Progressive multifocal leukoencephalopathy in a patient with acquired immune deficiency syndrome.
Progressive multifocal leukoencephalopathy (PML) occurred in a heterosexual Haitian man with acquired immune deficiency syndrome (AIDS). The patient initially had focal neurologic signs and nonenhancing lesions on a computed tomographic scan. Although PML is rare, it should be included in the differential diagnosis of opportunistic infections associated with AIDS. Brain biopsy is suggested in patients suspected of having PML who might benefit from antiviral therapy.


The simple bunion: anatomy at the metatarsophalangeal joint of the great toe.
The pathomechanics for the development of the hallux valgus deformity takes place at the first metatarsophalangeal joint--the sesamoid complex. The sesamoid complex consists of seven muscles, eight ligaments, and two sesamoid bones. When the first metatarsal escapes the complex and drifts medially, the sesamoids remain twisted in situ, several of the ligaments "fail," and others contract. The authors propose reduction of the metatarsus primus varus by first metatarsal osteotomy and appropriate ligament releases and plications to restore alignment. A detailed understanding of the pathomechanics is essential for proper interpretation of the problems and anticipated lasting surgeries.


Business English Translation Model Based on BP Neural Network Optimized by Genetic Algorithm.
In recent years, most of the communication places are using business English manual simultaneous interpretation or electronic equipment translation. In the context of diverse cultures, the way English is used and its grammar vary from country to country. In the face of this situation, how to optimize business English translation technology and improve the accuracy of business communication content is one of the research contents of scholars all over the world. This paper first introduces the purpose of business English translation and the gap between business English translation and general English translation. Secondly, a genetic algorithm is used to optimize the structure of the BP neural network, and the combination of the two improves the ability of translation search. This paper compares the influence of the traditional BP algorithm and the BP algorithm optimized by genetic algorithm on the construction of a business English translation model. The results show that BP neural network optimized by the genetic algorithm can improve the speed of business English text translation, reduce the impact of semantic errors on the accuracy of the translation model, and improve the efficiency of translation.


Radicals and molecular products from the gas-phase pyrolysis of lignin model compounds. Cinnamyl alcohol.
The experimental results on detection and identification of intermediate radicals and molecular products from gas-phase pyrolysis of cinnamyl alcohol (CnA), the simplest non-phenolic lignin model compound, over the temperature range of 400-800 °C are reported. The low temperature matrix isolation - electron paramagnetic resonance (LTMI-EPR) experiments along with the theoretical calculations, provided evidences on the generation of the intermediate carbon and oxygen centered as well as oxygen-linked, conjugated radicals. A mechanistic analysis is performed based on density functional theory to explain formation of the major products from CnA pyrolysis; cinnamaldehyde, indene, styrene, benzaldehyde, 1-propynyl benzene, and 2-propenyl benzene. The evaluated bond dissociation patterns and unimolecular decomposition pathways involve dehydrogenation, dehydration, 1,3-sigmatropic H-migration, 1,2-hydrogen shift, C-O and C-C bond cleavage processes.


The History of Surgical Education in the United States: Past, Present, and Future.
In just over 100 years, surgical education in the United States has evolved from a disorganized practice to a refined system esteemed worldwide as one of the premier models for the training of physicians and surgeons. But in the changing environment of health care, new challenges have arisen that could warrant a reform. To design our future, we must understand our past. The present work is not intended to be a comprehensive account of the history of American surgery. Instead, it tells the abridged history of surgical education in our country: the evolution from apprenticeships to residencies; the birth of hospital-based teaching; the impact of key historical events on training; the marks left by some preeminent characters; the conception of regulatory entities that steer our education; and, finally, how our process of training surgeons might need to be refined for the continued progress of our profession. Told in chronological order in a manner that will be memorable to readers, this story weaves together the key events that explain how our current surgical training models came to be. We conclude with a timely invitation to draw from these past lessons to redesign the future of graduate medical education, making a case for the transition to time-variable, competency-based medical education for surgical residency programs in America.


Induction of idiotype-bearing, nuclease-specific helper T cells by in vivo treatment with anti-idiotype.
Treatment of BALB/c mice with purified pig anti-(BALB/c anti-nuclease) anti-idiotypic antibodies has been found to induce the appearance of idiotype-bearing immunoglobulins (Id') in the serum of these mice in the absence of detectable antigen binding activity. This phenomenon appeared to require T cells in the hosts because no Id' was detected in the serum of nude mice similarly treated. Furthermore, the spleens of BALB/c mice treated with anti-idiotype were found to contain helper T cells capable of providing help in an in vitro plaque-forming cell response to trinitrophenyl-nuclease equivalent to that provided by helper T cells from the spleens of nuclease-primed animals. Helper T cells from both anti-idiotype-treated and nuclease-treated animals were found to be antigen-specific and to be similarly susceptible to elimination by treatment with anti-idiotype plus complement. Therefore, treatment with both antigen and anti-idiotype appeared to prime similar populations of antigen-specific helper T cells, while having different effects on the induction of antibody. These findings are consistent with the network theory of receptor interactions in the immune response, and may provide a means for studying individual cell populations involved in such interactions.


Gold-Catalyzed Domino Synthesis of Functionalized Benzofurans and Tetracyclic Isochromans via Formal Carboalkoxylation.
A domino synthesis of benzofurans with the modification of side chains from α-alkoxyalkyl o-alkynylaryl ethers (n = 0) and electron-rich arenes has been developed. In the present domino reaction, which would proceed via the α-alkoxyalkylation of arenes with an intermediate in the migratory cycloisomerization of o-alkynylaryl ethers followed by the nucleophilic addition of benzofurans to benzyl ethers, a cationic Au(III) catalyst activates the C-C π bond and the C-O σ bond. The present method could be extended to Au(I)-catalyzed domino synthesis of tetracyclic isochromans from α-alkoxyalkyl (o-alkynylaryl)methyl ethers (n = 1) and aryl methoxymethyl ethers.


Comparison of radiofrequency ablation, cryoablation, and nephrectomy in treating implanted VX-2 carcinoma in rabbit kidneys.
To compare the efficacy of radiofrequency (RF) ablation, cryoablation, and radical nephrectomy in the treatment of implanted VX-2 carcinoma in rabbit kidneys. Sixty-eight New Zealand White rabbits were implanted with 1-mm3 segments of VX-2 carcinoma in the left kidney. Seven days after implantation, the tumors were treated with one of the following: (1) RF ablation using a 12-gauge electrode (RITA Medical Systems, Mountain View, CA) at 90 degrees C for 8 minutes with a 5-mm tumor margin target temperature of 60 degrees C (N = 20); (2) cryoablation using a 15 minute double-freeze technique with 2.4-mm cryoprobes and the Cryocare system (Endocare Inc., Irvine, CA) with a 5-mm tumor margin target temperature of -20 degrees C (N = 20); (3) open radical nephrectomy (N = 20); or (4) no treatment (controls; N = 8). Rabbits were allowed to survive for a total of 22 days and sacrificed; and the kidneys, lungs, liver, spleen, urinary bladder, and ureter were removed and examined grossly and histologically for tumor. Findings in animals sacrificed at 15 days post-treatment showed significant differences between all treatment groups and untreated controls (P < 0.002) Using a 3 x 2 chi-square comparison, no differences in disease-free survival were observed between the RF ablation group, the cryoablation group, and the open nephrectomy group (P = 0.72) Radiofrequency ablation, cryoablation, and radical nephrectomy were all efficacious in the treatment of implanted VX-2 renal tumors compared with untreated controls (P = 0.002). No statistically significant difference was found between any of the three treatments.


Antioxidant and hepatoprotective activity of Piper retrofractum against Paracetamol-induced hepatotoxicity in Sprague-Dawley rat.
The ethanol extracts of Piper retrofractum were investigated for antioxidant and hepatoprotective activity. Hepatoprotective activity against paracetamol-induced acute hepatotoxicity was estimated in Sprague-Dawley rat. In DPPH free radical assay the root and stem extracts showed IC50 values at 133 and 91 µg/mL, respectively, while ascorbic acid at 14 µg/mL. Extracts also exhibited hydroxyl radical scavenging activity and reducing power. HPLC-DAD analysis indicated the presence of some polyphenolic compounds. Treatment of extracts significantly reduced the elevated serum levels of GPT (P < 0.01), GOT (P < 0.01) and bilirubin (P < 0.001). Both extracts restored the reduced level of total proteins and albumin. A significant increase in HDL-c but decrease in LDL-c level was observed compared to induced control. In histopathological study of liver sections, both extracts showed minimal to mild multifocal and diffuse granular degeneration and mild to moderate lobular disarray compared to control group. Results suggest that both extracts can prevent paracetamol induced hepatotoxicity.


Overweight in young males reduce fertility in rabbit model.
Semen quality has certainly declined over the past few decades, possibly owing to modern lifestyle factors. In this sense, the role of overweight and obesity in the development of subfertility in males has generated a considerable amount of interest in recent years. However, there is no consensus on whether overweight or obesity impaired sperm quality. Thus, based on the ongoing debate about risk factors for subfertility associated with overweight and obesity in men, this study was designed to investigate the effect of overweight on sperm quality parameters and fertility success in randomized controlled trial in a rabbit model. Fourteen male rabbits were randomly assigned to a control group in which nutritional requirements were satisfied or a group fed to satiety from 12 to 32 weeks of age. At 24 weeks of age, semen samples were analysed weekly by conventional semen analysis for 8 weeks. In addition, during the trial female rabbits were artificially inseminated by each male to assess the fertility success and the number of offspring. Young males fed to satiety were associated with a significant increase in body weight (13.6% overweight) and perirenal fat thickness (5%). Male overweight presented a significant decrease in sperm concentration. There were no differences in the remaining sperm parameters. However, male overweight showed a clear and significant decrease in fertility success (control group, 64±8.9% versus fed to satiety group, 35±9.2%), but not in the number of offspring. Taken together, our findings provide new evidence on the loss of fertility induced by overweight in males.


RON and cisplatin resistance in ovarian cancer cell lines.
RON (recepteur d'origine nantais) tyrosine kinase receptor has revealed its tumorigenic potential in recent studies. RON was reported to be overexpressed in 55% of primary ovarian carcinoma samples and furthermore its activation increases cell motility and invasiveness. In this study, we investigated the correlation between RON expression and chemoresistance in ovarian cancer cells. In A2780 cells, a model featured by high chemosensitivity to cisplatin, stable overexpression of RON was able to reduce sensitivity to this agent, while incubation with a blocking anti-RON antibody (ID1) increased the cisplatin-induced growth inhibition effect. Moreover, we observed an increased RON expression both at the mRNA and protein level in A2780 cells made resistant to doxorubicin and paclitaxel (A2780ADR and TC 1, respectively), two cell lines exhibiting a collateral resistance to cisplatin. OVCAR-3 cells, showing high levels of RON expression, also displayed inherent cisplatin resistance. The morphology observed in these resistant cells is consistent with a scattering phenotype and a RON-activated state. RON expression levels were monitored upon hypoxia. A 2.5-fold increase of RON expression was noticed in response to hypoxia in OVCAR-3 cells, in parallel with a decrease of E-cadherin mRNA. Altogether these results suggest an involvement of RON in the acquisition of cisplatin resistance and highlight the importance of this factor as a promising target for combination with cisplatin-based chemotherapy in ovarian cancer.


Myeloid Blast Crisis of Chronic Myeloid Leukemia Followed by Lineage Switch to B-Lymphoblastic Leukemia: A Case Report.
Lineage switch is very rare in blastic crisis of chronic myeloid leukemia (CML-BC). Here, we report a case of CML-BC in which the blast lineage switched from myeloid to B-lymphoid. A 35-year-old male was initially admitted to our hospital because of abdominal distention for over a year and dizziness for one week. Prior to presentation at our hospital, he visited a local hospital because of abdominal distention where his white blood cell count and bone marrow (BM) smear indicated CML. Results from peripheral blood (PB) counts, bone marrow analysis, immunophenotyping by flow cytometry, and the detection of the Philadelphia chromosome were consistent with a diagnosis of myeloid blast crisis from CML. The patient received chemotherapy with imatinib for induction, which diminished the number of blasts. However, after three months, the blasts were increased in the PB and BM. The BM study and immunophenotyping by flow cytometry revealed B-lymphoblastic leukemia. In accordance with his first admission, a chromosome study revealed a karyotype of 46, XY, t(9; 22)(q34; q11) in all 20 cells analyzed, and B-lymphoblastic transformation from CML was diagnosed. Despite three months of treatment with DVCP (daunorubicin, vincristine, cyclophosphamide and prednisone) chemotherapy in combination with dasatinib, the patient did not achieve complete remission. The patient decided to stop treatment and was discharged from the hospital for financial reasons. This case implicates the Philadelphia chromosome with p210 BCR-ABL1 fusion proteins as a key molecule in CML-BC. Further research is needed to assess the frequency, treatment, and prognosis of CML-BC patients with lineage switch.


Health outcomes of vitamin D. Part II. Role in prevention of diseases.
Apart from the classic role of vitamin D, its hormonal active form, calcitriol is also characterized by pleiotropic effects on various organs and tissues. For the last several years, many researchers have shown an association between deficiency of vitamin D and the risk of type 2 diabetes mellitus (T2DM). Recent investigations suggested the need of vitamin D supplementation in T2DM prevention. It was shown that vitamin D deficiency decreases insulin secretion. It was also observed that proper vitamin D supplementation may improve the ability of the cells of the islets of Langerhans to synthesize many proteins de novo and to convert proinsulin to insulin. Apart from regulating bone metabolism and also calcium and phosphate homeostasis, 1,25(OH)2D3 exerts antiproliferative and pro-differentiating effects on a wide variety of cell types. It also induces apoptosis of cancer cells and slows their proliferation. In a number of major studies the relationship between low vitamin D levels and increased risk of various cancers was observed. It concerns colorectal, lung, prostate, breast and ovarian cancer. It was observed that in patients with low serum vitamin D concentrations such disorders as ischemic heart disease, heart attack, stroke, cardiac arrhythmia, and hypertension were more frequent and mortality was significantly higher. These results led the researchers to consider vitamin D deficiency as a potential risk factor for cardiovascular diseases. The possible mechanism in the pathogenesis of cardiovascular diseases that may be related to low levels of vitamin D, is its adverse effect on the renin-angiotensin-aldosterone system (RAAS). Calcitriol is also an important determinant of muscle cell proliferation and differentiation, as well as inhibition of apoptosis. Vitamin D is synthesized in the skin. However, there are only a few food products that are rich in vitamin D3, e.g.: fish oils, fish and fortified-products, such as dairy products and margarines. Individuals who are vulnerable to vitamin D deficiency should be supplemented. vitamin D, diabetes mellitus, deficiency of vitamin D, cancer, cardiovascular diseases, muscle physiology.


Association of an Overhydrated State With the Liver Fibrosis and Prognosis of Cirrhotic Patients.
A bioimpedance analysis (BIA) can indicate an overhydrated state as the extracellular water/total body water (ECW/TBW) value. This study aimed to assess the clinical significance of this value in patients with chronic liver diseases (CLDs). A total of 552 CLD patients who received a liver biopsy and underwent anthropometric assessment and BIA-based body composition analysis were enrolled. The association of the ECW/TBW value with the liver fibrosis and nutritional status was assessed. The relationship between the ECW/TBW value and the prognosis of cirrhotic patients (N=209) was also evaluated. The ECW/TBW value increased as liver fibrosis progressed and was also related to decreased muscle mass/sarcopenia. The presence of overhydration was associated with a poor prognosis of cirrhotic patients. An increased ECW/TBW value was associated with progressive liver fibrosis and malnutrition and related to the prognosis of cirrhotic patients.


[Male circumcision: a banality?].
It is estimated that circumcision concerns approximately 30% of the world male population. Most operations are done for religious or traditional reasons. In clinics, the operation is not without any risk, it has advantage as well as disadvantage. 0.6 to 2% of patients will present complications. However, the risk of contracting a sexually transmitted disease or a Human Immunodeficiency Virus infection is found considerably reduced by circumcised men. Female partners are less risky to develop HIV, Human Papilloma Virus or Chlamydia infection. Circumcision reduced the incidence of penile carcinoma. These data raise the question of the indication of prophylactic circumcision.


The treatment of severe anxiety with chronically implanted intracerebral electrodes.
A procedure is described which allowed discrete lesions to be placed in the frontal lobes and paracingulate white matter through chronically implanted intracerebral electrodes for the treatment of a severe and intractable anxiety state. This procedure enabled progressive treatment on an outpatient basis over a period of weeks so that the amount of treatment was geared carefully to the patient's response. This psychosurgical technique minimizes the risks of untoward side effects. The case history is presented of the first patient treated with chronically implanted intracerebral electrodes at the Neuropsychiatric Institute, Sydney.


Specific engagement of the CD94/NKG2-A killer inhibitory receptor by the HLA-E class Ib molecule induces SHP-1 phosphatase recruitment to tyrosine-phosphorylated NKG2-A: evidence for receptor function in heterologous transfectants.
It has been recently demonstrated that the CD94/NKG2-A killer inhibitory receptor (KIR) specifically recognizes the HLA-E class Ib molecule. Moreover, the apparent CD94-mediated specific recognition of different HLA class Ia allotypes, transfected into the HLA-defective cell line 721.221, indeed depends on their selective ability to concomitantly stabilize the surface expression of endogenous HLA-E molecules, which confer protection against CD94/NKG2-A+ effector cells. In the present study, we show that a selective engagement of the CD94/NKG2-A inhibitory receptor with a specific monoclonal antibody (mAb) (Z199) was sufficient to induce tyrosine phosphorylation of the NKG2-A subunit and SHP-1 recruitment. These early biochemical events, commonly related to negative signaling pathways, were also detected upon the specific interaction of NK cells with an HLA-E+ 721.221 transfectant (.221-AEH), and were prevented by pre-incubation of .221-AEH with an anti-HLA class I mAb. Furthermore, mAb cross-linking of the CD94/NKG2-A receptor, segregated from other NK-associated molecules by transfection into a rat basophilic leukemia cell line (RBL-2H3), promoted tyrosine phosphorylation of NKG2-A and co-precipitation of SHP-1, together with an inhibition of secretory events triggered via Fc epsilonRI. Remarkably, interaction of CD94/NKG2-A+ RBL cells with the HLA-E+ .221-AEH transfectant specifically induced a detectable association of SHP-1 with NKG2-A, constituting a more formal evidence for the receptor-HLA class I interaction.


[Relationship between overweight, obesity and malocclusion in a sample of children].
Obesity and malocclusion are two very common disease worldwide affecting both adults and children, posing a threat to the physical and mental health. Our observational study assesses the impact of being obese and overweight in the development of malocclusion among childhood subjects (3-12 year old).


Reappraisal of histogenesis in the bursal lymphoid follicle of the chicken.
The development of the bursal follicle and the appearance of the follicle-associated epithelial (FAE) cell and the reticuloepithelial (REp) cell were studied. The stages of development of the bursal follicle were observed by light and electron microscopy; an anticytokeratin monoclonal antibody was also used. At the beginning of follicle development, a mesenchymal cell cluster is observed in the tunica propria; the cluster becomes wedged in a niche of the surface epithelium, and gradually it is completely surrounded by the epithelium itself, which closes under the clump of mesenchymal cells. The epithelial cells lying upon the mesenchymal clump become necrotic, and a number of mesenchymal cells bulge out, forming the FAE cells. The epithelial cells that have closed under the mesenchymal nodule become stratified and form the REp cells; they become star-shaped because the medullary-lymphoid cells grow between them. Finally, the cortex is formed, possibly as a result of the migration of medullary cells before they peripheralize. It is concluded that FAE cells are not specialized epithelial cells, as they do not react to an anticytokeratin monoclonal antibody; on the contrary, they are formed by mesenchymal stemcells that bulge into the lumen and change their character after moving into the epithelium. The REp cells appear in the follicular primordium shortly after the bursal follicle begins to develop; the pronounced reactivity of the REp cells to an anticytokeratin monoclonal antibody supports the hypothesis of their epithelial origin.


[Ruptured Left Ventricular Pseudoaneurysm Representing Cardiac Tamponade;Report of a Case].
A 64-year-old woman was followed by the other hospital because of left ventricular false aneurysm after percutaneous coronary intervention for acute myocardial infarction of which culprit lesion was left circumflex artery. She admitted to the hospital suffering from nausea and dyspnea. Echocardiography showed huge pericardial effusion and cardiac tamponade due to ruptured left ventricular false aneurysm and transferred to our hospital under pericardial drainage. She presented with symptoms such as shock, so we performed emergent left ventricular reconstruction. She uneventfully left our hospital on 18 post-operative day.


Congenital left ventricular diverticulum diagnosed by 64-detector CT imaging.
A congenital left ventricular (LV) diverticulum or aneurysm is a rare cardiac malformation. It is a developmental anomaly that occurs during embryogenesis. Most congenital LV aneurysms and diverticula are asymptomatic or may cause systemic embolization, heart failure, valvular regurgitation, ventricular wall rupture, ventricular tachycardia or sudden cardiac death. Diagnosis is established by imaging studies such as echocardiography, magnetic resonance imaging or left ventriculography, visualizing the structural changes. We report the case of a 28-year-old male referred for the evaluation of atypical chest pain who was found to have an LV diverticulum affecting the inferoposterior wall of his LV.


Persistent alteration of pituitary-adrenal function in the rat by prepuberal corticosterone treatment.
The possible existence of periods of sensitivity to elevated levels of corticosterone in the postnatal development of the pituitary-adrenal axis was investigated. Long-Evans rats were implanted ip with corticosterone (25 mg/100 g body wt) mixed with an equal amount of cholesterol on postnatal days 3, 6, 12 or 18. Eight blood samples were obtained from each animal at various ages by cardiac puncture. Steroid treatment during the first postnatal week resulted in lowered adult basal levels of plasma corticosterone without affecting the diurnal rhythm. Males treated on day 3 had lowered peak (PM) and trough (AM) levels of the steroid as adults, while males treated on day 6 showed only a lowered PM level. A similar reduction in basal levels of plasma corticosterone was observed in immature females, but did not persist into adulthood. Steroid treatment at day 12 had no effect on basal or diurnal pituitary-adrenal activity. Treatment at day 18 reduced the amplitude of the adult diurnal rhythm in both sexes and delayed the onset of the rhythm in males. No differences in stress responsiveness or adrenal weights were detected. Thus, two distinct periods exist in the postnatal development of the rat during which basal and diurnal pituitary-adrenal activity can be differentially and persistently altered by treatment with the naturally occurring glucocorticoid.


Task concentration training versus applied relaxation, in combination with cognitive therapy, for social phobia patients with fear of blushing, trembling, and sweating.
Social phobia patients with fear of blushing, trembling, sweating and/or freezing as main complaint (N = 65) were randomly assigned to either task concentration training (TCT) or applied relaxation (AR) both followed by cognitive therapy (CT). Measurements took place before and after wait-list, after TCT or AR (within-test), after CT (post-test), at 3-months and at 1-year follow-up. Effects were assessed on fear of showing bodily symptoms (the central outcome variable), social phobia, other psychopathology, social skills, self-consciousness, self-focused attention, and dysfunctional beliefs. No changes occurred during wait-list. Both treatments were highly effective. TCT was superior to AR in reducing fear of bodily symptoms and dysfunctional beliefs at within-test. This difference disappeared after CT, at post-test and at 3-months follow-up. However, at 1-year follow-up the combination TCT-CT was superior to AR-CT in reducing fear of bodily symptoms, and effect sizes for TCT-CT reached 3. Furthermore, at all assessment moments TCT or the combination TCT-CT was superior to AR-CT in reducing self-consciousness and self-focused attention. The superior long-term effect of TCT on fear of showing bodily symptoms is explained by lasting changes in attentional focus.


Differential regulation of telomerase in endothelial cells by fibroblast growth factor-2 and vascular endothelial growth factor-a: association with replicative life span.
In cultured human umbilical vein endothelial cells (HUVECs), fibroblast growth factor-2 (FGF-2), but not vascular endothelial growth factor-A (VEGF-A), upregulates telomerase activity. Here, we examined the functional significance of this differential regulation on the replicative life span of HUVECs. HUVECs were serially passaged until senescence under four different conditions: (1) EGM-2, a medium containing both VEGF-A and FGF-2; (2) basal medium (BM), consisting of EGM-2 devoid of FGF-2 and VEGF-A; (3) BM supplemented with FGF-2; and (4) BM supplemented with VEGF-A. Cells cultured in BM demonstrated decreased growth rate and ceased to proliferate at approximately 15 population doublings (PDs), whereas those cultured with VEGF-A alone initially proliferated vigorously but arrested growth abruptly at a PD level comparable with cultures grown in BM. In contrast, cells maintained in EGM-2 or in BM/FGF-2 attained a normal replicative life span (approximately 40 PDs). These differences in replicative behavior were reflected by the early appearance of a senescent phenotype in cultures grown in BM or BM/VEGF-A. HUVECs grown in the presence of VEGF-A alone have a decreased life span compared with cultures maintained with FGF-2. This suggests that the upregulation of telomerase activity by FGF-2, an effect not achieved with VEGF-A, plays a functional role in preventing the early onset of senescence.


Pyloric Gland Adenoma of Gallbladder: A Review of Diagnosis and Management.
Neoplastic polypoid mucosal lesions of the gallbladder are increasingly being reported in cholecystectomy specimens. However, due to the absence of unified terminology and reporting criteria, the body of scientific evidence on their classification, prognosis, and management is scarce and sometimes controversial. While they have different histomorphologic features (gastric foveolar, gastric pyloric gland, biliary, and intestinal), a significant immunohistochemical overlap exists which highlights their mixed cell lineage with a dominant cell type in each, establishing the subcategory. Because of many shared attributes, intracholecystic papillary-tubular neoplasm (ICPN) has been introduced as an umbrella terminology. ICPNs of the pyloric subtype are lesions larger than 1 cm, as most of the smaller ones are clinically insignificant and represent polypoid hyperplasia rather than a true neoplasm. In this review, we will focus on the pyloric gland adenomas as the most frequent histologic subtype of ICPNs.


Induction of immunoenhancing factors for murine splenocyte cultures by Salmonella typhimurium ribosome and ribonucleic acid extracts.
Ribosomal and ribonucleic acid (RNA)-rich preparations derived from Salmonella typhimurium were examined for their ability to enhance the primary in vitro antibody response of normal mouse spleen cell cultures to sheep erythrocytes. Both of these fractions were consistently more active in elevating the antibody response of normal mouse splenocytes from lipopolysaccharide (LPS) responder mice than was LPS. Furthermore, injection of mice with either the ribosomal or RNA-rich fraction induced antibody response helper factor activity in 2-h post-treatment serum similar to that induced by LPS. Endotoxin low-responding C3H/HeJ mice were stimulated to release helper factors by ribosomes and the RNA extracts but not by LPS. Treatment of the ribosomes and RNA fractions with ribonuclease destroyed their ability to stimulate the production of the helper factor in serum of treated mice. Therefore, it appears likely that ribosomes and RNA-rich fractions stimulated an intermediate helper factor due to the presence of RNA and not LPS.


2D layered transport properties from topological insulator Bi2Se3 single crystals and micro flakes.
Low-field magnetotransport measurements of topological insulators such as Bi2Se3 are important for revealing the nature of topological surface states by quantum corrections to the conductivity, such as weak-antilocalization. Recently, a rich variety of high-field magnetotransport properties in the regime of high electron densities (∼10(19) cm(-3)) were reported, which can be related to additional two-dimensional layered conductivity, hampering the identification of the topological surface states. Here, we report that quantum corrections to the electronic conduction are dominated by the surface states for a semiconducting case, which can be analyzed by the Hikami-Larkin-Nagaoka model for two coupled surfaces in the case of strong spin-orbit interaction. However, in the metallic-like case this analysis fails and additional two-dimensional contributions need to be accounted for. Shubnikov-de Haas oscillations and quantized Hall resistance prove as strong indications for the two-dimensional layered metallic behavior. Temperature-dependent magnetotransport properties of high-quality Bi2Se3 single crystalline exfoliated macro and micro flakes are combined with high resolution transmission electron microscopy and energy-dispersive x-ray spectroscopy, confirming the structure and stoichiometry. Angle-resolved photoemission spectroscopy proves a single-Dirac-cone surface state and a well-defined bulk band gap in topological insulating state. Spatially resolved core-level photoelectron microscopy demonstrates the surface stability.


The neuronal channelopathies.
This review addresses the molecular and cellular mechanisms of diseases caused by inherited mutations of ion channels in neurones. Among important recent advances is the elucidation of several dominantly inherited epilepsies caused by mutations of both voltage-gated and ligand-gated ion channels. The neuronal channelopathies show evidence of phenotypic convergence; notably, episodic ataxia can be caused by mutations of either calcium or potassium channels. The channelopathies also show evidence of phenotypic divergence; for instance, different mutations of the same calcium channel gene are associated with familial hemiplegic migraine, episodic or progressive ataxia, coma and epilepsy. Future developments are likely to include the discovery of other ion channel genes associated with inherited and sporadic CNS disorders. The full range of manifestations of inherited ion channel mutations remains to be established.


Comparison of intradialytic hemodynamic tolerance between on-line hemodiafiltration and acetate-free biofiltration with profiled potassium dialysate concentration.
Intradialytic hypotensive episodes are deleterious for hemodialysis (HD) patients. Acetate-free biofiltration with profiled potassium (AFBK) dialysate concentration may improve their cardiovascular stability. The aim of the present crossover study was to compare intradialytic hemodynamic tolerance and biological parameters between online hemodiafiltration (olHDF) and AFBK. Ten frail HD patients (8 males) with a mean age of 66.71- ± 12.31 years were studied for three months on olHDF and AFBK. There was a significant reduction of the hypotensive episodes during the AFBK period compared to the olHDF period. Mean intradialytic systolic and diastolic blood pressures were significantly higher during the AFBK period. There was a significant postdialytic increase in serum sodium concentration with the AFBK compared to olHDF. The dry weight and ultrafiltration indices were significantly higher, and the Kt/V was significantly lower during the AFBK period. Serum albumin concentration significantly increased during the AFBK period. AFBK leads to a significantly improved intradialytic tolerance in hemodynamically instable HD patients.


Molecular and biochemical changes in Locusta migratoria (Orthoptera: Acrididae) infected with Paranosema locustae.
Microsporidia are a group of eukaryotic intracellular parasitic organisms that infect almost all vertebrates and invertebrates. Paranosema locustae are specialized parasites of Orthoptera that are often used as biological controls of locusts, with slow effects of action. In this study, we found that after infection with P. locustae, changes in energy metabolism in male and female Locusta migratoria as were consistent, with no gender differences. During the first 8 days of infection, L. migratoria used sugar as a source of energy. After 8 days, lipids and proteins were consumed to provide energy when the spore load was considerably heavy, and energy supply was insufficient. With increasing infection concentration and time, energy conversion from sugar, fats, and proteins was improved, which may explain why high mortality did not occur until about 15 days after P. locustae infection. The tandem mass tag-based quantitative proteomics analysis revealed that most altered metabolism-related proteins were upregulated (27 of 29 in the metabolic pathway). This result suggests that P. locustae infection accelerated metabolism in L. migratoria, which facilitated the pathogen's life cycle, inhibiting the growth and development of the locusts and eventually killing them. Our findings will be useful to better understand of the chronic pathogenic mechanisms of P. locustae and inform on applications of P. locustae to control locusts.


The sequenced treatment alternatives to relieve depression (STAR*D) trial: a review.
The Sequenced Treatment Alternatives to Relieve Depression (STAR*D) trial is the largest open-label, pragmatic trial that has been undertaken to examine the treatment of major depressive disorder. At a cost of US$35 million over 6 years, STAR*D sought to test the effectiveness both of pharmacotherapy and of cognitive therapy, and to ascertain whether certain treatments are more optimal after one or more failed trials. Patients (n = 2876) who presented to either a psychiatry or family practice setting seeking treatment for depression were included in the STAR*D analysis. In the 4 levels of STAR*D, patients were randomized to various treatment monotherapies, combinations, or augmentation strategies. The primary outcome was remission, based on the Hamilton Depression Rating Scale. Secondary outcomes were response, as measured by clinician and patient self-report as well as various measures of patients' level of function and (or) quality of life. Remission rates for treatment levels 1 to 2 and 3 to 4 were 18% to 30% and 7% to 25%, respectively. There was no difference in effectiveness between any treatments at any treatment level. Patients with longer index episodes, more concurrent psychiatric or general medical disorders, and (or) lower measures of baseline function were less likely to achieve remission. There were no major differences between outcomes in patients treated in primary, compared with specialist care, nor were there significant differences between depression rating scores obtained through clinician ratings, compared with self-report. Results of the STAR*D trial have shed important light on the effectiveness of current treatment strategies for patients with depression.


Effects of high-dose fentanyl on fluid and vasopressor requirements after cardiac surgery.
The effects of two different anesthetic techniques on postoperative cardiopulmonary events, fluid and vasopressor requirements, and overall intensive care unit (ICU) course were studied in patients undergoing elective myocardial revascularization (CABG) (N = 20) or single cardiac valve replacement (N = 20). Patients were randomized to receive either high-dose fentanyl (F, 75 microg/kg followed by 0.3 microg/kg/min) or diazepam-ketamine (0.8 mg/kg D and 2 mg/kg K followed by 0.07 mg/kg/h D and 1 mg/kg/h K). No significant differences in preoperative demographics, prebypass hemodynamics, ischemic crossclamp, or total cardiopulmonary bypass times were noted. Patients receiving F had lower systemic arterial pressures and vascular resistances and required more vasopressors during the first 12 hours postoperatively. They also had a more positive cumulative fluid balance at 24 and 48 hours postoperatively, despite a higher incidence of postoperative diuretic use in CABG patients receiving F. Rectal temperature was significantly higher at four and eight hours postoperatively in F patients. Time until arousal and length of ICU stay were significantly greater with F, although duration of intubation did not differ between anesthetic techniques. Although the data does not provide an explanation for these differences, it indicates that these two techniques produce quite different physiologic responses in the postoperative period following cardiac surgery. In summary, it was found that F for cardiac surgery was accompanied by increased postoperative fluid and vasopressor requirements with increased Qsp/Qt and longer ICU stays compared to diazepam and ketamine. This study suggests that some anesthetic techniques may provide less complicated and less costly postoperative courses than others, with the same outcome after cardiac surgery. Information on the postoperative effects of long-acting anesthetic agents is sparse and more studies of these effects and their mechanisms are necessary because they may affect patient management after cardiac surgery.


Diesel exhaust particle exposure in vitro impacts T lymphocyte phenotype and function.
Diesel exhaust particles (DEP) are major constituents of ambient air pollution and their adverse health effect is an area of intensive investigations. With respect to the immune system, DEP have attracted significant research attention as a factor that could influence allergic diseases interfering with cytokine production and chemokine expression. With this exception, scant data are available on the impact of DEP on lymphocyte homeostasis. Here, the effects of nanoparticles from Euro 4 (E4) and Euro 5 (E5) light duty diesel engines on the phenotype and function of T lymphocytes from healthy donors were evaluated. T lymphocytes were isolated from peripheral blood obtained from healthy volunteers and subsequently stimulated with different concentration (from 0.15 to 60 μg/ml) and at different time points (from 24 h to 9 days) of either E4 or E5 particles. Immunological parameters, including apoptosis, autophagy, proliferation levels, mitochondrial function, expression of activation markers and cytokine production were evaluated by cellular and molecular analyses. DEP exposure caused a pronounced autophagic-lysosomal blockade, thus interfering with a key mechanism involved in the maintaining of T cell homeostasis. Moreover, DEP decreased mitochondrial membrane potential but, unexpectedly, this effect did not result in changes of the apoptosis and/or necrosis levels, as well as of intracellular content of adenosine triphosphate (ATP). Finally, a down-regulation of the expression of the alpha chain of the interleukin (IL)-2 receptor (i.e., the CD25 molecule) as well as an abnormal Th1 cytokine expression profile (i.e., a decrease of IL-2 and interferon (IFN)-γ production) were observed after DEP exposure. No differences between the two compounds were detected in all studied parameters. Overall, our data identify functional and phenotypic T lymphocyte parameters as relevant targets for DEP cytotoxicity, whose impairment could be detrimental, at least in the long run, for human health, favouring the development or the progression of diseases such as autoimmunity and cancer.


Evaluation of daunorubicin (NSC-82151) and methotrexate (NSC-740) in combination as a remission maintenance regimen in the treatment of acute leukemia.
The addition of a single monthy dose of daunorubicin (25 mg/m given intravenously) to the maximum tolerated dose of methotrexate (25 mg/m/dose given orally twice weekly) was evaluated as a maintenance regimen in 0 children with acute lymphoblastic leukemia. Seven patients who were nonevaluable were excluded from the study; 31 patients received methotrexate alone and 32 patients received a combination of methotrezate and daunorubicin. There was no significant difference between the distribution of remission times for the two maintenance regimens (medians of 147 and 162 days, respectively). For patients with only one previous remission, the median length of remission in this study was significantly shorter than that of the original remission time. Despite the known myelosuppressive effects of both agents, daunorubicin did not cause additional toxicity nor did it interfere with the scheduled methotrexate therapy. Although the addition of daunorubicin to methotrexate does not appear to be beneficial in the treatment of acute leukemia, this combination could be evaluated in other cancers in which both agents are known to be effective.


Chiral Nematic Liquid Crystal Behavior of Core-Shell Hybrid Rods Consisting of Chiral Cellulose Nanocrystals Dressed with Non-chiral Conformal Polymeric Skins.
The current work investigates how the nanoscale conformal coating layers of non-chiral polymeric materials can influence the chiral nematic liquid crystal (CLC) behaviors of the rodlike cellulose nanocrystals (CNCs), the bio-derived nanomaterials that have attracted significant attention. For this, we developed strategies to coat the CNC rods on the single-particle level with a homogeneous bioinspired polydopamine (PDA) layer, leading to well-defined core-shell CNC@PDA rods with various PDA coating thicknesses and excellent colloidal stability. Comprehensive investigation revealed that the CNC@PDA hybrid nanorods in concentrated suspensions form well-defined nematic liquid crystal phases with clear phase separation behavior that depend on the rod concentrations and ionic strengths, typical of charged rods. Most intriguingly, the nematic LC phases formed by the CNC@PDA rods with the PDA coating thickness achieved herein are indeed the perfect CLC phases, which form following the classic pathway of nucleation and coalesce of chiral tactoids and have colorful chiral fingerprints standing out from the dark suspensions. The pitches of the CLC phase increase sharply with increasing PDA coating thicknesses and are significantly larger than those of the pristine CNCs. Such observations can be attributed to the blurring effects of the PDA coating on the intrinsic surface chiral features of CNC of whatever origins that drive the formation of the CLC phases, resulting in weakening chiral interactions between CNC@PDA rods. Besides benefiting the understanding of the long-sought origin of the CLC phases of the pristine CNC, the current work demonstrates the possibility of controlling the CLC phase behaviors of CNC by tuning the thickness of the coating materials and also serves as the first example of directly transferring the unique chirality of CNC to other non-chiral materials.


Cyclic stretch of human lung cells induces an acidification and promotes bacterial growth.
The reasons for bacterial proliferation in the lungs of mechanically ventilated patients are poorly understood. We hypothesized that prolonged cyclic stretch of lung cells influenced bacterial growth. Human alveolar type II-like A549 cells were submitted in vitro to prolonged cyclic stretch. Bacteria were cultured in conditioned supernatants from cells submitted to stretch and from control static cells. Escherichia coli had a marked growth advantage in conditioned supernatants from stretched A549 cells, but also from stretched human bronchial BEAS-2B cells, human MRC-5 fibroblasts, and murine RAW 264.7 macrophages. Stretched cells compared with control static cells acidified the milieu by producing increased amounts of lactic acid. Alkalinization of supernatants from stretched cells blocked E. coli growth. In contrast, acidification of supernatants from control cells stimulated bacterial growth. The effect of various pharmacological inhibitors of metabolic pathways was tested in this system. Treatment of A549 cells and murine RAW 264.7 macrophages with the Na(+)/K(+)-ATPase pump inhibitor ouabain during cyclic stretch blocked both the acidification of the milieu and bacterial growth. Several pathogenic bacteria originating from lungs of patients with ventilator-associated pneumonia (VAP) also grow better in vitro at slightly acidic pH (pH 6-7.2), pH similar to those measured in the airways from ventilated patients. This novel metabolic pathway stimulated by cyclic stretch may represent an important pathogenic mechanism of VAP. Alkalinization of the airways may represent a promising preventive strategy in ventilated critically ill patients.


Overview of clinical trials on periodontal regeneration.
Periodontal regeneration means healing after periodontal surgery that results in the restoration of the tooth-supporting tissues; namely, cementum, alveolar bone, and periodontal ligament. The response of the periodontium to conventional periodontal flap surgery is primarily repair. Regenerative flap procedures involve modifications intended to alter variables in the normal wound healing response to shift the outcome from repair to regeneration. Technical manipulations, devices, pharmacological agents, and biologicals have been used to obtain regeneration. Evaluation of periodontal therapy is done by clinical and histological assessments. Human clinical trials depend on probing, re-entry, and radiographic measurements which cannot distinguish between periodontal repair and regeneration. Histological studies are difficult in humans because they require the retrieval of tissue blocks containing the teeth and their surrounding tissues, and for this reason these studies have largely been done in animal models; however, the use of animal models creates problems of interpretation when applying the information to the human. There are other fundamental problems related to assessment of new methods of periodontal regeneration. Ideally, periodontal regeneration is attempted after resolving the periodontitis that produced the loss of periodontal support. Does the type of periodontal disease affect the outcome? If it does, then separate clinical trials would be indicated to manage defects produced by different diseases. Furthermore, certain types of periodontal defects apparently respond better than others (for example, intrabony defects versus suprabony defects). Does each type of defect require a separate trial? Do subjects respond differently and, if so, how should this be factored into selection of the study population and sample size? There appear to be more questions than answers concerning the design of clinical trials for periodontal regeneration. This is not surprising since regeneration has been possible for only a few years, is apparently unpredictable, and the origin and nature of the new tissue are poorly understood. These factors make it important that the design of clinical trials be openly and thoroughly discussed so that practical solutions are forthcoming.


Safe "cloudification" of large images through picker APIs.
The "Box model" allows users with no particular training in informatics, or access to specialized infrastructure, operate generic cloud computing resources through a temporary URI dereferencing mechanism known as "drop-file-picker API" ("picker API" for sort). This application programming interface (API) was popularized in the web app development community by DropBox, and is now a consumer-facing feature of all major cloud computing platforms such as Box.com, Google Drive and Amazon S3. This reports describes a prototype web service application that uses picker APIs to expose a new, "cloudified", API tailored for image analysis, without compromising the private governance of the data exposed. In order to better understand this cross-platform cloud computing landscape, we first measured the time for both transfer and traversing of large image files generated by whole slide imaging (WSI) in Digital Pathology. The verification that there is extensive interconnectivity between cloud resources let to the development of a prototype software application that exposes an image-traversing REST API to image files stored in any of the consumer-facing "boxes". In summary, an image file can be upload/synchronized into a any cloud resource with a file picker API and the prototype service described here will expose an HTTP REST API that remains within the safety of the user's own governance. The open source prototype is publicly available at sbu-bmi.github.io/imagebox. Availability The accompanying prototype application is made publicly available, fully functional, with open source, at http://sbu-bmi.github.io/imagebox://sbu-bmi.github.io/imagebox. An illustrative webcasted use of this Web App is included with the project codebase at https://github.com/SBU-BMI/imageboxs://github.com/SBU-BMI/imagebox.


[Cloning of Brucella genes in Escherichia coli K12 cells and analysis of products of the cloned genes].
The libraries of Brucella melitensis 565 and Brucella abortus 99 in Escherichia coli cells have been constructed. Some clones of Escherichia coli producing the specific brucella antigens have been found in immunological tests with brucella antiserum. Two strains producing antigens have been characterized, one being from Brucella melitensis 565 and another from Brucella abortus 99 clone libraries . Both strains synthesize two antigens that were studied by immunoelectrophoresis, immunoblotting after treatment of antigen preparations with different physical and chemical agents substrate specific enzymes. Both strains are found to synthesize the specific brucella antigens of protein nature. One of them has the mol mass about 15 kD, another--31-32 kD. The 31-32 kD antigen can be, evidently, referred to as the main protein of an outer membrane of brucella.


New source of genetic polymorphisms in Lepidoptera?
The variability level of the ISSR (inter-simple sequences repeat) primer (GACA)4 was examined in the three Lepidoptera families Pyralidae, Sphingidae and Pieridae. Our study shows that the tetra-repeat (GACA)n is evidently present in sufficient numbers in these butterflies to provide informative DNA fingerprints. The variability is mostly rather high, but within a comparable range to other ISSR studies. Although less polymorphisms may be encountered in some butterfly families, this study indicates that high variability of this marker may be a common characteristic of Lepidoptera genomes. An appeal for a minimal level of standardization of ISSR-PCR data analysis is formulated to enable an exact comparison between the groups of organisms studied with this fingerprint technique.


Tics are caused by alterations in prefrontal areas, thalamus and putamen, while changes in the cingulate gyrus reflect secondary compensatory mechanisms.
Despite strong evidence that the pathophysiology of Tourette syndrome (TS) involves structural and functional disturbances of the basal ganglia and cortical frontal areas, findings from in vivo imaging studies have provided conflicting results. In this study we used whole brain diffusion tensor imaging (DTI) to investigate the microstructural integrity of white matter pathways and brain tissue in 19 unmedicated, adult, male patients with TS "only" (without comorbid psychiatric disorders) and 20 age- and sex-matched control subjects. Compared to normal controls, TS patients showed a decrease in the fractional anisotropy index (FA) bilaterally in the medial frontal gyrus, the pars opercularis of the left inferior frontal gyrus, the middle occipital gyrus, the right cingulate gyrus, and the medial premotor cortex. Increased apparent diffusion coefficient (ADC) maps were detected in the left cingulate gyrus, prefrontal areas, left precentral gyrus, and left putamen. There was a negative correlation between tic severity and FA values in the left superior frontal gyrus, medial frontal gyrus bilaterally, cingulate gyrus bilaterally, and ventral posterior lateral nucleus of the right thalamus, and a positive correlation in the body of the corpus callosum, left thalamus, right superior temporal gyrus, and left parahippocampal gyrus. There was also a positive correlation between regional ADC values and tic severity in the left cingulate gyrus, putamen bilaterally, medial frontal gyrus bilaterally, left precentral gyrus, and ventral anterior nucleus of the left thalamus. Our results confirm prior studies suggesting that tics are caused by alterations in prefrontal areas, thalamus and putamen, while changes in the cingulate gyrus seem to reflect secondary compensatory mechanisms. Due to the study design, influences from comorbidities, gender, medication and age can be excluded.


Expertise Modulates Students' Perception of Pain From a Self-Perspective: Quasi-Experimental Study.
Perception of stimuli presented in a virtual dentistry environment affects regions of the brain that are related to pain perception. We investigated whether neural correlates of virtual pain perception are affected by education in dentistry. In this functional magnetic resonance imaging study, a sample of 20 dental students and 20 age-matched controls viewed and listened to video clips presenting a dental treatment from the first-person perspective. An anxiety questionnaire was used to assess the level of dental anxiety. Neural correlates of pain perception were investigated through classic general linear model analysis and in-house classification methods. Dental students and naïve controls exhibited similar anxiety levels for invasive stimuli. Invasive dentistry scenes evoked a less affective component of pain in dental students compared with naïve controls (P<.001). Reduced affective pain perception went along with suppressed brain activity in pain matrix regions including the insula, anterior cingulate cortex, and basal ganglia. Furthermore, a substantial reduction of brain activity was observed in motor-related regions, particularly the supplementary motor area, premotor cortex, and basal ganglia. Within this context, a classifier analysis based on neural activity in the nucleus lentiformis could identify dental students and controls on the individual subject level in 85% of the cases (34 out of 40 participants, sensitivity=90%, specificity=80%). Virtual dental treatment activates pain-related brain regions in controls. By contrast, dental students suppress affective and motor-related aspects of pain. We speculate that dental students learn to control motoric aspects of pain perception during their education because it is a prerequisite for the professional manual treatment of patients. We discuss that a specific set of learning mechanisms might affect perceived self-efficacy of dental students, which in turn might reduce their affective component of pain perception.


Behavioral differences among fourteen inbred mouse strains commonly used as disease models.
We compared the behavior of 14 inbred mouse strains and an F1 hybrid commonly used in transgenic and knockout production. These strains were 129P3/J, 129S1/SvImJ, 129S6/SvEvTac, 129T2/SvEmsJ, 129X1/SvJ (formerly 129/J, 129/Sv-p+Tyr+Kitl+/J, 129/SvEvTac, 129SvEmsJ, and 129/SvJ, respectively), A/JCrTac, BALB/cAnNTac, C3H/HeNTac, C57BL/6J, C57BL/6NTac, DBA/2NTac, FVB/NTac, NOD/MrkTac, SJL/JCrNTac, and the hybrid B6129S6F1Tac. Performance in three behavioral tests (rotorod, open-field activity-habituation, and contextual and cued fear conditioning) was determined. On the rotorod assay, SJL/JCrNTac mice had the shortest latencies to fall on the first day of testing, and DBA/2NTac mice showed impaired motor learning. Open-field behavior was analyzed using the parameters total distance, center distance, velocity, and vertical activity. 129T2/EvEmsJ and A/JCrTac were least active in the open field, whereas NOD/MrkTac mice were most active. Contrary to earlier studies, we found that all strains habituated to the open field in at least one of these parameters. In contextual and cued fear conditioning, all strains displayed activity suppression. However, FVB/NTac mice reacted less strongly to both context and cue than did most of the other strains. There were no significant behavioral differences between C57BL/6J and C57BL/6NTac, except for higher open-field activity in C57BL/6J female mice. These findings illustrate the importance of the appropriate selection of background strain for transgenic, gene targeting, or drug research.


[Analysis of intracellular proapoptotic (Bax, Bak) and antiapoptotic (Bcl-2, Bcl-XL) proteins expression in thyrocytes from young patients with immune and non-immune thyroid disorders].
Apoptosis one of the form of programmed cell death is a physiological occurrence, requisite to the correct function of every organism. This is an active process that proceeds with a participation of the cellular metabolism embracing the activation of genes and the synthesis of proteins. The signal to apoptosis can be started practically in every cell of our organism. Disturbances of the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders. The aim of this study was to estimate the expression of proapoptotic (Bax, Bak) and antiapoptotic (Bcl-2, Bcl-XL) proteins in thyroid tissues from 12 patients with Graves' disease (GD), 10 with non-toxic nodular goitre (NTNG) and 10 with toxic nodular goitre (TNG). Criteria for qualification of Graves' patients: large goitre, ophthalmopathy, TRAb > 5 U/L, positive titre of anti-TPO and anti-TG antibodies and concentration of TSH <0.45 microIU/mL more the 2-3 months from onset of the disease. Detection of apoptotic proteins in thyroid follicular cells was performed by Western Blot. These analysis was confirmed by immunohistochemistry using monoclonal antibodies in DAB chromogene visuality and marked by Mayer's haematoxylin. Identification of antiapoptotic Bcl-2 and Bcl-XL molecules in the thyroid follicular cells revealed a higher expression of both proteins in patients with Graves' disease (+++; ++, respectively) in comparison to patients with NTNG (++/+; +) and TNG (++; +). The detection of proapoptotic molecules showed higher expression of Bak (++/+) and Bax (+) in Graves' thyroid tissues while Bax was in trace amount in NTNG (0/+) and TNG (0/+). We conclude that alteration in the expression of antiapoptotic and proapoptotic proteins on surface of thyroid follicular cells may play a role in the pathogenesis of thyroid autoimmune disorders. In addition, suppression of apoptosis in Graves' disease led to predominance for proliferation of thyroid follicular cells which is responsible for goitre formation.


Impact of Prior Treatment on the Efficacy of Adoptive Transfer of Tumor-Infiltrating Lymphocytes in Patients with Metastatic Melanoma.
Adoptive cell transfer (ACT) of autologous tumor-infiltrating lymphocytes (TIL) can mediate durable responses in patients with metastatic melanoma. This retrospective analysis provides long-term follow-up and describes the effect of prior therapy on outcomes after ACT-TIL. Patients with metastatic melanoma underwent surgical resection of a tumor for generation of TILs and were treated with a lymphodepleting preparative regimen followed by adoptive transfer of TILs and intravenous IL2. Clinical characteristics of enrolled patients and treatment characteristics of TIL infusion products over two decades of ACT were analyzed to identify predictors of objective response. Adoptive transfer of TILs mediated an objective response rate of 56% (108/192) and median melanoma-specific survival of 28.5 months in patients naïve to anti-programmed cell death-1 (PD-1) therapy compared with 24% (8/34) and 11.6 months in patients refractory to anti-PD-1 (aPD-1). Among patients with BRAF V600E/K-mutated disease, prior treatment with targeted molecular therapy was also associated with a decreased response rate (21% vs. 60%) and decreased survival (9.3 vs. 50.7 months) when compared with those patients naïve to targeted therapy. With a median potential follow-up of 89 months, 46 of 48 complete responders in the aPD-1-naïve cohort have ongoing responses after a single treatment and 10-year melanoma-specific survival of 96%. Patients previously treated with PD-1 or MAPK inhibition are significantly less likely to develop durable objective responses to ACT-TIL. While ACT-TIL is currently being investigated for treatment-refractory patients, it should also be considered as an initial treatment option for eligible patients with metastatic melanoma. See related commentary by Sznol, p. 5156.


Prognostic role of non-sustained ventricular tachycardia in a large cohort of patients with idiopathic dilated cardiomyopathy.
The identification of patients with idiopathic dilated cardiomyopathy (IDC) at higher risk of sudden death (SD) is still an unsolved issue, and the role of non-sustained ventricular tachycardia (NSVT) uncertain. The effect of NSVT on total mortality, SD and life-threatening arrhythmias was evaluated in 554 patients with IDC on optimal medical treatment and at long-term follow-up (81 +/- 58 months). At diagnosis, 240 patients (43%) had NSVT at Holter monitoring and 314 (57%) did not. During follow-up, 189 patients (5/100 patients-year) died or underwent heart transplantation; SD occurred in 53 patients (1.4/100 patients-year); SD + non-fatal ventricular arrhythmias occurred in 75 patients (2/100 patients-year). Patients with and without NSVT at diagnosis had the same 5-year transplant-free survival rate (76 vs 76%, p = NS) and a similar incidence of SD (10 vs 7%, p = NS). The length and rate of NSVT did not show any significant relationship with the outcome. Only heart failure symptoms (NYHA class III-IV) (hazard ratio [HR] 1.9, p = 0.015) and severe left ventricular impairment (left ventricular ejection fraction < or = 0.30 and left ventricular end-diastolic diameter > or = 70 mm) (HR 2.7, p < 0.0001) were independently associated with higher SD risk. At multivariate analysis the presence of frequent NSVT episodes (> or = 3 runs/day) was associated with an increased risk of total mortality (HR 1.68, p = 0.041) and of major ventricular arrhythmias (HR 2.11, p = 0.037), but only in the subgroup of patients with severe left ventricular impairment. Patients with advanced heart failure symptoms, severe left ventricular dysfunction and dilation had a higher risk of SD independently of NSVT. The finding of more frequent NSVT was associated with an increased risk of all-cause mortality and of major ventricular arrhythmias in patients with severe left ventricular impairment.


Cyclical konzo epidemics and climate variability.
Konzo epidemics have occurred during droughts in the Democratic Republic of Congo (DR Congo) for >70 years, but also in Mozambique, Tanzania, and the Central African Republic. The illness is attributed to exposure to cyanide from cassava foods, on which the population depends almost exclusively during droughts. Production of cassava, a drought-resistant crop, has been shown to correlate with cyclical changes in precipitation in konzo-affected countries. Here we review the epidemiology of konzo as well as models of its pathogenesis. A spectral analysis of precipitation and konzo is performed to determine whether konzo epidemics are cyclical and whether there is spectral coherence. Time series of environmental temperature, precipitation, and konzo show cyclical changes. Periodicities of dominant frequencies in the spectra of precipitation and konzo range from 3 to 6 years in DR Congo. There is coherence of the spectra of precipitation and konzo. The magnitude squared coherence of 0.9 indicates a strong relationship between variability of climate and konzo epidemics. Thus, it appears that low precipitation phases of climate variability reduce the yield of food crops except cassava, upon which the population depends for supply of calories during droughts. Presence of very high concentrations of thiocyanate (SCN(-) ), the major metabolite of cyanide, in the bodily fluids of konzo subjects is a consequence of dietary exposure to cyanide, which follows intake of poorly processed cassava roots. Because cyanogens and minor metabolites of cyanide have not induced konzo-like illnesses, SCN(-) remains the most likely neurotoxicant of konzo. Public health control of konzo will require food and water programs during droughts. [Correction added on 26 February 2015, after first online publication: abstract reformatted per journal style]


Genetic structure of Sakhalin spruce (Picea glehnii) in northern Japan and adjacent regions revealed by nuclear microsatellites and mitochondrial gene sequences.
The genetic structure of Sakhalin spruce (Picea glehnii) was studied across the natural range of the species, including two small isolated populations in south Sakhalin and Hayachine, by using six microsatellite loci and maternally inherited mitochondrial gene sequences. We also analyzed P. jezoensis, a sympatric spruce in the range. Genetic diversity of P. glehnii was higher in central Hokkaido and the lowest in the Hayachine. Bayesian clustering and principal coordinate analysis by using the microsatellites indicated that the Hayachine was clearly distinct from other populations, implying that it had undergone strong genetic drift since the last glacial period. P. glehnii harbored four mitochondrial haplotypes, two of which were shared with P. jezoensis. One of the two was observed without geographical concentration, suggesting its derivation from ancestral polymorphism. Another was observed in south Sakhalin and in P. jezoensis across Sakhalin. The Bayesian clustering--by using four microsatellite loci, including P. jezoensis populations--indicated unambiguous species delimitation, but with possible admixture of P. jezoensis genes into P. glehnii in south Sakhalin, where P. glehnii is abundantly overwhelmed by P. jezoensis; this might explain the occurrence of introgression of the haplotype of P. jezoensis into P. glehnii.


[Recent advances in the pathophysiology of atrial fibrillation].
This review is a summary of the pathophysiology of atrial fibrillation (AF) and the progress in the understanding of this arrhythmia. The following factors should be considered in the genesis and maintenance of AF. The genetic factor is involved in cases of familial AF. Predisposing structural factors: atrial dilation, structural feature that allows the development of AF. Predisposing structural factors: the role of the Bachmann's bundle and frequency gradients between the left and right atrium. Electrophysiological factors: heterogeneous refractory periods favor the fibrillatory conduction. Triggers: abnormal electrical activity (ectopic foci). Modulating factors: autonomic nervous system. Increased vagal tone shortens atrial refractory periods, creating a greater dispersion of the refractory periods and the generation of reentries in the context of triggered activity. Finally, there are three types of atrial remodeling secondary to AF: structural, contractile, electrical. They are interrelated and contribute to maintaining the AF ("AF begets AF").


Ultra-Sensitive HIV-1 Latency Viral Outgrowth Assays Using Humanized Mice.
In the current quest for a complete cure for HIV/AIDS, highly sensitive HIV-1 latency detection methods are critical to verify full viral eradication. Until now, the in vitro quantitative viral outgrowth assays (qVOA) have been the gold standard for assessing latent HIV-1 viral burden. However, these assays have been inadequate in detecting the presence of ultralow levels of latent virus in a number of patients who were initially thought to have been cured, but eventually showed viral rebound. In this context, new approaches utilizing in vivo mouse-based VOAs are promising. In the murine VOA (mVOA), large numbers of CD4+ T cells or PBMC from aviremic subjects are xenografted into immunodeficient NSG mice, whereas in the humanized mouse-based VOA (hmVOA) patient CD4+ T cell samples are injected into BLT or hu-hematopoetic stem cells (hu-HSC) humanized mice. While latent virus could be recovered in both of these systems, the hmVOA provides higher sensitivity than the mVOA using a fewer number of input cells. In contrast to the mVOA, the hmVOA provides a broader spectrum of highly susceptible HIV-1 target cells and enables newly engrafted cells to home into preformed human lymphoid organs where they can infect cells in situ after viral activation. Hu-mice also allow for both xenograft- and allograft-driven cell expansions with less severe GvH providing a longer time frame for potential viral outgrowth from cells with a delayed latent viral activation. Based on these advantages, the hmVOA has great potential in playing an important role in HIV-1 latency and cure research.


The meaning of occupation, occupational need, and occupational therapy in a military context.
Despite occupational therapists having strong historical ties to the Canadian military, there are currently no uniformed occupational therapists and only a few permanent occupational therapists employed by Canadian Forces. Occupational therapy is provided, in the main, through civilian occupational therapists. Occupational therapists have unique skills that can contribute to the existing Canadian Forces Physical Medicine and Rehabilitation Services Department. To establish the depth and scope of their work, this article explains the theoretical underpinnings of occupational therapy. Examples are provided of possible occupational therapy for populations of Canadian Forces members: (1) those with transient, intermittent injuries; (2) those returning from overseas missions with very serious injuries or severe injuries; and (3) those with permanent injuries who are transitioning from the Canadian Forces into the civilian workforce. Interventions for mental health issues are interwoven with those targeting physical issues. The article suggests that occupational therapists employed on a permanent basis by the Canadian Forces can contribute in a more comprehensive manner to the wider rehabilitation of Canadian Forces members. The article has applicability to occupational therapy military services in other countries.


Ferritin-templated synthesis and self-assembly of Pt nanoparticles on a monolithic porous graphene network for electrocatalysis in fuel cells.
The monolithic three-dimensional (3D) graphene network is used as the support for Pt nanoparticles (NPs) to fabricate an advanced 3D graphene-based electrocatalyst. Distinct from previous strategies, the monodispersed Pt NPs with ultrafine particle size (∼3 nm) are synthesized using ferritin protein nanocages as the template and subsequently self-assembled on the 3D graphene by leveraging on the hydrophobic interaction between the ferritin and the graphene. Following the self-assembly, the ferritins are removed, resulting in a stable Pt NP/3D graphene composite. The composite exhibits much enhanced electrocatalytic activity for methanol oxidation as compared with both Pt NP/chemically reduced graphene oxide (Pt/r-GO) and state-of-the-art Pt/C catalyst. The observed electrocatalytic activity also shows marked improvement over Pt/3D graphene prepared by pulse electrodeposition of Pt. This study demonstrates that protein nanocage templating and assembly are promising strategies for the fabrication of functional composites in catalysis and fuel cell applications.


Density functional calculations of the properties of silicon-substituted hydroxyapatite.
Ab initio density functional plane-wave calculations are performed on silicon-substituted hydroxyapatite (SiHA). Formation energies are obtained for the substitution of a phosphorus atom by a silicon atom in each of the six phosphate groups of the unit cell in turn. It is found that the co-removal of a hydroxyl group to maintain charge neutrality is energetically favourable and the calculated unit cell volumes for the single silicon substitutions agree extremely well with experimental observation. The substitution of a second silicon atom in the unit cell is found to be almost as energetically favourable as the first (and on one site more favourable) and there can be an attractive interaction between the two Si substituents when they are closely separated. However, experimental observation suggests that for this concentration of silicon a phase transformation to a different structure occurs which, because of the imposed boundary conditions, could not be accessed in the calculations. The density of states of the SiHA indicates that new states are introduced deep into the valence band and the band gap decreases by 1.6 eV compared to phase-pure HA. No new states are introduced into the band gap indicating that the Si incorporation does not make the material inherently electrically active. Furthermore a population analysis shows that the Si impurity has only a small effect on the neighbouring ionic charge.


Enhanced geographically restricted surveillance simulates sporadic Creutzfeldt-Jakob disease cluster.
Spatio-temporal clustering of sporadic Creutzfeldt-Jakob disease (sCJD) has been recognized and investigated previously in various global settings including Australia. Generally, despite often extensive investigation, explanations such as point source outbreaks and plausible case-to-case transmission links have not been identified to explain the apparently higher case rates than expected. In the context of national surveillance during the period 1993-2006, an increased number of cases of sCJD were recognized in a circumscribed coastal region of eastern Australia. To assess the significance of this apparent clustering, the Spatial Scan Statistic was used to examine for geographic excess of CJD mortality at spatial and temporal combined, spatial only and temporal only levels. A significant spatial cluster was confirmed, encompassing three contiguous statistical local areas within the state of New South Wales (NSW). Detailed epidemiological analysis did not reveal a plausible cross-over or point source transmission event. Further evaluation prompted the conclusion that vigilant and motivated managing clinicians in this geographically circumscribed area of NSW evinced a sustained higher level of clinical awareness for the broad phenotypic spectrum of CJD with reliable referral of suspect cases for further investigation. In addition, these physicians established and maintained a well-coordinated and active approach to suspect CJD autopsy. This combination of factors translated into a higher intensity of surveillance at approximately twice the rate per population observed in the entire state, culminating in twice the incidence of sCJD at around 2.28 cases/million population/year. The hypothesis that intensity of surveillance for rare disorders can be objectively measured and that this can positively correlate with disease incidence deserves further exploration. It may prove to be an important insight into the varying incidence rates over periods of time within individual nations and between different countries.


[Insecticide resistance of Bemisia tabaci field populations].
Resistance to 13 insecticides in field populations of Bemisia tabaci from six regions (Fuzhou, Zhangzhou, Longyan, Sanming, Nanping, Ningde) of Fujian Province, China was monitored by adult leaf-dipping bioassay. Compared with the susceptible SUD-S strain, all the six field populations exhibited high levels of resistance to lambda-cyhalothrin (838.38-2460.52 fold), fenpropathrin (244.64-834.29 fold), cypermethrin (116.02-266.35 fold), deltamethrin (81.75-124.18 fold), acephate (425.18-875.56 fold) and chlorpyrifos (54.53-78.43 fold), moderate levels of resistance to dimethoate (14.16-17.66 fold), low to moderate levels of resistance to dichlorvos (6.23-11.25 fold) and low levels of resistance to methomyl (4.07-5.66 fold), respectively. Among these six field-collected populations, only Zhangzhou population had moderate resistance to imidacloprid, acetamiprid and thiamethoxam (23.08 fold, 10.32 fold and 24.60 fold, respectively). All field strains tested displayed no resistance to abamectin.


STM study of molecule double-rows in mixed self-assembled monolayers of alkanethiols.
Using scanning tunnelling microscopy (STM), we have studied mixed self-assembled monolayers of linear alkanethiol molecules. Nonanedithiol (C9S2), nonanethiol (C9S), decanethiol (C10S), and dodecanethiol (C12S) were inserted into a self-assembled octanethiol (C8S) host matrix monolayer on an Au(111) surface using a two-step method. Quasi-one-dimensional double-row structures were found in the ordered, close-packed domains of the C8S matrix for each mixed monolayer system. These close-packed domains coexist with ordered striped phase domains (for C9S and C10S) or with a disordered phase (for C9S2 and C12S). Results from high-resolution images suggest that the double-rows are composed of inserted non-nearest-neighbor substitutional molecules, the ordering of which may be a result of locally induced surface stress.


Effects of atmospheric-pressure non-thermal bio-compatible plasma and plasma activated nitric oxide water on cervical cancer cells.
Atmospheric-pressure non-thermal bio-compatible plasma is a partially ionized gas with electrically charged particles. Previous studies demonstrated that dielectric barrier discharge (DBD) plasma could induce apoptosis of various cancer cells, in particular demonstrating the selective cytotoxicity of cancer cells over normal cells. Therefore, DBD plasma can be considered as a potential cancer treatment method for clinical applications. We previously developed a microwave jet plasma system, producing nitric oxide called nitric oxide-plasma activated water (NO-PAW). In this study, we explored the effects of NO-PAW on a cervical cancer cell line, in comparison with DBD plasma. The cytotoxicity results showed that the treatment of HeLa cell with DBD for 4 minutes and 7 μM concentration of NO-PAW could reach almost IC60. For the apoptosis assay, 4 minutes treatment of DBD could induce 7% apoptotic effect, whereas 7 μM NO-PAW could induce 18% apoptotic effect. In addition, we assumed that both DBD plasma and NO-PAW could induce HeLa cell apoptosis by facilitating an accumulation of intracellular reactive oxygen and nitrogen species (RONS). Although further detail on the molecular signal pathway is still needed, DBD and NO-PAW could become promising applications for effective and safe clinical trials for cancer therapy.


Significance of endometrial cells in cervicovaginal smears.
A study was undertaken to determine the significance of finding normal endometrial cells in routine cervicovaginal smears. The results indicate that the presence of normal endometrial cells in cervicovaginal smears in patients under 40 is generally not of significance, but similar findings in older patients correlate significantly with pathologic changes in the endometrium. In the older age group, therefore, such findings clearly indicate a need for further evaluation, including tissue sampling of the endometrium.


An analysis of the efferent connections of the septal area in the cat.
The neuroanatomical organization of the efferent connections of the septal area in the cat was analyzed by the use of anterograde ([3H]leucine radioautography) and retrograde (horseradish peroxidase histochemistry) tracing techniques. The results indicate that the lateral septal nucleus projects to the nuclei of the diagonal band, preoptic area, lateral hypothalamus, and supramammillary region. The projections of the septofimbrial nucleus supply the nuclei of the diagonal band and the medial habenular nucleus. Projection targets of the vertical limb of the diagonal band are widespread and include the preoptic area, lateral hypothalamus, anterior limbic cortex, amygdala, medial habenular nucleus, interpeduncular nucleus and hippocampal formation. The projection from the vertical limb to the hippocampal formation is organized in a topographical manner in such a fashion that cells positioned near the midline project to the dorsal hippocampus and adjoining subicular cortex while fibers originating from cells situated more laterally project to more ventral parts of the hippocampal formation. In general, the projections from the horizontal limb were similar to those from the vertical limb, but several differences were noted. Fibers arising from the horizontal limb are distributed to the ventral tegmental area and interpeduncular nucleus but this region seems to lack a projection to either the habenular complex or to the ventral aspect of the hippocampal formation. Fibers arising from the bed nucleus of the anterior commissure are distributed to the preoptic region, lateral hypothalamus, supramammillary region, posterior aspect of the medial mammillary nucleus and lateral habenular nucleus.


A comparative randomized double-blind clinical trial of isoaminile citrate and chlophedianol hydrochloride as antitussive agents.
The efficacy and safety of a new centrally acting antitussive agent, isoaminile citrate, was compared with that of chlophedianol hydrochloride in a double-blind, randomized interpatient study. A total of 66 patients participated, two and four patients were lost to follow-up with isoaminile and chlophedianol, respectively. In the experimentally induced cough in 12 normal human subjects, isoaminile (40 mg) was as effective as chlophedianol (20 mg), but its duration of action was somewhat longer. One subject developed allergic skin rash with chlophedianol and was withdrawn from the study. In 60 patients with cough associated with chest diseases, isoaminile (40 mg, 3 x daily) was as effective as chlophedianol (20 mg, 3 x daily) in suppressing cough as judged from the 3-h and 24-h cough counts. The increase in PEFR at day 7 of treatment was somewhat more marked with chlophedianol as compared with isoaminile. None of the drugs interfered with the expectoration process. The side effects observed were few, mild in nature, and did not require a decrease in dose or withdrawal of treatment in any of the patients. Isoaminile citrate was concluded to be an effective and relatively safe antitussive agent. Isoaminile citrate, alpha(isopropyl)-alpha-(beta-dimethylaminoproyl) phenylacetonitrile citrate, is a centrally acting antitussive agent. In animal experiments this drug was as efficacious as codeine but was devoid of any respiratory depressant effect [Krause 1958, Kuroda et al. 1971]. This controlled double-randomized interpatient study was designed to test the comparative efficacy and safety of isoaminile and chlophedianol, another centrally acting antitussive, in humans.


Inorganic phosphate homeostasis and the role of dietary phosphorus.
Inorganic phosphate (Pi) is required for cellular function and skeletal mineralization. Serum Pi level is maintained within a narrow range through a complex interplay between intestinal absorption, exchange with intracellular and bone storage pools, and renal tubular reabsorption. The crucial regulated step in Pi homeostasis is the transport of Pi across the renal proximal tubule. Type II sodium-dependent phosphate (Na/Pi) cotransporter (NPT2) is the major molecule in the renal proximal tubule and is regulated by Pi, parathyroid hormone and by 1,25-dihydroxyvitamin D. Recent studies of inherited and acquired hypophosphatemia [X-linked hypophosphatemic rickets/osteomalacia (XLH), autosomal dominant hypophosphatemic rickets/osteomalacia (ADHR) and tumor-induced rickets/osteomalacia (TIO)], which exhibit similar biochemical and clinical features, have led to the identification of novel genes, PHEX and FGF23, that play a role in the regulation of Pi homeostasis. The PHEX gene, which is mutated in XLH, encodes an endopeptidase, predominantly expressed in bone and teeth, but not in kidney. FGF-23 may be a substrate of this endopeptidase and may therefore accumulate in patients with XLH. In the case of ADHR mutations in the furin cleavage site, which prevent the processing of FGF-23 into fragments, lead to the accumulation of a "stable" circulating form of the peptide which also inhibits renal Pi reabsorption. In the case of TIO, ectopic overproduction of FGF-23 overwhelms its processing and degradation by PHEX, leading to the accumulation of FGF-23 in the circulation and inhibition of renal Pi reabsorption. Mice homozygous for severely hypomorphic alleles of the Klotho gene exhibit a syndrome resembling human aging, including atherosclerosis, osteoporosis, emphysema, and infertility. The KLOTHO locus is associated with human survival, defined as postnatal life expectancy, and longevity, defined as life expectancy after 75. In considering the relationship of klotho expression to the dietary Pi level, the klotho protein seemed to be negatively controlled by dietary Pi.


Haplotype Analysis of DXS548 and FRAXAC1 Microsatellite Loci in Iranian Patients with Fragile X Syndrome.
Fragile X syndrome (FXS) is the most common cause of inherited mental retardation caused by expansion of a (CGG) repeat region up to 1000 repeat in 5' region of the FMR1 gene located in FRAXA locus Xq27.3. To better understand the mechanism involved in expansion of CGG region, the molecular characteristic of the flanking microsatellite markers in the region must be clarify in different populations. We aimed to examine the potential association between specific haplotype and the expanded AC-repeat region in cases and controls chromosomes. Forty unrelated FXS males and 62 unrelated normal males originating from various regions of Iran were haplotyped by analyzing two CA-repeat markers, FRAXAC1 and DXS548. Significant linkage disequilibrium was obtained between DXS548 and FRAXAC1 specific marker alleles and CGG repeat expansion among 40 fragile X cases compared to 62 normal controls. The frequencies of DXS548 and FRAXAC1 longer alleles in patients were significantly higher than that in control group. Two FRAXAC1 long alleles were only observed in cases, possibly due to concatenated mutations. The increase of heterozygosities in fragile X cases (DXS548 78.6%, FRAXAC1 64.6%) in comparison to the controls (DXS548 63.0%, FRAXAC1 47.0%) showed a multimodal distribution of fragile X associated alleles. Haplotype analyses with DXS548 and FRAXAC1 markers represented that haplotype distribution in the normal controls and FXS patients were significantly different, representing a weak founder effect.


Multicomponent gas chromatographic analysis of urinary steroids excreted by an infant with a defect in aldosterone biosynthesis.
The urinary steroids excreted by an infant with a salt-wasting syndrome due to a suspected defect in the 18-oxidation of corticosterone have been analysed by gas chromatography-mass spectrometry. The excretion of tetrahydroaldosterone was low (3.5 mug/24 h) whilst the excretion of 3alpha,11beta,21-trihydroxy-5alpha-pregnan-20-one (allo-tetrahydrocorticosterone) and other corticosterone metabolites was high (total about 2 mg/24 h). The excretion of cortisol metabolites was apparently normal for age (total about 2 mg/24 h) but 3alpha,11beta,17alpha,21-tetrahydroxy-5alpha-pregnan-20-one (allo-tetrahydrocortisol) rather than tetrahydrocortisone, was the major component of the group. The excretion of an 18-hydroxycorticosterone metabolite 3alpha,18,21-trihydroxy-5beta-pregnane-11,20-dione (18-hydroxytetrahydroCompound A) was higher than normal for infants of this age (between 50 and 200 mug/24 h), suggesting that the defect was in 18-hydroxysteroid dehydrogenase rather than 1,-hydroxylase. In addition, 18-hydroxytetrahydrocorticosterone, another metabolite of 18-hydroxycorticosterone was tentatively identified and it was found that the rate of excretion of this compound was of similar magnitude to 18-hydroxytetrahydroCompound A. The salt balance of the infant has been sucessfully controlled by salt administration (77 mEq./24 h) and treatment with Fludrocortisone (0.5 mg/day).


Foscarnet-Type Inorganic-Organic Hybrid Nanoparticles for Effective Antiviral Therapy.
[ZrO]2+[(FCN)0.4(OH)0.8]2- and Gd3+[FCN]3- inorganic-organic hybrid nanoparticles (IOH-NPs) are novel saline antiviral nanocarriers with foscarnet (FCN) as a drug anion. FCN as a pyrophosphate analogue serves as a prototype of a viral DNA polymerase inhibitor. FCN is used for the treatment of herpesvirus infections, including the drug-resistant cytomegalovirus (CMV) and herpes simplex viruses, HSV-1 and HSV-2. The novel [ZrO]2+[(FCN)0.4(OH)0.8]2- and Gd3+[FCN]3- IOH-NPs are characterized by aqueous synthesis, small size (20-30 nm), low material complexity, high biocompatibility, and high drug load (up to 44 wt % FCN per nanoparticle). The antiviral activity of the FCN-type IOH-NPs is probed for the human cytomegalovirus (HCMV). Moreover, the uptake of FCN-type IOH-NPs into vesicles, cytoplasm, and nuclei of nonphagocytic lung epithelial cells is evaluated. As a result, a promising antiviral activity of the FCN-type IOH-NPs that significantly outperforms freely dissolved FCN at the level of clinical formulations is observed, encouraging a future use of FCN-type IOH-NPs for the delivery of antivirals against respiratory viruses.


[Provisions for potentially raising the age limit for oocyte donation].
The introduction ofoocyte donation has provided a new perspective on reproduction for women with premature ovarian failure. In contrast to other forms ofassisted reproductive technology, the success ofoocyte donation is not affected by the age of the future mother. This has prompted a discussion on raising the age limit to allow older women to become pregnant. The health risk for the future mother is increased in older women compared with younger women but does not appear to be a sufficient reason to disqualify older women from undergoing the procedure. Lack of information regarding the effect of very late parenthood on the medical, psychological and social well-being of the future child poses the greatest concern. Should the age limit be extended for oocyte donation, treatment should be restricted to a research setting with a protocol for single-embryo transfer and extensive follow-up regarding the well-being of the future child.


CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
The CRISPR-Cas (Clustered Regularly Interspaced Short Palindrome Repeats--CRISPR associated proteins) system provides adaptive immunity in archaea and bacteria. A hallmark of CRISPR-Cas is the involvement of short crRNAs that guide associated proteins in the destruction of invading DNA or RNA. We present three fundamentally distinct processing pathways in the cyanobacterium Synechocystis sp. PCC6803 for a subtype I-D (CRISPR1), and two type III systems (CRISPR2 and CRISPR3), which are located together on the plasmid pSYSA. Using high-throughput transcriptome analyses and assays of transcript accumulation we found all CRISPR loci to be highly expressed, but the individual crRNAs had profoundly varying abundances despite single transcription start sites for each array. In a computational analysis, CRISPR3 spacers with stable secondary structures displayed a greater ratio of degradation products. These structures might interfere with the loading of the crRNAs into RNP complexes, explaining the varying abundancies. The maturation of CRISPR1 and CRISPR2 transcripts depends on at least two different Cas6 proteins. Mutation of gene sll7090, encoding a Cmr2 protein led to the disappearance of all CRISPR3-derived crRNAs, providing in vivo evidence for a function of Cmr2 in the maturation, regulation of expression, Cmr complex formation or stabilization of CRISPR3 transcripts. Finally, we optimized CRISPR repeat structure prediction and the results indicate that the spacer context can influence individual repeat structures.


Synchrotron-Based Infrared Microanalysis of Biological Redox Processes under Electrochemical Control.
We describe a method for addressing redox enzymes adsorbed on a carbon electrode using synchrotron infrared microspectroscopy combined with protein film electrochemistry. Redox enzymes have high turnover frequencies, typically 10-1000 s(-1), and therefore, fast experimental triggers are needed in order to study subturnover kinetics and identify the involvement of transient species important to their catalytic mechanism. In an electrochemical experiment, this equates to the use of microelectrodes to lower the electrochemical cell constant and enable changes in potential to be applied very rapidly. We use a biological cofactor, flavin mononucleotide, to demonstrate the power of synchrotron infrared microspectroscopy relative to conventional infrared methods and show that vibrational spectra with good signal-to-noise ratios can be collected for adsorbed species with low surface coverages on microelectrodes with a geometric area of 25 × 25 μm(2). We then demonstrate the applicability of synchrotron infrared microspectroscopy to adsorbed proteins by reporting potential-induced changes in the flavin mononucleotide active site of a flavoenzyme. The method we describe will allow time-resolved spectroscopic studies of chemical and structural changes at redox sites within a variety of proteins under precise electrochemical control.


BSA-AuNPs@Tb-AMP metal-organic frameworks for ratiometric fluorescence detection of DPA and Hg2.
An easy and effective strategy for synthesizing a ratiometric fluorescent nanosensor has been demonstrated in this work. Novel fluorescent BSA-AuNPs@Tb-AMP (BSA, bovine serum albumin; AMP, adenosine 5'-monophosphate; AuNPs, Au nanoparticles) metal-organic framework (MOF) nanostructures were synthesized by encapsulating BSA-AuNPs into Tb-AMP MOFs for the detection of 2,6-pyridinedicarboxylic acid (DPA) and Hg2+ . DPA could strongly co-ordinate with Tb3+ to replace water molecules from the Tb3+ center and accordingly enhanced the fluorescence of Tb-AMP MOFs. The fluorescence of BSA-AuNPs at 405 nm remained constant. While the fluorescence of BSA-AuNPs at 635 nm was quenched after Hg2+ was added, the fluorescence of Tb-AMP MOFs remained constant. Accordingly, a ratiometric fluorescence nanosensor was constructed for detection of DPA and Hg2+ . The ratiometric nanosensor exhibited good selectivity to DPA over other substances. The F545 /F405 linearly increased with increase of DPA concentration in the range of 50 nM to 10 μM with a detection limit as low as 17.4 nM. F635 /F405 increased linearly with increase of Hg2+ concentration ranging from 50 nM to 1 μM with a detection limit as low as 20.9 nM. Additionally, the nanosensor could be successfully applied for the determination of DPA and Hg2+ in running water.