Community-based educational intervention on necklace method as a natural family planning amongst reproductive age group women in India.
To assess the effectiveness of a community-based educational intervention on necklace method as a natural family planning amongst reproductive age group women. This approach helps women decide on their reproductive health choices and avoid ill health, impact and long-term consequences of unwanted pregnancy that lead to unsafe abortion. A total of 120 women were selected using non-probability purposive sampling technique. The knowledge and practice of participants were assessed using the structured knowledge questionnaire and practice checklist followed by a community-based educational intervention to participants, which covered aspects such as meaning, purposes and criteria; steps of the procedure; and advantages and disadvantages of the necklace method. The methods of teaching were lecture cum discussion and demonstration. The collected data were analysed using SPSS version 21. The levels of knowledge (t = 14.571, P = 0.023) and practice (t = 14.571, P = 0.026) significantly improved after administering the community-based educational intervention. Knowledge and practice positively correlated with each other (P < 0.05). Nurses play a vital role in educating women and creating awareness regarding modern and safe family planning methods. These methods are effective and essential to avoid unwanted pregnancy and thus greatly impact the health of women.


Soft modes of the dielectric response in the twist-bend nematic phase and identification of the transition to a nematic splay bend phase in the CBC7CB dimer.
The dielectric spectra of the twist-bend nematic phase (NTB) of (the bent-shaped) achiral liquid-crystal dimer 1''-,7''-bis(4-cyanobiphenyl-4'-yl)heptane (CB7CB) are studied for the determination of the different relaxation modes. Two molecular processes and one collective process were observed in the megahertz frequency range. Two molecular processes were assigned: one to the precessional rotation of the longitudinal components of the cyanobiphenyl groups and the second one to the spinning rotation of the transverse component of the CB7CB dimer. The peak, at a frequency of about 1 MHz, shows a peculiar temperature behavior at the NTB to N transition, reminiscent of the soft mode at the transition from the SmA to the SmC phase. This peak can be assigned to a collective fluctuation of the tilt angle of the coarse grained director N with respect to the pseudo-layer normal. This corresponds well with the electro-clinic effect observed as a response to an electric field in electro-optic experiments. The low frequency relaxation process, observed in the frequency range of 1 Hz-102 Hz can be identified as a Goldstone mode, related to long-scale fluctuation of the cone phase. The frequency drop of the mode on increasing the bias field is interpreted as unwinding of a helix and an indication of the formation of a field induced nematic splay bend phase (NSB). This finding is also confirmed by birefringence data in the presence of strong bias fields.


Activation of the skeletal muscle ryanodine receptor by suramin and suramin analogs.
Ca2+ release from skeletal muscle sarcoplasmic reticulum is activated by adenine nucleotides and suramin. Because suramin is known to interact with ATP-binding enzymes and ATP receptors (P2-purinergic receptors), the stimulation by suramin has been postulated to occur via the adenine nucleotide-binding site of the ryanodine receptor/Ca2+-release channel. We tested this hypothesis using suramin and the following suramin analogs: NF037, NF018, NF023, and NF007. The suramin analogs stimulate the binding of [3H]ryanodine binding to sarcoplasmic reticulum membranes with the following rank order of potency: suramin (EC50 = approximately 60 microM) > NF037 (EC50 = approximately 150 microM) > NF018 > NF023 > NF007. The suramin-induced stimulation occurs via a myoplasmic binding site on the ryanodine receptor as confirmed by binding experiments and single-channel recordings with the purified protein. This binding site is different than that for ATP, a conclusion that is supported by the following observations: (i) Suramin stimulates the association rate and inhibits the dissociation rate of [3H]ryanodine, whereas ATP analogs increase only the on-rate. (ii) In the presence of suramin but not of ATP analogs, [3H]ryanodine binding is resistant to the inhibitory effect of millimolar Mg2+ and Ca2+. (iii) ATP analogs and suramin have an additive effect on [3H]ryanodine binding. (iv) Affinity labeling of the purified ryanodine receptor with 2',3'-dialdehyde [alpha-32P]ATP or after in situ oxidation of [gamma-32P]ATP is not affected by suramin. Thus, our results show that suramin acts as a direct and potent stimulator of the ryanodine receptor but that this action is mediated via a binding site different from that for adenine nucleotides.


Comparative genome analysis indicates high evolutionary potential of pathogenicity genes in Colletotrichum tanaceti.
Colletotrichum tanaceti is an emerging foliar fungal pathogen of commercially grown pyrethrum (Tanacetum cinerariifolium). Despite being reported consistently from field surveys in Australia, the molecular basis of pathogenicity of C. tanaceti on pyrethrum is unknown. Herein, the genome of C. tanaceti (isolate BRIP57314) was assembled de novo and annotated using transcriptomic evidence. The inferred putative pathogenicity gene suite of C. tanaceti comprised a large array of genes encoding secreted effectors, proteases, CAZymes and secondary metabolites. Comparative analysis of its putative pathogenicity gene profiles with those of closely related species suggested that C. tanaceti likely has additional hosts to pyrethrum. The genome of C. tanaceti had a high repeat content and repetitive elements were located significantly closer to genes inferred to influence pathogenicity than other genes. These repeats are likely to have accelerated mutational and transposition rates in the genome, resulting in a rapid evolution of certain CAZyme families in this species. The C. tanaceti genome showed strong signals of Repeat Induced Point (RIP) mutation which likely caused its bipartite nature consisting of distinct gene-sparse, repeat and A-T rich regions. Pathogenicity genes within these RIP affected regions were likely to have a higher evolutionary rate than the rest of the genome. This "two-speed" genome phenomenon in certain Colletotrichum spp. was hypothesized to have caused the clustering of species based on the pathogenicity genes, to deviate from taxonomic relationships. The large repertoire of pathogenicity factors that potentially evolve rapidly due to the plasticity of the genome, indicated that C. tanaceti has a high evolutionary potential. Therefore, C. tanaceti poses a high-risk to the pyrethrum industry. Knowledge of the evolution and diversity of the putative pathogenicity genes will facilitate future research in disease management of C. tanaceti and other Colletotrichum spp.


Switching of molecular insertion in a cyclic molecule via photo- and thermal isomerization.
Two new cyclic ligands were synthesized: a ligand with two trans-azobenzene moieties and one bipyridine moiety, trans(2)-oAB-O13, and a ligand with two trans-azobenzene moieties and two bipyridine moieties, trans(2)-oAB-bpy. Both ligands underwent reversible trans-cis isomerization at the azobenzene moieties. The mole ratios of the trans(2) form:trans-cis form:cis(2) form, evaluated by (1)H NMR spectroscopy of the photostationary states prepared by 1 h illumination, were 0.13:0.27:0.60 (365 nm irradiation) and 0.41:0.47:0.12 (436 nm irradiation) for oAB-O13, and 0.18:0.12:0.70 (365 nm irradiation) and 0.36:0.43:0.21 (436 nm irradiation) for oAB-bpy. When trans(2)-oAB-O13 was mixed with Cu(I), both the bipyridine units and the polyether chains coordinated to the copper center. Addition of a noncyclic bipyridine ligand, trans(2)-oAB-2OH, afforded a bis(bipyridine)copper(I) complex, [Cu(trans(2)-oAB-O13)(trans(2)-oAB-2OH)]BF(4). The bis(bipyridine) ligand, trans(2)-oAB-bpy, formed a 1:1 complex with Cu(I), [Cu(trans(2)-oAB-bpy)]BF(4). [Cu(cis(2)-oAB-bpy)]BF(4) did not undergo the ligand substitution reaction with a noncyclic ligand with two azobenzene moieties and one bipyridine moiety, oAB, whereas its thermal isomerization in the presence of oAB caused the formation of [Cu(trans(2)-oAB-bpy)(trans(2)-oAB)]BF(4), indicating that the isomerization and ligand exchange reactions synchronized via a conformational change of the cyclic ligand.


On-Treatment Platelet Reactivity and Ischemic Outcomes in Patients With Diabetes Mellitus: Two-Year Results From ADAPT-DES.
Background Diabetes mellitus and high platelet reactivity (HPR) on clopidogrel are both associated with increased risk of ischemic events after percutaneous coronary intervention, but whether the HPR-associated risk of adverse ischemic events differs by diabetes mellitus status is unknown. Methods and Results ADAPT-DES (Assessment of Dual Antiplatelet Therapy With Drug-Eluting Stents) was a prospective, multicenter registry of patients treated with coronary drug-eluting stents. HPR was defined as P2Y12 reaction units >208 by the VerifyNow point-of-care assay. Cox multivariable analysis was used to assess whether HPR-associated risk of major adverse cardiac events (MACE; cardiac death, myocardial infarction, or stent thrombosis) varied for patients with insulin-treated diabetes mellitus (ITDM), non-ITDM, and no diabetes mellitus. Diabetes mellitus and HPR were included in an interaction analysis. Of 8582 patients enrolled, 2429 (28.3%) had diabetes mellitus, of whom 998 (41.1%) had ITDM. Mean P2Y12 reaction units were higher in patients with diabetes mellitus versus without diabetes mellitus, and HPR was more frequent in patients with diabetes mellitus. HPR was associated with consistently increased 2-year rates of MACE in patients with and without diabetes mellitus (Pinteraction=0.36). A significant interaction was present between HPR and non-insulin-treated diabetes mellitus versus ITDM for 2-year MACE (adjusted hazard ratio [HR] for non-ITDM, 2.28 [95% CI, 1.39-3.73] versus adjusted HR for ITDM, 1.02 [95% CI, 0.70-1.50]; Pinteraction=0.01). Conclusions HPR was more common in patients with diabetes mellitus and was associated with an increased risk of MACE in both patients with and without diabetes mellitus. In patients with diabetes mellitus, a more pronounced effect of HPR on MACE was present in lower-risk non-ITDM patients than in higher-risk patients with ITDM. Registration URL: https://clinicaltrials.gov/ct2/show/NCT00638794; Unique identifier: NCT00638794. ADAPT-DES (Assessment of Dual Antiplatelet Therapy With Drug-Eluting Stents).


Ticks of the province of Salamanca (Central/NW Spain). Prevalence and parasitization intensity in dogs and domestic ungulates.
By sporadic examination of 492 wild animals and regular weekly checks of dogs (179), cattle (288) and sheep and goats (141), 13 Ixodides were identified in the province of Salamanca. Some of these were only detected on the northern (I. ricinus) or southern (H. lusitanicum and B. annulatus) slopes of the mountains to the south of the province. Except for occasional introductions, H. lusitanicum and B. annulatus are not found on the Castillian plateau. In wild animals, 12 species were found of which only 4 are monotropic for some of these animals. In dogs, the most important species were R. turanicus and R. sanguineus. The latter shows only one generation per year and its nymphs are held to be the most important vector for the cases of boutonneuse fever registered in the provincial setting. In cattle, R. bursa is the most important tick, in terms of its prevalence in June/July (adults) and from November to January (immature forms). It also displays a much higher parasitization intensity than that of any of the other 7 species found on cattle. These remaining tick species (D. marginatus, H. marginatum, H. lusitanicum, I. ricinus, R. turanicus, Hae. punctata and B. annulatus) are abundant when R. bursa is absent; that is, in spring and at the end of summer, which thus are periods of richness in Ixodides but of low parasitism intensity. The ticks found on sheep and goats are the same as those detected on cattle, but in some of them prevalence and intensity vary as result of the kind of pasture frequented by the ovine and caprine hosts.


Fast, precise, and widely tunable frequency control of an optical parametric oscillator referenced to a frequency comb.
Optical frequency combs (OFCs) provide a convenient reference for the frequency stabilization of continuous-wave lasers. We demonstrate a frequency control method relying on tracking over a wide range and stabilizing the beat note between the laser and the OFC. The approach combines fast frequency ramps on a millisecond timescale in the entire mode-hop free tuning range of the laser and precise stabilization to single frequencies. We apply it to a commercially available optical parametric oscillator (OPO) and demonstrate tuning over more than 60 GHz with a ramping speed up to 3 GHz/ms. Frequency ramps spanning 15 GHz are performed in less than 10 ms, with the OPO instantly relocked to the OFC after the ramp at any desired frequency. The developed control hardware and software are able to stabilize the OPO to sub-MHz precision and to perform sequences of fast frequency ramps automatically.


[Capsular rupture of lymph node metastases as most significant prognostic factor in head and neck squamous cell carcinomas--Hamburg experiences].
TNM classification of squamous cell carcinomas of the head and neck contains only size and number of the lymph node metastases. The histological criterion of capsular rupture and its prognostic significance was assessed by an exact histological scheme of capsular rupture and extracapsular spread of the metastasis and was compared with N0 or lymph node metastases without capsular rupture respectively. Incidences of distant metastases, tumor recurrences, and survival of 194 consecutive patients with squamous cell carcinomas of the head and neck were investigated. Lymph node metastases were examined by the gradual histological scheme and patients were rated according their histological diagnosis. The classification N0/intranodal tumor growth/extranodal tumor growth resulted in a 5-year survival rate of 74%/62%/25%. According to the classification 67%/72%/37% of the patients were without local recurrent tumor and without distant metastases were 81%/79%/48% after 5 years. Capsular rupture seems to contain more decisive prognostic value as the criterion of only lymphatic metastatic disease. The histological scheme allows an exact assessment of the capsular rupture or the metastatic pattern of the lymph node. Because of its prognostic significance and individual therapeutic consequences the easily reproducible criterion of capsular rupture is a useful completion to current TNM classification.


Novel gigahertz frequency dielectric relaxations in chitosan films.
Molecular relaxations of chitosan films have been investigated in the wide frequency range of 0.1 to 3 × 10(9) Hz from -10 °C to 110 °C using dielectric spectroscopy. For the first time, two high-frequency relaxation processes (in the range 10(8) to 3 × 10(9) Hz) are reported in addition to the low frequency relaxations α and β. These two relaxation processes are related to the vibrations of OH and NH2/NH3(+), respectively. The high-frequency relaxations exhibit Arrhenius-type dependencies in the temperature range 10 °C to 54 °C with negative activation energy; this observation is traceable to hydrogen bonding reorientation. At temperatures above the glass transition temperature (54 °C), the activation energy changes from negative to positive values due to breaking of hydrogen bonding and water loss. Upon cooling in a sealed environment, the activation energies of two relaxation processes are nearly zero. FTIR and XRD analyses reveal associated structural changes upon heating and cooling. These two new high-frequency relaxation processes can be attributed to the interaction of bound water with OH and NH2/NH3(+), respectively. A plausible scenario for these high-frequency relaxations is discussed in light of impedance spectroscopy, TGA, FTIR and XRD measurements.


Sustainable nitrate-contaminated water treatment using multi cycle ion-exchange/bioregeneration of nitrate selective resin.
The sustainability of ion-exchange treatment processes using high capacity single use resins to remove nitrate from contaminated drinking water can be achieved by regenerating the exhausted resin and reusing it multiple times. In this study, multi cycle loading and bioregeneration of tributylamine strong base anion (SBA) exchange resin was studied. After each cycle of exhaustion, biological regeneration of the resin was performed using a salt-tolerant, nitrate-perchlorate-reducing culture for 48 h. The resin was enclosed in a membrane to avoid direct contact of the resin with the culture. The results show that the culture was capable of regenerating the resin and allowing the resin to be used in multiple cycles. The concentrations of nitrate in the samples reached a peak in first 0.5-1h after placing the resin in medium because of desorption of nitrate from resin with desorption rate of 0.099 ± 0.003 hr(-1). After this time, since microorganisms began to degrade the nitrate in the aqueous phase, the nitrate concentration was generally non-detectable after 10h. The average of calculated specific degradation rate of nitrate was -0.015 mg NO3(-)/mg VSS h. Applying 6 cycles of resin exhaustion/regeneration shows resin can be used for 4 cycles without a loss of capacity, after 6 cycles only 6% of the capacity was lost. This is the first published research to examine the direct regeneration of a resin enclosed in a membrane, to allow reuse without any disinfection or cleaning procedures.


Naive tinea corporis et cruris in an Immunocompetent adult caused by a geophile Nannizzia gypsea susceptible to Terbinafine-Rarity in the current scenario of Dermatophytosis in India.
Dermatophytosis caused by Nannizzia gypsea formerly Microsporum gypse um is rare in occurrence due to its geophilic adaptation and weak pathogenic potential in establishing infection in humans. The taxonomical status of N. gypsea has been controversial over the years and has now reached a concordance among mycologists. Innumerable reports of N. gypsea causing widespread infection in human immunodeficiency virus patients trails them as an important agent of consideration in an immunocompromised host. There have been sporadic reports of N. gypsea causing glabrous skin tinea and onychomycosis in healthy patients and the prevalence reports gravitate around 1-6.5 %. A variety of non-anthropophilic dermatophytes including novel species have now been implicated in causing dermatophytosis reflecting the era of crux changes in the epidemiology. We present a case of chronic dermatophytosis in a 22-year-old healthy Indian with a history of contact with a dog and soil and other factors favouring dermatophytosis. Conventional and molecular sequencing established the isolate as N. gypsea. Antifungal susceptibility test revealed a higher MIC of griseofulvin and lower MIC to azoles and terbinafine. The patient had complete clinical resolution following administration of oral terbinafine. Amidst the hyper-endemic-like scenario of tinea in India, this case report stands as a unique example of a patient infected with N. gypsea showing complete clinical resolution using terbinafine. Studies implicating N. gypsea in an immunocompetent host are rare and there is a need for more studies on geophilic dermatophytes causing tinea in the man for laying down effective preventive measures.


Thymoquinone ameliorates pulmonary vascular damage induced byEscherichia coli-derived lipopolysaccharide via cytokine downregulation in rats.
Our study investigated the ameliorative effects of thymoquinone (TQ) on the pulmonary blood vessels which were injured after intratracheal administration of Escherichia coli-derived lipopolysaccharide (LPS) in a rat model. Forty rats (150 ± 50 g) were randomly divided into four groups equally. The first group was intratracheally administered LPS (Escherichia coli O55:B5) at a dose 200 μg. The second group was co-administered intraperitoneal injection of TQ and LPS daily for one week. The third group was provided intraperitoneal injection of 1 mg of TQ. The fourth group was administered normal saline intratracheally at the rate of 200 μl. The results revealed that cytokine level of interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNFα) in serum was reduced in TQ-treated rats. Immunohistochemical study showed that expression of NF-kB was countered in the lung tissue by the application of TQ. In addition, the lesion score for various pathological aberrations were checked when rats were treated with TQ. From the results of the present study, it was concluded that TQ has an ameliorative effect on the pulmonary blood vascular damage via rearrangement of the cytokines in response to LPS injury in the rat model.


Elaiophylin Elicits Robust Anti-Tumor Responses via Apoptosis Induction and Attenuation of Proliferation, Migration, Invasion, and Angiogenesis in Pancreatic Cancer Cells.
Pancreatic cancer remains a formidable challenge in oncology due to its aggressive nature and limited treatment options. In this study, we investigate the potential therapeutic efficacy of elaiophylin, a novel compound, in targeting BxPC-3 and PANC-1 pancreatic cancer cells. We comprehensively explore elaiophylin's impact on apoptosis induction, proliferation inhibition, migration suppression, invasion attenuation, and angiogenesis inhibition, key processes contributing to cancer progression and metastasis. The results demonstrate that elaiophylin exerts potent pro-apoptotic effects, inducing a substantial increase in apoptotic cells. Additionally, elaiophylin significantly inhibits proliferation, migration, and invasion of BxPC-3 and PANC-1 cells. Furthermore, elaiophylin exhibits remarkable anti-angiogenic activity, effectively disrupting tube formation in HUVECs. Moreover, elaiophylin significantly inhibits the Wnt/β-Catenin signaling pathway. Our findings collectively demonstrate the multifaceted potential of elaiophylin as a promising therapeutic agent against pancreatic cancer via inhibition of the Wnt/β-Catenin signaling pathway. By targeting diverse cellular processes crucial for cancer progression, elaiophylin emerges as a prospective candidate for future targeted therapies. Further investigation of the in vivo efficacy of elaiophylin is warranted, potentially paving the way for novel and effective treatment approaches in pancreatic cancer management.


[Gastrointestinal hemorrhage--hemorrhage from the upper digestive system].
Upper gastrointestinal (GI) bleeding represents the commonest emergency managed by gastroenterologists utilizing substantial clinical and economic resources. Manifestations of GI bleeding depend uppon its localization, magnitude and co-morbidity. Although endoscopic haemostasis has significantly improved the outcome of patients with upper GI bleeding, in some cases patients continue to bleed or rebleed after initial control requiaring early elective surgery in order to decrease mortality. Despite recent advances in, both, endoscopic and surgical therapy, mortality rates have remained essentialy unchanged at 6-15%.


A protein-coding gene expression atlas from the brain of pregnant and non-pregnant goats.
Background: The brain is an extraordinarily complex organ with multiple anatomical structures involved in highly specialized functions related with behavior and physiological homeostasis. Our goal was to build an atlas of protein-coding gene expression in the goat brain by sequencing the transcriptomes of 12 brain regions in seven female Murciano-Granadina goats, from which three of them were 1-month pregnant. Results: Between 14,889 (cerebellar hemisphere) and 15,592 (pineal gland) protein-coding genes were expressed in goat brain regions, and most of them displayed ubiquitous or broad patterns of expression across tissues. Principal component analysis and hierarchical clustering based on the patterns of mRNA expression revealed that samples from certain brain regions tend to group according to their position in the anterior-posterior axis of the neural tube, i.e., hindbrain (pons and medulla oblongata), midbrain (rostral colliculus) and forebrain (frontal neocortex, olfactory bulb, hypothalamus, and hippocampus). Exceptions to this observation were cerebellum and glandular tissues (pineal gland and hypophysis), which showed highly divergent mRNA expression profiles. Differential expression analysis between pregnant and non-pregnant goats revealed moderate changes of mRNA expression in the frontal neocortex, hippocampus, adenohypophysis and pons, and very dramatic changes in the olfactory bulb. Many genes showing differential expression in this organ are related to olfactory function and behavior in humans. Conclusion: With the exception of cerebellum and glandular tissues, there is a relationship between the cellular origin of sampled regions along the anterior-posterior axis of the neural tube and their mRNA expression patterns in the goat adult brain. Gestation induces substantial changes in the mRNA expression of the olfactory bulb, a finding consistent with the key role of this anatomical structure on the development of maternal behavior.


Medicated early weaning to obtain pigs free from pathogens endemic in the herd of origin.
A field trial was conducted to assess the value of medicated early weaning for obtaining pigs free from some of the pathogens endemic in their herd of origin. The trial comprised 51 sows from a closed herd, which were farrowed in an isolated farrowing house in seven separate groups. The sows in each group were bred at the same time and induced to farrow on the same day. Their thriftiest piglets were weaned at five days of age and moved to an isolated early-weaning unit. At about six weeks of age they were moved to one of three isolated grow-out units where they were held to slaughter weight. Sows in five of the groups were dosed with high levels of tiamulin and trimethoprim-sulphonamide preparations from their entry into the farrowing house until their biggest piglets were weaned. Their piglets were dosed with similar drugs from birth until 10 days of age. The first and seventh groups of sows and their litters were not medicated. Tests were carried out on pigs aged five to 11 weeks, on slaughter pigs, and on pigs which died or were killed at different ages, for Mycoplasma hyopneumoniae, Bordetella bronchiseptica and colonic treponemes, which were readily detectable in the herd of origin. No evidence could be found of mycoplasma or bordetella. Colonic treponemes were recovered from some of the pigs at slaughter, but not from younger pigs. Thirty-seven boars and gilts from the medicated groups were introduced into 11 herds thought to be free of enzootic pneumonia and 13 were introduced into three herds which had enzootic pneumonia. No subsequent signs of enzootic pneumonia were noted in 10 of the enzootic pneumonia-free herds.


[The protein quality of commercial products of texturized soybean protein and meat mixtures].
The purpose of the present study was to evaluate the protein quality of eight commercial samples of texturized soybean protein (TSP) collected in the food markets of various Latin American countries. The study also included experiments designed to establish the limiting amino acids in those products, and studies to evaluate the process of texturization by extrusion cooking. Finally, experiments were also carried out to evaluate the effect of replacement of meat by TSP as well as the supplementary value of the latter to tortilla flour. The eight samples were characterized for their functional properties and protein quality. The results obtained indicate that all samples were different in density, water absorption, and nitrogen solubility. Likewise, all samples were different in their lysine, methionine and threonine content, with PER values ranging from 1.75 to 2.31. Protein quality standard value is 2.3. The amino acid supplementation studies showed a positive significant response to methionine addition, particularly in those products exhibiting low PER values. The addition of both lysine and threonine to the methionine-supplemented product, increased PER significantly. The data also demonstrated that replacement of more than 25% of meat protein by TSP protein decreased protein quality of the product, even when a TSP with an accepted protein quality value was used. This product was also found to supplement the quality of tortilla protein. Finally, it was established that the texturization process did not reduce the protein quality of soybean protein. Based on these results, it is concluded that the variability in the protein quality measured in the eight TSP samples was due to the initial quality of the soy flour texturized, which was already damaged before texturization. For this reason, it is recommended that Latin American countries establish TSP quality standards when this product is used as a protein extender for meat, and that appropriate systems be implemented for the quality control of such products.


The tryphostin AG17 induces apoptosis and inhibition of cdk2 activity in a lymphoma cell line that overexpresses bcl-2.
Tyrphostins are low molecular weight compounds that specifically inhibit protein tyrosine kinases. We studied the effects of tyrphostins on OCI-Ly8, a cell line derived from a patient with immunoblastic lymphoma that carries the t(14;18) translocation and overexpresses the B-cell lymphoma/leukemia-2 gene (bcl-2). To test the possibility that tyrphostins induce apoptosis in these cells, overcoming the protection rendered by bcl-2, we screened 16 tyrphostins representing different families at a concentration of 0.5-50 microM. We found that AG17 was the most potent in this regard. Cell cycle analysis demonstrated that AG17 induces arrest at the G1 phase followed by apoptosis with general reduction of the intracellular level of tyrosine-phosphorylated proteins. To further elucidate the mechanism of action of AG17, we investigated its effect on some of the key proteins that regulate the cell cycle. Bcl-2 and cdk2 protein levels were not altered with AG17, whereas cdk2 kinase activity, as well as p21 and p16 protein levels, were reduced markedly. These results suggest that the target of AG17 is inactivation of cdk2. Because lymphoma cells with the t(14;18) translocation and bcl-2 overexpression are resistant to chemotherapy, novel drugs selectively able to induce apoptosis in these cells could offer a new approach to the treatment of lymphoma patients.


SRC family kinases as potential therapeutic targets for malignancies and immunological disorders.
The Src family consists of eight non-receptor protein tyrosine kinases characterised by a common structure. Based on their amino acid sequence, Src family kinases are grouped into two subfamilies, which are also characterised by different tissue specificity. Src kinases are involved in signal transduction pathways triggered by a wide variety of surface receptors, including receptor tyrosine kinases, integrins, G-protein-coupled receptors and antigen receptors. Several pieces of evidence implicate Src family kinases in cancer development, as a consequence of changes in protein expression and/or kinase activity, and have prompted the design of potent specific inhibitors, the most common of which are adenine mimetics, as tools of relevant clinical interest for the treatment of both solid tumours and leukaemias. In addition, the finding that some Src kinases expressed in haematopoietic cells play pivotal roles in lymphocyte maturation and activation has fostered the development of safe and effective inhibitors selective for specific Src family members, which are currently being tested in clinical trials as immunosuppressants for the treatment of immunological disorders. Here we shall review the recent literature on the involvement of Src family kinases in human neoplasias and immunological disorders and the goals reached in the search for selective pharmacological inhibitors.


31P-NMR analysis of synaptic vesicles. Status of ATP and internal pH.
The status of ATP in cholinergic synaptic vesicles from electric organ of Torpedo marmorata has been studied by 31P-NMR. In isolated vesicles, ATP was found to be the only major phosphate-containing constituent. The chemical shifts and the linewidths of the P gamma and P beta resonances are different from those of uncompartmented ATP added to the suspension. The spectrum of intact electric tissue contains two sets of ATP resonances; one of them belongs to vesicular ATP as identified by its linewidths and chemical shifts. Both spectral characteristics of vesicular ATP, linewidths and chemical shifts, could be explained by an exchange mechanism, which also occurs in model solutions containing acetylcholine, ATP and Mg2+ at a pH around 5.5. We therefore conclude that the cholinergic synaptic vesicles store ATP together with acetylcholine and Mg2+ essentially in free solution at an acidic pH. Since the chemical shift of the P gamma resonance of ATP is largely determined by factors other than pH, the concept of its use as an indicator for the internal pH has to be modified in the case of these organelles.


Evaluating virtual endoscopy for clinical use.
Virtual endoscopy is a term used to describe computer simulated endoscopy procedures derived from high resolution images of patient anatomy. By simulating the endoscopic examination, the patient is spared the discomfort and possible complications of an actual examination. The physician also has more flexibility in a virtual endoscopic examination of 3D patient data in comparison to a real endoscopic examination. Virtual endoscopy removes the physical and physiologic constraints of real endoscopy and can create views that are not possible in an actual endoscopic examination. This may enhance the performance of actual endoscopic examinations. Virtual endoscopy may also be used to perform "numerical biopsies"; anatomic measurements such as size, distance, shape, and density. Virtual endoscopy allows the physician to comprehensively explore the patient anatomy using an intuitive and interactive interface. There are currently two technical approaches to performing virtual endoscopy: perspective volume rendering and surface rendering of polygonal models. Perspective volume rendering uses traditional volumetric rendering algorithms to create visualizations directly from the volumetric dataset. Polygonal models require a preprocessing step to convert the segmented volume information into a polygonal surface that may be displayed at real time frame rates. Both paradigms have inherent strengths and weaknesses. We illustrate and compare the methods on actual patient data, including simulated endoscopic examinations of the airways, colon and esophagus. Preliminary results in virtual endoscopy show promise and will continue to be an area of active research leading to useful clinical applications.


Management of hyponatremia and clinical use of vasopressin antagonists.
Hyponatremia, the most common electrolyte disorder in hospitalized patients, has been associated with high rate of mortality among both this population and nonhospitalized patients. This review describes briefly the classification and pathogenesis of hyponatremia, and, in greater detail, the management of hyponatremia with a particular emphasis on the clinical pharmacology of arginine vasopressin (AVP) antagonists. This review includes more in-depth discussion on the pharmacology of conivaptan, an AVP antagonist recently approved by the United States Food and Drug Administration.


A Study of the Bisphosphonic Derivatives from the Pudovik Reaction of Dialkyl α-Oxophosphonates and >P(O)H Reagents: X-ray Structure and Bioactivity.
New hydroxy-methylenebisphosphonic derivatives were prepared with different P-functions. The outcome of the reaction of α-oxophosphonates (YC(O)P(O)(OR)2) and dialkyl phosphites or diarylphosphine oxides depended on the Y substituent of the oxo-compound, the nature of the P-reagent and the amount of the diethylamine catalyst. Starting from dimethyl α-oxoethylphosphonate, in the presence of 5% of diethylamine, the corresponding Pudovik adduct was the single product. While using 40% of the catalyst, the rearranged species with the >P(O)-O-CH-P(O)< skeleton was the exclusive component. A similar reaction of α-oxobenzylphosphonate followed the rearrangement protocol. X-ray crystallography revealed not only the spatial structures of the three products, but also an intricate pattern evolving from the interplay of slight chemical differences, solvent inclusion and disorder as well as H-bridge patterns, which invite further investigation. In vitro activity of the compounds was assessed on different tumor cell cultures using end-point-type cell tetrazolium-based measurements. These structure-activity studies revealed a cytostatic effect for four rearranged derivatives containing aromatic units. One of them had a pronounced effect on MDA-MB 231 and Ebc-1 cells, showing IC50 = 37.8 and 25.9 µM, respectively.


The BR domain of PsrP interacts with extracellular DNA to promote bacterial aggregation; structural insights into pneumococcal biofilm formation.
The major human pathogen Streptococcus pneumoniae is a leading cause of disease and death worldwide. Pneumococcal biofilm formation within the nasopharynx leads to long-term colonization and persistence within the host. We have previously demonstrated that the capsular surface-associated pneumococcal serine rich repeat protein (PsrP), key factor for biofilm formation, binds to keratin-10 (KRT10) through its microbial surface component recognizing adhesive matrix molecule (MSCRAMM)-related globular binding region domain (BR187-385). Here, we show that BR187-385 also binds to DNA, as demonstrated by electrophoretic mobility shift assays and size exclusion chromatography. Further, heterologous expression of BR187-378 or the longer BR120-378 construct on the surface of a Gram-positive model host bacterium resulted in the formation of cellular aggregates that was significantly enhanced in the presence of DNA. Crystal structure analyses revealed the formation of BR187-385 homo-dimers via an intermolecular β-sheet, resulting in a positively charged concave surface, shaped to accommodate the acidic helical DNA structure. Furthermore, small angle X-ray scattering and circular dichroism studies indicate that the aggregate-enhancing N-terminal region of BR120-166 adopts an extended, non-globular structure. Altogether, our results suggest that PsrP adheres to extracellular DNA in the biofilm matrix and thus promotes pneumococcal biofilm formation.


Accidental coadministration of intravenous aminophylline and theophylline by mouth: a hazardous practice.
Eight patients who inadvertently received theophylline by mouth and aminophylline by the intravenous route together were reviewed by the Clinical Pharmacology Service of the Royal Adelaide Hospital because their serum theophylline levels were in excess of the stated therapeutic range. Six of these patients were severely compromised by their high theophylline level, and two of them, who had advanced obstructive lung disease and respiratory failure, died shortly after the drug levels were detected. Oversights of this nature can arise during the early stages of a patient's hospital admission and when medical staff changes occur. All patients who receive aminophylline by the intravenous route must be kept under close surveillance to ensure that theophylline formulations are not given by mouth concurrently.


Deaf Children's ASL Vocabulary and ASL Syntax Knowledge Supports English Knowledge.
The current study contributes empirical data to our understanding of how knowledge of American Sign Language (ASL) syntax aids reading print English for deaf children who are bilingual and bimodal in ASL and English print. The first analysis, a conceptual replication of Hoffmeister ( 2000), showed that performance on the American Sign Language Assessment Instrument correlated with the Sanford Achievement Test-Reading Comprehension (SAT-RC) and the Rhode Island Test of Language Structures (RITLS, Engen & Engen, 1983). The second analysis was a quantile regression using ASL assessments to predict English print abilities. Different ASL skills were important for English reading comprehension (SAT-RC) versus understanding English syntax (RITLS); the relationship between ASL skills and English print performance also varied for students at different English print ability levels. Strikingly, knowledge of ASL syntax was robustly correlated with knowledge of English syntax at all ability levels. Our findings provide novel and strong evidence for the impact of ASL on the development of English literacy.


Liver disease in pregnancy.
Acute viral hepatitis is the most common cause of jaundice in pregnancy. The course of acute hepatitis is unaffected by pregnancy, except in patients with hepatitis E and disseminated herpes simplex infections, in which maternal and fetal mortality rates are significantly increased. Chronic hepatitis B or C infections may be transmitted to neonates; however, hepatitis B virus transmission is effectively prevented with perinatal hepatitis B vaccination and prophylaxis with hepatitis B immune globulin. Cholelithiasis occurs in 6 percent of pregnancies; complications can safely be treated with surgery. Women with chronic liver disease or cirrhosis exhibit a higher risk of fetal loss during pregnancy. Preeclampsia is associated with HELLP (hemolysis, elevated liver enzymes and low platelet count) syndrome, acute fatty liver of pregnancy, and hepatic infarction and rupture. These rare diseases result in increased maternal and fetal mortality. Treatment involves prompt delivery, whereupon the liver disease quickly reverses. Therapy with penicillamine, trientine, prednisone or azathioprine can be safely continued during pregnancy.


Norovirus foodborne outbreaks associated with the consumption of oysters from the Etang de Thau, France, December 2002.
In January 2003, the Institut de Veille Sanitaire received notification of clusters of gastroenteritis (GE) thought to be associated with consumption of oysters harvested from Etang de Thau in the south of France. At the same time Italy reported an outbreak (200+ cases) associated with oysters from the Etang de Thau. An investigation was carried out to determine the source and vehicle of the outbreaks. Descriptive analysis of reported clusters in France, microbiological analysis of stool and oyster samples, genotyping of noroviruses and an environmental investigation of the Etang de Thau were carried out. A retrospective cohort study was also undertaken among those attending a number of family meals in Paris. Thirteen family clusters in four districts of France (69 cases) could be attributed to the consumption of Thau oysters based on descriptive evidence. Oysters distributed at an office in Paris and consumed at fourteen family meals between 19 and 24 December led to a further outbreak. In this outbreak the attack rate was 21/36 (58%) for Thau oyster consumers and 0/22 for non-consumers (p=0.00002). Noroviruses (genogroups I and II) were found in stool samples from four clusters and oysters from three clusters (including Paris). Environmental investigations revealed heavy rainfall, an overflow of a water purification station and faecal contamination of the Etang de Thau in December. Oysters from the Etang de Thau were responsible for a number of clusters of norovirus GE in winter 2002 in France and also in Italy. High Escherichia Coli levels in Thau water and shellfish led to an official request, mid-December, for oyster purification before distribution. This was not possible, due to lack of purification facilities. This investigation has contributed to a change in the way that shellfish harvesting areas are classified in France.


Confounding effect of EEG implantation surgery: Inadequacy of surgical control in a two hit model of temporal lobe epilepsy.
In rodent models of epilepsy, EEG implantation surgery is an essential modality to evaluate electrographic seizures. The inflammatory consequences of EEG electrode-implantation and their resultant effects on seizure susceptibility are unclear. We evaluated electrode-implantation in a two-hit model of epileptogenesis in C57BL/6 mice that included brief, recurrent febrile seizures (FS) at P14 and kainic acid induced seizures (KA-SZ) at P28. During KA-SZ, latencies to first electrographic and behavioral seizures, seizure severity, and KA dose sensitivity were measured. Mice that received subdural screw electrode implants at P25 for EEG monitoring at P28 had significantly shorter latencies to seizures than sham mice, regardless of early life seizure experience. Electrode-implanted mice were sensitive to low dose KA as shown by high mortality rate at KA doses above 10mg/kg. We then directly compared electrode-implantation and KA-SZ in seizure naive CX3CR1(GFP/+) transgenic C57BL/6 mice, wherein microglia express green fluorescent protein (GFP), to determine if microglia activation related to surgery was associated with the increased seizure susceptibility in electrode-implanted mice from the two-hit model. Hippocampal microglia activation, as demonstrated by percent area GFP signal and GFP positive cell counts, prior to seizures was indistinguishable between electrode-implanted mice and controls, but was significantly greater in electrode-implanted mice following seizures. Electrode-implantation had a confounding priming effect on the inflammatory response to subsequent seizures.


Proteoglycans and calcification of cartilage in the femoral head epiphysis of the immature rat.
The femoral heads of young rats have been used to monitor changes in proteoglycan structure during growth and endochondral ossification. Proteoglycans were extracted in good yield. The tissue content of proteoglycans increased until the time of calcification and then decreased. In contrast, the collagen content increased over the period studied. On Day 20, just preceding the onset of calcification, the proteoglycans had a lower glycosaminoglycan content, were somewhat smaller, and contained a larger proportion of molecules that were not capable of interacting with hyaluronic acid. On Day 25, during ongoing calcification, the proportion of proteoglycans that were not capable of interacting with hyaluronic acid was low, while it again was high on Day 40, just preceding ossification. The relative glycosaminoglycan content of the proteoglycans was somewhat lower on Day 20 and Day 40. The results indicate that both at the time of calcification and at the time of ossification the proteoglycan structure changes, perhaps indicating a functional role for the proteoglycans in the calcification process.


Practical Guidance for Involving Stakeholders in Health Research.
Stakeholder engagement is increasingly common in health research, with protocols for engaging multiple stakeholder groups becoming normative in patient-centered outcomes research. Previous work has focused on identifying relevant stakeholder groups with whom to work and on working with stakeholders in evidence implementation. This paper draws on the expertise of a team from four countries-Canada, Australia, the UK, and the USA-to provide researchers with practical guidance for carrying out multi-stakeholder-engaged projects: we present a list of questions to assist in selecting appropriate roles and modes of engagement; we introduce a matrix to help summarize engagement activities; and we provide a list of online resources. This guidance, matrix, and list of resources can assist researchers to consider more systematically which stakeholder groups to involve, in what study roles, and by what modes of engagement. By documenting how stakeholders are paired up with specific roles, the matrix also provides a potential structure for evaluating the impact of stakeholder engagement.


MiR-184-5p represses neuropathic pain by regulating CCL1/CCR8 signaling interplay in the spinal cord in diabetic mice.
Diabetic neuropathic pain (DNP) is a serious complication for diabetic patients involving nervous system. MicroRNAs (miRNAs) are small-noncoding RNAs which are dysregulated in neuropathic pain, and might be critical molecules for pain treatment. Our previous study has shown miR-184-5p was significantly downregulated in DNP. Therefore, the mechanism of miR-184-5p in DNP was investigated in this study. A DNP model was established through streptozotocin (STZ). The pharmacological tools were injected intrathecally, and pain behavior was evaluated by paw withdrawal mechanical thresholds (PWMTs). Bioinformatics analysis, Dual-luciferase reporter assay and fluorescence-in-situ-hybridization (FISH) were used to seek and confirm the potential target genes of miR-184-5p. The expression of relative genes and proteins was analyzed by quantitative reverse transcriptase real-time PCR (qPCR) and western blotting. MiR-184-5p expression was down-regulated in spinal dorsal on days 7 and 14 after STZ, while intrathecal administration of miR-184-5p agomir attenuates neuropathic pain induced by DNP and intrathecal miR-184-5p antagomir induces pain behaviors in naïve mice. Chemokine CC motif ligand 1 (CCL1) was found to be a potential target of miR-184-5p and the protein expression of CCL1 and the mRNA expression of CCR8 were up-regulated in spinal dorsal on days 7 and 14 after STZ. The luciferase reporter assay and FISH demonstrated that CCL1 is a direct target of miR-184-5p. MiR-184-5p overexpression attenuated the expression of CCL1/CCR8 in DNP; intrathecal miR-184-5p antagomir increased the expression of CCL1/CCR8 in spinal dorsal of naïve mice. This research illustrates that miR-184-5p alleviates DNP through the inhibition of CCL1/CCR8 signaling expression.


The influence of an overactive bladder on falling: a study of females aged 40 and older in the community.
An overactive bladder (OAB) affects a person's quality of life. Patients who suffer from OAB run to the toilet frequently to prevent incontinence, and this behavior increases their risk of falling and fear of falling. This study evaluated the influence of OAB on falls and concern about falling in females aged 40 and over living in urban and rural communities. We conducted a population-based cohort study using King's Health Questionnaire (KHQ), the Korean version of Falls Efficacy Scale-International (KFES-I) and a questionnaire regarding falls, in females aged 40 and over in Guri city and Yangpyeong county. The data from 514 responders were analyzed. The definition of OAB was 'moderately' or 'a lot' of urgency, or urge incontinence in KHQ. Falls was defined as experience of falls in the last year. High fear of falling was defined as a score of 24 or over in KFES-I. The factors were analyzed by the exact chi-square test and Student's t-test. The multivariate logistic regression model was adopted in order to examine the effects of OAB on falls and concern about falling. Of the 514 responders, 98 fitted the criterion of OAB. Eighty-nine (17.3%) of the responders had experienced falls in the last year: twenty-seven (27.5%) in the group with OAB and 62 (14.9%) in the group without OAB. There was a significant association between falls and OAB (odds ratio [OR], 1.76; 95% confidence interval [CI], 1.00 to 3.08; P=0.0485), and between high fear of falling and OAB (OR, 2.72; 95% CI, 1.42 to 5.20; P=0.0024). Urgency and symptoms of urge incontinence increase the risk of falls in women aged 40 or older in the community. Early diagnosis and proper treatment may prevent falls and improve quality of life in OAB patients.


Effect of pH Variations on the Yield Stress of Calcium Bentonite Slurry Treated with pH-Responsive Polymer.
The pH-responsive polymers, such as polyacrylamide (PAM), show distinct conformational states according to the pH of their environmental groundwater. Therefore, the interactions between clay-polymer and polymer-water molecules, which determine the yield stress of bentonite-polymer composites, can be affected by the pH of groundwater. This study aims to evaluate the effect of pH variation on the yield stress of calcium bentonite treated with PAM. The yield stresses (τy) of untreated and PAM treated clays were measured with varying volume fractions of solid (VF = 10-23%) and under varying pH conditions (pH = 7.6-9.6). In addition, the zeta potential was measured for both untreated and treated clays to figure out the change in the surface charge of the mineral surface due to PAM treatment. The results of this study demonstrate that τy for treated clay is higher than that for untreated clay at a given VF, because van der Walls attraction dominates electrostatic repulsion in the case of treated clay. Due to the change in conformational states of PAM and the consequent change in surface charge that comes with varying pH, the pH-dependent change in τy of treated clay is significantly different from that of untreated clay.


Platinoid effects on human plasmatic coagulation kinetics: a viscoelastic analysis.
In recent years a variety of metals (cadmium, chromium, copper, iron) have been demonstrated to modulate coagulation in vitro and in vivo. One group of metals, the platinoids, have not been assessed, and such investigation is justified given the thromboembolic phenomena associated with platinum-based chemotherapy. Thus, the goal of the present investigation was to assess the effects of carboplatin, cisplatin (platinum compounds), NAMI-A, and ruthenium chloride (ruthenium compounds) on human plasmatic coagulation. Human plasma was exposed to clinically relevant, equimolar concentrations of the aforementioned platinum and ruthenium compounds, with changes in plasmatic coagulation assessed via thrombelastography. The first series of experiments demonstrated no significant modulation of coagulation by the platinum compounds, while NAMI-A demonstrated mild hypercoagulability and ruthenium chloride exerted marked hypercoagulability. A second series of experiments utilizing a variety of specialized modifications of thrombelastography focused on ruthenium chloride revealed that this compound enhances prothrombin activation. While the hypercoagulability associated with platinum compounds in vivo do not appear to have a basis in plasmatic biochemistry, it appears that ruthenium compounds can exert procoagulant properties by enhancing the common pathway of human plasmatic coagulation. Future investigation of Ru based chemotherapeutic agents in development to assess procoagulant activity as part of evaluating their potential clinical safety is warranted.


Safety of right heart catheterization for pulmonary hypertension in very elderly patients.
Right heart catheterization (RHC) is the reference test in diagnosing pulmonary hypertension (PH). The increasing age of patients at the time of diagnosis raises the issue of the morbidity of this invasive test in elderly individuals. We hypothesized that the morbidity associated with RHC would be increased in elderly patients and highlight differences in hemodynamic characteristics compared to younger patients. A retrospective study was conducted in a regional referral center for PH. Data for all consecutive RHCs performed during the study period were analyzed. Over a five-year period, 1060 RHCs were performed. Of the patients, 228 (21.5%) were aged ≥75 years and 832 (78.5%) were aged <75 years. Duration of the procedure and site of puncture did not differ according to age group (all P > 0.05). Nine procedures (0.9%) led to complications: three (1.3%) in patients aged >75 years and six (0.7%) in younger patients aged ( P = 0.5). Eight were local vascular injuries, directly related to a femoral vein puncture ( P < 0.001). Pulmonary arterial pressure and cardiac output were lower in patients aged >75 years than in younger patients ( P = 0.001). RHC may be performed regardless of patient age. The rate of RHC complications is not increased in individuals aged >75 years. As most complications were related to femoral vein puncture, this route should be avoided whenever possible.


Imipramine is effective in preventing relapse in electroconvulsive therapy-responsive depressed inpatients with prior pharmacotherapy treatment failure: a randomized, placebo-controlled trial.
To compare the efficacy of imipramine versus placebo in preventing relapse after successful electroconvulsive therapy (ECT) in depressive inpatients with pharmacotherapy treatment failure prior to ECT. During a 6-month period, the incidence of relapse was assessed. Two centers, both inpatient units for treatment of depressed patients, participated in this trial. Patients with DSM-IV-diagnosed major depressive disorder resistant to an anti-depressant and subsequent lithium addition and/or a monoamine oxidase inhibitor were included. Patients were randomly assigned to double-blind treatment with imipramine with adequate plasma levels (N=12) or placebo (N=15) after successful ECT. The mean imipramine dosage was 209 mg/day (standard deviation: 91.7, range: 75-325 mg/day). The main outcome measure was relapse defined as at least "moderately worse" compared with baseline score on the Clinical Global Impressions-Improvement scale. Treatments were compared with survival analysis using the Cox proportional hazards model, including psychotic features and the score on the Hamilton Rating Scale for Depression (HAM-D) at baseline as prespecified covariables. Patients were enrolled in the study from April 1997 to July 2001. In the placebo group, 80% (12/15) of the patients relapsed compared with 18% (2/11) in the imipramine group. The Cox regression analysis showed a significant reduction in the risk of relapse of 85.6% with imipramine compared to placebo (p=.007; 95% confidence interval [CI]=24.6% to 97.2%) adjusted for the covariables. There was an 18% increase in the relapse rate (p=.032; 95% CI=2% to 36%) per unit increase in HAM-D score before the start of the trial; psychotic features had no significant effect (p=.794). Depressed patients with pharmacotherapy treatment failure may benefit from the prophylactic effect of the same class of drug during maintenance therapy after response to ECT.


Mother's occupation and sex ratio at birth.
Many women are working outside of the home, occupying a multitude of jobs with varying degrees of responsibilities and levels of psychological stress. We investigated whether different job types in women are associated with child sex at birth, with the hypothesis that women in job types, which are categorized as "high psychological stress" jobs, would be more likely to give birth to a daughter than a son, as females are less vulnerable to unfavourable conditions during conception, pregnancy and after parturition, and are less costly to carry to term. We investigated the effects of mother's age, maternal and paternal job type (and associated psychological stress levels) and paternal income on sex ratio at birth. Our analyses were based on 16,384 incidences of birth from a six-year (2000 to 2005 inclusive) childbirth dataset from Addenbrooke's Hospital in Cambridge, UK. We obtained a restricted data set from Addenbrooke's hospital with: maternal age, maternal and paternal occupations, and whether or not the child was first-born. Women in job types that were categorized as "high stress" were more likely to give birth to daughters, whereas women in job types that were categorized as "low stress" had equal sex ratios or a slight male bias in offspring. We also investigated whether maternal age, and her partner's income could be associated with reversed offspring sex ratio. We found no association between mother's age, her partner's job stress category or partner income on child sex. However, there was an important interaction between job stress category and partner income in some of the analyses. Partner income appears to attenuate the association between maternal job stress and sex ratios at moderate-income levels, and reverse it at high-income levels. To our knowledge this is the first report on the association between women's job type stress categories and offspring sex ratio in humans, and the potential mitigating effect of their partners' income.


[Phenolic extraction of nucleic acids from the spermatozoa of the roach and the bream].
The results are presented of nucleic acids isolation from spermatozoids of roach and bream by the method of nucleproteid phenol deproteinization. The quantitative yield is given for DNA and RNA preparations from the mentioned objects, the molecular weight of roach and bream DNA, nucleotide composition of roach DNA as well as fractional composition of bream spermatozoids RNA are determined.


Reformation of La0.7Sr0.3MnO3 properties by using ZnO in La0.7Sr0.3MnO3-ZnO heterostructures grown on (001) oriented Si.
Study of the available density of states (DOS) close-to-zero bias for conduction in strongly correlated electron systems, such as half-metallic La0.7Sr0.3MnO3 (LSMO) and its heterostructures, is important for fundamental and application reasons. As the DOS is proportional to the differential conductance (dI/dV), the dI/dV of a 120 Å LSMO film and its reformation in LSMO/ZnO heterostructures was investigated for different ZnO thicknesses. Unlike in conventional metals, the dI/dV of LSMO exhibits a power-law dependent zero-bias anomaly, i.e., dI/dV ∝ Vm (m ∼ 1) near zero bias in the ferromagnetic metallic state at 10 K. The growth of ZnO on LSMO reforms the linear dI/dVvs. V of LSMO near zero bias to non-linear. The exponent 'm' becomes ∼0.5 for a higher ZnO thickness, revealing increased electron-electron interactions and suppression of Kondo-like, double and superexchange interactions, which are responsible for the depression of the DOS of LSMO near zero bias. In a magnetically disordered state, i.e., around the Curie temperature, ZnO reforms the linear V-shaped dI/dV vs. V of LSMO to parabolic U-shaped dI/dVvs.V and controls the electron concentrations in the t2g-orbitals of Mn realized from the DOS simulations. Additionally, ZnO introduces a peak in the dI/dV vs. V due to Fowler-Nordheim tunnelling, and the peak voltage can be tuned by varying the ZnO thickness or temperature from 300 K to 360 K. Such functions of ZnO yield major perspectives for novel applications in thin-film-based devices.


Emergence of non-albicans Candida species in neonatal candidemia.
Candida species are one of the most common causes of blood stream infections among neonates and account for 9-13% of such infections. Although Candida albicans remains the most common fungal isolate from neonatal candidemia, longitudinal studies have detected a shift towards non-albicans Candida (NAC) species. To examine the prevalence and epidemiology of candidemia among infants admitted to our hospital. Blood samples were collected from 548 neonates and only those which yielded pure growth of Candida spp. were included in the study. The isolates were identified as per standard mycological techniques and antifungal susceptibility (AFS) was done by disc diffusion method. Of the total 132 neonates included in the study, NAC species were responsible for 80.30% cases with C. parapsilosis (25.0%) and C. tropicalis (21.97%) as the most predominant species; whereas 19.70% of cases were caused by C. albicans. AFS results revealed that 65.91, 73.49, and 96.21% isolates were sensitive to fluconazole (FLK), itraconazole (ITR), and amphotericin B (AMB), respectively. Candidemia in neonates is an ominous prognostic sign and is an important entity in our hospital. Strict infection control strategies, appropriate preventive and therapeutic measures such as prophylactic antifungal use and a restrictive policy of antibiotic use should be implemented.


Reduction of stutter ratios in short tandem repeat loci typing of low copy number DNA samples.
Increased height of stutter peaks is a phenomenon with low copy number (LCN) short tandem repeat (STR) typing that can impact interpretation. An alternative strategy of lowering the annealing/extension temperature (LT) at 56 °C was designed to attempt to decrease the heights of stutter peaks. STR typing results were generated in terms of stutter ratios using LT-PCR conditions and compared with data obtained using standard (STD) PCR conditions. DNA samples ranging from 100 to 25 pg were amplified using reagents contained in the AmpFℓSTR Identifiler PCR Amplification or AmpFℓSTR Identifiler Plus PCR Amplification kits with 32 or 34 PCR cycles. Stutter ratios decreased by an average of 14.7%, 14.9% and 18.1% at 100, 50 and 25 pg of template DNA under LT conditions compared with those of STD conditions in the Identifiler Kit amplified samples. The LT conditions also decreased average stutter ratios by 13.3% compared with those of STD conditions in the Identifiler Plus Kit amplified samples. The overall PCR efficiency obtained with STD and LT conditions with the two STR kits was comparable in terms of the number of detected alleles, peak heights and peak height ratios. These results support the hypothesis that a lower temperature annealing/extension step reduces the likelihood of slippage during PCR by enhancing the stability of the DNA polymerase/template DNA complex or the stability of the generated duplex than the conditions of the standard extension step. This stability in turn would result in lower stutter ratios.


[Signal processing in ultrasound imaging, Doppler and Doppler imaging].
The main purpose of signal processing in ultrasound imaging is the improvement of the image quality. The basics of three-dimensional, nonlinear image filtering are described in a simple way. For Doppler several million operations are computed per second to obtain the velocity distribution as a function of time. Various methods are described for overcoming the alias problem. For Doppler imaging the necessary computing power increases to several hundred million operations per second. Like other medical imaging methods, diagnostic ultrasound depends on powerful electronic computers.


Diastereoselective Johnson-Corey-Chaykovsky trifluoroethylidenation.
(2,2,2-Trifluoroethyl)diphenylsulfonium triflate was found to be an efficient ylide reagent for the Johnson-Corey-Chaykovsky reaction to afford trifluoromethyl epoxides, cyclopropanes and aziridines. Interestingly, excellent but different diastereoselectivity was observed for these transformations. Both trifluoromethyl epoxides and cyclopropanes were obtained with trans-selectivity, whereas aziridines were obtained with cis-selectivity.


Detection of anti-HAV antibody with dot immunogold filtration assay.
To establish a rapid, sensitive and specific immunogold assay for detection of hepatitis A virus infection. Rabbit monoclonal antibodies to anti-human IgM and IgG (Dako) were dotted on a nitrocellulose membrane (NCM) respectively to capture the human sera IgM and IgG. Then the captured antibodies would conjugate to HAV antigen, which was revealed by mouse anti-HAV IgG conjugated to gold particles. Final results were assessed by blind method. Sera from 96 patients with acute hepatitis were used for our study. Compared with well-recognized standard (Abbott Laboratory, USA), the sensitivity and specificity of IgM-DIGFA (self-made) were 91.3 % (42/46) and 96.0 % (48/50), and those of IgM-ELISA (Kehua, Shanghai) were 97.8 % (45/46) and 100.0 % (50/50). The identical results were produced from the study with reagents at different conditions, and the study was repeated in 15 negative sera and 10 positive sera. The serum anti-HAV IgG was tested with DIGFA at the same time. In comparison with ELISA, the sensitivity and specificity of DIGFA for IgG anti-HAV were 87.2 % (41/47) and 91.8 % (45/49), respectively. This assay can detect anti-HAV IgM and IgG simultaneously, and be done within 3 minutes. The simplicity, rapidity and specificity of the assay were useful for screening and epidemiological study.


PTGS2 (COX-2) -765G > C promoter variant reduces risk of colorectal adenoma among nonusers of nonsteroidal anti-inflammatory drugs.
Prostaglandin H synthase 2 (PTGS2) or cyclooxygenase-2 (COX-2) has been shown to play a key role in the regulation of inflammation, and its inhibition is associated with a reduced risk of colon cancer. The PTGS2 (COX-2) -765G > C promoter variant is located in a putative SP1 binding site and reduces PTGS2 expression. In a Minnesota-based case-control study of cases with adenomatous (n = 494) or hyperplastic polyps (n = 186) versus polyp-free controls (n = 584), we investigated the role of the PTGS2 -765G > C promoter polymorphism. Multiple logistic regression analysis was used, adjusting for age, body mass index, caloric intake, alcohol, fiber, sex, hormone use, and smoking. For colorectal adenoma, odds ratios (OR) compared with PTGS2 -765GG as reference were GC 1.00 [95% confidence interval (95% CI), 0.74-1.35] and CC 0.53 (95% CI, 0.22-1.28). For hyperplastic polyps, the comparable adjusted odds ratios were GC 0.97 (95% CI, 0.65-1.46) and CC 0.24 (95% CI, 0.05-1.11). Risk associated with the -765G > C variant differed by aspirin or other nonsteroidal anti-inflammatory drug (NSAID) use. Among nonusers of aspirin or other NSAIDs, the CC genotype conferred a significant decrease in risk of adenoma (OR, 0.26; 95% CI, 0.07-0.89). Use of aspirin or other NSAIDs reduced risk of adenoma only among those with the -765GG (wild type) and possibly -765CG genotypes (OR, 0.66; 95% CI, 0.48-0.92 and OR, 0.64; 95% CI, 0.40-1.02, respectively). These data suggest that COX-2 expression or activity may be beneficially suppressed, and risk of colorectal polyps reduced, by aspirin or other NSAIDs in PTGS2 -765GG (wild type) individuals and by the -765 CC variant genotype in nonusers of NSAIDs.


Intravascular hemolysis, vascular endothelial cell activation and thrombophilia in splenectomized patients with hemoglobin E/β-thalassemia disease.
The relationship between asplenia and thrombophilia in β-thalassemia disease patients is not yet completely understood. One hundred and ten adult hemoglobin (Hb) E/β-thalassemia (E/β-Thal) disease outpatients, dichotomized according to the presence or absence of the spleen, were prospectively studied for evidence of intravascular hemolysis (IVH) and vascular endothelial cell (EC) activation. Biomarkers of IVH (serum cell-free Hb), EC [soluble E-selectin (sE-selectin) and soluble vascular cell adhesion molecule 1 (sVCAM-1)], platelet and EC [soluble P-selectin (sP-selectin)], inflammation [high-sensitivity C-reactive protein (hs-CRP)], and coagulation [thrombin-antithrombin complexes (TAT)] activation, as well as other selected blood tests were determined. The 61 splenectomized patients had a more severe hemolytic disease and higher levels of cell-free Hb and ferritin (p = 0.003), sE-selectin, sP-selectin, hs-CRP, and TAT (p < 0.05). However, serum levels of sVCAM-1 were not different between the two groups. The findings suggested IVH and EC activation. Together with chronic iron overload and chronic low-grade inflammation activation, the findings extend our understanding of the mechanism of thrombophilia in splenectomized E/β-Thal disease patients.


Abnormal cardiac adrenergic nerve function in patients with syndrome X detected by [123I]metaiodobenzylguanidine myocardial scintigraphy.
Previous studies have suggested that an abnormal cardiac adrenergic tone may have a pathophysiological role in syndrome X (effort angina, positive exercise testing, angiographically normal coronary arteries). To evaluate cardiac adrenergic nerve function, we performed [123I]metaiodobenzylguanidine (MIBG) myocardial scintigraphy in 12 patients with syndrome X and 10 control subjects. Cardiac MIBG uptake was assessed by the heart/mediastinum (H/M) ratio and by an MIBG uptake defect score (higher values=lower uptake). In syndrome X patients, we also correlated MIBG scintigraphic findings with stress myocardial perfusion as assessed by 201Tl scintigraphy. An inferior MIBG defect was observed in only 1 control subject, whereas 9 patients (P<.01) showed MIBG defects. The heart was totally or almost totally invisible on MIBG images in 5 patients, and predominantly regional defects were observed in 4. The H/M ratio was lower (1.70+/-0.6 versus 2.2+/-0.3, P=.03) and MIBG uptake defect score higher (35+/-31 versus 4+/-2, P=.003) in syndrome X patients. Reversible stress thallium perfusion defects were found in 62% of patients with MIBG defects but in no patient with normal MIBG uptake. MIBG defects persisted unchanged in 7 patients at a 5+/-3-month follow-up study. In this study, obvious defects in global and/or regional cardiac MIBG uptake, indicating an abnormal cardiac adrenergic nerve function, were detected in 75% of patients with syndrome X. These findings strongly support the cardiac origin of chest pain in syndrome X, although the mechanisms and the pathophysiological meaning of the abnormal cardiac MIBG uptake in these patients deserve further investigation.


Imaging Mass Spectrometry and Proteome Analysis of Marek's Disease Virus-Induced Tumors.
The highly oncogenic alphaherpesvirus Marek's disease virus (MDV) causes immense economic losses in the poultry industry. MDV induces a variety of symptoms in infected chickens, including neurological disorders and immunosuppression. Most notably, MDV induces transformation of lymphocytes, leading to T cell lymphomas in visceral organs with a mortality of up to 100%. While several factors involved in MDV tumorigenesis have been identified, the transformation process and tumor composition remain poorly understood. Here we developed an imaging mass spectrometry (IMS) approach that allows sensitive visualization of MDV-induced lymphoma with a specific mass profile and precise differentiation from the surrounding tissue. To identify potential tumor markers in tumors derived from a very virulent wild-type virus and a telomerase RNA-deficient mutant, we performed laser capture microdissection (LCM) and thereby obtained tumor samples with no or minimal contamination from surrounding nontumor tissue. The proteomes of the LCM samples were subsequently analyzed by quantitative mass spectrometry based on stable isotope labeling. Several proteins, like interferon gamma-inducible protein 30 and a 70-kDa heat shock protein, were identified that are differentially expressed in tumor tissue compared to surrounding tissue and naive T cells. Taken together, our results demonstrate for the first time that MDV-induced tumors can be visualized using IMS, and we identified potential MDV tumor markers by analyzing the proteomes of virus-induced tumors.IMPORTANCE Marek's disease virus (MDV) is an oncogenic alphaherpesvirus that infects chickens and causes the most frequent clinically diagnosed cancer in the animal kingdom. Not only is MDV an important pathogen that threatens the poultry industry but it is also used as a natural virus-host model for herpesvirus-induced tumor formation. In order to visualize MDV-induced lymphoma and to identify potential biomarkers in an unbiased approach, we performed imaging mass spectrometry (IMS) and noncontact laser capture microdissection. This study provides a first description of the visualization of MDV-induced tumors by IMS that could be applied also for diagnostic purposes. In addition, we identified and validated potential biomarkers for MDV-induced tumors that could provide the basis for future research on pathogenesis and tumorigenesis of this malignancy.


Endotoxin-induced endothelial fibrosis is dependent on expression of transforming growth factors β1 and β2.
During endotoxemia-induced inflammatory disease, bacterial endotoxins circulate in the bloodstream and interact with endothelial cells (ECs), inducing dysfunction of the ECs. We previously reported that endotoxins induce the conversion of ECs into activated fibroblasts. Through endotoxin-induced endothelial fibrosis, ECs change their morphology and their protein expression pattern, thereby suppressing endothelial markers and upregulating fibrotic proteins. The most commonly used fibrotic inducers are transforming growth factor β1 (TGF-β1) and TGF-β2. However, whether TGF-β1 and TGF-β2 participate in endotoxin-induced endothelial fibrosis remains unknown. We have shown that the endotoxin-induced endothelial fibrosis process is dependent on the TGF-β receptor, ALK5, and the activation of Smad3, a protein that is activated by ALK5 activation, thus suggesting that endotoxin elicits TGF-β production to mediate endotoxin-induced endothelial fibrosis. Therefore, we investigated the dependence of endotoxin-induced endothelial fibrosis on the expression of TGF-β1 and TGF-β2. Endotoxin-treated ECs induced the expression and secretion of TGF-β1 and TGF-β2. TGF-β1 and TGF-β2 downregulation inhibited the endotoxin-induced changes in the endothelial marker VE-cadherin and in the fibrotic proteins α-SMA and fibronectin. Thus, endotoxin induces the production of TGF-β1 and TGF-β2 as a mechanism to promote endotoxin-induced endothelial fibrosis. To the best of our knowledge, this is the first report showing that endotoxin induces endothelial fibrosis via TGF-β secretion, which represents an emerging source of vascular dysfunction. These findings contribute to understanding the molecular mechanism of endotoxin-induced endothelial fibrosis, which could be useful in the treatment of inflammatory diseases.


Considerable pancreatic tumor motion during breath-holding.
Breath-holding (BH) is often used to reduce abdominal organ motion during radiotherapy. However, for inhale BH, abdominal tumor motion during BH has not yet been investigated. The aim of this study was to quantify tumor motion during inhale BH and tumor position variations between consecutive inhale BHs in pancreatic cancer patients. Twelve patients with intratumoral fiducials were included and asked to perform three consecutive 30-second inhale BHs on each of three measurement days. During BH, lateral fluoroscopic movies were obtained and a two-dimensional (2D) image correlation algorithm was used to track the fiducials and the diaphragm, yielding the tumor and diaphragm motion during each BH. The tumor position variation between consecutive BHs was obtained from the difference in initial tumor position between consecutive BHs on a single measurement day. We observed tumor motion during BH with a mean absolute maximum displacement over all BHs of 4.2 mm (range 1.0-11.0 mm) in inferior-superior (IS) direction and 2.7 mm (range 0.5-8.0 mm) in anterior-posterior (AP) direction. We found only a moderate correlation between tumor and diaphragm motion in the IS direction (Pearson's correlation coefficient |r|>0.6 in 45 of 76 BHs). The mean tumor position variation between consecutive BHs was 0.2 [standard deviation (SD) 1.7] mm in the inferior direction and 0.5 (SD 0.8) mm in the anterior direction. We observed substantial pancreatic tumor motion during BH as well as considerable position variation between consecutive BHs on a single day. We recommend further quantifying these uncertainties before introducing breath-hold during radiation treatment of pancreatic cancer patients. Also, the diaphragm cannot be used as a surrogate for pancreatic tumor motion.


Innate Immunity and Pathogenesis of Biliary Atresia.
Biliary atresia (BA) is a devastating fibro-inflammatory disease characterized by the obstruction of extrahepatic and intrahepatic bile ducts in infants that can have fatal consequences, when not treated in a timely manner. It is the most common indication of pediatric liver transplantation worldwide and the development of new therapies, to alleviate the need of surgical intervention, has been hindered due to its complexity and lack of understanding of the disease pathogenesis. For that reason, significant efforts have been made toward the development of experimental models and strategies to understand the etiology and disease mechanisms and to identify novel therapeutic targets. The only characterized model of BA, using a Rhesus Rotavirus Type A infection of newborn BALB/c mice, has enabled the identification of key cellular and molecular targets involved in epithelial injury and duct obstruction. However, the establishment of an unleashed chronic inflammation followed by a progressive pathological wound healing process remains poorly understood. Like T cells, macrophages can adopt different functional programs [pro-inflammatory (M1) and resolutive (M2) macrophages] and influence the surrounding cytokine environment and the cell response to injury. In this review, we provide an overview of the immunopathogenesis of BA, discuss the implication of innate immunity in the disease pathogenesis and highlight their suitability as therapeutic targets.


Autologous chondrocyte implantation using the original periosteum-cover technique versus matrix-associated autologous chondrocyte implantation: a randomized clinical trial.
Autologous chondrocyte implantation (ACI) is frequently used to treat symptomatic defects of the articular cartilage. To test whether matrix-associated autologous chondrocyte implantation or the original periosteal flap technique provides superior outcomes in terms of clinical efficacy and safety. Randomized controlled trial; Level of evidence, 2. Twenty-one patients (mean age, 29.3 +/- 9.1 years) with symptomatic isolated full-thickness cartilage defects (mean 4.1 +/- 09 cm2) at the femoral condyle were randomized to matrix-associated autologous chondrocyte implantation or the original periosteal flap technique. The primary outcome parameter was the postoperative change in knee function as assessed by the International Knee Documentation Committee (IKDC) score at 12 months after ACI. In addition, the IKDC score was assessed at 3, 6, 12, and 24 months after surgery. Secondary outcome parameters were postoperative changes in health related quality of life (Short Form-36 Health Survey), knee functionality (Lysholm and Gillquist score), and physical activity (Tegner Activity Score) at 3, 6, 12, and 24 months after ACI. Magnetic resonance imaging was performed to evaluate the cartilage 6, 12, and 24 months after ACI and rated using the Magnetic Resonance Observation of Cartilage Repair Tissue score. Adverse events were recorded to assess safety. The primary outcome parameter showed improvement of patients 1 year after autologous chondrocyte implantation, but there was no difference between the periosteal flap technique and matrix-associated ACI (P = .5573); 2 years after ACI, a similar result was found (P = .4994). The study groups did not show differences in the Short Form-36 categories and in knee functionality as assessed by Tegner Activity Score 12 months (P = .4063) and 24 months (P = .1043) after ACI. There was a significant difference in the Lysholm and Gillquist score at 12 months (P = .0449) and 24 months (P = .0487) favoring the periosteal flap technique group. At 6 months after surgery, a significantly lower Magnetic Resonance Observation of Cartilage Repair score was obtained in the matrix-associated ACI group (P = .0123), corresponding to more normal magnetic resonance imaging diagnostic findings. Twelve and 24 months after ACI, the differences between the 2 groups were not significant (12 months, P = .2065; 24 months, P = .6926). Adverse events were related to knee problems such as transplant delamination, development of an osseous spur, osteochondral dissection, and transplant hypertrophy. Systemic (allergic, toxic, or autoimmune) reactions did not occur. There was no difference in the efficacy between the original and the advanced ACI technique 12 and 24 months after surgery regarding International Knee Documentation Committee, Tegner Activity Score, and Short Form-36; however, with respect to the Lysholm and Gillquist score, better efficacy was observed in the periosteal flap technique group.


Diabetes and employment productivity: does diabetes management matter?
To determine whether labor market effects were the result of diabetes per se or rather depended on the degree to which diabetes was controlled through management of blood sugar levels. This study utilized data from a recently completed survey of households in Brownsville, Texas, a largely Mexican American community with a high prevalence of diabetes that is located on the Texas-Mexico border. Diabetes management, or control, was measured by blood sugar levels, glycosylated hemoglobin (A1C) levels, and interaction terms. Methods used were probit and Heckman regression. Management of diabetes did not appear to have a discernible impact on labor market outcomes in the short run. However, diabetes was negatively associated with male productivity, particularly in males' propensity to work. The new American Diabetes Association (ADA) definition of diabetes is based on having an A1C level of <6.5%. Using the new ADA definition, diabetes was negatively associated with female productivity. Female productivity was also negatively associated with the new ADA definition of prediabetes (A1C levels between 5.7% and 6.4%). However, very few people with diabetes actually controlled their blood sugar or A1C levels. These results imply that in order to avoid productivity losses associated with diabetes, more scarce prevention resources should be spent on prevention of the onset of diabetes than on the management of A1C for those already diagnosed with diabetes. For women, the prevention of prediabetes is key.


Corticosterone: a critical factor in an opioid form of stress-induced analgesia.
The finding that some opioid-mediated forms of stress-induced analgesia are antagonized by hypophysectomy and dexamethasone has led to the suggestion that beta-endorphin, released from the pituitary, may mediate these analgesic reactions. "Long-term analgesia" (an opioid-mediated form of stress-induced analgesia), which is blocked by dexamethasone and hypophysectomy, was also blocked by adrenalectomy and reinstated with corticosterone therapy. Corticosterone is proposed to play a permissive role in long-term analgesia and to be a critical hormone mediating this phenomenon.


In Vitro Storage of Functional Sperm at Room Temperature in Zebrafish and Medaka.
The longevity of sperm in teleost such as zebrafish and medaka is short when isolated even in saline-balanced solution at a physiological temperature. In contrast, some internal fertilizers exhibit the long-term storage of sperm, >10 months, in the female reproductive tract. This evidence implies that sperm in teleost possesses the ability to survive for a long time under suitable conditions; however, these conditions are not well understood. In this study, we show that the sperm of zebrafish can survive and maintain fertility in L-15-based storage medium supplemented with bovine serum albumin, fetal bovine serum, glucose, and lactic acid for 28 days at room temperature. The fertilized embryos developed to normal fertile adults. This storage medium was effective in medaka sperm stored for 7 days at room temperature. These results suggest that sperm from external fertilizer zebrafish and medaka has the ability to survive for at least 4 and 1 week, respectively, in the body fluid-like medium at a physiological temperature. This sperm storage method allows researchers to ship sperm by low-cost methods and to investigate key factors for motility and fertile ability in those sperm.


BR22, a 26 kDa thyroid transcription factor-1 associated protein (TAP26), is expressed in human lung cells.
The current authors have previously identified BR22, a thyroid transcription factor (TTF)-1 associated protein 26 (TAP26), which interacts with TTF-1 to enhance human surfactant protein (SP)-B promoter activity in transfected 293 cells. However, the expression of TAP26 in the lung cells and its biological relevance to the SP-B production under physiological conditions were not examined. In this study, endogenous co-immunoprecipitation and in situ immunohistochemical staining techniques were employed to explore the presence of TAP26 and TTF-1 complex in the lung epithelial cells. The correlation of TAP26, TTF-1 and SP-B expression was inspected in H441 cells in the presence of dexamethasone, a known positive effector of the SP-B promoter. Monoclonal antibody (mAb) against TAP26 can co-immunoprecipitate both TAP26 and TTF-1 from H441 cells. Using this antibody in in situ staining of human lung sections, the data show that TAP26 is present in the lung alveolar epithelial cells. Reverse transcriptase-polymerase chain reaction and Western blot analyses of type-II cells as well as dexamethasone-treated H441 cells suggest that TAP26 expression is modulated coordinately with SP-B and TTF-1 in these cells. In summary, the current study demonstrates that thyroid transcription factor-1 associated protein 26 is an associated protein of thyroid transcription factor-1 in the lung alveolar epithelial cells where surfactant protein gene expressions take place in vivo.


MK3 modulation affects BMI1-dependent and independent cell cycle check-points.
Although the MK3 gene was originally found deleted in some cancers, it is highly expressed in others. The relevance of MK3 for oncogenesis is currently not clear. We recently reported that MK3 controls ERK activity via a negative feedback mechanism. This prompted us to investigate a potential role for MK3 in cell proliferation. We here show that overexpression of MK3 induces a proliferative arrest in normal diploid human fibroblasts, characterized by enhanced expression of replication stress- and senescence-associated markers. Surprisingly, MK3 depletion evokes similar senescence characteristics in the fibroblast model. We previously identified MK3 as a binding partner of Polycomb Repressive Complex 1 (PRC1) proteins. In the current study we show that MK3 overexpression results in reduced cellular EZH2 levels and concomitant loss of epigenetic H3K27me3-marking and PRC1/chromatin-occupation at the CDKN2A/INK4A locus. In agreement with this, the PRC1 oncoprotein BMI1, but not the PCR2 protein EZH2, bypasses MK3-induced senescence in fibroblasts and suppresses P16INK4A expression. In contrast, BMI1 does not rescue the MK3 loss-of-function phenotype, suggesting the involvement of multiple different checkpoints in gain and loss of MK3 function. Notably, MK3 ablation enhances proliferation in two different cancer cells. Finally, the fibroblast model was used to evaluate the effect of potential tumorigenic MK3 driver-mutations on cell proliferation and M/SAPK signaling imbalance. Taken together, our findings support a role for MK3 in control of proliferation and replicative life-span, in part through concerted action with BMI1, and suggest that the effect of MK3 modulation or mutation on M/SAPK signaling and, ultimately, proliferation, is cell context-dependent.


Total and respirable dust in swine confinement buildings: the benefit of respiratory protective masks and effect of recirculated air.
Caretakers and pigs in dusty environments inhale particles and toxic gases which can cause subclinical illness. We determined the reduction in sampled dust elicited by respiratory masks mounted on glass funnels. Open-faced filters or British cyclones were sampled to measure quantities of dust which the masks had trapped. Respiratory masks reduced the sampled total suspended particulates (open-faced filters) by > or = 75% with NIOSH/MSHA certified protectant (2-tie) masks and > or = 50% with NIOSH/MSHA non-certified comfort (1-tie) masks. Respirable particulates (British cyclones) were reduced by > or = 45% with NIOSH/MSHA certified protectant (2-tie) masks. These data suggest properly worn respiratory protective masks afford significant protection against both total suspended and respirable particulates in swine confinement facilities. Penetration of 3-25% of total aerosol mass through masks allowing only 1% penetration of a silica aerosol with an aerodynamic diameter of 0.6-1.0 microns suggested that measurable portions of the aerosol mass in these confinement houses behaved as if they were less than 1 micron in diameter. Because of the small size of the aerosol, NIOSH/MSHA certified respiratory protective masks should be worn when working in those facilities. We also studied concentrations of ammonia, endotoxin and total and respirable dust particles to determine effects of a recirculation fan which increased the nominal air flow capacity of the building by 10%. Recirculated air had minimal effects on ammonia, total airborne endotoxin or total particulate mass.(ABSTRACT TRUNCATED AT 250 WORDS)


Viewpoint: Central adjudication of myocardial infarction in outcome-driven clinical trials--common patterns in TRITON, RECORD, and PLATO?
Central adjudication in randomised controlled outcome-driven trials represents a traditional approach to maintain data integrity by applying uniformed rules for assessment of clinical events. It was the purpose of this investigation to determine the patterns of myocardial infarction (MI) adjudication in the TRITON, RECORD, and PLATO trials. We were matching centrally-adjudicated MI's (CAMI's) from the official trial publication with the site-reported MI (SRMI's) count from the Food and Drug Administration's secondary analyses for the investigational compounds prasugrel (TRITON), rosiglitazone (RECORD), and ticagrelor (PLATO). CAMI numbers showed a remarkable discrepancy to SRMI's by more than a doubling of the difference: from 72 to 145 events in TRITON favoring prasugrel (from a hazard ratio [HR]=0.76, p=0.08; to a HR=0.76, p<0.001), and from 44 to 89 events in favour of ticagrelor in PLATO (from a HR=0.94, p=0.095; to a HR=0.84, p<0.001). In contrast, in the RECORD trial, the CAMI count was less than the SRMI count (from 24 to 8 events, from a HR=1.42, p=0.93; to a HR=1.14, p=0.96), in this case diminishing cardiovascular hazards in favour of rosiglitazone. In conclusion, central adjudication in the TRITON, the RECORD, and the PLATO trial turned out to have a critical impact on study outcomes. Trial publications should in the future include site-reported major efficacy and safety endpoints to preserve data integrity. The regulatory authorities should consider independent audits when there is a major disagreement between centrally adjudicated and site reported events influencing the results of a major clinical trial.


Compact and efficient 3D shape description through radial function approximation.
A fast and simple method for three-dimensional shape description is described. The method views a 3D object as a radial distance function on the unit sphere, and thus reduces the dimensionality of the description problem by one. The radial distance function is approximated by Fourier methods in the basis of the spherical harmonic polynomials. The necessary integration is carried out on the object boundary, rather than on the unit sphere. Consequently, there is no need of a parameterisation of the object surface. The description makes it possible to compare shapes in a computationally very simple way. Solutions on how to cope with translated and rotated objects are discussed. The method is developed for star-shaped objects, but is stable even if the input image is non-star-shaped. The method is tested in a data set from magnetic resonance imaging (MRI) of the brain. Potential medical applications are discussed.


Outcome of HIV-1-associated cryptococcal meningitis, Denmark 1988-2008.
The risk of HIV-1-associated Cryptococcus neoformans meningitis (CM) has decreased and the outcome has improved with the use of combination antiretroviral therapy (cART). Outcome has not been reported in Denmark in the cART era. A review of all cases of HIV-1-associated CM treated at 2 hospitals in Denmark was carried out. Survival was compared by time-updated Cox proportional hazards analysis. A total of 45 cases were evaluated. Six individuals (13.3%) died within 30 days of being diagnosed with CM. cART was initiated a median of 15 days (range 3-53) after a diagnosis of CM for 12 individuals and did not affect 30-day outcome. Older age, however, was associated with an increased risk of death at 30 days (mortality rate ratio (MMR) 1.16 (95% confidence interval (95% CI) 1.05-1.30) per y increment). Twenty-four (55.8%) of 43 individuals (2 had emigrated) died within the 1(st) y. Initiation of cART significantly improved 1-y outcome (MMR 0.22, 95% CI 0.06-0.77). Mental status, CD4 T cell count, and antifungal did not affect short- or long-term outcome. We found that long-term survival after HIV-1-associated CM has improved significantly with the use of cART. Short-term mortality was not affected by initiation of cART and remained high.


Topical minoxidil in the treatment of alopecia areata.
A modified double blind crossover study was performed to assess the effect of 1% topical minoxidil as compared with placebo in 30 patients with alopecia areata and alopecia totalis. The active preparation produced a highly significant incidence of hair regrowth. A cosmetically acceptable response was noted in 16 patients. No side effects were seen. The study confirmed that topical minoxidil will induce new hair growth in alopecia areata but that it is less likely to do so in more severe and extensive disease. Furthermore, patients with alopecia universalis and totalis may not respond at all. Nevertheless, as compared with other drugs minoxidil applied topically is relatively non-toxic, is easy to use, and has no systemic or local side effects.


Formation of one or more intrachain disulphide bonds is required for the intracellular processing and transport of CD36.
In monocytes/macrophages, CD36 is thought to have a role as a scavenger receptor, mediating the phagocytosis of apoptotic cells and the endocytic uptake of oxidized low-density lipoproteins and fatty acids. The proposed topology of CD36 predicts that, of ten cysteine residues, six lie in the extracellular domain, whereas four are equally distributed in the two short terminal tails flanking the N-terminal and C-terminal hydrophobic stretches. Here we investigate the formation of intrachain disulphide bonds, on the basis of the assumption that the cysteine residues present in the luminal domains are generally oxidized, whereas those in the cytosol are reduced. As revealed by gel mobility-shift assays, disulphide bonds are present in the extracellular domain of the CD36 molecule. The formation of these bonds is required for the transport of CD36 from endoplasmic reticulum to Golgi. Furthermore reactive thiol groups are present in the CD36 sequence, which upon lysis form an intrachain extra loop as an artifact. This disulphide bond is not formed in either (1) truncated CD36 lacking the two C-terminal cysteine residues or (2) Triton X-100-insoluble wild-type CD36 molecules, suggesting that, in this fraction, the C-terminal thiol groups are modified.