speech reception threshold and the speech discrimination scores, have been discussed in the light of their development and present day administration. Evidence was presented to forego the concept of phonetic balance in discrimination test messages for the preferable attributes of familiarity and equated list difficulty. Campbell's redistribution of the W-22 word lists was proposed as an available solution to one of these parameters. Some delineation of the single "maximal" discrimination score's relation to the prediction of the hearing impaired individual's ability to understand continuous discourse was offered. Diagnostic implications of speech audiometry were not treated.


Georgia Blues launch new P4P program.
In Virginia, 41 hospitals are being awarded dollars 12 million for improvements in patient safety and health outcomes. The approach is tailored to each hospital, which can choose its own goals and objectives. Performance objectives are based on processes published by JCAHO, the Leapfrog Group, and other national organizations.


What to Expect From the Evolving Field of Geriatric Cardiology.
The population of older adults is expanding rapidly, and aging predisposes to cardiovascular disease. The principle of patient-centered care must respond to the preponderance of cardiac disease that now occurs in combination with the complexities of old age. Geriatric cardiology melds cardiovascular perspectives with multimorbidity, polypharmacy, frailty, cognitive decline, and other clinical, social, financial, and psychological dimensions of aging. Although some assume that a cardiologist may instinctively cultivate some of these skills over the course of a career, we assert that the volume and complexity of older cardiovascular patients in contemporary practice warrants a more direct approach to achieve suitable training and a more reliable process of care. We present a rationale and vision for geriatric cardiology as a melding of primary cardiovascular and geriatrics skills, thereby infusing cardiology practice with expanded proficiencies in diagnosis, risks, care coordination, communications, end-of-life, and other competences required to best manage older cardiovascular patients.


Systematic validation of an automated thalamic parcellation technique using anatomical data at 3T.
The thalamus is a brain region formed from functionally distinct nuclei, which contribute in important ways to various cognitive processes. Yet, much of the human neuroscience literature treats the thalamus as one homogeneous region, and consequently the unique contribution of specific nuclei to behaviour remains under-appreciated. This is likely due in part to the technical challenge of dissociating nuclei using conventional structural imaging approaches. Yet, multiple algorithms exist in the neuroimaging literature for the automated segmentation of thalamic nuclei. One recent approach developed by Iglesias and colleagues (2018) generates segmentations by applying a probabilistic atlas to subject-space anatomical images using the FreeSurfer software. Here, we systematically validate the efficacy of this segmentation approach in delineating thalamic nuclei using Human Connectome Project data. We provide several metrics quantifying the quality of segmentations relative to the Morel stereotaxic atlas, a widely accepted anatomical atlas based on cyto- and myeloarchitecture. The automated segmentation approach generated boundaries between the anterior, lateral, posterior, and medial divisions of the thalamus. Segmentation efficacy, as measured by metrics of dissimilarity (Average Hausdorff Distance) and overlap (DICE coefficient) within groups was mixed. Regions were better delineated in anterior, lateral and medial thalamus than the posterior thalamus, however all the volumes for all segmented nuclei were significantly different to the corresponding region of the Morel atlas. These mixed results suggest users should exercise care when using this approach to study the structural or functional relevance of a given thalamic nucleus.


No effect of cold pressor test-induced arousal on attentional benefits and costs in an endogenous spatial orienting paradigm.
Previous studies have shown that arousal can influence hemispatial bias, suggesting that changes in arousal affect the neural networks involved in spatial attention control. The goal of the present study was to measure the effects of increased arousal on endogenous attentional orienting. We used a Spatial Orienting Paradigm to quantify attentional benefits and costs as measures of attentional orienting and re-orienting responses and exposed participants (N = 25; Experiment 1) to a bilateral feet Cold Pressor Test (CPT) to manipulate arousal. Increases in subjective distress ratings and blood pressure confirmed the effect of CPT on arousal. Although no overall effects of CPT on reaction times in the Spatial Orienting Paradigm were detected, an exploratory analysis of sex-specific effects revealed a left-lateralised decrease in benefits and increase in costs after CPT exposure in the male subsample (N = 11). To confirm these preliminary results, we repeated the experiment in a larger sample (N = 29, all male), but found no effect of CPT on orienting, with moderate to strong evidence in favour of a model excluding all (interaction) effects of CPT exposure (all BFIncl < 0.3). Instead, our replicated results indicate that voluntary orienting is unaffected by CPT-induced increases of arousal. In the light of previous studies, and keeping in mind the interpretative challenges of null results, we discuss how and why our findings may be specific to endogenous as opposed to exogenous orienting and how arousal could possibly lead to the previously established effects on visuospatial bias without simultaneously affecting orienting and the underlying attention control networks.


Nanomedicine applied to translational oncology: A future perspective on cancer treatment.
The high global incidence of cancer is associated with high rates of mortality and morbidity worldwide. By taking advantage of the properties of matter at the nanoscale, nanomedicine promises to develop innovative drugs with greater efficacy and less side effects than standard therapies. Here, we discuss both clinically available anti-cancer nanomedicines and those en route to future clinical application. The properties, therapeutic value, advantages and limitations of these nanomedicine products are highlighted, with a focus on their increased performance versus conventional molecular anticancer therapies. The main regulatory challenges toward the translation of innovative, clinically effective nanotherapeutics are discussed, with a view to improving current approaches to the clinical management of cancer. Ultimately, it becomes clear that the critical steps for clinical translation of nanotherapeutics require further interdisciplinary and international effort, where the whole stakeholder community is involved from bench to bedside. From the Clinical Editor: Cancer is a leading cause of mortality worldwide and finding a cure remains the holy-grail for many researchers and clinicians. The advance in nanotechnology has enabled novel strategies to develop in terms of cancer diagnosis and therapy. In this concise review article, the authors described current capabilities in this field and outlined comparisons with existing drugs. The difficulties in bringing new drugs to the clinics were also discussed.


One-hour plasma glucose combined with skin autofluorescence identifies subjects with pre-diabetes: the DIAPASON study.
The major challenge for diabetes prevention is early identification of individuals at risk to allow for implementation of measures to delay the onset of future disease. Measures such as fasting plasma glucose (FPG), 2-hour plasma glucose (2hPG), and glycosylated hemoglobin (HbA1c) are equally appropriate for identifying pre-diabetes and diabetes, but do not all identify the disease in the same individual. We tested the utility of a diagnostic method combining FPG, 2hPG and HbA1c for early evaluation and easy identification of pre-diabetes. 531 subjects underwent skin autofluorescence (SAF) and glycemia analyses. We created two classification groups based on the American Diabetes Association diagnosis guidelines: (1) based on 2hPG and (2) based on a new combination of three glycemia parameters (the three-criteria strategy (3-c)). Logistic regression modeling was used to estimate the associations. SAF showed high associations for both 3-c definition and 2hPG definition alone. These associations appeared stronger in 3-c than those in 2hPG. The non-invasive SAF measurement outperformed 2hPG in the detection of dysglycemia or pre-diabetes. Stepwise selections identified 1-hour postload glucose (1hPG) as variable identifying pre-diabetes using the 2hPG criterion, and the model based on 1hPG plus SAF appeared to be the best association using the 3-c strategy. 1hPG coupled with SAF showed a strong association in the evaluation of pre-diabetes using the 3-c method.


Evaluating new candidate SNPs as low penetrance risk factors in sporadic breast cancer: a two-stage Spanish case-control study.
A polygenic model has been proposed in order to explain the genetic susceptibility to sporadic breast cancer. According to this model, common population variants would be responsible for low to modest effects on the risk of developing the disease. We have carried out a high-throughput SNP genotyping project in order to shed some light on the complex genetic aetiology of non-familial breast cancer. Ninety-one genes have been selected because of their implications in several candidate cell pathways for breast cancer. A total of 640 SNPs in these genes were genotyped in a series of 450 consecutive cases and 448 controls from mainland Spain. Promising SNPs were then studied in an independent series of 294 cases and 299 controls from the Canary Islands. In the first case-control series we identified 25 SNPs with P-values below 0.05 (under a 1 df Chi-square test), five of them with P-values below 0.01 (best=0.0008). In the stage 2 Canary Islands series, odd ratios (OR) for two SNPs in HUS1 were in a consistent direction. SNPs located at the gene HUS1 are good candidates for further investigation in independent association studies and functional assays.


Self-reported cannabis use is not associated with greater opioid use in elective hand surgery patients.
The purpose of this study was to examine the influence of preoperative marijuana use on postoperative opioid use during the first three postoperative days (POD 1-3) after surgery, and on expectations of pain control, resiliency, and quality-of-life scores. All patients presenting to a single institution undergoing elective hand or upper extremity outpatient surgery were asked to complete pre- and postoperative questionnaires. Preoperative questionnaires collected information on demographics, marijuana use, tobacco use, procedure type, self-assessed health, pain control expectations, and EuroQol-5D (EQ-5D) scores. At the first postoperative visit, patients self-reported opioid consumption from POD 1-3. Multivariate linear regression analysis was used to determine which patient characteristics were predictive of greater postoperative opioid consumption during POD 1-3. Self-reported marijuana users were younger, less healthy, and more likely to use tobacco compared to non-users. Marijuana users and non-users were comparable in their use of pain medication (including non-opioids), rates of chronic pain diagnoses, and self-reported pain tolerance. EQ-5D scores were lower in marijuana users than non-users (0.64 vs. 0.72). Marijuana users and non-users were prescribed comparable quantities of opioids during the first 14 days after surgery (176 ± 148 vs 115 ± 87). Multiple linear regression analysis revealed that lower preoperative EQ-5D scores, rather than marijuana use, were associated with increased opioid consumption during POD 1-3. Preoperative marijuana use was not independently associated with increased opioid use during POD 1-3 after elective hand and upper extremity surgery; instead, an association with lower preoperative EQ-5D scores was identified. II, prospective cohort study.


Proteomics analysis reveals several metabolic alterations in cyanobacterium Anabaena sp. NC-K1 in response to alpha-cypermethrin exposure.
In the current study, the effect of the EC50 and LC90 concentrations of pyrethroid insecticide alpha-cypermethrin to cyanobacteria Anabaena sp. NC-K1 was investigated at different time exposures (1st day, 4th day and 7th day) with reference to growth, photosynthetic pigments, oxidative damage and antioxidant defence system. Superoxide dismutase (1.38-fold), peroxidase (5.04) and proline content (2.27-fold) were enhanced compared to the control. After performing 2D gel electrophoresis at 1st day EC50 exposure, where appropriate differences in the biochemical and physiological parameters were observed, 22 differentially accumulated proteins (20 upregulated and 2 downregulated) were selected for mass spectrometry. Out of 42 proteins identified, 20 upregulated protein spots were classified into twelve categories according to their metabolic functions. Proteins related to photosynthesis (phycobilisome rod-core linker polypeptide, rubisco), stress responses (Hsp70, Hsp40, catalase family peroxidase), translation (elongation factor Tu) and amino acid biosynthesis and metabolism (3-phosphoshikimate 1-carboxyvinyl transferase) were significantly upregulated. Additionally, proteins involved in transcription and DNA repair (Snf-2 histone linker phd ring helicase, RNA polymerase sigma factor RpoD and Holliday junction ATP-dependent DNA helicase RuvA) were considerably upregulated. Upregulation of these proteins against pesticide stress presumably maintained the photosynthesis, energy metabolism, carbohydrate metabolism, transport and signalling proteins, transcription, translation and DNA repair. Additionally, these proteins might involve in sufficient detoxification of ROS and play a crucial role in damage removal and repair of oxidized proteins, lipids and nucleic acids. Taken together, Anabaena sp. NC-K1 responded towards alpha-cypermethrin stress via modulating its proteome to maintain its cellular metabolism and homeostasis.


Comparison of the in vitro dissolution properties and in vivo steady-state pharmacokinetics of two sustained-release theophylline preparations.
The sustained-release properties and relative bioavailability of Theolin Retard and Pharphylline Retard were studied in eight healthy adults after treatment for five days with twice daily 450 mg, respectively 425 mg. During the day-time dosing interval on the fourth and fifth day theophylline plasma concentrations were assayed by HPLC. After intake of Theolin Retard, minimum theophylline plasma concentrations were significantly higher, fluctuations in theophylline plasma concentrations were significantly smaller and t75 (the period within a dosing interval during which the plasma concentration exceeds 75% of the maximal concentration) was significantly longer than after Pharphylline Retard. Maximal concentrations and AUC values were not significantly different. For both products the plasma concentration time-curves on day 5 were significantly lower than on day 4. In vitro dissolution tests confirmed the more sustained release of theophylline from Theolin Retard. These results indicate an equal extent of absorption from the two products but better sustained-release properties for Theolin Retard.


Curcumin and tumor immune-editing: resurrecting the immune system.
Curcumin has long been known to posses medicinal properties and recent scientific studies have shown its efficacy in treating cancer. Curcumin is now considered to be a promising anti-cancer agent and studies continue on its molecular mechanism of action. Curcumin has been shown to act in a multi-faceted manner by targeting the classical hallmarks of cancer like sustained proliferation, evasion of apoptosis, sustained angiogenesis, insensitivity to growth inhibitors, tissue invasion and metastasis etc. However, one of the emerging hallmarks of cancer is the avoidance of immune system by tumors. Growing tumors adopt several strategies to escape immune surveillance and successfully develop in the body. In this review we highlight the recent studies that show that curcumin also targets this process and helps restore the immune activity against cancer. Curcumin mediates several processes like restoration of CD4(+)/CD8(+) T cell populations, reversal of type-2 cytokine bias, reduction of Treg cell population and suppression of T cell apoptosis; all these help to resurrect tumor immune surveillance that leads to tumor regression. Thus interaction of curcumin with the immune system is also an important feature of its multi-faceted modes of action against cancer. Finally, we also point out the drawbacks of and difficulties in curcumin administration and indicate the use of nano-formulations of curcumin for better therapeutic efficacy.


Impact of COVID-19 Pandemic on Non-COVID-19 Maternal Mortalities in a Tertiary Health Care Center of North India.
While dealing with the COVID-19-related morbidities and mortalities in general, its adverse impact on non-COVID-19 maternal mortalities was completely forgotten so our aim is to study the adverse consequences of the COVID-19 pandemic on non-COVID-19 hospital births and non-COVID-19 maternal mortalities. Retrospective observational study was done in the Department of Obstetrics and Gynecology, Swaroop Rani Hospital, Prayagraj, to compare the non-COVID-19 hospital births, referrals, and non-COVID-19 maternal mortalities during 15 months of pre-pandemic period (March 2018 to May 2019) and 15 months of pandemic period (March 2020 to May 2021 period) and to assess their relation to GRSI using chi-square test, paired T test and Pearson's Correlation Coefficient. The total non-COVID-19 hospital births decreased by 4.32% in pandemic period as compared to pre-pandemic period. Monthly hospital births decreased drastically, to 32.7% during the end of the first pandemic wave and to 60.17% during the second wave. 67% increase in the total referral and a significant decrease in the quality of referral leading to significantly higher non-COVID-19 maternal mortality figures (p value 0.00003) during the pandemic period. Leading causes of mortalities were uterine rupture (p value 0.00001), septic abortion (p value 0.0001), primary postpartum hemorrhage (p value 0.002) and preeclampsia (p value 0.003). While the world is talking only about COVID deaths, increased non-COVID-19 maternal mortalities during the COVID pandemic need equal attention and call for more stringent government guidelines for the care of non-COVID-19 pregnant women as well during the pandemic period.


Molecular parameters of semisynthetic derivatives of gangliosides and sphingosine in monolayers at the air-water interface.
The molecular parameters (molecular area, surface potential, collapse pressure, dipole moment contributions) of semisynthetic derivatives of ganglioside GM1 and of sphingosine were studied in lipid monolayers at the air-NaCl (145 mM, pH 5.6) interface at 22 +/- 0.3 degrees C. The chemical modifications included alterations of the fatty acyl chain moiety linked to the 2-amino position of the sphingosine (Sph) base. The compounds studied were PKS-1 (N-acetyl Sph), PKS-2 (N-chloroacetyl Sph), PKS-3 (N-dichloroacetyl Sph), PKS-4 (N-trichloroacetyl Sph), Lyso-GM1 (ganglioside GM1 lacking the N-linked fatty acyl chain and the N-acetyl group on the sialic acid), Liga-4 (N-acetyl, lyso[NeuAc]GM1) and Liga-20 (N-dichloroacetyl, lyso[NeuAc]GM1). Relatively small modifications of the chemical structure of sphingolipids introduce dramatic consequences on their surface molecular properties. The absence of the long chain fatty acyl moiety and of the N-acetyl group on the neuraminic acid in Lyso-GM1 leads to a more condensed behavior and to an increase of the collapse pressure compared with GM1. The acetylation or chloroacetylation at the 2-amino position in Liga-4 and Liga-20 induce an expansion of the surface pressure-area isotherm and a decrease of the collapse pressure. The limiting molecular areas of GM1 derivatives, taken at the collapse pressure point, are consistent with the oligosaccharide chain being oriented approximately perpendicularly to the interface. Sphingosine shows a liquid expanded isotherm. The acetylation and successive chlorination of the acetyl residue at the 2-amino position of Sph cause a progressive increase in the limiting molecular area. The variation of the resultant dipole moment under compression, calculated from the surface potential values, suggests the reorientation of selective groups within these molecules that depend on the degree of intermolecular packing. Thermodynamic-geometric correlations on the basis of the molecular parameters of these derivatives suggest that small alterations of the substituent group at the 2-amino position of Sph could have large and amplified consequences on the type, curvature and stability of the possible self-aggregated structure that these lipids may form in aqueous medium.


Reconstitution of human lymphocytes following ex vivo expansion of human umbilical cord blood CD34+ cells and transplantation in Rag2-/- gamma c-/- mice model.
Due to ethical issues, in vivo studies of the human immune system have been difficult. Thus, small-animal xenotransplantation models have been employed, although they scarcely sustain a human immune response. In this study, we compared human cell repopulation tendencies and functionality in Rag2-/- gamma c-/- mice following various ex vivo expanded human hematopoietic stem cells (HSCs). Human umbilical cord blood (UCB) CD34+ cells were cultured for 7 days with a cytokine combination of stem cell factor, Flk2/Flt3 ligand, and thrombopoietin, with absence or presence of rhIL-3, then transplanted into Rag2-/- gamma c-/- mice. Reconstituted human lymphocytes were analyzed based on the expression of CD45 as well as CD3, CD19, and CD56 in peripheral blood (PB) until 16 weeks after transplantation. BrdU assay and functional analysis of reconstituted human lymphocytes used PHA- or rhIL-2-stimulated splenocytes and bone marrow cells from recipient mice. The percentage of human CD45dim cells, not CD45bright cells, in PB of mice transplanted with cultured HSCs with rhIL-3 was much higher than in the group without rhIL-3 (approximately 2.5-fold at week 10 posttransplantation). The humanized mice showed systemic repopulation with a comprehensive array of human lymphohematopoietic cells, including T, B, natural killer (NK) cells, and even dendritic cells. However, the expression level was also dim. The number of CD3+ T cells and CD56+ NK cells was especially increased in the presence of rhIL-3. In addition, after in vitro restimulation proliferation assays and NK activity of interferon-gamma secretion showed greater effects in the presence of rhIL-3. These data suggested that the development of a diverse repopulation of human lymphocytes was possible in Rag2-/- gamma c-/- mice after transplantation of cultured UCB CD34+ HSCs with interleukin-3.


Photometric oculometry: a slit-lamp technique for determination of intraocular distances.
The optics of slit-lamp fluorophotometry were analyzed. This analysis forms the basis of the formulation of a slit-lamp technique called photometric oculometry, which makes it possible to measure intraocular distances. A comparison between ultrasonography and photometric oculometry in the determination of the ocular axial length was carried out. Measurements in 11 eyes showed statistical agreement, with a mean of 22.90 mm, SD +/- 1.815 (ultrasound), and a mean of 23.17 mm, SD +/- 1.667 (photometry). The principle involved in photometric oculometry is of special importance for the calculation of the blood-retinal barrier permeability to fluorescein by vitreous fluorophotometry, since the method allows the volume of the vitreous body to be estimated directly from the slit-lamp fluorophotometer recording.


Importance of serum cholesterol level in development of diabetic autonomic neuropathy.
Changes of cardiac beat-to-beat variation (BBV)--an index of autonomic nerve function--were examined on 2 occasions in an interval of 3.7-5.9 years in 33 diabetics. A total of 204 healthy people, ranging from 11 to 88 years of age, were also subjected to the BBV measurement as controls. In diabetics, a lower rate of change of BBV than the control value was considered as deterioration. Glycohemoglobin was measured every month. The mean of serum cholesterol on the 2 occasions nearest to each time BBV examination was taken for the evaluation of cholesterol level. No significant difference was observed in the control state of blood glucose in terms of glycohemoglobin between the groups with and without BBV deterioration. In 20 subjects with normal cholesterol level, 11 showed a BBV deterioration, and in 13 subjects with abnormal cholesterol level 12 deteriorated in BBV. Therefore, diabetics with normal serum cholesterol level did not seem to exacerbate autonomic neuropathy in terms of BBV.


Pharmacogenetics-based therapeutic recommendations--ready for clinical practice?
Although considerable progress has been made in basic pharmacogenetic research, less has been demonstrated in the application of pharmacogenetics (PGx)-based diagnostics to drug development and in clinical practice. There are drugs that are currently used in the clinic for which individualized therapy could be beneficial based on PGx data. However, specific, actionable recommendations on how to implement individualized therapy--particularly with respect to dosage--still have to be developed. Moreover, to apply PGx efficiently in clinical drug development, and later in drug therapy, study designs and the generation and handling of PGx data need to become more standardized. Here, we argue for the development of concise guidelines for implementation of PGx analyses in drug development and therapy.


Visualization of Live Cochlear Stereocilia at a Nanoscale Resolution Using Hopping Probe Ion Conductance Microscopy.
The mechanosensory apparatus that detects sound-induced vibrations in the cochlea is located on the apex of the auditory sensory hair cells and it is made up of actin-filled projections, called stereocilia. In young rodents, stereocilia bundles of auditory hair cells consist of 3-4 rows of stereocilia of decreasing height and varying thickness. Morphological studies of the auditory stereocilia bundles in live hair cells have been challenging because the diameter of each stereocilium is near or below the resolution limit of optical microscopy. In theory, scanning probe microscopy techniques, such as atomic force microscopy, could visualize the surface of a living cell at a nanoscale resolution. However, their implementations for hair cell imaging have been largely unsuccessful because the probe usually damages the bundle and disrupts the bundle cohesiveness during imaging. We overcome these limitations by using hopping probe ion conductance microscopy (HPICM), a non-contact scanning probe technique that is ideally suited for the imaging of live cells with a complex topography. Organ of Corti explants are placed in a physiological solution and then a glass nanopipette-which is connected to a 3D-positioning piezoelectric system and to a patch clamp amplifier-is used to scan the surface of the live hair cells at nanometer resolution without ever touching the cell surface.Here, we provide a detailed protocol for the imaging of mouse or rat stereocilia bundles in live auditory hair cells using HPICM. We provide information about the fabrication of the nanopipettes, the calibration of the HPICM setup, the parameters we have optimized for the imaging of live stereocilia bundles and, lastly, a few basic image post-processing manipulations.


Synthesis and antiviral activity of 2'-fluorohexopyranosyl nucleosides.
2'-Fluorohexopyranosyl nucleosides 1a and 1b which contained a bioisosteric double bond and a fluorine were synthesized in 12 steps, starting from D-galactose. During diethylaminosulfur trifluoride (DAST) fluorination, retention of stereochemistry was observed through the participation of methoxy or chloro group at the 6-position of the purine base. The final nucleosides 1a and 1b were found to be inactive against HIV-1 and HSV-1,2.


Effect of Linguistic and Musical Experience on Distributional Learning of Nonnative Lexical Tones.
Evidence suggests that extensive experience with lexical tones or musical training provides an advantage in perceiving nonnative lexical tones. This investigation concerns whether such an advantage is evident in learning nonnative lexical tones based on the distributional structure of the input. Using an established protocol, distributional learning of lexical tones was investigated with tone language (Mandarin) listeners with no musical training (Experiment 1) and nontone language (Australian English) listeners with musical training (Experiment 2). Within each experiment, participants were trained on a bimodal (2-peak) or a unimodal (single peak) distribution along a continuum spanning a Thai lexical tone minimal pair. Discrimination performance on the target minimal pair was assessed before and after training. Mandarin nonmusicians exhibited clear distributional learning (listeners in the bimodal, but not those in the unimodal condition, improved significantly as a function of training), whereas Australian English musicians did not (listeners in both the bimodal and unimodal conditions improved as a function of training). Our findings suggest that veridical perception of lexical tones is not sufficient for distributional learning of nonnative lexical tones to occur. Rather, distributional learning appears to be modulated by domain-specific pitch experience and is constrained possibly by top-down interference.


Ethanol effects on active Na+ and K+ transport in cultured fetal rat hepatocytes.
To define further the influence of ethanol on membranes, its effects on Na+ pump function were studied in monolayer cultures of fetal rat hepatocytes. The effects of ethanol (2 and 4 mg/ml) on total K+ influx, ouabain-sensitive K+ influx, Na+ pump density (from specific [3H]ouabain binding), pump turnover rates and intracellular Na+ were measured following exposure of the cells to ethanol for 1-24 hr. In parallel studies, the effects of ethanol (2 mg/ml) on cell water content and membrane fluidity were measured. Ethanol had no immediate effect on K+ influx, but after 1 hr ethanol in concentrations of 2 and 4 mg/ml decreased the total K+ influx (mumol/10(11) cells/sec) from a control of 8.5 +/- 0.64 to 4.46 +/- 0.50 and 4.09 +/- 0.26 respectively (N = 6 for each experiment; P less than 0.001). This represented the maximum effect of ethanol since after 6 and 24 hr of ethanol treatment the K+ influx had increased towards control levels but remained significantly (P less than 0.01 for 2 mg/ml and P less than 0.001 for 4 mg/ml) below that in control cells even at 24 hr. The decrease in K+ influx reflected a decrease in mean ouabain-sensitive K+ influx from a control of 5.87 to 3.24 and 2.70 (mumol/10(11) cells/sec) after a 1-hr treatment with 2 and 4 mg ethanol/ml medium respectively. Ethanol (2 mg/ml) treatment for 1-hr decreased Na+ pump density (x 10(5) molecules ouabain per cell) from a control of 2.80 +/- 0.30 to 1.70 +/- 0.11 (P less than 0.001). At 6 and 24 hr [3H]ouabain binding showed a pattern similar to that seen with the K+ influx, tending to return to pretreatment levels. There was no change in individual pump turnover rates in the presence of ethanol. Following exposure to ethanol, cellular Na+ content steadily increased over the first 6 hr and then returned to control levels. When corrected for parallel changes in cell volume, however, intracellular Na+ concentration increased by 17% (P less than 0.01) after 1 hr and thereafter remained at this higher level throughout the 24-hr period. Measurements of membrane fluidity showed that it was increased markedly by ethanol at a concentration of 2 mg/ml and that the effect bore a close temporal relationship to the changes in active K+ influx and Na+ pump density. We conclude that ethanol has a depressant effect on hepatic Na+ pump function, resulting in an increase in intracellular Na+ and an eventual gain in cell water.(ABSTRACT TRUNCATED AT 400 WORDS)


Determination of direct alcohol markers: a review.
Alcohol is the most popular legal drug used in our society today, and its consumption by pregnant women remains an important public health problem. Gestational alcohol consumption can result in a continuum of adverse fetal outcomes known as fetal alcohol spectrum disorder (FASD). Effective strategies are needed to prevent the increasing adoption of risky drinking behaviors. Because ethanol itself is only measurable for a few hours after ethanol intake in conventional matrices including blood, urine, and sweat, these matrices are only useful to detect recent ethanol exposure. Since approximately early 2000, the non-oxidative ethanol metabolites have received increasing attention because of their specificity and, in some cases, wide time window of detection in non-conventional matrices including hair and meconium. In the attempt to update analytical methods for the determination of non-oxidative markers of alcohol, the objective of this study is to review published studies that measure fatty-acid ethyl esters (FAEE), ethyl glucuronide (EtG), and phosphatidylethanol (PEth) in alternative biological matrices, focusing on the extraction and detection methods and full analytical conditions used.


The equivalent refractive index of the crystalline lens in childhood.
Despite the importance of crystalline lens power in ocular development, schematic refractive index values used to calculate lens power have been validated for children. We measured refractive error and ocular component dimensions in 519 schoolchildren, calculating lens power using phakometrically measured lens radii and three different refractive index profiles: (1) Gullstrand-Emsley schematic indices [Gullstrand-Emsley lens power (GELP)]; (2) a 10-shell gradient index model [gradient index lens power (GILP)]; and (3) the equivalent refractive index (IND) needed to bring calculated and measured refractive error into agreement [calculated lens power (CLP)]. GELP was significantly lower than either GILP or CLP, indicating the Gullstrand-Emsley refractive index of 1.416 is too low for use in children. Variation in IND cannot be explained by measurement error alone. GILP and CLP also differed as a function of lens shape, with GILP greater than CLP at steeper external curvatures and less than CLP at flatter external curvatures. Variation in equatorial gradient index profile as a function of lens shape is proposed as an explanation for this bias. Equivalent index appears to be a useful tool for encompassing individual variation in lens gradient profiles as well as for assessing the relative role of lens surface curvature and refractive index changes during lens power development in childhood.


[Deficiency of the 6th component of complement. Recurrent meningococcal and streptococcal meningitis].
A 39-year old male patient with a history of four episodes of meningitis (two of them due to Neisseria meningitidis) since the age of 12 developed meningitis caused by an ungroupable streptococcus. Deficiency of the sixth component of complement was discovered, making this the ninth case of recurrent meningitis associated with C6 deficiency. Streptococcal meningitis had never been reported in such cases.


Alterations in redox homeostasis and prostaglandins impair endothelial-dependent vasodilation in euglycemic autoimmune nonobese diabetic mice.
We report herein the novel observation that alterations in oxidant/antioxidant balance are evident and cause vascular dysfunction in aortae of prediabetic nonobese-diabetic mice (NOD). We found that nitrotyrosine, a biochemical marker of oxidant stress, was higher in the NOD aortae when compared to age-matched non-autoimmune BALB/c controls or the diabetes-resistant NOD congenic strain, NOD.Lc7. The oxidant stress was localized to the intimal and medial layers, and endothelium-dependent relaxation to acetylcholine was decreased in isolated aortic rings from NOD mice. Inhibition of nitric oxide synthesis caused an endothelium-dependent contraction, and treatment with either a selective thromboxane A2/prostaglandin H2 receptor antagonist or a non-isozyme-specific cyclooxygenase inhibitor reversed this effect. Aortic rings from NOD.Lc7 did not display the paradoxical vasoconstriction. Furthermore, the vascular dysfunction was caused by oxidative stress, as treatment with a superoxide dismutase mimetic in vivo or with native antioxidant enzymes ex vivo inhibited the tissue oxidant stress and restored endothelium-dependent relaxation. Endothelial function was also restored by the inhibitors of NAD(P)H oxidase, diphenylene iodonium or apocynin. Our studies indicate that an oxidant stress that occurs prior to the onset of diabetes in this mouse model contributes to endothelial dysfunction independently of overt diabetes.


Salvage radiotherapy after robot-assisted laparoscopic radical prostatectomy.
To assess the incidence and efficacy of salvage radiotherapy (SRT) after robot-assisted radical prostatectomy (RARP). RARP has been linked to an increased use of adjuvant treatments. If RARP would result in an increased local recurrence rate, response rates to SRT could be expected to be better after RARP than after more conventional methods of prostatectomy. The incidence and efficacy of SRT in a RARP population were compared with nomogram prediction. Patient data were prospectively registered. Biochemical recurrence (BCR) was defined as a prostate-specific antigen (PSA) ≥0.1 ng/mL. SRT was offered to men with BCR after RARP. The Stephenson nomogram predictions were compared with outcome after SRT. Of 1087 men, 157 (14.4%) received SRT for BCR or persistent PSA levels during a median follow-up of 1078 days after RARP. Median PSA level before SRT was 0.2 ng/mL. Three-year BCR-free rate was 64% for men after SRT. pN (pNx, pN0, pN1) and PSA level before SRT were independent predictors of the BCR interval after SRT. Men with more extensive fascia preservation were more likely to respond favorably to SRT. The Stephenson nomogram prediction showed a concordance rate of 0.66 in this RARP population. Limitations of the study are the retrospective design and limited follow-up duration. In our RARP series, the use and efficacy of SRT were comparable with open prostatectomy series. The Stephenson nomogram reliably predicted outcome in patients with RARP with SRT, suggesting that similar characteristics predict response to SRT after RARP compared with open prostatectomy.


Anticardiolipin antibodies in homosexual men: prevalence and lack of association with human immunodeficiency virus (HIV) infection.
Tests for anticardiolipin antibodies (ACL) on sera from 100 male homosexuals and 60 male heterosexuals showed that 57% of the homosexuals, in contrast to none of the heterosexuals, were ACL positive. The ACL were predominantly of the immunoglobulin G isotype and, in a high proportion of cases tested (82%), were reactive with other phospholipids, especially those with a negative charge. ACL were not related to the clinical status of the homosexuals, being evenly distributed among 40 with acquired immunodeficiency syndrome (AIDS), 20 with AIDS-related complex (ARC), 20 with asymptomatic infection with human immunodeficiency virus (HIV) and/or lymphadenopathy syndrome, and 20 who were HIV-antibody negative. Nor were they associated with thrombocytopenia, thrombosis, neurologic disease, a biological false-positive test for syphilis (BFP), or antibodies to DNA. It is concluded that factors other than infection with HIV are responsible for ACL positivity in homosexual males and that the epitopes recognized by ACL in this group are distinct from those associated with thromboembolism or the BFP reaction or cross-reactive with DNA.


A comparison of BOX-PCR and pulsed-field gel electrophoresis to determine genetic relatedness of enterococci from different environments.
Genetic relatedness of enterococci from poultry litter to enterococci from nearby surface water and groundwater in the Lower Fraser Valley regions of British Columbia, Canada was determined. A new automated BOX-PCR and Pulsed-Field Gel Electrophoresis (PFGE) were used to subtype enterococcal isolates from broiler and layer litter and surface and groundwater. All surface water samples (n = 12) were positive for enterococci, as were 11% (3/28) of groundwater samples. Enterococcus faecium (n = 90) was isolated from all sources, while Enterococcus faecalis (n = 59) was isolated from all sources except layer litter. The majority of E. faecalis originated from broiler litter (28/59; 47.5%) while the majority of E. faecium were isolated from layer litter (29/90; 32.2%). E. faecalis grouped primarily by source using BOX-PCR. Isolates from water samples were dispersed more frequently among PFGE groups containing isolates from poultry litter. E. faecium strains were genetically diverse as overall clustering was independent of source by both molecular methods. Subgroups of E. faecium isolates based upon source (layer litter) were present in BOX-PCR groups. Three individual E. faecalis groups and two individual E. faecium groups were 100% similar using BOX-PCR; only one instance of 100% similarity among isolates using PFGE was observed. Although enterococci from litter and water sources were grouped together using BOX-PCR and PFGE, isolates originating from water could not be definitively identified as originating from poultry litter. Automation of BOX-PCR amplicon separation and visualization increased the reproducibility and standardization of subtyping using this procedure.


Emerging treatments for itch in atopic dermatitis: A review.
Atopic dermatitis is a chronic inflammatory skin condition with a high prevalence that is increasing. The most universal symptom in patients with atopic dermatitis is pruritus; it is often the most troublesome symptom. New insights on the mechanism of itch in patients with eczema have been elucidated, involving cross-talk between neural and immune systems, which have advanced our treatments significantly. In the last few years, there are emerging treatments currently undergoing investigation that yield a promising outlook in treating this symptom. In this review, we aimed to provide an updated overview of future treatments undergoing phase II and III clinical trials that may be used to treat pruritus of atopic dermatitis.


Infrequent point mutations in codons 12 and 61 of ras oncogenes in human hepatocellular carcinomas.
DNA from human hepatocellular carcinomas (HCC) were analysed for the presence of mutations in codons 12 and 61 of the K-ras, H-ras and N-ras genes. The relevant ras sequences were amplified in vitro using the polymerase chain reaction and point mutations detected by selective hybridisation using mutation-specific synthetic oligonucleotides. In one of the 19 HCCs a mutation in codon 61 of the K-ras gene was detected, whilst in 3/19 HCCs a mutation was found in codon 61 of the N-ras gene. The mutations were all heterozygous A-T transversions and were found in HCCs arising in patients with underlying cirrhosis. In two of these patients where the corresponding normal tissue was available only the wild-type ras gene was detected, indicating that oncogenic activation of the ras gene was a consequence of somatic mutation. In another patient the same mutation in codon 61 of the N-ras gene was found in cirrhotic liver tissue and in all four patients the same mutation was also detected in formalin-fixed, paraffin-embedded liver biopsy HCC tissue obtained at diagnosis. These results indicate that mutational activation of the ras genes at codon 61 is an infrequent but possibly early event in the development of HCC in Britain.


[The cytocentrifuge in cerebrospinal fluid cytodiagnosis].
Cytocentrifugation enables high-quality liquor preparations rich in cells to be obtained, offering certain advantages over Sayk's sediment chamber method.


The effect of myo-inositol deficiency on lipid metabolism in rats. III. The mechanism of an enhancement in lipolysis due to myo-inositol deficiency in rats.
myo-Inositol deficiency in rats produced an overaccumulation of triacylglycerols in the liver due to stimulated lipolysis in the adipose tissue (Hayashi, E., Maeda, T. and Tomita, T. (1974) Biochim. Biophys. Acta 360, 134--155). The mechanism of the enhancement in lipolysis has now been investigated. The lipolytic response to adrenalin, corticotropin and insulin of the epididymal adipose tissue did not change due to the deficiency, but hormone-sensitive lipase activity, plasma adrenalin level and blood pressure were higher in the deficient rats. Adrenalectomy had no influence, but administration of sympathetic nervous blockers (reserpine, hexamethonium and bupranolol) inhibited the liver lipid deposition and an increase of serum free fatty acids in the deficient rats. These results indicate that the enhancement in lipolysis is mediated by an excitation of sympathetic nerve terminals innervating in the adipose tissues.


Genetic manipulation of two fowlpox virus late transcriptional regulatory elements influences their ability to direct expression of foreign genes.
Fowlpox virus (FWPV) is currently used as a vector to express foreign genes of various poultry and mammalian pathogens. However, due to limited information available about the primary structure of FWPV promoters required for an optimal transcriptional efficiency, the full potential of FWPV as an expression vector has not been completely realized. To dissect such transcriptional regulatory elements at the molecular level, we selected two FWPV promoters dictating contrasting levels of expression of acidic-type inclusion body protein gene (P190) and A15L vaccinia virus homolog of FWPV (P180) for site-directed mutagenesis studies. The transcriptional activity of mutated promoters was analyzed based on their ability to transcribe a reporter gene, lacZ, and translation of the resultant mRNA into functional protein. Replacement of the spacer sequences of P180 with those of P190 resulted in a five-fold increase in mRNA and a 17.6-fold increase in protein over those with its parental promoter, P180. Similarly, replacement of a thymidine after the start codon with guanosine resulted in a 2.3-fold increase in lacZ mRNA and a seven-fold increase in protein. Combining these substitutions in P180SG produced a maximum increase in mRNA and protein of 6.7- and 29.9-fold, respectively, over concentrations with its parental P180 promoter. The promoter activity of P180SG was comparable to that of the strongest natural promoter, P190. The amount of protein per transcript generated by the mutated promoters of P180 increased to at least three times that with the parental P180. In contrast, similar replacements in P190 resulted in a 40-50% reduction in mRNA and protein in all the mutated promoters. We discuss the significance of spacer sequence and the purine after the start codon in the context of a high level of expression.


Effects of a lesion in the dorsal raphe nuclei performed during the juvenile period of the female rat, on puberty.
The present study examined the effect of thermic lesions on the dorsal raphe nuclei (DRN) of the female rat, performed at various ages during the prepubertal period (21, 24, or 27 days), on puberty, and at first ovulation. In comparison with sham-operated animals, the age of vaginal opening and first vaginal oestrus was delayed in rats with a DRN lesion performed at the end of the infantile period (day 21) (45.6+/-0.94 vs. 89.9+/-1.03, p < 0.05), whereas differences were not observed in animals with lesions performed during the juvenile period (day 24 or 27). The number of ova shed by ovulating animals was greater in those rats with lesions performed on day 24 or 27 (9.7+/-0.4 vs. 7.4+/-0.4; 9.5+/-0.5 vs. 7.7+/-0.4, p < 0.05). Ovarian follicular atresia in these animals was significantly lower than in control and sham-operated ones. On the day of first vaginal oestrus and 48 h after the lesion, the serotonin-hypothalamic concentration decreased in all lesioned animals. Present results support the idea of the participation of the serotoninergic system, arising from the DRN, in the neuroendocrine mechanisms regulating the onset of puberty and the first ovulation, with variations depending on animal maturity.


A new approach to separation of voltage-activated Ca currents in rat melanotrophs.
The whole cell patch clamp technique was used to study voltage-activated calcium currents in rat melanotrophs. Voltage ramps of the same amplitude and of different durations were applied to separate Ca currents with different electrophysiological properties. The application of those with voltage gradient of 0.5 V/s resulted in a clear separation of two inward current components, activated at -55 +/- 5 mV and -24 +/- 5 mV respectively, corresponding to low (LVA) and high (HVA) voltage-activated currents. The proportion of cells displaying such current separation was reduced either at voltage gradients of 0.053 V/s or 1.6 V/s. The application of nitrendipine and nicardipine resulted in a blockage of both currents, where the HVA current component was more sensitive to the agents. The sensitivity of both Ca currents to dihydropyridine antagonists suggests that these currents may be mediated by subtypes of a dihydropyridine sensitive Ca channel.


Innate myeloid cell TNFR1 mediates first line defence against primary Mycobacterium tuberculosis infection.
TNF is crucial for controlling Mycobacterium tuberculosis infection and understanding how will help immunomodulating the host response. Here we assessed the contribution of TNFR1 pathway from innate myeloid versus T cells. We first established the prominent role of TNFR1 in haematopoietic cells for controlling M. tuberculosis in TNFR1 KO chimera mice. Further, absence of TNFR1 specifically on myeloid cells (M-TNFR1 KO) recapitulated the uncontrolled M. tuberculosis infection seen in fully TNFR1 deficient mice, with increased bacterial burden, exacerbated lung inflammation, and rapid death. Pulmonary IL-12p40 over-expression was attributed to a prominent CD11b(+) Gr1(high) cell population in infected M-TNFR1 KO mice. By contrast, absence of TNFR1 on T-cells did not compromise the control of M. tuberculosis infection over 6-months. Thus, the protective TNF/TNFR1 pathway essential for controlling primary M. tuberculosis infection depends on innate macrophage and neutrophil myeloid cells, while TNFR1 pathway in T cells is dispensable.


Predicting delayed instabilities in viscoelastic solids.
Determining the stability of a viscoelastic structure is a difficult task. Seemingly stable conformations of viscoelastic structures may gradually creep until their stability is lost, while a discernible creeping in viscoelastic solids does not necessarily lead to instability. In lieu of theoretical predictive tools for viscoelastic instabilities, we are presently limited to numerical simulation to predict future stability. In this work, we describe viscoelastic solids through a temporally evolving instantaneous reference metric with respect to which elastic strains are measured. We show that for incompressible viscoelastic solids, this transparent and intuitive description allows to reduce the question of future stability to static calculations. We demonstrate the predictive power of the approach by elucidating the subtle mechanism of delayed instability in thin elastomeric shells, showing quantitative agreement with experiments.


Algae mediate submerged macrophyte response to nutrient and dissolved inorganic carbon loading: a mesocosm study on different species.
Nutrient and dissolved inorganic carbon are two important factors that influence the development and species composition of submerged macrophyte communities in shallow lake ecosystems. Yet little is known about their potential interactive effects on the submerged macrophytes and competition outcome of macrophyte-phytoplankton. We performed a mesocosm experiment to investigate the growth and photosynthetic performance of three submerged macrophytes in relation to phytoplankton/epiphyton with nutrient and bicarbonate enrichment. During the experimental period (42 d), increase in nutrient loading in water column resulted in a substantial burst of phytoplankton and epiphyton growth and increased light attenuation. When combined with nutrient loading, bicarbonate enrichment also resulted in a heavily phytoplankton- and epiphyton-dominated state, although bicarbonate enrichment per se does not cause the phytoplankton and epiphyton growth. However, increase in nutrient loading in water column had significant negative impact on individual performances (growth and photosynthesis) of the three submerged macrophytes and bicarbonate enrichment increased the effect of eutrophication on two dissected-leaf macrophytes (M. spicatum and E. nuttallii). Furthermore, our results also suggest that species-specific photosynthetic performances occurred when submerged macrophytes in an environment with high abundance of phytoplankton/epiphyton. This study highlighted that increase in nutrient loading and bicarbonate in water column is likely to interactively impact both abiotic and biotic properties of a freshwater ecosystem. The interactions of these two factors could select macrophyte species that are able to resist the shading from phytoplankton/epiphyton, making these species more dominant in natural freshwater ecosystems.


Common evolutionary origin of the immune and neuroendocrine systems: from morphological and functional evidence to in silico approaches.
Ten years ago, we surmised that there was a common evolutionary origin for the immune and neuroendocrine systems. This was based on morphological and functional results indicating that a common pool of molecules is shared by the two systems, both in invertebrates and vertebrates. In the past decade, numerous molecular biology experiments have confirmed sequence similarity between invertebrate and vertebrate neuroimmune mediators, such as corticotrophin-releasing hormone. However, sequence similarity-based approaches were inadequate for analyzing other immune-related molecules, such as helical cytokines. This review covers older, and more recent findings on invertebrate immune- and neuroendocrine-related molecules with an evolutionary perspective, and suggests that protein-folding recognition algorithms are a fundamental tool in understanding the evolution of immune- and neuroendocrine-related molecules.


Acute appendicitis caused by metallic foreign body (bullet). Case report.
In a patient who had sustained bullet wounding 9 years previously, acute appendicitis was associated with presence of a bullet in the appendix. The case is described.


[Electric counter shock therapy as a rehabilitation stage for patients with surgically treated acquired heart diseases].
A total of 910 case-reports on electropulse treatment of fibrillation arrhythmia are analyzed in the light of rehabilitating patients operated upon for acquired heart diseases. Experience has shown that re-establishment and retention of the sinus rhythm is of decisive importance in the restitution of physical activity and capacity for work. However, investigations have confirmed that the electropulse therapy helps achieve but good immediate results, whereas the remote ones remain unsatisfactory. The problem of stabilizing the rhythm still awaits its solution. The application of agents blocking the beta-adrenoreceptors has but insignificantly reduced the incidence of recurrent episodes of auricular fibrillation.


Sprouting of striatal serotonin nerve terminals following selective lesions of nigro-striatal dopamine neurons in neonatal rat.
The effects of neonatal intracisternal 6-hydroxydopamine (6-OHDA; 50 micrograms) treatment on striatal serotonin (5-HT) nerve terminals in rat have been characterized using histo- and neurochemical methods. The 6-OHDA lesion caused a 60% reduction of striatal dopamine (DA) concentration when analyzed in the adult stage, while 5-HT levels were increased by about 40% and 3H-5-HT uptake in vitro was increased by about 60%. Using computerized image analysis, a marked increase in 5-HT-like immunoreactive terminal density was found in both rostral (+200%) and caudal (+50%) striatum. Pretreatment with the DA uptake blocker amfolenic acid completely counteracted the 6-OHDA-induced alterations in both DA and 5-HT neurons in the striatum, while pretreatment with the noradrenaline uptake blocker desipramine had no significant effects. Regional analysis of 5-HT levels in the CNS after neonatal 6-OHDA treatment or the combined desipramine + 6-OHDA treatment showed no significant effect in any of the brain areas analyzed, apart from the observed 5-HT increase in striatum. It was furthermore observed that the striatal 5-hydroxyindoleacetic acid (5-HIAA)/5-HT ratio was decreased, while the 3,4-dihydroxyphenylacetic acid (DOPAC)/DA ratio was increased following the 6-OHDA lesion, indicating compensatory mechanisms in turnover of transmitters. These alterations were completely reversed after pretreatment with amfolenic acid. The present results support the view that the 5-HT hyperinnervation following neonatal 6-OHDA treatment is a collateral sprouting response induced by lesioning of the striatal DA neurons.


Advances in assays of matrix metalloproteinases (MMPs) and their inhibitors.
Matrix metalloproteinases (MMPs) play an important role in many physiological and pathological processes. To assay the activities of MMPs is important in diagnosis and therapy of the MMPs associated diseases, such as neoplastic, rheumatic and cardiovascular diseases. Several assay systems have been developed, which include bioassay, zymography assay, immunoassay, fluorimetric assay, radio isotopic assay, phage-displayed assay, multiple-enzyme/multiple-reagent assay and activity-based profiling assay. The principle, application, advantage and disadvantage of these assays have been reviewed in this article.


Patient-reported Outcomes from JAVELIN Bladder 100: Avelumab First-line Maintenance Plus Best Supportive Care Versus Best Supportive Care Alone for Advanced Urothelial Carcinoma.
In JAVELIN Bladder 100, avelumab first-line maintenance plus best supportive care (BSC) significantly prolonged overall survival (OS; primary endpoint) versus BSC alone in patients with advanced urothelial carcinoma (aUC) without disease progression with first-line platinum-containing chemotherapy. To evaluate patient-reported outcomes (PROs) with avelumab plus BSC versus BSC alone. A randomized phase 3 trial (NCT02603432) was conducted in 700 patients with locally advanced or metastatic urothelial carcinoma that had not progressed with first-line gemcitabine plus cisplatin or carboplatin. PROs were a secondary endpoint. Avelumab plus BSC (n = 350) or BSC alone (n = 350). National Comprehensive Cancer Network/Functional Assessment of Cancer Therapy Bladder Symptom Index-18 (FBlSI-18) and EuroQol five-level EQ-5D (EQ-5D-5L) assessments were analyzed using descriptive statistics and mixed-effect models. Time to deterioration (TTD; prespecified definition: a ≥3-point decrease from baseline in the FBlSI-18 disease-related symptoms-physical subscale for two consecutive assessments) was evaluated via Kaplan-Meier analyses. Completion rates for scheduled on-treatment PRO assessments were >90% (overall and average per assessment). Results from descriptive analyses and mixed-effect or repeated-measures models of FBlSI-18 and EQ-5D-5L were similar between arms. TTD was also similar, both in the prespecified analysis (hazard ratio 1.26 [95% confidence interval: 0.90, 1.77]) and in the post hoc analyses including off-treatment assessments and different event definitions. Limitations included the open-label design and limited numbers of evaluable patients at later time points. Addition of avelumab first-line maintenance to BSC in patients with aUC that had not progressed with first-line platinum-containing chemotherapy prolonged OS, with a relatively minimal effect on quality of life. In this trial of people with advanced urothelial carcinoma who had benefited from first-line chemotherapy (ie, had stable disease or reduced tumor size), treatment with avelumab maintenance plus best supportive care (BSC) versus BSC alone improved survival significantly, without compromising quality of life, as reported by the patients themselves.


Factors associated with return-to-work and health outcomes among survivors of road crashes in Victoria.
To explore the relationships between injury, disability, work role and return-to-work outcomes following admission to hospital as a consequence of injury sustained in a road crash. Prospective cohort study of patients admitted to an adult trauma centre and two metropolitan teaching hospitals in Victoria, Australia. Participants were interviewed in hospital, 2.5 and eight months post-discharge. Participants were 60 employed and healthy adults aged 18 to 59 years admitted to hospital in the period February 2004 to March 2005. Despite differences in health between the lower extremity fracture and non-fracture groups eight months post-crash the proportions having returned to work was approximately 90%. Of those returning to work, 44% did so in a different role. After adjustment for baseline parameters, lower extremity injuries were associated with a slower rate of return to work (HR: 0.31; 95%CI: 0.16-0.58) as was holding a manual occupation (HR: 0.16; 95%CI: 0.09-0.57). There were marked differences in physical health between and within the injury groups at both follow-up periods. These results demonstrate that both injury type and severity and the nature of ones occupation have a considerable influence on the rate and pattern of return to work following injury. Further, persisting disability has a direct influence on the likelihood of returning to work. The implications of these findings and the types of data required to measure outcome post-injury are discussed.


Feasability of neat carbon dioxide packed column comprehensive two dimensional supercritical fluid chromatography.
The design and implementation of comprehensive two dimensional supercritical fluid chromatography (SFC) using neat carbon dioxide as the mobile phase is described. Two conventional supercritical fluid chromatographs were hyphenated via an on line comprehensive 2D liquid chromatography like interface; it consisted of a two loop switching valve allowing the collection of the first dimension column effluent, the second dimension separation of a fraction being performed during the time allowed for the collection of the subsequent fraction of the first dimension eluent. Both dimension separations were monitored via UV detection; for the second dimension, the main flow was diverted to implement flame ionisation detection for the detection of hydrocarbons and the construction of the corresponding colour plots. Some key parameters related to the interfacing of the two dimensions and the chromatographic conditions used in both dimensions are discussed. In this preliminary report, the feasibility of comprehensive 2D SFC is demonstrated on synthetic mixtures of hydrocarbons and its potential on real sample analysis is illustrated by the separation of coal derived vacuum distillate.


The Development of an Angiogenic Protein "Signature" in Ovarian Cancer Ascites as a Tool for Biologic and Prognostic Profiling.
Advanced ovarian cancer (AOC) is one of the leading lethal gynecological cancers in developed countries. Based on the important role of angiogenesis in ovarian cancer oncogenesis and expansion, we hypothesized that the development of an "angiogenic signature" might be helpful in prediction of prognosis and efficacy of anti-angiogenic therapies in this disease. Sixty-nine samples of ascitic fluid- 35 from platinum sensitive and 34 from platinum resistant patients managed with cytoreductive surgery and 1st-line carboplatin-based chemotherapy- were analyzed using the Proteome ProfilerTM Human Angiogenesis Array Kit, screening for the presence of 55 soluble angiogenesis-related factors. A protein profile based on the expression of a subset of 25 factors could accurately separate resistant from sensitive patients with a success rate of approximately 90%. The protein profile corresponding to the "sensitive" subset was associated with significantly longer PFS (8 [95% Confidence Interval {CI}: 8-9] vs. 20 months [95% CI: 15-28]; Hazard ratio {HR}: 8.3, p<0.001) and OS (20.5 months [95% CI: 13.5-30] vs. 74 months [95% CI: 36-not reached]; HR: 5.6 [95% CI: 2.8-11.2]; p<0.001). This prognostic performance was superior to that of stage, histology and residual disease after cytoreductive surgery and the levels of vascular endothelial growth factor (VEGF) in ascites. In conclusion, we developed an "angiogenic signature" for patients with AOC, which can be used, after appropriate validation, as a prognostic marker and a tool for selection for anti-angiogenic therapies.


Comparative evaluation of indirect and sandwich ELISA for the detection of antibodies to bovine respiratory syncytial virus (BRSV) in dairy cattle.
Seroprevalence of bovine respiratory syncytial virus (BRSV) infection in both exotic and crossbred cattle were described. A baculovirus expressed recombinant purified nucleocapsid (N) protein was used in indirect and sandwich ELISA for screening of 499 bovine sera samples from all over the state for the presence of BRSV antibodies. The seroprevalence rate of BRSV was found to be 46.09% through indirect ELISA while it would found to be 65.33% by sandwich ELISA. The result also indicated that exotic breeds were more susceptible to BRSV infection compared to crossbred cattle. A comprehensive analysis on susceptibility to BRSV as regards to various factors like age and sex was also summarized.


Ultrasensitive visual detection of miRNA-143 using a CRISPR/Cas12a-based platform coupled with hyperbranched rolling circle amplification.
MicroRNAs are proposed novel biomarker for noninvasive diagnosis of cancer. miRNA-143 is reported to be associated with the development of prostate cancer. However, detection of miRNAs is still challenging due to their unique characteristics, such as small size and high sequence homology among family members. We here developed a gold nanoparticle (AuNP)-based visual assay that combines with CRISPR/Cas12a-assisted hyperbranched rolling circle amplification (HRCA), which is called HRCA enhanced CRISPR/Cas12a-based assay (HECA) for sensitive detection of miRNA-143. The sequence-specific recognition character of CRISPR/Cas12a and HRCA signal amplification strategy enables the HECA outstanding specificity and sensitivity. In optimal condition, 1 fM miRNA-143 could be detected by naked eyes, and down to aM level with the aid of UV-Vis instrument. The diagnostic performance of the HECA for clinical samples was also evaluated based on the receiving operating characteristic algorithm (ROC), and our results suggest the miR-143 is a promising biomarker for noninvasive diagnosis of prostate cancer. This method is simple in operation and requires minimum instrument. We expect it to be widely applied in clinical diagnostics, especially in low-resource settings.


Do non-contact injuries occur during high-speed running in elite football? Preliminary results from a novel GPS and video-based method.
Understanding how injuries occur (inciting circumstances) is useful for developing etiological hypotheses and prevention strategies. The aims of this study were 1) to evaluate the feasibility of a method combining video and Global Positioning System data to estimate the speed and acceleration of activities leading to injuries and 2) to use this method to analyse the inciting circumstances leading to non-contact injuries. Retrospective descriptive study. Injury inciting circumstances from 46 elite players over three seasons were analysed from video recordings and from external load measures collected through Catapult Vector S7 Global Positioning System. In total 34 non-contact injuries were analysed. Sixteen out of the seventeen hamstring injuries occurred when players were running for (median and interquartile range) 16.75 m (8.42-26.65 m) and achieved a peak speed of 29.28 km·h-1 (26.61-31.13 km·h-1) which corresponded to 87.55 % of players' maximal speed (78.5 %-89.75 %). Of the three adductor injuries, one occurred whilst the player was decelerating without the ball, one occurred whilst the player was accelerating and controlling the ball at knee level, and one occurred whilst the player was performing an instep kick. Two quadriceps injuries occurred whilst the players were kicking either whilst walking or running. From the preliminary results reported in this study most hamstring injuries occurred when players ran >25 km·h-1 and above 80 % of their maximal speed. This study suggests that this novel approach can allow a detailed and standardised analysis of injury inciting circumstances.


CD44 variant 6 is involved in the attachment and invasion of endometrial cells to peritoneum.
To study the effects of CD44 standard (CD44s), CD44v3, and CD44v6 overexpression (OE) on immortalized human endometrial epithelial (iEECs) and stroma cells (human endometrial stromal cells [hESCs]) using in vitro assays and a nude mouse xenograft model. Menstrual endometrial cells from women with endometriosis have increased adhesion and also express higher levels of CD44 variant 6 (v6), but not v3, compared to menstrual endometrial cells from women without endometriosis. In vitro studies and in vivo xenograft model. Academic center. Deidentified immortalized endometrial epithelial tissue samples of a reproductive-age woman. Overexpression of CD44s, CD44v3, and CD44v6 was carried out using lipofectamine, and their expression was verified with mRNA and protein in iEEC and hESCs. The OE cells were used in in vitro studies and an in vivo xenograft model compared to plasmid control. The effect of CD44s, CD44v3, and CD44v6 OE on attachment and invasion assays and a xenograft model with immortalized human stromal and epithelial cells. Expression of mRNA and protein confirmed appropriate OE of CD44s, CD44v3, and CD44v6 in the different cell types. CD44v6 OE increased attachment of hESCs compared with controls. CD44v6 OE did not change the attachment of iEECs. There was no difference in attachment in iEECs or hESCs with OE of CD44s or CD44v3. Overexpression of CD44v6 increases attachment of hESCs to peritoneal mesothelial cells in an in vitro assay and an in vivo xenograft model. Menstrual endometrial cell type and CD44 variants play a complex role in the development of the early endometriotic lesion.


[Fast and slow genome rearrangements: molecular and genetic mechanisms of changes in virulence].
Different mechanisms of virulence changes at the molecular and populational levels are discussed. The methodology and mechanisms of Streptococcus pyogenes variability are analysed. The views of the adaptive role played by insertion sequences are developed, and the design of an epidemiologico-genetic experiment is proposed, which consists in obtaining highly virulent insertion mutants and testing their properties.


A review of the risks and benefits of differing prophylaxis regimens for the treatment of deep venous thrombosis and pulmonary embolism in neurosurgery.
Annually, 2 million people in the United States develop deep venous thrombosis (DVT), and nearly 100,000 sustain fatal pulmonary emboli. Prophylaxis against DVT/pulmonary embolism (PE) is a critical issue, and options include elastic stockings, intermittent pneumatic compression stockings, low-dose unfractionated heparin (5000 U every 8-12 hours), and low molecular-weight heparin (ie, enoxaparin and dalteparin). The risks and benefits associated with different prophylaxis regimens used in the prevention of DVT and PE in neurosurgical procedures were analyzed. Neurosurgical studies focusing on different methods of prophylaxis used for the prevention of DVT and PE were reviewed. The efficacy, risks, and benefits of varied treatment options were evaluated, with particular emphasis on minor and major hemorrhages occurring where heparin-based protocols were used. In Flinn et al series (Arch Surg. 1996;131(5):472-80), the incidence of DVT was greater for cranial (7.7%) than spinal procedures (1.5%). Although intermittent pneumatic compression devices provided adequate reduction of DVT/PE in some cranial and combined cranial/spinal series, low-dose subcutaneous unfractionated heparin or low molecular-weight heparins further reduced the incidence, not always of DVT, but of PE (Br J Neurosurg 1995;9(2):159-63; J Intensive Care Med 2003;18(2):59-79). Nevertheless, low-dose heparin-based prophylaxis in cranial and spinal series risks minor and major postoperative hemorrhages: 2% to 4% in a cranial series, 3.4% minor and 3.4% major hemorrhages in a combined cranial/spinal series, and a 0.7% incidence of major/minor hemorrhages in a spinal series (J Neurosurg 2003;99(4):680-4; Neurosurgery 1986;18(4):440-5; Eur Spine J 2004;13(1):1-8; J Intensive Care Med 2003;18(2):59-79). Although mechanical prophylaxis provided effective prophylaxis against DVT/PE in many series, the added efficacy of low-dose heparin regimens has to be weighed against risks of major postoperative hemorrhages and their neurological sequelae.


Retrosigmoid approach assisted by high-resolution computed tomography: a cost-effective technique to identify the transverse and sigmoid sinus transition.
When utilizing the retrosigmoid approach (RA), accurately identifying the transverse and sigmoid sinus transition (TSST) is a key procedure for neurosurgeons, especially in developing countries restricted by the lack of expensive devices, such as the neural navigation system and the three-dimensional volumetric image-rendered system. Before operations, a computed tomography scan is a common and cost-effective method of checking patients who suffer lesions located at the cerebellopontine angle. Therefore, we present a technique using only high-resolution computed tomography to identify the transverse and sigmoid sinus transition. This retrospective study included 35 patients who underwent retrosigmoid approach operations to resect an acoustic neurinoma with the assistance of our technique. In brief, our technique contains 4 steps: (1) All patients' 1-mm, consecutive, high-resolution computed tomographic images that clearly displayed landmarks, such as the inion, lambdoid suture, occipitomastoid suture, and the mastoid emissary foramen, were investigated initially. (2) We selected two particular slices (A and B) among all of these high-resolution computed tomographic images in which scanning planes were parallel with the line drawn from the root of the zygoma to the inion (LZI). Slice A contained both the root of the zygoma and the inion simultaneously, and slice B displayed the mastoid emissary foramen. (3) Four points (α, β, γ, δ) were arranged on slices A and B, and point α was located at the inner surface of the skull, which represents the posterior part of the sulci of the sigmoid sinus. Point β was located at the outer surface of the skull, and the line connecting them was perpendicular to the bone. Similarly, on slice B, we labeled point γ as the point that represents the posterior part of the sulci of the sigmoid sinus at the inner surface and point δ as the point located at the outer surface of the skull, and the line connecting them was also perpendicular to the bone. The distances between point β and the lambdoid suture/occipitomastoid suture and between point δ and the mastoid emissary foramen were calculated for slices A and B, respectively. (4) During the operation, a line indicating the LZI was drawn on the bone with ink when the superficial soft tissue was pushed away, and this line would cross the lambdoid suture/occipitomastoid suture. With both the crosspoint and the distance obtained from the high-resolution CT images, we could locate point β. We also used the same method to locate point δ after revealing the mastoid emissary foramen. The line connecting point β and point δ indicated the posterior border of the sigmoid sinus, and the intersection between the line and LZI indicated the inferior knee of the transverse and sigmoid sinus transition (TSST). All 35 patients underwent the RA craniectomies that were safely assisted by our technique, and neither the sigmoid sinus nor the transverse sinus was lacerated during the operations. Our cost-effective technique is reliable and convenient for identifying the transverse and sigmoid sinus transition (TSST) which could be widely performed to guarantee the safety of RA craniectomy.


Effects of treatment of malignant lymphomas on the granulocytic progenitor cells (CFUc).
The goal of this work was to appraise the after-effects of radiotherapy and associated radio- and chemotherapy on hematosarcomas by studying the granulocytic progenitor cells (CFUc). The bone marrow of 14 subjects was cultured in methylcellulose 1--6.5 years after completion of radiotherapy. The results were recorded as the number of CFUc per 10(5) nucleated medullary cells plated and per 10(5) metamyelocytes plated, and the total number of medullary CFUc was calculated. These different modes of expression are discussed. The post-radiotherapeutic effects appeared to be closely correlated with the volum irradiated. Chemotherapy seemed in several cases to accentuate the effects. Nevertheless, the organism can operate a powerful compensatory mechanism by means of increased mitosis, and in addition, certain subjects are able to maintain a normal number of CFUc after an aggressive treatment. It was possible in certain cases to study the medullary regeneration of the irradiated zone by culture of the CFUc and by the 59Fe external counting. Its extent after radiotherapy seemed for both the granulocytic and the red cell line to be correlated with the volume irradiated. Subject to the study of new cases, regeneration appears to be very reduced when chemotherapy is associated with radiotherapy.


Sinonasal adenoid cystic carcinoma: the M. D. Anderson Cancer Center experience.
Adenoid cystic carcinoma of the sinonasal tract is a rare cancer that accounts for 10% of all malignancies at this site. The objective of the current study was to evaluate prognostic factors, treatment outcomes, recurrence patterns, and survival rates for sinonasal adenoid cystic carcinoma. A retrospective chart review was performed at an academic tertiary referral center. Between 1990 and 2004, 105 patients were evaluated for adenoid cystic carcinoma of the sinonasal tract at a single institution. Demographics, presentation, anatomic site, Tumor, Lymph Node, Metastasis (TNM) classification, pathology, treatment, recurrences, and survival were evaluated. One hundred five patients with adenoid cystic carcinoma were evaluated, including 58 women and 47 men. Their median age was 50 years, and the mean follow-up was 47 months. The maxillary sinus (47%) and the nasal cavity (30%) were the most common primary tumor sites. The majority of patients presented with T3/T4 (76.7%), N0 (98%), M0 (97%) disease. Eighty-four percent of patients underwent surgery and received postoperative radiation as treatment for their primary disease. The local recurrence rate was 30%, and the distant metastases rate was 38%. The 5-year overall survival and disease specific survival rates were 62.9% and 70.9%, respectively. Adenoid cystic carcinoma of the paranasal sinuses is a rare disease, and the ideal treatment paradigm has yet to be defined. The current data suggested that surgical resection with postoperative radiation therapy offers durable local control and compares favorably with historic data. Although local recurrences develop in a significant percentage of patients, survival from this disease exceeds that of other sinonasal malignancies.


The E3 ubiquitin ligase NEDD4-1 protects against acetaminophen-induced liver injury by targeting VDAC1 for degradation.
Acetaminophen (APAP) overdose is a major cause of liver injury. Neural precursor cell expressed developmentally downregulated 4-1 (NEDD4-1) is an E3 ubiquitin ligase that has been implicated in the pathogenesis of numerous liver diseases; however, its role in APAP-induced liver injury (AILI) is unclear. Thus, this study aimed to investigate the role of NEDD4-1 in the pathogenesis of AILI. We found that NEDD4-1 was dramatically downregulated in response to APAP treatment in mouse livers and isolated mouse hepatocytes. Hepatocyte-specific NEDD4-1 knockout exacerbated APAP-induced mitochondrial damage and the resultant hepatocyte necrosis and liver injury, while hepatocyte-specific NEDD4-1 overexpression mitigated these pathological events both in vivo and in vitro. Additionally, hepatocyte NEDD4-1 deficiency led to marked accumulation of voltage-dependent anion channel 1 (VDAC1) and increased VDAC1 oligomerization. Furthermore, VDAC1 knockdown alleviated AILI and weakened the exacerbation of AILI caused by hepatocyte NEDD4-1 deficiency. Mechanistically, NEDD4-1 was found to interact with the PPTY motif of VDAC1 through its WW domain and regulate K48-linked ubiquitination and degradation of VDAC1. Our present study indicates that NEDD4-1 is a suppressor of AILI and functions by regulating the degradation of VDAC1.


Scalemic myrionsumamide A, tetracyclic skeleton alkaloids from Myrioneuron effusum.
Investigation of the alkaloids from Myrioneuron effusum leads to the isolation of myrionsumamide A (1), a pair of enantiomeric alkaloids with an unprecedented tetracyclic system skeleton. These two alkaloids were separated by chiral HPLC with a ratio of 3 : 5 from the scalemic mixture. Their structures including absolute configurations were determined by NMR spectroscopy, X-ray diffraction data and ECD calculations. Both (+)-1 and (-)-1 showed antibacterial activity against Staphylococcus aureus with MIC at 7.81 μg ml-1.


A human metallothionein pseudogene containing AG/CT repetitive elements.
A lambda phage recombinant clone, 25 S, which contains a 15.5-kb EcoRI human genomic DNA fragment, has been characterized. Restriction mapping and Southern blot hybridization indicated a 3.0-kb HindIII fragment containing metallothionein (MT)-like sequences. Several interesting features were found upon comparison of this nucleotide sequence with that of other human MT genes: (1) sequences representing the 5' regulatory region, the 5' untranslated region, and the first exon are not contained in the 3.0-kb HindIII fragment; (2) the coding sequence of the second exon (amino acids 10-31 encoding a portion of the beta-domain of the MT protein) has 11 amino acid changes out of a total of 21, whereas, the third exon (amino acids 32-61, representing the complete alpha-domain of the MT protein) has only 4 amino acid substitutions; however, all cysteine residues are conserved; (3) this MT-like gene retains intron sequences and processing signals; (4) Southern blot analysis of human genomic DNA indicated this MT-like gene is located on a 10.5-kb EcoRI genomic DNA fragment; and (5) unusual AG/CT-rich repetitive elements are located within the second intron and upstream of the second exon of this MT-like gene. This gene is not expressed in response to metal induction in two human cell lines, as shown by northern blot analyses. Based on these observations, this MT-like gene represents a unique nonprocessed pseudogene of the human MT multigene family.


Structural determinants of the noncatalytic chain of tissue-type plasminogen activator that modulate its association rate with plasminogen activator inhibitor-1.
In order to identify the regions of recombinant (r) tissue plasminogen activator (tPA) that mediate its kinetically relevant interaction with r-plasminogen activator inhibitor-1 (rPAI-1), we have determined the second-order association rate (k1) constants of domain-altered variants of tPA with rPAI-1, at 10 degrees C. With two-chain, wild-type recombinant tPA (tcwt-rtPA), obtained by expression of the human cDNA for tPA in five different cell systems (viz. insect cells, human kidney 293 cells, Chinese hamster ovary cells, human melanoma cells, and mouse C127 cells), the average k1 was 1.45 x 10(7) M-1 s-1 (range, 1.34 10(7) M-1 s-1-1.68 x 10(7) M-1 s-1). Since this value was not significantly different for the different tcwt-rtPA preparations, it appears as though the nature of the glycosylation of tPA plays little role in its initial interaction with PAI-1. The k1 determined for tcwt-rtPA was slightly higher than that of 0.87 x 10(7) M-1 s-1, obtained for a similar inhibition of human urokinase by rPAI-1. The k1 value obtained for single-chain (sc) wt-rtPA was approximately 6-fold lower than that of the two-chain molecules, results consistent with previous conclusions on this matter. The k1 value for tcwt-rtPA was not influenced by the presence of epsilon-aminocaproic acid, suggesting that the lysine-binding site associated with the kringle 2 (K2) region of tPA does not modulate the rate of its initial interaction with rPAI-1. Removal of the K2 domain from tPA, by recombinant DNA technology, results in a protein, F-E-K1-P (tc-r delta K2-tPA), containing only the finger (F), growth factor (E), kringle 1 (K1), and serine protease (P) domains. This variant protein was more rapidly inhibited by rPAI-1 (k1 = 3.00 x 10(7) M-1 s-1) than its wild-type counterparts. Deletion of both the K1 and K2 domains resulted in a variant molecule, F-E-P (tc-r delta K1 delta K2-tPA), that was slightly more rapidly inhibited by rPAI-1 (k1 = 2.01 x 10(7) M-1 s-1).(ABSTRACT TRUNCATED AT 400 WORDS)


Mirror movements or functional tremor masking organic tremor.
Functional tremors can be diagnosed based on clinical and physiologic criteria such as entrainment, suggestibility, distractibility, variable nature with the associated clinical history of psychosomatic co-morbidities. The current case report highlights the underrecognized utility of neurophysiology in the correct diagnosis of tremors, providing useful clinical and neurophysiologic insights into clinical and physiological assessment of tremors. A 62-year-old woman with a past medical history of polio was referred by a movement disorders neurologist for evaluation of tremor with concerns of a likely functional etiology. On first assessment there were findings notable for a possible organic etiology, but upon subsequent evaluation the tremor was noted to be variable and entrainable, suggestive of a functional etiology. Neurophysiological tremor study could identify an underlying organic tremor (likely of multi-factorial etiology). Tremor entrainment with contralateral hand tapping could be mirror movements or functional movements, as the underlying organic tremor was not entrained. The amplitude of mirrored movement was commensurate with the tapping amplitude. Functional tremors may mask an underlying organic tremor. Additionally, motor overflow which may happen especially with large amplitude movements may masquerade as mirror movements, which can be difficult to differentiate from an entrained functional tremor. Objective physiology and refinement of the current clinical and physiologic tremor evaluation techniques may help identify an underlying organic etiology.


Effects of arsenic on seed germination and physiological activities of wheat seedlings.
The effects of arsenic (As) were investigated on seed germination, root and shoot length and their biomass and some other factors to elucidate the toxicity of As. The results showed low concentrations of As (0-1 mg/kg) stimulated seed germination and the growth of root and shoot, however, these factors all decreased gradually at high concentrations of As (5-20 mg/kg). The contents of O2*-, MDA, soluble protein and peroxidase (POD) activity all increased with increasing As concentrations. Soluble sugar content, ascorbate peroxidase (APX), and superoxide dismutase (SOD) activities decreased at low concentrations of As, and increased at high concentrations of As. While acetylsalicylic acid (ASA) and chlorophyll contents, catalase (CAT) activity displayed increasing trend when the concentrations of As was lower than 1 mg/kg, and then decreasing trend. By polyacrylamide gel electrophoresis (PAGE), As induced the expression of POD isozymes of wheat seedlings. As induced the expression of CAT isozymes but inhibited the expression of SOD isozymes of wheat seedlings at concentrations lower than 1 mg/kg. However, As inhibited the expression of CAT isozymes but induced the expression of SOD isozymes at concentrations higher than 5 mg/kg. The results indicated As could exert harmfulness in the early development stage of wheat at inappropriate concentrations.


The rat RT1-A1c MHC molecule is a xenogeneic ligand recognized by the mouse activating Ly-49W and inhibitory Ly-49G receptors.
Mouse Ly-49 receptors are known to recognize xenogeneic ligands from hamster and rat. However, until now, there has been no description of a specific rat xenogeneic ligand for any mouse Ly-49 receptor. In this report, we identify RT1-A1c, a rat classical class I MHC molecule, as a ligand for the Ly-49G(BALB/c) inhibitory receptor and the closely related activating receptor, Ly-49W. Xenogeneic class I recognition of targets from PVG but not DA strain rats was mapped to the classical region of the RT1c haplotype by using Con A blasts from RT1c/RT1av1 intra-MHC recombinant rats as targets for RNK-16 cells expressing either Ly-49W or Ly-49G(BALB/c) receptors. Individual expression of class I molecules from PVG and DA rat strains in YB2/0 target cells demonstrate the xenogeneic recognition to be allele specific, because other class I molecules of the RT1c haplotype, RT1-A2c and RT1-U2c, and a classical class I molecule encoded by the RT1av1 haplotype, RT1-Aa, are not recognized by Ly-49W and -G(BALB/c). Furthermore, specificity for RT1-Ac can be transferred from Ly-49W to Ly-49P, which is normally unable to recognize RT1-Ac, by substitution of three residues shared by Ly-49W and -G(BALB/c) but not Ly-49P. These residues are located in the Ly-49 beta4-beta5 loop, which can determine class I allele specificity in mouse Ly-49 receptor interactions with mouse class I ligands, suggesting that mouse Ly-49 recognition of rat class I molecules follows similar principles of interaction. These findings have implications for xenotransplantation studies and for discerning Ly-49 recognition motifs present in MHC molecules.


Geographical variation in India in the composition and lethal potency of Russell's viper (Vipera russelli) venom.
Venom samples of Russell's viper (Vipera russelli) from three localities in India were analysed for their composition and toxicity. Column chromatographic fractionation on CM-Sephadex C-25 showed the absence of three fractions in the venom samples of southern India compared with the samples from northern and western India. The SDS-PAGE pattern of southern Indian venom samples also showed lack of three protein bands corresponding to molecular weights of 66,000, 39,000 and 9000. Venom samples from northern and western India possessed high acidic phospholipase activity while acidic phospholipase activity was absent in the samples from southern India, which in contrast showed large basic fractions with phospholipase activity. Proteolytic activity was present in all the venom samples; however, this activity, as well as trypsin inhibitor activity, was very low in the southern Indian samples. The ratio of proteolytic activity to inhibitor activity remained constant in most of the venom samples studied. LD50 values for most of the venom samples from northern and western India were twice as high as that of the samples from southern India. High phospholipase activity correlated with high lethal potency in the venom samples studied.


Chlorine decay and disinfection by-products transformation under booster chlorination conditions: A pilot-scale study.
Booster chlorination was usually employed in water distribution systems with a long hydraulic retention time. The free chlorine decay and disinfection by-products (DBPs) transformation under booster chlorination conditions were investigated within a pilot-scale water distribution system (WDS). Compared with the initial chlorination in water plants, the loss of chlorine was relatively slow and could be described with first-order kinetic model. The rate of chlorine decay and the generation of DBPs in WDS were much greater than those in beaker. High flow rate and the hydraulic transients both promoted chlorine decay and DBPs formation, especially for dichloroacetonitrile (DCAN). The formation of trihalomethanes (THMs) and haloacetic acids (HAAs) was higher in the ductile iron pipe than in the steel pipe. After booster chlorination, THMs, HAAs, and DCAN all climbed up and then declined continuously, but the peak times were different during the reaction process. The results showed the generation period of DBPs followed the order: THMs (27 h) > HAAs (22 h) > DCAN (5 h). DCAN was not stable in WDS and could be decomposed for a long hydraulic retention time (HRT). The decrease of dichloroacetic acid (DCAA) and increase of trichloroacetic acid (TCAA) indicated that DCAA may turn into TCAA. Linear relationships between the free chlorine demand (FCD) and the generation of THMs that considered both buck water and the pipe wall, as well as the different hydraulic conditions, were established to predict the formation of DBPs in WDS after booster chlorination.