Preparation of selectively metal-free and metal-substituted derivatives by reaction of Cu--Zn superoxide dismutase with diethyldithiocarbamate.
Incubation of Cu--Zn superoxide dismutase with diethyldithiocarbamate at increasing ligand/protein ratios and subsequent high-speed centrifugation led to proportional removal of copper from the protein, at variance with previous results [Misra (1979) J. Biol. Chem. 254, 11623--11628]. No zinc was lost, even at very high excesses of chelating agent. In this way a copper-free protein could be readily prepared, with avoidance of the critical pH condition and the dialysis step required in a previous method employing cyanide. The holoprotein was fully reconstituted from the copper-free protein by stoicheiometric re-addition of copper. From the mixture of metal-depleted forms originated by treatment with slight diethyldithiocarbamate excess, the protein containing copper only on one subunit, [Cu1--Zn2], could be isolated by preparative column electrophoresis. This species reproducibly showed 25% more specific activity (catalytic constant per copper) than that of the native or reconstituted [Cu2--Zn2] protein. This may result from long-range conformational effects between the active sites. By adding Co2+ ions to the vacant copper site of [Cu1--Zn2] a hybrid molecule containing Cu(II) on one subunit and Co(II) in the homologous site of the other subunit was prepared. Its activity, referred to copper, was identical with that of the native protein.


UV disinfection of indigenous aerobic spores: implications for UV reactor validation in unfiltered waters.
Conventional validation testing of UV reactors use cultured microorganisms spiked into test water flowing through a reactor. These tests are limited by the microbe titer it is possible to grow, thus limiting the size of the reactor it is possible to validate. The goal of this study was to examine the UV inactivation of indigenous aerobic spores naturally occurring in raw/unfiltered water supplies and to assess their use as an alternative indicator for validation testing of UV reactor performance, specifically for unfiltered water supplies planning large UV reactors. These spores were found in all raw waters tested in concentrations ranging between 20 and 12,000 CFU/100 mL and were very resistant to UV irradiation compared to a range of different microbes in the literature (i.e. adenovirus, MS-2 coliphage, and Cryptosporidium parvum). The inactivation of indigenous natural aerobic spores followed first-order kinetics with an inactivation coefficient ranging between 0.013 and 0.022 cm2/mJ with a high correlation coefficient. It was determined that naturally occurring aerobic spores, well characterized with respect to UV 253.7 nm inactivation, can be a useful tool when validating plant performance, and might also be used as a regular monitor of UV fluence and performance in a water treatment plant.


Glypican-1 targeted antibody-based therapy induces preclinical antitumor activity against esophageal squamous cell carcinoma.
Esophageal squamous cell carcinoma (ESCC) has a poor prognosis despite the development of multimodal therapy. Expression of glypican-1 (GPC1) has been reported to be elevated in a subset of patients with ESCC and associated with chemoresistance. This study aimed to determine the association of GPC1 with ESCC growth and potential usefulness of the GPC1 targeted therapy by monoclonal antibody (mAb) in ESCC. Expression of GPC1 was higher in ESCC tumor tissues than in adjacent non-tumoral tissues and normal tissues. Knockdown of GPC1 decreased growth of ESCC cells and induced apoptosis via inhibition of EGFR, AKT and p44/42-MAPK signaling pathways in vitro. Anti-GPC1 mAb strongly inhibited tumor growth via antibody-dependent cellular cytotoxicity dependent and independent manner in GPC1-positive ESCC xenograft models. Anti-GPC1 mAb also inhibited tumor growth of GPC1 positive ESCC patients derived tumor xenograft models. Furthermore, anti-GPC1 mAb showed a significant tumor growth inhibition with decreased angiogenesis compared with IgG treated controls in ESCC xenografted mice. Treatment with anti-GPC1 mAb was not toxic in mice. Anti-GPC1 mAb may have a potent anti-tumor effect and represent a novel treatment option for patients with GPC1-positive ESCC.


Impact of prior infection status on antibody response to the BNT162b2 mRNA COVID-19 vaccine in healthcare workers at a COVID-19 referral hospital in Milan, Italy.
In Italy, SARS-CoV-2 vaccination campaign prioritized healthcare workers (HCWs) to receive two doses of BNT162b2 vaccine, irrespective of a previous SARS-CoV-2 infection. In this real-life study, we compared the humoral response to BNT162b2 vaccine in HCWs with and without a previous SARS-CoV-2 infection. Of the 407 HCWs enrolled, 334 (82.1%) were SARS-CoV-2-naive and 73 (17.9%) SARS-CoV-2-experienced. Post-vaccine humoral response was detectable in more than 98% of HCWs. Overall, the median level of anti-S IgG in SARS-COV-2-experienced HCWs was twice as high as those of SARS-CoV-2-naive subjects (24641.0 AU/mL [IQR: 15273.0->40000.0] versus 13053.8 [IQR: 7303.3-20105.8]; p < .001), irrespective of the time elapsed from SARS-CoV-2 previous infection. In a subgroup of SARS-CoV-2-naive and -experienced subjects who received only one dose of the vaccine, the latter showed 32 times higher levels of anti-S IgG compared to the former. Although no serious adverse events have been reported, mild to moderate side effects occurred more frequently after the first dose in the SARS-CoV-2-experienced than in naive subjects (67% versus 42%, respectively; p < .001). Notably, post-vaccination anti-SARS-CoV-2 spike IgG levels ≥20,000 AU/mL were independently associated with the risk of fever ≥38°C (adjusted odds ratio [aOR] 5.122, 95% CI 2.368-11.080, p < .0001).Our study showed high responsiveness of BNT162b2 vaccine and a relationship between levels of antibody response and reactogenicity. It suggests that a single dose of mRNA vaccine might evoke effective protection in SARS-CoV-2-experienced subjects.


Rapid but incomplete degradation of residual visual representations over time.
While visual working memory has a short lifetime, residual representations can persist and disrupt currently maintained information. This phenomenon is known as proactive interference (PI), and the present study investigated whether the representations underpinning item-specific PI lose details over time. This would be expected if the memories underlying PI are susceptible to temporal processes such as decay, which is strongly disputed. In four experiments, a modified version of the recent probes task was used, requiring participants to determine whether a probe matched one of two recently presented targets. The probe sometimes matched an untested target from a previous trial, or varied in its resemblance to it, and the amount of time separating trials varied. Results revealed that PI was specific and highly disruptive at very short intervals, but its effect diminished over time. At longer intervals, a milder form of PI was present and produced by probes that were only similar to a recently encountered target. In summary, residual visual representations may remain accurate for a few seconds after encoding, before losing precise details and continuing to endure in an inexact state. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


OPIA: an open archive of plant images and related phenotypic traits.
High-throughput plant phenotype acquisition technologies have been extensively utilized in plant phenomics studies, leading to vast quantities of images and image-based phenotypic traits (i-traits) that are critically essential for accelerating germplasm screening, plant diseases identification and biotic & abiotic stress classification. Here, we present the Open Plant Image Archive (OPIA, https://ngdc.cncb.ac.cn/opia/), an open archive of plant images and i-traits derived from high-throughput phenotyping platforms. Currently, OPIA houses 56 datasets across 11 plants, comprising a total of 566 225 images with 2 417 186 labeled instances. Notably, it incorporates 56 i-traits of 93 rice and 105 wheat cultivars based on 18 644 individual RGB images, and these i-traits are further annotated based on the Plant Phenotype and Trait Ontology (PPTO) and cross-linked with GWAS Atlas. Additionally, each dataset in OPIA is assigned an evaluation score that takes account of image data volume, image resolution, and the number of labeled instances. More importantly, OPIA is equipped with useful tools for online image pre-processing and intelligent prediction. Collectively, OPIA provides open access to valuable datasets, pre-trained models, and phenotypic traits across diverse plants and thus bears great potential to play a crucial role in facilitating artificial intelligence-assisted breeding research.


The Vibrio cholerae O1 chromosomal integron.
Until the discovery of the Vibrio cholerae repeat (VCR), the gene capture and expression systems termed integrons had been typically associated with antibiotic-resistance gene cassettes with usually less than five genes in an array. A method is described for the cloning of the ends of large cassette arrays. Conserved restriction sites within VCRs facilitated the mapping by Southern hybridization and cloning of the 5' end of the VCR array, and using appropriate fragments it was possible to develop a physical map of the region of the V. cholerae chromosome. Sequence determination of the predicted beginning of this region revealed intI4, a member of the integron family of integrases. Comparison of these sequences from El Tor, Classical and serotype O134 V. cholerae strains identified the 3' end of the attI site, thereby defining the class 4 integron in one of the V. cholerae chromosomes, and providing the first evidence for integron-like site-specific recombination within V. cholerae. Conduction assays demonstrated IntI1-mediated recombination between VCRs. Restriction mapping places the sequences of intI4 and 26 VCR gene cassettes in arrays within a 120 kb region of the V. cholerae O1 strain 569B genome. This region contains an estimated 150 VCR gene cassettes, dwarfing previously described arrays. Southern analysis of genomic DNA from strains of Vibrio anguillarum, Vibrio mimicus and a number of V. cholerae serotypes revealed fragments that hybridized with VCR-specific probes but showed a high degree of restriction fragment length polymorphism. These data facilitate the identification of part of a new class 5 integron from V. mimicus.


[Factors associated with low knowledge on HIV/AIDS among men who have sex with men in Brazil].
This cross-sectional study analyzed social vulnerability, individual, and programmatic factors associated with low knowledge on HIV/AIDS among men who have sex with men (MSM). Respondent Driven Sampling (RDS) was used in 10 Brazilian cities. Knowledge scores on HIV/AIDS were estimated by Item Response Theory and categorized in three levels: high, medium, and low knowledge. Ordinal logistic regression was used for the analysis. Of the 3,746 MSM, 36.6%, 37.4%, and 26% showed high, medium, and low knowledge, respectively. The following were associated with low knowledge in the final model: schooling ≤ 8 years, non-white skin color, economic classes C and D-E, age < 25 years, having only one sex partner, and no previous HIV test. It is essential to improve knowledge on HIV/AIDS among young MSM with low socioeconomic status. Interventions focused on programmatic vulnerability factors can help reduce individual and social risk.


Multiquadrant digital analysis of shoulder capsular thickness.
Nonablative thermal capsular shrinkage has been developed in an attempt to address the plastic capsule deformation thought to cause increased rates of recurrent instability following arthroscopic stabilization procedures. Although the temperature required to optimize collagen shrinkage is known, a safe depth of thermal penetration, in various locations about the shoulder capsule, has not been defined. The purpose of this study was to measure shoulder capsule thickness by quadrant and circumferentially from the glenoid to the humerus so that thermal energy in shoulder procedures can be more precisely applied to limit possible injury to pericapsular structures. This is an anatomic study using a cadaveric shoulder specimens. Soft tissue was dissected from 8 fresh cadaveric shoulders to isolate intact glenohumeral joint capsules. The humeral insertion was released and the capsule was cut into 6 longitudinal quadrants around the glenoid. The capsule specimens were then flash frozen and stored at -80 degrees C. Quadrant tissue was cut into longitudinal sections 14 to 16 microm wide and stained with hematoxylin and eosin. The specimens were then digitized under a dissecting microscope and measured using computer imaging software at approximately 4-mm intervals. Two-way analysis of variance (ANOVA) was performed on the measurements of the intact capsule specimens 2.5 cm off the glenoid. Humeral insertion data were recorded separately. A total of 248 separate measurements were made throughout the capsule in 8 specimens. Capsular thickness increased from an average of 2.42 mm anteriorly to 2.80 mm in the inferior capsular pouch and again thinned to 2.22 mm posteriorly. Global shoulder capsule thickness ranged from 1.32 to 4.47 mm. When analyzed by position, from glenoid to humerus, a general thinning was noted with a mean thickness of 3. 03 mm at the glenoid to 2.17 mm at the humeral insertion. Two-way ANOVA showed a significant thickness variation along the specimen (P <.05), a nearly significant thickness variation with regard to quadrant (P <.03), and no significant interaction (P >.07) when applied to specimen measurements approximately 2.5 cm off the glenoid. The thickness of the shoulder capsule ranges from 1.32 to 4.47 mm, with a significant thinning laterally from the glenoid to the humerus. Further, capsule thickness ranges from 2.76 to 3.18 mm in the regions in closest proximity to the axillary nerve. These data may help determine the proper amount of thermal penetration necessary when performing shrinkage procedures and provide safety guidelines to limit the depth of thermal penetration to avoid possible injury to pericapsular structures.


Quantitative in vivo receptor binding. II. Autoradiographic imaging of muscarinic cholinergic receptors.
The in vivo distribution of [3H]scopolamine in rat brain following establishment of constant, saturating arterial tracer concentrations was examined with the use of quantitative autoradiography. The equilibrium drug distribution, studied 240 min after initiation of tracer infusion, was highly correlated with the regional density of muscarinic receptor sites determined in vitro in the same animals by autoradiographic analysis of [3H]quinuclidinyl benzilate binding. Brain regions of highest receptor density were generally correlated with known terminal fields of cholinergic neurons, and they demonstrated a protracted time course of in vivo labeling. An exception was noted in the basal pons, where a receptor population of high density without documented cholinergic innervation was rapidly labeled. It is suggested that synaptic muscarinic receptors are labeled slowly, as a consequence of either restricted tracer accessibility or competition between tracer and endogenous acetylcholine for available binding sites, and that the pontine receptors may be functionally distinct from those in other brain regions. The in vivo equilibrium binding technique used in the present study results in regional tissue radioligand concentrations directly proportional to receptor density and may, thus, provide a basis for receptor imaging in the human brain by means of positron emission tomography.


A firmer understanding of the effect of hypergravity on thyroid tissue: cholesterol and thyrotropin receptor.
Maintaining a good health requires the maintenance of a body homeostasis which largely depends on correct functioning of thyroid gland. The cells of the thyroid tissue are strongly sensitive to hypogravity, as already proven in mice after returning to the earth from long-term space missions. Here we studied whether hypergravity may be used to counteract the physiological deconditioning of long-duration spaceflight. We investigated the influence of hypergravity on key lipids and proteins involved in thyroid tissue function. We quantified cholesterol (CHO) and different species of sphingomyelin (SM) and ceramide, analysed thyrotropin (TSH) related molecules such as thyrotropin-receptor (TSHR), cAMP, Caveolin-1 and molecule signalling such as Signal transducer and activator of transcription-3 (STAT3). The hypergravity treatment resulted in the upregulation of the TSHR and Caveolin-1 and downregulation of STAT3 without changes of cAMP. TSHR lost its specific localization and spread throughout the cell membrane; TSH treatment facilitated the shedding of α subunit of TSHR and its releasing into the extracellular space. No specific variations were observed for each species of SM and ceramide. Importantly, the level of CHO was strongly reduced. In conclusion, hypergravity conditions induce change in CHO and TSHR of thyroid gland. The possibility that lipid rafts are strongly perturbed by hypergravity-induced CHO depletion by influencing TSH-TSHR interaction was discussed.


[Study of RNA and protein synthesis in mollusc epithelial tissues adapting to changes in the salinity of the medium].
RNA and protein synthesis in the epithelial tissues of White Sea snails Mytilus edulis was investigated autoradiographically with H-3-uridine and S-35-methionine during adaptation to various salinities (10 0/00, 14 0/00, 18 0/00, 35 0/00); normal salinity being 25 0/00. The incorporation of labelled precursors into the epithelial tissues of M. edulis was shown first to change abruptly, the return to the normal condition occurring after a prolonged adaptation. The reaction of different tissues shows specific features. The role of compensatory processes during the adaptation to changed salinities, is discussed.


Immunocytochemical evidence for translocation of protein kinase C in human megakaryoblastic leukemic cells: synergistic effects of Ca2+ and activators of protein kinase C on the plasma membrane association.
Immunological analysis using monoclonal antibodies against subspecies of protein kinase C revealed the predominant expression of the isozyme, type II, in human megakaryoblastic leukemic cells. We investigated the effects of phorbol diester 12-O-tetradecanoyl phorbol-13-acetate (TPA), the Ca2+ ionophore ionomycin and synthetic diacylglycerol 1-oleoyl-2-acetylglycerol (OAG) on the immunocytochemical localization of protein kinase C in these cells. Indirect immunofluorescence techniques revealed the enzyme to be located in a diffuse cytosolic pattern, in the intact cells. When the cells were exposed to 100 nM TPA, the immunofluorescent staining was translocated from the cytoplasm to the plasma membrane. The translocation was protracted and staining on the membrane decreased in parallel with the Ca2+, phospholipid-dependent protein kinase activity. Treatment of the cells with 500 nM ionomycin caused an apparent translocation comparable with that seen with TPA, however, this translocation was transient and most of the cytosolic staining was within 60 min. We also found that 30 micrograms/ml OAG did not have significant effects on distribution of the staining, but rather acted synergistically on the translocation with the suboptimal concentration of 100 nM ionomycin. A similar synergism was also observed with 10 nM TPA and 100 nM ionomycin. These results obtained in situ provide evidence that intracellular Ca2+ and diacylglycerol regulate membrane binding of the enzyme in vivo.


Energy expenditure and adiposity in Nigerian and African-American women.
Obesity is a prevalent condition in industrialized societies and is increasing around the world. We sought to assess the relative importance of resting energy expenditure (REE) and activity EE (AEE) in two populations with different rates of obesity. Women of African descent between 18 and 59 years of age were recruited from rural Nigeria and from metropolitan Chicago. Total EE (TEE) was measured using the doubly labeled water (DLW) technique and REE by indirect calorimetry; AEE was calculated as the difference between TEE and the sum of REE plus a factor for the thermic effect of food. In the analyses all EE parameters were adjusted for body size using a regression method. Comparisons were made between the groups and associations between EE and adiposity examined. A total of 149 Nigerian and 172 African-American women completed the protocol. All body size measurements were lower in the Nigerian women. Adjusted TEE and REE were higher in the Nigerian cohort but adjusted AEE did not differ significantly. Adjustment for parity, seasonality, and recent illness did not modify mean AEE or adiposity. In neither cohort was there a meaningful association between measures of AEE and adiposity. In these cohorts of women from very different environments, AEE did not differ significantly nor was it associated cross-sectionally with adiposity. If generalizable, these findings suggest that reduction in AEE may have less of a role in the development of obesity than anticipated. The possibility remains that variation in type and duration of activity plays a role not captured by total AEE.


[Experimental grounds for administration of probiotic drugs in conditions of hydroweightlessness].
We studied pharyngeal microflora in 22 healthy volunteers after 7-day dry immersion. For prophylactic pharyngeal dysbiosis, we used two probiotic drugs: oral -- "lactobacterin dry" and local --"lactobacterin immobilized on collagen". Administration of oral probiotic was accompanied with growth of pharyngeal opportunistic microflora preventing translocation of intestinal microflora. Local probiotic, on the contrary, decreased opportunistic microflora in the pharynx, but was associated with gastrointestinal dysbiosis. It is evident that combined use of topic and oral probiotic provides maximal effective prophylaxis of pharyngeal dysbiosis.


The adequate stimulus for avian short latency vestibular responses to linear translation.
Transient linear acceleration stimuli have been shown to elicit eighth nerve vestibular compound action potentials in birds and mammals. The present study was undertaken to better define the nature of the adequate stimulus for neurons generating the response in the chicken (Gallus domesticus). In particular, the study evaluated the question of whether the neurons studied are most sensitive to the maximum level of linear acceleration achieved or to the rate of change in acceleration (da/dt, or jerk). To do this, vestibular response thresholds were measured as a function of stimulus onset slope. Traditional computer signal averaging was used to record responses to pulsed linear acceleration stimuli. Stimulus onset slope was systematically varied. Acceleration thresholds decreased with increasing stimulus onset slope (decreasing stimulus rise time). When stimuli were expressed in units of jerk (g/ms), thresholds were virtually constant for all stimulus rise times. Moreover, stimuli having identical jerk magnitudes but widely varying peak acceleration levels produced virtually identical responses. Vestibular response thresholds, latencies and amplitudes appear to be determined strictly by stimulus jerk magnitudes. Stimulus attributes such as peak acceleration or rise time alone do not provide sufficient information to predict response parameter quantities. Indeed, the major response parameters were shown to be virtually independent of peak acceleration levels or rise time when these stimulus features were isolated and considered separately. It is concluded that the neurons generating short latency vestibular evoked potentials do so as "jerk encoders" in the chicken. Primary afferents classified as "irregular", and which traditionally fall into the broad category of "dynamic" or "phasic" neurons, would seem to be the most likely candidates for the neural generators of short latency vestibular compound action potentials.


A framework for direct locating and conformational sampling of protein structural motifs.
A specific treatment of recurrent structural motifs that represent the local bias information has been proven to be an important ingredient in de novo protein structure predication. Significant majority of methods for local structure are based on building blocks, which still suffer from its inherent discrete nature. Instead of using building blocks, this work presents a new protocol framework for local structural motifs prediction based on the direct locating along protein sequence and probabilistic sampling in a continuous (φ, ψ) space. The protein sequence was first scanned by an algorithm of sliding window with variable length of 7 to 19 residues, to match local segments to one of 82 motifs patterns in the fragment library. Identified segments were then labeled and modeled as the correlations of backbone torsion angles with mixture of bivariate cosine distributions in continuous (φ, ψ) space. 3D conformations of corresponding segments were finally sampled by using a backtrack algorithm to the hidden Markov model with single output of (φ, ψ). For local motifs in 50 proteins of testing set, about 62% of eight-residue segments located with high confidence value were predicted within 1.5 Å of their native structures by the method. Majority of local structural motifs were identified and sampled, which indicates the proposed protocol may at least serve as the foundation to obtain better protein tertiary structure prediction.


Discovery of a microbial transglutaminase enabling highly site-specific labeling of proteins.
Microbial transglutaminases (MTGs) catalyze the formation of Gln-Lys isopeptide bonds and are widely used for the cross-linking of proteins and peptides in food and biotechnological applications (e.g. to improve the texture of protein-rich foods or in generating antibody-drug conjugates). Currently used MTGs have low substrate specificity, impeding their biotechnological use as enzymes that do not cross-react with nontarget substrates (i.e. as bio-orthogonal labeling systems). Here, we report the discovery of an MTG from Kutzneria albida (KalbTG), which exhibited no cross-reactivity with known MTG substrates or commonly used target proteins, such as antibodies. KalbTG was produced in Escherichia coli as soluble and active enzyme in the presence of its natural inhibitor ammonium to prevent potentially toxic cross-linking activity. The crystal structure of KalbTG revealed a conserved core similar to other MTGs but very short surface loops, making it the smallest MTG characterized to date. Ultra-dense peptide array technology involving a pool of 1.4 million unique peptides identified specific recognition motifs for KalbTG in these peptides. We determined that the motifs YRYRQ and RYESK are the best Gln and Lys substrates of KalbTG, respectively. By first reacting a bifunctionalized peptide with the more specific KalbTG and in a second step with the less specific MTG from Streptomyces mobaraensis, a successful bio-orthogonal labeling system was demonstrated. Fusing the KalbTG recognition motif to an antibody allowed for site-specific and ratio-controlled labeling using low label excess. Its site specificity, favorable kinetics, ease of use, and cost-effective production render KalbTG an attractive tool for a broad range of applications, including production of therapeutic antibody-drug conjugates.


Breast-conserving surgery for invasive cancer: a principle based on segmental anatomy.
As the incidence of breast cancer increases in Japan, breast-conserving surgery becomes an important issue in the light of quality of life. We have demonstrated by 3-D reconstruction studies that ductal carcinoma in situ (DCIS) originates from the terminal duct-lobular unit (TDLU). Normal mammary epithelium anatomically located in the TDLU was shown to be biologically associated with cancerous change, particularly in specimens from patients who subsequently developed invasive carcinoma. Atypical ductal hyperplasia as well as DCIS expressed breast cancer associated antigen, providing further biological evidence that the atypical lesion at the TDLU are premalignant. Intraductal spread of carcinoma was defined as "DCIS was present clearly extending beyond the TDLU, or present prominently within the large ducts," and was classified into 3 grades according to the distribution of carcinoma in the duct-lobular system. We have developed a breast-conserving surgery consisting of quadrantectomy and regional lymph node dissection and immediate volume replacement using lateral tissue-flap (LTF). The quadrantectomy was employed on the basis of segmental anatomy of the duct-lobular system in which breast carcinoma originates. Fairly good cosmetic outcome as well as local control were obtained in the patients who underwent the immediate volume replacement using LTF. It must be emphasized that the quadrantectomy is a radical procedure in the sense that it aims at removal of all the carcinoma cells of the primary tumor.


Full characterization of modern transmission fibers for Raman amplified-based communication systems.
Telecommunication carriers have to estimate the Raman parameters of the fibers installed on their optical transport networks in order to facilitate the design of the next generation of high bit-rate Raman amplified-based transmission systems. This paper reports a very complete characterization of the most popular modern transmission fibers in terms of Raman efficiency, noise figure and double Rayleigh backscattering crosstalk. Our experiment is based on an averaged power analysis, applied to a counter-pumped long-haul distributed fiber Raman amplifier. We evaluate as well at 40 Gb/s for these different fiber types the double Rayleigh backscattering impact in terms of Q-factor penalty for various Raman gains and RZ modulation formats with different duty cycles.


Can molecular biomarker-based patient selection in Phase I trials accelerate anticancer drug development?
Anticancer drug development remains slow, costly and inefficient. One way of addressing this might be the use of predictive biomarkers to select patients for Phase I/II trials. Such biomarkers, which predict response to molecular-targeted agents, have the potential to enrich these trials with patients more likely to benefit. Doing so could maximize the efficiency of anticancer drug development by facilitating earlier clinical qualification of predictive biomarkers and generating valuable information on cancer biology. In this review, we suggest a new model of early clinical trial design, which incorporates patient selection through predictive molecular biomarkers for selected targeted agents.


CMV-specific CD8+ T-cell function is not impaired in chronic lymphocytic leukemia.
In chronic lymphocytic leukemia (CLL), CD8(+) T cells exhibit features of exhaustion and impaired functionality. Yet, reactivations of latent viruses such as cytomegalovirus (CMV) are uncommon in untreated CLL, suggesting that antiviral responses are uncompromised. We analyzed phenotypical and functional characteristics of CMV-specific CD8(+) T cells in CLL patients in comparison with age-matched healthy controls (HCs). Despite increased expression of the inhibitory receptors PD1, CD160, and CD244 on total CD8(+) T cells in CLL, expression levels of these markers were decreased on CMV-tetramer(+)CD8(+) T cells. Second, cytokine production upon stimulation with both phorbol 12-myristate 13-acetate/ionomycin and CMV-peptide-loaded antigen-presenting cells was intact in CMV-tetramer(+)CD8(+) T cells. Third, CMV-tetramer(+)CD8(+) T cells of CLL patients and HCs were equally effective in killing CMV-peptide-loaded target cells. Finally, quantitative imaging flow cytometry revealed that the proportion of CD8(+) T cells forming immunologic synapses with CMV-peptide-loaded B cells was intact. In conclusion, despite evidence for global T-cell dysfunction in CLL, we show here that CLL-derived CMV-specific CD8(+) T cells display lower expression of exhaustion markers and are functionally intact. These data indicate that the changes in the T-cell compartment in CLL may be more heterogeneous than presently assumed.


Influence of intake of skim milk from cows immunized with intestinal bacterial antigens on onset of renal disease in (NZB x NZW)F1 mice fed ad libitum or restricted in energy intake.
Autoimmune disease-prone (NZB x NZW)F1 (B/W) female mice are a model of human lupus erythematosus. Immune milk, obtained from cows immunized with various intestinal bacterial antigens, was given to B/W mice as a component of diets beginning at 8 wk of age. Diets were fed ad libitum or restricted to 60% of ad libitum energy intake. Controls were fed commercial skim milk. In B/W mice fed ad libitum, the titers of anti-single-stranded DNA antibodies were significantly lower in immune milk-fed mice at 4 and 6 mo of age. Onset of proteinuria was delayed and life span was significantly prolonged by immune milk feeding. Surface phenotypes of the T cells and levels of the responsiveness of lymphocytes to mitogens were not changed by immune milk feeding. The B/W mice restricted to 60% of ad libitum energy intake, which preserved immune responsiveness, had not developed proteinuria by 14 mo of age, irrespective of immune milk feeding or control milk feeding. However, at 10 mo of age, the level of plasma antibodies against intestinal bacteria was significantly higher in energy-restricted mice fed control milk than in those fed immune milk or in mice fed ad libitum.


Simvastatin over the counter.
In July 2004, simvastatin 10mg was reclassified in the UK from a Prescription-only medicine (PoM) to a Pharmacy (P) medicine and launched as Zocor Heart-Pro (Johnson & Johnson. MSD Consumer Pharmaceuticals). It is the first statin in the world to be sold without prescription. Zocor Heart-Pro is licensed to reduce the risk of a first major coronary heart disease (CHD) event in people likely to be at "moderate risk" of such problems. Here we review the rationale and process for reclassifying simvastatin 10mg as an over-the-counter product.


Laboratory growth of denitrifying water column microbial consortia from deep-sea shipwrecks in the northern Gulf of Mexico.
Background: Shipwrecks serve as a rich source for novel microbial populations that have largely remained undiscovered. Low temperatures, lack of sunlight, and the availability of substrates derived from the shipwreck's hull and cargo may provide an environment in which microbes can develop unique metabolic adaptations. Methods: To test our hypothesis that shipwrecks could influence the microbial population involved in denitrification when a consortium is grown in the laboratory, we collected samples proximate to two steel shipwrecks in the northern Gulf of Mexico. Then under laboratory conditions, we grew two independent denitrifying microbial consortia. Each consortium was grown by using the BART assay system and analyzed based on growth kinetics, ion chromatography and 16S amplicon sequencing. Results: Both denitrifying consortia were different from each other based on varied growth profiles, rates of nitrate utilization and 16S amplicon sequencing. Conclusions: Our observations conclude that the laboratory grown water column microbial consortia from deep-sea shipwrecks in the Gulf of Mexico are able to undergo aggressive denitrification.


Long-Term Follow-Up and Survivorship After Completing Systematic Surveillance in Stage I-III Colorectal Cancer: Who Is Still at Risk?
In patients with a high life expectancy at the time of surgery for colorectal cancer (CRC), the long-term outcome may be influenced by factors other than their cancer. We aimed to investigate the long-term outcome and cause of death beyond a 5-year surveillance programme. We evaluated the overall survival (OS) and cancer-specific survival (CSS) of a population-based cohort of stage I-III CRC patients <75 years old who completed a systematic surveillance programme. In total, 161 patients <75 years old, 111 (69 %) of whom were node negative (pN0), were included. The median follow-up time was 12.1 years. The OS was 54 % at 15 years and differed significantly between the pN0 and pN+ patients (65 vs. 30 %; P < 0.001); CSS (72 %) also differed between the pN0 and pN+ patients (85 vs. 44 %; P < 0.001). For the 5-year survivors (n = 119), 14 (12 %) died of CRC during additional long-term follow-up (7 each for pN0 and pN+), and 6 patients (5 %; all pN0) died of other cancers. Patients aged <65 years exhibited better long-term survival (81 %), but most of the deaths were due to CRC (10/12 deaths). Only two of the 14 cancer-related deaths involved microsatellite instable (MSI) CRC. Females exhibited better OS and CSS beyond 5 years of surveillance. The long-term survival beyond 5-year survivorship for stage I-III CRC is very good. Nonetheless, cancer-related deaths are encountered in one-third of patients and occur most frequently in patients who are <65 years old at disease onset-pointing to a still persistent risk several years after surgery.


Rate constants for hydrogen abstraction reactions by the hydroperoxyl radical from methanol, ethenol, acetaldehyde, toluene, and phenol.
An important step in the initial oxidation of hydrocarbons at low to intermediate temperatures is the abstraction of H by hydroperoxyl radical (HO(2)). In this study, we calculate energy profiles for the sequence: reactant + HO(2) → [complex of reactants] → transition state → [complex of products] → product + H(2)O(2) for methanol, ethenol (i.e., C(2)H(3)OH), acetaldehyde, toluene, and phenol. Rate constants are provided in the simple Arrhenius form. Reasonable agreement was obtained with the limited literature data available for acetaldehyde and toluene. Addition of HO(2) to the various distinct sites in phenol is investigated. Direct abstraction of the hydroxyl H was found to dominate over HO(2) addition to the ring. The results presented herein should be useful in modeling the lower temperature oxidation of the five compounds considered, especially at low temperature where the HO(2) is expected to exist at reactive levels.


Blood pressure pattern in urban Edos.
This study was part of a Hypertension Detection Programme carried out in Benin City in 1983. A sample of 1450 Edos aged 15-70 years (mean 31.8 years) had their blood pressures (BPs) recorded while sitting inside a bus; an automatic sphygmomanometer with cuff size 14 x 52 cm was used. Of these, 219 subjects (15.1%) had borderline hypertension while 168 (11.6%) were hypertensive (BP > or = 160/95 mmHg). There were more hypertensives among the 440 men aged 35-54 years compared with all other groups combined (P < 0.001). The highest age-specific prevalence (2.0%) was recorded in men aged 45-49 years. Mean BP rose with age until 39 years, after which there was a drop; thereafter the rise was noticed between 45 and 69 years. This study has highlighted a high prevalence of hypertension in the study population. Middle-aged men were found to be particularly vulnerable. There is need for mass enlightenment programmes about the hazards of hypertension especially in developing nations.


Stronger together: learning from an interdisciplinary dementia, arts and well-being network (DA&WN).
Background This paper reports on the learning from a 12-month interdisciplinary project (Dementia, Arts and Wellbeing Network- DA&WN) and its activities. These featured a series of four workshops on dance, visual art, theatre and music. The network was comprised of clinicians, academics, creative practitioners and people with lived experience of dementia and their carers. Methods The workshops were designed to draw out tacit knowledge about well-being in dementia through an action-based learning and research approach. This included, guided activities combined with reflective group discussions, visual documentation and baseline and follow-up questionnaires. Results Outcomes included new collaborations between group members, changes in creative practice for artists, and active and sustained involvement of people living with dementia and their carers in similar opportunities and participatory research. Conclusion This participatory and inclusive workshop model should be considered to develop and enhance interdisciplinary activities in dementia care.


Oncologic outcomes among Black and White men with grade group 4 or 5 (Gleason score 8-10) prostate cancer treated primarily by radical prostatectomy.
The aim of this study was to describe pathologic and short-term oncologic outcomes among Black and White men with grade group 4 or 5 prostate cancer managed primarily by radical prostatectomy. This was a multi-institutional, observational study (2005-2015) evaluating radical prostatectomy outcomes by self-identified race. Descriptive analysis was performed via nonparametric statistical testing to compare baseline clinicopathologic data. Univariable and multivariable time-to-event analyses were performed to assess biochemical recurrence (BCR), metastasis, cancer-specific mortality (CSM), and overall survival between Black and White men. In total, 1662 men were identified with grade group 4 or 5 prostate cancer initially managed by radical prostatectomy. Black men represented 11.3% of the cohort (n = 188). Black men were younger, demonstrated a longer time from diagnosis to surgery, and were at a lower clinical stage (all P < .05). Black men had lower rates of pT3/4 disease (49.5% vs 63.5%; P < .05) but higher rates of positive surgical margins (31.6% vs 26.5%; P = .14) on pathologic evaluation. There was no difference in BCR, CSM, or overall survival over a median follow-up of 40.7 months. Black men had a lower 5-year cumulative incidence of metastasis-free survival (93.6%; 95% confidence interval [CI], 86.5%-97.0%) in comparison with White men (85.8%; 95% CI, 83.1%-88.0%), which did not persist in an age-adjusted analysis. Black and White men with high-grade prostate cancer at diagnosis demonstrated similar oncologic outcomes when they were managed by primary radical prostatectomy. Our findings suggest that racial disparities in prostate cancer mortality are not related to differences in the efficacy of extirpative therapy.


Plasma metabolic profiling after cortical spreading depression in a transgenic mouse model of hemiplegic migraine by capillary electrophoresis--mass spectrometry.
Migraine is a common brain disorder characterized by recurrent attacks of severe headaches and other neurological symptoms. In one-third of patients headaches are accompanied by auras, which consist of transient visual and sensory disturbances, believed to be caused by cortical spreading depression (CSD). CSD is characterized by a wave of neuronal and glial depolarization with concomitant changes in metabolite concentrations in the brain and cerebrospinal fluid. It remains unknown whether CSD-induced brain metabolic changes can be captured outside the central nervous system, i.e., in peripheral fluids. This study investigated plasma metabolic changes in transgenic mice that harbor a gene mutation in voltage-gated CaV2.1 Ca(2+) channels previously identified in patients with familial hemiplegic migraine, a subtype of migraine with auras. The use of a mouse model allows investigation of molecular changes occurring shortly after CSD, which is notoriously difficult in patients. Capillary electrophoresis - mass spectrometry was used for the analysis of plasma samples to obtain, for the first time, a comprehensive view of molecular changes immediately after experimentally induced CSD. Multivariate data analysis showed a clear distinction between profiles of transgenic and wild-type animals after CSD. Two metabolites considered important for this discrimination were tentatively identified as being lysine and its by-product pipecolic acid with additional evidence provided by hydrophilic interaction chromatography combined with tandem mass spectrometry. The changed metabolites suggest a compensatory increase in GABAergic neurotransmission upon enhanced excitatory neurotransmission. These results show that CSD induces metabolic remodeling in transgenic migraine mice that can be captured and measured in plasma.


Torsional properties and microstructures of miniscrew implants.
Titanium miniscrew implants are popular in orthodontics, but there is little information about their torsional performance. Four brands of miniscrew implants (A-D) with 1.6-mm diameters were compared, with miniscrew A implants also having diameters of 1.2 to 2.0 mm. Nominal compositions of the implants were determined by x-ray fluorescence (n = 8). The miniscrews were loaded to failure in torsion, and the mean moment and twist angle were determined for each group (n = 8). Data were compared by ANOVA and the Tukey multiple range tests. Micro x-ray diffraction (n = 3) was used to identify phases in the implants, and the phases were also examined in etched cross-sections with a scanning electron microscope. Miniscrew A and C implants were pure titanium, whereas miniscrew B and D implants contained small amounts of vanadium, aluminum, iron, and manganese. Only alpha-titanium peaks were detected for all implants by micro x-ray diffraction, but beta titanium was observed in the microstructures of miniscrew B and D implants, which had significantly higher torsional moments at failure. Addition of small amounts of other elements to titanium yielded significantly improved torsional performance for miniscrew implants. Research to develop optimum compositions for mechanical properties and biocompatibility is needed.


Arrival of rabbit haemorrhagic disease virus 2 to northern Europe: Emergence and outbreaks in wild and domestic rabbits (Oryctolagus cuniculus) in Sweden.
Incursion of rabbit haemorrhagic disease virus (RHDV) into Sweden was documented in 1990 and it is now considered endemic in wild rabbit (Oryctolagus cuniculus) populations. Rabbit haemorrhagic disease virus 2 (RHDV2), a new, related lagovirus was first detected in France in 2010, and has spread rapidly throughout Europe and beyond. However, knowledge of RHDV2 in northern Europe is sporadic and incomplete, and in Sweden, routinely available diagnostic methods to detect rabbit haemorrhagic disease (RHD) do not distinguish between types of virus causing disease. Using RHDV2-specific RT-qPCR, sequencing of the VP60 gene and immunological virus typing of archived and prospective case material from the National Veterinary Institute's (SVA) wildlife disease surveillance programme and diagnostic pathology service, we describe the emergence of RHDV2 in Sweden in both wild and domestic rabbits. The earliest documented outbreak occurred on 22 May 2013, and from May 2013 to May 2016, 10 separate incidents of RHDV2 were documented from six different municipalities in the southern half of Sweden. Phylogenetic analysis of the VP60 gene shows clear clustering of Swedish isolates into three separate clusters within two different clades according to geographic location and time, suggesting viral evolution, multiple introduction events or both. Almost all cases of RHD examined by SVA from May 2013 to May 2016 were caused by RHDV2, suggesting that RHDV2 may be replacing RHDV as the predominant cause of RHD in Sweden.


Paradata: Measuring Engagement in Digital HIV Interventions for Sexual and Gender Minorities.
The goal of this review was to examine online engagement using paradata (i.e., intervention usage metrics) as part of the reporting of online behavioral HIV prevention and care interventions' findings. We underscore the importance of these data in examining intervention engagement and effectiveness. We focused on studies indexed in PubMed and published between April 1, 2017, and June 30, 2023, that reported the development and testing of online behavioral interventions for HIV prevention and/or care. Of the 689 extracted citations, 19 met the study criteria and provided engagement data - only six studies tested the association between engagement and intervention outcomes. Of these, four studies found a positive association between participants' engagement and improvements in HIV-related outcomes. Increasing attention is being paid to the collection and reporting of paradata within HIV online behavioral interventions. While the current evidence suggests a dose-response relationship due to user engagement on HIV outcomes, greater efforts to systematically collect, report, and analyze paradata are warranted.


The role of the Runx transcription factors in thymocyte differentiation and in homeostasis of naive T cells.
Members of the Runx family of transcriptional regulators are required for the appropriate expression of CD4 and CD8 at discrete stages of T cell development. The roles of these factors in other aspects of T cell development are unknown. We used a strategy to conditionally inactivate the genes encoding Runx1 or Runx3 at different stages of thymocyte development, demonstrating that Runx1 regulates the transitions of developing thymocytes from the CD4(-)CD8(-) double-negative stage to the CD4(+)CD8(+) double-positive (DP) stage and from the DP stage to the mature single-positive stage. Runx1 and Runx3 deficiencies caused marked reductions in mature thymocytes and T cells of the CD4(+) helper and CD8(+) cytotoxic T cell lineages, respectively. Runx1-deficient CD4(+) T cells had markedly reduced expression of the interleukin 7 receptor and exhibited shorter survival. In addition, inactivation of both Runx1 and Runx3 at the DP stages resulted in a severe block in development of CD8(+) mature thymocytes. These results indicate that Runx proteins have important roles at multiple stages of T cell development and in the homeostasis of mature T cells.


Effect of L-arginine on circulating endothelial progenitor cells and VEGF after moderate physical training in mice.
Alteration of levels and functional activities of circulating endothelial progenitor cells (EPCs) induced by risk factors for coronary heart disease (CHD) profoundly influence their role in the regeneration of tissue ischemia and angiogenesis. Among antioxidant nutrients in the prevention of CHD, L-arginine is particularly effective in enhancing the protection afforded by moderate physical exercise. Here, we aimed to evaluate the effects of L-arginine on EPC levels in C57BL/6J mice subjected to moderate physical exercise. Results showed that supplementation with L-arginine potentiates the effects of moderate physical exercise by increasing significantly EPCs (P<0.001) and VEGF serum levels (P<0.001). Our report highlights the beneficial effect of l-arginine in the modulation of EPC levels and VEGF secretion.


Safety profile of tinzaparin versus subcutaneous unfractionated heparin in elderly patients with impaired renal function treated for acute deep vein thrombosis: the Innohep® in Renal Insufficiency Study (IRIS).
Trials comparing the use of full dose unfractionated heparin (UFH) or low molecular weight heparins (LMWHs) in very elderly patients with impaired renal function are lacking. IRIS aimed to assess whether LMWH is at least as safe as UFH in this population. The study included renally impaired patients ≥70 years with acute symptomatic lower limb deep vein thrombosis (DVT). Patients were randomized to initial treatment with either tinzaparin 175 IU/kg once daily (n=269) or activated partial thromboplastin time-adjusted UFH twice daily (n=270). After acute management both groups received vitamin K antagonist to day 90. The trial was stopped prematurely due to a difference in mortality favoring the UFH group (11.5 vs. 6.3%; p=0.035). Rates of clinically relevant bleedings by day 90 were similar in the tinzaparin (11.9%) and UFH (11.9%) groups, as were rates of confirmed recurrent venous thromboembolism (VTE) (2.6 vs. 1.1%; p=0.34). As the mortality difference could not be explained by bleedings or recurrent VTE, a post-hoc analysis was performed. This identified six baseline characteristics significantly correlated with mortality, of which five were over-represented in the tinzaparin group. The IRIS study was a challenging study involving patients (mean age 83 years) usually excluded from clinical studies, but its early termination has left questions unanswered. The mortality difference observed with tinzaparin vs. UFH in elderly, renally-impaired patients with DVT cannot be explained on the basis of bleedings or recurrent VTE, and may reflect an imbalance of mortality risk factors at baseline.


Endoscope-assisted submandibular sialadenectomy: a review of outcomes, complications, and ethical concerns.
To review outcomes and complications of endoscope-assisted submandibular sialadenectomy (EASS) and to analyze this innovative technique with regard to ethical issues. We used a systematic review study design to identify clinical studies on EASS, published in English, French, German, and Thai. The last electronic search was conducted in September 2009. We checked the bibliographies of the identified articles, relevant local journals, and congress abstracts. Publications were further assessed and assigned their respective levels of evidence. We also investigated reporting on human subject protection, conflicts of interest, funding support, and commercial relationships. Five case series reporting a total of 28 patients met the inclusion criteria. There was no need of recourse to open surgery. All of the authors claimed satisfactory cosmetic results. Complications were uncommon. However, no controlled trial was available, and outcome measures varied between studies. Human subject protection and funding sources were mentioned in only 2 articles. Commercial relationships and conflicts of interest could not be identified. All of the reports favor outcomes of EASS. However, their level of evidence is low, and the superiority of this procedure over the conventional surgery remains unknown. The success of this procedure should not be overemphasized in information for consent and mislead surgeons to begin it without adequate training and elaborate environment. The lack of ethical documentation creates a high degree of suspicion of the studies.


Inequity in access to inpatient healthcare services for non-communicable diseases in India and the role of out-of-pocket payments.
Growing evidence suggests that non-communicable diseases (NCDs) result in considerable economic burden for individuals and households. With the poor facing a greater burden of NCDs than the rich in India, we undertook this study to analyse the horizontal equity in utilization and vertical equity in out-of-pocket expenditure for NCD care. We used data of 1 4 large Indian states from the National Sample Survey 60th round to compute hospitalization rates for NCDs. Mean per capita consumption expenditure (MPCE) was computed and used as a proxy measure for socioeconomic status. Out-of-pocket payment as a proportion of MPCE was estimated by wealth quintile (Q) to assess the vertical equity in payments. Concentration index (CoI) was computed to measure the extent of equity, and its 95% confidence interval was estimated to assess statistical significance. Overall, NCD hospitalizations in public facilities in India were used more by the poor (Col -0.041 ), while the rich used proportionately more services in the private sector (CoI 0.174). Out-of-pocket expenditure in public facilities was consistently lower than that in private facilities in urban and rural areas. The mean out-of-pocket expenditure for inpatient services for NCDs was found to be more among the rich in both public (Q5 ₹13 016, Q1 ₹4197) and private (Q5 ₹22 974, Q1 ₹8225) facilities. Public facilities are utilized more by poorer individuals. Strengthening the capacity of the public sector to deliver NCD care is required to meet equitable outcomes.


People thinking about thinking people. The role of the temporo-parietal junction in "theory of mind".
Humans powerfully and flexibly interpret the behaviour of other people based on an understanding of their minds: that is, we use a "theory of mind." In this study we distinguish theory of mind, which represents another person's mental states, from a representation of the simple presence of another person per se. The studies reported here establish for the first time that a region in the human temporo-parietal junction (here called the TPJ-M) is involved specifically in reasoning about the contents of another person's mind. First, the TPJ-M was doubly dissociated from the nearby extrastriate body area (EBA; Downing et al., 2001). Second, the TPJ-M does not respond to false representations in non-social control stories. Third, the BOLD response in the TPJ-M bilaterally was higher when subjects read stories about a character's mental states, compared with stories that described people in physical detail, which did not differ from stories about nonhuman objects. Thus, the role of the TPJ-M in understanding other people appears to be specific to reasoning about the content of mental states.


Dietary fibre and health: the role of hydrogen production.
The biological basis for the epidemiological association between high dietary fibre intakes and reduced disease incidence in Man is not fully understood. It is proposed that molecular hydrogen, produced in quite large amounts as a by-product of colonic fermentation of dietary fibre and unabsorbed carbohydrate may play an important role. Hydrogen in the tissues may act as a powerful reducing agent (antioxidant) of potentially damaging free radical species (e.g. singlet oxygen) and in so doing, along with other antioxidants would help in the control of these potentially damaging species and their known involvement in disease causation.


Improving sensitivity in nonlinear Raman microspectroscopy imaging and sensing.
Nonlinear Raman microspectroscopy based on a broadband coherent anti-Stokes Raman scattering is an emerging technique for noninvasive, chemically specific, microscopic analysis of tissues and large population of cells and particles. The sensitivity of this imaging is a critical aspect of a number of the proposed biomedical application. It is shown that the incident laser power is the major parameter controlling this sensitivity. By careful optimizing the laser system, the high-quality vibrational spectra acquisition at the multi-kHz rate becomes feasible.


Holographic generalized first-arriving light approach for resolving images viewed through a scattering medium.
We present a generalized holography-based approach with improved spatial resolution for extracting images, viewed through a scattering medium. The various angular directions are encoded either with different wavelengths or by capturing their corresponding images in different time slots. The various encoded images are recorded on a digital hologram with a computer. A digital reconstruction, which includes demodulation of the carrier beam and then a proper decoding algorithm, yields resolved images. The principle is demonstrated by recording image-plane digital holograms. Combining the suggested approach with the first-arriving light technique may further improve the results.


Stable and unstable fractures of the femoral shaft.
A retrospective review of 99 cases of stable and unstable femoral shaft fractures treated with Grosse-Kempf intramedullary fixation is presented. Despite minor complications and technical problems, the procedures resulted in union with acceptable alignment in 98% of cases. Details of the complications and technical problems are reviewed. There was no incidence of deep infection despite 16 open fractures. The two cases of nonunion were salvaged with revision internal fixation and bone grafting. Grosse-Kempf interlocking intramedullary fixation is a technically demanding procedure, but offers excellent results in the management of stable and unstable, open and closed fractures of the femoral shaft.


Yeast Rim11 kinase responds to glutathione-induced stress by regulating the transcription of phospholipid biosynthetic genes.
Glutathione (GSH), a tripeptide composed of glycine, cysteine, and glutamic acid, is an abundant thiol found in a wide variety of cells, ranging from bacterial to mammalian cells. Adequate levels of GSH are essential for maintaining iron homeostasis. The ratio of oxidized/reduced GSH is strictly regulated in each organelle to maintain the cellular redox potential. Cellular redox imbalances cause defects in physiological activities, which can lead to various diseases. Although there are many reports regarding the cellular response to GSH depletion, studies on stress response to high levels of GSH are limited. Here, we performed genome-scale screening in the yeast Saccharomyces cerevisiae and identified RIM11, BMH1, and WHI2 as multicopy suppressors of the growth defect caused by GSH stress. The deletion strains of each gene were sensitive to GSH. We found that Rim11, a kinase important in the regulation of meiosis, was activated via autophosphorylation upon GSH stress in a glucose-rich medium. Furthermore, RNA-seq revealed that transcription of phospholipid biosynthetic genes was downregulated under GSH stress, and introduction of multiple copies of RIM11 counteracted this effect. These results demonstrate that S. cerevisiae copes with GSH stress via multiple stress-responsive pathways, including a part of the adaptive pathway to glucose limitation.


Factors affecting patient compliance with compressive brace therapy for pectus carinatum.
The aim of this study was to identify factors affecting patient compliance with brace therapy for pectus carinatum. Eighty-six pectus carinatum patients who started brace therapy from August 2008 to November 2011 were included in this study. Patients were divided into two groups: patients who wore the brace for ≥6 months (compliance group) or patients who wore the brace for <6 months (non-compliance group). Factors affecting patient compliance were assessed at the last day of follow-up with a multiple-choice questionnaire. The questionnaire comprised seven items: pain at compression site, skin problems on compression area, confidence in brace treatment, shame, discomfort, initial result of bracing treatment and total number of factors affecting patient compliance. Eighty-six patients completed the survey, including seven (8.1%) female patients and 79 (91.9%) male patients, with a mean age of 12.0 years at the time of treatment (range, 3-20 years). The initial result of the compression period (P <0.001) and total number of factors affecting patient compliance (P <0.05) were significant predictors of patient compliance. An initial successful result of the compression period may increase patient compliance during treatment for pectus carinatum. Additional efforts to decrease pain, skin problems, shame and discomfort, and to give confidence may be beneficial in increasing compliance with bracing treatment.


Academic research: Foster defends DoD support in universities.
The United States has evolved a pattern of association between our universities and the Department of Defense. This association has led to major achievements in science and engineering, and has strengthened both national security and the national academic research base. Those who would make sweeping changes in this successfully evolving pattern should consider the whole case, all of the consequences for the country, from the many points of view. The issues must be tested in the larger context of all support to academic research, and the entire range of national groups and goals.


Conscientious refusal in healthcare: the Swedish solution.
The Swedish solution to the legal handling of professional conscientious refusal in healthcare is described. No legal right to conscientious refusal for any profession or class of professional tasks exists in Sweden, regardless of the religious or moral background of the objection. The background of this can be found in strong convictions about the importance of public service provision and related civic duties, and ideals about rule of law, equality and non-discrimination. Employee's requests to change work tasks are handled on a case-by-case basis within the frames of labour law, ensuring full voluntariness, and also employer's privilege regarding the organisation and direction of work, and duties of public institutions to provide services. Two complicating aspects of this solution related to the inclusion of 'alternative medical' service providers in a national health service, and professional insistence on conscientious refusal rights to accept legalised assisted dying are discussed. The latter is found to undermine the pragmatic reasons behind recent attempts by prolife groups to challenge the Swedish solution related to legal abortion in courts.


IncidencePrevalence: An R package to calculate population-level incidence rates and prevalence using the OMOP common data model.
Real-world data (RWD) offers a valuable resource for generating population-level disease epidemiology metrics. We aimed to develop a well-tested and user-friendly R package to compute incidence rates and prevalence in data mapped to the observational medical outcomes partnership (OMOP) common data model (CDM). We created IncidencePrevalence, an R package to support the analysis of population-level incidence rates and point- and period-prevalence in OMOP-formatted data. On top of unit testing, we assessed the face validity of the package. To do so, we calculated incidence rates of COVID-19 using RWD from Spain (SIDIAP) and the United Kingdom (CPRD Aurum), and replicated two previously published studies using data from the Netherlands (IPCI) and the United Kingdom (CPRD Gold). We compared the obtained results to those previously published, and measured execution times by running a benchmark analysis across databases. IncidencePrevalence achieved high agreement to previously published data in CPRD Gold and IPCI, and showed good performance across databases. For COVID-19, incidence calculated by the package was similar to public data after the first-wave of the pandemic. For data mapped to the OMOP CDM, the IncidencePrevalence R package can support descriptive epidemiological research. It enables reliable estimation of incidence and prevalence from large real-world data sets. It represents a simple, but extendable, analytical framework to generate estimates in a reproducible and timely manner.


Comparison of methods of performing the radioallergosorbent test: Phadebas, Fadal-Nalebuff and Hoffman protocols.
A series of control, IgE myeloma containing and atopic sera were tested by the radioallergosorbent test using identical lots of commercially available reagents following three different RAST protocols: Phadebas, Fadal-Nalebuff and that used in our own laboratories. In addition, a new RAST tracer was evaluated in the Phadebas and Fadal-Nalebuff methods. The Phadebas technique, using the standard 0.35 PRU cut-off, was the least sensitive at 68% but its sensitivity was raised to 84% using an 0.14 PRU cut-off. The specificity was 100% at 0.35 and 95% at 0.14 PRU. Our laboratory's technique gave a sensitivity of 84% and a specificity of 100%. The Fadal-Nalebuff method had the highest sensitivity at 92% but the lowest specificity at 92%. The Fadal-Nalebuff technique leads to a significant number of false positive scores, particularly in sera with IgE > 500 U/ml or with RAST discs that have significant non-specific binding. The Fadal-Nalebuff method can be used reliably if modified to include suitable negative controls for each antigen tested.


Measuring the organization of cardiac rhythms using the magnitude-squared coherence function.
The application of the magnitude-squared coherence (MSC) spectrum as a measure of the degree of organization of the cardiac electrical activity is explored. The MSC spectrum is a frequency-domain measure of the linear relationship between two signals. In the work described the two signals are two bipolar electrograms from either acutely placed catheter(s) or automatic implantable cardioverter/defibrillator electrodes. It is shown that the MSC is a dimensionless (no units), real-valued spectrum that is always in the range of zero to unity. The case of zero is found at frequencies where there is no linear relationship between the signals, and the case of unity implies a linear, noise-free relationship. The way the MSC spectrum is normalized makes it insensitive to gain or gain differences between the two signals. Example MSC spectra are presented and discussed. Striking differences in the spectra for fibrillatory and nonfibrillatory rhythms are seen.


A tale of the ciliate tail: investigation into the adaptive significance of this sub-cellular structure.
Ciliates can form an important link between the microbial loop and higher trophic levels primarily through consumption by copepods. This high predation pressure has resulted in a number of ciliate species developing rapid escape swimming behaviour. Several species of these escaping ciliates also possess a long contractile tail for which the functionality remains unresolved. We use high-speed video, specialized optics and novel fluid visualization tools to evaluate the role of this contractile appendage in two free-swimming ciliates, Pseudotontonia sp. and Tontonia sp., and compare the performance to escape swimming behaviour of a non-tailed species, Strobilidium sp. Here, we show that 'tailed' species respond to hydrodynamic disturbances with extremely short response latencies (less than or equal to 0.89 ms) by rapidly contracting the tail which carries the cell body 2-4 cell diameters within a few milliseconds. This provides an advantage over non-tailed species during the critical first 10-30 ms of an escape. Two small, short-lived vortex rings are created during contraction of the tail. The flow imposed by the ciliate jumping can be described as two well-separated impulsive Stokeslets and the overall flow attenuates spatially as r(-3). The high initial velocities and spatio-temporal arrangement of vortices created by tail contractions appear to provide a means for rapid escape as well as hydrodynamic 'camouflage' against fast striking, mechanoreceptive predators such as copepods.


Trisomy for the Down syndrome 'critical region' is necessary but not sufficient for brain phenotypes of trisomic mice.
Trisomic Ts65Dn mice show direct parallels with many phenotypes of Down syndrome (DS), including effects on the structure of cerebellum and hippocampus. A small segment of Hsa21 known as the 'DS critical region' (DSCR) has been held to contain a gene or genes sufficient to cause impairment in learning and memory tasks involving the hippocampus. To test this hypothesis, we developed Ts1Rhr and Ms1Rhr mouse models that are, respectively, trisomic and monosomic for this region. Here, we show that trisomy for the DSCR alone is not sufficient to produce the structural and functional features of hippocampal impairment that are seen in the Ts65Dn mouse and DS. However, when the critical region is returned to normal dosage in trisomic Ms1Rhr/Ts65Dn mice, performance in the Morris water maze is identical to euploid, demonstrating that this region is necessary for the phenotype. Thus, although the prediction of the critical region hypothesis was disproved, novel gene dosage effects were identified, which help to define how trisomy for this segment of the chromosome contributes to phenotypes of DS.


Flavin nucleotide metabolism in plants: monofunctional enzymes synthesize fad in plastids.
FAD synthetases (EC 220.127.116.11) catalyze biosynthesis of FAD from FMN and ATP. Monofunctional FAD synthetases are known to exist in mammals and yeast; bifunctional enzymes also catalyzing phosphorylation of riboflavin to FMN are known to exist in bacteria. Previously known eukaryotic enzymes with FAD synthetase activity have no sequence similarity to prokaryotic enzymes with riboflavin kinase and FAD synthetase activities. Proteins homologous to bacterial bifunctional FAD synthetases, yet shorter and lacking amino acid motifs at the C terminus, were found by bioinformatic analyses in vascular plant genomes, suggesting that plants contain a type of FAD synthetase previously known to exist only in prokaryotes. The Arabidopsis thaliana genome encodes two of such proteins. Both proteins, which we named AtRibF1 and AtRibF2, carry N-terminal extensions with characteristics of organellar targeting peptides. AtRibF1 and AtRibF2 cDNAs were cloned by reverse transcription-PCR. Only FAD synthetase activity was detected in the recombinant enzymes produced in Escherichia coli. FMN and ATP inhibited both enzymes. Kinetic parameters of AtRibF1 and AtRibF2 for the two substrates were similar. Confocal microscopy of protoplasts transformed with enhanced green fluorescence protein-fused proteins showed that AtRibF1 and AtRibF2 are targeted to plastids. In agreement with subcellular localization to plastids, Percoll-isolated chloroplasts from pea (Pisum sativum) synthesized FAD from imported riboflavin. Riboflavin kinase, FMN hydrolase, and FAD pyrophosphatase activities were detected in Percoll-isolated chloroplasts and mitochondria from pea. We propose from these new findings a model for subcellular distribution of enzymes that synthesize and hydrolyze flavin nucleotides in plants.


Environmental regulation, cost-benefit analysis, and the discounting of human lives.
The loss of human life resulting from environmental contaminants generally does not occur contemporaneously with the exposure to those contaminants. Some environmental problems produce harms with a latency period whereas others affect future generations. One of the most vexing questions raised by the cost-benefit analysis of environmental regulation is whether discounting, to reflect the passage of time between the exposure and the harm, is appropriate in these two scenarios. The valuations of human life used in regulatory analyses are from threats of instantaneous death in workplace settings. Discounting, to reflect that in the case of latent harms the years lost occur later in a person's lifetime, is appropriate in these circumstances. Upward adjustments of the value of life need to be undertaken, however, to account for the dread and involuntary nature of environmental carcinogens as well as for higher income levels of the victims. By not performing these adjustments, the regulatory process may be undervaluing lives by as much as a factor of six. In contrast, in the case of harms to future generations, discounting is ethically unjustified. It is simply a means of privileging the interests of the current generation. Discounting raises analytically distinct issues in the cases of latent harms and harms to future generations. In the case of latent harms, one needs to make intra-personal, intertemporal comparisons of utility, whereas in the case of harms to future generations one needs to define a metric against which to compare the utilities of individuals living in different generations. Thus, the appropriateness of discounting should be resolved differently in the two contexts.


Posterior Tibial Tendon Dysfunction: An Overview.
Adult acquired flatfoot deformity is a commonly seen condition with a large clinical spectrum. It ranges from asymptomatic subjects to severely disabled arthritic patients. Posterior tibialis tendon dysfunction is a common cause of adult acquired flatfoot deformity. This article systematically reviews the published literature from books and journals that were either originally written or later translated into the English language regarding the subject of posterior tibialis tendon dysfunction. Posterior tibialis tendon dysfunction is a primary soft tissue tendinopathy of the posterior tibialis that leads to altered foot biomechanics. Although the natural history of posterior tibialis tendon dysfunction is not fully known, it has mostly been agreed that it is a progressive disorder. While clinical examination is important in diagnosing adult acquired flat-feet; further investigation is often required to delineate the different aetiologies and stage of the disease. The literature describes many different management choices for the different stages of posterior tibialis tendon dysfunction. Because of the wide range of symptom and deformity severity, surgical reconstruction is based on a-la-carte. The consensus is that a plethora of reconstructive options needs to be available and the list of procedures should be tailored to tackle the different symptoms, especially when managing complex multi-planar reconstructions.


Passive protection of lambs against enteropathogenic Escherichia coli: role of antibodies in serum and colostrum of dams vaccinated with K99 antigen.
Lambs from suckling ewes vaccinated with the K99 antigen were resistant to challenge with K99-positive enteropathogenic Escherichia coli. Serum and colostrum from these ewes were compared with samples from control ewes to establish methods for monitoring vaccination and to determine the mechanism of protection. Vaccination stimulated production of K99 antibodies. These could be detected by an indirect haemagglutination test and a haemagglutination-inhibition test. Antiglobulin and gel-diffusion tests were less reliable. Experiments with brush-border cells from calf intestine showed that the antibodies were associated with anti-adhesive activity. The antibodies were predominantly IgG and did not neutralise the activity of heat-stable enterotoxin. It was concluded that neutralisation of the adhesive properties of the K99-positive E. coli by colostral antibodies significantly contributed to the resistance of the lambs from vaccinated ewes.


[The mutant gene wellhaarig disturbs the differentiation of hair follicle cells in the mouse].
First generation (G1) hairs in mice homozygous for the wellhaarig (we) gene are wavy and shorter than in normal mice; basal regions of the hairs are deformed. Follicles of G1 hairs in mid-dorsal region of 8-, 12- and 16-days old we/we mice were examined. Huxley cells of the inner root sheath (IRS) in apical region of hair follicles appeared to be hypertrophied. Cytoplasm of these cells was not stained by basic dyes and showed no birefringence. Cytoplasm of the IRS Henle cells was not stained by basic dyes either. These data indicate that keratinization of the IRS cells is disturbed in mutant homozygotes. The layer of outer root sheath in the we/we mice was thinner than in normal mice; this is probably due to hypertrophy of the IRS cells. The structure of differentiating cells of the hair shaft in normal and mutant mice was similar. The data obtained suggest that abnormal G1 hairs in we/we mice result from disturbance in IRS cells differentiation.


Ventral derotation spondylodesis. A review of 22 cases.
Twenty-two patients with major lumbar or thoracolumbar curves were treated with Zielke's modification of the Dwyer instrumentation, termed the "ventral derotation spondylodesis (VDS) system. In 16 patients, this was followed by planned second-stage posterior Harrington instrumentation and fusion. Six patients with adolescent idiopathic scoliosis were treated with VDS instrumentation and fusion alone. In neuromuscular and adult idiopathic scoliosis, a combined approach resulted in excellent curve correction and a high rate of successful fusion. In adolescent idiopathic scoliosis, VDS instrumentation alone resulted in excellent curve correction while permitting a shorter fusion length than conventional posterior Harrington instrumentation.


Resveratrol improves insulin sensitivity, reduces oxidative stress and activates the Akt pathway in type 2 diabetic patients.
Although resveratrol has widely been studied for its potential health benefits, little is known about its metabolic effects in humans. Our aims were to determine whether the polyphenol resveratrol improves insulin sensitivity in type 2 diabetic patients and to gain some insight into the mechanism of its action. After an initial general examination (including blood chemistry), nineteen patients enrolled in the 4-week-long double-blind study were randomly assigned into two groups: a resveratrol group receiving oral 2 × 5 mg resveratrol and a control group receiving placebo. Before and after the second and fourth weeks of the trial, insulin resistance/sensitivity, creatinine-normalised ortho-tyrosine level in urine samples (as a measure of oxidative stress), incretin levels and phosphorylated protein kinase B (pAkt):protein kinase B (Akt) ratio in platelets were assessed and statistically analysed. After the fourth week, resveratrol significantly decreased insulin resistance (homeostasis model of assessment for insulin resistance) and urinary ortho-tyrosine excretion, while it increased the pAkt:Akt ratio in platelets. On the other hand, it had no effect on parameters that relate to β-cell function (i.e. homeostasis model of assessment of β-cell function). The present study shows for the first time that resveratrol improves insulin sensitivity in humans, which might be due to a resveratrol-induced decrease in oxidative stress that leads to a more efficient insulin signalling via the Akt pathway.


Ion channel gating in cardiac ryanodine receptors from the arrhythmic RyR2-P2328S mouse.
Mutations in the cardiac ryanodine receptor Ca2+ release channel (RyR2) can cause deadly ventricular arrhythmias and atrial fibrillation (AF). The RyR2-P2328S mutation produces catecholaminergic polymorphic ventricular tachycardia (CPVT) and AF in hearts from homozygous RyR2P2328S/P2328S (denoted RyR2S/S) mice. We have now examined P2328S RyR2 channels from RyR2S/S hearts. The activity of wild-type (WT) and P2328S RyR2 channels was similar at a cytoplasmic [Ca2+] of 1 mM, but P2328S RyR2 was significantly more active than WT at a cytoplasmic [Ca2+] of 1 µM. This was associated with a >10-fold shift in the half maximal activation concentration (AC50) for Ca2+ activation, from ∼3.5 µM Ca2+ in WT RyR2 to ∼320 nM in P2328S channels and an unexpected >1000-fold shift in the half maximal inhibitory concentration (IC50) for inactivation from ∼50 mM in WT channels to ≤7 μM in P2328S channels, which is into systolic [Ca2+] levels. Unexpectedly, the shift in Ca2+ activation was not associated with changes in sub-conductance activity, S2806 or S2814 phosphorylation or the level of FKBP12 (also known as FKBP1A) bound to the channels. The changes in channel activity seen with the P2328S mutation correlate with altered Ca2+ homeostasis in myocytes from RyR2S/S mice and the CPVT and AF phenotypes.This article has an associated First Person interview with the first author of the paper.


Protective effects of a novel chimeric virus-like particle vaccine against virulent NDV and IBDV challenge.
Newcastle disease (ND) and infectious bursal disease (IBD) pose significant threats to the chicken industry, causing substantial economic losses. Currently, immunization through vaccination is the most effective strategy to prevent ND and IBD but currently used traditional vaccines, including inactivated or attenuated vaccines, face challenges in achieving a balance between immunogenicity and safety. To develop a green and efficient novel vaccine for ND and IBD, we developed a bivalent chimeric virus-like particle vaccine (ND-IBD cVLPs) displaying the ND virus (NDV) HN protein and the IBD virus (IBDV) VP2 protein based on the ND VLPs carrier platform and insect baculovirus expression system. This study aimed to evaluate the immunogenicity and protective efficacy of ND-IBD cVLPs in specific pathogen-free chickens. Chickens were immunized with 50 µg of purified ND-IBD cVLPs at 7 days old, boosted at 21 days old, and challenged at 42 days old. The results demonstrated that ND-IBD cVLPs stimulated highly effective hemagglutination inhibition antibody levels against NDV HN protein and enzyme-linked immunosorbent assay antibody levels against the IBDV VP2 protein. Furthermore, ND-IBD cVLPs provided complete protection against virulent NDV and IBDV challenges and mitigated pathological damage to the lung caused by NDV infection and the bursa of Fabricius caused by IBDV infection. These findings suggest that ND-IBD cVLPs hold promise as a safe and efficient novel vaccine candidate for the effective prevention of ND and IBD, extending the development of a foreign protein delivery platform of ND VLPs.


The metabolism of lipoprotein (a): an ever-evolving story.
Lipoprotein (a) [Lp(a)] is characterized by apolipoprotein (a) [apo(a)] covalently bound to apolipoprotein B 100. It was described in human plasma by Berg et al. in 1963 and the gene encoding apo(a) (LPA) was cloned in 1987 by Lawn and colleagues. Epidemiologic and genetic studies demonstrate that increases in Lp(a) plasma levels increase the risk of atherosclerotic cardiovascular disease. Novel Lp(a) lowering treatments highlight the need to understand the regulation of plasma levels of this atherogenic lipoprotein. Despite years of research, significant uncertainty remains about the assembly, secretion, and clearance of Lp(a). Specifically, there is ongoing controversy about where apo(a) and apoB-100 bind to form Lp(a); which apoB-100 lipoproteins bind to apo(a) to create Lp(a); whether binding of apo(a) is reversible, allowing apo(a) to bind to more than one apoB-100 lipoprotein during its lifespan in the circulation; and how Lp(a) or apo(a) leave the circulation. In this review, we highlight past and recent data from stable isotope studies of Lp(a) metabolism, highlighting the critical metabolic uncertainties that exist. We present kinetic models to describe results of published studies using stable isotopes and suggest what future studies are required to improve our understanding of Lp(a) metabolism.


Implementation of a quality system (ISO 9000 series) for placental blood banking.
Although placental blood has recently become a new source of hematopoietic progenitors for marrow replacement, limited attention has been given to systems suitable to ensure the short-term and long-term quality of placental blood units used for transplantation. In this article, we describe a quality system for placental blood banking developed in accord with ISO 9002 norms at Milano Cord Blood Bank. The quality system is the organizational structure, procedures, processes, and resources needed to implement quality management. ISO 9002 is a model for quality assurance in production, installation, and servicing, which includes a number of clauses providing guidance for the implementation of the quality system. The quality system was started by the bank medical director with step 1: the general quality plan, which included (a) the written description of mission, objectives, technical and organizational policies, and staff organization chart of the placental blood bank, (b) the definition and acquisition of adequate financial, human, and structural resources, (c) the appointment of a quality system head independent from the production laboratory and reporting directly to the medical director. Tasks of the quality system head were (a) to identify the placental blood banking process together with the placental blood bank personnel, (b) to implement a documentation plan finalized at the production and maintenance of (i) the quality manual, which provides a summary on how the bank operates with a quality system in compliance with the ISO 9002 clauses, (ii) the general procedures (or quality system procedures), which provide more detail on selected clauses, including at least those prescribed by the ISO 9002 standard, (iii) the operative procedures (or process procedures), which describe in detail the process of placental blood banking and how technical activities must be performed, (iv) the work instructions, which provide stepwise descriptions of individual activities, (v) records/forms for data collection and storage, (c) to identify quality indicators, (d) to start a regular internal audit, (e) to report audit results to the medical director for review. This was followed by step 2: the job descriptions, staff training, and qualification; step 3: the documentation plan; step 4: the internal audit plan; step 5: the launch of the quality system, and step 6: the assessment by an external team from an accredited third-party organization and final certification for compliance to ISO 9002. The quality system, which must be maintained and undergo external audit at regular intervals so that certification is confirmed, ensures the high probability that placental blood units provided to clinicians conform regularly to predefined levels of quality.


Clonal analysis of bilateral, recurrent, or systemically multifocal ocular adnexal lymphoma.
The purpose of this study is to determine the same or different clonality between bilateral or recurrent lesions, or between or among ocular adnexal lesions and systemically multifocal lesions in 10 consecutive patients with ocular adnexal lymphoma : 8 had extranodal marginal zone B-cell lymphoma of the mucosa-associated lymphoid tissue (MALT lymphoma) and 2 had mantle cell lymphoma, observed from 1995 to 2008 at Okayama University Hospital. A discrete DNA fragment was generated by polymerase chain reaction amplification of the immunoglobulin heavy chain gene from all samples except for two samples of bilateral orbital mantle cell lymphoma lesions and one sample of the scalp skin MALT lymphoma lesion. The size of DNA fragments were the same between bilateral orbital or conjunctival lesions of 5 patients, between original and recurrent conjunctival lesions of one patient, between the orbital lesion and the oral cavity lesion in one patient, and among bilateral orbital lesions, buccal, rectum, and stomach lesions of one patient. Sequencing of the DNA fragments showed the same sequence between bilateral or recurrent or multifocal lesions in 8 patients except for one : the bilateral orbital lesions, rectum, and stomach lesions shared the same sequence while the buccal lesion had one nucleotide difference compared to the sequence shared by the other 4 lesions. In conclusion, bilateral, recurrent, or systemically multifocal lesions of ocular adnexal lymphoma shared the clonality between or among the lesions.


Power, fresh water, and food from cold, deep sea water.
Many times more solar heat energy accumulates in the vast volume of warm tropic seas than that produced by all of our power plants. The looming energy crisis causes a renewal of interest in utilizing this stored solar heat to give, in addition to electric power, vast quantities of fresh water. Warm surface water, when evaporated, generates steam, to power a turbine, then fresh water when the steam is condensed by the cold water. A great increase in revenues over that from power and fresh water is shown by a substantial mariculture pilot plant. Deep sea water contains large quantities of nutrients. These feed algae which feed shellfish, ultimately shrimps and lobsters, in shallow ponds. Wastes grow seaweed of value; and combined revenues from desalination, power generation, and mariculture will give substantial profit.


Overexpression of apolipoprotein E4 increases kainic-acid-induced hippocampal neurodegeneration.
Apolipoprotein E (apoE) has an intricate biological function in modulating immune responses and apoE isoforms exhibit diverse effects on neurodegenerative and neuroinflammatory disorders. In the present study, we investigated the individual roles of apoE isoforms in the kainic acid (KA)-induced hippocampal neurodegeneration with focus on immune response and microglia functions. ApoE2, 3 and 4 transgenic mice as well as wild-type (WT) mice were treated with KA by intranasal route. ApoE4 overexpressing mice revealed several peculiarities as compared with other transgenic mice and WT mice, i.e. (1) they had more severe KA-induced seizures than apoE2 and 3 mice, (2) they exhibited neuron loss in hippocampus that was higher than in apoE2, 3 and WT mice, (3) KA administration resulted in higher counts of their head drops in the cross-area of elevated plus-maze, (4) they showed lower KA-induced rearing activity than apoE2 mice in the open-field test, (5) their KA-induced microglial expression of MHC-II and CD86 was elevated compared to apoE3 mice, (6) the KA-induced increase of microglial iNOS was higher than that in the other groups of mice, and (7) the TNF-α and IL-6 expression was decreased 7 days after KA application compared to untreated mice and mice treated 1 day with KA. However, the signaling pathway of NFκB or Akt seemed not to be involved in apoE-isoform dependent susceptibility to KA-induced neurotoxicity. In conclusion, over-expression of apoE4 deteriorated KA-induced hippocampal neurodegeneration in C57BL/6 mice, which might result from a higher up-regulation of microglia activation compared to apoE2 and 3 transgenic mice and WT mice.


Vitamin D3 inhibits expression and activities of matrix metalloproteinase-2 and -9 in human uterine fibroid cells.
Can biologically active vitamin D3 [1,25(OH)₂D3] regulate the expression and activity of matrix metalloproteinases (MMPs) in human uterine fibroid cells? 1,25(OH)₂D3 effectively reduced the expression and activities of MMP-2 and MMP-9 in cultured human uterine fibroid cells. Uterine fibroids (leiomyoma) express higher levels of MMP activity than adjacent normal myometrium, and this is associated with uterine fibroid pathogenesis. However, it is unknown whether 1,25(OH)₂D3 can regulate the expression and activities of MMPs in human uterine fibroid cells. Surgically removed fresh fibroid tissue was used to generate primary uterine fibroid cells. An immortalized human uterine fibroid cell line (HuLM) and/or primary human uterine fibroid cells isolated from fresh fibroid tissue were used to examine the expression of several MMPs, tissue inhibitors of metalloproteinases (TIMP) 1 and 2 and the activities of MMP-2 and MMP-9 after 1,25(OH)₂D3 treatment. Real-time PCR and western blots analyses were used to measure mRNA and protein expression of MMPs, respectively. Supernatant cell culture media were analyzed for MMP-2 and MMP-9 activities using a gelatin zymography assay. 1-1000 nM 1,25(OH)₂D3 significantly reduced mRNA levels of MMP-2 and MMP-9 in HuLM cells in a concentration-dependent manner (P < 0.5 to P < 0.001). The mRNA levels of MMP-1, MMP-3, MMP-13 and MMP-14 in HuLM cells were also reduced by 1,25(OH)₂D3. 1,25(OH)₂D3 significantly reduced MMP-2 and MMP-9 protein levels in a concentration-dependent manner in both HuLM and primary uterine fibroid cells (P < 0.05 to P < 0.001). Moreover, 1,25(OH)₂D3 increased the mRNA levels of vitamin D receptor (VDR) and TIMP-2 in a concentration-dependent manner in HuLM cells (P < 0.05 to P < 0.01). 1,25(OH)₂D3 also significantly increased protein levels of VDR and TIMP-2 in all cell types tested (P < 0.05 to P < 0.001). Gelatin zymography revealed that pro-MMP-2, active MMP-2 and pro-MMP-9 were down-regulated by 1,25(OH)₂D3 in a concentration-dependent manner; however, the active MMP-9 was undetectable. This study was performed using in vitro uterine fibroid cell cultures and the results were extrapolated to in vivo situation of uterine fibroids. Moreover, in this study the interaction of vitamin D3 with other regulators such as steroid hormone receptors was not explored. This study reveals an important biological function of 1,25(OH)₂D3 in the regulation of expression and activities of MMP-2 and MMP-9. Thus, 1,25(OH)₂D3 might be a potential effective, safe non-surgical treatment option for human uterine fibroids.


Green technologies for production of oils rich in n-3 polyunsaturated fatty acids from aquatic sources.
Fish and algae are the major sources of n-3 polyunsaturated fatty acids (n-3 PUFAs). Globally, there is a rapid increase in demand for n-3 PUFA-rich oils. Conventional oil production processes use high temperature and chemicals, compromising the oil quality and the environment. Hence, alternative green technologies have been investigated for producing oils from aquatic sources. While most of the studies have focused on the oil extraction and enrichment of n-3 PUFAs, less effort has been directed toward green refining of oils from fish and algae. Enzymatic processing and ultrasound-assisted extraction with environment-friendly solvents are the most promising green technologies for extracting fish oil, whereas pressurized extractions are suitable for extracting microalgae oil. Lipase-catalysed ethanolysis of fish and algae oil is a promising green technology for enriching n-3 PUFAs. Green refining technologies such as phospholipase- and membrane-assisted degumming deserve investigation for application in fish and algal oils. In the current review, we critically examined the currently existing research on technologies applied at each of the steps involved in the production of oils rich in n-3 PUFAs from fish and algae species. Special attention was placed on assessment of green technologies in comparison with conventional processing methods.


Exogenous choline augments transmission at an identified cholinergic synapse in terrestrial mollusk Limax maximus.
1. A variety of pharmacological tests indicate that the neuromuscular junction between the salivary burster neuron (SB) and the salivary duct muscle (SD) in the terrestrial mollusk Limax maximus is cholinergic. These include the effects of curare, atropine, and hexamethonium. 2. Exogenously applied choline can act both presynaptically and postsynaptically at the synapse between the SB and SD. a) When the SB-SD synapse is bathed in 30 microM choline, there is no measurable direct postsynaptic effect at the SB-SD synapse. After several hours of choline incubation, however, an increase is observed in the size of SB-elicited junction potentials recorded from the SD. b) When the synapse is bathed in 300 microM choline, a short-term decrease is seen in the amplitudes of junction potentials (JPs) recorded from the SD. This effect is interpreted as competition between choline and acetylcholine (ACh) for postsynaptic receptor sites on the SD, since the response of the denervated SD to ACh is diminished in the presence of 300 microM choline. c) After several hours incubation in 300 microM choline, the amplitudes of JPs elicited by the SB on the SD increase. 3. The long-term effect of exogenous choline is blocked by the choline-uptake inhibitor, hemicholinium 3. 4. We conclude that exogenous choline is taken up by the presynaptic terminals of the SB and converted to ACh. Transmitter output rises with growing transmitter stores, producing an increase in the size of SB-elicited JPs recorded from the SD.