Litter of the hemiparasite Bartsia alpina enhances plant growth: evidence for a functional role in nutrient cycling.
Hemiparasitic angiosperms concentrate nutrients in their leaves and also produce high quality litter, which can decompose faster and release more nutrients than that of surrounding species. The impact of these litters on plant growth may be particularly important in nutrient-poor communities where hemiparisites can be abundant, such as the sub-Arctic. We tested the hypothesis that plant growth is enhanced by the litter of the hemiparasite Bartsia alpina, in comparison with litter of co-occurring dwarf shrub species, using a pot based bioassay approach. Growth of Betula nana and Poa alpina was up to 51% and 41% greater, respectively, in the presence of Bartsia alpina litter than when grown with dwarf shrub litter (Vaccinium uliginosum, Betula nana and Empetrum nigrum subsp. hermaphroditum). The nutrient concentrations of Betula nana plants grown with Bartsia alpina litter were almost double those of plants grown with dwarf shrub litter, and a significantly greater proportion of biomass was allocated to shoots rather than roots, strongly suggesting that nutrient availability was higher where Bartsia alpina litter was present. The presence of litter from dwarf shrubs, or the moss Hylocomium splendens, did not reduce the positive effect of Bartsia alpina litter on plant growth. E. nigrum litter did not appear to affect plant growth substantially differently from litter of other dwarf shrub species, despite earlier reports of its allelopathic action. The enhanced nutrient uptake and growth of plants in the presence of Bartsia alpina (and potentially other hemiparasitic species) litter could have important implications for communities in which it occurs, including enhanced survival of seedlings of co-occurring species and increased resource patchiness.


Localized cisplatin hyperpigmentation induced by pressure. A case report.
The spectrum of side effects induced by chemotherapy includes skin hyperpigmentation. This is prone to occur following treatment with alkylating agents and doxorubicin. More recently, hyperpigmentation was discovered in patients treated exclusively with intra-arterial cisplatin and was likely to develop over the dorsal surfaces of the hands and feet, elbows and knees, and operative incisions (trauma). We followed the clinical course of a patient with osteosarcoma treated with intravenous (i.v.) cisplatin, doxorubicin, and high-dose methotrexate. Following two courses of chemotherapy, hyperpigmentation developed along the sides of the thorax juxtaposed to the rubber shoulder pads of the patient's crutches. It appeared that the pigmentary change was due to localized pressure in the skin of the patient exposed to i.v. cisplatin. Factors associated with skin hyperpigmentation as a complication of chemotherapy are discussed. We believe that cisplatin was the major contributing factor. The mechanism for cisplatin-induced hyperpigmentation is undetermined. However, it has occurred in patients treated exclusively with intra-arterial cisplatin and can possibly be attributed to an effect on the melanocytes. / This is supported by experiences in which cisplatin extravasated into the tissues and resulted in a similar phenomenon. Reasons for excluding doxorubicin and methotrexate as causative agents are presented.


Hydrogen ameliorates oxidative stress via PI3K-Akt signaling pathway in UVB-induced HaCaT cells.
Chronic ultraviolet (UV) exposure-induced oxidative stress is associated with the pathogenesis of skin damage. However, the nuclear factor erythroid‑2‑related factor 2 (Nrf2) pathway is a critical factor in protecting cells against UVB‑induced injury through inhibiting oxidative stress. Furthermore, Nrf2 activation requires the involvement of the phosphoinositide-3 kinase (PI3K)/protein kinase B (AKT) pathway, which has a major role in survival of various cell types. Molecular hydrogen exerts protective effects on UV‑induced injury, but the underlying mechanisms have remained elusive. The present study assessed the protective effects of hydrogen against oxidative stress‑induced injury caused by UVB irradiation and investigated the molecular mechanisms. In vitro, UVB‑induced HaCaT cells were collected for the detection of reactive oxygen species, 8‑iso‑prostaglandin F2α, malondialdehyde via fluorescence spectrometry and ELISA; cell activity and cytotoxicity by MTT and lactate dehydrogenase assays, respectively. Additionally, the expression level of PI3K, Akt, Nrf2 and heme oxygenase‑1 (HO‑1) were investigated using western blot, etc. All of the results indicated that hydrogen decreased the levels of reactive oxygen species, 8‑iso‑prostaglandin F2α and malondialdehyde, and promoted the UVB exposure‑induced expression of PI3K, Akt, Nrf2 and heme oxygenase‑1 in HaCaT cells. Of note, PI3K inhibition partially reversed the effects of hydrogen on UVB‑induced HaCaT cells. Therefore, hydrogen effectively protects cells from UVB radiation‑induced oxidative stress by inhibiting Nrf2/HO‑1 activation through the PI3K/Akt signaling pathway.


Maternal diet as a risk factor for primary congenital glaucoma and defects of the anterior segment of the eye in the National Birth Defects Prevention Study.
Primary congenital glaucoma (PCG) and anterior segment defects (ASDs) are rare ocular malformations diagnosed early in life which can cause blindness. Pathogenic variants in several genes have been linked to these conditions, but little is known about nongenetic risk factors. We investigated the association between maternal nutrition and PCG and ASDs in the National Birth Defects Prevention Study, a large population-based, multicenter case-control study of major birth defects in the United States. Mothers of cases (n = 152) and control infants without a birth defect (n = 9,178) completed an interview which included a food frequency questionnaire capturing usual dietary intake in the year before pregnancy. Maternal nutrition was assessed through individual nutrient intake, calculating a Diet Quality Index for Pregnancy (DQI-P) score for each mother, and using latent class analysis to empirically derive four dietary patterns. We calculated adjusted odds ratios (aORs) and 95% confidence intervals (CI) using logistic regression. The results for individual nutrients varied, with some having an inverse or U-shaped pattern of association with increasing intake. The DQI-P was not associated with risk of PCG and ASDs (aOR 0.91; CI 0.49-1.66, highest vs. lowest quartile). The dietary pattern analysis suggested lower odds among women with a Prudent and Mexican dietary pattern (aOR 0.82, 95% CI 0.52-1.29; aOR 0.80, 95% CI 0.36-1.78, respectively) compared to those with a Western dietary pattern. We found that higher intake of some nutrients and certain dietary patterns may be inversely associated with PCG and ASDs, though caution is urged due to imprecision of estimates.


Determination of potentially novel compensatory mutations in rpoc associated with rifampin resistance and rpob mutations in Mycobacterium tuberculosis Clinical isolates from peru.
Rifampicin (RIF) resistance in Mycobacterium tuberculosis is frequently caused by mutations in the rpoB gene. These mutations are associated with a fitness cost, which can be overcome by compensatory mutations in other genes, among which rpoC may be the most important. We analyzed 469 Peruvian M. tuberculosis clinical isolates to identify compensatory mutations in rpoC/rpoA associated with RIF resistance. The M. tuberculosis isolates were collected and tested for RIF susceptibility and spoligotyping. Samples were sequenced and aligned to the reference genome to identify mutations. By analyzing the sequences and the metadata, we identified a list of rpoC mutations exclusively associated with RIF resistance and mutations in rpoB. We then evaluated the distribution of these mutations along the protein sequence and tridimensional structure. One hundred and twenty-five strains were RIF susceptible and 346 were resistant. We identified 35 potential new compensatory mutations, some of which were distributed on the interface surface between rpoB and rpoC, arising in clusters and suggesting the presence of hotspots for compensatory mutations. This study identifies 35 putative novel compensatory mutations in the β' subunit of M. tuberculosis RNApol. Six of these (S428T, L507V, A734V, I997V, and V1252LM) are considered most likely to have a compensatory role, as they fall in the interaction zone of the two subunits and the mutation did not lead to any change in the protein's physical-chemical properties.


In Situ Electron Microscopy of Plasmon-Mediated Nanocrystal Synthesis.
Chemical processes driven by nonthermal energy (e.g., visible light) are attractive for future approaches to energy conversion, synthesis, photocatalysis, and so forth. The growth of anisotropic metal nanostructures mediated by excitation of a localized surface plasmon resonance (LSPR) is a prototype example of such a reaction. Important aspects, notably the growth mechanism and a possible role of plasmonic "hot spots" within the metal nanostructures, remain poorly understood. Here, we use in situ electron microscopy to stimulate and image the plasmon-mediated growth of triangular Ag nanoprisms in solution. The quantification of the time-dependent evolution of the lateral size and thickness of the nanoprisms, enabled by nanometer-scale real-time microscopy in solution, shows a transition from an early stage of uniform Ag0 incorporation exclusively in the prism side facets to a size regime with accelerated growth in thickness. Differences in attachment rate at this advanced stage correlate with local plasmonic field enhancements, which allows determining the range over which charge carriers transferred from plasmonic hot spots can drive chemistry. Such data support the development of nonthermal chemical processes that depend on plasmonic light harvesting and the transfer of nonequilibrium charge carriers.


Regulation of endothelial-specific transgene expression by the LacI repressor protein in vivo.
Genetically modified mice have played an important part in elucidating gene function in vivo. However, conclusions from transgenic studies may be compromised by complications arising from the site of transgene integration into the genome and, in inducible systems, the non-innocuous nature of inducer molecules. The aim of the present study was to use the vascular system to validate a technique based on the bacterial lac operon system, in which transgene expression can be repressed and de-repressed by an innocuous lactose analogue, IPTG. We have modified an endothelium specific promoter (TIE2) with synthetic LacO sequences and made transgenic mouse lines with this modified promoter driving expression of mutant forms of connexin40 and an independently translated reporter, EGFP. We show that tissue specificity of this modified promoter is retained in the vasculature of transgenic mice in spite of the presence of LacO sequences, and that transgene expression is uniform throughout the endothelium of a range of adult systemic and cerebral arteries and arterioles. Moreover, transgene expression can be consistently down-regulated by crossing the transgenic mice with mice expressing an inhibitor protein LacI(R), and in one transgenic line, transgene expression could be de-repressed rapidly by the innocuous inducer, IPTG. We conclude that the modified bacterial lac operon system can be used successfully to validate transgenic phenotypes through a simple breeding schedule with mice homozygous for the LacI(R) protein.


[Local invasive treatment of chronic pain].
The literature on methods of invasive local treatment of chronic pain was analyzed. We reviewed 14 publications including meta-analyses and systematic reviews. The use of regional anesthesia conducted by anesthesiologists in pain clinics demonstrated the evidence based efficacy of different types of peridural injections of local anesthetics with steroids in patients with root pain syndromes at cervical and lumbar levels. Therapeutic blockades of the occipital nerve is effective method of treatment of cervicogenic and cluster headache as well as occipital nerve neuralgia. There are clear indications of the efficacy of local injections in primary chronic cephalgia (migraine and headache of tension). The possibility of the abortion of the pain information flow in peripheral nociceptive pathways and, as a consequence, breaking the vicious circle is emphasized. Issues on the efficacy of local injections at trigger points in the treatment of chronic pain are highlighted.


Triple-Negative Breast Cancer Comparison With Canine Mammary Tumors From Light Microscopy to Molecular Pathology.
Many similar characteristics in human and dog cancers including, spontaneous development, clinical presentation, tumor heterogeneity, disease progression, and response to standard therapies have promoted the approval of this comparative model as an alternative to mice. Breast cancer represents the second most frequent neoplasm in humans after lung cancer. Triple-negative breast cancers (TNBC) constitute around 15% of all cases of breast cancer and do not express estrogen receptor (ER), progesterone receptor (PR), and do not overexpress human epidermal growth factor receptor 2 (HER2). As a result, they do not benefit from hormonal or trastuzumab-based therapy. Patients with TNBC have worse overall survival than patients with non-TNBC. Lehmann and collaborators described six different molecular subtypes of TNBC which further demonstrated its transcriptional heterogeneity. This six TNBC subtype classification has therapeutic implications. Breast cancer is the second most frequent neoplasm in sexually intact female dogs after skin cancer. Canine mammary tumors are a naturally occurring heterogeneous group of cancers that have several features in common with human breast cancer (HBC). These similarities include etiology, signaling pathway activation, and histological classification. Molecularly CMTs are more like TNBCs, and therefore dogs are powerful spontaneous models of cancer to test new therapeutic approaches, particularly for human TNBCs. More malignant tumors of the breast are more often ER and PR negative in both humans and dogs. Promising breast cancer biomarkers in both humans and canines are cancer-associated stroma (CAS), circulating tumor cells and tumor DNA (ctDNA), exosomes and miRNAs, and metabolites.


Choline: Exploring the Growing Science on Its Benefits for Moms and Babies.
The importance of ensuring adequate choline intakes during pregnancy is increasingly recognized. Choline is critical for a number of physiological processes during the prenatal period with roles in membrane biosynthesis and tissue expansion, neurotransmission and brain development, and methyl group donation and gene expression. Studies in animals and humans have shown that supplementing the maternal diet with additional choline improves several pregnancy outcomes and protects against certain neural and metabolic insults. Most pregnant women in the U.S. are not achieving choline intake recommendations of 450 mg/day and would likely benefit from boosting their choline intakes through dietary and/or supplemental approaches.


Reactivity control of C-H bond activation over vanadium-silver bimetallic oxide cluster cations.
Vanadium-silver bimetallic oxide cluster ions (V(x)Ag(y)O(z)(+); x=1-4, y=1-4, z=3-11) are produced by laser ablation and reacted with ethane in a fast-flow reactor. A reflectron time of flight (Re-TOF) mass spectrometer is used to detect the cluster distribution before and after the reactions. Hydrogen atom abstraction (HAA) reactions are identified over VAgO(3)(+), V(2)Ag(2)O(6)(+), V(2)Ag(4)O(7)(+), V(3)AgO(8)(+), V(3)Ag(3)O(9)(+), and V(4)Ag(2)O(11)(+) ions, in which the oxygen-centered radicals terminally bonded on V atoms are active sites for the facile HAA reactions. DFT calculations are performed to study the structures, bonding, and reactivity. The reaction mechanisms of V(2)Ag(2)O(6)(+) +C(2)H(6) are also given. The doped Ag atoms with a valence state of +1 are highly dispersed at the periphery of the V(x)Ag(y)O(z)(+) cluster ions. The reactivity can be well-tuned gradually by controlling the number of Ag atoms. The steric protection due to the peripherally bonded Ag atoms greatly enhances the selectivity of the V-Ag bimetallic oxide clusters with respect to the corresponding pure vanadium oxide systems.


Impact of a New York City Supportive Housing Program on Housing Stability and Preventable Health Care among Homeless Families.
To assess the impact of a New York City supportive housing program on housing stability and preventable emergency department (ED) visits/hospitalizations among heads of homeless families with mental and physical health conditions or substance use disorders. Multiple administrative data from New York City and New York State for 966 heads of families eligible for the program during 2007-12. We captured housing events and health care service utilization during 2 years prior to the first program eligibility date (baseline) and 2 years postbaseline. We performed sequence analysis to measure housing stability and compared housing stability and preventable ED visits and hospitalizations between program participants (treatment group) and eligible applicants not placed in the program (comparison group) via marginal structural modeling. We matched electronically collected data. Eighty-seven percent of supportive housing tenants experienced housing stability in 2 years postbaseline. Compared with unstably housed heads of families in the comparison group, those in the treatment group were 0.60 times as likely to make preventable ED visits postbaseline (95% CI = 0.38, 0.96). Supportive housing placement was associated with improved housing stability and reduced preventable health care visits among homeless families.


Community based intervention to optimize osteoporosis management: randomized controlled trial.
Osteoporosis-related fractures are a significant public health concern. Interventions that increase detection and treatment of osteoporosis are underutilized. This pragmatic randomised study was done to evaluate the impact of a multifaceted community-based care program aimed at optimizing evidence-based management in patients at risk for osteoporosis and fractures. This was a 12-month randomized trial performed in Ontario, Canada. Eligible patients were community-dwelling, aged ≥55 years, and identified to be at risk for osteoporosis-related fractures. Two hundred and one patients were allocated to the intervention group or to usual care. Components of the intervention were directed towards primary care physicians and patients and included facilitated bone mineral density testing, patient education and patient-specific recommendations for osteoporosis treatment. The primary outcome was the implementation of appropriate osteoporosis management. 101 patients were allocated to intervention and 100 to control. Mean age of participants was 71.9 ± 7.2 years and 94% were women. Pharmacological treatment (alendronate, risedronate, or raloxifene) for osteoporosis was increased by 29% compared to usual care (56% [29/52] vs. 27% [16/60]; relative risk [RR] 2.09, 95% confidence interval [CI] 1.29 to 3.40). More individuals in the intervention group were taking calcium (54% [54/101] vs. 20% [20/100]; RR 2.67, 95% CI 1.74 to 4.12) and vitamin D (33% [33/101] vs. 20% [20/100]; RR 1.63, 95% CI 1.01 to 2.65). A multi-faceted community-based intervention improved management of osteoporosis in high risk patients compared with usual care. This trial has been registered with clinicaltrials.gov (ID: NCT00465387).


Treatment of Infections by OXA-48-Producing Enterobacteriaceae.
Carbapenemase-producing Enterobacteriaceae (CPE) contribute significantly to the global public health threat of antimicrobial resistance. OXA-48 and its variants are unique carbapenemases with low-level hydrolytic activity toward carbapenems but no intrinsic activity against expanded-spectrum cephalosporins. blaOXA-48 is typically located on a plasmid but may also be integrated chromosomally, and this gene has progressively disseminated throughout Europe and the Middle East. Despite the inability of OXA-48-like carbapenemases to hydrolyze expanded-spectrum cephalosporins, pooled isolates demonstrate high variable resistance to ceftazidime and cefepime, likely representing high rates of extended-spectrum beta-lactamase (ESBL) coproduction. In vitro data from pooled studies suggest that avibactam is the most potent beta-lactamase inhibitor when combined with ceftazidime, cefepime, aztreonam, meropenem, or imipenem. Resistance to novel avibactam combinations such as imipenem-avibactam or aztreonam-avibactam has not yet been reported in OXA-48 producers, although only a few clinical isolates have been tested. Although combination therapy is thought to improve the chances of clinical cure and survival in CPE infection, successful outcomes were seen in ∼70% of patients with infections caused by OXA-48-producing Enterobacteriaceae treated with ceftazidime-avibactam monotherapy. A carbapenem in combination with either amikacin or colistin has achieved treatment success in a few case reports. Uncertainty remains regarding the best treatment options and strategies for managing these infections. Newly available antibiotics such as ceftazidime-avibactam show promise; however, recent reports of resistance are concerning. Newer choices of antimicrobial agents will likely be required to combat this problem.


TLR3 deficiency renders astrocytes permissive to herpes simplex virus infection and facilitates establishment of CNS infection in mice.
Herpes simplex viruses (HSVs) are highly prevalent neurotropic viruses. While they can replicate lytically in cells of the epithelial lineage, causing lesions on mucocutaneous surfaces, HSVs also establish latent infections in neurons, which act as reservoirs of virus for subsequent reactivation events. Immunological control of HSV involves activation of innate immune pattern-recognition receptors such as TLR3, which detects double-stranded RNA and induces type I IFN expression. Humans with defects in the TLR3/IFN pathway have an elevated susceptibility to HSV infections of the CNS. However, it is not known what cell type mediates the role of TLR3 in the immunological control of HSV, and it is not known whether TLR3 sensing occurs prior to or after CNS entry. Here, we show that in mice TLR3 provides early control of HSV-2 infection immediately after entry into the CNS by mediating type I IFN responses in astrocytes. Tlr3-/- mice were hypersusceptible to HSV-2 infection in the CNS after vaginal inoculation. HSV-2 exhibited broader neurotropism in Tlr3-/- mice than it did in WT mice, with astrocytes being most abundantly infected. Tlr3-/- mice did not exhibit a global defect in innate immune responses to HSV, but astrocytes were defective in HSV-induced type I IFN production. Thus, TLR3 acts in astrocytes to sense HSV-2 infection immediately after entry into the CNS, possibly preventing HSV from spreading beyond the neurons mediating entry into the CNS.


Outcome-driven thresholds for ambulatory pulse pressure in 9938 participants recruited from 11 populations.
Evidence-based thresholds for risk stratification based on pulse pressure (PP) are currently unavailable. To derive outcome-driven thresholds for the 24-hour ambulatory PP, we analyzed 9938 participants randomly recruited from 11 populations (47.3% women). After age stratification (<60 versus ≥60 years) and using average risk as reference, we computed multivariable-adjusted hazard ratios (HRs) to assess risk by tenths of the PP distribution or risk associated with stepwise increasing (+1 mm Hg) PP levels. All adjustments included mean arterial pressure. Among 6028 younger participants (68 853 person-years), the risk of cardiovascular (HR, 1.58; P=0.011) or cardiac (HR, 1.52; P=0.056) events increased only in the top PP tenth (mean, 60.6 mm Hg). Using stepwise increasing PP levels, the lower boundary of the 95% confidence interval of the successive thresholds did not cross unity. Among 3910 older participants (39 923 person-years), risk increased (P≤0.028) in the top PP tenth (mean, 76.1 mm Hg). HRs were 1.30 and 1.62 for total and cardiovascular mortality, and 1.52, 1.69, and 1.40 for all cardiovascular, cardiac, and cerebrovascular events. The lower boundary of the 95% confidence interval of the HRs associated with stepwise increasing PP levels crossed unity at 64 mm Hg. While accounting for all covariables, the top tenth of PP contributed less than 0.3% (generalized R(2) statistic) to the overall risk among the elderly. Thus, in randomly recruited people, ambulatory PP does not add to risk stratification below age 60; in the elderly, PP is a weak risk factor with levels below 64 mm Hg probably being innocuous.


Impacts of care-giving and sources of support: a comparison of end-of-life and non-end-of-life caregivers in Canada.
This is the second in a series of papers that deal with care-giving in Canada, as based on data available from the Canadian General Social Survey (2007). Building on the first paper, which reviewed the differences between short-term, long-term and end-of-life (EOL) caregivers, this paper uniquely examines the caregiver supports employed by EOL caregivers when compared to non-EOL caregivers (short-term and long-term caregivers combined). Both papers employ data from Statistics Canada's General Social Survey (GSS Cycle 21: 2007). The GSS includes three modules, where respondents were asked questions about the unpaid home care assistance that they had provided in the last 12 months to someone at EOL or with either a long-term health condition or a physical limitation. The objective of this research paper was to investigate the link between the impact of the care-giving experience and the caregiver supports received, while also examining the differences in these across EOL and non-EOL caregivers. By way of factor analysis and regression modelling, we examine differences between two types of caregivers: (i) EOL and (ii) non-EOL caregivers. The study revealed that with respect to socio-demographic characteristics, health outcomes and caregiver supports, EOL caregivers were consistently worse off. This suggests that although all non-EOL caregivers are experiencing negative impacts from their care-giving role, comparatively greater supports are needed for EOL caregivers.


Long-term outcomes after laparoscopy-assisted gastrectomy for advanced gastric cancer: a large-scale multicenter retrospective study.
Recently, the number of laparoscopic procedures for gastric cancer has increased rapidly. Laparoscopic surgery is reported to have many advantages over open gastrectomy with oncologic safety in early gastric cancer. However, there were few reports on long-term outcomes of laparoscopy-assisted gastrectomy (LAG) for advanced gastric cancer (AGC). The aim of this study was to investigate long-term survival outcomes after LAG for AGC. The data of 1,485 patients who underwent LAG between April 1998 and December 2005 by ten surgeons at ten hospitals were collected retrospectively. Among them, 239 patients who were diagnosed with AGC on final pathologic examination were enrolled in the present study to investigate long-term clinical outcomes. The ratio of male to female patients was 151:88 and the mean age was 57.1 years. One hundred ninety-three subtotal gastrectomies, 41 total gastrectomies, and 5 proximal gastrectomies were performed. D1 + α, D1 + β, and D2 lymph node dissections were performed for 14, 62, and 163 cases, respectively. The median follow-up period was 55.4 months. The overall 5-year survival rate of the 239 AGC patients was 78.8% and the disease-specific 5-year survival rate was 85.6%. The 5-year survival rates of the TNM staging system's (7th ed.) stages were 90.5% (stage Ib, n = 86), 86.4% (stage IIa, n = 53), 78.3% (stage IIb, n = 44), 52.8% (stage IIIa, n = 24), 52.9% (stage IIIb, n = 24), and 37.5% (stage IIIc, n = 8) (p < 0.001). The long-term survival outcome rates of LAG for AGC in the present study were comparable to those previously reported for open gastrectomy. Based on the present results, a well-designed phase III trial comparing LAG and open gastrectomy for AGC will be needed to affirm the validity of LAG for AGC.


Generally Applicable Transformation Protocols for Fluorescent Nanodiamond Internalization into Cells.
Fluorescent nanodiamonds (FNDs) are promising nanoprobes, owing to their stable and magnetosensitive fluorescence. Therefore they can probe properties as magnetic resonances, pressure, temperature or strain. The unprecedented sensitivity of diamond defects can detect the faint magnetic resonance of a single electron or even a few nuclear spins. However, these sensitivities are only achieved if the diamond probe is close to the molecules that need to be detected. In order to utilize its full potential for biological applications, the diamond particle has to enter the cell. Some model systems, like HeLa cells, readily ingest particles. However, most cells do not show this behavior. In this article we show for the first time generally applicable methods, which are able to transport fluorescent nanodiamonds into cells with a thick cell wall. Yeast cells, in particular Saccharomyces cerevisiae, are a favored model organism to study intracellular processes including aging on a cellular level. In order to introduce FNDs in these cells, we evaluated electrical transformation and conditions of chemical permeabilization for uptake efficiency and viability. 5% DMSO (dimethyl sulfoxide) in combination with optimized chemical transformation mix leads to high uptake efficiency in combination with low impact on cell biology. We have evaluated all steps in the procedure.


Estimating the exposure of small mammals at three sites within the Chernobyl exclusion zone--a test application of the ERICA Tool.
An essential step in the development of any modelling tool is the validation of its predictions. This paper describes a study conducted within the Chernobyl exclusion zone to acquire data to conduct an independent test of the predictions of the ERICA Tool which is designed for use in assessments of radiological risk to the environment. Small mammals were repeatedly trapped at three woodland sites between early July and mid-August 2005. Thermoluminescent dosimeters mounted on collars were fitted to Apodemus flavicollis, Clethrionomys glareolus and Microtus spp. to provide measurements of external dose rate. A total of 85 TLDs were recovered. All animals from which TLDs were recovered were live-monitored to determine (90)Sr and (137)Cs whole-body activity concentrations. A limited number of animals were also analysed to determine (239,240)Pu activity concentrations. Measurements of whole-body activity concentrations and dose rates recorded by the TLDs were compared to predictions of the ERICA-Tool. The predicted (90)Sr and (137)Cs mean activity concentrations were within an order of magnitude of the observed data means. Whilst there was some variation between sites in the agreement between measurements and predictions this was consistent with what would be expected from the differences in soil types at the sites. Given the uncertainties of conducting a study such as this, the agreement observed between the TLD results and the predicted external dose rates gives confidence to the predictions of the ERICA Tool.


Extent of low-grade dysplasia is a risk factor for the development of esophageal adenocarcinoma in Barrett's esophagus.
Previous studies that evaluated extent of high-grade dysplasia (HGD) as a risk factor for esophageal adenocarcinoma (EA) in Barrett's esophagus (BE) were conflicting, and no prior study has evaluated extent of low-grade dysplasia (LGD) as a risk factor. The aim of this discovery study was to evaluate the hypothesis that extent of LGD and HGD are risk factors for progression to EA. We evaluated baseline biopsies from 77 BE patients with dysplasia including 44 who progressed to EA and 33 who did not progress during follow-up. The total numbers of LGD and HGD crypts were determined separately by counting all crypts and the extent of LGD, HGD, and total dysplasia were correlated with EA outcome. Thirty-one and 46 patients had a maximum diagnosis of LGD and HGD, respectively. When the crypts were stratified by dysplasia grade, the mean number of LGD crypts per patient was borderline higher in progressors (93.9) compared with nonprogressors (41.2, P= 0.07), and the mean proportion of LGD crypts per patient was significantly higher in progressors (46.4%vs 26.0%, P= 0.037). Neither the mean number of HGD crypts per patient (P= 0.14) nor the mean proportion of HGD crypts per patient (P= 0.20) was significantly associated with EA outcome. The extent of LGD is a significant risk factor for the development of EA in BE in this study. Although the presence of HGD is significantly associated with a greater relative risk for development of EA, the extent of HGD was not an independent risk factor for progression.


[Lymphoma in patients who have undergone organ transplantation: severe and variable clinical presentation].
Two patients presented with post-transplant lymphoproliferative disorder (PTLD). PTLD encompasses a broad range ofoften malignant proliferations of lymphoid tissue arising in the immunocompromised host after transplantation. The first patient, a 62-year-old woman, received a bilateral lung transplant due to end-stage emphysema and was diagnosed with PTLD 27 days after transplantation. Treatment consisted of reduction in immunosuppression and administration of rituximab. The PTLD regressed. The second patient, a 57-year-old woman, presented with a massively disseminated PTLD 12 years after kidney transplantation. Immunosuppression was reduced and rituximab was administered, but no response was observed. Despite salvage chemotherapy, the patient died due to progressive disease. These two cases illustrate the heterogeneous presentation of PTLD. The condition is caused by the proliferation of B lymphocytes infected with Epstein-Barr virus (EBV) that are no longer controlled by EBV-specific cytotoxic T lymphocytes, due to the immunosuppressive medication given to prevent transplant rejection. Regression of the lymphoma may be achieved by reducing the immunosuppression or treating with rituximab, which attacks B lymphocytes.


A new crystal form of mouse thiamin pyrophosphokinase.
Thiamin pyrophosphokinase (TPK) transfers a pyrophosphate group from ATP to the hydroxyl group of thiamin and produces thiamin pyrophosphate (TPP). TPP is the cofactor of metabolically important enzymes such as pyruvate dehydrogenase, α-ketoglutarate dehydrogenase, branched-chain α-keto acid dehydrogenase, transketolase and 2-hydroxyphytanoyl-CoA lyase. Thiamin deficiency results in Wernike-Korsakof Syndrome (WKS) due to neurological disorder and wet beriberi, a potentially fatal cardiovascular disease. Mouse TPK associates as a dimer revealed by previous solved crystallographic structures. In this study, we report mouse TPK complexed with TPP-Mg(2+) and thiamin -Mg(2+), respectively, in a new crystal form. In these two structures, four mouse TPK molecules were found in each asymmetric unit. Although we cannot rule out this tetramer form can be an artifact from crystal packing, mouse TPK tetramer has a more closed ATP binding pocket and has the potential to provide specific interactions between mouse TPK and ATP compared with the previous dimeric structure and is likely to be an active form.


Uterine arterial vasoconstrictions mediated by ovarian nerves in virgin and postpartum rats.
In most mammals, including humans, pregnancy results in the loss of most uterine vasomotor fibers. These experiments determined whether, despite this denervation, sympathetic nerves mediated uterine vasoconstrictions in the rat 24 h after delivery. Both virgin and uniparous postpartum rats were anesthetized with urethan. Femoral vessels were cannulated for measurement of arterial pressure and intravenous administration of fluids and drugs. Blood flow was measured in a uterine artery after ligation of all anastomotic ovarian vessels. Electrical stimulation of ovarian nerve efferents elicited frequency-dependent uterine vasoconstrictions in both virgin and postpartum rats. Vasoconstrictions in postpartum rats were not significantly different from those observed in virgins. In both virgin and postpartum rats, neurogenic vasoconstrictions were reduced by combined alpha 1- and alpha 2-adrenergic blockade. We conclude that the uterine branches of the ovarian nerve mediate adrenergic uterine vasoconstrictions. In the largely denervated uterus of the postpartum rat, these vasoconstrictions may be mediated by surviving innervation of the uterine artery and its major branches. Sympathetic vasoconstriction acting at these sites would constitute an effective defense against postpartum hemorrhage.


Acute episodes of predator exposure in conjunction with chronic social instability as an animal model of post-traumatic stress disorder.
People who are exposed to horrific, life-threatening experiences are at risk for developing post-traumatic stress disorder (PTSD). Some of the symptoms of PTSD include persistent anxiety, exaggerated startle, cognitive impairments and increased sensitivity to yohimbine, an alpha(2)-adrenergic receptor antagonist. We have taken into account the conditions known to induce PTSD, as well as factors responsible for long-term maintenance of the disorder, to develop an animal model of PTSD. Adult male Sprague-Dawley rats were administered a total of 31 days of psychosocial stress, composed of acute and chronic components. The acute component was a 1-h stress session (immobilization during cat exposure), which occurred on Days 1 and 11. The chronic component was that on all 31 days the rats were given unstable housing conditions. We found that psychosocially stressed rats had reduced growth rate, reduced thymus weight, increased adrenal gland weight, increased anxiety, an exaggerated startle response, cognitive impairments, greater cardiovascular and corticosterone reactivity to an acute stressor and heightened responsivity to yohimbine. This work demonstrates the effectiveness of acute inescapable episodes of predator exposure administered in conjunction with daily social instability as an animal model of PTSD.


Design, Synthesis, and Biological Evaluation of Trisubstituted Piperazine Derivatives as Noncovalent Severe Acute Respiratory Syndrome Coronavirus 2 Main Protease Inhibitors with Improved Antiviral Activity and Favorable Druggability.
The ongoing transmission of SARS-CoV-2 necessitates the development of additional potent antiviral agents capable of combating the current highly infectious variants and future coronaviruses. Here, we present the discovery of potent nonpeptide main protease (Mpro) inhibitors with prominent antiviral activity and improved pharmacokinetic properties. Three series of 1,2,4-trisubstituted piperazine derivatives were designed and synthesized, and the optimal GC-78-HCl demonstrated high enzyme-inhibitory potency (IC50 = 0.19 μM) and exhibited excellent antiviral activity (EC50 = 0.40 μM), reaching the same level as Nirmatrelvir (EC50 = 0.38 μM). Additionally, GC-78-HCl displayed potent antiviral activities against various SARS-CoV-2 variants as well as HCoV-OC43 and HCoV-229E, indicating its potential broad-spectrum anticoronaviral activity. Notably, the pharmacokinetic properties of GC-78-HCl were somewhat enhanced compared to those of the lead compound. Furthermore, the cocrystal and molecular docking elucidated the mechanism of action. In conclusion, we discovered a novel nonpeptidic Mpro inhibitor with promising antiviral activity and a favorable pharmacokinetic profile.


Inhibitory action of oxytocin on spontaneous contraction of rat distal colon by nitrergic mechanism: involvement of cyclic GMP and apamin-sensitive K+ channels.
The mechanisms underlying the inhibitory effects of oxytocin (OT) on colon tone are not totally understood. We explore the mechanisms of OT on spontaneous contractility in rat distal colon and identify the mediators involved in this action. In rat distal colon strips, mechanical activity was analysed and the production of nitric oxide (NO) in tissue loaded with the fluorochrome DAF-FM was visualized by confocal microscopy. OT receptor (OTR) expression was determined by Western blotting and immunofluorescence. In rat distal colon, OT produced a concentration-dependent reduction in the spontaneous contraction, which was abolished by the OTR antagonist atosiban, the neural blocker tetrodotoxin and the inhibitor of neuronal nitric oxide synthase (nNOS) NPLA. The inhibitory effects of OT were not affected by propranolol, atropine, the nicotinic cholinoceptor blocker hexamethonium, the vasoactive intestinal peptide receptor antagonist VIPHyb, the P2 purinoceptor antagonist PPADS, the adenosine A1 receptors antagonist DPCPX and the prostacyclin receptor antagonist Ro1138452. The soluble guanylyl cyclase (sGC) inhibitor ODQ and the small conductance Ca2+ -activated K+ (Ca K+ ) channels blocker apamin significantly reduced the relaxation induced by OT, nicotine, sodium nitroprusside and the sGC activator BAY 41-2272. The neural release of NO elicited by OT was prevented by NPLA, tetrodotoxin and atosiban. The presence of the OTR and its co-localization with nNOS was detected by immunohistochemistry and Western blotting experiments. These results demonstrate the NO release from enteric neurones induced by activation of OTR mediates distal colon relaxation. sGC and small conductance Ca K+ channels are involved in this relaxation.


Metagenomics, gene discovery and the ideal biocatalyst.
With the rapid development of powerful protein evolution and enzyme-screening technologies, there is a growing belief that optimum conditions for biotransformation processes can be established without the constraints of the properties of the biocatalyst. These technologies can then be applied to find the "ideal biocatalyst" for the process. In identifying the ideal biocatalyst, the processes of gene discovery and enzyme evolution play major roles. However, in order to expand the pool genes for in vitro evolution, new technologies, which circumvent the limitations of microbial culturability, must be applied. These technologies, which currently include metagenomic library screening, gene-specific amplification methods and even full metagenomic sequencing, provide access to a volume of "sequence space" that is not addressed by traditional screening.


Low serum 25-hydroxyvitamin D concentrations are associated with increased likelihood of having depressive symptoms among Japanese workers.
Accumulating evidence suggests a protective role of vitamin D against mood disorders; however, epidemiologic studies are scarce in working populations. We investigated cross-sectionally the association of serum vitamin D status and depressive symptoms among Japanese workers. Participants were 1786 employees (9% women), aged 19-69 y, who received health check-ups and participated in a nutrition and health survey. Serum 25-hydroxyvitamin D [25(OH)D] concentrations were measured with the use of a competitive protein binding assay. Depressive symptoms were assessed by using the Center for Epidemiologic Studies Depression (CES-D) scale. Logistic regression was used to estimate ORs with adjustment for potential confounding variables including dietary factors. Overall, 92% of study participants had suboptimal vitamin D status [25(OH)D <30 μg/L]. Depressive symptoms were inversely associated with 25(OH)D. Compared with those with a 25(OH)D concentration of <20 μg/L, multivariable-adjusted ORs (95% CIs) for depressive symptoms (CES-D scale score ≥16) were 0.75 (0.59, 0.95) and 0.66 (0.41, 1.06) for those with a 25(OH)D concentration of 20-29 μg/L and ≥30 μg/L, respectively (P-trend = 0.01). After further adjustment for leisure-time physical activity and shift work (factors closely related to photo-initiated vitamin D production), the OR (95% CI) for the highest category of 25(OH)D was 0.70 (0.43, 1.14). The association between 25(OH)D and depressive symptoms appears to be linear, according to restricted cubic spline regression. Results suggest that lower concentrations of circulating vitamin D are associated with increased likelihood of having depressive symptoms among apparently healthy workers.


Malignancies in Korean patients with immunoglobulin G4-related disease.
Several studies have implicated increased risk of malignancy in patients with immunoglobulin G4-related disease (IgG4-RD). Hence, we first evaluated the risk and the type of malignancy in Korean IgG4-RD patients. Clinical and laboratory results of patients fulfilling the comprehensive diagnostic criteria for IgG4-RD were retrospectively collected between January 2006 and November 2015. One hundred and eighteen patients were included, and 35 were classified as having definite, 83 as possible/probable IgG4-RD. We compared baseline clinical and laboratory variables between patients with definite and those with possible/probable IgG4-RD and calculated standardized incidence ratios (SIRs) for malignancies. The median age was 60 years old and 91 patients (77.1%) were male. Malignancies were found in 12 of 118 patients (10.1%), and lymphoma (4, 25.0%) was the most frequent malignancy related to IgG4-RD. Definite IgG4-RD patients had higher serum IgG levels and frequent multiple organ involvement. The overall incidence of malignancy was increased in patients with IgG4-RD compared to the general population (SIR 23.08 [95% CI 11.92-40.31]), and the overall risk of SIRs was highest in non-Hodgkin lymphoma (SIR 400.00 [95% CI 109.00-1024.00]). Our data showed that increased risk of malignancy, especially lymphomas, was observed in Korean IgG4-RD patients.


[Prevalence of clinical and subclinical forms of anorexia and bulimia nervosa among working females and males].
In a recent multicentre study it turned out that both clinical and subclinical eating disorders are more prevalent in Hungarian college students than among their Austrian and German counterparts. Now the prevalence of eating disorders was assessed in a population of Hungarian workers. From 1800 working females and males 762 returned the questionnaires; data of 689 subjects (571 females and 118 males) were analysed. Among females the prevalence rates of bulimia nervosa (according to DSM-III-R), subclinical bulimia nervosa and subclinical anorexia nervosa are 0.7% (n = 4), 5.3% (n = 30) and 0.7% (n = 4), respectively. In males two persons (1.7%) met DSM-III-R criteria for bulimia nervosa, three (2.5%) were identified as having subclinical bulimia nervosa and two (1.7%) subclinical anorexia nervosa. No case of clinical anorexia nervosa was found. So the overall prevalences of clinical and subclinical eating disorder syndromes are 6.7% for females and 5.9% for males in the working population. These disorders that mostly remain hidden in the population under study do not seem to be quite recent. These rather unfavorably high prevalence rates can be explained by the tendency for over-identification with Western norms and values; in addition, in the group of workers, the risk factors are supposed to be combined with the relative lack of protective factors.


Sugar-sweetened Beverage Consumption Among U.S. Youth, 2011-2014.
Data from the National Health and Nutrition Examination Survey •Almost two-thirds of boys and girls consumed at least one sugar-sweetened beverage on a given day. •Boys consumed an average 164 kilocalories (kcal) from sugar-sweetened beverages, which contributed 7.3% of total daily caloric intake. Girls consumed an average 121 kcal from sugar-sweetened beverages, which contributed 7.2% of total daily caloric intake. •Among both boys and girls, older youth had the highest mean intake and percentage of daily calories from sugar-sweetened beverages relative to younger children. •Non-Hispanic Asian boys and girls consumed the least calories and the lowest percentage of total calories from sugar-sweetened beverages compared with non-Hispanic white, non-Hispanic black, and Hispanic boys and girls. Sugar-sweetened beverages contribute calories and added sugars to the diets of U.S. children (1). Studies have suggested a link between the consumption of sugar-sweetened beverages and dental caries, weight gain, type 2 diabetes, dyslipidemia, and nonalcoholic fatty liver disease in children (2-6). The 2015-2020 Dietary Guidelines for Americans recommend reducing added sugars consumption to less than 10% of calories per day and, specifically, to choose beverages with no added sugars (1). This report presents results for consumption of sugar-sweetened beverages among U.S. youth aged 2-19 years for 2011-2014 by sex, age, and race and Hispanic origin.


Determinants of ignition times: Topographies of cell assemblies and the activation delays they imply.
The cell assembly model of language posits that words are laid down in the cortex by discrete sets of neurons distributed over specific parts of the brain. The strong internal links of these "word webs" may not only bind articulatory and acoustic knowledge of a lexical item, they may also link word and meaning; for example, by connecting neuron populations related to word forms to those of actions and perceptions to which the words refer. Therefore, the cortical activation elicited by words should reflect aspects of word meaning, a postulate that has received strong support from recent work using neurophysiological and metabolic imaging. Segalowitz & Lane make the point that this neurobiological model can also be used to predict reaction times in behavioral experiments, using the behavioral distinction between content and function words as an example. We acclaim their view, but warn that response times might be related to different mechanisms at the neuronal level, including the cortical distribution and internal connectivity of cell assemblies along with their mutual connections in the grammatical (syntactic and semantic) network.


Adaptation of caprinised rinderpest virus to grow in Vero cells in vitro.
Caprinised rinderpest virus GTV strain (GTV) was adapted to grow in Vero cells (vGTV) by polyethylene glycol-mediated fusion of infected goat spleenocytes with non-infected Vero cells. The usual methods of infection of cell culture, i.e. virus adsorption or co-cultivation, were not successful. vGTV-induced cytopathic changes in Vero cells were similar to those reported for rinderpest virus. Virus titers increased with the passage number but the virulence for goats decreased. Immunoblot analysis did not reveal any difference between vGTV and vRBOK, the tissue culture rinderprest virus RBOK strain adapted to Vero cells.


Sigma-1 receptor ligands: potential in the treatment of neuropsychiatric disorders.
The sigma receptor was originally proposed to be a subtype of the opioid receptor. However, it is now clear that sigma receptors are unique non-opioid, non-phencyclidine brain proteins. Two types of sigma receptor exist, the sigma-1 receptor and the sigma-2 receptor. sigma-1 receptors have been cloned and their distribution, physiological functions and roles in signal transduction were recently characterised. Certain sex hormones in the brain (neurosteroids) are known to interact with sigma-1 receptors. sigma-1 receptors regulate glutamate NMDA receptor function and the release of neurotransmitters such as dopamine. They are thus proposed to be involved in learning and memory as well as in certain neuropsychiatric disorders. Selective sigma-1 receptor ligands have been suggested to represent a new class of therapeutic agents for neuropsychiatric disorders, although none have yet been introduced into therapeutic use. Early studies showed that psychotomimetic benzomorphans, as well as several antipsychotics, can bind to sigma-1 receptors. As a result of these findings, sigma-1 receptor ligands have been proposed as being of potential use in the treatment of schizophrenia. Nevertheless, the relationship of sigma-1 receptors to the underlying pathogenesis of schizophrenia is still unclear. sigma-1 receptor ligands have failed to improve acute psychotic symptoms of schizophrenia in clinical trials, but, interestingly, a few studies have shown an improvement in negative symptoms in schizophrenic patients. A number of preclinical studies have shown that selective agonists of sigma-1 receptors affect higher-ordered brain functions such as learning and memory, cognition and mood. These studies indicate that sigma-1 receptor agonists may exert therapeutic effects in depression and senile dementia. Indeed, the sigma-1 receptor agonist igmesine, has been shown to improve depression in a clinical trial. The most distinctive feature of the action of sigma-1 receptor ligands is their "modulatory" role. In behavioural studies of depression and memory, they exert beneficial effects only when brain functions are perturbed. Given the recently accumulated preclinical and clinical data, it is time to reconstruct the concept of sigma-1 receptors and the associated pathophysiological conditions that ligands of these receptors target. This would allow clinical trials to be performed more efficiently, and the results may confirm a long-speculated possibility that sigma-1 receptor ligands represent a new class of therapeutic agents for neuropsychiatric disorders.


Specific binding of 1,25-dihydroxycholecalciferol in human medullary thyroid carcinoma.
A specific 1,25-dihydroxycholecalciferol-binding protein has been detected in high-salt cytosols prepared from human medullary thyroid carcinomas. The binding protein had the same equilibrium dissociation constant (Kd = 0.17 +/- 0.05 nM; n = 4) and sedimentation coefficient on sucrose gradients (3.7S) as than seen in established vitamin D target tissues. This protein was not detected in normal thyroid cytosols, which may reflect the low proportion of C-cells within the gland.


Infants' perception of affordances of slopes under high- and low-friction conditions.
Three experiments investigated whether 14- and 15-month-old infants use information for both friction and slant for prospective control of locomotion down slopes. In Experiment 1, high- and low-friction conditions were interleaved on a range of shallow and steep slopes. In Experiment 2, friction conditions were blocked. In Experiment 3, the low-friction surface was visually distinct from the surrounding high-friction surface. In all three experiments, infants could walk down steeper slopes in the high-friction condition than they could in the low-friction condition. Infants detected affordances for walking down slopes in the high-friction condition, but in the low-friction condition, they attempted impossibly slippery slopes and fell repeatedly. In both friction conditions, when infants paused to explore slopes, they were less likely to attempt slopes beyond their ability. Exploration was elicited by visual information for slant (Experiments 1 and 2) or by a visually distinct surface that marked the change in friction (Experiment 3).


[Essex-Lopresti injury combined with forearm fracture].
The Essex-Lopresti injury is uncommon and often overlooked. A case is presented in which a 22-year old male sustained an injury that was a combination of an Essex-Lopresti injury and a fracture of the forearm. The primary treatment was osteosynthesis of the forearm. Ten weeks after injury the fractured radial head was removed because of pain and poor function of the elbow. The full extent of the injury was not recognized until sixteen weeks after injury. Final treatment consisted of excision of the ulnar head. The final results when seen at eight years follow-up were good.


MEMS-based tunable terahertz wire-laser over 330 GHz.
We demonstrate continuous tuning of a terahertz quantum cascade wire laser over a broad range of ∼330 GHz (∼8.6% of the 3.85 THz center frequency) with single-mode operation. Tuning is achieved with a narrow laser ridge (∼λ(o)/8) and a moveable MEMS (microelectromechanical systems) side object (plunger) to manipulate the transverse optical mode. The frictionless MEMS plunger enables continuous and reversible tuning. We demonstrate ∼30 GHz redshift tuning using a silicon plunger and ∼300 GHz blueshift tuning using a metal plunger. The blueshift tuning range is limited by the bandwidth of the gain medium rather than the tuning mechanism.


Patient satisfaction and results of vertical banded gastroplasty and gastric bypass.
Morbid obesity contributes to many health risks, including physical, emotional, and social problems. Various surgical treatments for morbid obesity have developed and have so far met with good results. This study compares vertical banded gastroplasty (VBG) with gastric bypass (GBP) and the patients' satisfaction with either procedure. Between April 1993 and July 1997, 63 bariatric surgical procedures were performed at Eisenhower Army Medical Center. Of those, complete follow-up was obtained for 29 patients. The parameters evaluated included age, preoperative and postoperative weights, body mass index (BMI), type of surgery, complications, and the patient's level of satisfaction. The study group consisted of 27 women and 2 men. The average preoperative weight was 135 kg, and the average preoperative BMI was 48.3 kg/m2. There were 17 VBGs and 12 GBPs performed. The average total weight loss was 45.1 kg. The average postoperative BMI was 33.2 kg/m2. There were no statistically significant differences in weight loss between VBG and GBP. Four of 17 patients had complications after VBG, and three of 12 patients had complications after GBP. After VBG, 94.1 % of patients were satisfied, and after GBP, 100% were satisfied. Twenty-seven of 28 patients stated that they would have the surgery again. There were no statistically significant differences in weight loss or complications after VBG or GBP. Patient satisfaction was high after both procedures. Therefore, bariatric surgery is important in the treatment of appropriately selected, morbidly obese patients.


Sanglifehrin A, a novel cyclophilin-binding immunosuppressant, inhibits IL-2-dependent T cell proliferation at the G1 phase of the cell cycle.
Sanglifehrin A (SFA) is a novel immunosuppressive natural product that binds to cyclophilin but is structurally distinct from cyclosporin A (CsA). We have investigated the cellular and molecular mechanisms of the action of SFA in T lymphocytes. We show that SFA inhibits T cell proliferation induced by IL-2 with an IC(50) of 200 nM. Distinct from CsA, which also binds to cyclophilin, SFA does not affect calcium-dependent IL-2 production, although SFA enhanced IL-2 gene transcription in the same cells. SFA blocks T cell proliferation induced by IL-2 in G(1) with no appreciable effect on IL-2 receptor expression in a manner similar to that of the immunosuppressant rapamycin. Unlike rapamycin, however, SFA has no effect on the phosphorylation or enzymatic activity of p70(s6k) kinase, distinguishing SFA from rapamycin in their mode of action. SFA inhibits hyperphosphorylation of Rb and the activity of cyclin E-cyclin-dependent kinase 2 on IL-2 signaling. These results suggest that SFA has a novel mode of action in comparison with CsA, FK506, and rapamycin, and that its use as a molecular probe may lead to the discovery of a novel target involved in T cell activation.


Postmarket surveillance of medical devices: current capabilities and future opportunities.
Recalls of cardiac implantable electrical devices (CIEDs) currently impact hundreds of thousands of patients worldwide. Premarket evaluation of CIEDs cannot be expected to eliminate all performance defects. Robust postmarket surveillance systems are needed to promote patient safety and reduce harm. Challenges impacting existing surveillance mechanisms include underreporting of defects, low rates of return of explanted CIEDs, lack of integration of surveillance into normal workflow, underutilization of existing resources including registries, a lack of capacity of aging resources, multiple proprietary platforms that lack interoperability, and the unmet need for common data variables as well as newer methods to generate, synthesize, analyze, and interpret evidence in order to respond rapidly to safety signals. Long-term solutions include establishing a unique device identification system; promoting expanded use of registries for surveillance and post-approval studies; developing additional methods to combine evidence from diverse data sources; creating tools and implementing strategies for universal automatic, triggered electronic event reporting; and refining methods to rapidly identify and interpret safety signals. Protection from litigation and creation of financial and other incentives by legislators, regulators, payers, accreditation organizations, and licensing boards can be expanded to increase participation in device surveillance by clinicians and health care facilities. Research to evaluate the comparative effectiveness of surveillance strategies is needed. Interim solutions to improve CIED surveillance while new initiatives are launched and the system strengthened are also presented.


Menstrual blood closely resembles the uterine immune micro-environment and is clearly distinct from peripheral blood.
Is menstrual blood a suitable source of endometrial derived lymphocytes? Mononuclear cells isolated from menstrual samples (menstrual blood mononuclear cells (MMC)) are clearly distinct from peripheral blood mononuclear cells (PBMC) and show a strong resemblance with biopsy-derived endometrial mononuclear cells. A critical event in the onset of pregnancy is the implantation of the embryo in the uterine wall. The immune cell composition in the endometrium at the time of implantation is considered pivotal for success. Despite advancing knowledge on the composition of the immune cell population in the uterus, the role of endometrial immune cells in reproductive disorders is still not fully resolved, mainly due to the fact that this type of research requires invasive techniques. Here, we collected menstrual fluid and validated this unique non-invasive technique to obtain and study the endometrium-derived immune cells which would be present around the time of implantation. Five healthy non-pregnant females with regular menstruation cycles and not using oral contraceptives collected their menstrual blood using a menstrual cup in five consecutive cycles. Sampling took place over the first 3 days of menses, with 12 h intervals. Peripheral blood samples, taken before and after each menstruation, were obtained for comparative analysis. MMC and PBMC samples were characterized for the different lymphocyte subsets by flow cytometry, with emphasis on NK cells and T cells. Next, the functional capacity of the MMC-derived NK cells was determined by measuring intracellular production of IFN-γ, granzyme B and perforin after culture in the presence of IL-2 and IL-15. In support of their endometrial origin, MMC samples contained the typical composition of mononuclear cells expected of endometrial tissue, were phenotypically similar to the reported phenotype for biopsy-derived endometrial cells, and were distinct from PBMC. Increased percentages of NK cells and decreased percentages of T cells were found in MMC when compared with PBMC from the same female. The MMC-derived NK cells were pre-dominantly CD56(bright)/CD16(-), in contrast to the primarily CD56(dim)/CD16(+) peripheral blood NK cells. MMC-derived NK cells expressed CD103, indicating their mucosal origin. In addition, the pattern of natural cytotoxicity receptor (NCR) expression in MMC-derived NK cells was comparable with that in endometrial biopsy-derived NK cells. Compared with PBMC, the NKp30 expression was decreased, while the percentage of NKp44 positive cells was increased in MMC samples. CXCR3 and CXCR4 were hardly expressed by MMC-derived NK cells, indicating that these cells are not of PBMC origin. NK cells from MMC samples were functional as shown by their capacity to produce IFN-γ, granzyme B and perforin, upon stimulation with IL-2 and IL-15. MMC-derived T cells revealed an increased expression of CD103, CD69 and CXCR4 compared with PBMC-derived T cells. Importantly, MMC collection using a menstrual cup proved highly reliable and reproducible between women and between cycles. Based on the parameters we studied, MMC appear similar to biopsy-derived endometrial mononuclear cells. However, sampling is not done at the exact same time in the menstrual cycle, and thus we cannot exclude some, as yet undetected, differences. Also, it should be considered that for some women, the use of the menstrual cup may be unpleasant. Menstrual blood may be a source of endometrial cells and may create new opportunities to study uterine immunological cells in fertility issues. No external funding was obtained for the present study. None of the authors have any conflict of interest to declare. NA.


Indian patients' attitudes towards chairside screening in a dental setting for medical conditions.
India has a high prevalence of cardiovascular disease (CVD), diabetes mellitus (DM), tuberculosis (TB), human immunodeficiency virus/acquired immune-deficiency syndrome (HIV/AIDS) and hepatitis B. United States-based studies indicate provider and patient support for medical screening in the dental setting. We assessed patient attitudes towards, and willingness to participate in, medical screening in the dental setting in India. A 5-point Likert scale survey (with scores ranging from 1=very important/willing to 5=very unimportant/unwilling) was given to a convenience sample of adult patients visiting five university-based dental clinics (clinic group) and one private-practice setting (private group). The Mann-Whitney U-test was used to compare mean response scores between patient groups. Logistic regression was used to assess factors associated with a favourable outcome. Both patient groups felt it important for dentists to identify increased risk for medical conditions (89.3% vs. 94.9%, respectively; P=0.02). The majority of patients were willing to have a dentist screen for the specified conditions: CVD (80.6% clinic and 84.5% private); DM (84.5% clinic and 77.5% private); TB (76.7% clinic and 73.2% private); hepatitis (73.3% clinic and 67.5% private); and HIV/AIDS (71.0% clinic and 70.5% private). The majority of patients were willing to participate in chairside screening that yielded immediate results (84.6% clinic and 86.1% private), discuss results immediately (85.8% clinic and 87.2% private) and pay 150 Indian rupees (55.9% clinic and 91.7% private). Younger patients (<40 years of age) were significantly less likely to respond favourably to: importance of medical screening in dental settings [adjusted odds ratio (OR)=0.63; 95% confidence interval (95% CI): 0.26-0.84] and be available for screening that yielded immediate results (adjusted OR=0.63; 95% CI: 0.40-0.99). Indian dental patients were in favour of chairside medical screening.


Pregnancy induced hypertension: evidence for increased cell membrane permeability to sodium.
In a cross sectional study of 137 women of childbearing age (16-40) the effects of normal pregnancy, hypertensive pregnancy, and oral contraceptives on red cell electrolyte content and sodium efflux rates were examined and the results compared with values in a control group of normotensive, non-pregnant women. Efflux rate constants were significantly increased in normotensive pregnancy and in women taking oral contraceptives. This was associated with a significant increase in sodium permeability in the contraceptive group. A much larger increase in sodium permeability and efflux rate constant was seen in the hypertensive group. The results permit a hypothesis that the hormonal changes induced by pregnancy and oral contraceptives increase membrane permeability to sodium and stimulate sodium efflux. The rise in blood pressure associated with use of oral contraceptives may have a similar aetiology to that occurring in pregnancy induced hypertension.


Genotype-renal function correlation in type 2 autosomal dominant polycystic kidney disease.
Autosomal dominant polycystic kidney disease (ADPKD) is a common Mendelian disorder that affects approximately 1 in 1000 live births. Mutations of two genes, PKD1 and PKD2, account for the disease in approximately 80 to 85% and 10 to 15% of the cases, respectively. Significant interfamilial and intrafamilial renal disease variability in ADPKD has been well documented. Locus heterogeneity is a major determinant for interfamilial disease variability (i.e., patients from PKD1-linked families have a significantly earlier onset of ESRD compared with patients from PKD2-linked families). More recently, two studies have suggested that allelic heterogeneity might influence renal disease severity. The current study examined the genotype-renal function correlation in 461 affected individuals from 71 ADPKD families with known PKD2 mutations. Fifty different mutations were identified in these families, spanning between exon 1 and 14 of PKD2. Most (94%) of these mutations were predicted to be inactivating. The renal outcomes of these patients, including the age of onset of end-stage renal disease (ESRD) and chronic renal failure (CRF; defined as creatinine clearance < or = 50 ml/min, calculated using the Cockroft and Gault formula), were analyzed. Of all the affected individuals clinically assessed, 117 (25.4%) had ESRD, 47 (10.2%) died without ESRD, 65 (14.0%) had CRF, and 232 (50.3%) had neither CRF nor ESRD at the last follow-up. Female patients, compared with male patients, had a later mean age of onset of ESRD (76.0 [95% CI, 73.8 to 78.1] versus 68.1 [95% CI, 66.0 to 70.2] yr) and CRF (72.5 [95% CI, 70.1 to 74.9] versus 63.7 [95% CI, 61.4 to 66.0] yr). Linear regression and renal survival analyses revealed that the location of PKD2 mutations did not influence the age of onset of ESRD. However, patients with splice site mutations appeared to have milder renal disease compared with patients with other mutation types (P < 0.04 by log rank test; adjusted for the gender effect). Considerable renal disease variability was also found among affected individuals with the same PKD2 mutations. This variability can confound the determination of allelic effects and supports the notion that additional genetic and/or environmental factors may modulate the renal disease severity in ADPKD.


Burnout among social workers in Iran: relations to individual characteristics and client violence.
Social workers are considered a professional group at high risk of burnout. Noticing the insufficient human resource management and understaffed social work centers, Iranian social workers are faced with a considerable level of physical and mental stress, which can lead to burnout. A national study on 390 social workers was conducted. Among social workers, 10.9% had experienced burnout and 17.4% are at risk of developing burnout. Social workers scored higher in burnout if they were dissatisfied with their income, had experienced violence, or had lower self-esteem. Findings are discussed with regard to Iranian context and recommendations for authorities of Iranian state welfare organizations are made.


G proteins, effectors and GAPs: structure and mechanism.
G proteins from a diverse family of regulatory GTPases which, in the GTP-bound state, bind to and activate downstream effectors. Structures of Ras homologs bound to effector domains have revealed mechanisms by which G proteins couple GTP binding to effector activation and achieve specificity. Complexes between structurally unrelated GTPase-activating proteins with complementary G proteins suggest common mechanisms by which GTP hydrolysis is stimulated via direct interactions with conformationally labile switch regions of the G protein.


Transcriptomic insights into antagonistic effects of gibberellin and abscisic acid on petal growth in Gerbera hybrida.
Petal growth is central to floral morphogenesis, but the underlying genetic basis of petal growth regulation is yet to be elucidated. In this study, we found that the basal region of the ray floret petals of Gerbera hybrida was the most sensitive to treatment with the phytohormones gibberellin (GA) and abscisic acid (ABA), which regulate cell expansion during petal growth in an antagonistic manner. To screen for differentially expressed genes (DEGs) and key regulators with potentially important roles in petal growth regulation by GA or/and ABA, the RNA-seq technique was employed. Differences in global transcription in petals were observed in response to GA and ABA and target genes antagonistically regulated by the two hormones were identified. Moreover, we also identified the pathways associated with the regulation of petal growth after application of either GA or ABA. Genes relating to the antagonistic GA and ABA regulation of petal growth showed distinct patterns, with genes encoding transcription factors (TFs) being active during the early stage (2 h) of treatment, while genes from the "apoptosis" and "cell wall organization" categories were expressed at later stages (12 h). In summary, we present the first study of global expression patterns of hormone-regulated transcripts in G. hybrida petals; this dataset will be instrumental in revealing the genetic networks that govern petal morphogenesis and provides a new theoretical basis and novel gene resources for ornamental plant breeding.


High-resolution structure of a mutagenic lesion in DNA.
The self-complementary dodecanucleotide d[CGC(m6G)AATTTGCG]2 (where m6G is O6-methylguanine), which contains two m6G.T base pairs, has been analyzed by x-ray diffraction methods and the structure has been refined to a residual error of R = 0.185 at 2.0-A resolution. The m6G.T mispair closely resembles a Watson-Crick base pair and there are very few structural differences between the m6G.T duplex and the native analogue. The similarity between the m6G.T base pair and a normal G.C base pair explains the failure of mismatch repair enzymes to recognize and remove this mutagenic lesion. A series of ultraviolet melting studies over a wide pH range on a related dodecamer indicate that the m6G.C mispair can exist in two conformations; one is a wobble pair and the other is a protonated Watson-Crick pair. The former, which predominates at physiological pH, will be removed by normal proofreading and repair enzymes, whereas the latter is likely to escape detection. Hence, the occasional occurrence of the protonated m6G.C base pair may explain why the presence of m6G in genomic DNA does not always give rise to a mutation.


Lunar crust: structure and composition.
Lunar seismic data from artificial impacts recorded at three Apollo seismometers are interpreted to determine the structure of the moon's interior to a depth of about 100 kilomneters. In the Fra Mauro region of Oceanus Procellarum, the moon has a layered crust 65 kilometers thick. The seismic velocities in the upper 25 kilometers are consistent with those in lunar basalts. Between 25 and 65 kilometers, the nearly constant velocity (6.8 kilometers per second) corresponds to velocities in gabbroic and anorthositic rocks. The apparent velocity is high (about 9 kilometers per second) in the lunar mantle immediately below the crust.


Optimal surgery for papillary thyroid carcinoma.
There has been a long, unresolved debate regarding the operation of choice for well differentiated carcinoma of the thyroid. We therefore analyzed the feasibility, scope, sample size, and length of follow-up required to determine the optimal operation for papillary thyroid carcinoma. A statistical approach was used to design a randomized prospective trial comparing the endpoints of complications, recurrence, and cause-specific mortality. A complication comparison trial is prohibitive owing to the large population required--approximately 12,000 randomized patients. A recurrence trial appears feasible based on sample size: approximately 360 to 800 patients with a 6 to 10-year follow-up. However, recurrence detection would be severely compromised in the lobectomy arm, and a unilateral lag-time bias would occur. A cause-specific mortality trial proves to be the least objectionable, although a large sample size (n = 3100) would be required. Such a trial is critically dependent on the ability to select an appropriate endpoint. A cause-specific mortality trial proves to be the most objective and statistically valid endpoint.


Clinical impact of ABL1 kinase domain mutations and IKZF1 deletion in adults under age 60 with Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL): molecular analysis of CALGB (Alliance) 10001 and 9665.
Recent studies have identified oncogenic lesions in Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL) and ABL1 kinase mutations that confer resistance to tyrosine kinase inhibitors. We sought to determine the prevalence and clinical impact of these lesions in patients on CALGB 10001, a previously reported Phase II study of imatinib, chemotherapy, and hematopoietic cell transplant in adult Ph + ALL. Of the 58 enrolled, 22 relapsed. By direct sequencing, an ABL1 kinase mutation known to induce imatinib resistance was present at relapse in 13 of 20. Using quantitative PCR assays, the mutations were detectable at diagnosis or early during treatment in most (62%) relapsed patients. Aberrations in IKZF1, CDKN2A/B, and PAX5 were assessed in 28 samples using SNP arrays and genomic DNA sequencing. Of these, 22 (79%) had IKZF1 deletion. The combination of IKZF1 deletion and p210 BCR-ABL1 (p < 0.0001), high white blood cell count (p = 0.021), and minimal residual disease (p = 0.013) were associated with worse disease-free survival.


The Role of Hypothalamic Pathology for Non-Motor Features of Huntington's Disease.
Huntington's disease (HD) is a fatal genetic neurodegenerative disorder. It has mainly been considered a movement disorder with cognitive symptoms and these features have been associated with pathology of the striatum and cerebral cortex. Importantly, individuals with the mutant huntingtin gene suffer from a spectrum of non-motor features often decades before the motor disorder manifests. These symptoms and signs include a range of psychiatric symptoms, sleep problems and metabolic changes with weight loss particularly in later stages. A higher body mass index at diagnosis is associated with slower disease progression. The common psychiatric symptom of apathy progresses with the disease. The fact that non-motor features are present early in the disease and that they show an association to disease progression suggest that unravelling the underlying neurobiological mechanisms may uncover novel targets for early disease intervention and better symptomatic treatment. The hypothalamus and the limbic system are important brain regions that regulate emotion, social cognition, sleep and metabolism. A number of studies using neuroimaging, postmortem human tissue and genetic manipulation in animal models of the disease has collectively shown that the hypothalamus and the limbic system are affected in HD. These findings include the loss of neuropeptide-expressing neurons such as orexin (hypocretin), oxytocin, vasopressin, somatostatin and VIP, and increased levels of SIRT1 in distinct nuclei of the hypothalamus. This review provides a summary of the results obtained so far and highlights the potential importance of these changes for the understanding of non-motor features in HD.


Chk1 modulates the interaction between myosin phosphatase targeting protein 1 (MYPT1) and protein phosphatase 1cβ (PP1cβ).
Polo-like kinase 1 (Plk1) is an instrumental kinase that modulates many aspects of the cell cycle. Previous investigations have indicated that Plk1 is a target of the DNA damage response, and Plk1 inhibition is dependent on ATM/ATR and Chk1. But the exact mechanism remains elusive. In a proteomic screen to identify Chk1-interacting proteins, we found that myosin phosphatase targeting protein 1 (MYPT1) was present in the immunocomplex. MYPT1 is phosphorylated by CDK1, thus recruiting protein phosphatase 1β (PP1cβ) to dephosphorylate and inactivate Plk1. Here we identified that Chk1 directly interacts with MYPT1 and preferentially phosphorylates MYPT1 at Ser20, which is essential for MYPT1-PP1cβ interaction and subsequent Plk1 dephosphorylation. Phosphorylation of Ser20 is abolished during mitotic damage when Chk1 is inhibited. The degradation of MYPT1 is also regulated by Chk1 phosphorylation. Our results thus unveil the underlying machinery that attenuates Plk1 activity during mitotic damage through Chk1-induced phosphorylation of MYPT1.


[A fluorescent method of determining aldosterone in urine].
The authors suggest a method of aldosterone determination in the urine. Aldosterone is extracted from the sample and purified with the aid of columnar and thin-layer chromatography on silicagel. Aldosterone is distinctly identified in ultraviolet light on the chromatogram by its green fluorescence developing after sprinkling the plate with phosphoric acid and its subsequent heating. Quantitative determination is carried out by the measurement of the intensity of aldosterone fluorescence in sulfuric acid.


A qualitative study of regional anaesthesia for vitreo-retinal surgery.
The aim of this research was to collect experiential knowledge about regional ocular anaesthesia - an integral component of most vitreo-retinal surgery. Anaesthesia for vitreo-retinal surgery has predominantly used general anaesthesia, because of the length and complexity of the surgical procedure. However, recent advances in surgical instrumentation and techniques have reduced surgical times; this decision has led to the adoption of regional ocular anaesthesia for vitreo-retinal day surgery. Although regional ocular anaesthesia has been studied from several perspectives, knowledge about patients' experience of the procedure is limited. An interpretive qualitative research methodology underpinned by Gadamer's philosophical hermeneutics. Eighteen participants were interviewed in-depth between July 2006-December 2007 following regional ocular anaesthesia. Interview data were thematically analysed by coding and grouping concepts. Four themes were identified: 'not knowing': the time prior to the experience of a regional eye block; 'experiencing': the experience of regional ocular anaesthesia; 'enduring': the capacity participants displayed to endure regional ocular anaesthesia with the hope that their vision would be restored; and 'knowing': when further surgery was required and past experiences were recalled. The experience of regional ocular anaesthesia had the capacity to invoke anxiety in the participants in this study. Many found the experience overwhelming and painful. What became clear was the participant's capacity to stoically 'endure' regional ocular anaesthesia, indicating the value people placed on visual function.


Photocatalytic CO(2) reduction using non-titanium metal oxides and sulfides.
Titanium dioxide (TiO2 ) is by far the most widely used photocatalyst both for the degradation of pollutants and in the field of renewable energies for the production of solar fuels. However, TiO2 has strong limitations in CO2 reduction, particularly under visible light irradiation. The flat-band potential of electrons in the conduction band of TiO2 is lower than that required for CO2 reduction and, therefore, it seems appropriate to develop and validate materials other than TiO2 . In addition, the photoresponse of TiO2 requires photons of wavelengths in the UV range shorter than 380 nm and strategies to implement a visible-light photoresponse on TiO2 by doping have not been completely satisfactory particularly because of problems in reproducibility and stability of the materials. For these reasons, we focus in this Review on semiconductors other than TiO2 that show photocatalytic activity in CO2 reduction. Attention has been paid to the irradiation conditions to put the productivity data into context. The role of co-catalyst and heterojunctions to increase the efficiency of charge separation is also discussed. Our aim is to describe the state of the art in the field of photocatalytic CO2 reduction using materials other than TiO2 , trying to trigger further research in this area.


Noninvasive Evaluation of CD20 Expression Using 64Cu-Labeled F(ab')2 Fragments of Obinutuzumab in Lymphoma.
CD20-overexpressed non-Hodgkin lymphoma typically indicates progressive malignancy. Obinutuzumab is a next-generation Food and Drug Administration-approved humanized monoclonal antibody that targets CD20. Previous studies with 89Zr-labeled obinutuzumab have successfully imaged CD20 in vivo. However, delayed tumor uptake and increased radioactive exposure caused by long blood circulation limit its clinical translation. This study aimed to develop 64Cu-labeled F(ab')2 fragments of obinutuzumab for imaging CD20 in lymphoma xenograft tumor models. Methods: F(ab')2 fragments were produced from obinutuzumab using an IgG-degrading enzyme of Streptococcus pyogenes (IdeS) enzyme and purified with protein A beads. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and high-performance liquid chromatography were performed to evaluate the products and their stability. F(ab')2 products were conjugated with p-SCN-Bn-NOTA (NOTA) for 64Cu radiolabeling. Western blotting was performed to screen the CD20 expression levels of lymphoma cells. Enzyme-linked immunosorbent assay, flow cytometry, and confocal imaging were used to test the binding affinity in vitro. Serial PET imaging and biodistribution studies in subcutaneous lymphoma-bearing mice were performed using 64Cu-NOTA-F(ab')2-obinutuzumab or 64Cu-NOTA-F(ab')2-IgG. Results: F(ab')2-obinutuzumab and F(ab')2-IgG produced by the IdeS digestion system were confirmed with sodium dodecyl sulfate polyacrylamide gel electrophoresis and high-performance liquid chromatography. The radiochemical purity of 64Cu-labeled F(ab')2 fragments was no less than 98%, and the specific activity was 56.3 ± 7.9 MBq/mg (n = 6). Among the 5 lymphoma cell lines, Ramos showed the strongest expression of CD20, and CLL-155 showed the lowest, as confirmed by enzyme-linked immunosorbent assay, flow cytometry, and confocal imaging. PET imaging revealed rapid and sustained tumor uptake of 64Cu-NOTA-F(ab')2-obinutuzumab in Ramos tumor-bearing mice. The peak tumor uptake (9.08 ± 1.67 percentage injected dose per gram of tissue [%ID/g]) in the Ramos model was significantly higher than that in the CCL-155 model (2.78 ± 0.62 %ID/g) or the 64Cu-NOTA-F(ab')2-IgG control (1.93 ± 0.26 %ID/g, n = 4, P < 0.001). The tumor-to-blood and tumor-to-muscle ratios were 7.3 ± 1.6 and 21.9 ± 9.0, respectively, at 48 h after injection in the 64Cu-NOTA-F(ab')2-obinutuzumab group. Of the measured off-target organs, the kidneys showed the highest uptake. Ex vivo immunofluorescent staining verified the differential CD20 expression in the Ramos and CCL-155 tumor models. Conclusion: This study demonstrated that 64Cu-NOTA-F(ab')2-obinutuzumab had a rapid and sustained tumor uptake in CD20-positive lymphoma with high contrast, which could enable noninvasive evaluation of CD20 levels in the clinic.


Phenotype characterization of human melanoma cells resistant to dabrafenib.
In the present study, the phenotype of melanoma cells resistant to dabrafenib (a B-RAF inhibitor) was investigated, to shed more light on melanoma resistance to B-RAF inhibition. Melanoma cells resistant to dabrafenib were generated using 3 different cell lines, A375, 397 and 624.38, all carrying B-RAFV600E, and they were characterized by cytofluorometric analysis, Ion Torrent technology, immunofluorescence and biochemistry. All dabrafenib-resistant cells showed, in addition to a re-activation of MAPK signaling, morphological changes compared to their sensitive counterparts, accompanied by an increase in CD90 (mesenchymal marker) expression and a decrease in E-cadherin (epithelial marker) expression, suggesting an epithelial-to-mesenchymal-like phenotypic transition. However, melanoma cells with TGF-β1-induced epithelial-to-mesenchymal transition (EMT) were more sensitive to dabrafenib treatment compared to the sensitivity noted in the non-TGF‑β1‑induced EMT melanoma cells, suggesting that TGF-β1-induced EMT was not associated with dabrafenib resistance. Although dabrafenib-resistant cells exhibited increased cell motility and E-cadherin/vimentin reorganization, as expected in EMT, all of them showed unvaried E-cadherin mRNA and unchanged Snail protein levels, while Twist1 protein expression was decreased with the exception of A375 dabrafenib-resistant melanoma cells, where it was unaffected. These findings suggest a distinct active EMT-like process adopted by melanoma cells under drug exposure. Furthermore, dabrafenib-resistant cells exhibited stem cell-like features, with Oct4 translocation from the cytoplasm to peri-nuclear sites and nuclei, and increased CD20 expression. In conclusion, our data, in addition to confirming that resistance to dabrafenib is dependent on re-activation of MAPK signaling, suggest that this resistance is linked to a distinct active EMT-like process as well as stem-cell features adopted by melanoma cells.


Lactose intolerance symptoms assessed by meta-analysis: a grain of truth that leads to exaggeration.
A meta-analysis was conducted to compare the lactose intolerance symptoms of lactose maldigesters after consuming lactose (as milk, lactose dissolved in water, milk products, or commercial product) with responses after a placebo under masked conditions. An English language MEDLINE search was conducted using the medical subject heading of "lactose intolerance" from 1966 to January 2002. From an initial 1,553 citations, 2 independent reviewers selected 21 studies based on study design (randomized, crossover, blind) and use of an amount of lactose likely to be found in a meal (7-25 g) and a placebo among subjects free of gastrointestinal problems and >4 years old. Mean severity of symptom responses were analyzed as standardized differences, and the presence or absence of a symptom was estimated as pooled incidence differences (ID). For severity of flatulence, the standardized difference was 0.18 (95% confidence interval [CI] -0.16 to +0.52). The CIs for abdominal bloating and pain, degree of diarrhea, frequency of bowel movements per day, and frequency of diarrhea per day also included 0. For abdominal bloating, the ID was 5.9 more people per 100 with symptoms after lactose than placebo (CI -0.07 to +0.19). This same nonsignificant relationship was found for abdominal pain. The ID for diarrhea or loose stools was 0.15 (CI 0.03 to 0.28). Although the incidence of diarrhea was significantly higher, the size of the effect was very small. The results indicate that lactose is not a major cause of symptoms for lactose maldigesters following usual intakes of dairy foods, that is, 1 cup.


3,4 methylenedioxymethamphetamine (ecstasy) impairs eight-arm radial maze performance and arm entry pattern in rats.
The recreational use of 3,4 methylenedioxymethamphetamine (ecstasy) in humans has been associated with memory impairment. The present study examined whether ecstasy impairs short- and long-term working memory and the pattern of arm entries in rats tested in the 8-arm radial maze with a 2-hr delay. After completing the training session, the rats were given a single dose of ecstasy (1, 2, or 3 mg/kg ip) 20 min before the test. The highest dose slightly affected short-term working memory. Under conditions of delay, there was a progressive deficit in long-term working memory, starting from 1 mg/kg. Under both test conditions, 2 and 3 mg/kg flattened the pattern of arm entry. None of the doses caused hyperlocomotion or stereotypy in the radial maze. These findings suggest that acute ecstasy mainly affects the long-term components of working memory and disrupts the pattern of arm entry in a way similar to serotonergic agents.


Variation in the use of diuretic therapy for infants with bronchopulmonary dysplasia.
To determine (1) between-hospital variation in diuretic use for infants with bronchopulmonary dysplasia (BPD), including hospital-specific treatment frequency, treatment duration, and percentage of infants receiving short (≤5 consecutive days) versus longer (>5 days) courses, and to determine (2) demographic and clinical variables associated with diuretic administration. A retrospective cohort study was conducted with the use of the Pediatric Health Information System to determine between-hospital variation in diuretic utilization patterns (primary outcome) and variables associated with diuretic use among <29-week-gestation infants with evolving BPD at age 28 days who were discharged between January 2007 and June 2011. During the 54-month study period, 1429 infants within 35 hospitals met the inclusion criteria for BPD at age 28 days, with 1222 (86%) receiving diuretic therapy for a median of 9 days (25th-75th percentile: 2-33 days). Short courses were administered to 1203 (83%) infants, and 570 (40%) infants received treatment for >5 consecutive days. Furosemide was the most widely prescribed diuretic (1218 infants; 85%), although chlorothiazide had the longest median duration of use (21 days; 25th-75th percentile: 8-46 days). The range of infants receiving a diuretic course of >5 days duration varied by hospital from 4% to 86%, with wide between-hospital variation even after adjustment for confounding variables. The frequency of diuretic administration to infants with BPD at US children's hospitals, as well as the specific diuretic regimen used, varies markedly by institution. Safety and effectiveness research of long-term diuretic therapy for BPD patients is needed to develop evidence-based recommendations.


300-kD subsynaptic protein copurifies with acetylcholine receptor-rich membranes and is concentrated at neuromuscular synapses.
Acetylcholine receptor-rich membranes from the electric organ of Torpedo californica are enriched in the four different subunits of the acetylcholine receptor and in two peripheral membrane proteins at 43 and 300 kD. We produced monoclonal antibodies against the 300-kD protein and have used these antibodies to determine the location of the protein, both in the electric organ and in skeletal muscle. Antibodies to the 300-kD protein were characterized by Western blots, binding assays to isolated membranes, and immunofluorescence on tissue. In Torpedo electric organ, antibodies to the 300-kD protein stain only the innervated face of the electrocytes. The 300-kD protein is on the intracellular surface of the postsynaptic membrane, since antibodies to the 300-kD protein bind more efficiently to saponin-permeabilized, right side out membranes than to intact membranes. Some antibodies against the Torpedo 300-kD protein cross-react with amphibian and mammalian neuromuscular synapses, and the cross-reacting protein is also highly concentrated on the intracellular surface of the post-synaptic membrane.


Fibula free flaps: the role of angiography in patients with abnormal results on preoperative color flow Doppler studies.
The reliability of normal color flow Doppler (CFD) study results in predicting the safety of fibula free flap harvest has been recognized. The significance of abnormal CFD study results when used for preoperative assessment of a potential fibula free flap donor site is less well defined. To determine if abnormal preoperative CFD study results should exclude fibula free flap harvest or if patients, in whom the fibula free flap is thought to be the best reconstructive option, should undergo further evaluation with angiography to better determine fibula free flap candidacy. A retrospective review identified 17 potential fibula free flap candidates (34 legs) evaluated by both a lower extremity CFD study and a lower extremity angiogram. The results of the CFD study were then compared with those of angiography. There were 16 legs with normal CFD study results and subsequent angiographic findings confirming the safety of each of these legs for fibula free flap harvest. There were 18 legs that demonstrated abnormal CFD study results. Angiography revealed anatomy that was considered to represent a high risk for fibula free flap harvest in 16 legs and considered safe in the other 2 legs. All 14 legs that had at least 1 vessel with a monophasic waveform or no flow on the CFD study revealed a high-risk angiogram result. Of the 4 legs with biphasic waveforms in all trifurcation vessels on the CFD study, 2 revealed angiogram results that showed that they were safe for flap harvest. Preoperative CFD studies that reveal a monophasic waveform or absence of flow accurately identify unsafe donor sites. Fibula free flap harvest in these cases can be excluded based on abnormal CFD study results alone, eliminating the need to perform angiography.


[Grief and depression in advanced age--the significance of losses of close relations in endogenous and neurotic-reactive depressions].
Largely "normal" grief reactions cannot always be easily separated from "pathological" grief reactions and depressive illness. Starting from methodological problems in grief research, 339 patients over 45 years who were treated for endogenous, neurotic or reactive depression and showed signs of pathological grief reaction after former or actual loss of a close-standing person were investigated. Marked sex differences were found. In the investigated patients, the symptom pattern of the pathological grief reactions contained mainly depressive developments, chronification of somatic complaints, phobic symptoms respectively anxious-neurotic developments, releasing of endogenous depressions, lack of grief, retarded grief reactions as well as drug abuse and suicidality. Particularly distressing factors which impede the "work of mourning" are shown in this paper. Finally, therapeutic aspects and fundamental problems and prospects of grief research are discussed.