A novel biotin protein required for reductive carboxylation of 2-oxoglutarate by isocitrate dehydrogenase in Hydrogenobacter thermophilus TK-6.
Isocitrate dehydrogenase was purified from Hydrogenobacter thermophilus, and the corresponding gene was cloned and sequenced. The enzyme had similar structural properties to the isocitrate dehydrogenase of Escherichia coli, but differed in its catalytic properties, such as coenzyme specificity, pH dependency and kinetic parameters. Notably, the enzyme catalysed the oxidative decarboxylation of isocitrate, but not the reductive carboxylation of 2-oxoglutarate. The carboxylation reaction required the addition of cell extract and ATP-Mg, suggesting the existence of additional carboxylation factor(s). Further analysis of the carboxylation factor(s) resulted in the purification of two polypeptides. N-terminal amino acid sequencing revealed that the two polypeptides are homologues of pyruvate carboxylase with a biotinylated subunit, but do not catalyse pyruvate carboxylation. Pyruvate carboxylase was also purified, but was not active in stimulating isocitrate dehydrogenase. Isocitrate dehydrogenase, the novel biotin protein, ATP-Mg and NADH were essential for the reductive carboxylation of 2-oxoglutarate. These observations indicate that the novel biotin protein is an ATP-dependent factor, which is involved in the reverse (carboxylating) reaction of isocitrate dehydrogenase.


Hexahedron Prism-Anchored Octahedronal CeO2: Crystal Facet-Based Homojunction Promoting Efficient Solar Fuel Synthesis.
An unprecedented, crystal facet-based CeO2 homojunction consisting of hexahedron prism-anchored octahedron with exposed prism surface of {100} facets and octahedron surface of {111} facets was fabricated through solution-based crystallographic-oriented epitaxial growth. The photocatalysis experiment reveals that growth of the prism arm on octahedron allows to activate inert CeO2 octahedron for an increase in phototocatalytic reduction of CO2 into methane. The pronounced photocatalytic performance is attributed to a synergistic effect of the following three factors: (1) band alignment of the {100} and {111} drives electrons and holes to octahedron and prism surfaces, respectively, aiming to reach the most stable energy configuration and leading to a spatial charge separation for long duration; (2) crystallographic-oriented epitaxial growth of the CeO2 hexahedron prism arm on the octahedron verified by the interfacial lattice fringe provides convenient and fast channels for the photogenerated carrier transportation between two units of homojuntion; (3) different effective mass of electrons and holes on {100} and {111} faces leads to high charge carrier mobility, more facilitating the charge separation. The proposed facet-based homojunction in this work may provide a new concept for the efficient separation and fast transfer of photoinduced charge carriers and enhancement of the photocatalytic performance.


Prevalence of Nocardia species among HIV-positive patients with suspected tuberculosis.
The objective of the study was to determine the frequency of nocardiosis in HIV-positive and HIV-negative individuals clinically suspected of having tuberculosis (TB). The study population (n = 171) were those who attended chest hospitals in Khartoum State, Sudan, between January and March 2010. The patients suffered from pulmonary infections with positive acid-fast bacilli. Blood (n = 171) and sputum (n = 171) samples were collected simultaneously. Blood samples were tested serologically for the presence of antibodies using HIV/Intensified Combination Prevention (ICP) test and sputum were cultured onto Lowenstein Jensen slants according to standard methods. Isolates showing rapid growth characteristic of Nocardiae were subcultured and subsequently identified using glucose yeast extract agar medium. All candidates in the study population (n = 171) suffered from pulmonary infections, nocardiosis was diagnosed in 4% (n = 7), HIV-positive cases were 17 (9.9%). Five Nocardia species were isolated from HIV-negative patients whereas two were from HIV-positive patients. Nocardia spp. cause pulmonary infections (4.09%) in both immunocompetent (2.92%) as well as immunocompromised (1.17%) patients who attend chest clinics in Sudan.


Establishing a data base: the first step in effective management for day-treatment programs for children and adolescents.
A simple step-by-step plan for organizing and developing a computerized data base in a psychoeducational day-treatment program for children and adolescents is described. The plan is purposefully limited and simple in conception so that it can be replicated in freestanding and/or smaller programs, those who might not have the resources available when affiliated with a hospital or university. The article stresses the need for programs to make the commitment to begin a system of data collection. A table indicating measures and collection times is included.


Differential response of psychotic and obsessive symptoms to risperidone in an adolescent.
A 13-year-old boy with psychotic, depressive, and obsessive-compulsive symptoms initially presented with auditory and visual hallucinations and a lifetime of excessive worries about contamination. Family history was significant for schizophrenia and compulsive behavior. When treated with clomipramine 100 mg daily (plasma level 85 ng/mL), obsessive-compulsive symptoms but not the hallucinations improved significantly, and racing thoughts and grandiosity developed later. Haloperidol 0.5 mg daily reduced the psychotic symptoms but was poorly tolerated, and then trifluoperazine 3 mg daily was ineffective, so clomipramine was discontinued (without worsening of OCD symptoms). Trifluoperazine in combination with lithium 1500 mg daily (0.9 meq/L), and then with the addition of carbamazepine 250 mg daily (3.7 micrograms/mL), was only partially helpful. Dose reductions in any medication led to increased psychotic symptoms within days. Trifluoperazine was then replaced by risperidone 3 mg twice daily. Within 2 weeks of starting risperidone, psychotic symptoms ceased but the patient experienced an incapacitating exacerbation of obsessive-compulsive symptoms, experiencing the most severe symptoms in his illness. Sertraline 50 mg reduced the OCD symptoms only briefly. After 5 months on risperidone, risperidone and sertraline were discontinued, and the obsessive-compulsive symptoms significantly decreased within 2 weeks. These clinical observations suggest that even when risperidone has a therapeutic antipsychotic effect, it may exacerbate obsessive-compulsive symptoms in predisposed adolescents.


Detecting Endometriosis in Adolescents: Why Not Start from Self-Report Screening Questionnaires for Adult Women?
Endometriosis in adolescent girls is often diagnosed after a long delay. This diagnostic delay can be associated with more advanced stages of endometriosis and with a higher likelihood of fertility problems at a later age. A systematic review of literature and quality assessment was performed in order to identify questionnaires that were developed to identify adult women with endometriosis. Based on these questionnaires, specific questions that had been reported to be predictive for endometriosis were selected and included in a newly composed questionnaire with the aim to identify adolescents at risk of developing endometriosis. Based on the literature, we identified 5 questionnaires developed to identify adult women with endometriosis; this questionnaire contained 6 questions that had been reported to be predictive for adult endometriosis. These questions query age of menarche, cycle duration, dysmenorrhea, pain descriptors, dyschezia and urinary symptoms and were combined into a new self-report questionnaire aimed to identify adolescents at risk to develop endometriosis. We developed a self-report questionnaire aimed to identify adolescents at risk to develop endometriosis based on questions from self-report questionnaires that have been reported to identify adult women with endometriosis.


Liquid chromatography/tandem mass spectrometric analysis of penicillamine for its pharmacokinetic evaluation in dogs.
Copper storage disease occurs in multiple dog breeds and is one of the most common causes of chronic hepatitis in this species. The disease is caused by hereditary defects in copper metabolism in conjunction with high dietary copper levels. The progressive copper accumulation leads to hepatitis, cirrhosis, and eventually death if left untreated. Copper chelators are critical in modulating the effects of this disease. It is therefore of significant practicality to understand the pharmacokinetic (PK) parameters of chelating agents, particularly since they are oftentimes quite expensive. A liquid chromatography-tandem mass spectrometric (LC/MS/MS) method was developed to measure plasma levels of one of the most common chelators, d-penicillamine. The compound was discovered to exist in two forms, monomeric and dimeric, and various chemical derivatizations were tried to force the compound into one form or the other. Eventually, the simplest approach was individual determination of penicillamine and its dimer, with summation of the two quantities. This enabled determination of canine PK parameters for penicillamine based on comparison of oral and intravenous administration of the drug, including time to maximum drug level (Tmax), concentration at maximum (Cmax), clearance (Cls) and volume of distribution (Vdss). The drug was found to exist predominantly in the dimeric form in plasma, which is incapable of chelating copper owing to lack of free sulfhydryl groups and must therefore provide a storage form of the drug in equilibrium with its monomeric form in vivo. Mechanisms are discussed for the electrospray-induced fragmentation of penicillamine as well as of its dimer.


The impact of long-distance horizontal gene transfer on prokaryotic genome size.
Horizontal gene transfer (HGT) is one of the most dominant forces molding prokaryotic gene repertoires. These repertoires can be as small as approximately 200 genes in intracellular organisms or as large as approximately 9,000 genes in large, free-living bacteria. In this article we ask what is the impact of HGT from phylogenetically distant sources, relative to the size of the gene repertoire. Using different approaches for HGT detection and focusing on both cumulative and recent evolutionary histories, we find a surprising pattern of nonlinear enrichment of long-distance transfers in large genomes. Moreover, we find a strong positive correlation between the sizes of the donor and recipient genomes. Our results also show that distant horizontal transfers are biased toward those functional groups that are enriched in large genomes, showing that the trends in functional gene content and the impact of distant transfers are interdependent. These results highlight the intimate relationship between environmental and genomic complexity in microbes and suggest that an ecological, as opposed to phylogenetic, signal in gene content gains relative importance in large-genomed bacteria.


[Lymphatic metastasis intensity of and lymphadenectomy for thoracic esophageal squamous cell carcinoma].
The intensity of lymphatic metastasis consists of lymph node metastasis number (LMN) and lymph node metastasis ratio (LMR). LMR is the ratio of positive nodes to dissected nodes. LMN and LMR are 2 important prognostic factors of esophageal cancer, and are adopted in tumor staging. This study was to assess the lymphatic metastasis intensity of thoracic esophageal squamous cell carcinoma (ESCC), and explore the influential factors and lymphadenectomy pattern. A total of 120 patients with ESCC had been operated in the Second Affiliated Hospital of Sun Yat-sen University from 1998 to 2000. The lymph nodes were dissected according to the mapping scheme of the American Thoracic Society (ATS) modified by Casson et al. A total of 2 631 lymph nodes were dissected with an average of 22 lymph nodes in each case. The cervical LMR was significantly higher in the upper thoracic ESCC than in the middle and lower thoracic ESCC (20.9% vs. 12.9% and 6.8%, P<0.05). The left gastric LMR was significantly higher in the lower thoracic ESCC than in the middle and upper thoracic ESCC (37.5% vs. 17.5% and 7.1%, P<0.05). Subcarinal metastatic lymph nodes were often found in the middle thoracic ESCC. T stage, histological differentiation, and circum wall involvement degree were correlated to lymphatic metastasis intensity (P<0.05); the length of esophageal lesion had no correlation to lymphatic metastasis intensity (P>0.05). The survival time of the patients received the right thoracic esophagectomy with 3-field lymphadenectomy (3-FL) was significantly longer than that of the patients received the left thoracic esophagectomy with 2-FL (P<0.05). During the operation on ESCC, the regions with high lymphatic metastasis intensity should be dissected. T stage, histological differentiation, and circum wall involvement degree are important influential factors of lymphatic metastasis intensity. The right thoracic esophagectomy with 3-FL is superior to the left thoracic esophagectomy with 2-FL.


Induction of proinflammatory cytokines in human macrophages by influenza A (H5N1) viruses: a mechanism for the unusual severity of human disease?
In 1997, the first documented instance of human respiratory disease and death associated with a purely avian H5N1 influenza virus resulted in an overall case-fatality rate of 33%. The biological basis for the severity of human H5N1 disease has remained unclear. We tested the hypothesis that virus-induced cytokine dysregulation has a role. We used cDNA arrays and quantitative RT-PCR to compare the profile of cytokine gene expression induced by viruses A/HK/486/97 and A/HK/483/97 (both H5N1/97) with that of human H3N2 and H1N1 viruses in human primary monocyte-derived macrophages in vitro. Secretion of tumour necrosis factor alpha (TNF alpha) from macrophages infected with the viruses was compared by ELISA. By use of naturally occurring viral reassortants and recombinant viruses generated by reverse genetic techniques, we investigated the viral genes associated with the TNF-alpha response. The H5N1/97 viruses induced much higher gene transcription of proinflammatory cytokines than did H3N2 or H1N1 viruses, particularly TNF alpha and interferon beta. The concentration of TNF-alpha protein in culture supernatants of macrophages infected with these viruses was similar to that induced by stimulation with Escherichia coli lipopolysaccharide. The non-structural (NS) gene-segment of H5N1/97 viruses contributed to the increase in TNF alpha induced by the virus. The H5N1/97 viruses are potent inducers of proinflammatory cytokines in macrophages, the most notable being TNF alpha. This characteristic may contribute to the unusual severity of human H5N1 disease.


Tailoring photonic metamaterial resonances for thermal radiation.
Selective solar absorbers generally have limited effectiveness in unconcentrated sunlight, because of reradiation losses over a broad range of wavelengths and angles. However, metamaterials offer the potential to limit radiation exchange to a proscribed range of angles and wavelengths, which has the potential to dramatically boost performance. After globally optimizing one particular class of such designs, we find thermal transfer efficiencies of 78% at temperatures over 1,000°C, with overall system energy conversion efficiencies of 37%, exceeding the Shockley-Quiesser efficiency limit of 31% for photovoltaic conversion under unconcentrated sunlight. This represents a 250% increase in efficiency and 94% decrease in selective emitter area compared to a standard, angular-insensitive selective absorber.PACS: 42.70.Qs; 81.05.Xj; 78.67.Pt; 42.79.Ek.


Genetic and environmental factors in febrile seizures: a Danish population-based twin study.
The relative importance of genetic and environmental factors in the etiology of febrile seizures was estimated using a large, unselected population-based twin sample. A total of 34,076 twins (aged 12-41 years), recruited from the Danish Twin Registry, were screened for febrile seizures by questionnaire. Information was obtained from 11,872 complete pairs. Concordance rates, odds ratios and correlations were used to assess the degree of similarity in monozygotic (MZ) and dizygotic (DZ) twins. Model fitting and estimation of heritability (proportion of the population variance attributable to genetic variation) were performed using standard biometrical methods. Significantly higher probandwise concordance rates were found for MZ compared with DZ twins (0.36 and 0.12, P < 0.01). Odds ratios and correlations showed a similar pattern. An etiological model including additive genetic effects and individual-specific environmental factors provided the best fit to the data with a heritability for febrile seizures of 70% (95% CI: 61-77%). The remaining 30% of the variation could be attributed to individual-specific environmental factors. In conclusion, this study has confirmed a major impact of genetic factors in the etiology of febrile seizures. Future studies aimed at identifying the specific genetic factors and environmental exposures involved in determining febrile seizure risk are clearly warranted.


[Surgical Treatments of Renal Artery Stenosis in Takayasu Arteritis].
Refractory hypertension and renal insufficiency caused by Takayasu arteritis with renal arteries involved are difficult to treat with medicines.Instead,surgery is often recommended for refractory renovascular hypertension with renal artery stenosis of ≥70%.Although both open surgery and endovascular surgery have been applied,the preferred surgical strategy remains undetermined.This article reviews the clinical effectiveness,long-term patency,and complications of open surgery,percutaneous balloon angioplasty,and stenting in patients with Takayasu arteritis-associated renal artery stenosis.


Cervical Spine Deformity-Part 1: Biomechanics, Radiographic Parameters, and Classification.
Cervical spine deformities can have a significant negative impact on the quality of life by causing pain, myelopathy, radiculopathy, sensorimotor deficits, as well as inability to maintain horizontal gaze in severe cases. Many different surgical options exist for operative management of cervical spine deformities. However, selecting the correct approach that ensures the optimal clinical outcome can be challenging and is often controversial. We aim to provide an overview of cervical spine deformity in a 3-part series covering topics including the biomechanics, radiographic parameters, classification, treatment algorithms, surgical techniques, clinical outcome, and complication avoidance with a review of pertinent literature.


Long-term treatment with Nandrolone Decanoate impairs mesenteric vascular relaxation in both sedentary and exercised female rats.
Nandrolone Decanoate (ND) is an Anabolic Androgenic Steroid (AAS) that under abusive regimen can lead to multiple physiological adverse effects. Studies of AAS-mediated cardiovascular (CV) alterations were mostly taken from male subjects, even though women are also susceptible to the effects of AAS and gender-specific differences in susceptibility to vascular diseases exist. Here we investigate ND-induced vascular reactivity alterations in both sedentary and exercised female rats and whether these alterations depend on endothelium-derived factors. We show that chronic exposure of female Wistar rats to ND (20mg/Kg/week for 4weeks) impaired the vascular mesenteric bed (MVB) reactivity to vasodilator (acetylcholine) agonist. The endothelium-dependent Nitric Oxide (NO) component was reduced in ND-treated rats, whereas neither the endothelium-derived hyperpolarizing factor (EDHF) component nor prostanoids were altered in the MVBs. Endothelial dysfunction observed in ND-treated rats was associated with decreased eNOS (Ser1177) and Akt (Ser473) phosphorylation sites and upregulation of iNOS and NADPH oxidase expression. Exercise training by weight lifting in water did not improve the vascular alterations induced by ND treatment. ND treatment also significantly reduced the serum levels of estradiol in females, overriding its CV protective effect. These results help uncover the role of ND modulating endothelial function in the setting of CV disease caused by the abuse of AAS in females. If this translates to humans, young women abusing AAS can potentially lose the cardio protective effect rendered by estrogen and be more susceptible to CV alterations.


Hybrid solar cells based on P3HT and Si@MWCNT nanocomposite.
A nanocomposite material (Si@MWCNT) with a core/sheath heterostructure consisting of silicon nanoparticles (Si-NPs) covalently bonded onto a multi-walled carbon nanotube (MWCNT) is used as a new material for solar cells. The photovoltaic response of a poly (3-hexylthiophene) (P3HT)/Si@MWCNT hybrid film is studied by I-V measurements under simulated one-sun AM 1.5 illumination (100 mW cm(-2)). In comparison with simple P3HT/Si-NPs and P3HT/MWCNT devices, the power conversion efficiency of the (P3HT)/Si@MWCNT nano-hybrid device is enhanced by 3 and 20 times, respectively. Photo-physical and energy diagram investigations suggest that the improved device performance is attributed to the combination of efficient charge generation by the P3HT/Si-NPs bulk heterojunction and efficient charge collection as a result of MWCNT template directed carrier transport.


Performance of Xpert MTB/RIF in the diagnosis of tuberculous mediastinal lymphadenopathy by endobronchial ultrasound.
The Xpert (GeneXpert) MTB/RIF, an integrated polymerase chain reaction assay, has not been systematically studied in extrapulmonary and in particular mediastinal tuberculosis (TB). To investigate the performance of Xpert MTB/RIF in the diagnosis of intrathoracic nodal TB in a large tertiary urban medical center in the UK. We collected clinical, cytological, and microbiological data from two cohorts: 116 consecutive patients referred with mediastinal lymphadenopathy with detailed diagnostic information obtained, and an immediately subsequent second cohort of 52 consecutive patients with microbiologically confirmed mediastinal TB lymphadenopathy. All data were derived between January 2010 and October 2012. All patients underwent endobronchial ultrasound and transbronchial needle aspiration (TBNA). The performance of a single Xpert MTB/RIF assay alongside standard investigations, cytology, and microscopy/culture was evaluated against culture-confirmed TB. Microbiologically confirmed TB mediastinal lymphadenopathy was diagnosed in a total of 88 patients from both cohorts. Three culture-negative cases with associated caseating granulomatous inflammation on TBNA were given a probable diagnosis. A single Xpert MTB/RIF assay demonstrated overall sensitivity for culture-positive TB of 72.6% (62.3-81.0%). Xpert specificity from cohort 1 was 96.3% (89.1-99.1%). The positive predictive value was 88.9% (69.7-97.1%), negative predictive value was 86.5% (76.9-92.1%), and odds ratio was 51.3 (24.0-98.0) for correctly identifying culture-positive disease. Xpert captured all microscopy-positive cases (14 of 14) and the majority of microscopy-negative cases (48 of 71, 67.6%). Among the cases that were culture positive by TBNA, Xpert identified two-thirds of the multiple drug-resistant TB cases, leading to immediate regimen change up to 5 weeks ahead of positive cultures. The use of Xpert combined with cytology increased the sensitivity to 96.6%. Xpert MTB/RIF provides a rapid, useful, and accurate test to diagnose mediastinal nodal TB in intermediate-incidence settings. The additional use of TBNA cytology further enhances the sensitivity of Xpert. This combination can facilitate rapid risk assessment and prompt TB treatment.


Phlegmonous Psoas Muscle Infection Causing Sepsis and Death With Missing Postmortem Computed Tomography Scan Correlation.
Infection of the psoas muscle is a rare pathology, which carries a high risk of sepsis and is a potential cause of death. Classic symptoms include back pain and fever and it may be diagnosed premortem on computed tomography or magnetic resonance imaging, where abscess formation may be identified as a discrete rim enhancing and low-attenuation lesion. Infections without abscess formation, such as phlegmonous infection, may be more difficult to identify however, particularly if there is absence of other nonspecific findings, such as gas bubbles. We report a case of Staphylococcus aureus phlegmonous psoas muscle causing sepsis and death with no postmortem computed tomography scan correlation, where clinical history of back pain and an unknown source of sepsis was the only prompt for psoas dissection. This case highlights a potential postmortem computed tomography blind spot in abdominal pathology and we recommend dissecting the psoas muscle if sepsis is suspected but a definitive septic focus is unable to be identified.


TAR RNA binding properties and relative transactivation activities of human immunodeficiency virus type 1 and 2 Tat proteins.
Using gel shift assays, we found that the human immunodeficiency virus type 1 (HIV-1) Tat protein (Tat-1) bound both HIV-1 and HIV-2 TAR RNAs with similar high affinities. In contrast, the HIV-2 Tat protein (Tat-2) bound only TAR-2 RNA with high affinity. We conclude that the weak in vivo activity of Tat-2 on the HIV-1 long terminal repeat that has been observed previously is likely the result of low affinity for TAR-1 RNA. Additionally, TAR-2 RNA was found to contain multiple specific binding sites for Tat proteins. GAL4-Tat fusion proteins were analyzed to compare the relative transactivation activities of Tat-1 and Tat-2 in the absence of requirements for binding to TAR RNAs. The GAL4-Tat-2 protein was found to transactivate synthetic promoters containing GAL4 binding sites at levels severalfold higher than did the GAL4-Tat-1 protein.


Prostate cancer outcome in Burkina Faso.
African-American black men race is one of non-modifiable risk factors confirmed for prostate cancer. Many studies have been done in USA among African- American population to evaluate prostate cancer disparities. Compared to the USA very few data are available for prostate cancer in Sub-Saharan African countries. The objective of this study was to describe incident prostate cancer (PC) diagnosis characteristics in Burkina Faso (West Africa). We performed a prospective non randomized patient's cohort study of new prostate cancer cases diagnosed by histological analysis of transrectal prostate biopsies in Burkina Faso. Study participants included 166 patients recruited at the urology division of the university hospital of Ouagadougou. Age of the patients, clinical symptoms, digital rectal examination (DRE) result, serum prostate-specific antigen (PSA) level, histological characteristics and TNM classification were taking in account in this study. 166 transrectal prostate biopsies (TRPB) were performed based on high PSA level or abnormal DRE. The prostate cancer rate on those TRPB was 63, 8 % (n=106). The mean age of the patients was 71, 5 years (52 to 86). Urinary retention was the first clinical patterns of reference in our institution (55, 7 %, n = 59). Most patients, 56, 6 % (n = 60) had a serum PSA level over than 100 ng/ml. All the patients had adenocarcinoma on histological study of prostate biopsy cores. The majority of cases (54, 7 % n = 58) had Gleason score equal or higher than 7. Prostate cancer is diagnosed at later stages in our country. Very high serum PSA level and poorly differentiated tumors are the two major characteristics of PC at the time of diagnosis.


Thigh burns from exploding e-cigarette lithium ion batteries: First case series.
E-cigarette (EC) use has risen meteorically over the last decade. The majority of these devices are powered by re-chargeable lithium ion batteries, which can represent a fire hazard if damaged, over-heated, over-charged or stored inappropriately. There are currently no reports in the medical literature of lithium ion battery burns related to EC use and no guidance on the appropriate management of lithium ion battery associated injuries. We report two individual cases of burn resulting from explosion of EC re-chargeable lithium ion batteries. Both patients required in-patient surgical management. We provide evidence that lithium ion battery explosions can be associated with mixed thermal and alkali chemical burns, resulting from the significant discharge of thermal energy and the dispersal of corrosive lithium ion compounds. We would recommend, as with other elemental metal exposures, caution in exposing lithium ion battery burns to water irrigation. Early and thorough cleaning and debridement of such burns, to remove residual lithium contamination, may limit the risk of burn wound extension and potentially improve outcomes.


Phase I dose-escalation trial of iodine 131-labeled monoclonal antibody OKB7 in patients with non-Hodgkin's lymphoma.
Eighteen patients with recurrent or refractory CD21-positive, non-Hodgkin's lymphoma (NHL) were treated in a phase IA dose-escalation therapeutic trial of iodine 131 labeled to a fixed dose of OKB7. Individual doses of 30 to 50 mCi of 131I on 25 mg OKB7 were administered 2 to 3 days apart to achieve four total 131I-OKB7 dose levels of 90, 120, 160, and 200 mCi. Pharmacology, dosimetry, therapeutic effects, toxicity, human anti-mouse antibody (HAMA) response, and maximum-tolerated dose (MTD) were determined. Patients were evaluated by imaging studies (including whole-body gamma camera or single-photon emission computed tomography [SPECT] scans), flow cytometric analysis, bone marrow biopsy, and serial blood sampling. Median plasma and whole-body half-lives (T1/2) were 16 hours and 14 hours, respectively. Plasma and whole-body radiation doses were 0.0081 Gy/mCi and 0.0022 Gy/mCi, respectively. Specific tumor visualization was noted in eight of 18 patients. HAMA was detected in 12 of 16 patients. Nonhematologic toxicity was limited to asymptomatic elevations of thyroid-stimulating hormone (TSH) in five of 15 patients. Hematologic toxicity was observed in six of 18 patients, but was severe in only two patients. MTD in patients with diffuse lymphomatous bone marrow involvement was determined to be 200 mCi in four divided doses of 50 mCi 131I/25 mg OKB7. Antitumor activity was observed in 13 of 18 patients (one partial response [PR] and 12 mixed responses) and was dependent on the 131I-OKB7 dose administered. In general, palpable peripheral lymphadenopathy, enlarged spleens, skin lesions, and circulating OKB7-positive peripheral lymphocytes responded most readily to treatment. 131I-OKB7 was safely administered to a patient in leukemic phase of NHL with prompt subsequent loss of approximately 1 kg of tumor cells from the peripheral blood without associated tumor lysis syndrome. Because antitumor activity with tolerable toxicity was observed in the majority of this group of heavily pretreated patients, phase II investigation of mAb OKB7 radioconjugates in the therapy of NHL is warranted.


A case report of cardia cancer complicated with idiopathic muscular hypertrophy of the oesophagus treated with thoracoscopic surgery.
The incidence of idiopathic muscular hypertrophy of oesophagus (IMHE) is low, and <100 cases of IMHE have been reported. IMHE is a benign oesophageal disease, characterised by hyperplasia of all layers of the wall and in particular, muscle layer. Only a few cases have been reported regarding its clinical symptoms and images. In this present case, we report a cardia cancer with IMHE, showing significant hypertrophy of muscular layer of middle part of the oesophagus and successfully treated with minimally invasive thoracoscopic surgery.


Innovative approaches to inservice education.
The many challenges facing nursing educators today include meeting the regulatory agency education standards while reaching staff on a variety of shifts/schedules. This article describes a model for the development, implementation, and evaluation of required inservice education that meets this challenge. Creative teaching strategies are discussed that facilitate learning at the individual level, encourage staff participation at the unit level, and contain costs at the department level.


Molecular mechanisms of oxidative stress in airways and lungs with reference to asthma and chronic obstructive pulmonary disease.
Oxidative stress is an important pathophysiological component of airway diseases such as asthma and chronic obstructive pulmonary disease (COPD), which cause significant morbidity and mortality. Oxidative stress leads to the activation of transcription factors and signaling pathways, partly through the activation of the innate immune response through toll-like receptors 2 and 4. Such activation leads to the release of cytokines and chemokines. In addition, adaptive immune responses are initiated through activation of dendritic cells and antigen presentation to T-helper cells, with direct activation of NKT cells. Corticosteroid insensitivity is a feature of severe asthma and COPD, and oxidative stress is an important factor in its development by inhibition of HDAC-2 activity and expression through serine hyperphosphorylation. Activation of kinases such as p38 mitogen-activated protein kinase or phospho-inositol 3-kinase delta may also be involved through phosphorylation of the glucocorticoid receptor. Antioxidants may prove to be beneficial in inhibiting inflammatory responses and restoring corticosteroid function.


Mechanisms of Resistance to Macrolide Antibiotics among Staphylococcus aureus.
Methicillin resistant Staphylococcus aureus strains pose a serious treatment problem because of their multi-drug resistance (MDR). In staphylococcal strains, resistance to macrolides, lincosamides, and streptogramin B (MLSB) correlates with resistance to methicillin. The rapid transmission of erm genes responsible for MLSB resistance has strongly limited the clinical application of traditional macrolides such as erythromycin. On the other hand, in the age of increasing insensitivity to antibiotics the idea of implementing a therapy based on older generation drugs brings hope that the spread of antibiotic resistance will be limited. A thorough understanding of the resistance mechanisms contributes to design of antibiotics that avoid bacterial insensitivity. This review highlights the mechanisms of action of macrolides and mechanism of resistance to these antibiotics among Staphylococcus aureus.


The origin of aromaticity: important role of the sigma framework in benzene.
The physical nature of aromaticity is addressed at a high ab initio level. It is conclusively shown that the extrinsic aromatic stabilization energy of benzene E(ease)B, estimated relative to its linear polyene counterpart(s), is very well-reproduced at the Hartree-Fock (HF) level. This is a consequence of the fact that the contributions arising from the zero-point vibrational energy (ZPVE) and electron correlation are rather small. More specifically, they yield together 2.0 kcalmol(-1) to the destabilization of benzene. A careful scrutiny of the HF energies by virial theorem shows further that the kinetic energies of the sigma and pi electrons E(T)HF(sigma) and E(T)HF(pi) are strictly additive in the gauge linear zig-zag polyenes, which also holds for their sum Et(T)HF This finding has the important corollary that E(ease)B is little dependent on the choice of the homodesmic reactions involving zig-zag polyenes. A detailed physical analysis of the sigma- and pi-electron contributions to extrinsic aromaticity requires explicit introduction of the potential energy terms Vne, Vee, and Vnn, which signify Coulomb interactions between the electrons and the nuclei. The Vee term involves repulsive interaction Vee(sigmapi) between the sigma and pi electrons, which cannot be unequivocally resolved into sigma and pi contributions. The same holds for the Vnn energy, which implicitly depends on the electron density distribution via the Born-Oppenheimer (BO) potential energy surface. Several possibilities for partitioning Vee(sigmapi) and Vnn terms into sigma and pi components are examined. It is argued that the stockholder principle is the most realistic, which strongly indicates that E(ease)B is a result of favorable sigma-framework interactions. In contrast, the pi-electron framework prefers the open-chain linear polyenes.


Perforations of the rectosigmoid colon induced by cleansing enema: CT findings in 14 patients.
We report the computed tomographic (CT) features of colorectal perforations caused by cleansing enema. We reviewed the medical records and CT studies of all patients with colorectal perforations caused by a cleansing enema. Fourteen patients (10 men, four women; average age = 80 years) were included in the study. The most common presenting symptoms were severe abdominal pain and fever. CT was performed within 48 h after the event in most patients. Extraluminal air in the perirectal fat was the most frequent finding on CT and was present in all patients. Additional findings were extraperitoneal ( n = 9), intraperitoneal ( n = 3) and /or subcutaneous ( n = 3) air, free fluid ( n = 9), extraluminal feces ( n = 8), and focal bowel wall thickening ( n = 4). No leak of contrast from the rectum was observed in any patient including the one patient in whom contrast was administered rectally. Ten patients were treated by surgery; five recovered and the other five died. The other four patients were treated conservatively and all four died. The diagnosis of colorectal perforation can be made on CT. Because the diagnosis is not always suspected by the clinician, the radiologist may be the first to suggest it. Therefore, the radiologist should be familiar with the CT features of a potentially lethal, rectally induced perforation.


Infantile cystinosis presenting as chronic constipation.
An infant presented with persistent constipation as the first clinical symptom of cystinosis before evidence of renal Fanconi's syndrome. The history of muscle weakness, failure to thrive, polydipsia, and polyuria prompted subsequent clinical and laboratory evaluations, leading to the correct diagnosis of cystinosis. Primary and persistent constipation as an initial manifestation of cystinosis has not been reported previously. Although rare, cystinosis should be considered in the differential diagnosis of constipation if other suggestive signs or symptoms are present.


Conformational unfolding studies of three-disulfide mutants of bovine pancreatic ribonuclease A and the coupling of proline isomerization to disulfide redox reactions.
The equilibrium stability and conformational unfolding kinetics of the [C40A, C95A] and [C65S, C72S] mutants of bovine pancreatic ribonuclease A (RNase A) have been studied. These mutants are analogues of two nativelike intermediates, des[40-95] and des[65-72], whose formation is rate-limiting for oxidative folding and reductive unfolding at 25 degrees C and pH 8.0. Upon addition of guanidine hydrochloride, both mutants exhibit a fast conformational unfolding phase when monitored by absorbance and fluorescence, as well as a slow phase detected only by fluorescence which corresponds to the isomerizations of Pro93 and Pro114. The amplitudes of the slow phase indicate that the two prolines, Pro93 and Pro114, are fully cis in the folded state of the mutants and furthermore that the 40-95 disulfide bond is not responsible for the quenching of Tyr92 fluorescence observed in the slow unfolding phase, contrary to an earlier proposal [Rehage, A., and Schmid, F. X. (1982) Biochemistry 21, 1499-1505]. The ratio of the kinetic unfolding m value to the equilibrium m value indicates that the transition state for conformational unfolding in the mutants exposes little solvent-accessible area, as in the wild-type protein, indicating that the unfolding pathway is not dramatically altered by the reduction of the 40-95 or 65-72 disulfide bond. The stabilities of the folded mutants are compared to that of wild-type RNase A. These stabilities indicate that the reduction of des[40-95] to the 2S species is rate-limited by global conformational unfolding, whereas that of des[65-72] is rate-limited by local conformational unfolding. The isomerization of Pro93 may be rate-limiting for the reduction of the 40-95 disulfide bond in the native protein and in the des[65-72] intermediate.


MR features of tuberculous arachnoiditis.
Although tuberculosis is an uncommon cause of spinal arachnoiditis, it needs to be differentiated from other causes of arachnoiditis because it is a treatable disease. Myelography, which is the imaging modality of choice for the chronic adhesive stage of tuberculous arachnoiditis, usually reveals irregularity of the thecal sac, nodularity and thickening of nerve roots, clumping of roots to each other or to the thecal sac, or CSF block. Recently Gd-diethylenetriamine pentaacetic acid enhanced MRI has been found to be useful for the active phase of tuberculous infection, effectively demonstrating abnormally thickened and enhancing meninges, intra- and extramedullary tuberculous nodules, and spinal cord changes. We present the MRI features in a case of acute tuberculous arachnoiditis.


An unusual case of gender-associated mitochondrial DNA heteroplasmy: the mytilid Musculista senhousia (Mollusca Bivalvia).
Doubly Uniparental Inheritance (DUI) represents the most outstanding exception to matrilinear inheritance of mitochondrial DNA (mtDNA), typical of Metazoa. In a few bivalve mollusks, two sex-linked mtDNAs (the so-called M and F) are inherited in a peculiar way: both daughters and sons receive their F from the mother, whereas sons inherit M from the father (males do not transmit F to their progeny). This realizes a double mechanism of transmission, in which M and F mtDNAs are inherited uniparentally. DUI systems represent a unique experimental model for testing the evolutionary mechanisms that apply to mitochondrial genomes and their transmission patterns as well as to mtDNA recombination. A new case of DUI is described in Musculista senhousia (Mollusca: Bivalvia: Mytilidae). Its heteroplasmy pattern is in line with standard DUI. Sequence variability analysis evidenced two main results: F haplotypes sequence variability is higher than that of M haplotypes, and F mitochondrial haplotypes experience a higher mutation rate in males' somatic tissues than in females' ones. Phylogenetic analysis revealed also that M. senhousia M and F haplotypes cluster separately from that of the other mytilids. Sequence variability analysis evidenced some unexpected traits. The inverted variability pattern (the F being more variable than M) was new and it challenges most of the rationales proposed to account for sex-linked mtDNA evolution. We tentatively related this to the history of the Northern Adriatic populations analyzed. Moreover, F sequences evidenced a higher mutation level in male's soma, this variability being produced de novo each generation. This suggests that mechanisms evolved to protect mtDNA in females (f.i. antioxidant gene complexes) might be under relaxed selection in males. Phylogenetic analysis of sex-linked haplotypes confirmed that they have switched their roles during the evolutionary history of mytilids, at variance to what has been observed in unionids. Consequently, reciprocal monophyly of M and F lineages got easily lost because of role-reversals and consequent losses of M lineages, as already observed in Mytilus.


Association between indoxyl sulfate and bone histomorphometry in pre-dialysis chronic kidney disease patients.
Experimental studies have suggested that indoxyl sulfate (IS), a protein-bound uremic toxin, may be involved in the development of renal osteodystrophy. evaluate the association between IS levels and biochemical parameters related to mineral metabolism and bone histomorphometry in a cohort of pre-dialysis chronic kidney disease (CKD) patients. This is a post-hoc analysis of an observational study evaluating the association between coronary calcification and bone biopsy findings in 49 patients (age: 52 ± 10 years; 67% male; estimated glomerular filtration rate: 36 ± 17 ml/min). Serum levels of IS were measured. Patients at CKD stages 2 and 3 presented remarkably low bone formation rate. Patients at CKD stages 4 and 5 presented significantly higher osteoid volume, osteoblast and osteoclast surface, bone fibrosis volume and bone formation rate and a lower mineralization lag time than CKD stage 2 and 3 patients. We observed a positive association between IS levels on one hand and the bone formation rate, osteoid volume, osteoblast surface and bone fibrosis volume on the other. Multivariate regression models confirmed that the associations between IS levels and osteoblast surface and bone fibrosis volume were both independent of demographic and biochemical characteristics of the study population. A similar trend was observed for the bone formation rate. Our findings demonstrated that IS is positively associated with bone formation rate in pre-dialysis CKD patients.


A novel 40S multi-snRNP complex isolated from rat liver nuclei.
Two structurally distinct RNP complexes (MI and MII), each with a sedimentation value of approx. 40S, were isolated from rat liver nuclear extracts by sucrose gradient centrifugation and subsequent native gel electrophoresis of the 40S hnRNP-containing fractions. MII RNP contained the bulk of hnRNA and hnRNP proteins (i.e. the 32-45KD core proteins and polypeptides of 60-80 and 110-130KD). MI RNP was characterized by the exclusive presence of U-snRNAs (U1, U2, U4, U5 and U6), their well known snRNP polypeptides and a number of Sm-associated proteins in the range of 50-210KD. Immunoselection experiments employing a monoclonal antibody with an established specificity for the U2-snRNP-specific B" polypeptide proved that the RNA and protein components characteristic of MI were part of a single multi-snRNP unit. The prominent 200/210KD protein doublet of MI was identified immunochemically as the rat homologue of the yeast PRP8 protein, a known U5-associated splicing component. Based on the major biochemical and immunochemical features of MI and MII RNP complexes, we conclude that MII represents the monomeric 40S hnRNP structure, whereas MI defines a novel multi-snRNP entity.


Midbrain organoids mimic early embryonic neurodevelopment and recapitulate LRRK2-p.Gly2019Ser-associated gene expression.
Human brain organoid models that recapitulate the physiology and complexity of the human brain have a great potential for in vitro disease modeling, in particular for neurodegenerative diseases, such as Parkinson disease. In the present study, we compare single-cell RNA-sequencing data of human midbrain organoids to the developing human embryonic midbrain. We demonstrate that the in vitro model is comparable to its in vivo equivalents in terms of developmental path and cellular composition. Moreover, we investigate the potential of midbrain organoids for modeling early developmental changes in Parkinson disease. Therefore, we compare the single-cell RNA-sequencing data of healthy-individual-derived midbrain organoids to their isogenic LRRK2-p.Gly2019Ser-mutant counterparts. We show that the LRRK2 p.Gly2019Ser variant alters neurodevelopment, resulting in an untimely and incomplete differentiation with reduced cellular variability. Finally, we present four candidate genes, APP, DNAJC6, GATA3, and PTN, that might contribute to the LRRK2-p.Gly2019Ser-associated transcriptome changes that occur during early neurodevelopment.


Increase in passive stiffness at reduced airway smooth muscle length: potential impact on airway responsiveness.
The amplitude of strain in airway smooth muscle (ASM) produced by oscillatory perturbations such as tidal breathing or deep inspiration (DI) influences the force loss in the muscle and is therefore a key determinant of the bronchoprotective and bronchodilatory effects of these breathing maneuvers. The stiffness of unstimulated ASM (passive stiffness) directly influences the amplitude of strain. The nature of the passive stiffness is, however, not clear. In this study, we measured the passive stiffness of ovine ASM at different muscle lengths (relative to in situ length, which was used as a reference length, L(ref)) and states of adaptation to gain insights into the origin of this muscle property. The results showed that the passive stiffness was relatively independent of muscle length, possessing a constant plateau value over a length range from 0.62 to 1.25 L(ref). Following a halving of ASM length, passive stiffness decreased substantially (by 71%) but redeveloped over time ( approximately 30 min) at the shorter length to reach 65% of the stiffness value at L(ref), provided that the muscle was stimulated to contract at least once over a approximately 30-min period. The redevelopment and maintenance of passive stiffness were dependent on the presence of Ca(2+) but unaffected by latrunculin B, an inhibitor of actin filament polymerization. The maintenance of passive stiffness was also not affected by blocking myosin cross-bridge cycling using a myosin light chain kinase inhibitor or by blocking the Rho-Rho kinase (RhoK) pathway using a RhoK inhibitor. Our results suggest that the passive stiffness of ASM is labile and capable of redevelopment following length reduction. Redevelopment and maintenance of passive stiffness following muscle shortening could contribute to airway hyperresponsiveness by attenuating the airway wall strain induced by tidal breathing and DI.


Topology of conical/surface intersections among five low-lying electronic states of CO2: multireference configuration interaction calculations.
Multi-reference configuration interaction with single and double excitation method has been utilized to calculate the potential energy surfaces of the five low-lying electronic states (1)A1, (1)A2, (3)A2, (1)B2, and (3)B2 of carbon dioxide molecule. Topology of intersections among these five states has been fully analyzed and is associated with double-well potential energy structure for every electronic state. The analytical potential energy surfaces based on the reproducing kernel Hilbert space method have been utilized for illustrating topology of surface crossings. Double surface seam lines between (1)A1 and (3)B2 states have been found inside which the (3)B2 state is always lower in potential energy than the (1)A1 state, and thus it leads to an angle bias collision dynamics. Several conical∕surface intersections among these five low-lying states have been found to enrich dissociation pathways, and predissociation can even prefer bent-geometry channels. Especially, the dissociation of O((3)P) + CO can take place through the intersection between (3)B2 and (1)B2 states, and the intersection between (3)A2 and (1)B2 states.


Band theoretical approaches to topological physics in strongly-correlatedf-electron Kondo systems.
First-principles band structure theory on the basis of the density functional theory (DFT) plays an essential role in the investigation of topological properties of weakly-correlated systems. DFT band structures show clear bulk band crossings for Weyl and Dirac semimetals, and surface band crossings for topological insulators and topological-crystalline insulators. In contrast, for strongly-correlatedf-electron systems, their topological properties are relatively less explored because the simple DFT does not work properly in describing the electronic structures of strongly-correlatedfelectrons. In this perspective, we examine the band theoretical approaches to topological properties of strongly-correlatedf-electron Kondo systems. We recapitulate current status of understanding of electronic structures and topological properties of strongly-correlated 4f-electron systems, such as Ce, SmB6, and g-SmS, and also a 5f-electron system PuB4, the electronic structures of which were investigated by the DFT combined with the dynamical mean-field theory (DFT + DMFT). Finally, we provide future directions and perspectives of improving theoretical band approaches to search for new topologicalf-electron systems, as an outlook.


Isomeric separation in donor-acceptor systems of Pd(II) and Pt(II) and a combined structural, electrochemical and spectroelectrochemical study.
Compounds of the form [(pap)M(Q(2-))] (pap = phenylazopyridine; Q = 3,5-di-tert-butyl-benzoquinone, M = Pd, 1a and 1b, M = Pt, 2a and 2b; Q = 4-tert-butyl-benzoquinone, M = Pd, 3a and 3b; M = Pt, 4a and 4b) were synthesized in a one-pot reaction. The geometrical isomers, which are possible because of the built in asymmetry of these ligands, have been separated by using different temperatures and variable solubility. Structural characterization of 1b shows that the metal centers are in a square planar environment, the pap ligand is in the unreduced neutral state and the quinones are in the doubly reduced, Q(2-) catecholate form. Cyclic voltammetric measurements on the complexes display two one-electron oxidations and two one-electron reductions. EPR and vis-NIR spectra of the one-electron oxidized forms of the complexes indicate that the first oxidation takes place on the Q(2-) ligands to produce a metal bound semiquinone (Q˙(-)) radical. Reduction takes place on the pap ligand, generating metal bound pap˙(-) as seen from the (14)N (I = 1) coupling in their EPR spectrum. All the complexes in their [(pap)M(Q(2-))] neutral forms show strong absorptions in the NIR region which are largely LLCT (ligand to ligand charge transfer) in origin. These NIR bands can be tuned over a wide energy range by varying the metal center as well as the Q ligand. In addition, the intensity of NIR bands can be switched on and off by a simple electron transfer at relatively low potentials. DFT studies were used to corroborate these findings.


Generation of quasistatic magnetic fields.
A prototype switching system has been developed which can switch 20 kA at 230 V furnished by low-voltage dc generators at the National Magnet Laboratory. The energy is used to generate quasistatic magnetic fields in liquid-nitrogen-cooled copper coils for times approaching 1 s. The present arrangement is limited to fields up to 40 T. The silicon-controlled rectifier switching system, protective devices, circuit operation, and magnet structures are discussed.


Lack of developmental neurotoxicity of MN rgp 120/HIV-1 administered subcutaneously to neonatal rats.
The potential for neurotoxic effects was evaluated in rat off-spring after exposure in utero and/or during the neonatal period to a recombinant subunit vaccine of gp120 prepared from the MN strain of HIV-1 (MN rgp 120/HIV-1). Thirty pregnant female rats were given MN rgp120/HIV-1 with alum adjuvant, and 30 rats were given vehicle, once every 3 days from Day 1 of presumed gestation until parturition. One pup/sex/litter from treated and control group dams were given a daily subcutaneous injection, from Day 1 through Day 22 postpartum (PP) of vehicle, MN rgp120/HIV-1, MN rgp120/HIV-1 with alum, or MN rgp120/HIV-1 with QS-21 adjuvant. Neurobehavioral and physical development were evaluated (preweaning reflex and development, sexual maturation, motor activity, acoustic startle, passive avoidance, functional observational battery, and water M-maze testing), and tissues were processed for anatomical examination (whole and regional brain weights, and neuropathology). Administration of MN rgp120/HIV-1, with or without adjuvant, to pups did not cause any persistent effect on any parameter evaluated. Neurohistological examination did not reveal any pathological effects related to treatment. Thus, MN rgp120/HIV-1 alone or formulated as a vaccine does not cause neurotoxicity or developmental toxicity in neonatal rats after exposure in utero and/or during the neonatal period.


Developmental regulation of diacylglycerol acyltransferase family gene expression in tung tree tissues.
Diacylglycerol acyltransferases (DGAT) catalyze the final and rate-limiting step of triacylglycerol (TAG) biosynthesis in eukaryotic organisms. DGAT genes have been identified in numerous organisms. Multiple isoforms of DGAT are present in eukaryotes. We previously cloned DGAT1 and DGAT2 genes of tung tree (Vernicia fordii), whose novel seed TAGs are useful in a wide range of industrial applications. The objective of this study was to understand the developmental regulation of DGAT family gene expression in tung tree. To this end, we first cloned a tung tree gene encoding DGAT3, a putatively soluble form of DGAT that possesses 11 completely conserved amino acid residues shared among 27 DGAT3s from 19 plant species. Unlike DGAT1 and DGAT2 subfamilies, DGAT3 is absent from animals. We then used TaqMan and SYBR Green quantitative real-time PCR, along with northern and western blotting, to study the expression patterns of the three DGAT genes in tung tree tissues. Expression results demonstrate that 1) all three isoforms of DGAT genes are expressed in developing seeds, leaves and flowers; 2) DGAT2 is the major DGAT mRNA in tung seeds, whose expression profile is well-coordinated with the oil profile in developing tung seeds; and 3) DGAT3 is the major form of DGAT mRNA in tung leaves, flowers and immature seeds prior to active tung oil biosynthesis. These results suggest that DGAT2 is probably the major TAG biosynthetic isoform in tung seeds and that DGAT3 gene likely plays a significant role in TAG metabolism in other tissues. Therefore, DGAT2 should be a primary target for tung oil engineering in transgenic organisms.


Thermodynamics of electron transfer in oxygenic photosynthetic reaction centers: a pulsed photoacoustic study of electron transfer in photosystem I reveals a similarity to bacterial reaction centers in both volume change and entropy.
The thermodynamic properties of electron transfer in biological systems are far less known in comparison with that of their kinetics. In this paper the enthalpy and entropy of electron transfer in the purified photosystem I trimer complexes from Synechocystis sp. PCC 6803 have been studied, using pulsed time-resolved photoacoustics on the 1 micros time scale. The volume contraction of reaction centers of photosystem I, which results directly from the light-induced charge separation forming P(700+F(A)/F(B-) from the excited-state P700*, is determined to be -26 +/- 2 A3. The enthalpy of the above electron-transfer reaction is found to be -0.39 +/- 0.1 eV. Photoacoustic estimation of the quantum yield of photochemistry in the purified photosystem I trimer complex showed it to be close to unity. Taking the free energy of the above reaction as the difference of their redox potentials in situ allows us to calculate an apparent entropy change (TDeltaS) of +0.35 +/- 0.1 eV. These values of DeltaV and TDeltaS are similar to those of bacterial reaction centers. The unexpected sign of entropy of electron transfer is tentatively assigned, as in the bacterial case, to the escape of counterions from the surface of the particles. The apparent entropy change of electron transfer in biological system is significant and cannot be neglected.


Oral Cancer Screening at Workplace in India-One-year Follow-up.
Oral cancer remains the commonest form of cancer and cancer-related deaths among Indian males due to popularity of avoidable risk factors such as tobacco and alcohol use. A workplace oral cancer screening and tobacco cessation study was commenced on World No Tobacco Day 2007 at a chemical industry in rural Maharashtra. The objectives were to screen the employees for oral neoplasia and to correlate it with their tobacco consumption pattern. In addition, the objective was to provide tobacco cessation services at the workplace. This is an interventional cohort study among 104 employees of a chemical industrial unit in rural Maharashtra. Naked eye examination of the oral cavity was performed for all employees by a doctor irrespective of the tobacco habits at the beginning and at the end of 1 year. In between, the tobacco users were regularly examined during each follow-up. Through personal interviews of the participants, data were manually recorded and were transferred to electronic data base. Data analysis was conducted in STATA™ 8.2 on intention to treat basis. Among the 104 employees, 50 (48.08%) were current tobacco users at the beginning of the program. Oral precancers were seen exclusively among 20 (40%) tobacco users. After 1 year of workplace tobacco cessation intervention, 80% of oral precancers regressed. This shows that screening of the oral cavity at the workplace is effective when combined with tobacco cessation.


Noninvasive assessment of renal artery stenosis: current imaging protocols and future directions in ultrasonography.
Doppler ultrasound is one of many modalities that have been evaluated for the detection of renal artery stenosis. The lack of standardization in examination protocols and diagnostic criteria, as well as the wide differences in reported accuracy among different laboratories have, however, prevented universal acceptance of this technique as a reliable screening test for renal artery stenosis. The recent introduction of ultrasound contrast agents has substantially expanded the potential of color Doppler ultrasound. The use of microbubble echoenhancers in combination with harmonic Doppler imaging has been shown to improve diagnostic confidence by improving the operator's ability to visualize the renal arteries, and to significantly reduce the number of equivocal examinations. Moreover, contrast-enhanced Doppler ultrasound can provide functional information through analysis of renal time-intensity wash in/wash out curves. State-of-the-art contrast-enhanced Doppler ultrasound seems to have the potential to become a useful screening test for patients at risk from renovascular hypertension.


Abscisic Acid Control of Lectin Accumulation in Wheat Seedlings and Callus Cultures : Effects of Exogenous ABA and Fluridone.
Wheat germ agglutinin is found in wheat embryos and a similar lectin is present in the roots of older plants. We report here that 10 micromolar abscisic acid (ABA) produces an average two to three-fold enhancement in the amount of lectin in the shoot base and the terminal portion of the root system of hydroponically grown wheat seedlings. Although ABA stunts seedling growth, a similar growth inhibition produced by ancymidol is not accompanied by elevated lectin levels. To further clarify the role of ABA, wheat callus cultures were employed. Callus derived from immature embryos was grown on growth medium containing various combinations of ABA and 2,4-dichlorophenoxyacetic acid. Those grown in the presence of 10 micromolar ABA exhibit the largest increases in lectin compared to material grown on other regimes. The involvement of ABA in lectin accumulation was further probed with fluridone, an inhibitor of carotenoid synthesis which has also been linked to depressed levels of endogenous ABA. Wheat seedlings grown in the presence of 1 or 10 milligrams per liter fluridone have few or no carotenoids, and wheat germ agglutinin levels in the shoot base and roots are lower compared to controls. The greatest effect (a 39% reduction in the shoot base) is produced at an herbicide concentration of 10 milligrams per liter. Exogenous 10 micromolar ABA greatly stimulates lectin accumulation in the presence of fluridone, but the levels are not as high as those produced by ABA alone. These results indicate that lectin synthesis is under ABA control in both wheat embryos and adult plants.


The translocation of focal adhesion kinase in brain synaptosomes is regulated by phosphorylation and actin assembly.
Focal adhesion kinase (FAK) and the related proline-rich tyrosine kinase 2 (PYK2) are non-receptor protein tyrosine kinases that transduce extracellular signals through the activation of Src family kinases and are highly enriched in neurones. To further elucidate the regulation of FAK and PYK2 in nervous tissue, we investigated their distribution in brain subcellular fractions and analysed their translocation between membrane and cytosolic compartments. We have found that FAK and PYK2 are present in a small membrane-associated pool and a larger cytosolic pool in various neuronal compartments including nerve terminals. In intact nerve terminals, inhibition of Src kinases inhibited the membrane association of FAK, but not of PYK2, whereas tyrosine phosphatase inhibition sharply increased the membrane association of both FAK and PYK2. Disruption of the actin cytoskeleton was followed by a decrease in the membrane-associated pool of FAK, but not of PYK2. For both kinases, a significant correlation was found between autophosphorylation and membrane association. The data indicate that FAK and PYK2 are present in nerve terminals and that the membrane association of FAK is regulated by both phosphorylation and actin assembly, whereas that of PKY2 is primarily dependent on its phosphorylation state.


[The treatment of urinary stress incontinence with the sling and Pereyra's suspension].
Patients with proven stress incontinence were subjected to surgical treatment which combined both sling and Stamey-Pereyra techniques. A 3-4 cm long and 1,5-2 cm wide sling of Bulgarian polyamide mesh was placed under the urethrovesical junction using a small vaginal incision. Two long polyamide sutures were tied to either end of the mesh and were retrieved from the vagina to the abdomen by Pereyra needle inserted through two small suprapubic incisions and guided by the vaginal forefinger. The technique of the procedure is described. A total number of 27 patients were operated on. Two of them has their suspending sutures cut postoperatively because of prolonged urinary retention. The remaining 25 were continent. The technique proposed combines the advantages of the sling procedures (high efficiency especially in cases of severe and recurrent incontinence) and the simplicity of Stamey-Pereyra suspension technique. On the other hand the Bulgarian polyamide mesh showed very good qualities. No infection, rejection or sling erosion were observed.


Sewage sludge treatment in a thermophilic membrane reactor (TMR): factors affecting foam formation.
Foam formation in the excess sludge treatment facilities of biological wastewater treatment plants (WWTPs) may represent a critical issue as it could lead to several operative problems and reduce the overall plant performance. This trouble also affects a novel technology recently proposed for sludge minimization, the thermophilic membrane reactor (TMR), operating with alternate aeration/non-aeration cycles. This technology, which has proven to be extremely resilient and suitable for treating industrial wastewater of different nature, demonstrated a high potential also as a solution for integrating existing WWTPs, aiming at the "zero sludge production." In this work, an experimental study was conducted with a TMR pilot plant (fed daily with thickened sewage sludge) by adjusting the duration of aeration/non-aeration alternate cycles. Extracellular polymeric substance (EPS) concentration (and its soluble and bound fractions) has been monitored along with foaming power indices. The results highlight that foaming can be correlated to the presence of soluble protein fraction of EPS. Moreover, EPS production seems to be reduced by increasing the duration of the non-aeration cycles: optimal operating conditions resulted 2 h of aeration followed by 6 h of non-aeration. These conditions allow to obtain an EPS concentration of 500 mg L-1 with respect to 2300 mg L-1 measured at the beginning of experimental work.


Identification and analysis of multi-protein complexes in placenta.
Placental malfunction induces pregnancy disorders which contribute to life-threatening complications for both the mother and the fetus. Identification and characterization of placental multi-protein complexes is an important step to integratedly understand the protein-protein interaction networks in placenta which determine placental function. In this study, blue native/sodium dodecyl sulfate polyacrylamide gel electrophoresis (BN/SDS-PAGE) and Liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to screen the multi-protein complexes in placenta. 733 unique proteins and 34 known and novel heterooligomeric multi-protein complexes including mitochondrial respiratory chain complexes, integrin complexes, proteasome complexes, histone complex, and heat shock protein complexes were identified. A novel protein complex, which involves clathrin and small conductance calcium-activated potassium (SK) channel protein 2, was identified and validated by antibody based gel shift assay, co-immunoprecipitation and immunofluorescence staining. These results suggest that BN/SDS-PAGE, when integrated with LC-MS/MS, is a very powerful and versatile tool for the investigation of placental protein complexes. This work paves the way for deeper functional characterization of the placental protein complexes associated with pregnancy disorders.


[Assessment of the efficiency of population policy measures].
This article is concerned with the relative effectiveness of various population policy measures. "The data sources [for] analysing efficiency (vital statistics, census data, longitudinal and retrospective analysis of fertility, data of public opinion research) and the possible methods (standardization, trend analysis, experimental samples, number of years spent for contraception by co-habitants, component forecasting approach, the analysis of the reproduction process, regression analysis, simulation models) are reviewed." (summary in ENG, RUS)


Insulin binds to type V collagen with retention of mitogenic activity.
The abilities of eight extracellular matrix proteins, fibronectin, vitronectin, laminin, and collagen types I, II, III, IV, and V to bind insulin were examined by binding studies with insulin conjugated with peroxidase. At a physiological pH and ionic strength, type V collagen bound to insulin most strongly. The other types of collagen, laminin, and vitronectin also bound insulin with affinity lower than that of type V collagen. The insulin-binding site of type V collagen was in a 30-kDa CNBr fragment of the alpha 1 (V) chain. Analysis of the amino acid sequence showed that this 30-kDa fragment was identical to the heparin-binding fragment of type V collagen. The insulin-binding sites of laminin and vitronectin were located in the A chain and in the heparin-binding domain, respectively. Insulin bound to type V collagen stimulated the synthesis of DNA by mouse mammary tumor MTD cells, indicating that bound insulin retained mitogenic activity.


Phase transitions in diluted negative-weight percolation models.
We investigate the geometric properties of loops on two-dimensional lattice graphs, where edge weights are drawn from a distribution that allows for positive and negative weights. We are interested in the appearance of spanning loops of total negative weight. The resulting percolation problem is fundamentally different from conventional percolation, as we have seen in a previous study of this model for the undiluted case. Here, we investigate how the percolation transition is affected by additional dilution. We consider two types of dilution: either a certain fraction of edges exhibits zero weight, or a fraction of edges is even absent. We study these systems numerically using exact combinatorial optimization techniques based on suitable transformations of the graphs and applying matching algorithms. We perform a finite-size scaling analysis to obtain the phase diagram and determine the critical properties of the phase boundary. We find that the first type of dilution does not change the universality class compared to the undiluted case whereas the second type of dilution leads to a change in the universality class.


[High-grade non-Hodgkin lymphoma: diagnosis and therapy].
High grade non-Hodgkin's lymphomas comprise a group of heterogenous disorders of the lymphatic system with aggressive clinical behaviour. Extranodal disease manifestations are common, especially in immunocompromised patients [e.g. AIDS]. High-grade NHL are chemo- and radiosensitive. Anthracyclin-containing chemotherapy regimens have led to a significant improvement in prognosis. The CHOP-regime, consisting of doxorubicin, cyclophosphamide, oncovin and prednisolon, is the standard therapy that should be given outside of clinical trials. Age, bad performance status, elevated serum-LDH, presence of more than one extranodal manifestation and disease stage III or IV have been defined as clinically relevant prognostic factors. Current treatment strategies include dose intensification by interval shortening and dose escalation as well as high-dose chemotherapy, followed by autologous stem cell rescue. Patients receiving these experimental therapies should be treated within multicenter clinical trials.


Effects of atropine and soman on the pharmacokinetics of the oxime HLö 7 dimethanesulfonate in anesthetized guinea-pigs.
The respiratory and circulatory effects and the pharmacokinetics of the bispyridinium dioxime HLö 7 (30 mumol/kg) were investigated in guinea-pigs in combination with atropine (28.8 mumol/kg) or with atropine plus soman (0.44 mumol/kg = 5 x LD50). The pharmacokinetics and pharmacodynamics were studied in separate groups of animals. HLö 7 had only minor effects on circulation and respiration. HLö 7 plus atropine caused a transient, pronounced fall of blood pressure, tachycardia, and respiratory stimulation. The soman-induced bradycardia was completely restored by the antidotes, while respiration improved within a few minutes with the respiratory rate becoming stabilized at 50% of baseline. Plasma half-time (60-70 min), plasma clearance (6-7 ml.kg-1.min-1) and apparent volume of distribution (0.5-0.6 l/kg) of HLö 7 did not differ between the groups, i.e. atropine and soman did not affect the kinetics of the oxime.


Characterization of a monoclonal antibody against desmoplakin 1 for use in dogs.
Skin biopsy specimens from face, axilla, abdomen and thigh, mucocutaneous tissues from anus and vagina, and oral mucosa from six healthy Beagle dogs were examined for desmoplakin (Dsp) immunoreactivity using immunoblotting and immunohistochemical analysis. With immunoblotting using mouse antihuman Dsp 1 monoclonal antibody (DP2.17), a band was detected at 250 kDa in all the extracts as in normal human skin samples, although no band was detected at 210 kDa, suggesting that monoclonal antibody DP2.17 recognizes canine Dsp 1 but not Dsp 2. Moreover, the desmosome regions of all specimens were stained with DP2.17 using immunohistochemical analysis. From these results, DP2.17, developed for the examination of human skin, might be suitable for the investigation of Dsp-related skin disorders in dogs.


Vibrio jasicida sp. nov., a member of the Harveyi clade, isolated from marine animals (packhorse lobster, abalone and Atlantic salmon).
Six isolates of a facultatively anaerobic bacterium were recovered in culture from marine invertebrates and vertebrates, including packhorse lobster (Jasus verreauxi), abalone (Haliotis sp.) and Atlantic salmon (Salmo salar), between 1994 and 2002. The bacteria were Gram-negative, rod-shaped and motile by means of more than one polar flagellum, oxidase-positive, catalase-positive and able to grow in the presence of 0.5-8.0% NaCl (optimum 3.0-6.0%) and at 10-37 °C (optimum 25-30 °C). On the basis of 16S rRNA gene sequence analysis and multilocus sequence analysis (MLSA) using five loci (2443 bp; gyrB, pyrH, ftsZ, mreB and gapA), the closest phylogenetic neighbours of strain TCFB 0772(T) were the type strains of Vibrio communis (99.8 and 94.6 % similarity, respectively), Vibrio owensii (99.8 and 94.1%), Vibrio natriegens (99.4 and 88.8%), Vibrio parahaemolyticus (99.4 and 90.3%), Vibrio rotiferianus (99.2 and 94.4%), Vibrio alginolyticus (99.1 and 89.3%) and Vibrio campbellii (99.1 and 92.3%). DNA-DNA hybridization confirmed that the six isolates constitute a unique taxon that is distinct from other known species of Vibrio. In addition, this taxon can be readily differentiated phenotypically from other Vibrio species. The six isolates therefore represent a novel species, for which the name Vibrio jasicida sp. nov. is proposed; the novel species is represented by the type strain TCFB 0772(T) ( = JCM 16453(T) = LMG 25398(T)) (DNA G+C content 45.9 mol%) and reference strains TCFB 1977 ( = JCM 16454) and TCFB 1000 ( = JCM 16455).


Accuracy of Intraocular Lens Power Calculation Formulas for Highly Myopic Eyes.
Purpose. To evaluate and compare the accuracy of different intraocular lens (IOL) power calculation formulas for eyes with an axial length (AL) greater than 26.00 mm. Methods. This study reviewed 407 eyes of 219 patients with AL longer than 26.0 mm. The refractive prediction errors of IOL power calculation formulas (SRK/T, Haigis, Holladay, Hoffer Q, and Barrett Universal II) using User Group for Laser Interference Biometry (ULIB) constants were evaluated and compared. Results. One hundred seventy-one eyes were enrolled. The Barrett Universal II formula had the lowest mean absolute error (MAE) and SRK/T and Haigis had similar MAE, and the statistical highest MAE were seen with the Holladay and Hoffer Q formulas. The interquartile range of the Barrett Universal II formula was also the lowest among all the formulas. The Barrett Universal II formulas yielded the highest percentage of eyes within ±1.0 D and ±0.5 D of the target refraction in this study (97.24% and 79.56%, resp.). Conclusions. Barrett Universal II formula produced the lowest predictive error and the least variable predictive error compared with the SRK/T, Haigis, Holladay, and Hoffer Q formulas. For high myopic eyes, the Barrett Universal II formula may be a more suitable choice.


Pitfalls and recommendations in cases of laser removal of decorative tattoos with pigmented lesions: case report and review of the literature.
Pigmented lesions in decorative tattoos cause diagnostic difficulties at a clinical and dermoscopic level. In cases of laser removal of tattoos, hidden suspicious nevi may be revealed gradually. We describe the first case of a malignant melanoma that developed on a preexisting nevus within a tattoo during and between the phases of laser removal. The patient refused to undergo excision of the nevus until we made excision conditional for continued laser treatment. The English literature reports 16 cases of malignant melanoma developing in tattoos. Correlation between the placement of tattoos and the development of malignant melanoma remains unclear. Our case emphasizes the diagnostic problems of pigmented lesions within tattoos. For safety reasons, tattoos should never be placed on pigmented lesions; if they are, the tattoos should not undergo laser treatment. We suggest an excision before starting laser tattoo removal. Dermoscopic assessments on a regular basis during the period of tattoo removal are recommended.


Mouse sperm cryopreservation and recovery using the I·Cryo kit.
Thousands of new genetically modified (GM) strains of mice have been created since the advent of transgenesis and knockout technologies. Many of these valuable animals exist only as live animals, with no backup plan in case of emergency. Cryopreservation of embryos can provide this backup, but is costly, can be a lengthy procedure, and generally requires a large number of animals for success. Since the discovery that mouse sperm can be successfully cryopreserved with a basic cryoprotective agent (CPA) consisting of 18% raffinose and 3% skim milk, sperm cryopreservation has become an acceptable and cost-effective procedure for archiving, distributing and recovery of these valuable strains. Here we demonstrate a newly developed I•Cryo kit for mouse sperm cryopreservation. Sperm from five commonly-used strains of inbred mice were frozen using this kit and then recovered. Higher protection ratios of sperm motility (> 60%) and rapid progressive motility (> 45%) compared to the control (basic CPA) were seen for sperm frozen with this kit in 5 inbred mouse strains. Two cell stage embryo development after IVF with the recovered sperm was improved consistently in all 5 mouse strains examined. Over a 1.5 year period, 49 GM mouse lines were archived by sperm cryopreservation with the I•Cryo kit and later recovered by IVF.


[Other malignant tumors of the pleura].
Malignant tumors of the pleura are most often diffuse, nethertheless they are sometimes localized. There is an overlap of the radiologic features of the benign and malignant pleural lesions. The differential diagnosis may be difficult, even on histological sample. Imaging allows the diagnosis of pleural involvement, suggests the malignity, guides percutaneous or thoracoscopic biopsies of the pleura, defines extent of the tumor and follows the course of the disease. We will describe the malignant pleural tumors: pleural metastases, pleural involvement of broncho-pulmonary cancer, of lymphoma and leukaemia. Then the rare pleural tumors will be described: malignant pleural fibroma, sarcoma, histiocytoma and hemangiopericytoma.


Physical exercise and synaptic protection in human and pre-clinical models of multiple sclerosis.
In multiple sclerosis, only immunomodulatory and immunosuppressive drugs are recognized as disease-modifying therapies. However, in recent years, several data from pre-clinical and clinical studies suggested a possible role of physical exercise as disease-modifying therapy in multiple sclerosis. Current evidence is sparse and often conflicting, and the mechanisms underlying the neuroprotective and antinflammatory role of exercise in multiple sclerosis have not been fully elucidated. Data, mainly derived from pre-clinical studies, suggest that exercise could enhance long-term potentiation and thus neuroplasticity, could reduce neuroinflammation and synaptopathy, and dampen astrogliosis and microgliosis. In humans, most trials focused on direct clinical and MRI outcomes, as investigating synaptic, neuroinflammatory, and pathological changes is not straightforward compared to animal models. The present review analyzed current evidence and limitations in research concerning the potential disease-modifying therapy effects of exercise in multiple sclerosis in animal models and human studies.


Relationship of Helicobacter pylori infection to several malignant and non-malignant gastrointestinal diseases.
The importance of the Helicobacter pylori infection was investigated as a risk factor for several gastrointestinal diseases. In this study 203 patients with gastric cancer, 61 with peptic ulcus, 60 with gastritis and 100 asymptomatic control subjects were investigated. Serum samples were examined for IgC antibodies to H. pylori by enzyme linked immunoassay - tissue samples were stained for H. pylori by Wartin-Stary technique and by Giemsa for routine histopathology. H. pylori seropositivity was 58.1% in gastric cancer, 54% in peptic ulcus, 63.3% in gastritis and 27% in asymptomatic control group. There was a 10.1% discordance between the serum and tumor determinants in the seropositive group and 11.3% of discordance in the seronegative group. The frequency of H. pylori seropositivity was lowest in cardia tumors (22.7%) and highest in antral tumors (65.5%, p=0.00002). H. pylori seropositivity was 29% in diffuse type of histology, 35% in mixed type and 79% in the intestinal type (p=0.00000). In the gastric cancer patients the frequent use of salty food (p=0.00001, OR=6.4), excessive salt, pickled food (p=0.0000, OR=24.92) and proteins (p=0.003, OR=0.45) were more significant than asymptomatic volunteers. In gastric cancer patients the frequent use of salty and pickled food were relevantly associated with H. pylori infection (p=0.001). It was concluded that H. pylori infection could play a role in the pathogenesis of non-malignant gastrointestinal diseases which may be the precursor of carcinoma. However, other contributing factors to carcinogenesis must be investigated.


Reckless males, rational females: dynamic trade-off between food and shelter in the marine isopod Idotea balthica.
Habitat choice of herbivores is expected to be a resolution of a trade-off between food and shelter. The resolution of this trade-off may, however, be dynamic within a species because distinct phenotypes may value these factors differently and the value may vary temporally. We studied this hypothesis in the marine herbivore Idotea balthica (Isopoda), by simultaneously manipulating both food and shelter, and investigated whether the resolution of the trade-off differed between sexes, colour morphs and day and night (i.e. high and low predation risk). Isopods chose between exposing and concealing backgrounds in which the quantity or quality of food varied. When choosing between the backgrounds in the absence of food, females preferred the concealment more than males did. However, in a trade-off situation the isopods traded shelter for food, and females more so than males. Thus, males' lower preference for the shelter was not counterbalanced by a stronger preference for food. The microhabitat use also differed between night and day showing adaptation to diurnally fluctuating predation risk. We suggest that microhabitat utilization of females is more strongly tied to variation in risk and resources than that of males, for whom other factors, such as seeking mates, may be more important.


Characterization by image cytometry of duct epithelial proliferative disease of the breast.
To develop a morphometric model of premalignant breast epithelium, we evaluated 120 lesions classified as nonproliferative disease (n = 20), hyperplasia (n = 20), moderate hyperplasia (n = 20), atypical hyperplasia (n = 20), carcinoma in situ (n = 20), and carcinoma (n = 20) in tissue from surgical biopsy or mastectomy. Atypical hyperplasia, a component of duct epithelial proliferative disease, has frequently been described in breasts with carcinoma. Atypical hyperplasia is generally viewed as premalignant or as a marker of increased risk for breast cancer. Measurements of nuclei in breast lesions were obtained with the Leitz TAS Plus on 4-microns sections stained for DNA with the Azure A Feulgen reaction. Nuclei of duct epithelial lesions had morphometric features that displayed changes from nonproliferative disease to carcinoma. The morphometric data from each lesion were compared among the six disease groups. Means of nuclear area, perimeter, maximum and minimum diameter, and large dark and large light intranuclear areas increased with higher degrees of proliferative abnormality. When the six groups of lesions were compared using the means of the first four nuclear features, atypical hyperplasia was significantly different (P less than 0.05) from carcinoma and non-proliferative lesions, but not from hyperplasia, moderate hyperplasia, or carcinoma in situ. These findings suggest that objective morphometric descriptors for characterizing significant proliferative lesions can be established using image cytometry. The progressive increases also suggest that proliferative breast disease is a continuum that includes premalignant lesions.


Rapid isolation of endosomes from BHK cells: identification of DPP IV (CD26) in endosomes.
In plasma membrane glycoproteins the peripheral monosaccharides of the N-glycan side chains are degraded faster than the core oligosaccharides and the protein backbone. This intramolecular heterogeneous turnover is a typical characteristic of membrane glycoproteins and is termed remodeling or reprocessing. The mechanism of the reprocessing has been shown first for dipeptidyl peptidase IV (DPP IV, CD26). However, it is still a question in which subcellular compartment the enzyme machinery for the reprocessing is located. Since lysosomes could be excluded, it has been proposed that the responsible glycosidases are located at the plasma membrane, in endosomes, or in the trans-Golgi network. The present study is concerned with the possible role of endosomes in this process of reprocessing. We transfected nonpolarized BHK cells with rat DPP IV cDNA. By establishing a fast and efficient method to purify endosomes, we could identify for the first time significant amounts of DPP IV in endosomes and we suggest therefore that endosomes are closely related with the regulation of reprocessing of plasma membrane glycoproteins.


Silencing of methyltransferase-like 3 inhibits oesophageal squamous cell carcinoma.
Methyltransferase-like 3 (METTL3) is a methyltransferase responsible for N6-methyladenosine mRNA modifications, which has been demonstrated to serve oncogenic roles in various types of cancer; however, the exact function of METTL3 in oesophageal squamous cell carcinoma (ESCC) has not been determined. The present study aimed to explore the regulatory role of METTL3 in ESCC. In the present study, reverse transcription-quantitative PCR and western blotting were used to examine mRNA and protein expression, CCK-8 assays and flow cytometry were used to determine cellular viability and apoptosis, and wound healing and Transwell assays were conducted to study cellular migration and invasion. The expression levels of METTL3 were significantly higher in ESCC tissues and cell lines compared with adjacent non-tumour tissues and the normal oesophageal epithelial cell line HET-1A, respectively. Increased METTL3 expression was associated with an advanced clinical stage of ESCC and poorer prognosis. Furthermore, the genetic knockdown of METTL3 using small interfering RNA significantly suppressed ESCC growth, invasion and migration in vitro, and induced cellular apoptosis, in addition to reducing the phosphorylation levels of PI3K and AKT. In conclusion, the present study demonstrated that the upregulation of METTL3 promoted ESCC progression, and that inhibition of METTL3 significantly suppressed the malignant phenotypes of ESCC cells, at least in part, by downregulating PI3K/AKT signalling activity. Thus, it is suggested that METTL3 may be a promising therapeutic target for ESCC.


Diagnostic relevance of suppressed basal concentrations of TSH compared with the negative TRH test in detection and exclusion of hyperthyroidism.
To evaluate the sensitivity of basal TSH concentrations as determined by an "ultrasensitive" IRMA-assay (RIA-gnost h-TSH-monoclonal, Behring) versus a "negative" TRH test (defined as an increment of TSH less than or equal to 0.2 mU/l 20 min after administration of 400 micrograms TRH iv) in the diagnosis of hyperthyroidism we examined 193 consecutive patients from our thyroid outpatient clinic: 34 patients displayed hyperthyroidism (total T4: 184.4 +/- 26.0 mumol/l, effective thyroxine index: 1.25 +/- 0.08), whereas 12 had isolated T3-hyperthyroidism (total T3: 3.47 +/- 0.48 nmol/l). Employing the producer's definition of subnormal ("suppressed") bTSH concentrations (less than or equal to 0.1 mU/l), only 19 (41.3%) hyperthyroid patients would have been detected; on the other hand, one euthyroid patient would have been recognized false positively as hyperthyroid. Using the TRH test as criterion led to the correct diagnosis in 42 (sensitivity: 91.3%) hyperthyroid patients, whereas two had low bTSH concentrations (less than or equal to 0.5 mU/l), but a normal TSH response to TRH (greater than 2.0 mU/l). Raising the threshold concentration to 0.2 and, subsequently, to 0.4 mU TSH/l increased the number of correct results to 38 (sensitivity: 82.6%) and 43 (93.5%), respectively. This was associated with a concomitant decrease in specificity in the diagnosis of hyperthyroidism from 93.7 (0.1 mU/l) to 27.9% (0.4 mU/l). In conclusion, despite ultrasensitive methods for estimation of low TSH concentrations, the TRH test remains an irreplaceable tool for the correct diagnosis of hyperthyroidism.


Microencapsulated sodium fusidate to impart antibacterial properties into cotton dressings for traumatic wounds.
This research aimed to investigate the use of microencapsulated sodium fusidate to impart antibacterial properties into conventional cotton gauze used to treat traumatic wounds. The microcapsules were prepared with sodium fusidate dissolved in ethanol as the active substance (core) and calcium alginate as the wall material (shell) using an electrospraying method. The microcapsules were then loaded into a 1% solution of chitosan, and conventional sterile cotton gauze fabric was impregnated in this suspension and dried at room temperature. The microcapsules obtained were examined with Fourier transform infrared spectroscopy to confirm the constituents, and by scanning electron microscopy to examine the size and distribution of the microcapsules. The results showed that microcapsules with sodium fusidate as the core material were produced in a size range of 50-200µm. The antibacterial properties of these gauze samples were tested for resistance to both Gram-positive and Gram-negative bacteria. The samples showed antibacterial properties against Gram-positive bacteria (Staphylococcus aureus) only, with an average inhibition zone of 31.67±2.89mm in diameter. The antibacterial effect of the treated gauze samples was tested after washing to investigate the strength of the binding agent, to find out whether the samples maintained antibacterial properties after two hand-rubbed washes. Its absorption capacity, rate of absorbency and air permeability values were 555±4.51%, 1.25±0.04 seconds and 408±2.65cm3/cm2/seconds, respectively. The research findings have established the possibility of imparting antibacterial properties into sterile cotton gauze using microencapsulation of sodium fusidate.


Defining a reference set to support methodological research in drug safety.
Methodological research to evaluate the performance of methods requires a benchmark to serve as a referent comparison. In drug safety, the performance of analyses of spontaneous adverse event reporting databases and observational healthcare data, such as administrative claims and electronic health records, has been limited by the lack of such standards. To establish a reference set of test cases that contain both positive and negative controls, which can serve the basis for methodological research in evaluating methods performance in identifying drug safety issues. Systematic literature review and natural language processing of structured product labeling was performed to identify evidence to support the classification of drugs as either positive controls or negative controls for four outcomes: acute liver injury, acute kidney injury, acute myocardial infarction, and upper gastrointestinal bleeding. Three-hundred and ninety-nine test cases comprised of 165 positive controls and 234 negative controls were identified across the four outcomes. The majority of positive controls for acute kidney injury and upper gastrointestinal bleeding were supported by randomized clinical trial evidence, while the majority of positive controls for acute liver injury and acute myocardial infarction were only supported based on published case reports. Literature estimates for the positive controls shows substantial variability that limits the ability to establish a reference set with known effect sizes. A reference set of test cases can be established to facilitate methodological research in drug safety. Creating a sufficient sample of drug-outcome pairs with binary classification of having no effect (negative controls) or having an increased effect (positive controls) is possible and can enable estimation of predictive accuracy through discrimination. Since the magnitude of the positive effects cannot be reliably obtained and the quality of evidence may vary across outcomes, assumptions are required to use the test cases in real data for purposes of measuring bias, mean squared error, or coverage probability.


Lysyl oxidase-like 1 deficiency alters ultrastructural and biomechanical properties of the peripapillary sclera in mice.
Lysyl oxidase-like 1 encoded by the LOXL1 gene is a member of the lysyl oxidase family of enzymes that are important in the maintenance of extracellular matrix (ECM)-rich tissue. LOXL1 is important for proper elastic fiber formation and mice lacking LOXL1 (Loxl1-/- ) exhibit systemic elastic fiber disorders, such as pelvic organ prolapse, a phenotype associated with exfoliation syndrome (XFS) in humans. Patients with XFS have a significant risk of developing exfoliation glaucoma (XFG), a severe form of glaucoma, which is a neurodegenerative condition leading to irreversible blindness if not detected and treated in a timely fashion. Although Loxl1-/- mice have been used extensively to investigate mechanisms of pelvic organ prolapse, studies of eyes in those mice are limited and some showed inconsistent ocular phenotypes. In this study we demonstrate that Loxl1-/- mice have significant anterior segment biometric abnormalities which recapitulate some human XFS features. We then focused on the peripapillary sclera (PPS), a critical structure for maintaining optic nerve health. We discovered quantitative and qualitive changes in ultrastructure of PPS, such as reduced elastic fibers, enlarged collagen fibrils, and transformed collagen lamella organization detected by transmission electron microscopy (TEM). Importantly, these changes corelate with altered tissue biomechanics detected by Atomic Force Microscopy (AFM) of PPS in mice. Together, our results support a crucial role for LOXL1 in ocular tissue structure and biomechanics, and Loxl1-/- mice could be a valuable resource for understanding the role of scleral tissue biomechanics in ocular disease.


Dengue virus infection in renal allograft recipients: a case series during 2010 outbreak.
Dengue virus infection is an emerging global threat caused by Arbovirus, a virus from Flaviridiae family, which is transmitted by mosquitoes, Aedes aegypti and Aedes albopictus. Renal transplant recipients who live in the endemic zones of dengue infection or who travel to an endemic zone could be at risk of this infection. Despite multiple epidemics and a high case fatality rate in the Southeast Asian region, only a few cases of dengue infection in renal transplant recipients have been reported. Here, we report a case series of 8 dengue viral infection in renal transplant recipients. Of the 8 patients, 3 developed dengue hemorrhagic shock syndrome and died.