Evaluation of total choline from in-vivo volume localized proton MR spectroscopy and its response to neoadjuvant chemotherapy in locally advanced breast cancer.
Results of the proton magnetic resonance spectroscopy carried out on normal, benign breast disease and locally advanced breast cancer patients are presented. The in-vivo MR spectra of malignant breast tissue of patients (n = 67) suffering from infiltrating ductal carcinoma are dominated by the water resonance, while the spectra of the unaffected contralateral breast tissue of these patients are mainly dominated by resonance arising from lipids which is similar to the spectra of normal breast tissue obtained from volunteers (controls, n = 16). In addition to the water and lipid peaks, in majority of the patients (approximately 80%) the water suppressed spectra showed a resonance at 3.2 ppm due to choline containing compounds (TCho) before treatment. In patients receiving neoadjuvant chemotherapy, absence/reduction in choline was observed in 89% of the patients. TCho was also observed in 2 of 14 benign lesions. The sensitivity and specificity of in-vivo MRS in detecting TCho in malignant tumours was 78% and 86%, respectively. Observation of TCho before treatment and its disappearance (or reduction) after treatment may be a useful indicator of response of locally advanced breast cancer to neoadjuvant chemotherapy.


Surface modification of coronary stents with SiCOH plasma nanocoatings for improving endothelialization and anticoagulation.
The surface properties of intravascular stent play a crucial role in preventing in-stent restenosis (ISR). In this study, SiCOH plasma nanocoatings were used to modify the surfaces of intravascular stents to improve their endothelialization and anticoagulation properties. SiCOH plasma nanocoatings with thickness of 30-40 nm were deposited by low-temperature plasmas from a gas mixture of trimethysilane (TMS) and oxygen at different TMS:O2 ratios. Water contact angle measurements showed that the SiCOH plasma nanocoating surfaces prepared from TMS:O2 = 1:4 are hydrophilic with contact angle of 29.5 ± 1.9°. The SiCOH plasma nanocoated 316L stainless steel (316L SS) wafers were first characterized by in vitro adhesion tests for blood platelets and human umbilical vein endothelial cells. The in vitro test results showed that the SiCOH plasma nanocoatings prepared from TMS:O2 = 1:4 had excellent hemo- and cytocompatibility. With uncoated 316L SS stents as the control, the SiCOH plasma nanocoated 316L SS stents were implanted into rabbit abdominal artery model for in vivo evaluation of re-endothelialization and ISR inhibition. After implantation for 12 weeks, the animals testing results showed that the SiCOH plasma nanocoatings accelerated re-endothelialization and inhibited ISR with lumen reduction of 26.3 ± 10.1%, which were considerably less than the 41.9 ± 11.6% lumen reduction from the uncoated control group.


Metagenomics of urban sewage identifies an extensively shared antibiotic resistome in China.
Antibiotic-resistant pathogens are challenging treatment of infections worldwide. Urban sewage is potentially a major conduit for dissemination of antibiotic resistance genes into various environmental compartments. However, the diversity and abundance of such genes in wastewater are not well known. Here, seasonal and geographical distributions of antibiotic resistance genes and their host bacterial communities from Chinese urban sewage were characterized, using metagenomic analyses and 16S rRNA gene-based Illumina sequencing, respectively. In total, 381 different resistance genes were detected, and these genes were extensively shared across China, with no geographical clustering. Seasonal variation in abundance of resistance genes was observed, with average concentrations of 3.27 × 1011 and 1.79 × 1012 copies/L in summer and winter, respectively. Bacterial communities did not exhibit geographical clusters, but did show a significant distance-decay relationship (P < 0.01). The core, shared resistome accounted for 57.7% of the total resistance genes, and was significantly associated with the core microbial community (P < 0.01). The core human gut microbiota was also strongly associated with the shared resistome, demonstrating the potential contribution of human gut microbiota to the dissemination of resistance elements via sewage disposal. This study provides a baseline for investigating environmental dissemination of resistance elements and raises the possibility of using the abundance of resistance genes in sewage as a tool for antibiotic stewardship.


Synthesis and evaluation of (S)-4-(3-(2'-[11C]isopropylamino)-2-hydroxypropoxy) -2H-benzimidazol -2-one ((S)-[11C]CGP 12388) and (S)-4-(3-((1'-[18F]-fluoroisopropyl)amino)-2-hydroxypropoxy) -2H- benzimidazol-2-one ((S)-[18F]fluoro-CGP 12388) for visualization of beta-adrenoceptors with positron emission tomography.
The beta-adrenoceptor antagonist (S)-[11C]CGP 12177 (4-(3-(tert-butylamino)-2-hydroxypropoxy)-2H-benzimidazol -2[11C]- one) is a generally accepted radioligand for cardiac and pulmonary PET studies. The synthesis of [11C]CGP 12177 is a laborious and often troublesome procedure. Therefore, (S)-CGP 12388 (4-(3-(isopropylamino)-2-hydroxypropoxy) -2H-benzimidazol-2-one), 5, the isopropyl analogue of CGP 12177, has been labeled with carbon-11 in the isopropyl group via a reductive alkylation by [11C]acetone (3) of the corresponding (S)-desisopropyl compound 2. The fluoro-substituted analogue of (S)-CGP 12388 was prepared by reacting 2 with [18F]fluoroacetone (4). (S)-[11C]CGP 12388 (5) was easily prepared via a one-pot procedure. The radiochemical yield of (S)-[11C]CGP 12388 (600-800 Ci/mmol, EOS) was 18% (EOB) with a total synthesis time of 35 min, whereas (S)-[18F]fluoro-CGP 12388 (6) (> 2000 Ci/mmol, EOS) was synthesized in 105 min with a radiochemical yield of 12% (EOB). Biodistribution studies in rats demonstrated specific binding to beta-adrenoceptors of (S)-[18F]fluoro-CGP 12388 and (S)-[11C]CGP 12388 in lung and heart. The lungs were clearly visualized with PET studies of rats. Total/nonspecific binding at 60 min postinjection was 5.6 for (S)-[11C]CGP 12388 and 2.0 for the (S)-18F compound. Due to its facile synthetic procedure and in vivo data, (S)-[11C]CGP 12388 is a promising beta-adrenoceptor ligand for clinical PET.


Increased expression of glutathione peroxidase 4 strongly protects retina from oxidative damage.
Oxidative damage contributes to cone cell death in retinitis pigmentosa and death of rods, cones, and retinal pigmented epithelial (RPE) cells in age-related macular degeneration. In this study, we explored the strategy of overexpressing components of the endogenous antioxidant defense system to combat oxidative damage in RPE cells and retina. In transfected cultured RPE cells with increased expression of superoxide dismutase1 (SOD1) or SOD2, there was increased constitutive and stress-induced oxidative damage measured by the level of carbonyl adducts on proteins. In contrast, RPE cells with increased expression of glutathione peroxidase 1 (Gpx1) or Gpx4 did not show an increase in constitutive oxidative damage. An increase in Gpx4, and to a lesser extent Gpx1, reduced oxidative stress-induced RPE cell damage. Co-expression of Gpx4 with SOD1 or 2 partially reversed the deleterious effects of the SODs. Transgenic mice with inducible expression of Gpx4 in photoreceptors were generated, and in three models of oxidative damage-induced retinal degeneration, increased expression of Gpx4 provided strong protection of retinal structure and function. These data suggest that gene therapy approaches to augment the activity of Gpx4 in the retina and RPE should be considered in patients with retinitis pigmentosa or age-related macular degeneration.


Immunological control of murine gammaherpesvirus infection is independent of perforin.
Perforin-mediated cytotoxic T cell killing has been suggested to be of importance in the control of noncytopathic virus infections, based on studies with lymphocytic choriomeningitis virus (LCMV). We examined the role of perforin in a mouse model of gammaherpesvirus infection using transgenic perforin-deficient mice. Previous work from this laboratory has shown that CD8 T cells are essential for the resolution of the acute lung infection and control of latently infected B cells in murine gamma-herpesvirus 68 infection. The absence of perforin did not significantly affect the kinetics of either the lytic lung infection or the latent spleen infection. Lymphocytes from both perforin-deficient and control mice secreted comparable levels of IFN-gamma, IL-10 and IL-6. In addition, lymphocytes from both strains had similar levels of CD3epsilon-dependent cytotoxic activity in the spleen, draining lymph nodes and bronchoalveolar lavage. These data indicate that the lack of perforin has little affect on the ability of mice to control an experimental gammaherpesvirus infection.


A pilot study of risk-adapted radiotherapy and chemotherapy in patients with supratentorial PNET.
We undertook this study to estimate the event-free survival (EFS) of patients with newly diagnosed supratentorial primitive neuroectodermal tumor (SPNET) treated with risk-adapted craniospinal irradiation (CSI) with additional radiation to the primary tumor site and subsequent high-dose chemotherapy supported by stem cell rescue. Between 1996 and 2003, 16 patients with SPNET were enrolled. High-risk (HR) disease was differentiated from average-risk (AR) disease by the presence of residual tumor (M(0) and tumor size > 1.5 cm(2)) or disseminated disease in the neuraxis (M(1)-M(3)). Patients received risk-adapted CSI: those with AR disease received 23.4 Gy; those with HR disease, 36-39.6 Gy. The tumor bed received a total of 55.8 Gy. Subsequently, all patients received four cycles of high-dose cyclophosphamide, cisplatin, and vincristine with stem cell support. The median age at diagnosis was 7.9 years; eight patients were female. Seven patients had pineal PNET. Twelve patients are alive at a median follow-up of 5.4 years. The 5-year EFS and overall survival (OS) estimates for all patients were 68% +/- 14% and 73% +/- 13%. The 5-year EFS and OS estimates were 75% +/- 17% and 88% +/- 13%, respectively, for the eight patients with AR disease and 60% +/- 19% and 58% +/- 19%, respectively, for the eight with HR disease. No deaths were due to toxicity. High-dose cyclophosphamide-based chemotherapy with stem cell support after risk-adapted CSI results in excellent EFS estimates for patients with newly diagnosed AR SPNET. Further, this chemotherapy allows for a reduction in the dose of CSI used to treat AR SPNET without compromising EFS.


Influence of photoperiod on puberty in the female pony.
Thirteen 6-8 month-old fillies were assigned to 3 treatment groups: Group 1 had a 16-h fixed daily photoperiod (16L:8D, N = 4), Group 2 a daily photoperiod equivalent to ambient daylength (control, N = 5), and Group 3 a 9-h fixed daily photoperiod (9L:15D, N = 4). The light treatments extended from 17 December to 9 August. Hair shedding occurred first in Group 1 followed by Group 2, then Group 3. The proportions of fillies with 1 or more ovulations (puberty) by the end of the project were 2/4, 5/5, 2/4 in Groups 1, 2 and 3, respectively. The hypothesis that a fixed daily photoperiod which corresponded to maximum daylength at the summer solstice in temperate zones (16 h) would hasten puberty was not supported. Instead, the 16-h light treatment interfered with the attainment of puberty, as indicated by the reduced proportion of fillies ovulating, reduced numbers of ovulations, luteal bodies, and oestrous periods/filly, the extended intervals from the beginning of the project to the first ovulatory oestrus and the shorter period of oestrus. Hair coat changes were dissociated from changes in reproductive activity in Group 1. Puberty also appeared to be retarded in Group 3, but the results were less pronounced than those for Group 1.


Use of solid phase microextraction to estimate toxicity: relating fiber concentrations to body residues--part II.
In the companion paper, solid phase microextraction (SPME) fiber concentrations were used as a dose metric to evaluate the toxicity of hydrophobic pesticides, and concentration-response relationships were found for the hydrophobic pesticides tested in the two test species. The present study extends the use of fiber concentrations to organism body residues to specifically address biotransformation and provide the link to toxic response. Test compounds included the organochlorines p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT), p,p'-dichlorodiphenyldichloroethane (p,p'-DDD), and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE); two pyrethroids, permethrin and bifenthrin; and an organophosphate, chlorpyrifos. Toxicity, body residues, and biotransformation of the target compounds were determined for the midge Chironomus dilutus and the amphipod Hyalella azteca. Significant regression relationships were found without regard to chemical, extent of biotransformation, or whether the chemical reached steady state in the organisms. The equilibrium SPME fiber concentrations correlated with the parent compound concentration in the biota; however, the regressions were duration specific. Furthermore, the SPME fiber-based toxicity values yielded species-specific regressions with the parent compound-based toxicity values linking the use of SPME fiber as a dose metric with tissue residues to estimate toxic response.


The Anthrax Vaccine Program: an analysis of the CDC's recommendations for vaccine use.
The anthrax vaccine was never proved to be safe and effective. It is one cause of Gulf War illnesses, and recent vaccinees report symptoms resembling Gulf War illnesses. The vaccine's production has been substandard. Without adequate evaluation, the Food and Drug Administration recently approved (retrospectively) significant changes made to the vaccine's composition since 1990. The vaccine's mandatory use for inhalation anthrax is "off-label." A skewed review of the vaccine literature by the Centers for Disease Control and Prevention (CDC) led to remunerative collaborative research with the army, involving civilian volunteers. Despite acknowledging possible fetal harm, the CDC offered the vaccine to children and pregnant women. New trends could weaken prelicensure efficacy and safety review of medical products intended for biodefense and avoid manufacturer liability for their use.


The United States experience with the acute and chronic treatment of primary pulmonary hypertension.
Patients who clinically have primary or 'unexplained' pulmonary hypertension are found at autopsy or lung biopsy to have a variety of pulmonary vascular changes, including medial hypertrophy, thrombosis, intimal fibrosis and plexiform lesions. It is not surprising that the haemodynamic response to vasodilators varies widely. In general, the non-specific vasodilators used to treat pulmonary hypertension cause an acute fall in systemic arterial pressure, with an increase in cardiac output and a reduction in pulmonary vascular resistance. Pulmonary arterial pressure usually does not change much but occasionally drops dramatically. The risk of death in an acute trial of a vasodilator is less than 0.5% in experienced hands. The use of a short-acting vasodilator (e.g., prostacyclin) may indicate the presence or absence of vasoconstriction, the likelihood of fixed structural obstruction to flow and the risk of administering longer-acting vasodilators, and it may give a clue to prognosis. The risk-benefit ratio in the use of vasodilators in the long-term treatment of primary pulmonary hypertension needs to be evaluated by a controlled trial, conducted in those who respond acutely. The role of high-dose calcium channel blocker treatment and multiple drug therapy will also require further study.


Pediatric influenza.
Influenza viruses cause annual epidemics in the United States. Although the severity of influenza epidemics vary by season, the morbidity associated with annual influenza epidemics in children is considerable from year to year. Excess pediatric outpatient clinic visits, emergency department visits, hospitalizations, and deaths occur each influenza season and are more common among younger children and those with conditions that increase their risk for developing influenza-related complications. Vaccination is the most effective way to prevent influenza and its complications and is recommended for all children 6 months through 18 years of age. Antiviral treatment is another tool to prevent influenza and reduce the duration of illness and complications. This article will review the virological, clinical, and epidemiological features of seasonal influenza, as well as discuss seasonal influenza vaccination and antiviral therapy.


Decreased serum levels of brain-derived neurotrophic factor in adults with attention-deficit hyperactivity disorder.
It has been hypothesized that brain-derived neurotrophic factor (BDNF) is involved in the pathogenesis of attention-deficit hyperactivity disorder (ADHD), although experimental data regarding the contribution of BDNF gene polymorphisms to this psychiatric disorder are controversial. Recently, changes in BDNF serum levels have been reported in children with ADHD, but there are no studies about the possible role of this neurotrophin in adults. A total of 54 Caucasoid ADHD adults, including the predominantly inattentive and combined types (aged 33.43 ± 8.99 yr) and 59 Caucasoid unrelated healthy controls (aged 35.52 ± 9.37 yr) were included in a study to evaluate BDNF levels in serum. Medical, neurological and psychiatric co-morbidities were excluded. Clinical data concerning ADHD diagnosis and blood samples for patients and controls were collected. BDNF serum levels were significantly lower in adults with ADHD compared to healthy controls (p < 0.0001). Although the combined type of ADHD subgroup displayed lower BDNF serum levels than the inattentive type, the differences did not reach statistical significance. No significant correlations were found between serum BDNF levels and scores on the Conners' Adult ADHD Rating Subscales. These results suggest a role for BDNF in ADHD, at least in those patients whose disorder persists throughout life. Low BDNF levels may contribute to the neurodevelopmental deficits of ADHD and to the persistence of the disorder into adulthood. BDNF differences between ADHD subtypes should be further studied.


Facile Synthesis of Colloidal Lead Halide Perovskite Nanoplatelets via Ligand-Assisted Reprecipitation.
In this work, we demonstrate a facile method for colloidal lead halide perovskite nanoplatelet synthesis (Chemical formula: L2[ABX3]n-1BX4, L: butylammonium and octylammonium, A: methylammonium or formamidinium, B: lead, X: bromide and iodide, n: number of [BX6]4- octahedral layers in the direction of nanoplatelet thickness) via ligand-assisted reprecipitation. Individual perovskite precursor solutions are prepared by dissolving each nanoplatelet constituent salt in N,N-dimethylformamide (DMF), which is a polar organic solvent, and then mixing in specific ratios for targeted nanoplatelet thickness and composition. Once the mixed precursor solution is dropped into nonpolar toluene, the abrupt change in the solubility induces the instantaneous crystallization of nanoplatelets with surface-bound alkylammonium halide ligands providing colloidal stability. Photoluminescence and absorption spectra reveal emissive and strongly quantum-confined features. X-ray diffraction and transmission electron microscopy confirm the two-dimensional structure of the nanoplatelets. Furthermore, we demonstrate that the band gap of perovskite nanoplatelets can be continuously tuned in the visible range by varying the stoichiometry of the halide ion(s). Lastly, we demonstrate the flexibility of the ligand-assisted reprecipitation method by introducing multiple species as surface-capping ligands. This methodology represents a simple procedure for preparing dispersions of emissive 2D colloidal semiconductors.


Comparative proteomic analysis of proteins in response to simulated acid rain in Arabidopsis.
A proteomic study using 2-D gel electrophoresis and MALDI-TOF MS was performed to characterize the responses of Arabidopsis thaliana plants to simulated acid rain (SiAR) stress, which is a global environmental problem and has become a serious issue in China in recent years. The emphasis of the present study was to investigate the overall protein expression changes when exposed to SiAR. Out of over 1000 protein spots reproducibly resolved, 50 of them changed their abundance by at least 2-fold. Analysis of protein expression patterns revealed that a set of proteins associated with energy production, metabolism, cell rescue, cell defense and protein folding, etc., could play important roles in mediating plant response to SiAR. In addition to this, some proteins involved in stress responses and jasmonic acid pathway are also involved in plant response to SiAR. More interestingly, the expression of several ubiquitination-related proteins changed dramatically after 32-h SiAR treatment, suggesting that they may act as a molecular marker for the injury phenotype caused by SiAR. Based on our results, we proposed a schematic model to explain the mechanisms associated with the systematic response of Arabidopsis plants to SiAR.


[Infrapopliteal angioplasty: a forgotten region?].
Although percutaneous transluminal angioplasty (PTA) has become established as an endovascular technique--in the more strict sense as a balloon dilatation but being increasingly supplemented by stent implantations--for many vascular regions, reports on infrapopliteal angioplasties are rather scarce. With the development of hydrophilically coated guide wires, improved catheter materials, and dedicated balloon catheters for infrapopliteal use, dilatation treatments distal of the popliteal artery are now standard procedures. Major advances in peri-interventional drug management have also made their contribution. The initial technical and clinical results obtained are excellent. For the majority of the patients, the main concern is for limb salvage. In this context, infrapopliteal PTA also achieves satisfactory results. However, the long-term results must still be considered as unsatisfactory since recurrences are frequent and require repeat interventions. Therefore new therapeutic strategies are required that can reduce re-stenoses especially in this peripheral vascular segment.


[Prospective national study of complicated diverticulitis in Great Britain].
Three hundred cases of complicated diverticular disease were entered into a national audit organised by the Surgical Research Society from 30 hospitals between 1985 and 1988. Admission complications included acute phlegmon (n = 104), pericolic abscess (n = 34), purulent peritonitis (n = 40), large bowel obstruction (n = 31), faecal peritonitis (n = 23), pericolic abscess complicated by fistula (n = 28) and lower GI bleeding (n = 40). The overall mortality was only 11% (4% acute phlegmon, 27% purulent peritonitis, 12% pericolic abscess, 48% faecal peritonitis, 6% large bowel obstruction, 2.5% bleeding, 3.5% fistula). Acute phlegmon was managed non-operatively in 75% and by resection in 23%. Purulent peritonitis was generally managed by Hartmann resection (62%) or by resection and primary anastomosis (15%). Similarly pericolic abscess was usually managed by Hartmann resection (38%) or resection and primary anastomosis (35%). The principal operation for faecal peritonitis was Hartmann resection (83%). Large bowel obstruction was either managed conservatively (13%), by Hartmann resection (29%) or by resection and primary anastomosis with and without a proximal stoma (42%). Most fistulas associated with an abscess were managed by resection and primary anastomosis (82%) and acute GI bleeding was usually managed non-operatively (90%).


Anemia and undernutrition among preschool children in Uttar Pradesh, India.
This study was conducted to assess the prevalence of anemia among preschool children (3-5 years) and its association with malnutrition in rural Barabanki district of Uttar Pradesh, India. Three out of 18 sub-centers in Nindura block, Barabanki, each with six villages, were randomly selected for this survey and 654 boys and 546 girls were included. Mean hemoglobin level in g/dL among boys and girls was 10.1 (SD: 1.66) and 9.9 (SD: 1.67) (P <0.06) respectively. The proportion of anemic children (Hb <11 g/dL) was 70%. Boys were heavier and taller as compared to girls. Among the 67.3% underweight children the mean hemoglobin level was 9.85 (SD: 1.67) as compared to 10.39 (SD: 1.62) in those without malnutrition (P <0.0001). Likewise, stunted children (87.6%) had statistically significantly lower mean hemoglobin levels than those not stunted. The odds ratio of an underweight and stunted child having moderate to severe anemia was 1.66. While more than half caretakers knew about the term "anemia " and associated physical weakness with it, only very few (2.5%) knew that iron intake will improve it. They relied on "doctors" (86.7%) for anemia prevention.


An experimental study of the effect of prior alcohol consumption on a simulated gambling activity.
Previous survey research with both clinical populations and random samples of the general population has established that individuals may experience harmful impacts arising from both their gambling and their consumption of alcohol. Experimental study of the interaction of alcohol consumption on gambling is notable for its absence from the literature. To experimentally study the interaction of alcohol consumption and gambling behaviour. Participants were randomly allocated into two groups--placebo administered and alcohol administered, thus making an independent samples experimental design. Laboratory. Forty young, male, regular EGM players, who also regularly consumed alcohol. The NEO Personality Inventory; The Scale of Gambling Choices (Revised) (SGC); persistence at gambling while losing, as measured by the number of gambling trials played and amount wagered. Subjects either received a prior intake of three alcoholic drinks each containing approximately 10 g of pure alcohol (beer or wine) or an equal volume of an equivalent non-alcoholic beverage. The alcohol group persisted for twice as many gaming trials as the placebo group with significantly more players who had consumed alcohol losing all their original cash stake (50% compared with 15% of the placebo group). The consumption of alcohol appeared to eliminate the strong associations found in placebo group between individual difference measures and persistence. The analogue game was accepted by participants as a valid form of gambling. The result showed that relatively small quantities of alcohol have a significant effect on the psychological processes that underpin self-control over gambling. This finding challenges the conceptual research paradigm of studying co-morbidity or dual-addicted clinical populations as the most appropriate method of understanding how two addictive behaviours interact.


Outcome of patients treated with a single-fraction dose of palliative radiation for cutaneous T-cell lymphoma.
Cutaneous T-cell lymphoma (CTCL) is a radiosensitive tumor. Presently, treatment with radiation is given in multiple fractions. The current literature lacks data that support single-fraction treatment for CTCL. This retrospective review assesses the clinical response in patients treated with a single fraction of radiation. This study reviewed the records of 58 patients with CTCL, primarily mycosis fungoides, treated with a single fraction of palliative radiation therapy (RT) between October 1991 and January 2011. Patient and tumor characteristics were reviewed. Response rates were compared using Fisher's exact test and multiple logistic regressions. Survival rates were determined using the Kaplan-Meier method. Cost-effectiveness analysis was performed to assess the cost of a single vs a multifractionated treatment regimen. Two hundred seventy individual lesions were treated, with the majority (97%) treated with ≥ 700 cGy; mean follow-up was 41.3 months (range, 3-180 months). Response rate by lesion was assessed, with a complete response (CR) in 255 (94.4%) lesions, a partial response in 10 (3.7%) lesions, a partial response converted to a CR after a second treatment in 4 (1.5%) lesions, and no response in 1 (0.4%) lesion. The CR in lower extremity lesions was lower than in other sites (P=.0016). Lesions treated with photons had lower CR than those treated with electrons (P=.017). Patients with lesions exhibiting large cell transformation and tumor morphology had lower CR (P=.04 and P=.035, respectively). Immunophenotype did not impact response rate (P=.23). Overall survival was significantly lower for patients with Sézary syndrome (P=.0003) and erythroderma (P<.0001). The cost of multifractionated radiation was >200% higher than that for single-fraction radiation. A single fraction of 700 cGy-800 cGy provides excellent palliation for CTCL lesions and is cost effective and convenient for the patient.


Meaning of occupation-based groups for low-income urban youths attending after-school care.
The purpose of this phenomenological study was to describe the meaning of occupational therapy groups focusing on occupational engagement, group process, and social-emotional learning for a purposeful sample of low-income urban youths attending after-school care. Interviews and participant observation were used to study how the children made sense of their experience. Qualitative data analysis resulted in two thematic descriptions of the experience. First, the groups were fun because of engagement in novel and challenging leisure occupations within a supportive group context. Participation in creative activities that allowed choice transformed mood--children experienced happiness and wanted more of these experiences. Second, the participants valued being able to talk about feelings and learn strategies for dealing with anger. Findings provide a glimpse into the possibilities of enhancing occupational balance by engaging children in occupations they find to be fun.


PAF1 cooperates with YAP1 in metaplastic ducts to promote pancreatic cancer.
Acinar-to-ductal metaplasia (ADM) is a precursor lesion of pancreatic ductal adenocarcinoma (PDAC); however, the regulators of the ADM-mediated PDAC development and its targeting are poorly understood. RNA polymerase II-associated factor 1 (PAF1) maintains cancer stem cells leading to the aggressiveness of PDAC. In this study, we investigated whether PAF1 is required for the YAP1-mediated PDAC development and whether CA3 and verteporfin, small molecule inhibitors of YAP1/TEAD transcriptional activity, diminish pancreatic cancer (PC) cell growth by targeting the PAF1/YAP1 axis. Here, we demonstrated that PAF1 co-expresses and interacts with YAP1 specifically in metaplastic ducts of mouse cerulein- or KrasG12D-induced ADM and human PDAC but not in the normal pancreas. PAF1 knockdown (KD) reduced SOX9 in PC cells, and the PC cells showed elevated PAF1/YAP1 complex recruitment to the promoter of SOX9. The PAF1 KD reduced the 8xTEAD and SOX9 promoter-luciferase reporter activities in the mouse KC (KrasG12D; Pdx-1 Cre) cells and human PC cells, indicating that the PAF1 is required for the YAP1-mediated development of ADM and PC. Moreover, treatment with CA3 or verteporfin reduced the expressions of PAF1, YAP1, TEAD4, and SOX9 and decreased colony formation and stemness in KC and PC cells. CA3 treatment also reduced the viability and proliferation of PC cells and diminished the duct-like structures in KC acinar explants. CA3 or verteporfin treatment decreased the recruitment of the PAF1/YAP1 complex to the SOX9 promoter in PC cells and reduced the 8xTEAD and SOX9 promoter-luciferase reporter activities in KC and PC cells. Overall, PAF1 cooperates with YAP1 during ADM and PC development, and verteporfin and CA3 inhibit ADM and PC cell growth by targeting the PAF1/YAP1/SOX9 axis in vitro and ex vivo models. This study identified a regulatory axis of PDAC initiation and its targeting, paving the way for developing targeted therapeutic strategies for pancreatic cancer patients.


The natural progression of kidney injury in young type 1 diabetic patients.
Diabetes is the most common cause of end-stage renal disease in industrialized countries. This article describes the structural changes in early diabetic nephropathy and the relationship with renal functional parameters, blood pressure, and albumin excretion. The detrimental influence of sustained hyperglycemia and/or glycemic fluctuations on renal structural change has been well documented. Tight glycemic control is paramount to preventing the development, and even the regression, of renal lesions. As much of the renal injury from diabetes occurs in clinical silence before symptoms or laboratory findings of renal injury are evident, finding early markers of risk is imperative so that nephropathy can be prevented. Currently, the only clinical surrogate marker of diabetic renal injury available is microalbuminuria. However, given the reports of regression of microalbuminuria back to normoalbuminuria, the reliability of this tool as an indicator of risk has been questioned. The need for alternative, noninvasive surrogate markers is described in this report.


Presence of radiolabelled metabolites in release studies using [3H]gamma-aminobutyric acid.
Most studies on gamma-aminobutyric acid (GABA) release from nervous tissue have been conducted using radiolabelled GABA in the presence of aminooxyacetic acid (AOAA) to inhibit GABA: 2-oxoglutarate aminotransferase (GABA-T) to prevent conversion of labelled GABA to labelled catabolites. Here we present data showing that even in the presence of 10 microM-AOAA the spontaneous release of tritium from rat cortical synaptosomes prelabelled with 2,3-[3H]GABA is mainly in the form of tritiated water but that the increase in tritium release in the presence of unlabelled GABA or high potassium-ion concentrations is in the form of authentic [3H]GABA. Interpretation of results should take these facts into account.


Intra-Rater, Inter-Rater and Test-Retest Reliability of an Instrumented Timed Up and Go (iTUG) Test in Patients with Parkinson's Disease.
The "Timed Up and Go" (TUG) is a widely used measure of physical functioning in older people and in neurological populations, including Parkinson's Disease. When using an inertial sensor measurement system (instrumented TUG [iTUG]), the individual components of the iTUG and the trunk kinematics can be measured separately, which may provide relevant additional information. The aim of this study was to determine intra-rater, inter-rater and test-retest reliability of the iTUG in patients with Parkinson's Disease. Twenty eight PD patients, aged 50 years or older, were included. For the iTUG the DynaPort Hybrid (McRoberts, The Hague, The Netherlands) was worn at the lower back. The device measured acceleration and angular velocity in three directions at a rate of 100 samples/s. Patients performed the iTUG five times on two consecutive days. Repeated measurements by the same rater on the same day were used to calculate intra-rater reliability. Repeated measurements by different raters on the same day were used to calculate intra-rater and inter-rater reliability. Repeated measurements by the same rater on different days were used to calculate test-retest reliability. Nineteen ICC values (15%) were ≥ 0.9 which is considered as excellent reliability. Sixty four ICC values (49%) were ≥ 0.70 and < 0.90 which is considered as good reliability. Thirty one ICC values (24%) were ≥ 0.50 and < 0.70, indicating moderate reliability. Sixteen ICC values (12%) were ≥ 0.30 and < 0.50 indicating poor reliability. Two ICT values (2%) were < 0.30 indicating very poor reliability. In conclusion, in patients with Parkinson's disease the intra-rater, inter-rater, and test-retest reliability of the individual components of the instrumented TUG (iTUG) was excellent to good for total duration and for turning durations, and good to low for the sub durations and for the kinematics of the SiSt and StSi. The results of this fully automated analysis of instrumented TUG movements demonstrate that several reliable TUG parameters can be identified that provide a basis for a more precise, quantitative use of the TUG test, in clinical practice.


Molecular cloning and heterologous expression of the isopullulanase gene from Aspergillus niger A.T.C.C. 9642.
Isopullulanase (IPU) from Aspergillus niger A.T.C.C. (American Type Culture Collection) 9642 hydrolyses pullulan to isopanose. IPU is important for the production of isopanose and is used in the structural analysis of oligosaccharides with alpha-1,4 and alpha-1,6 glucosidic linkages. We have isolated the ipuA gene encoding IPU from the filamentous fungi A. niger A.T.C.C. 9642. The ipuA gene encodes an open reading frame of 1695 bp (564 amino acids). IPU contained a signal sequence of 19 amino acids, and the molecular mass of the mature form was calculated to be 59 kDa. IPU has no amino-acid-sequence similarity with the other pullulan-hydrolysing enzymes, which are pullulanase, neopullulanase and glucoamylase. However, IPU showed a high amino-acid-sequence similarity with dextranases from Penicillium minioluteum (61%) and Arthrobacter sp. (56%). When the ipuA gene was expressed in Aspergillus oryzae, the expressed protein (recombinant IPU) had IPU activity and was immunologically reactive with antibodies raised against native IPU. The substrate specificity, thermostability and pH profile of recombinant IPU were identical with those of the native enzyme, but recombinant IPU (90 kDa) was larger than the native enzyme (69-71 kDa). After deglycosylation with peptide-N-glycosidase F, the deglycosylated recombinant IPU had the same molecular mass as deglycosylated native enzyme (59 kDa). This result suggests that the carbohydrate chain of recombinant IPU differed from that of the native enzyme.


Contact-dependent depletion of hydrogen peroxide by catalase is a novel mechanism of myeloid-derived suppressor cell induction operating in human hepatic stellate cells.
Myeloid-derived suppressor cells (MDSC) represent a unique cell population with distinct immunosuppressive properties that have been demonstrated to shape the outcome of malignant diseases. Recently, human hepatic stellate cells (HSC) have been reported to induce monocytic-MDSC from mature CD14(+) monocytes in a contact-dependent manner. We now report a novel and unexpected mechanism by which CD14(+)HLADR(low/-) suppressive cells are induced by catalase-mediated depletion of hydrogen peroxide (H2O2). Incubation of CD14(+) monocytes with catalase led to a significant induction of functional MDSC compared with media alone, and H2O2 levels inversely correlated with MDSC frequency (r = -0.6555, p < 0.05). Catalase was detected in primary HSC and a stromal cell line, and addition of the competitive catalase inhibitor hydroxylamine resulted in a dose-dependent impairment of MDSC induction and concomitant increase of H2O2 levels. The NADPH-oxidase subunit gp91 was significantly increased in catalase-induced MDSC as determined by quantitative PCR outlining the importance of oxidative burst for the induction of MDSC. These findings represent a so far unrecognized link between immunosuppression by MDSC and metabolism. Moreover, this mechanism potentially explains how stromal cells can induce a favorable immunological microenvironment in the context of tissue oxidative stress such as occurs during cancer therapy.


Early Parkinsonism in a Senegalese girl with Lafora disease.
We report the atypical presentation of Lafora disease in a Senegalese girl carrying the homozygous variant, c.560A>C, in the NHLRC1 gene. At 13 years, the patient developed myoclonic and visual seizures, progressive psychomotor slowing, and cognitive decline. At 14 years, a neurological examination showed severe hypomimia, bradykinesia, rigidity and low-amplitude myoclonic jerks. Flash-visual and somatosensory evoked potentials showed an increased amplitude of the cortical components, while an electroretinogram showed attenuated responses. An EEG showed diffuse polyspikes associated with positive-negative jerks as well as posterior slow waves and irregular spikes. The electroclinical picture suggested the diagnosis of Lafora disease regarding the association of visual seizures, cognitive deterioration, and action myoclonus, together with the EEG and evoked potential findings. Two uncommon findings were the prominence of extrapyramidal signs in the early stage of disease (which are rarely reported) and attenuation of electroretinal responses. We consider that Lafora disease should be included in the diagnostic work-up for juvenile Parkinsonism, when associated with epilepsy.


Early childhood mortality from community-acquired infections.
In this study, the authors identified maternal and child characteristics that were independent predictors of death from infectious diseases acquired in the community and determined if these factors could be used to identify groups of children with excess risk of mortality from infection. A historical cohort study was conducted of children less than 5 years of age between 1985 and 1994 (the study period), who were born in Tennessee, and had complete information on their birth certificates. The primary outcome was death from infection identified from death certificates and confirmed through medical record review. Among the 1,014,976 children less than 5 years of age, who contributed 3,351,568 child-years of follow-up, there were 247 deaths from infections (7.4 deaths from infections per 100,000 child-years). Respiratory infections accounted for approximately one half of the deaths. Children having three or more older siblings or birth weight of less than 1,500 g had a 3-fold and 10-fold increased risk of death from infection, respectively, while children with both characteristics had a nearly 20-fold increased risk that persisted beyond the first year of life. Interventions should be focused on prevention of these infections in vulnerable children. At-risk children should be targeted for careful follow-up and early hospitalization when signs of infection develop.


Subtyping of Gliomaby Combining Gene Expression and CNVs Data Based on a Compressive Sensing Approach.
It is realized that a combined analysis of different types of genomic measurements tends to give more reliable classification results. However, how to efficiently combine data with different resolutions is challenging. We propose a novel compressed sensing based approach for the combined analysis of gene expression and copy number variants data for the purpose of subtyping six types of Gliomas. Experimental results show that the proposed combined approach can substantially improve the classification accuracy compared to that of using either of individual data type. The proposed approach can be applicable to many other types of genomic data.


Selected hormonal and neurotransmitter mechanisms regulating feed intake in sheep.
Appetite control is a major issue in normal growth and in suboptimal growth performance settings. A number of hormones, in particular leptin, activate or inhibit orexigenic or anorexigenic neurotransmitters within the arcuate nucleus of the hypothalamus, where feed intake regulation is integrated. Examples of appetite regulatory neurotransmitters are the stimulatory neurotransmitters neuropeptide Y (NPY), agouti-related protein (AgRP), orexin and melanin-concentrating hormone and the inhibitory neurotransmitter, melanocyte-stimulating hormone (MSH). Examination of messenger RNA (using in situ hybridization and real-time PCR) and proteins (using immunohistochemistry) for these neurotransmitters in ruminants has indicated that physiological regulation occurs in response to fasting for several of these critical genes and proteins, especially AgRP and NPY. Moreover, intracerebroventricular injection of each of the four stimulatory neurotransmitters can increase feed intake in sheep and may also regulate either growth hormone, luteinizing hormone, cortisol or other hormones. In contrast, both leptin and MSH are inhibitory to feed intake in ruminants. Interestingly, the natural melanocortin-4 receptor (MC4R) antagonist, AgRP, as well as NPY can prevent the inhibition of feed intake after injection of endotoxin (to model disease suppression of appetite). Thus, knowledge of the mechanisms regulating feed intake in the hypothalamus may lead to mechanisms to increase feed intake in normal growing animals and prevent the wasting effects of severe disease in animals.


Epstein-Barr Virus Positive Diffuse Large B-Cell Lymphoma with Epidermotropism.
Primary cutaneous B-cell lymphomas are rare diseases comprising nodular to diffuse lymphoid cell infiltration with an overlying grenz zone and no epidermal involvement upon histological analysis. Diagnostics can become challenging when lymphomas exhibit the characteristics of both B and T-cells. Differential diagnoses may include reactive proliferations, cutaneous composite lymphomas, and transformed mycosis fungoides. Immunohistochemistry and gene arrangement tests may be beneficial to clarify the diagnosis. Herein, we report a rare case of epidermotropic EBV-positive cutaneous B-cell lymphoma along with a literature review.


Alcohol-Related Liver Disease: Areas of Consensus, Unmet Needs and Opportunities for Further Study.
A joint meeting of the European Association for the Study of the Liver (EASL) and the American Association for the Study of Liver Diseases (AASLD) was held in London on September 30 and October 1, 2017. The goals of the meeting were to identify areas of broad agreement and disagreement, develop consensus, and determine future directions to ultimately reduce the burden, morbidity, and mortality of alcohol-related liver disease (previously termed alcoholic liver disease). The specific aims of the meeting were to identify unmet needs and areas for future investigation, in order to reduce alcohol consumption, develop markers for diagnosis and prognosis of disease, and create a framework to test novel pharmacological agents with pre-specified treatment endpoints. A table summary of these goals and aims is provided in the context of epidemiology, current management strategies, next steps for future trials and translational science.


Thermodynamics of Precision in Markovian Open Quantum Dynamics.
The thermodynamic and kinetic uncertainty relations indicate trade-offs between the relative fluctuation of observables and thermodynamic quantities such as dissipation and dynamical activity. Although these relations have been well studied for classical systems, they remain largely unexplored in the quantum regime. In this Letter, we investigate such trade-off relations for Markovian open quantum systems whose underlying dynamics are quantum jumps, such as thermal processes and quantum measurement processes. Specifically, we derive finite-time lower bounds on the relative fluctuation of both dynamical observables and their first passage times for arbitrary initial states. The bounds imply that the precision of observables is constrained not only by thermodynamic quantities but also by quantum coherence. We find that the product of the relative fluctuation and entropy production or dynamical activity is enhanced by quantum coherence in a generic class of dissipative processes of systems with nondegenerate energy levels. Our findings provide insights into the survival of the classical uncertainty relations in quantum cases.


Plasma levels of soluble tumour necrosis factor receptors are increased in coal miners with pneumoconiosis.
Among other cytokines, tumour necrosis factor (TNF)-alpha is considered to play a key role in the development of mineral dust related fibrosis. Previously, we showed that ex-vivo release of TNF by peripheral blood monocytes is a marker for progression of coal workers' pneumoconiosis (CWP). Since soluble TNF receptors (sTNF-Rs) are believed to play an important regulatory role in systemic effects of TNF, we measured plasma levels of sTNF-R55 and sTNF-R75 in coal miners with (n = 28) or without (n = 76) CWP and in nonexposed controls (n = 29). sTNF-R75 levels were significantly increased in miners with CWP (2.09 +/- 0.44 ng.mL-1) versus the nonexposed controls (1.86 +/- 0.23 ng.mL-1). Neither sTNF-R55 nor sTNF-R75 were related to exposure, stage of pneumoconiosis, smoking, or (spontaneous or ex-vivo induced) monocyte TNF-release. sTNF-R55 was increased in subjects with medication (especially those using cardiovascular drugs); upon exclusion of these subjects, sTNF-R55 was found also to be significantly increased in CWP. In conclusion, bearing in mind a confounding effect of medication, soluble TNF receptors are elevated in plasma of retired miners with coal workers' pneumoconiosis. These observations further support the important role of TNF-mediated pathways in the pathogenesis of mineral dust related fibrosis.


Pickled Vegetables Intake Impacts the Metabolites for Gastric Cancer.
An increased risk of gastric cancer (GC) for pickled vegetables intake has been suggested, but a complete understanding of its pathogenic origin is still lacking, especially from a metabolic viewpoint. We investigated the plasma metabolites and metabolic pathway alteration of GC related to pickled vegetables intake. We analyzed plasma samples collected from 365 gastric cancer patients and 347 healthy individuals, and divided them into three subgroups according to the intake of pickled vegetables. Plasma samples were detected by untargeted metabolomics. Nine metabolites were significantly altered in GC patients among pickled vegetables intake groups (FDR P-value<0.05). All of them were associated with the risk of gastric cancer adjusted for gender, age, smoking status, Helicobacter pylori infection. Pathway analysis showed significant alteration in the folate biosynthesis pathway. In short, we provide new insights from a metabolic perspective on the relationship between pickled vegetables intake and the occurrence of gastric cancer.


The surgical results of the pubovaginal sling procedure using Intravaginal Slingplasty (IVS) for stress urinary incontinence.
We investigated the surgical results and complications of the Intravaginal Slingplasty (IVS) procedure in women with stress urinary incontinence. A total of 72 women with urethral hypermobility underwent the pubovaginal sling procedure using IVS with a mean age of 50.35+/-9.07 years. Average Body Mass Index, mean parity and daily mean pad usage was 29.2+/-3.5, 3.7+/-1.74 and 2.9+/-1.76, respectively. Preoperative urge incontinence was observed in 61.12% of the patients. The mean follow-up was 13.87+/-2.4 months. The operative time, postoperative urge symptoms, de novo detrusor instability and complications were all documented. The mean operating time was 25 min (18-40 min). Though postoperative first monthly controls proved 95.8% total dryness, the cure and improvement rates declined to 45.4% and 24.2% in 66 patients who completed 1 year of follow-up. The overall success and patient satisfaction rates were 69.6% and 87.5% respectively. De novo detrusor instability was observed in five patients (6.9%) whereas detrusor instability persisted in 25% of the patients. Bladder perforation and prolonged urinary retention developed in 11.1% of the patients. No signs of hematoma, infection and erosion were detected. IVS is an easy procedure with low complication rates and it takes about 25 min. In spite of lower cure rates than the alternative surgical techniques, patient satisfaction rates seemed to be similar. Low cure rates may be due to either from inappropriate patient selection or from the biochemical and biomechanical properties of the polypropylene mesh. For this reason, randomized controlled trials and animal experiments should be evaluated for a further decision on the success rates.


The coupling of head, reach and grasp movement in nine months old infant prehension.
In 9-month-old-infants adjustments in the reaching pattern to sudden changes in object location were examined. An attractive ball was presented to the infants at their midline and on some trials (perturbation trials) the ball suddenly changed position 15 cm to the right or left during the reach. For the perturbed trials the movement times approximately doubled compared to the control trials and significantly fewer balls were grasped. The results indicate that infants need to finish the first movement before being able to redirect the reach to a new destination. The correlation between the latency of the head and hand adjustment to the perturbation were 0.85 and 0.78 for movements to the left and to the right, respectively, indicating a tight coupling. The time between the start of the perturbation and peak velocity (TPPV) was significantly shorter for the head movement than for the hand movement, indicating that the head is leading the hand.


Photosynthesis governed by nanoparticulate titanium dioxide. The Pisum sativum L. case study.
Wide availability of anthropogenic TiO2 nanoparticles facilitates their penetration into environment and prompts interactions with plants. They alter plants growth and change their nutritional status. In particular, metabolic processes are affected. In this work the effect of nanometric TiO2 on photosynthesis efficiency in green pea (Pisum sativum L.) was studied. Hydroponic cultivations with three Ti levels (10; 50 and 100 mg L-1) were applied. At all concentrations nanoparticles penetrated into plant tissues and were detected by the single particle ICP-MS/MS method. Nanoparticles altered the CO2 assimilation rate and gas exchange parameters (i.e. transpiration, stomatal conductance, sub-stomatal CO2 concentration). The most pronounced effects were observed for Ti 50 mg L-1 cultivation where photosynthesis efficiency, transpiration and stomatal conductance were increased by 14.69%, 4.58% and 8.92%, respectively. They were further confirmed by high maximum ribulose 1,5-bisphosphate carboxylation rate (27.40% increase), maximum electron transport rate (21.51% increase) and the lowest CO2 compensation point (45.19% decrease). Furthermore, concentrations of Cu, Mn, Zn, Fe, Mg, Ca, K and P were examined with the most pronounced changes observed for elements directly involved in photosynthesis (Cu, Zn, Mn, and Fe). The Cu concentrations in roots, stems and leaves for Ti 50 mg L-1 cultivation were below the control by 33.15%, 38.28% and 10.76%, respectively. The Zn content in analogous treatment and organs decreased by 30.24%, 26.69% and 13.35%. The Mn and Fe levels in leaves were increased by 72.22% and 50.32%, respectively. Our results indicated that plant defence mechanisms which restrain the water uptake have been overcome in pea by photocatalytic activity of nanoparticulate TiO2 which stimulated photosynthesis. On the contrary to the substantial stomatal conductance, the transpiration has been reduced because exceptional part of water flow was already consumed in chloroplasts and could not have been freed to the atmosphere.


Kinetic mechanism of ATP synthesis catalyzed by mitochondrial Fo x F1-ATPase.
Initial rates of succinate-dependent ATP synthesis catalyzed by submitochondrial particles from bovine heart substoichiometrically coupled with oligomycin were found to have hyperbolic dependencies on contents of Mg x ADP, free Mg2+, and phosphate. The results suggest that Mg x ADP complex and free phosphate are true substrates of the enzyme; and an unordered ternary complex of Fo x F1-ATPase, Mg x ADP, and phosphate is generated during the catalysis. The presence of free Mg2+ is required for the reaction. Mg2+ was a noncompetitive activator of ATP synthesis relative to Mg x ADP and a competitive activator relative to phosphate. The decrease in steady-state values of Deltamu(H)+ (by the inhibition of succinate oxidase with malonate) results in the decreased value of Vmax and in a slight decrease in Km for the substrates and Mg2+ without changes in affinity for the substrates. Based on these results, a kinetic scheme of ATP synthesis is proposed.


A Contraction Stress Model of Hypertrophic Cardiomyopathy due to Sarcomere Mutations.
Thick-filament sarcomere mutations are a common cause of hypertrophic cardiomyopathy (HCM), a disorder of heart muscle thickening associated with sudden cardiac death and heart failure, with unclear mechanisms. We engineered four isogenic induced pluripotent stem cell (iPSC) models of β-myosin heavy chain and myosin-binding protein C3 mutations, and studied iPSC-derived cardiomyocytes in cardiac microtissue assays that resemble cardiac architecture and biomechanics. All HCM mutations resulted in hypercontractility with prolonged relaxation kinetics in proportion to mutation pathogenicity, but not changes in calcium handling. RNA sequencing and expression studies of HCM models identified p53 activation, oxidative stress, and cytotoxicity induced by metabolic stress that can be reversed by p53 genetic ablation. Our findings implicate hypercontractility as a direct consequence of thick-filament mutations, irrespective of mutation localization, and the p53 pathway as a molecular marker of contraction stress and candidate therapeutic target for HCM patients.


Enhancement of iron chelation by desferrioxamine entrapped in red blood cell ghosts.
We have exploited the physiologic mechanism for removal of red cells from the circulation to target the iron chelator desferrioxamine to reticuloendothelial iron stores. Compared with free desferrioxamine injected intravenously, the same dose of desferrioxamine entrapped in resealed red blood cell ghosts resulted in a fourfold to fivefold increase in excretion of radioiron in rats with a selective 59Fe radiolabel of reticuloendothelial iron stores. Desferrioxamine in red cell ghosts did not enhance excretion in rats with selective radiolabeling of parenchymal iron stores. In rats with uniformly radiolabeled iron stores, desferrioxamine in red cell ghosts produced an eightfold to ninefold greater loss of iron in the urine free desferrioxamine intravenously or by slow subcutaneous infusion. Desferrioxamine in red cell ghosts resulted in significantly greater fecal excretion of iron than intravenous desferrioxamine, but desferrioxamine in red cell ghosts and subcutaneous desferrioxamine infusion resulted in similar fecal iron excretion. Clinical application of the red cell ghost method for administration of desferrioxamine and other iron chelators may ber useful for improvement of iron chelation efficiency.


A Patient with Berardinelli-Seip Syndrome, Novel AGPAT2 Splicesite Mutation and Concomitant Development of Non-diabetic Polyneuropathy
Primary polyneuropathy in the context of Seip-Berardinelli type 1 seipinopathy, or congenital generalized lipodystrophy type 1 (CGL1) has not been previously reported. We report the case history of a 27 year old female CGL1 patient presenting with an unusual additional development of non-diabetic peripheral neuropathy and learning disabilities in early adolescence. Whole exome sequencing (WES) of the patient genome identified a novel variant, homozygous for a 52 bp intronic deletion in the AGPAT2 locus, coding for 1-acylglycerol-3-phosphate O-acyltransferase 2, which is uniquely associated with CGL1 seipinopathies, with no molecular evidence for dual diagnosis. Functional studies using RNA isolated from patient peripheral blood leucocytes showed abnormal RNA splicing resulting in the loss of 25 amino acids from the patient AGPAT2 protein coding sequence. Stability and transcription levels for the misspliced AGPAT2 mRNA in our patient nonetheless remained normal. Any AGPAT2 protein produced in our patient is therefore likely to be dysfunctional. However, formal linkage of this deletion to the neuropathy observed remains to be shown. The classical clinical presentation of a patient with AGPAT2-associated lipodystrophy shows normal cognition and no development of polyneuropathy. Cognitive disabilities and polyneuropathy are features associated exclusively with clinical CGL type 2 arising from seipin (BSCL2) gene mutations. This case study suggests that in some genetic contexts, AGPAT2 mutations can also produce phenotypes with primary polyneuropathy.


Does attentional training improve numerical processing in developmental dyscalculia?
Recently, a deficit in attention was found in those with pure developmental dyscalculia (DD). Accordingly, the present study aimed to examine the influence of attentional training on attention abilities, basic numerical abilities, and arithmetic in participants who were diagnosed as having DD. Nine university students diagnosed as having DD (IQ and reading abilities in the normal range and no indication of attention-deficit hyperactivity disorder) and nine matched controls participated in attentional training (i.e., video game training). First, training modulated the orienting system; after training, the size of the validity effect (i.e., effect of valid vs. invalid) decreased. This effect was comparable in the two groups. Training modulated abnormalities in the attention systems of those with DD, that is, it reduced their enlarged congruity effect (i.e., faster responding when flanking arrows pointed to the same location as a center arrow). Second, in relation to the enumeration task, training reduced the reaction time of the DD group in the subitizing range but did not change their smaller-than-normal subitizing range. Finally, training improved performance in addition problems in both the DD and control groups. These results imply that attentional training does improve most of the attentional deficits of those with DD. In contrast, training did not improve the abnormalities of the DD group in arithmetic or basic numerical processing. Thus, in contrast to the domain-general hypothesis, the deficits in attention among those with DD and the deficits in numerical processing appear to originate from different sources.


Proteomic Analysis of Zika Virus Infected Primary Human Fetal Neural Progenitors Suggests a Role for Doublecortin in the Pathological Consequences of Infection in the Cortex.
Zika virus (ZIKV) infection is associated with severe neurological defects in fetuses and newborns, such as microcephaly. However, the underlying mechanisms remain to be elucidated. In this study, proteomic analysis on ZIKV-infected primary human fetal neural progenitor cells (NPCs) revealed that virus infection altered levels of cellular proteins involved in NPC proliferation, differentiation and migration. The transcriptional levels of some of the altered targets were also confirmed by qRT-PCR. Among the altered proteins, doublecortin (DCX) plays an important role in NPC differentiation and migration. Results showed that ZIKV infection downregulated DCX, at both mRNA and protein levels, as early as 1 day post infection (1 dpi), and lasted throughout the virus replication cycle (4 days). The downregulation of DCX was also observed in a ZIKV-infected fetal mouse brain model, which displayed decreased body weight, brain size and weight, as well as defective cortex structure. By screening the ten viral proteins of ZIKV, we found that both the expression of NS4A and NS5 were correlated with the downregulation of both mRNA and protein levels of DCX in NPCs. These data suggest that DCX is modulated following infection of the brain by ZIKV. How these observed changes of DCX expression translate in the pathological consequences of ZIKV infection and if other cellular proteins are equally involved remains to be investigated.


Triple negative breast cancer: special histological types and emerging therapeutic methods.
Triple negative breast cancer (TNBC) is a complex and malignant breast cancer subtype that lacks expression of the estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), thereby making therapeutic targeting difficult. TNBC is generally considered to have high malignancy and poor prognosis. However, patients diagnosed with certain rare histomorphologic subtypes of TNBC have better prognosis than those diagnosed with typical triple negative breast cancer. In addition, with the discovery and development of novel treatment targets such as the androgen receptor (AR), PI3K/AKT/mTOR and AMPK signaling pathways, as well as emerging immunotherapies, the therapeutic options for TNBC are increasing. In this paper, we review the literature on various histological types of TNBC and focus on newly developed therapeutic strategies that target and potentially affect molecular pathways or emerging oncogenes, thus providing a basis for future tailored therapies focused on the mutational aspects of TNBC.


Sexual functioning assessed in 4 double-blind placebo- and paroxetine-controlled trials of duloxetine for major depressive disorder.
The onset or worsening of sexual dysfunction is a common treatment-emergent side effect of antidepressant medications. Post hoc analyses of pooled data from placebo-controlled studies were utilized to assess sexual functioning in patients receiving duloxetine or paroxetine. Acute-phase data were obtained from four 8-week, double-blind, placebo- and paroxetine-controlled trials of similar design in which patients meeting DSM-IV criteria for major depressive disorder were randomly assigned to receive placebo (N = 371), duloxetine (40-120 mg/day; N = 736), or paroxetine (20 mg/day; N = 359). Pooling of data from these studies was anticipated during study design. This represented all available data from duloxetine studies in which the Arizona Sexual Experience Scale (ASEX) was administered both at baseline and endpoint. Long-term data were available from extension phases in 2 of these trials in which acute treatment responders received placebo (N = 129), duloxetine (80-120 mg/day; N = 297), or paroxetine (20 mg/day; N = 140) for an additional 26 weeks. Data were collected between March 2000 and July 2002. The incidence of acute treatment-emergent sexual dysfunction was significantly lower among duloxetine-treated patients compared with those receiving paroxetine (p = .015), although both rates were significantly higher than placebo (p = .007 and p < .001 for duloxetine and paroxetine, respectively). Treatment group differences in the incidence of treatment-emergent dysfunction did not vary significantly by gender. In female patients, acute treatment-emergent sexual dysfunction was significantly lower in the duloxetine treatment group compared with the paroxetine treatment group (p = .032), with both rates being significantly higher than placebo (p = .049 and p < .001 for duloxetine and paroxetine, respectively). In the somewhat smaller group of male patients, acute treatment-emergent dysfunction did not differ significantly between duloxetine and placebo treatment groups, but the incidence was significantly higher in paroxetine-treated male patients compared with male placebo patients (p = .012). The long-term incidence of treatment-emergent dysfunction did not differ significantly between duloxetine-, paroxetine-, and placebo-treated patients. In this analysis of pooled data, patients receiving duloxetine (40-120 mg/day) or paroxetine (20 mg/day) had a significantly higher incidence of acute treatment-emergent sexual dysfunction when compared with placebo patients. However, the incidence of acute treatment-emergent dysfunction for duloxetine was significantly lower than that observed for paroxetine.


Treatment of germ cell cancer with two cycles of high-dose ifosfamide, carboplatin, and etoposide with autologous stem-cell support.
To evaluate the activity of two cycles of high-dose ifosfamide, carboplatin, and etoposide (ICE) with autologous hematopoietic progenitor cell support (aHPCS) in patients with poor-prognosis, chemotherapeutically sensitive germ cell cancer. Twenty patients with germ cell tumor who had persistent disease or relapse from standard-risk or high-risk presentation were entered on this pilot study. The entry criteria included relapsed gonadal and extragonadal germ cell cancer unlikely to be cured by standard salvage therapy but without proven refractoriness to chemotherapy. Treatment consisted of two cycles of ICE chemotherapy with mesna uroprotection and aHPCS. On the first cycle, ifosfamide (IFX), 2 gm/m2; carboplatin, 400 mg/m2; and etoposide, 20 mg/kg, were administered on days -6, -5, and -4. On the second cycle, the doses and schedule of carboplatin and etoposide were identical, and patients with normal renal function received additional IFX, 2 g/m2 on day -3 and 1 g/ m2 on day -2. Mesna, 600 mg/m2 every 6 hours, was given until 24 hours following the final dose of IFX on each cycle, and autologous bone marrow and/or peripheral stem-cells were infused on day 0. All twenty patients are assessable for toxicity and current disease status. Two patients received only one cycle of therapy, one because of the development of active hepatitis C following cycle 1, and one because of renal insufficiency. No patient died as a result of protocol therapy, and no patient developed debilitating peripheral neuropathy, symptomatic hearing loss, or severe renal insufficiency requiring dialysis. The median time to recovery of > or = 500 neutrophils/microL and platelets > or = 50,000/microL was day +11 and day +15, respectively. The median maximum creatinine was 1.6 mg/dL on each treatment cycle, and there was no other significant organ toxicity. With a median follow-up of 45 months, nine patients are alive and disease-free following protocol chemotherapy. One patient with embryonal cancer developed progressive pulmonary metastases 3 months after completing high-dose therapy, underwent complete resection of lung metastases, and remains disease-free at 63+ months. Eight patients are continuously disease free at 23+ to 70+ months after protocol therapy. Eleven patients died of progressive disease between 4 and 23 months following completion of treatment. These results compare favorably to other studies in similarly selected patients undergoing salvage therapy with one or two cycles of chemotherapy containing high-dose carboplatin and etoposide with or without cyclophosphamide (CTX) or IFX. The excellent safety and tolerability profile of this regimen and its encouraging activity in poor-prognosis patients make it worthy of further study as part of initial therapy in randomized protocols for high-risk disease and early in the treatment of relapsed germ cell cancer.


Insights into the structural changes occurring upon photoconversion in the orange carotenoid protein from broadband two-dimensional electronic spectroscopy.
Carotenoids play an essential role in photoprotection, interacting with other pigments to safely dissipate excess absorbed energy as heat. In cyanobacteria, the short time scale photoprotective mechanisms involve the photoactive orange carotenoid protein (OCP), which binds a single carbonyl carotenoid. Blue-green light induces the photoswitching of OCP from its ground state form (OCPO) to a metastable photoproduct (OCPR). OCPR can bind to the phycobilisome antenna and induce fluorescence quenching. The photoswitching is accompanied by structural and functional changes at the level of the protein and of the bound carotenoid. Here, we use broadband two-dimensional electronic spectroscopy to study the differences in excited state dynamics of the carotenoid in the two forms of OCP. Our results provide insight into the origin of the pronounced vibrational lineshape and oscillatory dynamics observed in linear absorption and 2D electronic spectroscopy of OCPO and the large inhomogeneous broadening in OCPR, with consequences for the chemical function of the two forms.


Does anterolateral ligament injury change the treatment option in patients with partial ACL tears?
Patients with ACL tears with ALL injury have more clinical complaints (instability, feeling of the pop on the knee or knee sliding). patients have ALL injury with partial ACL tears, It is unclear whether the choice of treatment will be conservative or surgical. This study aimed to determine the effect of anterolateral ligament (ALL) status, whether intact or ruptured, on the choice of conservative or surgical treatment in patients with partial anterior cruciate ligament (ACL) tears. Between 2015 and 2019, patients with suspected partial ACL tears were identified on both physical examination and MR imaging. 122 patients who had partial ACL tears and also status of patient's ALL could be evaluated by radiologist were included in the study, retrospectively. Sixty-two patients who underwent ACL reconstruction were determined as group 1, and 60 patients who did not undergo ACL reconstruction were defined as group 2. In patients with partial ACL rupture with or without ACL reconstruction, it was evaluated whether a ruptured or non- ruptured ALL was effective in this decision of conservative or surgically. The MRIs of patients with partial ACL tears were evaluated by a radiologist and it was concluded that the ALLs of 50 patients were ruptured, and 72 were intact. The ALLs of 36 patients in group 1 were ruptured, and 26 patients were intact. Fourteen patients in group 2 had ruptured ALLs, 46 patient's ALLs were intact. Seventy-two percent of the patients with partial ACL tears who had ruptured ALLs in MRI underwent ACL reconstruction. It was found that ACL reconstruction was performed more frequently in patients with partial ACL tears with ALL rupture. Therefore, we believe that preoperative evaluations of ALLs using MRI in patients with partial ACL tears are essential for surgical planning.


Topical application of a bioadhesive black raspberry gel modulates gene expression and reduces cyclooxygenase 2 protein in human premalignant oral lesions.
Reduced expression of proapoptotic and terminal differentiation genes in conjunction with increased levels of the proinflammatory and angiogenesis-inducing enzymes, cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS), correlate with malignant transformation of oral intraepithelial neoplasia (IEN). Accordingly, this study investigated the effects of a 10% (w/w) freeze-dried black raspberry gel on oral IEN histopathology, gene expression profiles, intraepithelial COX-2 and iNOS proteins, and microvascular densities. Our laboratories have shown that freeze-dried black raspberries possess antioxidant properties and also induce keratinocyte apoptosis and terminal differentiation. Oral IEN tissues were hemisected to provide samples for pretreatment diagnoses and establish baseline biochemical and molecular variables. Treatment of the remaining lesional tissue (0.5 g gel applied four times daily for 6 weeks) began 1 week after the initial biopsy. RNA was isolated from snap-frozen IEN lesions for microarray analyses, followed by quantitative reverse transcription-PCR validation. Additional epithelial gene-specific quantitative reverse transcription-PCR analyses facilitated the assessment of target tissue treatment effects. Surface epithelial COX-2 and iNOS protein levels and microvascular densities were determined by image analysis quantified immunohistochemistry. Topical berry gel application uniformly suppressed genes associated with RNA processing, growth factor recycling, and inhibition of apoptosis. Although the majority of participants showed posttreatment decreases in epithelial iNOS and COX-2 proteins, only COX-2 reductions were statistically significant. These data show that berry gel application modulated oral IEN gene expression profiles, ultimately reducing epithelial COX-2 protein. In a patient subset, berry gel application also reduced vascular densities in the superficial connective tissues and induced genes associated with keratinocyte terminal differentiation.


Longitudinal study of the microbiology of endemic trachoma.
A longitudinal study of trachoma in 100 members of nine Tanzanian families was conducted to assess the sources of variation in the laboratory identification of trachoma and the changes that might occur over time. Multiple conjunctival swabs were collected every 3 months for 1 year and examined by direct fluorescent-antibody cytology (DFA), enzyme immunoassay, or microimmunofluorescence serology for tear antichlamydial antibodies. DFA specimens collected 5 min apart had a discordance rate of 10% and this is attributable to sampling variation. DFA specimens collected 2 days or more apart show a 25% discordance rate. This suggests a biologic variation in shedding in addition to sampling variation. Good correlation existed between the DFA and the enzyme immunoassay. Tear serology was quite specific in predicting the presence of clinical disease and correlated with the other two antigen detection tests, although it does not seem to offer any practical advantages. These studies indicate that there is considerable variation in the shedding of chlamydia by people living in trachoma-endemic areas.


SSTY proteins co-localize with the post-meiotic sex chromatin and interact with regulators of its expression.
In mammals, X- and Y-encoded genes are transcriptionally shut down during male meiosis, but expression of many of them is (re)activated in spermatids after meiosis. Post-meiotic XY gene expression is regulated by active epigenetic marks, which are de novo incorporated in the sex chromatin of spermatids, and by repressive epigenetic marks inherited during meiosis; alterations in this process lead to male infertility. In the mouse, post-meiotic XY gene expression is known to depend on genetic information carried by the male-specific region of the Y chromosome long arm (MSYq). The MSYq gene Sly has been shown to be a key regulator of post-meiotic sex chromosome gene expression and is necessary for the maintenance/recruitment of repressive epigenetic marks on the sex chromatin, but studies suggest that another MSYq gene may also be required. The best candidate to date is Ssty, an MSYq multi-copy gene of unknown function. Here, we show that SSTY proteins are specifically expressed in round and elongating spermatids, and co-localize with post-meiotic sex chromatin. Moreover, SSTY proteins interact with SLY protein and its X-linked homolog SLX/SLXL1, and may be required for localization of SLX/SLY proteins in the spermatid nucleus and sex chromatin. Our data suggest that SSTY is a second MSYq factor involved in the control of XY gene expression during sperm differentiation. As Slx/Slxl1 and Sly genes have been shown to be involved in the XY intra-genomic conflict, which affects the offspring sex ratio, Ssty may constitute another player in this conflict.


The relationship between LDL oxidation and macrophage myeloperoxidase activity.
To explore low density lipoprotein (LDL) oxidation by macrophage myeloperoxidase (MPO) at molecular level. Using a mouse macrophage model, we examined the relationship between LDL oxidation and macrophage MPO by measuring macrophage MPO activity, LDL oxidation products, MPO gene expression and cellular orientation of LDL oxidation. MPO gene expression increased to its maximum gradually when the concentration of LDL was increased, and then maintained at that level. NaN(3) inhibied the elevation of MPO activity and LDL oxidation, which was LDL concentration-dependent. After the composition of macrophage membrane was roughly analyzed, it was determined that the contents of MPO and LDL in 5% sucrose were 7.667 and 21 times higher than those in 10% sucrose, respectively. LDL is attached to the "microdomain" of the macrophage membrane in which LDL is oxidized by MPO.


Optical frequency combs generated by four-wave mixing in optical fibers for astrophysical spectrometer calibration and metrology.
Optical frequency combs generated by multiple four-wave mixing in short and highly nonlinear optical fibers are proposed for use as high precision frequency markers, calibration of astrophysical spectrometers, broadband spectroscopy and metrology. Implementations can involve two optical frequency standards as input lasers, or one standard and a second laser phase-locked to it using a stable microwave reference oscillator. Energy and momentum conservation required by the parametric generation assures phase coherence among comb frequencies, while fibers with short lengths can avoid linewidth broadening and stimulated Brillouin scattering. In contrast to combs from mode-locked lasers or microcavities, the absence of a resonator allows large tuning of the frequency spacing from tens of gigahertz to beyond teraHertz.


Characterization of the genetic diversity, structure, and admixture of 7 Chilean chicken breeds.
The Mapuche fowl is an autochthonous breed raised in Chile and represents an important zoogenetic resource for the local economy. This study aimed at investigating the genetic diversity, relationship and population structure of 96 local Chilean chickens derived from 3 ecotype of Mapuche fowl (Kollonka, Ketro, and Kollonka de aretes), 2 ecotype Chilean (Trintre, Cogote pelado) and 2 breeds (Light Brahma and Barred Plymouth Rock) using 12 microsatellite markers. In total, 113 alleles were detected in all populations, with a mean of 7.6 alleles per population. In all population chicken breeds, the observed and expected heterozygosity ranged from 0.91 to 0.98 and from 0.69 to 0.79. Furthermore, all populations showed significant deviations from Hardy-Weinberg expectations. Across each population, the global heterozygosity deficit (FIT) was -0.174, population differentiation index (FST) was 0.073, and the global inbreeding of individuals within breed (FIS) was -0.267. The phylogenetic relationships of chickens were examined using neighbor-joining trees constructed at the level of population. The highest Nei's standard genetic distance value of 0.559 was observed between Barred Plymouth Rock and Light Brahma, whereas the minimum value (0.099) was found between Kollonka and Trintre. The neighbor-joining tree constructed at population level revealed 2 main clusters, with Light Brahma, Barred Plymouth Rock, Ketro and Kollonka de aretes in 1 cluster, and Kollonka, Trintre and Cogote pelado breeds in the second cluster. Based on the results of the STRUCTURE analysis, the most likely number of clustering of the population evaluated was at K = 3, with Light Brahma and Barred Plymouth Rock breeds forming their own distinct clusters, while Kollonka, Ketro, Kollonka de aretes, Trintre and Cogote pelado breeds clustered together. This study represents the first report of genetic diversity in these populations in Chile. These results can be used as baseline genetic information for genetic conservation program, for instance, to control inbreeding and to implement further genetic studies in local Chilean chickens.


Establishment of a road traffic trauma registry for northern Sri Lanka.
Road traffic injuries are a neglected global public health problem. Over 1.25 million people are killed each year, and middle-income countries, which are motorising rapidly, are the hardest hit. Sri Lanka is dealing with an injury-related healthcare crisis, with a recent 85% increase in road traffic fatality rates. Road traffic crashes now account for 25 000 injuries annually and 10 deaths daily. Development of a trauma registry is the foundation for injury control, care and prevention. Five northern Sri Lankan provinces collaborated with Jaffna Teaching Hospital to develop a local electronic registry. The Centre for Clinical Excellence and Research was established to provide organisational leadership, hardware and software were purchased, and data collectors trained. Initial data collection was modified after implementation challenges were resolved. Between 1 June 2017 and 30 September 2017, 1708 injured patients were entered into the registry. Among these patients, 62% were male, 76% were aged 21-50, 71.3% were motorcyclists and 34% were in a collision with another motorcyclist. There were frequent collisions with uncontrolled livestock (12%) and with fixed objects (14%), and most patients were transported by private vehicles without prehospital care. Head (n=315) and lower extremity (n=497) injuries predominated. Establishment of a trauma registry in low-income and middle-income countries is a significant challenge and requires invested local leadership; the most challenging issue is ongoing funding. However, this pilot registry provides a valuable foundation, identifying unique injury mechanisms, establishing priorities for prevention and patient care, and introducing the concept of an organised system to this region.


The ABCs of MGR with DCJ.
We study the small phylogeny problem in the space of multichromosomal genomes under the double cut and join metric. This is similar to the existing MGR (multiple genome rearrangements) approach but it allows, in addition to inversion and reciprocal translocation, operations of transposition and block interchange. Empirically, with chloroplast and mammalian data sets, it finds solutions as good as or better than MGR when the latter operations are prohibited. Permitting these operations allows quantitatively better solutions where part of the reconstructed ancestral genomes may be included in circular chromosomes. We discuss the biological likelihood of transpositions and block interchanges in the mammalian data.


Internalizing behavior in adolescent girls affects parental emotional overinvolvement: a cross-lagged twin study.
The aim of this study was to examine the direction and the etiology of the association between different parenting styles (parental emotional overinvolvement [EOI] and parental criticism) and internalizing behavior from adolescence to early adulthood. A longitudinal genetically informative cross-lagged design was applied to a population-based sample of Swedish twins contacted at age 16-17 (n = 2369) and at age 19-20 (n = 1705). Sex-limitation modelling revealed different effects for boys and girls. For girls, genetic influences on internalizing problems at age 16-17 independently explained 2.7% of the heritability in parental EOI at age 19-20. These results suggest that emotionally overinvolved and self-sacrificing parental behavior stems in part from daughters (but not sons) genetic predisposition for internalizing behavior. These findings highlight the importance of genetically influenced child-driven effects underlying the parenting-internalizing association, and clarify that the role of such effects may differ depending on sex, type of parenting and developmental period.


Application of genomics and proteomics in drug target discovery.
Gene medicine is making breakthroughs in health questions that have baffled humanity for centuries. To understand and utilize gene medicine, it is necessary to realize its action against targets at the molecular level. Currently, many methods can be used to discover drug targets; among these, genomic and proteomic methods are the two most important. In this study, we introduced how to discover drug targets by genomic and proteomic methods in detail. These contents are beneficial for understanding and utilizing the two methods to discover new drug targets of gene medicine.


The use of altrenogest to control aggression in a male Grant's Zebra (Equus burchelli boehmi).
A male Grant's Zebra (Equus burchelli boehmi) housed with two mares at the Indianapolis Zoo had a 9-yr history of intermittent aggressive behavior toward mares and other animals. Periods of separation allowed the mares time to heal after sustaining superficial bite wounds. On 26 March 2003, the male (890293) was started on altrenogest at a dosage of 19.8 mg orally once daily to allow reintroduction. The dosage was doubled (40 mg once a day) because of a perceived lack of response. Reintroduction to the mares occurred on 17 May 2003 with no signs of aggression noted. Treatment was reduced to 19.8 mg orally once a day and then discontinued. Altrenogest was restarted at 39.5 mg orally once a day because of the planned introduction of a new mare. There have been no major aggressive displays at this dosage of altrenogest and the dosage has recently been reduced following successful introduction of a new mare.


Kurt Semm and the fight against skepticism: endoscopic hemostasis, laparoscopic appendectomy, and Semm's impact on the "laparoscopic revolution".
In the 1970s, Semm developed thermocoagulation, adapted the Roeder Loop, and further invented extra- and intracoporeal endoscopic knotting to achieve endoscopic hemostasis. His numerous technical inventions, especially the electronic insufflator, allowed more complex operations to be performed laparoscopically. His technique, however, was not quickly adopted by the surgical community. When the first fully laparoscopic appendectomy was carried out by Semm in 1980, a veritable storm broke loose. In the opinion of many prominent surgeons, Semm exaggerated the problem of adhesions, and laparoscopic technique itself was regarded as very dangerous. Misunderstood by medical scientists, Semm displayed an ability to force his ideas through despite skepticism and suspicion. He realized that endoscopic surgery had tremendous potential, and promoted laparoscopic technique not only in his field of gynecology but among general surgeons as well. In 1985, Muhe, of Boblingen, Germany, used Semm's technique to remove the first gallbladder in the world laparoscopically. Three years later when Semm presented a videotape of his laparoscopic appendectomy in Baltimore, he gave impetus to McKernan and Save of Marietta. Georgia, to carry out the first laparoscopic cholecystectomy in the United States.


Environmental audits and process flow mapping to assess management of solid waste and wastewater from a healthcare facility: an Italian case study.
In Europe, there are an increasing number of policy and legislative drivers for a more sustainable approach to the management of natural resources as well as for the mitigation of environmental health risks. However, despite significant progress in recent years, there is still some way to go to achieve circularity of process, as well as risk mitigation within organisations. Using a case study of the Gardone Val Trompia hospital in northern Italy, this manuscript offers a novel holistic examination of strategies to enhance resource efficiency and environmental health within a key sector, i.e. the healthcare sector. Through the use of environmental audits and process flow mapping, trends in waste and wastewater arisings and the associated financial and environmental costs and risks were identified. Recommendations for developing more resource efficient approaches as well as mitigating the environmental and public health risks are suggested. These include strategies for improved resource efficiency (including reduction in the hazardous waste) and reduced environmental impacts during the containment, transport and treatment of the waste.


Out-and-back 13C-13C scalar transfers in protein resonance assignment by proton-detected solid-state NMR under ultra-fast MAS.
We present here (1)H-detected triple-resonance H/N/C experiments that incorporate CO-CA and CA-CB out-and-back scalar-transfer blocks optimized for robust resonance assignment in biosolids under ultra-fast magic-angle spinning (MAS). The first experiment, (H)(CO)CA(CO)NH, yields (1)H-detected inter-residue correlations, in which we record the chemical shifts of the CA spins in the first indirect dimension while during the scalar-transfer delays the coherences are present only on the longer-lived CO spins. The second experiment, (H)(CA)CB(CA)NH, correlates the side-chain CB chemical shifts with the NH of the same residue. These high sensitivity experiments are demonstrated on both fully-protonated and 100%-H(N) back-protonated perdeuterated microcrystalline samples of Acinetobacter phage 205 (AP205) capsids at 60 kHz MAS.


Enhanced CD8 T cell responses through GITR-mediated costimulation resolve chronic viral infection.
Chronic infections are characterized by the inability to eliminate the persisting pathogen and often associated with functional impairment of virus-specific T-cell responses. Costimulation through Glucocorticoid-induced TNFR-related protein (GITR) can increase survival and function of effector T cells. Here, we report that constitutive expression of GITR-ligand (GITRL) confers protection against chronic lymphocytic choriomeningitis virus (LCMV) infection, accelerating recovery without increasing pathology. Rapid viral clearance in GITRL transgenic mice coincided with increased numbers of poly-functional, virus-specific effector CD8+ T cells that expressed more T-bet and reduced levels of the rheostat marker PD-1. GITR triggering also boosted the helper function of virus-specific CD4 T cells already early in the infection, as was evidenced by increased IL-2 and IFNγ production, and more expression of CD40L and T-bet. Importantly, CD4-depletion experiments revealed that the expanded pool of virus-specific effector CD8 T cells and the ensuing viral clearance in LCMV-infected GITRL tg mice was entirely dependent on CD4 T cells. We found no major differences for NK cell and regulatory T cell responses, whereas the humoral response to the virus was increased in GITRL tg mice, but only in the late phase of the infection when the virus was almost eradicated. Based on these findings, we conclude that enhanced GITR-triggering mediates its protective, anti-viral effect on the CD8 T cell compartment by boosting CD4 T cell help. As such, increasing costimulation through GITR may be an attractive strategy to increase anti-viral CTL responses without exacerbating pathology, in particular to persistent viruses such as HIV and HCV.


High frequency of the combined presence of QRDR mutations and PMQR determinants in multidrug-resistant Klebsiella pneumoniae and Escherichia coli isolates from nosocomial and community-acquired infections.
Plasmid-mediated quinolone resistance (PMQR) determinants combined with mutations in quinolone resistance-determining regions (QRDRs) and clonal dissemination were investigated in 40 fluoroquinolone-resistant Klebsiella pneumoniae and Escherichia coli isolates from nosocomial and community-acquired infections. We observed nucleotide substitutions in gyrA (Ser83Ile, Val37Leu, Lys154Arg, Ser171Ala, Ser19Asn, Ile198Val, Ser83Tyr, Ser83Leu, Asp87Asn and Asp87Gly) and parC genes (Ser80Ile, Glu84Lys, Ala129Ser, Val141Ala and Glu84Gly). Two novel substitutions were detected in the gyrA gene (Val37Leu and Ile198Val). The presence of PMQR genes predominated in community isolates (55.5 %). In addition to the frequent presence of the class 1 integron in isolates from community-acquired infections, the genetic similarity results obtained by PFGE showed high genomic diversity. This study suggests that management of multidrug-resistant Enterobacteriaceae isolates from the community are a possible source of genetic mobile elements that carry genes that confer resistance to fluoroquinolones. More attention should be paid to the surveillance of community-acquired infections.


Diagnosis and treatment of a fractured third phalanx in a Masai giraffe (Giraffe camelopardalis tippelskirchi).
A 10-yr-old male Masai giraffe (Giraffe camelopardalis tippelskirchi) presented with acute right forelimb lameness. Radiographs revealed a fracture of the medial claw of the distal phalanx penetrating into the distal interphalangeal joint. The giraffe was sedated while it was standing in a chute, and a wooden "hoof block" was applied to the lateral claw of the same limb. The animal was no longer lame 3 days after the procedure. Subsequent treatments included vitamin E, phenylbutazone, and glycosaminoglycans. For 7 wk it was maintained in a small holding yard on packed sand during the day and on deep sand during the night. The hoof block slowly wore down, and at 7 wk, it was placed back on concrete in the evening. At 8 wk, the block had completely worn off and the animal was no longer lame, but radiographs indicated minimal fracture healing. Radiographs performed at 7 mo indicated that there was still a radiolucent zone at the fracture line but calcification was evident at the margins of the fracture.


Porcelain gallbladder: relation between its type by ultrasound and incidence of cancer.
We report a case of porcelain gallbladder in a 67-year-old Japanese woman and review, summarize, and tabulate the literature describing its ultrasound appearance. We classify the ultrasound finding of porcelain gallbladder into two types: a complete type and an incomplete type. The relation between the type of porcelain gallbladder and pathological features is important because in the incomplete type the frequency of cancer was much higher than in the complete type.


Ubiquitin-conjugating enzyme UBE2C: molecular biology, role in tumorigenesis, and potential as a biomarker.
Ubiquitin-conjugating enzyme 2C (UBE2C) participates in cell cycle progression and checkpoint control by targeted degradation of short-lived proteins. As a conjugating enzyme, it directs polyubiquitination to preferred lysine in the substrates. In addition to its well-known role in cyclin B destruction that is essential for exit from mitosis, UBE2C also plays an important role in mitotic spindle checkpoint control. Cells overexpressing UBE2C ignore the mitotic spindle checkpoint signals and lose genomic stability, which is a hallmark of cancer. UBE2C expression is upregulated upon malignant transformation, and amplification of UBE2C is often seen at the chromosome level in cancers in a manner similar to c-Myc, which is directly upstream of UBE2C. UBE2C levels are upregulated in a wide range of solid tumors and hematological malignancies. The level of expression correlates with the aggressiveness of the tumor. High UBE2C expression is predictive of poor survival and perhaps high risk for relapse. UBE2C immunochemistry may be integrated into the diagnosis of thyroid malignancy and gliomas. This minireview summarizes what is known about the function of UBE2C focusing on its role in the regulation of spindle assembly checkpoint, its part in tumorigenesis, and its potential as a tumor marker for various cancers.