Hormonal and developmental regulation of DAX-1 expression in Sertoli cells.
Mutations in the human DAX-1 gene lead to X-linked adrenal hypoplasia congenita and hypogonadotropic hypogonadism. DAX-1 has been proposed to play a role in steroidogenesis because it is highly expressed in adrenocortical and testicular Leydig cells and because loss-of-function mutations lead to low serum levels of steroid hormones. Recent reports of DAX-1 expression in hypothalamus and pituitary, however, suggest additional functions for this protein. Here we demonstrate that DAX-1 is expressed in Sertoli cells of rat testis. This expression is regulated during spermatogenesis and peaks during the androgen-sensitive phase of the spermatogenic cycle. In addition, we show that DAX-1 expression in Sertoli cells is regulated developmentally. Maximum levels are present in the rat between postnatal days 20 and 30, during the first spermatogenic wave. Moreover, we show that activation of the cAMP-signaling pathway by the pituitary hormone FSH leads to a potent down-regulation of DAX-1 expression in cultured Sertoli cells. This down-regulation requires transcription and de novo protein synthesis. Taken together, these data indicate that DAX-1 expression in Sertoli cells may influence the development of spermatogenic cells in response to steroid and pituitary hormones.


The ancestral role of ATP hydrolysis in type II topoisomerases: prevention of DNA double-strand breaks.
Type II DNA topoisomerases (topos) catalyse changes in DNA topology by passing one double-stranded DNA segment through another. This reaction is essential to processes such as replication and transcription, but carries with it the inherent danger of permanent double-strand break (DSB) formation. All type II topos hydrolyse ATP during their reactions; however, only DNA gyrase is able to harness the free energy of hydrolysis to drive DNA supercoiling, an energetically unfavourable process. A long-standing puzzle has been to understand why the majority of type II enzymes consume ATP to support reactions that do not require a net energy input. While certain type II topos are known to 'simplify' distributions of DNA topoisomers below thermodynamic equilibrium levels, the energy required for this process is very low, suggesting that this behaviour is not the principal reason for ATP hydrolysis. Instead, we propose that the energy of ATP hydrolysis is needed to control the separation of protein-protein interfaces and prevent the accidental formation of potentially mutagenic or cytotoxic DSBs. This interpretation has parallels with the actions of a variety of molecular machines that catalyse the conformational rearrangement of biological macromolecules.


Ru(II) Phenanthroline-Based Oligothienyl Complexes as Phototherapy Agents.
Ru(II) polypyridyl complexes have gained widespread attention as photosensitizers for photodynamic therapy (PDT). Herein, we systematically investigate a series of the type [Ru(phen)2(IP-nT)]2+, featuring 1,10-phenanthroline (phen) coligands and imidazo[4,5-f][1,10]phenanthroline ligands tethered to n = 0-4 thiophene rings (IP-nT). The complexes were characterized and investigated for their electrochemical, spectroscopic, and (photo)biological properties. The electrochemical oxidation of the nT unit shifted by -350 mV as n = 1 → 4 (+920 mV for Ru-1T, +570 mV for Ru-4T); nT reductions were observed in complexes Ru-3T (-2530 mV) and Ru-4T (-2300 mV). Singlet oxygen quantum yields ranged from 0.53 to 0.88, with Ru-3T and Ru-4T being equally efficient (∼0.88). Time-resolved absorption spectra of Ru-0T-1T were dominated by metal-to-ligand charge-transfer (3MLCT) states (τTA = 0.40-0.85 μs), but long-lived intraligand charge-transfer (3ILCT) states were observed in Ru-2T-4T (τTA = 25-148 μs). The 3ILCT energies of Ru-3T and Ru-4T were computed to be 1.6 and 1.4 eV, respectively. The phototherapeutic efficacy against melanoma cells (SK-MEL-28) under broad-band visible light (400-700 nm) increases as n = 0 → 4: Ru-0T was inactive up to 300 μM, Ru-1T-2T were moderately active (EC50 ∼ 600 nM, PI = 200), and Ru-3T (EC50 = 57 nM, PI > 1100) and Ru-4T (EC50 = 740 pM, PI = 114,000) were the most phototoxic. The activity diminishes with longer wavelengths of light and is completely suppressed for all complexes except Ru-3T and Ru-4T in hypoxia. Ru-4T is the more potent and robust PS in 1% O2 over seven biological replicates (avg EC50 = 1.3 μM, avg PI = 985). Ru-3T exhibited hypoxic activity in five of seven replicates, underscoring the need for biological replicates in compound evaluation. Singlet oxygen sensitization is likely responsible for phototoxic effects of the compounds in normoxia, but the presence of redox-active excited states may facilitate additional photoactive pathways for complexes with three or more thienyl groups. The 3ILCT state with its extended lifetime (30-40× longer than the 3MLCT state for Ru-3T and Ru-4T) implicates its predominant role in photocytotoxicity.


Analytic-domain lens design with proximate ray tracing.
We have developed an alternative approach to optical design which operates in the analytical domain so that an optical designer works directly with rays as analytical functions of system parameters rather than as discretely sampled polylines. This is made possible by a generalization of the proximate ray tracing technique which obtains the analytical dependence of the rays at the image surface (and ray path lengths at the exit pupil) on each system parameter. The resulting method provides an alternative direction from which to approach system optimization and supplies information which is not typically available to the system designer. In addition, we have further expanded the procedure to allow asymmetric systems and arbitrary order of approximation, and have illustrated the performance of the method through three lens design examples.


Retrospective evaluation of systemic hypertension in dogs with nonassociative (primary) immune-mediated hemolytic anemia (December 2016 to April 2019): 26 cases.
To report the prevalence of arterial hypertension in a population of dogs with nonassociative immune-mediated hemolytic anemia (IMHA) on presentation and during hospitalization. To determine the relationships of systolic blood pressure (SBP) with mortality and a prognostic indicator, the canine hemolytic anemia objective score. Retrospective observational study (December 2016 to April 2019). University teaching hospital. Twenty-six clinical dogs presenting to the ICU with nonassociative (primary) IMHA and a control group of 23 clinical dogs with idiopathic epilepsy hospitalized in the ICU for seizure treatment or monitoring. None. Hypertension was defined as SBP ≥ 160 mm Hg and severe hypertension as SBP ≥ 180 mm Hg. Mean SBP was significantly increased in IMHA dogs (161 mm Hg, SD = 21) compared to ICU control dogs (138 mm Hg, SD = 14; P < 0.005). Hypertension was present in 13 of 26 (50.0%) dogs across the period of hospitalization and was severe in three of 26 (11.5%). During at least 1 day of hospitalization, 18 of 26 (69.2%) dogs were hypertensive and eight of 26 (34.6%) were severely hypertensive. Hypertension was not associated with short-term mortality or canine hemolytic anemia objective score. In this retrospective study, hypertension was more prevalent in dogs with nonassociative IMHA than a control population of ICU-hospitalized dogs. An association between autoimmune conditions and hypertension has been previously reported in people but not within a canine population. Hypertension in dogs may have an inflammatory or autoimmune etiology. SBP should be monitored closely in canine IMHA, in case antihypertensive treatment is required.


Mechanisms of alcoholic liver injury.
There have been numerous recent advances in the understanding of the mechanisms of alcoholic liver disease pathogenesis. Endotoxin-induced Kupffer cell activation plays a role in cytokine-mediated inflammatory changes in the liver, and this can be blocked by a diet high in saturated fat, by a diet containing lactobacillus, which does not produce endotoxin, by neomycin antibiotic sterilization of the gut, by eliminating Kupffer cells, or by removing tumour necrosis factor-alpha with antibody or by using tumour necrosis factor-alpha knockout mice. The fatty liver component is mainly the result of the nicotinamide adenine dinucleotide/reduced nicotinamide adenine dinucleotide redox shift to the reduced state by ethanol oxidation generation of reduced nicotinamide adenine dinucleotide, although this too can be blocked by a diet high in saturated fat. Hepatocytic enlargement occurs due to ethanol-induced inhibition of the ubiquitin-proteasome pathway of cytoplasmic protein degradation and the retention of oxidized proteins in hepatocytes. The liver is scarred by stellate cells that have been activated by inflammatory cytokines and growth factors produced by activated Kupffer cells, and by bile ductule metaplasia. Mallory bodies and balloon cell degeneration develop through the ethanol-induced oxidative stress-protein kinase activation pathway, inhibition of phosphatase activity and inhibition of the ubiquitin-proteasome pathway.


The impact of baculovirus challenge on immunity: The effect of dose and time after infection.
Understanding how hosts respond to pathogen attack is crucial to disease management. The response of a host can be particularly important if hosts have to defend against multiple pathogens which could either benefit from or be suppressed by prior pathogen exposure. Insect defence against viruses is less well understood than responses to other entomopathogens and much of the information available relates to in vitro studies and model systems. Baculoviruses are natural pathogens of insects, particularly Lepidoptera, and have been well-studied in terms of their ecology, pest control potential and molecular biology. In order to examine how an insect reacts to baculovirus challenge, we measured components of the cellular and humoral immune response of the cabbage looper Trichoplusia ni to Trichoplusia ni SNPV, a narrow-host range nucleopolyhedrovirus (NPV), over four doses and three times after pathogen challenge (18, 42 and 90 h). We found that total haemocyte numbers peaked at 42 h post-exposure at all doses, and declined linearly with increasing dose after the 18 h time point. Two immune-related enzymes, phenoloxidase (PO) and FAD-glucose dehydrogenase (GLD), showed very different responses. PO levels were lowest at the 42 h time point and were not influenced by virus dose when each time point was examined separately. GLD levels declined over time but they interacted with virus dose in a non-linear manner, such that there was an increase in levels at intermediate virus doses after 18 h, no effect at 42 h, and then declined as infection progressed at 90 h post-infection. These data suggest that baculoviruses can rapidly infect haemocytes (or cause a reduction in their numbers) in a dose-dependent manner once the infection is systemic, likely reducing the ability of the host to counter subsequent infections. However, the data do not support a direct role for PO in defence against baculoviruses. Whether GLD plays a role in virus defence is still unclear.


Vasoactivity of rucaparib, a PARP-1 inhibitor, is a complex process that involves myosin light chain kinase, P2 receptors, and PARP itself.
Therapeutic inhibition of poly(ADP-ribose) polymerase (PARP), as monotherapy or to supplement the potencies of other agents, is a promising strategy in cancer treatment. We previously reported that the first PARP inhibitor to enter clinical trial, rucaparib (AG014699), induced vasodilation in vivo in xenografts, potentiating response to temozolomide. We now report that rucaparib inhibits the activity of the muscle contraction mediator myosin light chain kinase (MLCK) 10-fold more potently than its commercially available inhibitor ML-9. Moreover, rucaparib produces additive relaxation above the maximal degree achievable with ML-9, suggesting that MLCK inhibition is not solely responsible for dilation. Inhibition of nitric oxide synthesis using L-NMMA also failed to impact rucaparib's activity. Rucaparib contains the nicotinamide pharmacophore, suggesting it may inhibit other NAD+-dependent processes. NAD+ exerts P2 purinergic receptor-dependent inhibition of smooth muscle contraction. Indiscriminate blockade of the P2 purinergic receptors with suramin abrogated rucaparib-induced vasodilation in rat arterial tissue without affecting ML-9-evoked dilation, although the specific receptor subtypes responsible have not been unequivocally identified. Furthermore, dorsal window chamber and real time tumor vessel perfusion analyses in PARP-1-/- mice indicate a potential role for PARP in dilation of tumor-recruited vessels. Finally, rucaparib provoked relaxation in 70% of patient-derived tumor-associated vessels. These data provide tantalising evidence of the complexity of the mechanism underlying rucaparib-mediated vasodilation.


Occupational outcomes of people with multiple sclerosis during the COVID-19 pandemic: a systematic review with meta-analysis.
People with Multiple Sclerosis (PwMS) are vulnerable to unfavorable occupational outcomes and the COVID-19 pandemic brought major consequences on people's professional lives. In this view, we decided to investigate the occupational outcomes of PwMS during the COVID-19 pandemic. We performed a systematic review with meta-analysis searching key terms in four databases. We initially included any peer-reviewed original article that enrolled adult patients with the diagnosis of MS and assessed any occupational variable during the COVID-19 pandemic. There were no time limits and no language restrictions. The primary outcomes were the prevalence of unemployment, retirement and employment status change among people with MS during the COVID-19 pandemic. Other outcomes included the modality and characteristics of work: type of work, full-time work, part-time work and remote work. We also searched for data from studies that addressed any change in the work status due to the COVID-19 outbreak. We identified 49 eligible articles comprising a total sample size of 17,364 individuals with MS. The pooled prevalence of unemployment and retirement was 0.47 (95% CI = 0.42-0.53). The pooled prevalence of PwMS who were unemployed or retired was positively associated with the progressive phenotype of the disease (p = 0.017) and the use of glatiramer acetate (p = 0.004), but negatively associated with hospitalization due to COVID-19 (p = 0.008) and the use of immunosuppressants (p = 0.032), siponimod (p < 0.001), and cladribine (p = 0.021). The pooled proportion of PwMS that reported any change of the employment status during the COVID-19 pandemic was 0.43 (95% CI = 0.36-0.50) while the pooled prevalence of PwMS who worked remotely during this period was 0.37 (95% CI = 0.15-0.58). The change in employment status was negatively associated with the duration of MS (p = 0.03) but positively associated with the progressive phenotype of the disease (p < 0.001). Our seminal review may serve as an example of how patients with neurological diseases or disabilities in general may have their jobs impacted in a pandemic and foster the context of global socio-economic crisis.


Clinicopathological features and prognoses in younger and older patients with gastric cancer.
Patients of different ages with gastric cancer (GC) have different clinicopathological features and prognoses. The results for different crowds are limited and controversial. The aim of this study was to investigate the differences in clinicopathological features and prognoses between younger and older GC patients. From January 2007 to December 2011, a consecutive total of 112 GC patients under 41 years old and 358 GC patients over 69 years old who underwent gastrectomy for GC were recruited for this study. Then, the clinicopathological features and prognoses of these patients were analyzed comparatively. The gender, differentiation, carbohydrate antigen (CA) 19-9 and carcinoembryonic antigen (CEA) were significantly different between younger and older GC patients. There were more female and undifferentiated younger GC patients, and there were higher percentages of positive CA19-9 and CEA in older GC patients. The number of metastatic lymph nodes was an independent risk parameter for prognosis in younger patients, and the AJCC TNM (Tumor-Nodes-Metastases classification by American Joint Committee on Cancer) stage, radicality and tumor size were independent risk parameters for prognosis in older GC patients. Younger GC patients have a much better prognoses with lower monocyte-to-lymphocyte ratio and higher prognostic nutritional index than older patients. Younger GC patients have better immunity and nutritional status and better prognoses. The number of metastatic lymph nodes was the only risk parameter for prognosis in younger GC patients. We should take more effective treatments for younger GC patients with lymph nodes metastasis and pay more attention to the nutritional problems of older GC patients.


Reliability of fine-needle aspiration in patients with familial nonmedullary thyroid cancer.
In this case-control study we describe how often thyroid cancers and occult cancers are diagnosed or not diagnosed by fine-needle aspiration (FNA) in patients with thyroid nodules and a family history of nonmedullary thyroid cancers (FNMTC). Our hypothesis is that patients with thyroid nodules and a family history of FNMTC seem to be similar to patients with thyroid nodules and a history of exposure to low-dose therapeutic radiation. Both have been reported to have multifocal thyroid neoplasms and malignant tumors are common. Cytological examination may therefore be less accurate. From 1979 to 1996, 27 patients from 24 families with FNMTC were examined histologically after a preoperative cytological examination in all of them. A positive cytology examination was defined when biopsy documented thyroid cancer. It was interpreted as a false-negative study when a benign diagnosis was made and thyroid cancer was present anywhere within the thyroid, including in areas sampled or not sampled by FNA and not palpable preoperatively. A randomized control group, matched for age and gender, contained 27 patients with papillary thyroid cancer without familial disease. In our study group, 25 patients were treated with total thyroidectomy, including 7 with neck dissection, and 2 by thyroid lobectomy. At final histological examination 17 of 27 patients (63%) in this study group had multiple nodules and 25 of 27 (92.6%) had thyroid cancer. Thyroid cancer was diagnosed by FNA in 22 of 25 patients (88%), with 3 (12%) false-negative biopsies due to sampling errors (thyroid cancer not in the index nodule), versus 1 (3.7%) false-negative biopsy in the control group. Two patients in the study group with benign nodules were accurately diagnosed. In patients with false-negative biopsies and a history of FNMTC, the cancer was situated in one or more small nodules. Only one cancer was occult (< 1.0 cm). One-third of the patients in our study group (33%) had a history of radiation; 44% of the irradiated group had a single nodule; 56% had multiple nodules. In the control group, 9 of 27 patients (33%) also had a history of radiation; 33% of the irradiated group had a single nodule, 67% had multiple nodules. In conclusion, the reliability of FNA in patients with FNMTC appears to be less accurate than it is for other patients because of the high incidence of multifocal thyroid cancer and coexistence of benign nodules. Patients with thyroid nodules and a family history of thyroid cancer are more likely to have thyroid cancer and because they also have more coexistent benign nodules, they must be followed closely or treated with total or near-total thyroidectomy.


A novel rotate-and-fire digital spiking neuron and its neuron-like bifurcations and responses.
A novel rotate-and-fire digital spiking neuron is presented. The digital neuron is a wired system of shift registers and thus it is suited to on-chip learning unlike many other analog spiking neuron models. By adjusting the wiring pattern among the registers, the digital neuron can generate spike trains with various spike patterns and can exhibit related bifurcations. A discrete-continuous hybrid map, which describes the neuron dynamics without any approximation, is derived analytically. Using the hybrid map, it is shown that the digital spiking neuron can mimic typical bifurcation phenomena and various nonlinear responses of biological neurons.


Resonance tube phonation in water: High-speed imaging, electroglottographic and oral pressure observations of vocal fold vibrations--a pilot study.
Phonation into glass tubes ('resonance tubes'), keeping the free end of the tube in water, has been a frequently used voice therapy method in Finland and more recently also in other countries. The purpose of this exploratory study was to investigate what effects tube phonation with and without water has on the larynx. Two participants were included in the study. The methods used were high-speed imaging, electroglottographic observations of vocal fold vibrations, and measurements of oral pressure during tube phonation. Results showed that the fluctuation in the back pressure during tube phonation in water altered the vocal fold vibrations. In the high-speed imaging, effects were found in the open quotient and amplitude variation of the glottal opening. The open quotient increased with increasing water depth (from 2 cm to 6 cm). A modulation effect by the water bubbles on the vocal fold vibrations was seen both in the high-speed glottal area tracings and in the electroglottography signal. A second experiment revealed that the increased average oral pressure was largely determined by the water depth. The increased open quotient can possibly be explained by an increased abduction of the vocal folds and/or a reduced transglottal pressure. The back pressure of the bubbles also modulates glottal vibrations with a possible 'massage' effect on the vocal folds. This effect and the well-defined average pressure increase due to the known water depth are different from those of other methods using a semi-occluded vocal tract.


Ultrafast x-ray photoelectron spectroscopy in the microsecond time domain.
We introduce a new approach for ultrafast in situ high-resolution X-ray photoelectron spectroscopy (XPS) to study surface processes and reaction kinetics on the microsecond timescale. The main idea is to follow the intensity at a fixed binding energy using a commercial 7 channeltron electron analyzer with a modified signal processing setup. This concept allows for flexible switching between measuring conventional XP spectra and ultrafast XPS. The experimental modifications are described in detail. As an example, we present measurements for the adsorption and desorption of CO on Pt(111), performed at the synchrotron radiation facility BESSY II, with a time resolution of 500 μs. Due to the ultrafast measurements, we are able to follow adsorption and desorption in situ at pressures of 2 × 10(-6) mbar and temperatures up to 500 K. The data are consistently analyzed using a simple model in line with data obtained with conventional fast XPS at temperatures below 460 K. Technically, our new approach allows measurement on even shorter timescales, down to 20 μs.


Can telemonitoring reduce hospitalization and cost of care? A health plan's experience in managing patients with heart failure.
Telemonitoring provides a potentially useful tool for disease and case management of those patients who are likely to benefit from frequent and regular monitoring by health care providers. Since 2008, Geisinger Health Plan (GHP) has implemented a telemonitoring program that specifically targets those members with heart failure. This study assesses the impact of this telemonitoring program by examining claims data of those GHP Medicare Advantage plan members who were enrolled in the program, measuring its impact in terms of all-cause hospital admission rates, readmission rates, and total cost of care. The results indicate significant reductions in probability of all-cause admission (odds ratio [OR] 0.77; P<0.01), 30-day and 90-day readmission (OR 0.56, 0.62; P<0.05), and cost of care (11.3%; P<0.05). The estimated return on investment was 3.3. These findings imply that telemonitoring can be an effective add-on tool for managing elderly patients with heart failure.


Genomewide linkage scan for bipolar-disorder susceptibility loci among Ashkenazi Jewish families.
The relatively short history of linkage studies in bipolar disorders (BPs) has produced inconsistent findings. Implicated regions have been large, with reduced levels of significance and modest effect sizes. Both phenotypic and genetic heterogeneity may have contributed to the failure to define risk loci. BP is part of a spectrum of apparently familial affective disorders, which have been organized by severity. Heterogeneity may arise because of insufficient data to define the spectrum boundaries, and, in general, the less-severe disorders are more difficult to diagnose reliably. To address the inherent complexities in detecting BP susceptibility loci, we have used restricted diagnostic classifications and a genetically more homogeneous (Ashkenazi Jewish) family collection to perform a 9-cM autosomal genomewide linkage scan. Although they are genetically more homogeneous, there are no data to suggest that the rate of illness in the Ashkenazim differs from that in other populations. In a genome scan of 41 Ashkenazi pedigrees with a proband affected with bipolar I disorder (BPI) and at least one other member affected with BPI or bipolar II disorder (BPII), we identified four regions suggestive of linkage on chromosomes 1, 3, 11, and 18. Follow-up genotyping showed that the regions on chromosomes 1, 3, and 18 are also suggestive of linkage in a subset of pedigrees limited to relative pairs affected with BPI. Furthermore, our chromosome 18q22 signal (D18S541 and D18S477) overlaps with previous BP findings. This research is being conducted in parallel with our companion study of schizophrenia, in which, by use of an identical approach, we recently reported significant evidence for a schizophrenia susceptibility locus in the Ashkenazim on chromosome 10q22.


Educational effort and CQI program improves ordering of serum digoxin levels.
In a 289-bed, acute-care, non-tertiary-care regional referral center, a continuous quality improvement educational effort successfully improved ordering of serum digoxin levels. Following a determination that physicians were not requesting serum digoxin levels correctly, the P & T Committee issued a letter describing appropriate wording of orders for serum digoxin levels. The number of incorrectly ordered serum digoxin levels requested dropped significantly over a 49-month period as a result of the intervention described below, which also resulted in cost savings.


An endemic frog harbors multiple expression loci with different patterns of variation in the MHC class II B gene.
The Chinese concave-eared torrent frog (Odorrana tormota) was the first non-mammalian vertebrate demonstrated to both produce and perceive ultrasonic frequencies. Due to habitat degradation caused by human activity, currently, the species is at the edge of extinction. In this study, we demonstrated that the species harbors at least four expressed major histocompatibility complex (MHC) class II B loci. Four distinct intron 2 sequences and three intron 1 sequences were isolated from genomic DNA. Additionally, eight alleles were recovered from three cDNA samples, of which five could be assigned to three individual loci while the locus identity of the remaining three alleles could not be determined. Based on the intron sequences, locus-specific primers were designed for Odto-A, Odto-B, and Odto-C. Of these loci, Odto-B was monomorphic in Huangshan population (n = 32), and Odto-C displayed low allelic polymorphism with only three similar sequences obtained from partial individuals. In contrast, Odto-A showed high allelic diversity with 20 unique alleles recovered from the population. No inter- or intra-allele recombination was detected. However, 10 positive selected amino acid residues and an excess of non-synonymous substitutions were detected in the putative antigen binding sites (ABS), these results combined with trans-species evolution suggested some form of balancing selection that maintained higher allelic polymorphism for the locus. Our findings demonstrate that despite the effects of a severely shrinking population, O. tormota exhibits high polymorphism compared to other amphibians, both in terms of the allele number and diversity across the MHC class II B loci.


[The agglutination test of coracidia of pseudophyllidean cestodes and its use in cestodological research].
Ability of coracidia to agglutination in homologous antiserum was established. The method of reaction of a direct agglutination of coracidia (RDAC) in the culture of alive larvae was described. When comparing the RDAC results in homologous and heterologous variants a dependence of the agglutination pattern on the degree of antiserum homology was established. On the example of RDAC of the cestodes Diphyllobothrium dendriticum and D. ditremum in polyvalent and species-specific antisera to D. dendriticum possibility was shown of RDAC usage in the studies on specific taxonomy of pseudophyllid cestodes.


Challenges with using primer IDs to improve accuracy of next generation sequencing.
Next generation sequencing technologies, like ultra-deep pyrosequencing (UDPS), allows detailed investigation of complex populations, like RNA viruses, but its utility is limited by errors introduced during sample preparation and sequencing. By tagging each individual cDNA molecule with barcodes, referred to as Primer IDs, before PCR and sequencing these errors could theoretically be removed. Here we evaluated the Primer ID methodology on 257,846 UDPS reads generated from a HIV-1 SG3Δenv plasmid clone and plasma samples from three HIV-infected patients. The Primer ID consisted of 11 randomized nucleotides, 4,194,304 combinations, in the primer for cDNA synthesis that introduced a unique sequence tag into each cDNA molecule. Consensus template sequences were constructed for reads with Primer IDs that were observed three or more times. Despite high numbers of input template molecules, the number of consensus template sequences was low. With 10,000 input molecules for the clone as few as 97 consensus template sequences were obtained due to highly skewed frequency of resampling. Furthermore, the number of sequenced templates was overestimated due to PCR errors in the Primer IDs. Finally, some consensus template sequences were erroneous due to hotspots for UDPS errors. The Primer ID methodology has the potential to provide highly accurate deep sequencing. However, it is important to be aware that there are remaining challenges with the methodology. In particular it is important to find ways to obtain a more even frequency of resampling of template molecules as well as to identify and remove artefactual consensus template sequences that have been generated by PCR errors in the Primer IDs.


A role of p75 in NGF-mediated down-regulation of the A(2A) adenosine receptors in PC12 cells.
Nerve growth factor (NGF) induces differentiation of the rat pheochromocytoma clone (PC12) by activating the high affinity receptor, p140(trkA), linked to mitogen-activated protein kinase. While the physiological role of the low affinity NGF receptor (p75) has not been clearly defined, this receptor promotes activation of nuclear factor (NF) kappaB in Schwann cells. PC12 cells express the A(2A) adenosine receptor (AR), whose expression is significantly decreased by NGF treatment. In this study, we determined whether TrkA or p75 is involved in NGF-mediated regulation of A(2A)AR expression. NGF treatment decreased A(2A)AR in a time-dependent manner, with maximal effects observed by 1 day, and continued down-regulation of the receptor for up to 3 days in the presence of NGF. The decrease in A(2A)AR was associated with a more delayed decrease in the steady-state levels of the A(2A)AR mRNA. Down-regulation of the A(2A)AR at 1 day was mimicked by activators of NFkappaB, such as H(2)O(2), and ceramide, and was attenuated by the inhibitor pyrrolidine dithiocarbamate or following transient transfection of PC12 cells with a dominant negative IkappaBalpha mutant. Moreover, NGF stimulated nuclear accumulation of p65 subunits of NFkappaB (but not p50 subunits) in PC12 cells, as determined by electrophoretic mobility shift assays and by Western blotting. In contrast, inhibition of TrkA by AG879 or of TrkA-dependent mitogen-activated protein kinase mitogen-activated protein kinase kinase with PD98059 blocked PC12 cell differentiation without affecting A(2A)AR down-regulation, suggesting dissociation between these two phenomena. Taken together, these data provide strong support for the involvement of the p75/NFkappaB pathway in NGF-mediated down-regulation of A(2A)AR in PC12 cells.


Pu-erh Tea Extract Attenuates Nicotine-Induced Foam Cell Formation in Primary Cultured Monocytes: An in Vitro Mechanistic Study.
In this study, the mechanisms by which pu-erh tea extract (PETE) attenuates nicotine-induced foam cell formation were investigated. Monocytes were purified from healthy individuals using commercial antibodies coated with magnetic beads. We found that the nicotine-induced (1-10 μM) expression of oxidized low-density lipoprotein receptors (ox-LDLRs) and α9-nAchRs in monocytes was significantly attenuated by 24 h of PETE (10 μg/mL; ∗, p < 0.05) cotreatment. Nicotine (1 μM for 24 h) significantly induced the expression of the surface adhesion molecule ICAM-1 and the monocyte integrin adhesion molecule (CD11b) by human umbilical vein endothelial cells (HUVECs) and triggered monocytes to differentiate into macrophages via interactions with the endothelium. After treatment with nicotine (0.1-10 μM for 24 h), the HUVECs released chemotactic factors (IL-8) to attract monocytes into the tunica intima of the artery, and the monocytes then transformed into foam cells. We demonstrated that PETE treatment (>1 μg/mL for 24 h; ∗, p < 0.05) significantly attenuates nicotine-induced (1 μM) monocyte migration toward HUVECs and foam cell formation. This study suggests that tea components effectively attenuate the initial step (foam cell formation) of nicotine-induced atherosclerosis in circulating monocytes.


Endoscopic evaluation of the navicular bursa: observations, treatment and outcome in 92 cases with identified pathology.
Diagnostic navicular bursoscopy has been described in limited cases. Review of greater numbers is needed to define its contribution to case management and prognostic values. To report: 1) clinical, diagnostic and endoscopic findings in a series of cases, 2) surgical techniques and case outcomes and 3) prognostic values. The authors hypothesise: 1) lameness localising to the navicular bursa is commonly associated with dorsal border deep digital flexor tendon (DDFT) lesions, 2) endoscopy allows extent of injuries to be assessed and treated, 3) case outcome relates to severity of DDFT injury and 4) the technique is safe and associated with little morbidity. All horses that underwent endoscopy of a forelimb navicular bursa for investigation of lameness were identified. Case files were reviewed and those with injuries within the bursa selected for further analysis. One-hundred-and-fourteen horses were identified. Ninety-two had injuries within the bursa and DDFT injuries were identified in 98% of bursae. Of those examined with magnetic resonance imaging (MRI), 56% had combination injuries involving the DDFT and navicular bone. Sixty-one percent of horses returned to work sound, 42% returned to previous performance. Horses with extensive tearing and combination injuries of the DDFT and navicular bone identified with MRI, had worse outcomes. Lameness localising to the navicular bursa is commonly associated with injuries to the dorsal border of the DDFT. Endoscopy permits identification and characterisation of injuries within the navicular bursa and enables lesion management. Outcome following debridement is related to severity of injury but overall is reasonable. Horses with lameness localising to the navicular bursa may have tears of the DDFT. Bursoscopy is able to contribute diagnostic and prognostic information and debridement of lesions improves outcome compared to cases managed conservatively.


[Clinical value of the cholesterol test in patients with arteriosclerosis].
Capacity for the excretion of cholesterol (CS) and bile acids was studied, using an original cholesterol load test, in atherosclerotic patients. They showed reduced fecal excretion of neutral sterols and bile acids following cholesterol load, unlike normal subjects, whose respective values were increased irrespective of age. Fecal excretion of these substances was particularly low in second-type hyperlipoproteinemia. Atherosclerosis-related changes of sterol balance may be an evidence of latent incompetence of mechanisms of hepatic evacuation of substances that may be regarded as atherosclerotic risk factors.


Spatial dynamics of porcine epidemic diarrhea (PED) spread in the southern Kyushu, Japan.
Porcine epidemic diarrhea (PED) was detected for the first time in seven years in Japan in October 2013 in Okinawa Prefecture. By December 2013, PED had spread into Miyazaki and Kagoshima Prefectures in the southern part of Kyushu, one of the regions with the highest farm density in the country. The objective of the study here was to assess the spatial dynamics of PED spread during the 8 months of the epidemic in the southern part of Kyushu between December 2013, the month observed first case in the studied region, and July 2014. Information on location and capacity of all farms in the prefectures (n=1269) was obtained from a government database containing demographic information for livestock producers. Additionally, data on PED detection (positive or negative) was obtained from the regional Livestock Hygiene Service Center. The Cuzick-Edwards (CE) test, the Knox test, the directional test, and the permutation model of the scan statistic were used to assess the spatio-temporal distribution of the epidemic. PED cumulative farm level incidence was 19.5% (248/1269) through the study period. The highest density of positive farms was observed in the most farm-populated areas of the prefecture. The CE test revealed an extensive degree of spatial clustering, with clustering of positive sites being significant (P<0.01) up to the 35th level of neighborhood (approximately 5km in the studied data). The observed-to-expected ratio of cases was maximized at short spatio-temporal distances, with values of the observed-to-expected ratio of cases maximized when the thresholds were set at 2km and 10days, respectively. A significant (P<0.01) direction of spread was detected towards the northeastern direction. The permutation model detected five significant (P<0.01) clusters occurring at different stages of the epidemic wave. The strong spatio-temporal clustering of PED-infected farms during the first 6 months of the epidemic in the southern part of Kyushu is consistent with results obtained elsewhere and demonstrates the rapid spread of the virus in naïve populations.


Genetic diversity in farm animals--a review.
Domestication of livestock species and a long history of migrations, selection and adaptation have created an enormous variety of breeds. Conservation of these genetic resources relies on demographic characterization, recording of production environments and effective data management. In addition, molecular genetic studies allow a comparison of genetic diversity within and across breeds and a reconstruction of the history of breeds and ancestral populations. This has been summarized for cattle, yak, water buffalo, sheep, goats, camelids, pigs, horses, and chickens. Further progress is expected to benefit from advances in molecular technology.


Fetal human neural progenitors can be the target for tumor transformation.
The hypothesis that stem cells may seed cancer has emerged from the cancer stem cells concept. However, the experimental systems necessary to provide more direct evidence to support the hypothesis have been lacking. We have used fetal neural progenitor cells (hNPC) transduced with the telomerase hTERT gene to investigate the neoplastic potential of hNPCs. The hTERT-transduced line, hNPCs-G3 lost normal diploid karyotype, showed loss of contact inhibition, anchorage independence, and formed neuroblastoma-like tumours in all of 10 mice. These data suggest that hNPCs have the potential for neoplastic transformation. These data have implications for providing a novel tool to test the feasibility of new anticancer treatment strategies and raise the possibility of a risk for the use of hNPCs in cell transplantation.


Love and hooking up in the new millennium: communication technology and relationships among urban African American and Puerto Rican young adults.
Communication technology is a central feature of young people's lives, but its role in romantic and sexual relationships has not been thoroughly examined. This article describes how young adults use communication technology for partnering across relationship stages (formation, maintenance, and dissolution) and types (serious/casual), and proposes implications of usage in relationships. This study analyzed qualitative data from a five-week, prospective, coital diary method with related debriefing interviews (N = 70) of African American and Puerto Rican men and women aged 18 to 25 years in Hartford and Philadelphia. Cell phones, including calls, text messaging, and mobile Internet, were the most common forms of communication technology used for partnering goals. Participants reported using cell phones to pursue partnering goals across all relationship stages, including formation (meeting, screening, and getting to know new partners), maintaining existing relationships, and breaking up. Cell phone uses depended on the type of relationship (serious/casual) and the participants' intentions and desires. Results indicated that cell phones are an important element of communication among young adults in romantic and sexual relationships. Specific features of cell phone communication shape the process and context of partnering. Future research should explore emerging communication technologies and implications for psychosocial development, dating violence, and sexual behavior.


Herpes simplex virus esophagitis in the immunocompetent host: an overview.
The aim of this study was to delineate the characteristics of herpes simplex virus esophagitis (HSVE) in the immunocompetent host. The study entailed a case report and a review of relevant literature through a MEDLINE search back to 1966. All cases with documented HSVE in patients without immunosuppression were selected and their characteristics defined. A total of 38 cases were identified. The age range was 1-76 yr and the male/female ratio 3.2/1. Antecedent exposure to HSV disease was described in eight cases (21.1%). A prodrome of systemic manifestations preceded the onset of esophageal symptoms in nine subjects (23.6%). Manifestations included acute odynophagia (76.3%), heartburn (50%), and fever (44.7%). Concurrent oropharyngeal lesions were uncommon (n = 8, 21.1%). Endoscopically, extensive involvement was common, showing friable mucosa (84.2%), numerous ulcers (86.8%), and whitish-exudates (39.5%). The distal esophagus was most commonly affected (63.8%). Microscopic examination showed characteristic viral cytopathology in 26 (68.4%) cases. Virus was recovered from esophageal-brushes or biopsies in 23 of 24 (95.8%) patients and immunocytochemistry was positive in seven of eight (87.5%) cases. Immune status was consistent with primary HSV infection in eight (21.1%) cases. The disease was self-limiting, although esophageal perforation and upper GI bleeding were reported in one case each. HSVE in the immunocompetent host is a rare but distinct entity, and is significantly more common in male subjects. It represents either primary infection or reactivation, and is characterized by acute onset, systemic manifestations, and extensive erosive-ulcerative involvement of the mid-distal esophagus. Histopathological examination alone may miss the diagnosis; adding tissue-viral culture optimizes the diagnostic sensitivity. It is usually self-limiting; whether antiviral therapy is beneficial remains unknown.


Studies in vivo and in vitro on the uptake and degradation of soluble collagen alpha 1(I) chains in rat liver endothelial and Kupffer cells.
Intravenously administered 125I-labelled monomeric alpha 1 chains (125I-alpha 1) of collagen type I were rapidly cleared and degraded by the liver of rats. Isolation of the liver cells after injection of the label revealed that the uptake per liver endothelial cell equalled the uptake per Kupffer cell, whereas the amount taken up per hepatocyte was negligible. The uptake of 125I-alpha 1 in cultured cells was 10 times higher per liver endothelial cell than per Kupffer cell. The ligand was efficiently degraded by cultures of both cell types. However, spent medium from cultures of Kupffer cells, unlike that from cultures of other cells, contained gelatinolytic activity which degraded 125I-alpha 1. The presence of hyaluronic acid, chondroitin sulphate or mannose/N-acetylglucosamine-terminal glycoproteins, which are endocytosed by the liver endothelial cells via specific receptors, did not interfere with binding, uptake or degradation of 125I-alpha 1 by these cells. Unlabelled alpha 1 and heat-denatured collagen inhibited the binding to a much greater extent than did native collagen. The presence of fibronectin or F(ab')2 fragments of anti-fibronectin antibodies did not affect the interaction of the liver endothelial cells, or of other types of liver cells, with 125I-alpha 1. The accumulation of fluorescein-labelled heat-denatured collagen in vesicles of cultured liver endothelial cells is evidence that the protein is internalized. Moreover, chloroquine, 5-dimethylaminonaphthalene-1-sulphonylcadaverine (dansylcadaverine), monensin and cytochalasin B, which impede one or more steps of the endocytic process, inhibited the uptake of 125I-alpha 1 by the liver endothelial cells. Leupeptin, an inhibitor of cathepsin B and 'collagenolytic cathepsins', inhibited the intralysosomal degradation of 125I-alpha 1, but had no effect on the rate of uptake of the ligand. The current data are interpreted as follows. (1) The ability of the liver endothelial cells and the Kupffer cells to sequester circulating 125I-alpha 1 efficiently may indicate a physiological pathway for the breakdown of connective-tissue collagen. (2) The liver endothelial cells express receptors that specifically recognize and mediate the endocytosis of collagen alpha 1(I) monomers. (3) The receptors also recognize denatured collagen (gelatin). (4) Fibronectin is not involved in the binding of alpha 1 to the receptors. (5) Degradation occurs intralysosomally by leupeptin-inhibitable cathepsins.


Deoxynucleotide-interconverting enzymes and the quantification of deoxynucleoside triphosphates in mammalian cells.
We have demonstrated that methanol extracts of human cells are heterogeneous with regard to content of dNDP (deoxynucleoside diphosphate) and dNMP (deoxynucleoside monophosphate) kinases. The presence of these enzymes can affect the reliability of techniques used to measure intracellular pools of deoxynucleotides. An optimized extraction procedure and enzymic assay for dNTP species in haematopoietic cells are described which provide sensitivity to measure 0.1-40pmol of dATP, dTTP and dGTP, and 1.0-40pmol of dCTP. The extraction and assay give linear results with (2.5-15)x10(6) nucleated cells and (0.1-1.5)x10(9) red blood cells. Under these conditions, extracts equivalent to ~0.5x10(6) nucleated haematopoietic cells catalyse the phosphorylation of 0-8% of dNDP and dNMP standards to dNTP and incorporate them into deoxynucleotide polymer under circumstances where 100% of an equimolar dNTP standard would be incorporated. By contrast, extracts of 0.4x10(6) HeLa cells totally converted dADP, dTDP and dGDP into dNTP with subsequent polymerization. Conversion of dCDP was somewhat less efficient. The results demonstrate conclusively that the activities of deoxynucleotide interconverting enzymes differ in different types of human cells. They can interfere with assay of nucleotides, but may not do so in many types of cell extracts. In particular, dNTP concentrations can be measured in human haematopoietic cells after extraction with 60% (v/v) methanol and are not artificially elevated by deoxynucleotide interconversions. It is apparent that extraction and assay procedures for measurement of dNTP species should be analysed for each cell type in order to minimize contaminating enzyme activities and ensure accuracy of dNTP quantification.


Comparative study on gingival retraction using mechanochemical procedure and pulsed Nd = YAG laser irradiation.
The aim of the present study was to study the tissue reaction and gingival healing in pulsed laser gingival retraction in comparison with chemico-mechanical gingival retraction technique. This work was applied on six patients recommended for orthodontic extraction of first premolars upper and lower. The patients were divided into three groups (two patients each). -One group with laser tissue retraction. -Another group with ferric sulphate (13.3%). -Third group with aluminium chloride (25%). The present histologic findings revealed that with the application of pulsed Nd: YAG laser the gingival tissues showed faster healing with less hemorrhage and less inflammatory reaction in comparison with the other two groups. In conclusion it was evident that pulsed laser is a surgical device increasingly important to dentistry. The present study can support with the clinical application of laser in gingival retraction as a simple convenient, painless method.


Risk factors for emergency department revisit in elderly patients with gastrointestinal bleeding secondary to anticoagulant therapy.
To evaluate the frequency of emergency department (ED) revisits among elderly patients with gastrointestinal bleeding secondary to anticoagulant treatment and identify factors associated with an increased risk of ED revisits. A 3-year retrospective observational study was designed, including elderly patients (≥65 years) with atrial fibrillation and undergoing oral anticoagulation therapy who visited the ED for gastrointestinal bleeding. To evaluate the risk factors for 30-day revisit, a multivariate analysis was designed including comorbidities, concomitant treatment, change in anticoagulant treatment and prescription of direct-acting oral anticoagulants. 80 patients were included. At discharge, anticoagulation therapy was modified in 21 (26.2%) patients; and changed from an oral anticoagulant to heparin in 17 (21.2%) patients and to another oral anticoagulant in 4 (5.0%) patients. Anticoagulant treatment was withdrawn in 5 (6.3%) patients at discharge. Eleven (13.7%) patients revisited the ED 30 days after hospital discharge for bleeding episodes. No differences in the frequency of revisit to the ED were observed in the patients who changed their anticoagulant treatment at discharge. In the multivariate analysis, chronic kidney disease was the only factor significantly associated with revisits at 30 days. Elderly patients who experience a first episode of gastrointestinal bleeding have a high risk of revisiting the ED for a bleeding episode, with no particular differences between the types of anticoagulant prescribed at discharge.


Periodontitis, coronary heart disease and myocardial infarction: treat one, benefit all.
: Periodontal disease is conventionally defined as an inflammatory condition affecting the tissues surrounding and supporting the teeth (i.e. gum and periodontium). Recent statistics show that the prevalence of this condition is continuously growing worldwide, thus raising severe healthcare concerns, not only for local problems emerging from poor oral health, but also for the potential risk of developing systemic complications. Therefore, this article aims to provide an update on the intriguing association between periodontitis, coronary heart disease (CHD) and/or myocardial infarction (MI). Taken together, the available published information seems to support the existence of a significant association between periodontitis and CHD, whilst the risk of acute ischemic cardiac events appears magnified in patients with preexisting coronary artery disease. This epidemiological link is supported by reliable biological evidence, showing that periodontal disease may unfavourably modulate the cardiovascular risk, whereby patients with periodontitis have increased frequency of overweight, hypertension, endothelial dysfunction, dyslipidaemia, platelet hyper-reactivity, and may also be characterized by a prothrombotic state. Apart from these critical atherogenic factors, translocation of periodontal microorganisms into the bloodstream, and their further accumulation within atherosclerotic plaques, would contribute to enhance plaque instability and the risk of developing acute ischemic coronary events. Interesting evidence is also emerging that local or systemic statins administration could be beneficial for safeguarding periodontal health, thus enlightening the intriguing relationship existing between CHD and periodontitis.


Cationic Porphyrin-Based Ionic Nanomedicines for Improved Photodynamic Therapy.
This study presents the synthesis and characterization of monosubstituted cationic porphyrin as a photodynamic therapeutic agent. Cationic porphyrin was converted into ionic materials by using a single-step ion exchange reaction. The small iodide counteranion was replaced with bulky BETI and IR783 anions to reduce aggregation and enhance the photodynamic effect of porphyrin. Carrier-free ionic nanomedicines were then prepared by using the reprecipitation method. The photophysical characterization of parent porphyrin, ionic materials, and ionic nanomaterials, including absorbance, fluorescence and phosphorescence emission, quantum yield, radiative and nonradiative rate, and lifetimes, was performed. The results revealed that the counteranion significantly affects the photophysical properties of porphyrin. The ionic nanomaterials exhibited an increase in the reactive oxygen yield and enhanced cytotoxicity toward the MCF-7 cancer cell line. Examination of results revealed that the ionic materials exhibited an enhanced photodynamic therapeutic activity with a low IC50 value (nanomolar) in cancerous cells. These nanomedicines were mainly localized in the mitochondria. The improved light cytotoxicity is attributed to the enhanced photophysical properties and positive surface charge of the ionic nanomedicines that facilitate efficient cellular uptake. These results demonstrate that ionic material-based nanodrugs are promising photosensitizers for photodynamic therapy.


A pilot study on Ayurvedic management of oral submucous fibrosis.
Oral submucous fibrosis (OSMF) is a chronic debilitating disease and a well-recognized potentially premalignant condition of the oral cavity. Various medical and surgical treatment modalities have been used in modern science, but results are not satisfactory owing to recurrence, adverse effects, and sometimes worsening the condition. On analyzing the disease condition with Ayurvedic approach, it seems to be nearer to Vata-Pitta dominant chronic Sarvasara Mukharoga and needs to be treated at local as well as systemic level. To evaluate the effect of proposed Ayurvedic treatment protocol in the patients of OSMF. It was an open-label nonrandomized clinical trial with black box design comprising of holistic Ayurvedic approach in which 22 patients of OSMF completed the treatment. In all of them after Koshthashuddhi (mild purgation) and Shodhana Nasya (errhine therapy); Pratisarana (external application) with Madhupippalyadi Yoga, Kavala (gargling) with Ksheerabala Taila and internally Rasayana Yoga were given for 2 months and followed for 1 month. It revealed statistically highly significant relief in almost all signs and symptoms as well in inter incisal distance improvement. Furthermore, sustained relief was found in follow-up. Ayurvedic treatment protocol is effective in the management of OSMF.


Identification of valid reference genes for the normalization of RT qPCR gene expression data in human brain tissue.
Studies of gene expression in post mortem human brain can contribute to understanding of the pathophysiology of neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Quantitative real-time PCR (RT qPCR) is often used to analyse gene expression. The validity of results obtained using RT qPCR is reliant on accurate data normalization. Reference genes are generally used to normalize RT qPCR data. Given that expression of some commonly used reference genes is altered in certain conditions, this study aimed to establish which reference genes were stably expressed in post mortem brain tissue from individuals with AD, PD or DLB. The present study investigated the expression stability of 8 candidate reference genes, (ubiquitin C [UBC], tyrosine-3-monooxygenase [YWHAZ], RNA polymerase II polypeptide [RP II], hydroxymethylbilane synthase [HMBS], TATA box binding protein [TBP], beta-2-microglobulin [B2M], glyceraldehyde-3-phosphate dehydrogenase [GAPDH], and succinate dehydrogenase complex-subunit A, [SDHA]) in cerebellum and medial temporal gyrus of 6 AD, 6 PD, 6 DLB subjects, along with 5 matched controls using RT qPCR (TaqMan(R) Gene Expression Assays). Gene expression stability was analysed using geNorm to rank the candidate genes in order of decreasing stability in each disease group. The optimal number of genes recommended for accurate data normalization in each disease state was determined by pairwise variation analysis. This study identified validated sets of mRNAs which would be appropriate for the normalization of RT qPCR data when studying gene expression in brain tissue of AD, PD, DLB and control subjects.


[Study of dorsal erection osteotomy in kyphosis].
Presented is a procedure for the correction of the juvenile kyphosis and the kyphotic false posture upon spondylitis ankylopoietica. Upon spine dissects the extraction and outbreak force of screws and screw pairs dorsally brought into the vertebral body were measured. It has been showed that the pedicle screws tolerate very high extraction forces upon sagittal and right-angle stress as well. The correction of the juvenile kyphosis upon rigid kyphosis and the correction by lordosing the thoraco-lumbar transition upon patients with a spondylitis ankylopoietica is clinically possible with good results.


Molecular Hybridization-Inspired Optimization of Diarylbenzopyrimidines as HIV-1 Nonnucleoside Reverse Transcriptase Inhibitors with Improved Activity against K103N and E138K Mutants and Pharmacokinetic Profiles.
Molecular hybridization is a powerful strategy in drug discovery. A series of novel diarylbenzopyrimidine (DABP) analogues were developed by the hybridization of FDA-approved drugs etravirine (ETR) and efavirenz (EFV) as potential HIV-1 nonnucleoside reverse transcriptase inhibitors (NNRTIs). Substituent modifications resulted in the identification of new DABPs with the combination of the strengths of the two drugs, especially compound 12d, which showed promising activity toward the EFV-resistant K103N mutant. 12d also had a favorable pharmacokinetic (PK) profile with liver microsome clearances of 14.4 μL/min/mg (human) and 33.2 μL/min/mg (rat) and an oral bioavailability of 15.5% in rat. However, its activity against the E138K mutant was still unsatisfactory; E138K is the most prevalent NNRTI resistance-associated mutant in ETR treatment. Further optimizations resulted in a highly potent compound (12z) with no substituents on the phenyl ring and a 2-methyl-6-nitro substitution pattern on the 4-cyanovinyl-2,6-disubstitued phenyl motif. The antiviral activity of this compound was much higher than those of ETR and EFV against the WT, E138K, and K103N variants (EC50 = 3.4, 4.3, and 3.6 nM, respectively), and the cytotoxicity was decreased while the selectivity index (SI) was increased. In particular, this compound exhibited acceptable intrinsic liver microsome stability (human, 34.5 μL/min/mg; rat, 33.2 μL/min/mg) and maintained the good PK profile of its parent compound EFV and showed an oral bioavailability of 16.5% in rat. Molecular docking and structure-activity relationship (SAR) analysis provided further insights into the binding of the DABPs with HIV-1 reverse transcriptase and provided a deeper understanding of the key structural features responsible for their interactions.


Construction of a model cell line for the assay of MDR1 (multi drug resistance gene-1) substrates/inhibitors using HeLa cells.
Cancer cells often become resistant to chemotherapy, and induction of the ABC transporter Multi-drug Resistance gene-1 (MDR1) is a major cause. We established a tool for high-throughput screening of substrates and inhibitors of MDR1, using transformed HeLa cells that over-express MDR1. The cDNA for human MDR1 was subcloned into the eukaryotic expression vector pBK-CMV to produce an MDR1 expression vector, pBK-CMV/MDR1. HeLa cells were transfected with pBK-CMV/MDR1 or the empty vector pBK-CMV. Transfection of the vector sequence for MDR1 and its expression were evaluated by genomic PCR and western blotting, respectively. The efficiency of the MDR1 transporter for pumping a substrate out of the transformed cells was evaluated using rhodamine123 (R-123), a mitochondrial dye that is also an MDR1 substrate. After treatment of the MDR1-expressing HeLa cells with MDR1 substrate vinblastin or inhibitors cyclosporin A and verapamil, the amount of R-123 retained in the cells was increased to 2 to 2.3 times the level in untreated MDR1-expressing HeLa cells. The transfection of empty pBK-CMV had no effect on the R-123 retention in HeLa cells, regardless of drug treatment. In conclusion, we have established a model human carcinoma cell line that expresses functional MDR1 and can be used to screen for substrates and inhibitors of MDR1.


Patients With Cancer Searching for Cancer- or Health-Specific Web-Based Information: Performance Test Analysis.
Searching the internet for cancer-related information helps patients with cancer satisfy their unmet information needs and empowers them to play a more active role in the management of their disease. However, to benefit from the search, patients need a sufficient level of skill to search, select, appraise, and apply web-based health information. We aim to study the operational, navigational, information, and evaluation skills and problems of patients with cancer performing cancer-related search tasks using the internet. A total of 21 patients with cancer were recruited during their stay at the rehabilitation clinic for oncological rehabilitation. Participants performed eight cancer-related search tasks using the internet. The participants were asked to think aloud while performing the tasks, and the screen activities were recorded. The types and frequencies of performance problems were identified and coded into categories following an inductive coding process. In addition, the performance and strategic characteristics of task execution were summarized descriptively. All participants experienced problems or difficulties in executing the tasks, and a substantial percentage of tasks (57/142, 40.1%) could not be completed successfully. The participants' performance problems were coded into four categories, namely operating the computer and web browser, navigating and orientating, using search strategies, and evaluating the relevance and reliability of web-based information. The most frequent problems occurred in the third and fourth categories. A total of 90% (19/21) of participants used nontask-related search terms or nonspecific search terms. A total of 95% (20/21) of participants did not control for the source or topicality of the information found. In addition, none of the participants verified the information on 1 website with that on another website for each task. A substantial group of patients with cancer did not have the necessary skills to benefit from cancer-related internet searches. Future interventions are needed to support patients in the development of sufficient internet-searching skills, focusing particularly on information and evaluation skills.


Prevalence and risk factors for diabetic microvascular complications in newly diagnosed type II diabetes mellitus. Sankara Nethralaya Diabetic Retinopathy Epidemiology and Molecular Genetic Study (SN-DREAMS, report 27).
The aims of this study were to report the prevalence of various microvascular complications and to identify the various clinical and biochemical characteristics related to these complications in subjects with newly diagnosed type II diabetes. Of the 5999 subjects enumerated, 1414 subjects with diabetes (both known and newly diagnosed) were analyzed for the study. Among the diabetic subjects, 248 (17.5%) were newly diagnosed with diabetes and the remaining had history of diabetes. All subjects underwent a detailed standard evaluation to detect diabetic retinopathy (fundus photography), neuropathy (vibration pressure threshold), and nephropathy (microalbuminuria). The prevalence of any form of microvascular complication was 30.2% (95% confidence interval [CI]=24.5-35.9). The prevalence of diabetic retinopathy was 4.8%, and that of diabetic nephropathy and neuropathy was 10.5%. The risk factors for developing any form of microvascular complication were increasing age (odds ratio [OR]=1.07, 95% CI=1.04-1.11, P<.0001), increasing systolic blood pressure (OR=1.03, 95% CI=1.01-1.06, P=.001), and increasing hemoglobin (OR=1.39, 95% CI=1.09-1.79, P=.011). The risk factors for diabetic retinopathy and diabetic nephropathy were increasing systolic blood pressure (OR=1.06 [P=.001] for retinopathy and OR=1.04 [P=.012] for nephropathy) and increasing hemoglobin (OR=2.20 [P=.007] for retinopathy and OR=1.57 [P=.023] for nephropathy). The risk factor for diabetic neuropathy was increasing age (OR=1.12, P<.0001). Nearly one third of the newly diagnosed type II diabetes subjects had some form of microvascular complication; nephropathy, and neuropathy being commoner than retinopathy.


Screening and modulation of extracellular signals by mucous barrier. Serum glycosylphosphatidylinositol phospholipase D (GPI-PLD) releases protective mucous barrier from oral mucosa.
Performance of mucosal epithelial barrier is modified by numerous agents that exert effects on mucin- Mucin Binding Protein (MBP) complex. The aim ofthe studies described was to determine the nature of the damage or modification of oral mucous barrier by the short-term exposure to ethanol. Culture of rat buccal mucosa in the presence of ethanol and [3H]-labeled proline and palmitate revealed substantial decrease in MBP synthesis and the release of MBPto the medium. The radioscanning of the samples prepared from the culture medium and the apical epithelial membranes subjected to SDS-PAGE and western blotting disclosed that the released, water soluble 97kDa MBP glycopeptide was labeled with proline and palmitate. When the experiments were conducted in the presence of 5mM EDTA, the GPI-PLD inhibitor, the majority of radiolabeled MBP remained in the membrane-bound form and was extractable with Triton X- 114. The results on the purified GPI-linked MBP degradation by serum enzyme, by the saliva containing serum transudate, and the suppression of the process by inclusion of GPI-PLD-specific inhibitor support our contention that membrane MBP is released to medium by GPI-PLD-like activity. The release of MBP from apical epithelial surfaces was induced by depletion of mucin and the presence of serum-derived GPI-PLD in the tissue homogenate. Strong likelihood exists that under in situ conditions ethanol-induced transudation of serum to saliva provides the vehicle for the transfer of GPI-PLD activity to salivary contents. Defacement of the oral surfaces from mucous barrier signals prospect of lumenal agent influence on the unprotected epithelial exterior, and allows ingression of microbes and untoward acting substances into the organism.


Selective sigma-1 receptor antagonists for the treatment of pain.
The sigma-1 receptor (σ1R) is located in areas of the CNS key for pain control and belongs to a unique target class with chaperoning functions over different molecular targets involved in transmission and amplification of nociceptive messages. Preclinical evidence supports a role for σ1R antagonists in the treatment of pain states where hypersensitivity develops as hyperalgesia and allodynia, two common symptoms encountered in neuropathic and other chronic pain conditions. Additionally, σ1R antagonists increase opioid analgesia without increasing opioid-related unwanted effects, which point to their potential use as opioid adjuvant therapy. This review summarizes the structure and function of the σ1R as well as the medicinal chemistry and pharmacological studies directed to the identification of σ1R antagonists for the treatment of pain.


Leaky cell model of hard spheres.
We study packings of hard spheres on lattices. The partition function, and therefore the pressure, may be written solely in terms of the accessible free volume, i.e., the volume of space that a sphere can explore without touching another sphere. We compute these free volumes using a leaky cell model, in which the accessible space accounts for the possibility that spheres may escape from the local cage of lattice neighbors. We describe how elementary geometry may be used to calculate the free volume exactly for this leaky cell model in two- and three-dimensional lattice packings and compare the results to the well-known Carnahan-Starling and Percus-Yevick liquid models. We provide formulas for the free volumes of various lattices and use the common tangent construction to identify several phase transitions between them in the leaky cell regime, indicating the possibility of coexistence in crystalline materials.


More than a "touch of the flu": a response to Mayrhuber et al's ""with fever it's the real flu I would say": laypersons' perception of common cold and influenza and their differences - a qualitative study in Austria, Belgium and Croatia".
This short reply contests two assumptions made by the authors of Mayrhuber et al's. "With fever it's the real flu I would say." The first is that there is influenza can be reliably defined by a medical case definition. The second is that this small qualitative study can be generalisable. However, it does underline the important point that technical diagnostic terms may be used on different registers by a variety of actors in the medical setting.


Stereotactic radiation therapy in oligometastatic colorectal cancer: outcome of 102 patients and 150 lesions.
To evaluate the local control (LC), progression free survival (PFS), out-field PFS, overall survival (OS), toxicity and failure predictors of SRT in a series of various sites oligometastatic CRC patients. Patients with oligometastatic CRC disease were analyzed retrospectively. The SRT prescribed dose was dependent on the lesion volume and its location. 102 consecutive oligometastatic CRC patients (150 lesions) were included. They underwent SRT between 2012 and 2015. Median prescription dose was 45 Gy (median dose/fraction was 15 Gy/3 fractions biological equivalent dose (BED10) 112.5 Gy). Median follow-up was 11.4 months. No patients experienced G3 and G4 toxicity. No progression was found in 82% (radiological response at 3 months) and 85% (best radiological response) out of 150 evaluable lesions. At 1 and 2 years: LC was 70% and 55%; OS was 90% and 90%; PFS was 37% and 27%; out-field PFS was 37% and 23% respectively. Progressive disease was correlated with BED10 (better LC when BED10 was ≥ 75 Gy (p < 0.0001)). In multivariate analysis, LC was higher in lesions with a Plpnning target volume (PTV) volume < 42 cm3 and BED10 ≥ 75 Gy. Patients with Karnofsky performance status < 90 showed higher out-field progression. SRT is an effective treatment for patients with oligometastases from CRC. Its low treatment-associated morbidity and acceptable LC make of SRT an option not only in selected cases. Further studies should be focused to clarify which patient subgroup will benefit most from this treatment modality and to define the optimal dose to improve LC while maintaining low toxicity profile.


Potential role of bile acids in the pathogenesis of necrotizing enterocolitis.
Necrotizing enterocolitis (NEC) is one of the most common acute gastrointestinal diseases in preterm infants. Recent studies have found that NEC is not only caused by changes in the intestinal environment but also by the failure of multiple systems and organs, including the liver. The accumulation of bile acids (BAs) in the ileum and the disorder of ileal BA transporters are related to the ileum injury of NEC. Inflammatory factors such as tumor necrosis factor (TNF)-α and interleukin (IL)-18 secreted by NEC also play an important role in regulating intrahepatic BA transporters. As an important link connecting the liver and intestinal circulation, the bile acid metabolic pathway plays an important role in the regulation of intestinal microbiota, cell proliferation, and barrier protection. In this review, we focus on how bile acids explore the dynamic changes of bile acid metabolism in necrotizing enterocolitis and the potential therapeutic value of targeting the bile acid signaling pathways.


Escherichia coli phnN, encoding ribose 1,5-bisphosphokinase activity (phosphoribosyl diphosphate forming): dual role in phosphonate degradation and NAD biosynthesis pathways.
An enzymatic pathway for synthesis of 5-phospho-D-ribosyl alpha-1-diphosphate (PRPP) without the participation of PRPP synthase was analyzed in Escherichia coli. This pathway was revealed by selection for suppression of the NAD requirement of strains with a deletion of the prs gene, the gene encoding PRPP synthase (B. Hove-Jensen, J. Bacteriol. 178:714-722, 1996). The new pathway requires three enzymes: phosphopentomutase, ribose 1-phosphokinase, and ribose 1,5-bisphosphokinase. The latter activity is encoded by phnN; the product of this gene is required for phosphonate degradation, but its enzymatic activity has not been determined previously. The reaction sequence is ribose 5-phosphate --> ribose 1-phosphate --> ribose 1,5-bisphosphate --> PRPP. Alternatively, the synthesis of ribose 1-phosphate in the first step, catalyzed by phosphopentomutase, can proceed via phosphorolysis of a nucleoside, as follows: guanosine + P(i) --> guanine + ribose 1-phosphate. The ribose 1,5-bisphosphokinase-catalyzed phosphorylation of ribose 1,5-bisphosphate is a novel reaction and represents the first assignment of a specific chemical reaction to a polypeptide required for cleavage of a carbon-phosphorus (C-P) bond by a C-P lyase. The phnN gene was manipulated in vitro to encode a variant of ribose 1,5-bisphosphokinase with a tail consisting of six histidine residues at the carboxy-terminal end. PhnN was purified almost to homogeneity and characterized. The enzyme accepted ATP but not GTP as a phosphoryl donor, and it used ribose 1,5-bisphosphate but not ribose, ribose 1-phosphate, or ribose 5-phosphate as a phosphoryl acceptor. The identity of the reaction product as PRPP was confirmed by coupling the ribose 1,5-bisphosphokinase activity to the activity of xanthine phosphoribosyltransferase in the presence of xanthine, which resulted in the formation of 5'-XMP, and by cochromatography of the reaction product with authentic PRPP.


[Non-drug rehabilitation of patients with chronic obstructive pulmonary disease concurrent with hypertension].
to evaluate the efficiency of decimeter wave therapy and halotherapy, which were additionally added to basic therapy, in patients with chronic obstructive pulmonary disease (COPD) concurrent with hypertension at the inpatient stage. 36 patients aged 20 to 75 years with Stages I-II COPD concurrent with Stages I-II, first-second grade hypertension were examined and treated. The clinical examination included collection of complaints and medical history data, clinical laboratory and instrumental (electrocardiography, spirography) studies, and health-related quality of life (using the SF-36 questionnaire). The patients were randomized into two groups: a study group and a comparison group. The study group patients received decimeter wave therapy and halotherapy in addition to basic drug treatment; the comparison patients had basic drug therapy. Pre- and postoperative comparative analysis of the major clinical manifestations of comorbidities revealed more pronounced positive changes with the lower rate of clinical manifestations in the study group. It was also observed to have a more marked reduction in blood pressure (BP) with its goal levels achieved. The mean pulse BP decreased by 28% in the study group (p=0.000005) and did not statistically reduced in the comparison group. In the study group patients, the integral quality-of-life indicator after a package of medical rehabilitation measures became statistically significantly higher by 35%. This indicator in the comparison group was statistically significantly unchanged. The directionality of the proposed rehabilitation complex towards the common pathogenetic components of the development and progression of COPD and hypertension, as well as the high efficiency of the complex justify its appropriate inclusion in the combination treatment and rehabilitation of this category of patients.


Some studies on the composition and surface properties of oil bodies from the seed cotyledons of safflower (Carthamus tinctorius) and linseed (Linum ustatissimum).
1. The average oil-body diameter in intact cells of developing linseed (Linum usitatissimum) and safflower (Carthamus tinctorius) cotyledons was similar (about 1.4 micrometer), and there was little change in size after oil bodies were isolated and repeatedly washed. 2. The glycerolipid composition of washed oil bodies from both developing and mature cotyledons of the two species was similar; oil bodies from ten different batches of cotyledons contained 4.3 +/- 0.16 mumol of 3-sn-phosphatidylcholine and 25.2 +/- 1.7 mumol of diacylglycerol per 1000 mumol of triacylglycerol. During four successive washings of a once-washed oil-body preparation, the proportion of diacylglycerol to triacylglycerol remained constant and that of 3-sn-phosphatidylcholine to triacylglycerol decreased by only 20%. 3. The protein content of thrice-washed oil bodies from the two species was similar, about 2.4% of the weight of glycerolipids, and appeared to be independent of the stage of cotyledon maturity. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated that the protein of purified oil bodies from the two species consisted mainly of only four polypeptides and that two of the polypeptides from each species had apparent mol.wts. of 17500 and 15500. Similar patterns of polypeptides were obtained after the hydrolysis of the 15500-mol.wt. polypeptides from linseed and safflower oil bodies by Staphylococcus aureus V8 proteinase, whereas the proteolysis of the 17500-mol.wt. polypeptides from the two species produced different patterns of polypeptides. 4. The 3-sn-phosphatidylcholine in oil-body preparations was hydrolysed about 85% by bee-venom phospholipase A2 without any apparent coalescence of the oil bodies. Incubation with lipase from Rhizopus arrhizus caused rapid coalescence of the oil bodies, and this lipase appeared to initially hydrolyse diacylglycerols in preference to triacylglycerol. 5. Oil bodies from both species were almost completely dispersed in suspensions of pH between 7.1 and 8.3, but formed large aggregates at pH values between 6.7 and 3.9; pH-induced aggregation caused no coalescence. Aggregates formed under acidic conditions were dispersed by re-adjusting the pH of suspensions to 8.3. 6. A freeze-etch electron-microscopic examination of isolated oil bodies indicated that these organelles were bounded by some form of membrane with a particle-free outer surface.


Charlson score is a robust predictor of 30-day complications following spinal metastasis surgery.
Retrospective chart review. To identify predictors of 30-day complications after the surgical treatment of spinal metastasis. Surgical treatment of spinal metastasis is considered palliative with the aim of reducing or delaying neurologic deficit. Postoperative complication rates as high as 39% have been reported in the literature. Complications may impact patient quality of life and increase costs; therefore, an understanding of which preoperative variables best predict 30-day complications will help risk-stratify patients and guide therapeutic decision making and informed consent. We retrospectively reviewed 200 cases of spinal metastasis surgically treated at Stanford Hospital between 1999 and 2009. Multiple logistic regression was performed to determine which preoperative variables were independent predictors of 30-day complications. Sixty-eight patients (34%) experienced one or more complications within 30 days of surgery. The most common complications were respiratory failure, venous thromboembolism, and pneumonia. On multivariate analysis, Charlson Comorbidity Index score was the most significant predictor of 30-day complications. Patients with a Charlson score of two or greater had over five times the odds of a 30-day complication as patients with a score of zero or one. After adjusting for demographic, oncologic, neurologic, operative, and health factors, Charlson score was the most robust predictor of 30-day complications. A Charlson score of two or greater should be considered a surgical risk factor for 30-day complications, and should be used to risk-stratify surgical candidates. If complications are anticipated, medical staff can prepare in advance, for instance, scheduling aggressive ICU care to monitor for and treat complications. Finally, Charlson score should be controlled for in future spinal metastasis outcomes studies and compared to other comorbidity assessment tools.


Synthesis and photoluminescence properties of silica-modified SiO2@ANA-Si-Tb@SiO2, SiO2@ANA-Si-Tb-L@SiO2 core-shell-shell nanostructured composites.
Three novel core-shell nanostructured composites SiO2@ANA-Si-Tb, SiO2@ANA-Si-Tb-L (L = second ligand) with SiO2 as the core and terbium organic complex as the shell were successfully synthesized. The core and shell were connected together by covalent bonds. The terbium ion was coordinated with organic ligand-forming terbium organic complex in the shell layer. The organosilane (HOOCC5H4NN(CONH(CH2)3Si(OCH2CH3)3)2 (abbreviated as ANA-Si) was used as the first ligand and 1,10-phenanthroline (phen) or 2-thenoyltrifluoroacetone (TTA) was used as the second ligand. Furthermore, silica-modified SiO2@ANA-Si-Tb@SiO2, SiO2@ANA-Si-Tb-L@SiO2 core-shell-shell nanostructured composites were also synthesized by sol-gel chemical route, which involved the hydrolysis and polycondensation processes of tetraethoxysilane (TEOS) using cetyltrimethyl ammonium bromide (CTAB) as a surface-active agent. An amorphous silica shell was coated around the SiO2@ANA-Si-Tb, SiO2@ANA-Si-Tb-L core-shell nanostructured composites. The core-shell and core-shell-shell nanostructured composites exhibited excellent luminescence in the solid state. Meanwhile, an improved luminescent stability property of the core-shell-shell nanostructured composites was observed for the aqueous solution. This type of core-shell-shell nanostructured composites exhibited bright luminescence, high stability and good solubility, which may present potential applications in the fields of optoelectronic devices, bio-imaging, medical diagnosis and study on the structure of function composite materials.


Speeding up the evaluation of new agents in cancer.
Despite both the increase in basic biologic knowledge and the fact that many new agents have reached various stages of development during the last 10 years, the number of new treatments that have been approved for patients has not increased as expected. We propose the multi-arm, multi-stage trial design as a way to evaluate treatments faster and more efficiently than current standard trial designs. By using intermediate outcomes and testing a number of new agents (and combinations) simultaneously, the new design requires fewer patients. Three trials using this methodology are presented.


Dysfunctional neural networks associated with impaired social interactions in early psychosis: an ICA analysis.
The "default mode", or baseline of brain function is a topic of great interest in schizophrenia research. Recent neuroimaging studies report that the symptoms of chronic schizophrenia subjects are associated with temporal frequency alterations as well as with the disruption of local spatial patterns in the default mode network (DMN). Previous studies both on chronic and medicated subjects with psychosis suffered from limitations; on this basis, it was hypothesized that the default mode network showed abnormal activation and connectivity in young and neuroleptic-naïve patients with first-episode psychosis. This study investigated emotional responses to pleasant and unpleasant/disgusting visual stimuli by a resting-state analysis of fMRI-data from 12 untreated first-episode psychosis patients with prevalently negative symptomatology versus 12 healthy subjects. We chose this experimental task to explore the functional link between default mode network and hedonic processing which has been proposed as a marker of cerebral dysfunction in psychotic disorder and implicated in its pathophysiology. Independent Component Analysis (ICA) was used to identify the default mode component. Both healthy and first-episode subjects showed significant spatial differences in the default mode network. In first-episode subjects, medial frontal hypoactivity and cerebellar hyperactivity were correlated with the severity of negative symptoms.


Is Mpox a Sexually Transmitted Infection? Why Narrowing the Scope of This Disease May Be Harmful.
The 2022 multinational mpox outbreak has been characterized by unprecedented spread among men who have sex with men outside of sub-Saharan Africa. Close contact during sex and intimacy has been well established as a key pathway for human-to-human transmission in the current outbreak. Discussions on whether to assign this illness as a sexually transmitted infection (STI) have been ongoing since the initiation of the outbreak. While sexual contact certainly appears to be a primary means of spread, classifying mpox as an STI is inaccurate based on its known transmission dynamics, yields potential unintended consequences, and ignores the historical impact of the disease in Central and West Africa. Rather than focusing our energy on disease categorization, more effort should be placed on destigmatizing this illness and empowering communities at risk to protect themselves from mpox.


Treatment of refractory non-Hodgkin's lymphoma with radiolabeled MB-1 (anti-CD37) antibody.
The biodistribution, toxicity, and therapeutic potential of anti-CD37 monoclonal antibody (MoAb) MB-1 labeled with iodine 131 (131I) was evaluated in ten patients with advanced-, low- or intermediate-grade non-Hodgkin's lymphomas who failed conventional treatment. Sequential dosimetric studies were performed with escalating amounts of antibody MB-1 (0.5, 2.5, 10 mg/kg) trace-labeled with 5 to 10 mCi 131I. Serial tumor biopsies and gamma camera imaging showed that the 10 mg/kg MoAb dose yielded the best MoAb biodistribution in the ten patients studied. Biodistribution studies in the five patients with splenomegaly and tumor burdens greater than 1 kg indicated that not all tumor sites would receive more radiation than normal organs, and these patients were therefore not treated with high-dose radioimmunotherapy. The other five patients did not have splenomegaly and had tumor burdens less than 0.5 kg; all five patients in this group showed preferential localization and retention of MoAb at tumor sites. Four of these patients have been treated with 131I (232 to 608 mCi) conjugated to anti-CD37 MoAb MB-1, delivering 850 to 4,260 Gy to tumor sites. Each of these four patients attained a complete tumor remission (lasting 4, 6, 11+, and 8+ months). A fifth patient, whose tumor did not express the CD37 antigen, was treated with 131I-labeled anti-CD20 MoAb 1F5 and achieved a partial response. Myelosuppression occurred 3 to 5 weeks after treatment in all cases, but there were no other significant acute toxicities. Normal B cells were transiently depleted from the bloodstream, but immunoglobulin (Ig) levels were not affected, and no serious infections occurred. Two patients required reinfusion of previously stored autologous, purged bone marrow. Two patients developed asymptomatic hypothyroidism 1 year after therapy. The tolerable toxicity and encouraging efficacy warrant further dose escalation in this phase I trial.


The Trigeminocardiac Reflex During the Anterior Transpetrosal Approach.
The anterior transpetrosal approach (ATPA), during which the trigeminal nerve (TN) is manipulated, has a risk of eliciting the trigeminocardiac reflex (TCR). The aim of this study was to assess the risk of TCR during ATPA. Surgical records of 92 patients who had surgical treatment via ATPA at the Keio University Hospital between December 2005 and June 2015 were retrospectively analyzed. Patients were divided into 2 groups on the basis of the occurrence of TCR during surgery, and clinical and tumor characteristics were compared. Tumor characteristics were evaluated based on preoperative images and intraoperative findings and included the side of the lesion, size of the lesion, tumor size in Meckel cave, pathology of the disease, cavernous sinus invasion, extension into Meckel cave, and adhesion of lesion to TN. TCR was observed in 14 of 92 patients (15.2%). TCR occurrence was significantly related to Meckel cave tumor size (P = 0.0264) and adhesion of the lesion to TN (P = 0.0002). This study suggests that TCR is related to Meckel cave tumor size and tumor adhesion to TN in ATPA. To our knowledge, this is the first report describing TCR during ATPA.


Molecular phylogeny and genome size evolution of the genus Betula (Betulaceae).
Betula L. (birch) is a genus of approx. 60 species, subspecies or varieties with a wide distribution in the northern hemisphere, of ecological and economic importance. A new classification of Betula has recently been proposed based on morphological characters. This classification differs somewhat from previously published molecular phylogenies, which may be due to factors such as convergent evolution, hybridization, incomplete taxon sampling or misidentification of samples. While chromosome counts have been made for many species, few have had their genome size measured. The aim of this study is to produce a new phylogenetic and genome size analysis of the genus. Internal transcribed spacer (ITS) regions of nuclear ribosomal DNA were sequenced for 76 Betula samples verified by taxonomic experts, representing approx. 60 taxa, of which approx. 24 taxa have not been included in previous phylogenetic analyses. A further 49 samples from other collections were also sequenced, and 108 ITS sequences were downloaded from GenBank. Phylogenetic trees were built for these sequences. The genome sizes of 103 accessions representing nearly all described species were estimated using flow cytometry. As expected for a gene tree of a genus where hybridization and allopolyploidy occur, the ITS tree shows clustering, but not resolved monophyly, for the morphological subgenera recently proposed. Most sections show some clustering, but species of the dwarf section Apterocaryon are unusually scattered. Betula corylifolia (subgenus Nipponobetula) unexpectedly clusters with species of subgenus Aspera Unexpected placements are also found for B. maximowicziana, B. bomiensis, B. nigra and B. grossa Biogeographical disjunctions were found within Betula between Europe and North America, and also disjunctions between North-east and South-west Asia. The 2C-values for Betula ranged from 0·88 to 5·33 pg, and polyploids are scattered widely throughout the ITS phylogeny. Species with large genomes tend to have narrow ranges. Betula grossa may have formed via allopolyploidization between parents in subgenus Betula and subgenus Aspera. Betula bomiensis may also be a wide allopolyploid. Betula corylifolia may be a parental species of allopolyploids in the subsection Chinenses Placements of B. maximowicziana, B. michauxii and B. nigra need further investigation. This analysis, in line with previous studies, suggests that section Apterocaryon is not monophyletic and thus dwarfism has evolved repeatedly in different lineages of Betula Polyploidization has occurred many times independently in the evolution of Betula.


Evaluation of fungal and yeast diversity in Slovakian wine-related microbial communities.
Since the yeast flora of Slovakian enology has not previously been investigated by culture-independent methods, this approach was applied to two most common cultivars Frankovka (red wine) and Veltlin (white wine), and complemented by cultivation. Model samples included grapes, initial must, middle fermenting must and must in the end-fermentation phase. The cultured isolates were characterized by length polymorphism of rDNA spacer two region using fluorescence PCR and capillary electrophoresis (f-ITS PCR), and some were identified by sequencing. The microbial DNA extracted directly from the samples without cultivation was analysed by f-ITS PCR, amplicons were cloned and sequenced. The use of universal fungal primers led to detection of both yeasts and filamentous fungi. The amplicon of highest intensity and present in all the samples corresponded to Hanseniaspora uvarum. Other species demonstrated by both approaches included Saccharomyces sp., Metschnikowia pulcherrima or M. chrysoperlae, Candida zemplinina, Cladosporium cladosporioides, Botryotinia fuckeliana, Pichia anomala, Candida railenensis, Cryptococcus magnus, Metschnikowia viticola or Candida kofuensis, Pichia kluyveri or Pichia fermentas, Pichia membranifaciens, Aureobasidium pullulans, Alternaria alternata, Erysiphe necator, Rhodotorula glutinis, Issatchenkia terricola and Debaryomyces hansenii. Endemism of Slovakian enological yeasts was suggested on the level of minor genetic variations of the known species and probably not accounting for novel species. The prevalence of H. uvarum over Saccharomyces sp. in the samples was indicated. This is the first culture-independent study of Slovakian enology and the first time f-ITS PCR profiling was used on wine-related microbial communities.


Effects of leukotrienes and f-Met-Leu-Phe on oxidative metabolism of neutrophils and eosinophils.
We assessed the effects of several leukotrienes and of f-Met-Leu-Phe on oxygen consumption in neutrophils and on the initial burst of chemiluminescence (CL) in both neutrophils and eosinophils. It was found that f-Met-Leu-Phe initiated 2.6 times higher oxygen consumption in neutrophils than did leukotriene B4 (LTB4). f-Met-Leu-Phe also stimulated five to 10 times more CL from both types of granulocytes than LTB4, which was at least five times more potent than its omega-hydroxylated metabolite, 20-OH-LTB4, whereas the corresponding 20-COOH derivative was effective only in eosinophils. The double dioxygenation product 5(S), 12(S)- DHETE caused no CL. Neutrophils from patients with chronic granulomatous disease did not respond with CL to any of the agents. The peak of CL occurred 50 to 60 sec after the addition of fMLP, whereas the LTB4-associated peak occurred after 5 to 6 sec and then rapidly subsided. The treatment of cells with sodium azide to inhibit the myeloperoxidase system did not change the kinetics or the rapid decline of the LTB4-induced CL. The CL response to LTB4 could be inhibited to 85% by 0.5 microgram/ml of superoxide dismutase, to 72% by 200 mg/ml of catalase, and to 50% by 80 microM of mannitol. The corresponding figures for f-Met-Leu-Phe-induced CL were 80, 58, and 16%, suggesting that, although a substantial part of the CL appears to be due to superoxide ion production, other oxygen radicals are involved in luminol-enhanced CL production. Thus, in contrast to some previous reports that leukotrienes do not stimulate an oxidative metabolic response in granulocytes despite their potent activity as chemotactic factors, our studies show that leukotrienes are definite inducers of granulocyte oxidative metabolism.


Anatomy of the clitoris and the female sexual response.
The clitoris may be the most pivotal structure for female sexual pleasure. While its significance has been reported for hundreds of years, no complete anatomical description was available until recently. Most of the components of the clitoris are buried under the skin and connective tissues of the vulva. It comprises an external glans and hood, and an internal body, root, crura, and bulbs; its overall size is 9-11 cm. Clitoral somatic innervation is via the dorsal nerve of the clitoris, a branch of the pudendal nerve, while other neuronal networks within the structure are complex. The clitoris is the center for orgasmic response and is embryologically homologous to the male penis. While the source of vaginal eroticism might or might not be exclusively clitoral stimulation, it is necessary to understand the intricate anatomy of the organ to assess the data in this regard. Ultimately, sexual enjoyment entails a balance of physical and emotional factors and should be encouraged.