Tick-Virus Interactions: Toll Sensing.
Ticks are important vectors of viruses that infect and cause disease in man, livestock, and companion animals. The major focus of investigation of tick-borne viruses has been the interaction with the mammalian host, particularly the mechanisms underlying disease and the development of vaccines to prevent infection. Only recently has research begun to investigate the interaction of the virus with the tick host. This is striking when considering that the virus spends far more time infecting the tick vector relative to the vertebrate host. The assumption has been that the tick host and virus have evolved to reach an equilibrium whereby virus infection does not impede the tick life cycle and conversely, the tick does not restrict virus replication and through blood-feeding on vertebrates, disseminates the virus. The development and application of new technologies to tick-pathogen interactions has been fuelled by a number of developments in recent years. This includes the release of the first draft of a tick genome, that of Ixodes scapularis, and the availability of tick-cell lines as convenient models to investigate interactions. One of the by-products of these investigations has been the observation of familiar proteins in new situations. One such protein family is Toll and Toll-like receptors that in vertebrates play a key role in detection of microorganisms, including viruses. But does Toll signaling play a similar role in detection of virus infection in ticks, and if it does, how does this affect the maintenance of viruses within the tick?


What is moving in silica at 1 K? A computer study of the low-temperature anomalies.
Though the existence of two-level systems (TLS) is widely accepted to explain low-temperature anomalies in many physical observables, knowledge about their properties is very rare. For silica, which is one of the prototype glass-forming systems, we elucidate the properties of the TLS via computer simulations by applying a systematic search algorithm. We get specific information in the configuration space, i.e., about relevant energy scales, the absolute number of TLS, and electric dipole moments. Furthermore, important insight about the real-space realization of the TLS can be obtained. Comparison with experimental observations is included.


[Guidelines on urolithiasis: update of diagnosis and treatment].
The Japanese urolithiasis clinical guideline was established in 2002. Since that time, the American Urological Associstion (AUA) and European Association of Urology (EAU) nephrolithiasis guideline panel has developed several guidelines on the management of urolithiasis. Recently, the joint AUA/EAU panel published a 2007 update of the original 1997 report on the management of ureteral calculi. The EAU panel yields the publication of guidelines on urolithiasis, with updates almost annually. These guidelines provide useful recommendations on the management of urolithiasis. Based on their findings, the most significant update of guideline concluded that both extracorporeal shockwave lithotripsy and ureteroscopy (URS) are acceptable first-line treatments for patients requiring ureteral stone removal. With the development of smaller caliber ureteroscopes and the introduction of improved instrumentation, including the holmium : yttrium aluminum garnet laser, URS has evolved into a safer and more efficacious modality for treatment of stones in all locations in the ureter during the last decade. The other change is the establishment of effective medical expulsive therapy to facilitate spontaneous stone passage.


Ongoing Clinical Trials of Vaccines to Fight against COVID-19 Pandemic.
Coronavirus disease 2019 (COVID-19) has developed as a pandemic, and it created an outrageous effect on the current healthcare and economic system throughout the globe. To date, there is no appropriate therapeutics or vaccines against the disease. The entire human race is eagerly waiting for the development of new therapeutics or vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Efforts are being taken to develop vaccines at a rapid rate for fighting against the ongoing pandemic situation. Amongst the various vaccines under consideration, some are either in the preclinical stage or in the clinical stages of development (phase-I, -II, and -III). Even, phase-III trials are being conducted for some repurposed vaccines like Bacillus Calmette-Guérin, polio vaccine, and measles-mumps-rubella. We have highlighted the ongoing clinical trial landscape of the COVID-19 as well as repurposed vaccines. An insight into the current status of the available antigenic epitopes for SARS-CoV-2 and different types of vaccine platforms of COVID-19 vaccines has been discussed. These vaccines are highlighted throughout the world by different news agencies. Moreover, ongoing clinical trials for repurposed vaccines for COVID-19 and critical factors associated with the development of COVID-19 vaccines have also been described.


Cardiodepressor and vasodepressor activities of partially purified bovine hypothalamic extract.
About 2000 bovine hypothalami were homogenized, extracted, purified and three times filtrated on Sephadex G - 25 column. During the first and second filtration the fractions with vasodepressor activity were collected for further purification. During the third filtration about 40 fractions possessing vasodepressor activity were found. The fractions emerging before vasopressin decreased the arterial blood pressure and induced transient bradycardia in rats. The results indicate that some fractions of hypothalamic extract produce vasodepressor response by vasodilatation only and the other ones by simultaneous vasodilatation and heart rate decrease.


Identification of immune-related genes from kidney and spleen of turbot, Psetta maxima (L.), by suppression subtractive hybridization following challenge with Vibrio harveyi.
Suppression subtractive hybridization was used to investigate the response of turbot, Psetta maxima (L.), to Vibrio harveyi, by using a cDNA library constructed from artificially infected turbot kidney and spleen mRNA. Forty-nine expressed sequence tags were obtained. Several immune system genes were identified, including a major histocompatibility complex (MHC) class Ia gene and a heat shock protein 70 gene. Some signalling molecules were also present in the cDNA libraries, including src-family tyrosine kinase SCK, sgk-1 serine-threonine protein kinase and amyloid precursor-like protein 2. The full length of MHC class Ia cDNA was cloned from turbot cDNA by rapid amplification of cDNA ends polymerase chain reaction. The nucleotide sequence of turbot MHC class Ia has been submitted to GenBank with accession number EF032639. The turbot MHC class Ia cDNA has an open reading frame encoding 354 amino acids, and the deduced amino acid sequence of turbot MHC class Ia has 68%, 54%, 51%, 52%, 57%, 33%, 29% and 29% identities to those of olive flounder, medaka, rainbow trout, Atlantic cod, tiger puffer, chicken, mouse and human, respectively. Quantitative reverse transcriptase-PCR was performed for the MHC class Ia gene, and it was revealed that the expression level of the MHC class Ia gene in V. harveyi-challenged turbot increased to fourfold that of the controls.


The unusual angiographic course of intracranial pseudoaneurysms.
Although rare, traumatic intracranial pseudoaneurysms remain one of the most difficult vascular lesions to diagnose and treat. A 55-year-old male patient underwent endoscopic endonasal transphenoidal resection for a pituitary macroadenoma. The operation was complicated by an arterial bleed. The initial angiogram revealed pseudoaneurysm of the anterior choroidal artery. Although the pseudoaneurysm completely disappeared on the second angiogram, it was surprisingly found to have enlarged on the third angiogram. The lesion was successfully treated with flow-diversion using a pipeline embolization device. The present case demonstrates that the natural history of iatrogenic pseudoaneurysms may be unpredictable and misleading. Traumatic pseudoaneurysms should, therefore, be carefully followed when conservative treatment is elected or when the lesion seems to have spontaneously regressed. Flow-diversion seems to be a reasonable treatment option.


IMP3 expression in serous tumors of the ovary.
Insulin-like growth factor II mRNA-binding protein (IMP3) is an oncofetal protein involved in embryogenesis, which is expressed in a variety of malignant neoplasms. It is rarely expressed in normal adult tissue and benign tumors. The aim of this study was to evaluate the expression of IMP3 in benign and malignant serous tumors of the ovary. Seventy-nine ovarian tumors were examined for IMP3 expression by immunohistochemical analysis, comprising 16 benign serous tumors, 19 borderline serous tumors, and 44 serous carcinomas. Positive staining was defined as brown staining in the cytoplasm. Negative staining was defined as absent staining or staining of <5% of tumor cells. The intensity of staining (weak, moderate, and strong) and percentage (0% to 100%) of neoplastic cells staining positive for cytoplasmic IMP3 staining were recorded in each case. Moderate to strong cytoplasmic staining for IMP3 was observed in 30 of 44 (68%) serous carcinomas of the ovary; in contrast, <5% of the borderline and benign serous tumors expressed IMP3 ranging from weak to strong cytoplasmic staining. Statistically, the difference in IMP3 expression between these groups of tumors was highly significant (P<0.0001). Our findings demonstrate moderate to strong expression of IMP3 in the majority of ovarian serous carcinomas as compared with benign/borderline serous tumors, which demonstrated weak to strong expression in a small minority (<5%) of the tumors. Thus, IMP3 may be a useful adjunctive tool in the pathologic evaluation of ovarian serous tumors.


Should preoperative or postoperative therapy be administered in the management of rectal cancer?
Combined-modality therapy consisting of surgery, radiation, and chemotherapy with 5-fluorouracil is the recommended management for patients with stage II and III rectal cancer. The place of radiation therapy as part of this combined modality treatment for localized rectal cancer will be reviewed, and some of the novel chemoradiation combinations under investigation will be highlighted. Currently, there are two sequencing approaches for the adjuvant administration of pelvic radiation therapy. The first is surgical resection and, if the tumor is stage T(3-4) and/or N(1-2), adjuvant postoperative chemoradiation and further chemotherapy. The second approach for patients with ultrasound T(3-4) or clinical T(4) disease is preoperative therapy followed by surgical resection and postoperative chemotherapy. The adjuvant radiation management approach in the United States and Europe has been shifting toward preoperative therapy to promote sphincter-preserving surgery and decrease acute and late bowel toxicity. The recently published 5-year results of the randomized German CAO/ARO/AIO 94 trial of preoperative versus postoperative chemoradiation support a standard preoperative treatment approach. Preoperative chemoradiation therapy also allows for the investigation of innovative agents (capecitabine, oxaliplatin, irinotecan, bevacizumab, and cetuximab) in combination with pelvic radiation. These new combinations may have the potential to further increase the therapeutic benefit of neoadjuvant therapy.


Messenger RNA-based therapeutics for brain diseases: An animal study for augmenting clearance of beta-amyloid by intracerebral administration of neprilysin mRNA loaded in polyplex nanomicelles.
Alzheimer's disease (AD) pathogenesis is considered to be the metabolic imbalance between anabolism and clearance of amyloid-beta (Aβ), and the strategy of breaking the equilibrium between soluble and insoluble forms of Aβ is likely to help prevent the progression of AD. Neprilysin (NEP) plays a major role in the clearance of Aβ in the brain, and its supplementation using viral vectors has shown to decrease Aβ deposition and prevent pathogenic changes in the brain. In this study, we developed a new therapeutic strategy by mRNA-based gene introduction. mRNA has the advantages of negligible risk of random integration into genome and not needing to be transcribed precludes the need for nuclear entry. This allows efficient protein expression in slowly-dividing or non-dividing cells, such as neural cells. We constructed mRNA encoding the mouse NEP protein and evaluated its ability degrade Aβ. In vitro transfection of NEP mRNA to primary neurons exhibited Amyloid Precursor Protein (APP) degradation activity superior to that of NEP encoding plasmid DNA. We then evaluated the in vivo activity of NEP mRNA by intracerebroventricular (i.c.v.) infusion using a cationic polymer-based PEGylated nanocarrier to form polyplex nanomicelles, which have been shown to have a high potential to deliver mRNA to various target tissues and organs. Nanomicelles carrying a GFP-NEP fusion mRNA produced efficient protein expression in a diffuse manner surrounding the ventricular space. An ELISA evaluation revealed that the mRNA infusion significantly augmented NEP level and effectively reduced the concentration of Aβ that had been supplemented in the mouse brain. To the best of our knowledge, this is the first study to demonstrate the therapeutic potential of introducing exogenous mRNA for the treatment of brain diseases, opening the new era of mRNA-based therapeutics.


Subsequent products after antioxidant actions of beta-carotene and alpha-tocopherol have no Salmonella mutagenicity.
Beta-carotene and alpha-tocopherol are important antioxidants biologically, but whether their oxidized products are toxic or not remains to be discovered. Here, we chromatographically separated 5 pure products or isomeric mixtures from reaction mixtures of beta-carotene and reactive oxygens, and 17 lipid-radical scavenging products of alpha-tocopherol. The products were tested for mutagenicity using Salmonella typhimurium TA98, TA100, TA102, and TA104, in the presence and absence of S9. None showed mutagenicity against any of the four strains, or cytotoxicity that influenced the survival of the bacteria. Lipid-peroxides have been known to increase the formation of mutagens from dietary procarcinogens such as heterocyclic amines. So, we also measured the activity to increase 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) mutagenicity. The products from beta-carotene and alpha-tocopherol did not increase, but rather several of them suppressed, the mutagenicity of Trp-P-2. Thus, the products of beta-carotene and alpha-tocopherol formed after the antioxidant actions were not genotoxic.


Compulsive "helpfulness": or, how I learned to stop working so hard and love the group.
Compulsive helpfulness, an anxiety-laden need by the therapist to feel helpful, akin to the notion of rescuing others, is a potential pitfall in developing an effective psychotherapy group. It can be regarded variously: (1) as a reaction formation against feelings of boredom and frustration stimulated by such phenomena as group resistance or the enactment of inauthentic relationships in the group; (2) as a therapist style driven by a transferential reaction to be regarded as competent and worthy; (3) as an induced countertransference enactment tied to group members' frustration and passivity about their own interpersonal inadequacies; (4) and as a manic defense against despair over the feeling that one's loving has not done any good. In this essay, the author explores his struggle to identify and come to terms with compulsive helpfulness as a dominant theme in the early stages of his tenure as leader of a psychotherapy group.


Racial/Ethnic and nativity differences in birth outcomes among mothers in New York City: the role of social ties and social support.
Immigrants have lower rates of low birth weight (LBW) and to some extent preterm birth (PTB), than their US-born counterparts. This pattern has been termed the 'immigrant health paradox'. Social ties and support are one proposed explanation for this phenomenon. We examined the contribution of social ties and social support to LBW and PTB by race/ethnicity and nativity among women in New York City (NYC). The NYC Pregnancy Risk Assessment Monitoring System survey (2004-2007) data, linked with the selected items from birth certificates, were used to examine LBW and PTB by race/ethnicity and nativity status and the role of social ties and social support to adverse birth outcomes using bivariate and multivariable analyses. SUDAAN software was used to adjust for complex survey design and sampling weights. US- and foreign-born Blacks had significantly increased odds of PTB [adjusted odds ratio (AOR) = 2.43, 95 % CI 1.56, 3.77 and AOR = 2.6, 95 % CI 1.66, 4.24, respectively] compared to US-born Whites. Odds of PTB among foreign-born Other Latinas, Island-born Puerto Ricans' and foreign-born Asians' were not significantly different from US-born Whites, while odds of PTB for foreign-born Whites were significantly lower (AOR = 0.47, 95 % CI 0.26, 0.84). US and foreign-born Blacks' odds of LBW were 2.5 fold that of US-born Whites. Fewer social ties were associated with 32-39 % lower odds of PTB. Lower social support was associated with decreased odds of LBW (AOR 0.69, 95 % CI 0.50, 0.96). We found stronger evidence of the immigrant health paradox across racial/ethnic groups for PTB than for LBW. Results also point to the importance of accurately assessing social ties and social support during pregnancy and to considering the potential downside of social ties.


HTLV-III large envelope protein (gp120) suppresses PHA-induced lymphocyte blastogenesis.
The addition of inactivated preparations of purified human T cell lymphotropic virus (HTLV-III) was found to inhibit normal human lymphocyte phytohemagglutinin (PHA)-induced blastogenesis but had no effect on concanavalin A (Con A), pokeweek mitogen, or allogeneic stimulation. The inhibition was concentration-dependent and also dependent on adding the virus preparation before or at the same time as PHA. The CD4 molecule is the receptor for HTLV-III binding. Immunopurified large envelope protein (gp120) from HTLV-III was found to bind to the CD4 molecule and also inhibited PHA-induced lymphocyte blastogenesis. These results suggest that the gp120 viral protein may alter immune function by binding to the CD4 molecule, which in turn serves as an "off" signal to lymphocyte response to PHA stimulation. Alternatively, by binding to the CD4 molecule, gp120 may interfere with the interaction of this molecule with class II histocompatibility antigens on accessory cells, thus selectively suppressing PHA response.


The DYW Subgroup PPR Protein MEF35 Targets RNA Editing Sites in the Mitochondrial rpl16, nad4 and cob mRNAs in Arabidopsis thaliana.
RNA editing in plant mitochondria and plastids alters specific nucleotides from cytidine (C) to uridine (U) mostly in mRNAs. A number of PLS-class PPR proteins have been characterized as RNA recognition factors for specific RNA editing sites, all containing a C-terminal extension, the E domain, and some an additional DYW domain, named after the characteristic C-terminal amino acid triplet of this domain. Presently the recognition factors for more than 300 mitochondrial editing sites are still unidentified. In order to characterize these missing factors, the recently proposed computational prediction tool could be of use to assign target RNA editing sites to PPR proteins of yet unknown function. Using this target prediction approach we identified the nuclear gene MEF35 (Mitochondrial Editing Factor 35) to be required for RNA editing at three sites in mitochondria of Arabidopsis thaliana. The MEF35 protein contains eleven PPR repeats and E and DYW extensions at the C-terminus. Two T-DNA insertion mutants, one inserted just upstream and the other inside the reading frame encoding the DYW domain, show loss of editing at a site in each of the mRNAs for protein 16 in the large ribosomal subunit (site rpl16-209), for cytochrome b (cob-286) and for subunit 4 of complex I (nad4-1373), respectively. Editing is restored upon introduction of the wild type MEF35 gene in the reading frame mutant. The MEF35 protein interacts in Y2H assays with the mitochondrial MORF1 and MORF8 proteins, mutation of the latter also influences editing at two of the three MEF35 target sites. Homozygous mutant plants develop indistinguishably from wild type plants, although the RPL16 and COB/CYTB proteins are essential and the amino acids encoded after the editing events are conserved in most plant species. These results demonstrate the feasibility of the computational target prediction to screen for target RNA editing sites of E domain containing PLS-class PPR proteins.


Surgical treatment for Parkinson's disease.
Since the early 1930s, physicians have developed and refined various surgical therapies for the treatment of Parkinson's disease. In this review we examine some of the problems associated with early surgical therapies, the development of new techniques and targets, and the results of clinical trials examining the safety and efficacy of these techniques. Ablative techniques include pallidotomy, thalamotomy, and, more recently, subthalamotomy. Because of concern over the high incidence of side-effects associated with bilateral ablative procedures, alternative approaches were explored. Deep brain stimulation (DBS) was subsequently developed and successfully applied in the internal globus pallidus, subthalamic nucleus, and thalamus for the treatment of Parkinson's disease. Recent approaches include biological neurorestorative techniques--surgical therapies with transplantation, gene therapy, and growth factors are all being studied. Although a great deal of work remains to be done, advances in surgical therapies for the treatment of Parkinson's disease are moving forward at an unprecedented pace.


Hormone responsiveness of isolated catfish hepatocytes in perifusion system is higher than in flasks incubation.
Hepatocytes were isolated from catfish (lctalurus melas) by conventional collagenase digestion. Sensitivities of liver cells isolated from the same fish to the glycogenolytic action of epinephrine, mammalian glucagon, catfish glucagon, catfish glucagon-like peptide, synthetic fragment 19-29 of anglerfish glucagon I, fragment 19-29 of anglerfish glucagon II, and anglerfish glucagon II were compared in two different systems: perifusion in a Bio-Gel P4 column and flask incubation. Both experimental procedures were continued for a total of 100-120 min, while hormones were applied simultaneously to both preparations for 10 min. Effluent fractions from the columns and incubation media from the flasks were collected for glucose determination. The hormonal effects were clearly enhanced in perifused cells compared to those in cells incubated in flasks, the effect being especially evident at physiological concentrations of hormones. The hormonal effects in both systems were dose-dependent. Epinephrine and mammalian glucagon (10 nM), applied separately to the same column, produced two different peaks, glucagon causing more glucose production than epinephrine. In the presence of 0.4 mM glucose in the perifusion system, hormonal effects were diminished, implying that glucose accumulation during incubation of liver cells in flasks might affect hormonal effects. The results obtained in this study indicate that piscine hepatocytes suspended and perifused in a Bio-Gel column are more sensitive to physiological concentrations of glycogenolytic hormones and may represent a new tool for experimental studies of fish liver metabolism and its hormonal regulation.


Role of hydroxyurea during structured treatment interruptions.
Highly active antiretroviral therapies (HAART) represent a major advance in the treatment of HIV infection. Although with HAART a substantial suppression of viral replication can be obtained, eradication of the virus from the body cannot be achieved. Therefore, HIV-infected subjects have to be treated for the rest of their lives. Long term treatment will increase the frequency of: i) drug-related side effects; ii) onset of drug-resistant viral strains; iii) non-adherence of the patients to the treatment. Structured treatment interruptions (STI)-HAART might represent a feasible alternative and preliminary studies have shown that STI-HAART might induce immune control in patients treated in the early stage of infection. This regimen does not produce similar effects in patients treated during the chronic phase of the infection. However, there are some clinical data suggesting a possible role of hydroxyurea (HU) in inducing control of HIV replication in patients with established infection. In this manuscript in vitro and in vivo data indicating that HU might play a major role in the setting of STI-HAART will be presented.


Estradiol-Induced Epigenetically Mediated Mechanisms and Regulation of Gene Expression.
Gonadal hormone 17β-estradiol (E2) and its receptors are key regulators of gene transcription by binding to estrogen responsive elements in the genome. Besides the classical genomic action, E2 regulates gene transcription via the modification of epigenetic marks on DNA and histone proteins. Depending on the reaction partner, liganded estrogen receptor (ER) promotes DNA methylation at the promoter or enhancer regions. In addition, ERs are important regulators of passive and active DNA demethylation. Furthermore, ERs cooperating with different histone modifying enzymes and chromatin remodeling complexes alter gene transcription. In this review, we survey the basic mechanisms and interactions between estrogen receptors and DNA methylation, demethylation and histone modification processes as well as chromatin remodeling complexes. The particular relevance of these mechanisms to physiological processes in memory formation, embryonic development, spermatogenesis and aging as well as in pathophysiological changes in carcinogenesis is also discussed.


Residual subjective daytime sleepiness under CPAP treatment in initially somnolent apnea patients: a pilot study using data mining methods.
Despite correct treatment with positive airway pressure (PAP), obstructive sleep apnea (OSA) patients sometimes remain subjectively somnolent. The reliability of the Epworth Sleepiness Scale (ESS) has been established for healthy subjects and patients under stable conditions; the ESS may eventually vary among treated OSA patients, biasing the results of a cross-sectional analysis of persisting sleepiness. The objective of this study was to depict the evolution of subjective vigilance under treatment using an index of ESS variability (DeltaESS). In 80 OSA patients (apnea-hypopnea index [AHI]=54+/-26/h), initially somnolent (ESS=15+/-3) and treated with auto-titrating PAP (APAP) (oxyhaemoglobin desaturation index 3% [ODIapap]=3.4+/-2.2/h; daily APAP use=5.3+/-1.5 h) during 434+/-73 days, ESS scores were regularly collected four times every 109+/-36 days. DESS was calculated and data mining methods (Segmentation and Decision Tree) were used to determine homogeneous groups according to the evolution of ESS scores. When assessed cross-sectionally, 14-25% of the subjects were recognized as somnolent, depending on the moment when ESS was administered. Using data mining methods, three groups were clearly identifiable: two without residual somnolence - group 1, n=38 (47%), with high DeltaESS=-2.9+/-0.8, baseline ESS=16.3+/-3.3, AHI=58.5+/-26.1/h, mean ESSapap=5.1+/-2.4 and group 2, n=31 (39%), with low DeltaESS=-1.1+/-0.5, baseline ESS=13.2+/-1.4, AHI=53+/-27.3/h, mean ESSapap=8.8+/-1.9; and one with persisting sleepiness; group 3, n=11 (14%), with low DeltaESS=-0.3+/-0.8, baseline ESS=16.3+/-3, AHI=38.7+/-10.8/h, mean ESSapap=14.1+/-1.9. Compliance to PAP was high and comparable in the three groups. Age and body mass index (BMI) did not differ. Data mining methods helped to identify 14% of subjects with persisting sleepiness. Validation needs to be done on a larger population in order to determine predictive rules.


A case of a long-term survival achieved by surgical treatment and chemotherapy for late recurrence of AFP-producing gastric cancer.
Alpha-fetoprotein-producing gastric cancer (AFP-GC) is a relatively rare disease, with a dismal prognosis. We report the case of a patient with long-term survival after surgery for the recurrence of AFP-GC. A 71-year-old man was diagnosed with gastric cancer and underwent distal gastrectomy with D2 lymphadenectomy (pT3N2M0). Pathological examination of the resected specimen revealed AFP-GC. Fifteen years after the gastrectomy, the patient experienced anorexia and was admitted with a mass located at the mesentery of the small intestine. Following a diagnosis of gastrointestinal stromal tumor of the mesentery, a tumor resection with partial small intestine was performed. The final histopathological diagnosis was AFP-GC's recurrence in the small-bowel mesentery. Two months later, multiple liver metastases were identified, and serum AFP level was found to be extremely high (17,447 ng/mL). Chemotherapy with S-1+CDDP (SP) was initiated for liver metastasis. However, owing to anorexia and fatigue, SP therapy was discontinued following the patient's request at the end of two courses. A CT scan at 1 month after the discontinuation of chemotherapy did not reveal liver metastasis, and serum AFP level decreased to the normal range. He is alive at present with no re-recurrence and no elevation of serum AFP level at 7 years after the second surgery without any chemotherapy. Even if recurrence of AFP-GC is diagnosed, radical resection and chemotherapy are effective, as noted in the present case.


Deterrence of birds with an artificial predator, the RobotFalcon.
Collisions between birds and airplanes can damage aircrafts, resulting in delays and cancellation of flights, costing the international civil aviation industry more than 1.4 billion US dollars annually. Driving away birds is therefore crucial, but the effectiveness of current deterrence methods is limited. Live avian predators can be an effective deterrent, because potential prey will not habituate to them, but live predators cannot be controlled entirely. Thus, there is an urgent need for new deterrence methods. We developed the RobotFalcon, a device modelled after the peregrine falcon, and tested its effectiveness to deter flocks of corvids, gulls, starlings and lapwings. We compared its effectiveness with that of a drone, and of conventional methods routinely applied at a military airbase. The RobotFalcon scared away bird flocks from fields immediately, and these fields subsequently remained free of bird flocks for hours. The RobotFalcon outperformed the drone and the best conventional method at the airbase (distress calls). Importantly, there was no evidence that bird flocks habituated to the RobotFalcon over the course of the fieldwork. We conclude that the RobotFalcon is a practical and ethical solution to drive away bird flocks with all advantages of live predators but without their limitations.


Damage modeling and statistical analysis of optics damage performance in MJ-class laser systems.
Modeling the lifetime of a fused silica optic is described for a multiple beam, MJ-class laser system. This entails combining optic processing data along with laser shot data to account for complete history of optic processing and shot exposure. Integrating with online inspection data allows for the construction of a performance metric to describe how an optic performs with respect to the model. This methodology helps to validate the damage model as well as allows strategic planning and identifying potential hidden parameters that are affecting the optic's performance.


[Granulomatous myopathy in patients with sarcoidosis and myasthenia gravis].
Granulomatous myopathies are extremely rare. The finding of epithelioid granulomas in muscle biopsy indicates mostly an involvement of the skeletal muscle in systemic sarcoidosis. In this report we provide description of five patients with previously diagnosed sarcoidosis (pulmonary or cutaneous), in which a clinically significant muscle weakness developed. We aim at demonstrating the value of muscle biopsy for further treatment of the patients, since clinically indistinguishable myopathies can arise not due to the involvement of muscles in the granulomatous process, but due to the corticosteroid-induced changes (chronic steroid myopathy). The demonstration of a selective atrophy of type II muscle fibers can provide the clue for distinguishing the patients, in which the corticosteroid treatment should continue, from those, in which the treatment should be modified. Further, we discuss a rare finding of granulomas in muscle biopsies of two patients with myasthenia gravis (MG) associated with thymoma. Although it is difficult to explain the pathogenesis of this event, MG should be considered in the differential diagnosis of granulomatous myopathies. Moreover, the finding of granulomas along with lymphocytic infiltration in MG muscle should lead to a search for an underlying thymic neoplasm.


Infrared and Raman spectra of lignin substructures: Coniferyl alcohol, abietin, and coniferyl aldehyde.
Anatomical and chemical information can be linked by Raman imaging. Behind every pixel of the image is a Raman spectrum, which contains all the information as a molecular fingerprint. Yet to understand the spectra, the bands have to be assigned to components and their molecular structures. Although the lignin distribution is easily tracked in plant tissues, the assignment of the spectra is not good enough to allow in-depth analysis of the composition. Assignments of three lignin model compounds were derived from polarization measurements and quantum-chemical computations. Raman spectra of coniferyl alcohol crystals showed orientation dependence, which helped in band assignment. Abietin showed a Raman spectrum that was very similar to the spectrum of coniferyl alcohol, whereas its IR spectrum was very different due to bands of the sugar moiety. The Raman spectrum of coniferyl aldehyde is affected by the crystal order of molecules. All three compounds show much stronger band intensities than unconjugated single aromatic rings, indicating that the bulk of the lignin structure has significantly reduced contribution to Raman band intensities. Therefore, it is possible to highlight certain structures of lignin with Raman spectroscopy, because low amounts of a compound do not necessarily mean weak features in the spectrum.


Role of cytoskeleton in canalicular contraction in cultured differentiated hepatocytes.
We and others have published data that indicate that the role played by microtubules and microfilaments in biliary secretion is as follows: microtubules play a part in secretion and microfilaments play a part in the canalicular contraction. To further study the role of the cytoskeleton in canalicular contraction, we observed the contraction of bile canaliculi (BC) induced by vasopressin (VP) in cultured differentiated hepatocytes treated with several agents that selectively rearrange the cytoskeleton. The hepatocytes obtained from 14-day-old rats were cultured for 48 hours. The BC formed between the cells were dilated and closely sealed by junctional complexes. Ruthenium red stain showed that the junctional complexes of the BC were tightly sealed. Cytoskeletal changes were observed by double-labeled fluorescence microscopy. A spontaneous contraction of the BC was rarely seen during a 60-minute observation period in controls. When the hepatocytes were incubated with VP (10(-8) M), the canalicular contraction began at 30 minutes, gradually progressed, and was complete by 60 minutes. The contraction was reversed after 60 minutes of incubation in VP-free medium. In cytochalasin B-treated hepatocytes, actin appeared to form pools around the dilated BC, and the canalicular contraction after VP was rarely observed. In colchicine-treated hepatocytes, the microtubules were depolymerized. Although the BC appeared unaffected by colchicine alone, the canalicular contraction induced by VP was markedly decreased. beta-lumicolchicine had no effect on the cytoskeleton or on the canalicular contraction. Mg2(+)-ATPase histochemistry revealed that the BC that did not contract after VP contained little Mg2(+)-ATPase reaction product. When the BC contracted, diverticula came off to form diacytotic vesicles, as indicated by the presence of the BC marker enzyme reaction product within the vesicles. Colchicine treatment blocked the diacytotic process. This prevented the contraction stimulated by VP, because all of the routes of escape of the canalicular contents were blocked off, including diacytosis. In conclusion, the integrity of actin filaments and Mg2(+)-ATPase is necessary for VP-induced contraction, and the integrity of microtubules is essential for regurgitation of BC content (diacytosis).


Alterations in peripheral blood leucocyte distribution in response to local inflammatory stimuli in the rat.
Changes in peripheral blood leucocyte distribution were monitored following subcutaneous injections of turpentine and subcutaneous and intrapleural injections of dextran. The responses in all three groups were similar; marked increases in total counts occurred at 6 hr, due largely to significant increases in neutrophil counts. Increases in lymphocyte counts also occurred and were more marked in the dextran-treated animals. By 12 hr, total counts fell, due to drops in both neutrophil and lymphocyte counts. In both dextran groups, neutrophil counts rose again between 24 and 72 hr, but in the turpentine group the fall persisted until 96 hr before counts reached a second peak at 14 days. In all groups lymphocytes showed an increase again at 18-72 hr. The rise in neutrophil counts observed 6 hr after the injection of turpentine was shown to be due principally to premature release of neutrophils from the bone marrow maturation compartment. Depletion of this compartment resulted in a drop in numbers of circulating neutrophils between 6 and 12 hr, in spite of the continued release of progressively less mature cells. The recovery in neutrophil counts between 72 and 96 hr was thought to be the expression of an increase in the rate of stem cell division. The continued rise to a second peak at 7 days probably resulted from increased marrow cellularity. It is suggested that the fluctuations in peripheral blood leucocyte counts may influence the initial colonisation of the local inflammatory lesion. The increase in neutrophil counts at 6 hr corresponds with the time of maximum neutrophil emigration reported by other workers and the fall in counts at 12 hr corresponds with the time of decreased neutrophil emigration.


Wavelength-locking-free 1.57µm differential absorption lidar for CO₂ sensing.
We propose a novel wavelength-locking-free differential absorption lidar system for CO₂ sensing. The ON-line wavelength laser was wavelength modulated around a specific CO₂ absorption line to ensure that the emission from the ON-line laser hit the atmospheric CO₂ absorption line peak twice a cycle. In the meantime, the intensity of the ON-line and OFF-line wavelength lasers were sinusoidally intensity modulated to enhance the SNR of the back-scattered signal. As a consequence, the system configuration was simplified and the measurement error caused by the deviation of CO₂ absorption coefficient from the long-time ON-line wavelength drifting was completely eliminated. Furthermore, a more precise calibration method was developed which could simultaneously calibrate the offset and precision of the lidar detector. This method could be applied to other differential-absorption-based lidar systems. The result showed that a measurement precision of 0.525% for the column concentration was achieved in 1 s time interval through a path of 780m. We recorded the CO₂ concentration variation for 12 hours starting from mid-night, the result showed that the course of the concentration derived from the DIAL was in good agreement with that of the in situ CO₂ sensor only when the status of atmosphere was stable.


[Agents for sedation and analgesia in the intensive care unit].
Sedation-analgesia for critically ill patients is usually performed with the combination of a sedative agent and an opioid. Midazolam and propofol are the agents most commonly used for sedation in ICU. The quality of the sedation is quite comparable with both agents, but pharmacokinetic properties of propofol allow a more rapid weaning process from mechanical ventilation. However, implementation of algorithms to adjust drug dosages reduces ventilator days and limits the kinetic differences between propofol and midazolam. Among the adverse events associated with propofol, propofol infusion syndrome is a rare but lethal aspect of propofol therapy. Opioids are the mainstay of analgesic therapy. They interact synergistically with hypnotics. Sufentanil, fentanyl and morphine are the most frequently used opioids. Remifentanil is an ultrashort acting opiate that does not appear to accumulate with prolonged use. The advent of remifentanil has allowed the use of analgesia-based sedation.


[Therapeutic arsenal in adult epilepsy].
Over the last 20 years, the number of drugs licensed for the treatment of seizure disorders has increased exponentially. Although the second-generation antiepileptic drugs have failed to significantly reduce the proportion of drug-resistant patients, their safety profile and the reduction of drug to drug interactions now allow to better tailor the treatment to each patient. The decision to initiate an antiepileptic drug in newly diagnosed epilepsy relies on a careful assessment of the expected benefit according both to the risk of seizure recurrence and to the risk of drug-related adverse events. In patients in whom a treatment is initiated, the final choice of the drug must then take into account both the expected efficacy of the drug and its relevance to the specific characteristics of the patient, including his/her gender, age or comorbidities. This approach is essential in all patients, with a special attention to the issues raised in women of childbearing age.


Bioinformatics led discovery of biomarkers related to immune infiltration in diabetes nephropathy.
The leading cause of end-stage renal disease is diabetic nephropathy (DN). A key factor in DN is immune cell infiltration (ICI). It has been shown that immune-related genes play a significant role in inflammation and immune cell recruitment. However, neither the underlying mechanisms nor immune-related biomarkers have been identified in DNs. Using bioinformatics, this study investigated biomarkers associated with immunity in DN. Using bioinformatic methods, this study aimed to identify biomarkers and immune infiltration associated with DN. Gene expression profiles (GSE30528, GSE47183, and GSE104948) were selected from the Gene Expression Omnibus database. First, we identified 23 differentially expressed immune-related genes and 7 signature genes, LYZ, CCL5, ALB, IGF1, CXCL2, NR4A2, and RBP4. Subsequently, protein-protein interaction networks were created, and functional enrichment analysis and genome enrichment analysis were performed using the gene ontology and Kyoto Encyclopedia of Genes and Genome databases. In the R software, the ConsensusClusterPlus package identified 2 different immune modes (cluster A and cluster B) following the consistent clustering method. The infiltration of immune cells between the 2 clusters was analyzed by applying the CIBERSORT method. And preliminarily verified the characteristic genes through in vitro experiments. In this study, the samples of diabetes nephropathy were classified based on immune related genes, and the Hub genes LYZ, CCL5, ALB, IGF1, CXCL2, NR4A2 and RBP4 related to immune infiltration of diabetes nephropathy were obtained through the analysis of gene expression differences between different subtypes. This study was based on bioinformatics technology to analyze the biomarkers of immune related genes in diabetes nephropathy. To analyze the pathogenesis of diabetes nephropathy at the RNA level, and ultimately provide guidance for disease diagnosis, treatment, and prognosis.


PHYSICAL THERAPY MANAGEMENT OF ICE HOCKEY ATHLETES: FROM THE RINK TO THE CLINIC AND BACK.
The increasing number of athletes playing hockey compels rehabilitation professionals working in orthopedic and sports settings to understand the unique functional demands of ice hockey and the patterns of injuries they may promote. The purpose of this clinical perspective is to: (1) discuss the functional implications of different positions and age levels on injury prevalence within the sport; (2) summarize the seven most common injuries sustained by ice hockey athletes; and (3) present a conceptual model for the clinical management and prevention of these injuries by rehabilitation professionals. A narrative review and synthesis was conducted of currently available literature on prevalence, etiology, rehabilitative intervention, prognosis, and prevention of ice hockey injuries. Research evidence is available to support the prevalence of injuries sustained while participating in ice hockey, as well as the most effective clinical treatment protocols to treat them. Most of the existing protocols are based on clinical and sports experience with incorporation of scientific data. This clinical commentary reviews the current concepts of ice hockey injury care and prevention, based on scientific information regarding the incidence, mechanism, rehabilitation protocols, prognosis, and prevention of injuries. Science-based, patient-centered reasoning is integral to provide the highest quality of rehabilitative and preventative care for ice hockey athletes by physical therapists. 5.


[Plasma amino-terminal pro-brain natriuretic peptide level and affecting factors in a community-based healthy Chinese population].
To investigate the plasma level of amino-terminal pro-brain natriuretic peptide (NT-proBNP) and related influencing factors in a community-based healthy population in Beijing area. We measured plasma NT-proBNP level by fluoroimmunoassay between March 2012 and July 2012 from 1 034 healthy subjects (including 486 men and 548 women). Empiric method was used to determine the reference value and influencing factors were analyzed. Age and gender are important factors affecting the level of NT-proBNP in healthy subjects. NT-proBNP plasma level is significantly higher in women than in men within each age strata below 75 years old, i.e. < 45, 45-54, 55-64 and 65-74 years old (P = 0.005, 0.001, 0.001, 0.011 respectively), but NT-proBNP plasma level is similar between male and female older than 75 years (P = 0.504). NT-proBNP level also increases with age irrespective of gender. Body mass index (BMI) is another independent influencing factor of NT-proBNP (P < 0.001), while estimated glomerular filtration rate is not influencing factor. The reference range of NT-proBNP is < 133 ng/L for men and < 289 ng/L for women aged < 55 years old, < 185 ng/L for men and < 333 ng/L for women aged between 55 and 64 years old, and < 465 ng/L for men and < 378 ng/L for women aged ≥ 75 years old. The major influencing factors of NT-proBNP level in the healthy population are age, gender and BMI. It essential to establish normal reference range of NT-proBNP according to these factors for Chinese population.


Elevation of Transient Receptor Potential Vanilloid 1 Function in the Lateral Habenula Mediates Aversive Behaviors in Alcohol-withdrawn Rats.
Chronic alcohol use and withdrawal leads to increased pain perception, anxiety, and depression. These aberrant behaviors are accompanied by increased excitatory glutamatergic transmission to, and activity of, the lateral habenula neurons.Vanilloid type 1, or TRPV1, channels are expressed in the habenula and they facilitate glutamatergic transmission. Whether TRPV1 channel plays a role in habenula hyperactivity is not clear. Glutamatergic transmission in the lateral habenula was inhibited by TRPV1 channel antagonists. In vivo, local administration of TRPV1 antagonists into the lateral habenula attenuated hyperalgesia, anxiety, and relapse-like drinking in rats who chronically consumed alcohol.The data suggest that enhanced TRPV1 channel function during withdrawal may contribute to aberrant behavior that promotes relapse alcohol consumption. Recent rat studies indicate that alcohol withdrawal can trigger a negative emotional state including anxiety- and depression-like behaviors and hyperalgesia, as well as elevated glutamatergic transmission and activity in lateral habenula neurons. TRPV1, a vanilloid receptor expressed in the habenula, is involved in pain, alcohol dependence, and glutamatergic transmission. The authors therefore hypothesized that TRPV1 contributes to the changes in both the behavioral phenotypes and the habenula activity in alcohol-withdrawn rats. Adult male Long-Evans rats (n = 110 and 280 for electrophysiology and behaviors, respectively), randomly assigned into the alcohol and water (Naïve) groups, were trained to consume either alcohol or water-only using an intermittent-access procedure. Slice electrophysiology was used to measure spontaneous excitatory postsynaptic currents and firing of lateral habenula neurons. The primary outcome was the change in alcohol-related behaviors and lateral habenula activity induced by pharmacologic manipulation of TRPV1 activity. The basal frequency of spontaneous excitatory postsynaptic currents and firing of lateral habenula neurons in alcohol-withdrawn rats was significantly increased. The TRPV1 antagonist capsazepine (10 µM) induced a stronger inhibition on spontaneous excitatory postsynaptic currents (mean ± SD; by 26.1 ± 27.9% [n = 11] vs. 6.7 ± 18.6% [n = 17], P = 0.027) and firing (by 23.4 ± 17.6% [n = 9] vs. 11.9 ± 16.3% [n = 12], P = 0.025) in Withdrawn rats than Naive rats. By contrast, the TRPV1 agonist capsaicin (3 μM) produced a weaker potentiation in Withdrawn than Naïve rats (spontaneous excitatory postsynaptic currents: by 203.6 ± 124.7% [n = 20] vs. 415.2 ± 424.3% [n = 15], P < 0.001; firing: 38.1 ± 14.7% [n = 11] vs. 73.9 ± 41.9% [n = 11], P < 0.001). Conversely, capsaicin's actions in Naïve but not in Withdrawn rats were significantly attenuated by the pretreatment of TRPV1 endogenous agonist N-Oleoyldopamine. In Withdrawn rats, intra-habenula infusion of TRPV1 antagonists attenuated hyperalgesia and anxiety-like behaviors, decreased alcohol consumption upon resuming drinking, and elicited a conditioned place preference. Enhanced TRPV1 function may contribute to increased glutamatergic transmission and activity of lateral habenula neurons, resulting in the aberrant behaviors during ethanol withdrawal.


Osteoblast differentiation and migration are regulated by dynamin GTPase activity.
Bone formation is controlled by osteoblasts, but the signaling proteins that control osteoblast differentiation and function are still unclear. We examined if the dynamin GTPase, which is associated with actin remodeling and migration in other cells, plays a role in osteoblast differentiation and migration. Dynamin mRNA was expressed in primary osteoblasts throughout differentiation (0-21 days). However, alkaline phosphatase (ALP) activity, a marker of osteoblast differentiation, was decreased in osteoblasts over-expressing dynamin. Conversely, ALP activity was increased following shRNA-mediated knockdown of dynamin and in osteoblasts treated with the dynamin inhibitor, dynasore. Dynasore also reduced c-fos and osterix expression, markers of early osteoblasts, suggesting a role for dynamin in pre-osteoblast to osteoblast differentiation. Since dynamin GTPase activity is regulated by tyrosine phosphorylation, we examined the mechanism of dynamin dephosphorylation in osteoblasts. Dynamin formed a protein complex with the tyrosine phosphatase PTP-PEST and inhibition of phosphatase activity increased the level of phosphorylated dynamin. Further, PTP-PEST blocked the Src-mediated increase in the phosphorylation and GTPase activity of wild-type dynamin but not the phosphorylation mutant dynY231F/Y597F. Although ALP activity was increased in osteoblasts expressing GTPase-defective dynK44A, and to a lesser extent dynY231F/Y597F, osteoblast migration was significantly inhibited by dynK44A and dynY231F/Y597F. These studies demonstrate a novel role for dynamin GTPase activity and phosphorylation in osteoblast differentiation and migration, which may be important for bone formation.


Synergistic induction of apoptosis by combination treatment with mesupron and auranofin in human breast cancer cells.
Urokinase-type plasminogen activator (uPA) has been validated as a predictive or prognostic biomarker protein, and mesupron is considered the first-in-class anticancer agent to inhibit uPA activity in human breast cancer. In the present study, we showed that the synergism between mesupron and auranofin, a thioredoxin reductase inhibitor, for inducing of apoptosis in MCF-7 human breast cancer cells. Our results demonstrated that mesupron and auranofin significantly lead to inhibition of the cancer cells proliferation; cell cycle arrest at the G1/S phase of the cell cycle, and apoptosis as indicated by caspase 3 activation, poly(ADP-ribose) polymerase cleavage, and annexin V staining. Isobologram analyses of MCF-7 cells showed a clear synergism between mesupron and auranofin. This combined treatment decreased the levels of mitochondrial anti-apoptotic factors, such as BCL-2, BCL-xL, and MCL-1 and caused nuclear translocation of apoptosis-inducing factor. Mitochondrial membrane potential (Δψ m ) was found to be strongly disrupted in combination-treated cells. In addition, combination treatment significantly enhanced the overproduction of reactive oxygen species, which was rescued by N-acetylcysteine treatment. The combination treatment suppressed phosphorylation of Akt, thus contributing to apoptosis. Taken together, our data suggest that the use of mesupron in combination with auranofin may be important in achieving high anticancer synergy.


Biochemical and structural characterization of the paralogous benzoate CoA ligases from Burkholderia xenovorans LB400: defining the entry point into the novel benzoate oxidation (box) pathway.
Xenobiotic aromatic compounds represent one of the most significant classes of environmental pollutants. A novel benzoate oxidation (box) pathway has been identified recently in Burkholderia xenovorans LB400 (referred to simply as LB400) that is capable of assimilating benzoate and intimately tied to the degradation of polychlorinated biphenyls (PCBs). The box pathway in LB400 is present in two paralogous copies (boxM and boxC) and encodes eight enzymes with the first committed step catalyzed by benzoate CoA ligase (BCL). As a first step towards delineating the biochemical role of the box pathway in LB400, we have carried out functional studies of the paralogous BCL enzymes (BCLM and BCLC) with 20 different putative substrates. We have established a structural rationale for the observed substrate specificities on the basis of a 1.84 A crystal structure of BCLM in complex with benzoate. These data show that, while BCLM and BCLC display similar overall substrate specificities, BCLM is significantly more active towards benzoate and 2-aminobenzoate with tighter binding (Km) and a faster reaction rate (Vmax). Despite these clear functional differences, the residues that define the substrate-binding site in BCLM are completely conserved in BCLC, suggesting that second shell residues may play a significant role in substrate recognition and catalysis. Furthermore, comparison of the active site of BCLM with the recently solved structures of 4-chlorobenzoate CoA ligase and 2, 3-dihydroxybenzoate CoA ligase offers additional insight into the molecular features that mediate substrate binding in adenylate-forming enzymes. This study provides the first biochemical characterization of a Box enzyme from LB400 and the first structural characterization of a Box enzyme from any organism, and further substantiates the concept of distinct roles for the two paralogous box pathways in LB400.


Fungal and bacterial community dynamics in substrates during the cultivation of morels (Morchella rufobrunnea) indoors.
Morel mushrooms (Morchella, Pezizales) are highly prized edible fungi. Approaches to cultivate morels indoors in pasteurized composted substrates have been successful for Morchella rufobrunnea. We used DNA amplicon sequencing of the Internal Transcribed Spacer (ITS) ribosomal DNA and 16S rRNA gene to follow bacterial and fungal communities in substrates during indoor morel cultivation. Our goal was to determine changes in microbial communities at key stages of morel cultivation, which included primordia development, fundament initiation, differentiation and maturation. Additionally, we compared microbial communities between trays that successfully fruited to those that produced conidia and primordia but aborted before ascocarp formation (non-fruiting). The prokaryotic community was dominated by Firmicutes belonging to Bacillus and Paenibacillus with a lower abundance of Flavobacteria. At earlier stages, the fungal community was dominated by Pezizomycetes including Morchella and other species, whereas, later in the cropping cycle Sordariomycetes dominated. Additionally, differences were observed between trays with successful fruiting, which were dominated by Gilmaniella; compared to trays that did not fruit, which were dominated by Cephalotrichum. Our findings inform understanding of microbial community dynamics during morel cultivation, and show that fungal genera, such as Gilmaniella, and prokaryotic genera, such as Bacillus, are abundant in substrates that support M. rufobrunnea fruiting.


Public health opportunities and challenges in the provision of partner notification services: the New England experience.
Partner notification services (PNS) are recommended by the Centers for Disease Control and Prevention as a public health intervention for addressing the spread of HIV and other sexually transmitted diseases (STDs). Barriers and facilitators to the partner notification process from a public health perspective have not been well described. In 2015, a coalition of New England public health STD directors and investigators formed to address the increasing STD prevalence across the region (Connecticut, Maine, Massachusetts, New Hampshire, Rhode Island, and Vermont) and to promote communication between state STD programs. To evaluate barriers and facilitators of PNS programs, a survey was administered to representatives from each state to describe PNS processes and approaches. Of the six PNS programs, Connecticut, Maine, Massachusetts, Vermont, and New Hampshire had combined HIV and STD PNS programs; Rhode Island's programs were integrated but employed separate disease intervention specialists (DIS). All states performed PNS for HIV and syphilis. Maine, New Hampshire and Vermont performed services for all gonorrhea cases. Rhode Island, Connecticut, and Massachusetts performed limited partner notification for gonorrhea due to lack of resources. None of the six states routinely provided services for chlamydia, though Maine and Vermont did so for high-priority populations such as HIV co-infected or pregnant individuals. Across all programs, clients received risk reduction counseling and general STD education as a component of PNS, in addition to referrals for HIV/STD care at locations ranging from Planned Parenthood to community- or hospital-based clinics. Notable barriers to successful partner notification across all states included anonymous partners and index cases who did not feel comfortable sharing partners' names with DIS. Other common barriers included insufficient staff, inability of DIS to identify and contact partners, and index cases declining to speak with DIS staff. In New England, state health departments use different strategies to implement PNS programs and referral to STD care. Despite this, similar challenges exist across settings, including difficulty with anonymous partners and limited state resources.


The bond length and bond energy of gaseous CrW.
Supersonically cooled CrW was studied using resonant two-photon ionization spectroscopy. The vibronically resolved spectrum was recorded over the region 21 100 to 23 400 cm(-1), showing a very large number of bands. Seventeen of these bands, across three different isotopologues, were rotationally resolved and analyzed. All were found to arise from the ground (1)Σ(+) state of the molecule and to terminate on states with Ω' = 0. The average r0 bond length across the three isotopic forms was determined to be 1.8814(4) Å. A predissociation threshold was observed in this dense manifold of vibronic states at 23 127(10) cm(-1), indicating a bond dissociation energy of D0(CrW) = 2.867(1) eV. Using the multiple bonding radius determined for atomic Cr in previous work, the multiple bonding radius for tungsten was calculated to be 1.037 Å. Comparisons are made between CrW and the previously investigated group 6 diatomic metals, Cr2, CrMo, and Mo2, and to previous computational studies of this molecule. It is also found that the accurately known bond dissociation energies of group 5/6 metal diatomics Cr2, V2, CrW, NbCr, VNb, Mo2, and Nb2 display a qualitative linear dependence on the sum of the d-orbital radial expectation values, r; this relationship allows the bond dissociation energies of other molecules of this type to be estimated.


Sauvagine cross-links to the second extracellular loop of the corticotropin-releasing factor type 1 receptor.
Contact sites between the corticotropin-releasing factor receptor type 1 (CRFR1), the sauvagine (SVG) radioligands [Tyr(0),Gln(1)]SVG ((125)I-YQS) and [Tyr(0),Gln(1), Leu(17)]SVG ((125)I-YQLS) were examined. (125)I-YQLS or (125)I-YQS was cross-linked to CRFR1 using the chemical cross-linker, disuccinimidyl suberate (DSS), which cross-links the epsilon amino groups of lysine residues that have a molecular distance of 11.4 A. DSS specifically and efficiently cross-linked (125)I-YQLS and (125)I-YQS to CRFR1. CRFR1 contains 5 putative extracellular lysine residues (Lys(110), Lys(111), Lys(113), Lys(257), and Lys(262)) that can cross-link to the 4 lysine residues (Lys(16), Lys(22), Lys(25), and Lys(27)) of the radioligands. Identification of the CNBr-cleaved fragments of CRFR1 cross-linked to (125)I-YQLS or (125)I-YQS established that the second extracellular loop of CRFR1 cross-links to Lys(16) of YQS. Additionally, site-directed mutagenesis (changing Lys to Arg in CRFR1 individually and in combination) revealed that Lys(257) in the second extracellular loop of CRFR1 is an important cross-linking site. In conclusion, it was shown that in SVG-bound CRFR1, Lys(257) of CRFR1 lies in close proximity (11.4 A) to Lys(16) of SVG.


Microbiota Perturbation or Elimination Can Inhibit Normal Development and Elicit a Starvation-Like Response in an Omnivorous Model Invertebrate.
Omnivorous animals, including humans, harbor diverse, species-rich gut communities that impact their growth, development, and homeostasis. Model invertebrates are broadly accessible experimental platforms that enable linking specific species or species groups to host phenotypes, yet often their specialized diets and distinct gut microbiota make them less comparable to human and other mammalian and gut communities. The omnivorous cockroach Periplaneta americana harbors ∼4 × 102 bacterial genera within its digestive tract and is enriched with taxa commonly found in omnivorous mammals (i.e., Proteobacteria, Bacteroidetes, and Firmicutes). These features make P. americana a valuable platform for identifying microbe-mediated host phenotypes with potential translations to mammals. Rearing P. americana insects under germfree conditions resulted in prolonging development time by ∼30% and an up to ∼8% reduction in body size along three dimensions. Germfree rearing resulted in downregulation of gene networks involved in growth, energy homeostasis, and nutrient availability. Reintroduction of a defined microbiota comprised of a subset of P. americana commensals to germfree insects did not recover normal growth and developmental phenotypes or transcriptional profiles observed in conventionally reared insects. These results are in contrast with specialist-feeding model insects (e.g., Drosophila), where introduction of a single endemic bacterial species to germfree condition-reared specimens recovered normal host phenotypes. These data suggest that understanding microbe-mediated host outcomes in animals with species-rich communities should include models that typically maintain similarly diverse microbiomes. The dramatic transcriptional, developmental, and morphological phenotypes linked to gut microbiome status in this study illustrates how microbes are key players in animal growth and evolution. IMPORTANCE Broadly accessible model organisms are essential for illustrating how microbes are engaged in the growth, development, and evolution of animals. We report that germfree rearing of omnivorous Periplaneta americana cockroaches resulted in growth defects and severely disrupted gene networks that regulate development, which highlights the importance of gut microbiota in these host processes. Absence of gut microbiota elicited a starvation-like transcriptional response in which growth and development were inhibited while nutrient scavenging was enhanced. Additionally, reintroduction of a subset of cockroach gut bacterial commensals did not broadly recover normal expression patterns, illustrating that a particular microbiome composition may be necessary for normal host development. Invertebrate microbiota model systems that enable disentangling complex, species-rich communities are essential for linking microbial taxa to specific host phenotypes.


Hypoxia enhances indoleamine 2,3-dioxygenase production in dendritic cells.
Hypoxia-associated metabolic reprogramming modulates the biological functions of many immune and non-immune cells, and affects immune response types and intensities. Adenosine and indoleamine 2,3-dioxygenase (IDO) are known immunosuppressors, and adenosine is a hypoxia-associated product. We investigated the impact of hypoxia on IDO production in dendritic cells (DCs). We found that hypoxia (1% O2) enhances IDO production in DCs, and this increase was dependent on the adenosine A3 receptor (A3R), but not A2aR or A2bR. A3R blockade during hypoxia inhibited IDO production in DCs, while A2bR blockade further enhanced IDO production. Activating A2aR had no effect on IDO production. Hypoxia (1% O2) upregulated CD86, CD274, HLA-DR, and CD54, and downregulated CD40 and CD83 in DCs as compared to normoxia (21% O2). IDO inhibition in hypoxia-conditioned DCs reversed MHC-II, CD86, CD54, and CD274 upregulation, but further downregulated CD40 and CD83. Our findings offer guidance for pharmacological administration of adenosine receptor agonists or antagonists with the goal of achieving immune tolerance or controlling insulin resistance and other metabolic disorders via IDO modulation.


Joint action of t-retinoic and valproic acids on Xenopus embryo development.
The joint action of all-trans retinoic acid (RA) with valproic acid (VPA) was examined at malformation-inducing concentrations for Xenopus embryos. The compounds were selected for testing to evaluate malformation as an endpoint for the developmental toxicity of chemical mixtures and to help assess whether joint action types can be related to modes or mechanisms of chemical toxicity for Xenopus. Three mixtures (3:1, 1:1, and 1:3 RA:VPA) were tested in 96 hr static-renewal exposures. Three separate tests were conducted on each mixture. Positive controls were also tested (i.e., 1:0 and 0:1 solutions). Using toxic unit analysis, the joint action for induction of malformations of all types, craniofacial malformations, and microcephaly was response addition, indicating the chemicals are dissimilar and non-interactive. The results support recent joint action studies in providing evidence that similar acting chemicals might be defined as chemicals with the same biochemical/molecular mechanism of action in inducing malformations in Xenopus embryos.


[A case of chronic myeloid leukemia following TS-1 therapy for advanced gastric cancer].
A 55-year-old woman had received total gastrectomy for advanced gastric cancer in March 2002, and was subsequently treated with adjuvant chemotherapy using oral anti-metabolite TS-1 for 21 months. She was well with no evidence of recurrence of gastric cancer, but leukocytosis was found in June 2005. The analysis of bone marrow revealed that Philadelphia (Ph) chromosome and bcr-abl fusion gene were positive. On the basis of these findings, the chronic phase of secondary chronic myeloid leukemia (CML) was diagnosed. Three months after being started on imatinib therapy, Ph chromosome positive cells disappeared in the bone marrow, and a complete cytogenetic response was achieved. Although CML is rare in secondary leukemia, this is, to our knowledge, the first reported case of therapy-related CML following TS-1 treatment. The present case suggested that imatinib therapy was also effective for secondary CML as well as de novo CML.


The superior laryngeal nerve.
Injury to the internal (sensory) branch of the superior laryngeal nerve during thyroidectomy is unlikely. Signs of injury are loss of sensation in the upper larynx, resulting in choking and aspiration of swallowed fluids. Injury to the external (motor) branch of the superior laryngeal nerve produces no problem of respiration, but may result in changes in the quality of the voice or even voicelessness. Injury to the external branch can be avoided if the anatomical variations are kept in mind during ligation of upper pole vessels.


[Effect of the microclimate of salt mines on T- and B-lymphocyte function in bronchial asthma patients].
A study of 55 patients with bronchial asthma revealed a reduction of the relative number of T-lymphocytes and their IgA content. The changes were most pronounced in patients during the phase of unstable remission of the disease. Disorders of the T- and B-system of lymphocytes were associated with an increased number of cortisol-resistant lymphocytes. After a course of speleotherapy one could observe an increase of the number of T-lymphocytes and their functional activity, normalization of the number of B-lymphocytes, increase of the level of IgA, reduction of the IgM content and of the relative content of cortisol-resistant lymphocytes fraction.


Impact of silver, gold, and iron oxide nanoparticles on cellular response to tumor necrosis factor.
Metallic nanomaterials are utilized in an increasing number of applications in medicine and industry. Their general toxicity was tested in numerous reports both in vitro and in vivo but limited data exist on how nanomaterials affect the activity of cellular signaling pathways activated by growth factors and cytokines. The aim of the present work was to test the hypothesis predicting that silver, gold and superparamagnetic iron oxide nanoparticles may interfere with cellular signaling activated by tumor necrosis factor (TNF) and change the final cellular outcome of TNF action. Such interference may result in disruption of homeostasis and contribute to the development of malignancies such as cancer or autoimmune diseases. Experiments were performed on HepG2 and A549 cell lines. We did not observe any interaction between nanoparticles and TNF at the level of clonogenic growth, apoptosis/necrosis induction or cell cycle. At all these endpoints, the effects of TNF and nanoparticles were additive. In contrast, gene expression analysis revealed synergistic effects. A group of genes was significantly affected only by simultaneous treatment with TNF and nanoparticles and not by any of the factors alone. Observed synergistic effect on IL10 and IL8 expression seems to be of particular importance since these cytokines are often expressed by tumor cells to inhibit tumor-targeted immune response. The observed synergistic effects of TNF and nanoparticles on cytokines expression may have significant consequences for tissue homeostasis and tumor promotion and therefore should be taken into account during development of new nanoparticle-based anticancer therapies.


Characterization of the lipolytic activity of endothelial lipase.
Endothelial lipase (EL) is a new member of the triglyceride lipase gene family previously reported to have phospholipase activity. Using radiolabeled lipid substrates, we characterized the lipolytic activity of this enzyme in comparison to lipoprotein lipase (LPL) and hepatic lipase (HL) using conditioned medium from cells infected with recombinant adenoviruses encoding each of the enzymes. In the absence of serum, EL had clearly detectable triglyceride lipase activity. Both the triglyceride lipase and phospholipase activities of EL were inhibited in a dose-dependent fashion by the addition of serum. The ratio of triglyceride lipase to phospholipase activity of EL was 0.65, compared with ratios of 24.1 for HL and 139.9 for LPL, placing EL at the opposite end of the lipolytic spectrum from LPL. Neither lipase activity of EL was influenced by the addition of apolipoprotein C-II (apoC-II), indicating that EL, like HL, does not require apoC-II for activation. Like LPL but not HL, both lipase activities of EL were inhibited by 1 M NaCl. The relative ability of EL, versus HL and LPL, to hydrolyze lipids in isolated lipoprotein fractions was also examined using generation of FFAs as an end point. As expected, based on the relative triglyceride lipase activities of the three enzymes, the triglyceride-rich lipoproteins, chylomicrons, VLDL, and IDL, were efficiently hydrolyzed by LPL and HL. EL hydrolyzed HDL more efficiently than the other lipoprotein fractions, and LDL was a poor substrate for all of the enzymes.


A systematic review of patient-reported outcome instruments measuring sleep dysfunction in adults.
Sleep dysfunction can manifest in several ways, ranging from insomnia to somnolence, and from disrupted sleep to lack of restful sleep. Measuring sleep dysfunction is an area of active research and there exist a number of patient-reported outcome instruments that measure various aspects of sleep dysfunction. However, these instruments have not been evaluated systematically. We used a conceptual model of sleep that included four physical domains of general interest to patients and investigators, and cover the breadth of this disorder: sleep initiation; sleep maintenance; sleep adequacy; and somnolence. We next considered the additional health-related quality-of-life (HR-QOL) domains of psychological and social functioning, progressing along the continuum to include health perceptions and opportunity. We then conducted a literature review to identify instruments and, using criteria developed by the Medical Outcomes Trust Scientific Advisory Committee, evaluated these instruments for their potential use in measuring sleep dysfunction. Twenty-two instruments were identified. Six instruments were found to include the four physical domains defined a priori (Basic Nordic Sleep Questionnaire, Leeds Sleep Evaluation Questionnaire, Medical Outcomes Study - Sleep Problems Measures, Pittsburgh Sleep Diary, Pittsburgh Sleep Quality Index, Self-Rated Sleep Questionnaire and the Sleep Dissatisfaction Questionnaire). Several additional instruments addressed at least some of the domains and thus may be useful for specific purposes. A few instruments addressed overall HR-QOL, but did not include all four domains of interest (Functional Outcomes of Sleep Questionnaire, Quality of Life in Insomniacs and the Sleep-Wake Activity Inventory). Two instruments had undergone extensive psychometric evaluation (Medical Outcomes Study - Sleep Problems Measures and Pittsburgh Sleep Quality Index), with only the latter reporting information about interpretability. Our review indicates that measuring sleep dysfunction in adults is an area of active research and that much work still needs to be completed, specifically the study of interpretability and the application of patient preferences or item response theory. The specific research focus should dictate instrument selection.


Comparison of transcranial brain stimulation approaches: prefrontal theta alternating current stimulation enhances working memory performance.
One of the most important cognitive functions in our everyday life is the working memory (WM). In several neuropsychiatric diseases such as ADHD or schizophrenia WM deficits can be observed, making it an attractive target for non-invasive brain stimulation methods like transcranial electrical stimulation (tES). However, the literature shows rather heterogeneous results of tES effects on WM performance. fMRI meta-analyses have identified a WM network including frontoparietal brain areas such as the dorsolateral prefrontal cortex (DLPFC) and the posterior parietal cortex (PPC). Neurophysiological studies revealed oscillatory activity in the theta band frequency range to be of crucial functional relevance for WM processes. Based on this, transcranial alternating current stimulation (tACS) in the theta frequency range targeting DLPFC and PPC in a spatially optimized way might further improve effects of tES on WM performance. Sixteen healthy subjects were stimulated with varying stimulation settings on four different days in a counterbalanced within-subject design. These setups included the application of (1) tACS with a frequency of 5 Hz (theta frequency range) over the left DLPFC and (2) the right superior parietal cortex, (3) transcranial direct current stimulation (tDCS) of the DLPFC and (4) a sham stimulation condition during the online performance of a visual delayed-match-to-sample task with varying working memory load. We introduce a procedure to calculate an optimal tES model revealing optimized high-density setups for the present study for 3 cathodes and 1 anode and stimulation currents of 1.5 mA. A significant interaction effect of stimulation type and load condition on working memory capacity was found. This was reflected by a significant improvement of WM performance in the high load condition during tACS over the left DLPFC compared with sham stimulation, which was not the case for our parietal tACS or tDCS setup. Working memory performance can be improved with optimized high-definition tACS with a frequency of 5 Hz over the left DLPFC. The conception of different mechanisms underlying transcranial electrical stimulation with alternating and direct currents is supported by these results. Patients suffering from working memory impairments due to neuropsychiatric diseases might potentially benefit from this brain stimulation approach.


SETDB1 Inhibits p53-Mediated Apoptosis and Is Required for Formation of Pancreatic Ductal Adenocarcinomas in Mice.
SETDB1, a histone methyltransferase that trimethylates histone H3 on lysine 9, promotes development of several tumor types. We investigated whether SETDB1 contributes to development of pancreatic ductal adenocarcinoma (PDAC). We performed studies with Ptf1aCre; KrasG12D; Setdb1f/f, Ptf1aCre; KrasG12D; Trp53f/+; Setdb1f/f, and Ptf1aCre; KrasG12D; Trp53f/f; Setdb1f/f mice to investigate the effects of disruption of Setdb1 in mice with activated KRAS-induced pancreatic tumorigenesis, with heterozygous or homozygous disruption of Trp53. We performed microarray analyses of whole-pancreas tissues from Ptf1aCre; KrasG12D; Setdb1f/f, and Ptf1aCre; KrasG12D mice and compared their gene expression patterns. Chromatin immunoprecipitation assays were performed using acinar cells isolated from pancreata with and without disruption of Setdb1. We used human PDAC cells for SETDB1 knockdown and inhibitor experiments. Loss of SETDB1 from pancreas accelerated formation of premalignant lesions in mice with pancreata that express activated KRAS. Microarray analysis revealed up-regulated expression of genes in the apoptotic pathway and genes regulated by p53 in SETDB1-deficient pancreata. Deletion of Setdb1 from pancreas prevented formation of PDACs, concomitant with increased apoptosis and up-regulated expression of Trp53 in mice heterozygous for disruption of Trp53. In contrast, pancreata of mice with homozygous disruption of Trp53 had no increased apoptosis, and PDACs developed. Chromatin immunoprecipitation revealed that SETDB1 bound to the Trp53 promoter to regulate its expression. Expression of an inactivated form of SETDB1 in human PDAC cells with wild-type TP53 resulted in TP53-induced apoptosis. We found that the histone methyltransferase SETDB1 is required for development of PDACs, induced by activated KRAS, in mice. SETDB1 inhibits apoptosis by regulating expression of p53. SETDB1 might be a therapeutic target for PDACs that retain p53 function.


BRS Violeta (BRS Rúbea × IAC 1398-21) grape juice powder produced by foam mat drying. Part I: Effect of drying temperature on phenolic compounds and antioxidant activity.
The BRS Violeta grape presents pulp and skin with high content of phenolic compounds (PCs) and intense purplish color. It was used as raw material for the production of juice and dehydrated products using foam mat drying at 60, 70 and 80 °C and freeze drying (control). HLPC-DAD-ESI-MSn allowed the evaluation of the quantitative and qualitative changes of the main PCs (anthocyanins, flavonols and hydroxycinnamic acid derivatives (HCAD)) present in the grapes during the processing. The use of the steam extraction method to obtain grape juice allowed a greater extraction of flavonols and, mainly derivatives of hydroxycinnamic acids, when compared with anthocyanins. Drying at 70 °C was the most suitable for the preservation of the PCs and, at the same time, for the reduction of the processing time. The powdered products presented considerable concentrations of total PCs (3-5 mg/g) and antioxidant activity (32-79 (DPPH) or 17-27 (FRAP) mg/g).


Clinical and ocular motor analysis of the infantile nystagmus syndrome in the first 6 months of life.
The infantile nystagmus syndrome (INS) usually begins in infancy and may or may not be associated with visual sensory system abnormalities. Little is known about its specific waveforms in the first 6 months of life or their relation to the developing visual system. This study identifies the clinical and ocular motility characteristics of the INS and establishes the range of waveforms present in the first 6 months of life. 27 infants with involuntary ocular oscillations typical of INS are included in this analysis. They were evaluated both clinically and with motility recordings. Eye movement analysis was performed off line from computer analysis of digitised data. Variables analysed included age, sex, vision, ocular abnormalities, head position, and null zone, neutral zone characteristics, symmetry, conjugacy, waveforms, frequencies, and foveation times. Ages ranged from 3 to 6.5 months (average 4.9 months). 15 patients (56%) had abnormal vision for age, nine (33%) had strabismus, five (19%) had an anomalous head posture, 13 (48%) had oculographic null and neutral positions, nine (33%) had binocular asymmetry, and only two showed consistent dysconjugacy. Average binocular frequency was 3.3 Hz, monocular frequency 6.6 Hz. Average foveation periods were longer and more "jerk" wave forms were observed in those patients with normal vision. Common clinical characteristics and eye movement waveforms of INS begin in the first few months of infancy and waveform analysis at this time may help with both diagnosis and visual status.


Effects of SO4(2-) or SiO3(2-) doping on the photoluminescence of NaEu(MoO4)2 nanophosphors for light-emitting diodes.
A series of red nanophosphors including NaEu(MoO4)2-x(SO4)x as well as NaEu(MoO4)2-x(SiO3)x (x = 0, 0.10, 0.20, 0.30, 0.40) were synthesized by hydrothermal and subsequent heat-treatment processes. The as-prepared pure and anion (SO4(2-) or SiO3(2-))-doped NaEu(MoO4)2 phosphors were indexed as a scheelite structure, which was revealed by X-ray diffraction data. The morphologies of SO4(2-)-doped NaEu(MoO4)2 powders were almost regularly spherical, and their grain sizes ranged from 300 to 400 nm. However, the grains of SiO3(2-)-doped phosphors appeared to be seriously inordinate and unconsolidated, as indicated by their scanning electron microscopy studies. All of the samples were excited with near-UV 395 nm light and showed the characteristic red emission of the Eu3+ 5D0 --> 7F2 transition. With the incorporation of SO4(2-) into the host, the red emission intensities of Eu3+ increased obviously and reached a maximum at x = 0.30. Nevertheless, the intensities of Eu3+ emission reduced dramatically after the SiO3(2-) doping. The optimized NaEu(MoO4)1.70(SO4)0.30 nanophosphor powders, with well-distributed fine grains and strong emission intensity, may be applied in the fabrication of phosphor-converted light-emitting diodes (LEDs).


The nutritional care of people living with dementia at home: A scoping review.
There are an increasing number of people with dementia living in their own home for longer, often supported by a family member. The symptoms of dementia can affect an individual's nutritional status, which can lead to a reduced quality of life for the person with dementia and their family members. A scoping review was conducted from July 2016 until September 2016, using a recognised framework, to explore what is currently known, and identify any gaps in the research regarding the nutritional care of people living with dementia at home. This included any interventions that may have been trialled or implemented, and the views of those living with dementia, carers and clinicians. Six electronic databases were searched from inception to July 2016. A review team was involved in screening and data extraction for selected articles. Published qualitative and quantitative studies were included that explored the nutritional care of people living with dementia at home. Methods included data extraction and conventional content analysis. Stakeholders were involved in the development of final categories. Following screening, 61 studies reported in 63 articles were included. Most studies were cross-sectional (n = 24), cohort (n = 15) or qualitative (n = 9). Only three were randomised controlled trials. Three overarching categories represented the results: Timely identification of nutritional risk and subsequent regular monitoring of nutritional status, multi-component tailored interventions and the influence of the care-giving dyad on nutritional status. Many studies identify people living at home with dementia as a vulnerable group prone to malnutrition; however, a lack of interventions exists to address the increased risk. There is a lack of research exploring the role of home care providers and healthcare professionals in the provision of nutritional care. Further research is required to explore how the emotional aspect of the care-giving dyad influences nutritional care.


Environmental temperature regulates transcription of a virulence pili operon in E. coli.
The expression in Escherichia coli K-12 of a pilus-adhesion determinant, obtained through molecular cloning from a pyelonephritic E. coli isolate, was studied at different temperatures. Strain HB101 carrying the recombinant plasmid pRHU845 agglutinated human erythrocytes after growth at 37 degrees C but not after growth at 22 degrees C. Quantitation of pilus subunit protein by an enzyme-linked immunosorbent assay (ELISA) for pilus antigen showed that synthesis of the pilus subunits was reduced at least 20-fold at 22 degrees C as compared with 37 degrees C. The 5' end of the pilus subunit structural gene, papA, was fused to the lacZ gene such that expression could be monitored at both translational and transcriptional levels. Measurements of beta-galactosidase production by the papA-lacZ hybrids provided evidence for thermoregulation of papA gene transcription. A regulatory determinant was localized to a 2-kb EcoRI-HindIII fragment encoding the papB gene and part of papA although none of the presently known pap gene products seem to be directly involved in a thermoregulatory mechanism. Comparison with other thermoregulatory systems in E. coli suggests that pap gene expression is regulated by a novel mechanism.


Molecular dynamics simulations of apo and holo forms of fatty acid binding protein 5 and cellular retinoic acid binding protein II reveal highly mobile protein, retinoic acid ligand, and water molecules.
Structural and dynamic properties from a series of 300 ns molecular dynamics, MD, simulations of two intracellular lipid binding proteins, iLBPs, (Fatty Acid Binding Protein 5, FABP5, and Cellular Retinoic Acid Binding Protein II, CRABP-II) in both the apo form and when bound with retinoic acid reveal a high degree of protein and ligand flexibility. The ratio of FABP5 to CRABP-II in a cell may determine whether it undergoes natural apoptosis or unrestricted cell growth in the presence of retinoic acid. As a result, FABP5 is a promising target for cancer therapy. The MD simulations presented here reveal distinct differences in the two proteins and provide insight into the binding mechanism. CRABP-II is a much larger, more flexible protein that closes upon ligand binding, where FABP5 transitions to an open state in the holo form. The traditional understanding obtained from crystal structures of the gap between two β-sheets of the β-barrel common to iLBPs and the α-helix cap that forms the portal to the binding pocket is insufficient for describing protein conformation (open vs. closed) or ligand entry and exit. When the high degree of mobility between multiple conformations of both the ligand and protein are examined via MD simulation, a new mode of ligand motion that improves understanding of binding dynamics is revealed.


Multi-electrode array recordings of human epileptic postoperative cortical tissue.
Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which disrupt normal brain function. Despite treatment with currently available antiepileptic drugs targeting neuronal functions, one third of patients with epilepsy are pharmacoresistant. In this condition, surgical resection of the brain area generating seizures remains the only alternative treatment. Studying human epileptic tissues has contributed to understand new epileptogenic mechanisms during the last 10 years. Indeed, these tissues generate spontaneous interictal epileptic discharges as well as pharmacologically-induced ictal events which can be recorded with classical electrophysiology techniques. Remarkably, multi-electrode arrays (MEAs), which are microfabricated devices embedding an array of spatially arranged microelectrodes, provide the unique opportunity to simultaneously stimulate and record field potentials, as well as action potentials of multiple neurons from different areas of the tissue. Thus MEAs recordings offer an excellent approach to study the spatio-temporal patterns of spontaneous interictal and evoked seizure-like events and the mechanisms underlying seizure onset and propagation. Here we describe how to prepare human cortical slices from surgically resected tissue and to record with MEAs interictal and ictal-like events ex vivo.


Perioperative blood transfusion and gastric cancer: adverse effects or unfavourable conditions of pretreatment?
The use of perioperative blood transfusion (PBT), the immunological status pre-operatively and at discharge from hospital, and the clinical course were examined retrospectively in 124 patients who underwent 'curative' resection for gastric cancer at Shinkokura Hospital, Japan from 1979 to 1988. The general condition of patients with PBT was worse than that of those without PBT and the pre-operative immunological status of patients with PBT was less favourable than that of those without PBT. At the time of discharge from hospital the immunological condition remained worse for patients who had been given PBT. The clinical course of patients with PBT was significantly worse. A dose-response relationship was evident but the types of blood products did not influence the outcome. Cox regression analysis adjusting for potentially confounding prognostic factors revealed that the clinical course was not altered by perioperative blood transfusion itself. These observations do not support the idea of adverse effects of perioperative blood transfusion on outcome of patients undergoing 'curative' resection for gastric cancer.


Evaluation of pulpal anesthesia and injection pain using IANB with pre-heated, buffered and conventional 2% lignocaine in teeth with symptomatic irreversible pulpitis-a randomized clinical study.
The efficacy of 2% lignocaine is reduced in a hot tooth. Local aesthetic agents can be preheated and buffered to increase their effectiveness. The present investigation was carried out due to limited information concerning adult patients with symptomatic irreversible pulpitis in mandibular teeth. A total of 252 individuals were included in the clinical trial in accordance with the selection criteria only after clinical study was registered with the Clinical Trial Registry of India (CTRI/2020/09/027796). Scores on the visual analog scale (VAS) and electric pulp test (EPT) on a 1-10 scale were recorded prior to the commencement of therapy. In this double-blinded study, patients were randomly divided by a co-investigator using computer randomisation (www.randomizer.org) into three groups, group A: inferior alveolar nerve blocks (IANB) with 2% lignocaine preheated at 42 °C (injected at 37 °C) (N = 84), group B: IANB of 2% lignocaine buffered with 0.18 ml of 8.4% sodium bicarbonate (N = 80) and group C: 2% lignocaine (N = 88). Excluding the dropouts of individuals (n = 11), wherein the anaesthesia failed, a total of 241 people were finally assessed 15 minutes after profound anaesthesia, endodontic access, and intraoperative pain were quantified using VAS. Pain on injection for all three groups was recorded immediately after IANB with VAS. The analysis was performed using one way ANOVA with Tukey's post hoc test and Paired T-Test using SPSS version 21. Preheated, Buffered, and conventional 2% lignocaine showed statistically significant reduction in intraoperative pain (P < 0.001) compared to pre-operative but on inter-group comparison preheated and buffered showed highly significant pain reduction compared with conventional 2% lignocaine (P < 0.001). Warm and buffered local anaesthetic (LA) were effective in reducing intraoperative discomfort than conventional LA. Preheated local anesthetics caused the least pain, followed by buffered local anesthetics, while conventional local anesthetics caused the most pain.


Comparison between blood hemoglobin concentration determined by point-of-care device and complete blood count in adult patients with dengue.
Hematocrit measurement has been an indispensable tool for monitoring plasma leakage and bleeding in dengue patients. However, hematocrit measurement by automated methods is hampered by frequent venipunctures. Utility of point-of-care hemoglobin (POC-Hb) test for monitoring dengue patients has not been established. We evaluated the relationship between hemoglobin measured by POC-Hb testing and hematocrit measured by the automated method in adult dengue patients. Adult dengue patients were recruited at two university hospitals in Thailand from October 2019 to December 2020. POC-Hb test was performed using capillary blood simultaneously with venipuncture to obtain whole blood for an automated complete blood count (CBC) analysis. The correlation of hemoglobin and hematocrit measurement was evaluated. A total of 44 dengue patients were enrolled. Twenty-nine patients (65.9%) were female, with a median age of 31 years (interquartile range 22-41). Of the enrolled patients, 30 (68.2%), 11 (25.0%), and 3 (6.8%) were classified as dengue without warning signs, with warning signs, and severe dengue, respectively. Seven patients (15.9%) had hemoconcentration, and five patients (11.3%) had bleeding. A total of 216 pairs of POC-Hb and CBC were evaluated. A significant positive correlation was observed between hemoglobin measured by POC-Hb testing and hematocrit measured by an automated CBC (r = 0.869, p <0.001). Bland-Altman analysis between hemoglobin measured by POC-Hb testing and an automated CBC showed a bias of -0.43 (95% limit of agreement of -1.81 and 0.95). Using the cutoff of POC-Hb ≥20% as a criteria for hemoconcentration, the sensitivity and specificity of hemoconcentration detected by POC-Hb device were 71.4% and 100.0%, respectively. Hemoglobin measurement by POC-Hb testing has a strong correlation with hematocrit in adult patients with dengue fever. However, the sensitivity in detecting hemoconcentration is fair. The adjunct use of capillary POC-Hb testing can decrease the frequency of venipuncture. Further study in children is encouraged.


Choline accumulation by photoreceptor cells of the rabbit retina.
Photoreceptor cells of the rabbit retina accumulate choline from the extracellular environment by an overall process that has a high affinity for choline. These cells do not synthesize acetylcholine; instead, the choline taken up is incorporated into phosphorylcholine and eventually phospholipid. A mechanism for efficient choline accumulation is presumably concomitant to the photoreceptor cell's synthesis of large amounts of membrane for outer segment membrane renewal. Its existence in the photoreceptor cell supports previous evidence that high-affinity choline uptake is not confined to neurons that release acetylcholine, but may be present wherever large amounts of choline are required.


Pathophysiology of skin flaps raised on expanded pig skin.
The pathophysiology of skin flaps raised on nonexpanded and expanded skin was studied in pigs. Skin expansion was achieved by gradual inflation of a subcutaneously implanted silicone tissue expander with sterilized isotonic saline over a period of 4 to 5 weeks. Comparisons were made of the skin flap viability and capillary blood flow among random acute and delayed skin flaps and skin flaps constructed on skin pockets implanted with noninflated or inflated tissue expanders. The extents of skin flap viability and capillary blood flow were similar among delayed skin flaps and skin flaps constructed on skin pockets but were significantly higher than those of acute random skin flaps raised on normal skin. It was concluded that skin viability and capillary blood flow in skin flaps raised on expanded skin were not compromised and the regulation of blood flow in skin flaps raised on nonexpanded or expanded skin pockets was similar to that seen in delayed skin flaps. The mechanisms of the delay phenomenon was discussed.