A sequence-specific single-strand DNA binding protein that contacts repressor sequences in the human GM-CSF promoter.
NF-GMb is a nuclear factor that binds to the proximal promoter of the human granulocyte-macrophage colony stimulating factor (GM-CSF) gene. NF-GMb has a subunit molecular weight of 22 kDa, is constitutively expressed in embryonic fibroblasts and binds to sequences within the adjacent CK-1 and CK-2 elements (CK-1/CK-2 region), located at approximately -100 in the GM-CSF gene promoter. These elements are conserved in haemopoietic growth factor (HGF) genes. NF-GMb binding requires the presence of repeated 5'CAGG3' sequences that overlap the binding sites for positive activators. Surprisingly, NF-GMb was found to bind solely to single-strand DNA, namely the non-coding strand of the GM-CSF CK-1/CK-2 region. NF-GMb may belong to a family of single-strand DNA binding (ssdb) proteins that have 5'CAGG3' sequences within their binding sites. Functional analysis of the proximal GM-CSF promoter revealed that sequences in the -114 to -79 region of the promoter containing the NF-GMb binding sites had no intrinsic activity in fibroblasts but could, however, repress tumour necrosis factor-alpha (TNF-alpha) inducible expression directed by downstream promoter sequences (-65 to -31). Subsequent mutation analysis showed that sequences involved in repression correlated with those required for NF-GMb binding.


Expression of thioredoxin in patients with Graves' disease.
Thioredoxin (TRX), which is a stress-inducible protein with redox-active disulfide structures, has various biological activities by regulating DNA binding of transcription factors in cells. In Graves' disease that is among the common diseases of the thyroid, endogenous stresses that are induced by excess thyroid hormones or antibodies against thyroid stimulating hormone (TSH) receptors are responsible for the pathogenesis. The objective of this study was to examine the expression of TRX and to determine whether TRX is responsible for the pathogenesis of Graves' disease. The thyroid follicular cells were shown to express both TRX and vascular cell growth factors (VEGF) in all of the patients with Graves' disease by immunohistochemistry. In contrast, the expression of TRX or VEGF was not found in any of the normal thyroids. Serum levels of TRX were significantly elevated in patients with Graves' disease regardless of their thyroid function compared to those in healthy donors (122+/-16 versus 37+/-5 ng/ml, p<0.0001). Consecutive administration of iodine resulted in not only a reduction in serum levels of free triiodothyronine (T3) but also an increase in serum levels of TRX in the patients. These findings suggest that release of intracellular TRX from thyroid follicular cells in response to iodine resulted in suppression of T3 production. Taken together, TRX is highly produced under stress in Graves' disease and involved in regulating production of thyroid hormones. The investigation of biological behavior of this molecule may greatly help to understand pathogenesis of Graves' disease functions.


The relationship between the capillary structure and hemorrhage in gliomas.
A 48-year-old man was admitted with the sudden onset of symptoms of stroke caused by hemorrhage in an oligodendroglioma. Despite surgery and antiedema treatment, the patient died. Histological evaluation revealed an oligodendroglioma with calcified capillaries of the retiform type. To further investigate this phenomenon, a total of 160 gliomas were reviewed: 90 glioblastomas multiforme, 30 oligodendrogliomas, and 40 astrocytomas. Sufficient data were available for clinical evaluation in 100 cases. Of these, 5% (two oligodendrogliomas and three glioblastomas multiforme) were related to clinically significant hemorrhages. Of the remaining cases, microhemorrhages were found in 53.0% of the glioblastomas, in 56.7% of the oligodendrogliomas, and in 10.0% of the astrocytomas. In each case reviewed, the capillaries were assigned to one of three groups: axial, retiform, or glomeruloid. Statistical analysis revealed a significant association between hemorrhages and retiform capillaries in all three types of tumors, except that in oligodendrogliomas the statistical significance held true when calcification of the capillaries was also present. Glomeruloid-type capillaries were only weakly associated with hemorrhages, and no association was found for axial capillaries. A large-scale prospective study is necessary to more precisely assess the role of each of the three types of capillaries in hemorrhages of gliomas. Based on data available so far, patients with glial tumors with retiform capillaries, confirmed on biopsy, should be carefully monitored to exclude possible intratumoral hemorrhage.


An innovative technique of vertebral body stapling for the treatment of patients with adolescent idiopathic scoliosis: a feasibility, safety, and utility study.
Retrospective review. To report the feasibility, safety, and utility of vertebral body stapling without fusion as an alternative treatment for adolescent idiopathic scoliosis. The success rate of brace treatment of adolescent idiopathic scoliosis ranges from 50% to 82%. However, poor self-image and brace compliance are issues for the patient. An alternative method of treatment such as a motion-preserving vertebral body stapling to provide curve stability would be desirable. We retrospectively reviewed 21 patients (27 curves) with adolescent idiopathic scoliosis treated with vertebral body stapling. Patients were immature as defined by Risser sign <or=2. The concept of vertebral body stapling of the convex side of a patient with adolescent idiopathic scoliosis is feasible. The procedure was safe, with no major complications and three minor complications. One patient had an intraoperative segmental vein bleed resulting in an increased estimated blood loss of 1500 cc as compared to the average estimated blood loss of 247 cc for all patients. One patient had a chylothorax and one pancreatitis. No patient has had a staple dislodge or move during the follow-up period (mean 11 months, range 3-36 months), and no adverse effects specifically from the staples have been identified. Utility (defined as curve stability) was evaluated in 10 patients with stapling with greater than 1-year follow-up (mean 22.6 months) and preoperative curve <50 degrees. Progression of >or=6 degrees or beyond 50 degrees was considered a failure of treatment. Of these 10 patients, 6 (60%) remained stable or improved and 4 (40%) progressed. One of 10 (10%) in the stapling group had progressed beyond 50 degrees and went on to fusion. Six patients required stapling of a second curve, three as part of the primary surgery, and three as a second stage, because a second untreated curve progressed. The results need to be considered with caution, as the follow-up is still short. The data demonstrate that vertebral body stapling for the treatment of scoliosis in the adolescent was feasible and safe in this group of 21 patients. In the short-term, stapling appears to have utility in stabilizing curves of progressive adolescent idiopathic scoliosis.


A new sesquiterpene lactone from yacon leaves.
The chemical constituents of 60% EtOH extract of yacon leaves were separated to yield a new compound, together with four known compounds, which were isolated for the first time from yacon. The new compound was characterised and named as chlorodalin (1) on the basis of NMR (1D and 2D), HR-MS and other spectral methods. The cytotoxic activities of 1-5 were evaluated on two human tumour cell lines and the new compound showed significant cytotoxic activity.


An update on medium- and low-abundant blood plasma proteome of horse.
The main objectives of the study were to: (1) deeply analyse the serum protein composition of Equus caballus, (2) assess the effectiveness of the high-abundant protein depletion and improve the concentration of medium- and low-abundant proteins. The analysis were performed on the blood plasma of three healthy part-Arabian mares. The implementation of two-dimensional electrophoresis and matrix-assisted laser desorption/ionisation - time of flight mass spectrometry allowed us to establish a horse plasma proteome map. Serum proteins were resolved at pH 4 to 7, followed by 12% SDS-PAGE. As a result 136 spots were successfully identified, representing the products of 46 unique genes. Of these, 22 gene products have not been previously identified in horse serum/plasma samples using proteomic tools. Gene ontology analysis showed that almost 30% of all identified gene products belong to the coagulation and complement cascades. These results can undoubtedly serve as a useful and prospective prerequisite for the future analysis of horse plasma proteome changes in different physiological and pathophysiological conditions. The use of the medium- and low-abundant protein enrichment tool increased their abundance and allowed us to identify a higher number of protein gene products. The highest depletion efficiency was observed for the most abundant plasma proteins, that is albumin, IgG heavy chains and serotransferrin.


Diffuse axonal injury in head trauma.
Diffuse axonal injury (DAI) is one of the most common and important pathologic features of traumatic brain injury (TBI). The susceptibility of axons to mechanical injury appears to be due to both their viscoelastic properties and their high organization in white matter tracts. Although axons are supple under normal conditions, they become brittle when exposed to rapid deformations associated with brain trauma. Accordingly, rapid stretch of axons can damage the axonal cytoskeleton resulting in a loss of elasticity and impairment of axoplasmic transport. Subsequent swelling of the axon occurs in discrete bulb formations or in elongated varicosities that accumulate transported proteins. Calcium entry into damaged axons is thought to initiate further damage by the activation of proteases. Ultimately, swollen axons may become disconnected and contribute to additional neuropathologic changes in brain tissue. DAI may largely account for the clinical manifestations of brain trauma. However, DAI is extremely difficult to detect noninvasively and is poorly defined as clinical syndrome. Future advancements in the diagnosis and treatment of DAI will be dependent on our collective understanding of injury biomechanics, temporal axonal pathophysiology, and its role in patient outcome.


Suffodit inguina.
When Julius Caesar was stabbed, 23 times, on the Ides of March, at least one of the daggers is supposed to have gone into his groin. He wasn't the last Roman to have his privates attacked. And he wasn't the last primate. In competition for sexual access, gonads are occasionally targeted: canine incisions in monkey and ape scrota are not uncommon; and rumors had a number of Roman emperors--from Caligula and Nero, to Galba, Vitellius, Domitian, Commodus, Caracalla, Elagabalus, to Balbinus, Pupienus and Valerian over the course of the third century crisis--done in with their genitals at risk, or with their genitals cut off.


The hepatocarcinoma cell line HepG2 does not express a GHS-R1a-type ghrelin receptor.
The interaction of ghrelin, a 28-residue acylated peptide, with the growth hormone secretagogue receptor 1a (GHS-R1a) has been studied mostly in cells expressing a recombinant GHS-R1a. As awareness is growing on the importance to study G protein-coupled receptors in a natural environment, we studied the effect of ghrelin in the human hepatocellular HepG2 cell line because it has been described in literature to respond to ghrelin. Despite extensive efforts, we were not able to confirm mRNA expression of GHS-R1a by reverse transcription PCR, radioligand binding, or ghrelin-induced GHS-R1a receptor activation; therefore, we conclude that HepG2 cells do not express GHS-R1a. On the other hand, we confirmed a modest effect of ghrelin on the up-regulation of IRS-1 phosphorylation, which might suggest the existence of an alternative ghrelin receptor in HepG2 cells.


Nanobody-based immunosensing methods for safeguarding public health.
Immunosensing methods are biosensing techniques based on specific recognition of an antigen-antibody immunocomplex, which have become commonly used in safeguarding public health. Taking advantage of antibody-related biotechnological advances, the utilization of an antigen-binding fragment of a heavy-chain-only antibody termed as 'nanobody' holds significant biomedical potential. Compared with the conventional full-length antibody, a single-domain nanobody retaining cognate antigen specificity possesses remarkable physicochemical stability and structural adaptability, which enables a flexible and efficient molecular design of the immunosensing strategy. This minireview aims to summarize the recent progress in immunosensing methods using nanobody targeting tumor markers, environmental pollutants, and foodborne microbes.


[Solitary primary cerebral echinococcosis. Report of a case].
Echinococcosis is a widespread parasitic disease caused by Echinococcus granulosus. Hydatid cysts are mainly diagnosed in adults except for primary cerebral localisation which is electively observed in childhood owing to the early manifestation of signs and/or symptoms of the space-occupying mass. In July 1995 P.N., a 55-year-old woman, was referred to our attention. She complained of intense asthenia, cephalea not responding to NSAIDs and paroxysms of tremor. The anamnesis revealed close relationships with dogs since infancy and an attack of pleurisy of unknown etiology. The objective examination was negative except for an increased volume of the right-hand thyroid lobe. Hematochemical tests showed relative eosinophilia, a significant positivity of anti-thyroglobulin antibodies and antiperoxidase with normal thyroid function indices. Thyroid scan showed a multinodular goitre. Confirmation of eosinophilia suggested the performance of a parasitological examination of feces with negative results. ECG and EEG were normal. Persistent cephalea led to the performance of an encephalic CAT which revealed a cystic formation in the rolandic region, subsequently confirmed by encephalic MNR. The positivity of the Ghedini-Weinberg test led to the diagnosis of cerebral echinococcosis. Chest X-ray and hepatic scan excluded hydatid localisation in these organs. The case was diagnosed as solitary primary cerebral echinococcosis. Medical follow-up was commenced with albendazole for six months, after which a control encephalic CAT showed the unchanged size of the cysts. The patient consequently underwent surgical exeresis.


An antioxidant α-glucan from Cladina rangiferina (L.) Nyl. and its protective effect on alveolar epithelial cells from Pb2+-induced oxidative damage.
Air pollution is a serious global health problem nowadays. So, it is an emergency to pay sufficient attention to treat and prevent the diseases caused by air pollution, especially respiratory disease and lung damage. Cladina rangiferina (L.) Nyl. is an edible lichen that has been used in medicinal diets to treat respiratory and other diseases for over 500 years. In this study, a water-soluble polysaccharide, CRWP-P, was obtained from C. rangiferina by hot-water extraction, freeze-thawing separation, and Fehling reagent purification. Structural analysis showed that CRWP-P is a linear α-(1 → 3),(1 → 4)-d-glucan without branches. Its Mw was determined to be 1.05 × 105 Da. Its (1,3)-α-d-glucopyranosyl: (1,4)-α-d-glucopyranosyl ratio is approximately 1:2. Antioxidant activity assay showed that C. rangiferina polysaccharides, especially CRWP-P, had appreciable DPPH radical-scavenging activity and reducing power. Notably, they could effectively decrease cell breakdown and ROS generation, inhibit lipid peroxidation, increase key antioxidase activity, and promote glutathione redox cycling in Pb2+-oxidative injured A549 alveolar epithelium cells. Overall, the results of this study indicated that C. rangiferina polysaccharides, especially CRWP-P, have the potential to be natural antioxidants for the treatment of lung oxidative damage induced by lead of air pollutants.


Novel Fluorinated Porphyrins Sensitize Tumor Cells to Photodamage in Normoxia and Hypoxia: Synthesis and Biocompatible Formulations.
Hypoxia renders tumor cells refractory to treatment. One way to overcome this problem is the design of drug delivery systems that contain the antitumor agent within an oxygen supply medium. to evaluate whether the perfluorocarbon liquids (capable of retaining up to 50% v/v amounts of O2 gas) can be tools for delivery of photosensitizers to hypoxic tumors. We synthesized a series of compounds in which fluoroaliphatic or fluoroaromatic moieties were conjugated to the porphyrin ring in meso-positions. Two derivatives were tested as the solutions prepared either from a dimethylformamide stock ('free' formulation) or from a perfluorocarbon emulsion in which the photosensitizer is entrapped in the oxygenated medium. In the emulsion the hydrophobic photosensitizer and the gas transporting liquid represented a biocompatible composition. Free formulations or perfluorocarbon emulsions of fluorinated porphyrins evoked little-to-null dark cytotoxicity. In contrast, each formulation triggered cell death upon light activation. Photodamage in the presence of fluorinated porphyrins was achievable not only at normoxic (20.9% O2 v/v) conditions but also in hypoxia (0.5% O2). With new compounds dissolved in the medium the cell photodamage in hypoxia was negligible whereas a significant photodamage was achieved with the emulsions of fluorinated porphyrins. The derivative with the fluoroalkyl substituent was more potent than its structurally close analog carrying the fluoroaryl moiety. Our new fluorinated porphyrin derivatives, especially their emulsions in which the photosensitizer and the oxygenated medium are coupled into one phase, can be perspective for photoelimination of hypoxic tumor cells.


On-chip inter-modal Brillouin scattering.
Brillouin nonlinearities-which result from coupling between photons and acoustic phonons-are exceedingly weak in conventional nanophotonic silicon waveguides. Only recently have Brillouin interactions been transformed into the strongest and most tailorable nonlinear interactions in silicon using a new class of optomechanical waveguides that control both light and sound. In this paper, we use a multi-mode optomechanical waveguide to create stimulated Brillouin scattering between light-fields guided in distinct spatial modes of an integrated waveguide for the first time. This interaction, termed stimulated inter-modal Brillouin scattering, decouples Stokes and anti-Stokes processes to enable single-sideband amplification and dynamics that permit near-unity power conversion. Using integrated mode multiplexers to address separate optical modes, we show that circulators and narrowband filters are not necessary to separate pump and signal waves. We also demonstrate net optical amplification and Brillouin energy transfer as the basis for flexible on-chip light sources, amplifiers, nonreciprocal devices and signal-processing technologies.


Long non-coding RNA H19-mediated mouse cleft palate induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin.
Long non-coding RNAs (lncRNAs) are a novel class of transcripts, which are pervasively transcribed in the genome and a have greatly unknown biological function. Previous studies have identified that lncRNAs serve an important role in embryonic development. However, the function and mechanism of lncRNAs in the development of palate remains unclear. The aim of the present study was to investigate the role of lncRNA H19 in cleft palate (CP) development in mice. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a well-known teratogen that can induce CP. After establishing a CP mouse model by oral administration of TCDD in vivo, no significant differences were detected in the tail length and body weight of fetuses between the TCDD-treated and control groups during the embryonic days 12 to 17. Furthermore, the expression levels of lncRNA H19 and target gene insulin-like growth factor 2 (IGF2) presented specific embryo age-associated differences during the entire development of CP in mice. An inverse correlation was identified between lncRNA H19 and IGF2 expression levels in the CP model. In conclusion, these findings revealed that lncRNA H19 mediated the CP induced by TCDD in mice.


Enzyme-Linked Immunosorbent Spot Assay for the Detection of Wilms' Tumor 1-Specific T Cells Induced by Dendritic Cell Vaccination.
Despite recent advances in cancer immunotherapy and the development of various assays for T cell assessment, a lack of universal standards within immune monitoring remains. The objective of this study was to evaluate the enzyme-linked immunosorbent spot (ELISpot) assay in comparison with major histocompatibility complex-tetramer analysis in the context of dendritic cell (DC)-based cancer immunotherapy. The ELISpot assay was performed on peripheral blood mononuclear cells to assess reproducibility, daily precision, and linearity using HLA-A*24:02-restricted Cytomegalovirus peptide. Wilms' tumor 1 (WT1) antigen-specific cytotoxic T cells were then evaluated by both the ELISpot assay and WT1 tetramer analysis in peripheral blood from 46 cancer patients who received DC vaccinations pulsed with human leukocyte antigen (HLA)-A*24:02-restricted modified WT1 peptides. The ELISpot assay was proven to have reproducibility (coefficient of variation (CV) ranged from 7.4% to 16.3%), daily precision (CV ranged from 5.0% to 17.3%), and linearity (r = 0.96-0.98). WT1-specific immune responses were detected by the ELISpot assay in 34 out of 46 patients (73.9%) post-vaccination. A Spearman's rank-correlation coefficient of 0.82 between the ELISpot assay and WT1 tetramer analysis was obtained. This is the first report of a comparison of an ELISpot assay and tetramer analysis in the context of dendritic cell (DC)-based cancer immunotherapy. The ELISpot assay has reproducibility, linearity, and excellent correlation with the WT1 tetramer analysis. These findings suggest that the validated ELISpot assay is useful to monitor the acquired immunity by DC vaccination targeting WT1.


Long-term evaluation of vertebral artery injuries following cervical spine trauma using magnetic resonance angiography.
A prospective study to determine the long-term outcome of traumatically induced vertebral artery injuries. Magnetic resonance angiography was performed at the time of cervical injury and at a follow-up office visit. To determine the long-term outcome in terms of arterial flow competency of traumatically induced vertebral artery injuries. Vertebral artery injury associated with cervical spine trauma has been well documented; however its healing or nonhealing potential has not been elucidated. During the 7-month period from July 1993 to January 1994, all patients admitted to the authors' institution with cervical spine injuries underwent magnetic resonance imaging and magnetic resonance angiography of the cervical spine to determine the patency of their vertebral arteries. Magnetic resonance angiography was performed at the time of injury and at a follow-up office visit. Twelve of 61 patients were found to have a lack of signal flow within one of their vertebral vessels during this study period. Eighty-three percent of the patients (five of six) who were available for follow-up observation in this study did not manifest flow reconstitution of their vertebral arteries after an average 25.8-month follow-up period. According to these data, most patients with vertebral artery injuries after nonpenetrating cervical spine trauma do not reconstitute flow in the injured vertebral arteries. This lack of flow must be considered if future surgery in this region of the cervical spine is contemplated.


Effects of glucose and disorders in lipid metabolism on cytokine levels and cognitive impairment in the olanzapine-induced obesity rat model.
The aim of the study was to explore the effects of increased levels of blood sugar and cytokines on impaired cognitive function in the olanzapine-induced obesity rat model. A total of 40 rats were randomly divided into 2 groups; the control and olanzapine groups (N = 20 per group). The control rats were fed regular food, while the olanzapine rats received olanzapine-enriched (1.2 mg/kg) food by gavage for 4 weeks to establish the olanzapine-induced obese rat model. Enzyme-linked immunosorbent assays were used to measure the serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and C-reactive protein (CRP). Serum glucose content was measured by biochemical colorimetry. Learning and memory capacity was measured using a Y-maze, and the time before escape from a Morris water maze was recorded. Body weight and levels of blood glucose, lipids, TNF-α, IL-6, and CRP increased in the olanzapine group. In addition, the number of shocks received before reaching the learning and memory standard and the time before escape from the Morris water maze were higher in the olanzapine group than in the control group. Olanzapine causes disorders in glucose and lipid metabolism. Increase in blood glucose promotes the toxicity of cytokines and leads to cognitive dysfunction in rats.


Seeds of HOPE: Incorporating Community-Based Strategies to Implement a Weight-Loss and Empowerment Intervention in Eastern North Carolina.
Seeds of HOPE (Health, Opportunities, Partnerships, and Empowerment) was a community-based participatory research project designed to implement an evidence-based weight loss and empowerment intervention aimed at increasing self-efficacy through peer support. HOPE Works, its model, has yielded significant weight loss and increases in participants' ability to set and achieve goals, including in consumption of fruits and vegetables and in physical activity.METHODS The Seeds of HOPE project was implemented through 3 peer leader recruitment approaches using Circle Leaders. Study participants met in hope circles to discuss various topics aimed at developing healthier lifestyles. Changes at 6-month intervals were assessed in health behaviors, weight, and hope. One-way ANOVA was used to test differences in mean change in weight and hope across the 3 recruitment approaches.RESULTS Study participants (N = 146) lost weight (-3.3 pounds, P = 0.003, 95% CI: -5.40 - -1.27). Fruit consumption significantly increased (P = 0.04, 95% CI: 0.01-0.41) while no changes in vegetable consumption or physical activity were observed. No differences based on different leader recruitment approaches were found.LIMITATIONS There were limitations in the power to fully detect differences across Circle Leader recruitment approaches; loss-to-follow up and representativeness of the sample were present. Further research is needed to determine if these approaches would be successful in other rural counties, with different genders, or with individuals in urban and/or higher socioeconomic populations.CONCLUSIONS Participants found modest success in weight loss and fruit consumption. Authors find that community engagement strategies are helpful in finding mutually acceptable adaptations to implement interventions such as Seeds of HOPE in diverse communities.


Glutamate, excitatory amino acid transporters, Xc- antiporter, glutamine synthetase, and gamma-glutamyltranspeptidase in human corneal epithelium.
The viability and functions of the corneal epithelium are dependent in large measure on the active uptake of nutrients, growth factors, and amino acids from stroma and tear. The present study presents the cellular distribution(s) of glutamate, the Na(+)-dependent glutamate/aspartate transporters (excitatory amino acid transporters; EAAT1-5), Na(+)-independent glutamate/cystine exchanger (Xc(-) antiporter) subunits (xCT light chain and 4F2hc heavy chain), glutamine synthetase (GS), and gamma-glutamyltranspeptidase (GGT) in human corneal epithelium. Glutamate, EAAT1-5, xCT/4F2hc, GS, and GGT immunoreactive proteins were detected by immunofluorescence microscopy. Human corneal GGT activity was quantified using a standard colorimetric assay. Glutamate, EAAT3>2>1, xCT/4F2hc, and GGT proteins were detected in the columnar and wing cells. Glutamate was reduced or absent in the EAAT negative, Xc(-) antiporter, and GS positive outer wing cell and flat superficial epithelial cell layers. All EAATs (EAAT3>4/5>1/2), xCT/4F2hc, GS, and GGT were detected in flat superficial epithelial cell layer. The localization of glutamate, multiple EAATs, Xc(-) antiporter proteins, and GGT to columnar and superficial epithelial cell layers suggests uptake of glutamate and cystine from the stroma and tear and supports their importance in regulation of glutamate/cystine and glutathione (GSH; a tripeptide of glutamate, cystine, and glycine) in the human cornea epithelium. In addition, the low glutamate levels in outer wing and flat superficial epithelial cells positive for Xc(-) antiporter and GS are consistent with exchange of glutamate by Xc(-) antiporter for extracellular cystine utilized in GSH synthesis and support coupling of ammonia detoxification with glutamate degradation by GS.


Rapid Detection of Diazepam Injection Based on Surface Enhanced Raman Spectroscopy.
Diazepam injection was detected based on a droplet surface enhanced Raman spectroscopy (SERS) platform, which overcomes the disadvantages of the poor uniformity and time-consuming sample treatment process of the conventional "drop and dry" detection strategy. The Raman peak positions of diazepam injection were determined and identified, they are mainly located in the 689, 1002, 1170 and 1598 cm-1, etc. Different concentrations of diazepam injection were detected. It was found that the intensity of 1002 cm-1 increases linearly with concentration in the range from 0.05 to 10 μg/mL and the linear correlation coefficient is 0.988. The limit of detection can reach 0.05 μg/mL. The SERS method is easy, fast and efficient. The results are accurate and reliable. It has favorable application potential in the detection of diazepam injection.


Transdermal estrogens in the changing landscape of hormone replacement therapy.
Views and clinical practice on hormone replacement therapy (HRT) for postmenopausal women have varied widely over the last several decades. The 1990s showed a dramatic decline in prescribing and only recently are prescription rates increasing again. Use of HRT is first line for women suffering from the symptoms of menopause. For a certain population of postmenopausal women, HRT is recommended for prevention of fracture. More consideration is being given to patient risk factors, side effects, and formulations available. Transdermal patch estrogen formulations have been on the market for many years and are becoming more common although many physicians are less familiar with the various options. Transdermal preparations are worth consideration with regard to less adverse effects as there is the avoidance of liver metabolism and accumulation of metabolites with antiestrogenic activity. Data suggests that transdermal patch estrogen preparations are both safer and potentially more efficacious than oral estrogen therapy. Guidelines recommend the addition of progesterone for women who have not had a hysterectomy. Transdermal patch estrogen has a role for women as a choice for individualized therapy and providers need to be familiar with the formulations as well as unique challenges and advantages.


SPONTANEOUS SUPRACHOROIDAL HEMORRHAGE ASSOCIATED WITH IATROGENIC COAGULOPATHY.
To present a case of spontaneous suprachoroidal hemorrhage in a patient on anticoagulation. Single case report. A 68-year-old woman with a history of hypertension, atherosclerosis, and heart failure presented with massive spontaneous suprachoroidal hemorrhage in the left eye. She had no known ophthalmic pathology. She was taking warfarin and she had a high international normalization ratio. Despite medical and surgical therapy, there was no improvement and vision deteriorated to no light perception. Spontaneous expulsive suprachoroidal hemorrhage is a rare entity. Advanced age, systemic anticoagulation, and hypertension are strong risk factors. It is important to monitor international normalization ratio in warfarinized patients; particularly, uncontrolled high levels may cause spontaneous suprachoroidal hemorrhage in the absence of predisposing ocular pathology.


Activity of the enzymes involved in the synthesis of exopolysaccharide precursors in an overproducing mutant ropy strain of Streptococcus thermophilus.
The activities of some enzymes belonging to the Leloir pathway, phosphoglucomutase, UDP-glucose pyrophosphorylase, UDP-galactose 4-epimerase and galactose 1-P uridyl transferase, were studied in a wild ropy, a non-ropy and an overproducing mutant ropy strain of Streptococcus thermophilus. These activities were assayed over successive culture transfers along with exocellular polysaccharide (EPS) production. The overproducing mutant ropy strain showed increments in polysaccharide production over successive culture transfers, as opposed to reductions in production by the wild ropy strain. The observed variations among strains in the enzyme activities that were analysed in relation to EPS production suggest their involvement in the synthesis of sugar-nucleotide EPS precursors.


HLA DR and DQ distribution in normal human ocular structures.
HLA DR and DQ distribution was investigated in normal human ocular tissues, together with class I antigens and immunocompetent cell subsets, by immunofluorescence and immunoperoxidase procedures. In the anterior segment, our findings, consistent with those of previous reports, showed the wide distribution of class I antigens, specially in the corneal epithelium, while class II antigens were restricted to very rare cells scattered in the conjunctiva, the peripheral cornea and the stroma of the ciliary processes. Some non pigmented epithelial cells of the ciliary processes were HLA DR and DQ positive. In the posterior segment, class I antigens were abundantly represented in the choroid and the retinal layers. Few HLA DR and DQ positive cells were seen in the choroid, similar to those found in the anterior segment. Normal RPE did not react with any monoclonal antibody, but numerous cells located in the retina were strongly HLA DR and DQ positive, all around the blood vessels, and not at the sites of endothelial cells. The characterization of those cells, which could be hypothetized as pericytes needs further studies but suggests close relationships between neuroretina and the immune system. This study may provide insight in the implication of the immune system in many poorly understood ocular diseases.


Insights into the oxidation and decomposition of CO on Au/alpha-Fe2O3 and on alpha-Fe2O3 by coupled TG-FTIR.
CO oxidation and decomposition behaviors over nanosized 3% Au/alpha-Fe2O3 catalyst and over the alpha-Fe2O3 support were studied in situ via thermogravimetry coupled to on-line FTIR spectroscopy (TG-FTIR), which was used to obtain temperature-programmed reduction (TPR) curves and evolved gas analysis. The catalyst was prepared by a sonication-assisted Au colloid based method and had a Au particle size in the range of 2-5 nm. Carburization studies of H 2-prereduced samples were also made in CO gas. According to gravimetry, for the 3% Au/alpha-Fe2O3 catalyst, there were three distinct stages of CO interaction with the Au catalyst but only two stages for the catalyst support. At low temperatures (<or=100 degrees C), only the Au catalyst had a rapid weight loss, which confirmed that CO reacted with highly active absorbed oxygen species and/or OH species which were associated with and promoted by the Au nanoparticles. Around 300 degrees C, both the catalyst and support samples experienced the reduction of Fe2O3 to Fe3O4, while above 400 degrees C further reduction to FeO and Fe metal took place. Au played no part in the kinetics of Fe3O4 formation because lattice O mobility was rate-limiting. At higher temperature where Fe3O4 was further reduced to FeO and Fe 0, the initially formed metallic Fe 0 nuclei could decompose CO molecules and release O species. Both this coproduced O species and the lattice oxygen could react with CO molecules. Thus, the CO oxidation was not limited by the mobility of lattice oxygen, and the catalytic function of Au was revealed again. Carburization of metallic Fe, created by prereduction in H 2, revealed a distinct weight gain at 350 degrees C corresponding to Fe 3C formation, as subsequently confirmed by X-ray diffraction (XRD). Sustained carbon deposition ensued at 450 degrees C. In the cases of the 3% Au/gamma-Al 2O 3 and Au/ZrO 2 catalysts prepared by the same method, however, after exposure to CO in the same temperature range, no carbon deposit was observed, indicating that although Au nanoparticles could activate the absorbed oxygen molecules at low temperatures, they were not able to activate the lattice oxygen in the three catalyst supports or to dissociate the CO molecules directly.


Follistatin attenuates radiation-induced fibrosis in a murine model.
Fibrosis can be a disabling, severe side effect of radiotherapy that can occur in patients, and for which there is currently no effective treatment. The activins, proteins which are members of the TGFβ superfamily, have a major role in stimulating the inflammatory response and subsequent fibrosis. Follistatin is an endogenous protein that binds the activins virtually irreversibly and inhibits their actions. These studies test if follistatin can attenuate the fibrotic response using a murine model of radiation-induced fibrosis. C57BL/6 mice were subcutaneously injected with follistatin 24 hours prior to irradiation. Mice were irradiated in a 10 x 10 mm square area of the right hind leg with 35 Gy and were given follistatin 24 hours before radiation and three times a week for six months following. Leg extension was measured, and tissue was collected for histological and molecular analysis to evaluate the progression of the radiation-induced fibrosis. Leg extension was improved in follistatin treated mice compared to vehicle treated mice at six months after irradiation. Also, epidermal thickness and cell nucleus area of keratinocytes were decreased by the follistatin treatment compared to the cells in irradiated skin of control mice. Finally, the gene expression of transforming growth factor β1 (Tgfb1), and smooth muscle actin (Acta2) were decreased in the irradiated skin and Acta2 and inhibin βA subunit (Inhba) were decreased in the irradiated muscle of the follistatin treated mice. Follistatin attenuated the radiation-induced fibrotic response in irradiated mice. These studies provide the data to support further investigation of the use of follistatin to reduce radiation-induced fibrosis in patients undergoing radiotherapy for cancer.


Auriculatone Sulfate Effectively Protects Mice Against Acetaminophen-Induced Liver Injury.
Acetaminophen (APAP) overdose is very common worldwide and has been widely recognized as the leading cause of drug-induced liver injury in the Western world. In our previous investigation, auriculatone, a natural product firstly obtained from Aster auriculatus, has demonstrated a potent protective effect against APAP-induced hepatotoxicity in HL-7702 cells. However, the poor water solubility and low bioavailability restrict its application. Auriculatone sulfate (AS) is a sulfated derivative of auriculatone with highly improved water-solubility. Hepatoprotective effects against APAP-induced liver injury (AILI) showed that intragastric pretreatment with AS at 50 mg/kg almost completely prevented mice against APAP-induced increases of serum alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and ATPase. Histological results showed that AS could protect the liver tissue damage. In addition, AS pretreatment not only significantly retained hepatic malondialdehyde and the activities of glutathione, superoxide dismutase, and glutathione peroxidase at normal levels, but also markedly suppressed the increase of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 levels in mouse liver caused by overdose APAP. Immunohistochemical analysis showed that AS obviously attenuated the expression of CD45 and HNE in liver tissue. Further mechanisms of action investigation showed that inhibition of cytochrome P450 3A11 (CYP 3A11) and CYP2E1 enzymatic activities (but not that of CYP1A2) was responsible for APAP bioactivation. In conclusion, AS showed a hepatoprotective effect against AILI through alleviating oxidative stress and inflammation and inhibiting CYP-mediated APAP bioactivation. It may be an effective hepatoprotective agent for AILI and other forms of human liver disease.


Hsp90 inhibitors identified from a library of novobiocin analogues.
Novobiocin is a C-terminal inhibitor of the Hsp90 protein folding machinery, which is responsible for the conformational maturation of numerous proteins involved in cancer growth and survival. Due to novobiocin's poor inhibitory activity ( approximately 700 muM), very little attention has been paid toward the development of novobiocin analogues for Hsp90 inhibition. In this study, a parallel library of 20 novobiocin derivatives was prepared and the biological activity of each evaluated by Western blot analysis of Hsp90 client proteins. A4 was found to be a potent inhibitor of Hsp90 as determined by its ability to cause the degradation of several Hsp90 client proteins in both breast and prostate cancer cell lines. In the presence of 1 muM A4, several Hsp90 client proteins were degraded, including AKT, Her2, Hif-1alpha, and the androgen receptor.


Telephone listening ability for hearing-impaired individuals.
Forty-five subjects with bilateral sensorineural hearing losses were divided into three groups (N = 15/group) based on the audiometric configuration of their test ear. The subjects were evaluated in three listening conditions by assessing their word discrimination ability by presenting stimuli at 86 dB SPL (simulating the output of a standard telephone handset ) and most comfortable loudness (MCL) (simulating the output of a telephone amplifier handset ). The three listening conditions were TDH-39 earphone, unaided telephone, and aided telephone with their personal hearing aid acoustically coupled to a telephone handset . Results indicated that subjects with precipitous hearing losses had similar word discrimination scores for all three listening conditions at 86 dB SPL and MCL. Subjects with gradual slope or flat hearing losses had similar word discrimination scores with a TDH-39 earphone, unaided telephone, and their hearing aids acoustically coupled to a telephone handset at 86 dB SPL. The subjects with gradual slope or flat losses also had similar word discrimination scores across the listening modes at MCL, but they had better word discrimination scores in all listening modes when the word lists were presented at MCL compared with 86 dB SPL.


Bottom-up Construction of a Primordial Carboxysome Mimic.
We have constructed a synthetic mimic of the carboxysome, a cyanobacterial carbon-fixing organelle. Using an electrostatic tagging system, we coencapsulated the two key carboxysomal enzymes, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and carbonic anhydrase (CA), in an engineered protein cage based on lumazine synthase. A statistically significant kinetic effect of coencapsulated CA on RuBisCO activity was not observed under ambient or oxygen saturated conditions, suggesting that enzyme proximity alone may not be the key determinant in carboxysome function. The capsid shell protected the enzyme from proteolytic damage, a factor that could have provided early cyanobacteria with an evolutionary benefit. Our strategy to coencapsulate different proteins can easily be extended to other sequentially acting enzymes and lays down principles for developing artificial organelles to control biosynthetic pathways in vivo.


An efficient algorithm for classical density functional theory in three dimensions: ionic solutions.
Classical density functional theory (DFT) of fluids is a valuable tool to analyze inhomogeneous fluids. However, few numerical solution algorithms for three-dimensional systems exist. Here we present an efficient numerical scheme for fluids of charged, hard spheres that uses O(N log N) operations and O(N) memory, where N is the number of grid points. This system-size scaling is significant because of the very large N required for three-dimensional systems. The algorithm uses fast Fourier transforms (FFTs) to evaluate the convolutions of the DFT Euler-Lagrange equations and Picard (iterative substitution) iteration with line search to solve the equations. The pros and cons of this FFT/Picard technique are compared to those of alternative solution methods that use real-space integration of the convolutions instead of FFTs and Newton iteration instead of Picard. For the hard-sphere DFT, we use fundamental measure theory. For the electrostatic DFT, we present two algorithms. One is for the "bulk-fluid" functional of Rosenfeld [Y. Rosenfeld, J. Chem. Phys. 98, 8126 (1993)] that uses O(N log N) operations. The other is for the "reference fluid density" (RFD) functional [D. Gillespie et al., J. Phys.: Condens. Matter 14, 12129 (2002)]. This functional is significantly more accurate than the bulk-fluid functional, but the RFD algorithm requires O(N(2)) operations.


A patient-calibrated individual wax-up as an essential tool for planning and creating a patient-oriented treatment concept for pathological tooth wear.
Tooth wear is generally a physiological process; however, it can also be a pathological condition associated with attrition, abrasion, and erosion processes in which the quantity of tooth loss is atypical for the patient's age. Tooth substance loss often causes functional problems, symptoms of discomfort/pain, and esthetic impairments. Patients presenting this condition frequently need restorative treatment, combined with an increase in the vertical dimension of occlusion (VDO). A diagnostic wax-up (DWU) provides assistance in determining a new occlusal plane and VDO, and is a fundamental tool considering the backward planning involved in the execution of a complex rehabilitation. In cases of severe tooth wear, preventive measures must be taken. Yet, when a restorative intervention is needed, the use of a DWU, in conjunction with a mock-up, helps to achieve a predictable and satisfactory outcome. This article presents a case report of a young female patient with pathological tooth wear, related symptomatology, and esthetic complaints. Initially, a DWU and a diagnostic mock-up were created. After esthetic and functional corrections, a second DWU was generated and transferred to the patient's mouth by means of a therapeutic mock-up. This mock-up was used as a trial restoration for the pretreatment phase, to evaluate the new VDO/esthetics/function and to guide tooth preparation. Lithium disilicate (LS2) occlusal onlays were used to stabilize the VDO, and anterior LS2 and feldspathic veneers, combined with direct composite restorations, were executed to reach the planned minimally invasive result.


Fragmented population structure of plasmodium falciparum in a region of declining endemicity.
The population genetic structure of Plasmodium falciparum differs between endemic regions, but the characteristics of a population recently fragmented by effective malaria control have been unknown. Genotypic analysis of 10 microsatellite loci widely separated in the parasite genome was conducted on 288 P. falciparum isolates from 8 foci in Malaysian Borneo, a region in which malaria incidence has been progressively reduced. Within all P. falciparum foci, moderate levels of allelic diversity were found, but levels of multilocus linkage disequilibrium were extremely variable. The population with the highest proportion of mixed-clone infections also had the highest allelic diversity and nonsignificant linkage disequilibrium. In contrast, several populations showed evidence of clonal expansion, and one offshore island population had exceptionally high levels of linkage disequilibrium. Genetic differentiation between many populations was very high and strongly associated with the geographical distance between them. High levels of differentiation and contrasting population structure among P. falciparum populations in Malaysian Borneo indicate that they are genetically independent. This supports the feasibility of individually eradicating the remaining P. falciparum foci.


Source identification of ambient aerosols over an urban region in western India.
A first-of-its-kind source apportionment study of the Ahmedabad, India aerosol was conducted in order to determine the major sources contributing to the measured total suspended particles (TSPs). TSP samples were collected approximately once every ten days between May 2000 and January 2003, and analyzed for TSP mass, anions, cations, and elemental concentrations. An advanced factor analysis technique, positive matrix factorization (PMF) was applied to the measured concentrations and six factors were resolved. The model resolved factors included airborne regional dust, calcium carbonate rich dust, biomass burning/vehicular emissions, secondary nitrate/sulfate, marine aerosol, and smelter. Among the resolved factors, airborne regional dust was the highest contributor to the measured TSP mass followed by calcium carbonate rich dust with their average contributions accounting for 57.9 and 19.0%, respectively. Thus, crustal factors were the most dominant sources of TSP in Ahmedabad accounting for nearly 77% of the mass. Potential source contribution function (PSCF) analysis identified parts of Madhya Pradesh and Uttar Pradesh, regions in southwestern Pakistan along the Indo-Gangetic Plain (IGP), and southern Iran as potential source locations for the airborne regional dust factor. In contrast, Rajasthan and Madhya Pradesh, and parts of northern Pakistan were identified as potential source locations for the calcium carbonate rich dust factor. It is hypothesized that aerosol contributions from several limestone quarries in Rajasthan and Madhya Pradesh may have resulted in this factor being calcium carbonate enriched.


Adenosquamous carcinoma of the pancreas.
Adenosquamous carcinoma of the pancreas is an unusual tumor. The authors review the case report of a 70-year-old male. Radiologic and computer tomography (CT) findings for adenosquamous carcinoma were indistinguishable from adenocarcinoma of the pancreas.


Hypocholesterolemic activity of cornelian cherry (Cornus mas L.) fruits.
Background Lipid profile disturbances are important risk factors for cardiovascular events in patients with diabetes mellitus and finding safe and multifaceted agents is persuaded in this regard. This study aimed to evaluate the effect of cornelian cherry dried powder (CCDP) on serum lipid profile as well as liver antioxidant capacity, HMG-CoA reductase level and activity, and LDL receptor level in streptozotocin-induced diabetic rats. Methods Forty-eight male adult Wistar rats were randomly allocated into eight equal groups and were treated for 4 weeks as follows: negative control (normal rats, basic diet); positive control (diabetic rats, basic diet), T1 to T4 groups: diabetic rats fed with basic diet containing 0.25, 0.5, 1 and 2 g/ 100 g BW CCDP, respectively; T5: diabetic rats fed with basic diet plus 10 mg/kg lovastatin in drinking water and T6: normal rats fed with basic diet containing 1 g/ 100 g BW CCDP. Results Administration of CCDP had no significant effect on serum glucose levels in diabetic rats however decreased total cholesterol, low-density lipoprotein cholesterol (LDL-C) and increased high-density lipoprotein cholesterol (HDL-C) levels and liver antioxidant capacity as compared to positive control rats (p<0.05). Although HMG-CoA reductase level showed a significant decrease only in T3 group, its activity was reduced in all diabetic CCDP and lovastatin-treated groups as compared to positive control. LDL receptor level remained statistically the same among positive control and CCDP-treated groups. Conclusions In conclusion, the present study confirms hypocholesterolemic effect of CCDP in diabetic rats and demonstrated that this effect was at least partly due to inhibition of liver HMG-CoA reductase activity.


Prognostic value of lymphovascular space invasion according to the molecular subgroups in endometrial cancer.
Lymphovascular space invasion (LVSI) is a known prognostic factor for oncological outcome in endometrial cancer patients. However, little is known about the prognostic value of LVSI among the different molecular subgroups. The aim of this study was to determine the prognostic dependence of LVSI from the molecular signature. This study included endometrial cancer patients who underwent primary surgical treatment between February 2004 and February 2016 at the Karolinska University Hospital, Sweden and the Bern University Hospital, Switzerland (KImBer cohort). All cases had complete molecular analysis performed on the primary tumor according to the WHO Classification of Tumors, 5th edition. LVSI was reviewed by reference pathologists for all pathology slides. A total of 589 endometrial cancer patients were included in this study, consisting of 40 POLEmut (polymerase epsilon ultramutated), 198 MMRd (mismatch repair deficient), 83 p53abn (p53 abnormal), and 268 NSMP (non-specific molecular profile) cases. Altogether, 17% of tumors showed LVSI: 25% of the POLEmut, 19% of the MMRd, 30% of the p53abn, and 10% of the NSMP cases. There was a significant correlation of LVSI with lymph node metastasis in the entire study cohort (p<0.001), remaining significant in the MMRd (p=0.020), p53abn (p<0.001), and NSMP (p<0.001) subgroups. Mean follow-up was 89 months (95% CI 86 to 93). The presence of LVSI significantly decreased recurrence-free survival among patients with MMRd, p53abn, and NSMP endometrial cancer, and overall survival in patients with p53abn and NSMP tumors. In patients with NSMP endometrial cancer, evidence of substantial LVSI remained a significant independent predictor of recurrence in multivariable Cox regression analysis including tumor stage and grade (HR 7.5, 95% CI 2.2 to 25.5, p=o.001). The presence of LVSI was associated with recurrence in each subgroup of patients with MMRd, p53abn, and NSMP endometrial cancer, and LVSI remained an independent predictor of recurrence in NSMP endometrial cancer patients.


Mitochondrial localization of superoxide dismutase is required for decreasing radiation-induced cellular damage.
We investigated the importance of mitochondrial localization of the SOD2 (MnSOD) transgene product for protection of 32D cl 3 hematopoietic cells from radiation-induced killing. Four plasmids containing (1) the native human copper/zinc superoxide dismutase (Cu/ZnSOD, SOD1) transgene, (2) the native SOD2 transgene, (3), the SOD2 transgene minus the mitochondrial localization leader sequence (MnSOD-ML), and (4) the SOD2 mitochondrial leader sequence attached to the active portion of the SOD1 transgene (ML-Cu/ZnSOD) were transfected into 32D cl 3 cells and subclonal lines selected by kanamycin resistance. Clonogenic in vitro radiation survival curves derived for each cell clone showed that Cu/ZnSOD- and MnSOD-ML-expressing clones had no increase in cellular radiation resistance (D0=0.89 +/- 0.01 and 1.08 +/- 0.02 Gy, respectively) compared to parent line 32D cl 3 (D0=1.15 +/- 0.11 Gy). In contrast, cell clones expressing either SOD2 or ML-Cu/ZnSOD were significantly radioresistant (D0=2.1 +/- 0.1 and 1.97 +/- 0.17 Gy, respectively). Mice injected intraesophageally with SOD2-plasmid/liposome (MnSOD-PL) complex demonstrated significantly less esophagitis after 35 Gy compared to control irradiated mice or mice injected intraesophageally with Cu/ZnSOD-PL or MnSOD-ML-PL. Mice injected with intraesophageal ML-Cu/ZnSOD-PL showed significant radioprotection in one experiment. The data demonstrate the importance of mitochondrial localization of SOD in the in vitro and in vivo protection of cells from radiation-induced cellular damage.


[Studies on enrichment broth for verotoxin-producing Escherichia coli (VTEC) O157].
Growth of verotoxin-producing Escherichia coli (VTEC) O157 in conventionally recommended pre-enrichment broth media at different temperatures was evaluated. In addition, sensitivity of VTEC O157 isolates to several antibacterial drugs, which were added to the selective enrichment broth, was tested. All five isolates of VTEC O157 tested grew well in trypticase soy broth (TSB) at 36 degrees C and 42 degrees C, while the growth of one isolate was markedly suppressed in TSB supplemented with cefixime (CFIX), potassium tellurite (PT), and vancomycin (TSB-CTV) even at 36 degrees C. A significant growth suppression was also observed in three of the isolates cultured in novobiocin (NB)-supplemented modified EC broth (mEC-NB) at 42 degrees C. In mEC-NB after 24-hr incubation at 36 degrees C, the five VTEC O157 isolates grew well, although one isolate was slightly suppressed during the first 8 hours. Minimum growth inhibitory concentrations of CFIX, NB and PT for a total of 90 clinical and environmental isolates of VTEC O157 were all above the concentrations usually prescribed for mEC-NB and TSB-CTV. These findings suggest that mEC-NB and TSB-CTV should be used at 36 degrees C for growth of VTEC O157 and that use of a nonselective pre-enrichment broth medium (i.e. TSB) together with a selective one (i.e. TSB-CTV or mEC-NB) is necessary for successful isolation of VTEC O157 from various specimens.


Language Use and Generation Status are Associated with Chronic Pain Differences in Mexican Americans.
Little is known about the pain experience of the Mexican American (MA) population. We investigated the associations between language use and generation status with chronic pain prevalence, health insurance coverage, and analgesic medication use. We examined 3373 MA respondents from the National Health and Nutrition Examination Survey. We found higher levels of English use and generation status were associated with higher odds of reporting chronic pain. For respondents reporting chronic pain, higher levels of English use and generation status were associated with higher odds of being covered by health insurance, lower odds of having a period of time last year without health insurance, and higher odds of being prescribed any analgesic medication, especially opioid medications. We found language use and generation status play a role in MAs' experience, access, and treatment of chronic pain. Patient-, provider-, and systems-level interventions may be needed to reduce these disparities.


MALDI-TOF mass spectrometry analysis of proteins and lipids in Escherichia coli exposed to copper ions and nanoparticles.
Escherichia coli (E. coli) is one of the most important foodborne pathogens to the food industry responsible for diseases as bloody diarrhea, hemorrhagic colitis and life-threatening hemolytic-uremic syndrome. For controlling and eliminating E. coli, metal nano-antimicrobials (NAMs) are frequently used as bioactive systems for applications in food treatments. Most NAMs provide controlled release of metal ions, eventually slowing down or completely inhibiting the growth of undesired microorganisms. Nonetheless, their antimicrobial action is not totally unraveled and is strongly dependent on metal properties and environmental conditions. In this work, we propose the use of matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry as a powerful tool for direct, time efficient, plausible identification of the cell membrane damage in bacterial strains exposed to copper-based antimicrobial agents, such as soluble salts (chosen as simplified AM material) and copper nanoparticles. E. coli ATCC 25922 strain was selected as 'training bacterium' to set up some critical experimental parameters (i.e. cell concentration, selection of the MALDI matrix, optimal solvent composition, sample preparation method) for the MS analyses. The resulting procedure was then used to attain both protein and lipid fingerprints from E. coli after exposure to different loadings of Cu salts and NPs. Interestingly, bacteria exposed to copper showed over-expression of copper binding proteins and degradation of lipids when treated with soluble salt. These findings were completed with other investigations, such as microbiological experiments. Copyright © 2016 John Wiley & Sons, Ltd.


Hydrogen scrambling in ethane induced by intense laser fields: statistical analysis of coincidence events.
Two-body Coulomb explosion processes of ethane (CH(3)CH(3)) and its isotopomers (CD(3)CD(3) and CH(3)CD(3)) induced by an intense laser field (800 nm, 1.0 × 10(14) W/cm(2)) with three different pulse durations (40 fs, 80 fs, and 120 fs) are investigated by a coincidence momentum imaging method. On the basis of statistical treatment of the coincidence data, the contributions from false coincidence events are estimated and the relative yields of the decomposition pathways are determined with sufficiently small uncertainties. The branching ratios of the two body decomposition pathways of CH(3)CD(3) from which triatomic hydrogen molecular ions (H(3)(+), H(2)D(+), HD(2)(+), D(3)(+)) are ejected show that protons and deuterons within CH(3)CD(3) are scrambled almost statistically prior to the ejection of a triatomic hydrogen molecular ion. The branching ratios were estimated by statistical Rice-Ramsperger-Kassel-Marcus calculations by assuming a transition state with a hindered-rotation of a diatomic hydrogen moiety. The hydrogen scrambling dynamics followed by the two body decomposition processes are discussed also by using the anisotropies in the ejection directions of the fragment ions and the kinetic energy distribution of the two body decomposition pathways.


Reciprocal connections between the claustrum and the gyrus sigmoideus posterior in the cat. An experimental study using the antegrade degeneration methods and the HRP retrograde axonal transport.
Following coagulation lesions affecting the gyrus sigmoideus posterior we investigated the termination field of the cortico-claustral projection using impregnation techniques (NAUTA, GYGAX 1954; FINK, HEIMER 1967). Preterminal and terminal fragments of the degenerating axons were identified ipsilaterally on the dorsal and lateral margins of the claustrum dorsale in the rostral half of the nucleus. Labeled neurons were found ipsilaterally within the claustrum dorsale, in the termination field of the cortico-claustral projection rising from the gyrus sigmoideus posterior following injections of HRP (Sigma VI) into the gyrus sigmoideus posterior. There are reciprocal connections between the dorsolateral region of the claustrum dorsale and the cortex of the gyrus sigmoideus posterior. A prevailing number of the HRP-positive neurons of the claustrum are located in the termination field of the particular cortico-claustral projection. The discussion deals with some problems concerning the cortico-claustral and claustro-cortical projections and presents opinions as to the functional significance of the claustrum.


Laparoscopic surgery for an esophageal duplication cyst using a near-infrared indocyanine green fluorescence system: A case report.
We herein describe a case of laparoscopic surgery for an esophageal duplication cyst using a near-infrared indocyanine green fluorescence system. A 64-year-old woman with a cystic tumor adjacent to the esophagogastric junction was referred to our hospital for treatment. Esophagogastroduodenoscopy and abdominal CT revealed a 70-mm submucosal tumor derived from the abdominal esophagus. We performed laparoscopic resection and then evaluated the tissue perfusion of the abdominal esophagus by using a near-infrared indocyanine green fluorescence system. A Dor fundoplication was performed to prevent postoperative gastroesophageal reflux disease and reinforce the mucosal layer defect. The postoperative course was uneventful, and pathological evaluation confirmed that the tumor was an esophageal duplication cyst. The patient did not develop recurrence in the 24 months after surgery. We have demonstrated that laparoscopic resection of an esophageal duplication cyst may be performed effectively with intraoperative assessment of tissue perfusion using a near-infrared indocyanine green fluorescence system.


Corticotrophin-releasing hormone in lipopolysaccharide-stimulated term fetal membranes and amniotic fluid from term and preterm birth in African Americans and Caucasians.
The objective of this study is to document differences in corticotrophin-releasing hormone (CRH), CRH receptor 1 (CRHR1), and CRH binding protein (CRHBP) gene expression in fetal membranes derived from African Americans and Caucasians in vitro in response to lipopolysaccharide (LPS) stimulation and to assess racial disparity in CRH concentrations in the amniotic fluid (AF) of women with spontaneous preterm birth (PTB). Fetal membranes (African American, n = 8; Caucasian, n = 8) at term, placed in an organ explant system, were stimulated with LPS. Microarray analysis documented differences in the mRNA expression pattern of CRH, CRHBP, and CRHR1 between races. CRH was measured in AF (a case [PTB]-control [term] study) and culture media. Between races, LPS significantly increased CRH and CRHR1 expression in African Americans and CRHBP in Caucasians, with no differences in controls. CRH was detectable only in LPS-stimulated African American membranes. AF CRH concentrations were higher in PTB compared with controls (P < .001), and no difference was noticed between races (P = .1). AF analysis did not document racial disparity in CRH concentrations in PTB. In fetal membranes, African Americans showed a higher expression and production of CRH in response to an in vitro stimulus.


[Contribution of Western blotting to the diagnosis of hydatidosis].
The aim of this study is to evaluate the contribution of the immunoWesternblot for the diagnosis and the post surgical follow-up of the hydatidosis. 71 sera from patients with hydatidosis confirmed by surgery were studied. All had a negative hydatic serology by screening tests (enzyme-linked immunosorbent assay, hemagglutination, electrosyneresis). 12 patients with sera in pre and post operative were monitored for 2 years. The Echinococcus Western blot IgG permitted to rectify the diagnosis of hydatidosis in 67.6 %. The rate of positivity was 100 % for the multivesicular liver cysts, 60 % for the young cysts and 50 % for the calcified cysts. Western blot permitted to rectify the diagnosis of lung cysts in 62.5 % of cases and in 50 % of cranial-spinal localizations. Analysis of Western Blot evolution in the 12 patients followed in pre and post-surgical revealed the disappearance of the bands 16, 18 and 26-28kDa in 8 month in the 8 patients with complete exeresis. This study proved the value added of Western blot compared to the other traditional techniques for the immunodiagnostic and the post-surgical monitoring of hydatidosis.


Quantitative Study of Using Piloti for Passive Climate Adaptability in a Hot-Summer and Cold-Winter City in China.
There has been an insufficient study of passive climate adaptability that considers both the summer and winter season for the outdoor thermal environment of hot-summer and cold-winter cities. In this study, we performed a quantitative simulation to research the passive climate adaptability of a residential area, considering piloti as the main method for climate adaptation in a hot-summer and cold-winter city in China. Numerical simulations were performed with a coupled simulation method of convection, radiation, and conduction. A cubic non-linear k⁻ε model proposed by Craft et al. was selected as the turbulence model and three-dimensional multi-reflections of shortwave and longwave radiations were considered in the radiation simulation. Through the simulation, we found that setting the piloti at the two ends of the building was the optimal piloti arrangement for climate adaptation. Then the relationship between the piloti ratio (0%, 20%, 40%, 60%, and 80%) and the outdoor thermal environment was studied. It could be concluded that with the increasing piloti ratio, the wind velocity increased, the mean radiant temperature (MRT) decreased slightly, and the average standard effective temperature (SET*) decreased to 3.6 °C in summer, while in winter, with the increasing piloti ratio, the wind velocity, MRT, and SET* changed slightly. The wind environment significantly affected the SET* value, and the piloti ratio should be between 12% and 38% to avoid wind-induced discomfort.


Second-generation antipsychotics and risk of cerebrovascular accidents in the elderly.
Concern has been recently raised for risperidone and olanzapine, possibly associated with cerebrovascular events in placebo-controlled trials conducted in elderly subjects with dementia. We investigated the relationship between exposure to second-generation antipsychotics (SGAs) and occurrence of cerebrovascular accidents in the elderly. From the regional database of hospital admissions of Lombardy, Italy, we extracted all patients aged 65 or older with cerebrovascular-related outcomes for the year 2002. From the regional database of prescriptions reimbursed by the National Health Service, we extracted all patients aged 65 or older who received antipsychotic prescriptions during 2001. The 2 databases were linked anonymously using the individual patient code. The proportions of cerebrovascular accidents were 3.31% (95% confidence interval, 2.95-3.69) in elderly subjects exclusively exposed to SGAs and 2.37% (95% confidence interval, 2.19-2.57) in elderly subjects exclusively exposed to first-generation antipsychotics. After background group differences were controlled for, exposure to SGAs significantly increased the risk of accidents. The analysis of cerebrovascular events in elderly subjects exposed to each individual SGA, in comparison with exposure to haloperidol, showed a significantly increased risk for risperidone only (adjusted odds ratio, 1.43; 95% confidence interval, 1.12-1.93). These data provide preliminary epidemiological evidence that exposure to SGAs, in comparison with exposure to first-generation antipsychotics, significantly increased the risk of cerebrovascular accidents in the elderly.


New pharmacologic options in the treatment of acute coronary syndromes and myocardial ischemia-reperfusion injury: potential role of levosimendan.
Modern and effective therapeutic possibilities have improved the management and outcomes in acute coronary syndromes and acute myocardial infarction. However, substantial morbidity and mortality still remain. Myocardial ischemia-reperfusion injury may contribute to additional damage to myocardial necrosis and apoptosis. Therefore, it has been focused on attention and field of therapeutic actions in the last years. The main mechanisms involved in the pathogenesis of ischemia-reperfusion injury are depressed energy metabolism, elevated oxidative damage, and altered calcium homeostasis. In experimental trials, a variety of drugs have proved effectiveness for the prevention and treatment of the ischemia-reperfusion injury. However, its efficacy, not always confirmed, has not yet been established in clinical practice. On the basis of the strong evidence linking potassium ATP dependent channels opening in the myocardium and its proved cardioprotective role during ischemia, these channels have been pointed out as possible and promising pharmacological targets in this setting. Some evidences suggest that the calcium sensitizing agent levosimendan may have of beneficial and exerts cardioprotective effects on myocardial ischemia and ischemia-reperfusion injury. Further investigation is warranted on this novel application of levosimendan.


Evaluation of impression smear in the diagnosis of fungal keratitis.
To evaluate and compare impression smear with the conventional mechanical corneal scraping by potassium hydroxide (KOH) method in the diagnosis of fungal keratitis. Fifty consecutive cases of clinically suspected fungal keratitis were included. Under topical anesthesia, impression smear was obtained from the lesion, followed by the conventional mechanical scraping and both were transferred to sterile glass slides. The slides were examined under direct microscopy with KOH 10% wet mount. Impression smear KOH was compared with the scraping KOH and culture. The KOH smear was positive in 36 (72%) patients with impression smear technique and positive in 35 (70%) patients from corneal scrapings. Sensitivity of impression smear technique as compared to scraping was 97.14%, specificity 92.86%, positive predictive value 94.4%, negative predictive value 92.86%, false positive rate 5.56%, false negative rate 7.14% and accuracy was 94%. As compared to culture, the sensitivity was 100%, specificity 46.67%, positive predictive value 55.56%, negative predictive value 100%, false positive rate 53.33%, false negative rate 0% and accuracy was 68%. Impression smear KOH examination is comparable to conventional mechanical corneal scraping KOH examination in making a tentative diagnosis of fungal keratitis and can be accurately relied upon for initiating anti-fungal therapy.


Persistent draining crossover in DNA and other semi-flexible polymers: Evidence from hydrodynamic models and extensive measurements on DNA solutions.
Although the scaling theory of polymer solutions has had many successes, this type of argument is deficient when applied to hydrodynamic solution properties. Since the foundation of polymer science, it has been appreciated that measurements of polymer size from diffusivity, sedimentation, and solution viscosity reflect a convolution of effects relating to polymer geometry and the strength of the hydrodynamic interactions within the polymer coil, i.e., "draining." Specifically, when polymers are expanded either by self-excluded volume interactions or inherent chain stiffness, the hydrodynamic interactions within the coil become weaker. This means there is no general relationship between static and hydrodynamic size measurements, e.g., the radius of gyration and the hydrodynamic radius. We study this problem by examining the hydrodynamic properties of duplex DNA in solution over a wide range of molecular masses both by hydrodynamic modeling using a numerical path-integration method and by comparing with extensive experimental observations. We also considered how excluded volume interactions influence the solution properties of DNA and confirm that excluded volume interactions are rather weak in duplex DNA in solution so that the simple worm-like chain model without excluded volume gives a good leading-order description of DNA for molar masses up to 10(7) or 10(8) g/mol or contour lengths between 5 μm and 50 μm. Since draining must also depend on the detailed chain monomer structure, future work aiming to characterize polymers in solution through hydrodynamic measurements will have to more carefully consider the relation between chain molecular structure and hydrodynamic solution properties. In particular, scaling theory is inadequate for quantitative polymer characterization.


Order and chaos in the rotation and revolution of a line segment and a point mass.
We study the classical dynamics of two bodies, a massive line segment or slash (/) and a massive point or dot (.), interacting gravitationally. For this slash-dot (/.) body problem, we derive algebraic expressions for the force and torque on the slash, which greatly facilitate analysis. The diverse dynamics include a stable synchronous orbit, generic chaotic orbits, sequences of unstable periodic orbits, spin-stabilized orbits, and spin-orbit coupling that can unbind the slash and dot. The extension of the slash provides an extra degree of freedom that enables the interplay between rotation and revolution. In this way, the slash-dot body problem exhibits some of the richness of the three body problem with only two bodies and serves as a valuable prototype for more realistic systems. Applications include the dynamics of asteroid-moonlet pairs and asteroid rotation and escape rates.


Heparin mapping using heparin lyases and the generation of a novel low molecular weight heparin.
Seven pharmaceutical heparins were investigated by oligosaccharide mapping by digestion with heparin lyase 1, 2, or 3, followed by high performance liquid chromatography analysis. The structure of one of the prepared mapping standards, ΔUA-Gal-Gal-Xyl-O-CH(2)CONHCH(2)COOH (where ΔUA is 4-deoxy-α-l-threo-hex-4-eno-pyranosyluronic acid, Gal is β-d-galactpyranose, and Xyl is β-d-xylopyranose) released from the linkage region using either heparin lyase 2 or heparin lyase 3 digestion, is reported for the first time. A size-dependent susceptibility of site cleaved by heparin lyase 3 was also observed. Heparin lyase 3 acts on the undersulfated domains of the heparin chain and does not cleave the linkages within heparin's antithrombin III binding site. Thus, a novel low molecular weight heparin (LMWH) is afforded on heparin lyase 3 digestion of heparin due to this unique substrate specificity, which has anticoagulant activity comparable to that of currently available LMWH.


Induction of virus-specific immunity by iscoms.
Immune stimulating complexes (iscoms) are 30-40 nm cage-like structures, which consist of glycoside molecules of the adjuvant Quil A, cholesterol and phospholipids in which antigen can be integrated. Even in the presence of pre-existing antibodies they may function as a potent adjuvant system by inducing high systemic and local long-lasting antibody and T cell responses. An additional advantage is their capacity to induce MHC class I restricted CD8+ CTL responses. This combination will in most cases, when the relevant antigens are incorporated, lead to adequate protection against virus infection or disease. An overview is given of the present data available on the use of iscoms in viral systems, and some relevant examples are discussed to highlight general principles in the use of iscoms as an adjuvant system.


[The influence of subclinical level of graft versus host reaction on reconstitution of host lymphoid tissues].
In this study reconstitution of lymphoid tissues under the influence of subclinical graft versus host reaction (GVHR) was investigated. Lethally irradiated recipient mice (AKR: H-2k, Mls-1a) were reconstituted with donor (B10: H-2b, Mls-1b) bone marrow (BM) cells treated with anti-Thy 1 antibody alone without complement (GVHR chimeras). When their immunological reconstitution was analysed and compared with that of AKR recipients which had been reconstituted with B10 BM treated with anti-Thy 1 and complement (Control chimera), both of these chimeras all survived more than 100 days without showing clinical signs of GVHD. Full donor chimerism was accomplished at an early stage after BM transplantation (BMT) in GVHR chimeras, whereas substantial number of recipient T cells persisted in Control chimeras during the entire observation period. No significant difference was observed in the reconstitution of the thymus and spleen between the Control and GVHR chimeras. By contrast, the reconstitution of lymph nodes (LN) in GVHR chimeras was inhibited. The apparent failure of LN reconstitution was attributed to damage of LN structure involved in homing of lymphocytes, but not attributable to damage of lymphocytes themselves. Since donor CD8+ T cells caused the damage, target antigens of this damage appeared to be H-2 class I molecules in the present system. Furthermore, clonal deletion of V beta 6+T cells that are reactive to recipient Mls-1a antigen was induced in Control chimeras but abrogated in the GVHR chimeras. The latter abrogation of clonal deletion of V beta 6+T cells appeared to result from the early disappearance of recipient T cells in these GVHR chimeras.


A condition metric for Eucalyptus woodland derived from expert evaluations.
The evaluation of ecosystem quality is important for land-management and land-use planning. Evaluation is unavoidably subjective, and robust metrics must be based on consensus and the structured use of observations. We devised a transparent and repeatable process for building and testing ecosystem metrics based on expert data. We gathered quantitative evaluation data on the quality of hypothetical grassy woodland sites from experts. We used these data to train a model (an ensemble of 30 bagged regression trees) capable of predicting the perceived quality of similar hypothetical woodlands based on a set of 13 site variables as inputs (e.g., cover of shrubs, richness of native forbs). These variables can be measured at any site and the model implemented in a spreadsheet as a metric of woodland quality. We also investigated the number of experts required to produce an opinion data set sufficient for the construction of a metric. The model produced evaluations similar to those provided by experts, as shown by assessing the model's quality scores of expert-evaluated test sites not used to train the model. We applied the metric to 13 woodland conservation reserves and asked managers of these sites to independently evaluate their quality. To assess metric performance, we compared the model's evaluation of site quality with the managers' evaluations through multidimensional scaling. The metric performed relatively well, plotting close to the center of the space defined by the evaluators. Given the method provides data-driven consensus and repeatability, which no single human evaluator can provide, we suggest it is a valuable tool for evaluating ecosystem quality in real-world contexts. We believe our approach is applicable to any ecosystem.


High speed optical holography of retinal blood flow.
We performed noninvasive video imaging of retinal blood flow in a pigmented rat by holographic interferometry of near-infrared laser light backscattered by retinal tissue, beating against an off-axis reference beam sampled at a frame rate of 39 kHz with a high throughput camera. Local Doppler contrasts emerged from the envelopes of short-time Fourier transforms and the phase of autocorrelation functions of holograms rendered by Fresnel transformation. This approach permitted imaging of blood flow in large retinal vessels (∼30 microns diameter) over 400×400 pixels with a spatial resolution of ∼8 microns and a temporal resolution of ∼6.5 ms.


Pipeline to Military Orthopaedic Leadership: 20 Years of Race and Gender Diversity Trends Within Military Orthopaedic Surgery Fellowship Training.
The lack of trainees from underrepresented race and gender backgrounds in orthopaedic surgery fellowship training has been well reported in the literature. The purpose of this study was to investigate the demographic trends of federally sponsored military orthopaedic surgery fellows in the Army, Navy, and Air Force. We hypothesize that there has been an increase in women selected for fellowship but that there has been no change in the race demographics of military fellows over the past 2 decades. A retrospective review of all available demographic data collected by the Army, Air Force, and Navy since the beginning of tracking federally funded fellowship training in orthopaedic surgery was completed (1998-2021). Data were grouped into 4-year periods for analysis to closely mirror the military assignment cycle. Three hundred sixty-two military orthopaedic surgery fellowship board selectees were included in our analysis. The proportion of women fellows increased from 3% (n = 2/69) over 2001 to 2004 to 21% (n = 17/82) during 2017 to 2020 (p < 0.05). Fellows who identified as White comprised 82% (n = 297) of the cohort during the study period. Individuals who identified as Asian were the next highest proportion of fellows at 4% (n = 16), followed by Black (n = 14, 4%) and Hispanic (n = 13, 3%). Individuals who identified as Native Hawaiian/Pacific Islander represented 1% (n = 3), and an additional 6% (n = 20) fellows identified as "other" or "undeclared." Over the 20-year study period, representation of Asian, Black, Native Hawaiian, and Hispanic fellows did not increase (p = 0.79, 0.81, 0.45, 0.34, respectively). Within military orthopaedics, there has been increased representation of women in fellowship training over the past 20 years. However, the proportion of fellows from underrepresented racial and ethnic groups has remained stagnant. One barrier to improving gender and race representation is the currently imprecise and inconsistent collection of demographic information. Importantly, fellowship training has a direct effect on future leadership opportunities within the military orthopaedic surgery community. A more diverse leadership may help to inspire future generations of military orthopaedic surgeons. IV.


Contribution of Ultrasonography in the Diagnosis of Periportal Fibrosis Caused by Schistosomiasis.
Periportal fibrosis is one of the major complications of schistosomiasis infection. Specific images of this infection revealed by ultrasonography allow the assessment of the different stages. Our purpose is to describe the ultrasonographic appearances of periportal fibrosis due to schistosomiasis infection. The study was retrospective descriptive in the Medical Imagery Centre of CHUJRA. A total of 29 patients showed upper and/or lower digestive hemorrhage and a positive result in schistosomiasis serology. The median age was 41.2 years. Male gender dominated with 54.7%. In 78.3% of the cases, the patients were farmers in schistosomiasis-endemic areas. On ultrasonography, 81.3% were found to present Stage III periportal fibrosis, 11.5% presented Stage II, and 7.2% presented Stage I. There was no case of portal thrombosis. Splenomegaly was found in 83.2% of the cases and hepatomegaly in 48.9% of the cases. Porto-systemic shunt was detected in 80.7% of the cases and ascites in 72.3%. Ultrasonography examination represents an important role in the diagnosis of periportal fibrosis, in its staging process, and allows the assessment of porto-systemic, hepatic, and splenic alterations.


Investigations on factors affecting chitosan for dissolution enhancement of oxcarbazepine by spray dried microcrystal formulation with an experimental design approach.
In the present work effect of chitosan on microcrystal formulation for dissolution enhancement of oxcarbazepine using controlled crystallization technique coupled with spray drying was explored. The work was extended for exploration of simplified approach for stable particle size reduction. The study was performed with an experimental design approach i. e. a fractional factorial design of resolution 5 (with all 2 factor interaction) for the screening of predefined independent variables drug concentration, chitosan concentration, feed rate, inlet temperature and percent aspiration for spray drying. Whereas percent drug dissolved, wettability time, flowability in terms of angle of repose and particle size were designated as response variables. Resultant models were analyzed using multiple linear regression analysis, which generated equation to plot response surface curves along with desirability function. Results showed that chitosan concentration had significant effect on dissolution enhancement of oxcarbazepine at a level of 2% w/v. Increase in drug concentration showed decreased dissolution rate however on particle size it did not show statistically significant effect. Topographical characterization was carried out by SEM which showed that feed rate, percent aspiration and inlet temperature had significant effect on particle morphology. For deriving optimized formulation results were analyzed using desirability function for the maximum percent drug dissolved and least drug polymer matrix particle size. DSC studies showed that drug was molecularly associated with chitosan matrix or particles.


A comparison of perceptions of quality of life among adults with spinal cord injury in the United States versus the United Kingdom.
To identify which aspects of life are most important to adults with spinal cord injury (SCI) and compare perspectives in the United States and the United Kingdom. We conducted 20 in-depth interviews with adults with SCI (ten in the US and ten in the UK). Verbatim transcriptions were independently analyzed line-by-line by two coders using an inductive approach. Codes were grouped into themes about factors that constitute and affect quality of life (QOL). Five overarching themes emerged: describing QOL in the context of SCI; functional adjustment; medical care; financial resources; and socio-political issues. Twenty subthemes emerged on factors that affect QOL. Participants in both samples identified medical care as a key influence on QOL. The US group talked about a predominantly negative influence (e.g., fragmented primary and specialist care, insurance constraints, bureaucracy), whereas UK interviewees mentioned a predominantly positive influence (e.g., universal provision, including free and continuous care, free wheelchairs and home care, and length of rehabilitation commensurate with level of injury). Functional adjustment, such as physical and mental adjustment post-discharge and aging with SCI, was another important contributor to QOL, and varied by country. Most US interviewees reported poor knowledge about self-care post-discharge and poor quality of home adaptations compared to the UK group. For adults living with SCI, good QOL is essential for successful rehabilitation. Differences between interviewees from the two countries in perceived medical care and functional adjustment suggest that factors affecting QOL may relate to broader health system characteristics.


Effects of proton beam irradiation on mitochondrial biogenesis in a human colorectal adenocarcinoma cell line.
Proton beam therapy has recently been used to improve local control of tumor growth and reduce side-effects by decreasing the global dose to normal tissue. However, the regulatory mechanisms underlying the physiological role of proton beam radiation are not well understood, and many studies are still being conducted regarding these mechanisms. To determine the effects of proton beams on mitochondrial biogenesis, we investigated: mitochondrial DNA (mtDNA) mass; the gene expression of mitochondrial transcription factors, functional regulators, and dynamic-related regulators; and the phosphorylation of the signaling molecules that participate in mitochondrial biogenesis. Both the mtDNA/nuclear DNA (nDNA) ratio and the mitochondria staining assays showed that proton beam irradiation increases mitochondrial biogenesis in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced aggressive HT-29 cells. Simultaneously, proton beam irradiation increases the gene expression of the mitochondrial transcription factors PGC-1α, NRF1, ERRα, and mtTFA, the dynamic regulators DRP1, OPA1, TIMM44, and TOM40, and the functional regulators CytC, ATP5B and CPT1-α. Furthermore, proton beam irradiation increases the phosphorylation of AMPK, an important molecule involved in mitochondrial biogenesis that is an energy sensor and is regulated by the AMP/ATP ratio. Based on these findings, we suggest that proton beam irradiation inhibits metastatic potential by increasing mitochondrial biogenesis and function in TPA-induced aggressive HT-29 cells.


Effect of ciprofibrate on C-reactive protein and fibrinogen levels.
Statins can lower the circulating levels of C reactive protein (CRP). This effect may be relevant because CRP is a predictor of vascular risk. In contrast, the evidence that fibrates lower CRP levels is very limited. The effect of treatment with ciprofibrate (100 mg once daily) was investigated for 8 weeks in 30 patients with primary dyslipidemia. There was a significant (p < 0.01) decrease in median (range) CRP levels by 36.8% from 1.9 mg/L (1.0-6.0 mg/L) to 1.2 mg/L (1.0-5.5 mg/L). Plasma fibrinogen levels were also significantly (p = 0.05) reduced. There was no correlation between the fall in CRP levels and the changes in lipid or fibrinogen levels. These findings support the concept that fibrates, like the statins, lower serum CRP levels. However, fibrates have a different mode of action. Fibrates (with the exception of gemfibrozil) also consistently lower plasma fibrinogen levels. In contrast, the effect of statins on the circulating levels of this coagulation factor remains to be defined. These differences may help in defining the mechanisms responsible for drug-induced changes in the circulating levels of CRP and fibrinogen. A favorable effect on CRP and fibrinogen levels may increase the clinical efficacy of statins and fibrates.


Effects of heat shock on the production of human erythropoietin from recombinant CHO cells.
The production of recombinant proteins by mammalian cells demands a highly controlled environment for cell cultivation. Temperature stress represents a commonly encountered disturbance in both research and process environments. In this study, we examined the effects of heat shock on the expression of recombinant human erythropoietin (EPO) in a Chinese hamster ovary (CHO) cell line. Biosynthetic radiolabeling experiments indicated that the cells exposed to a 42 degrees C/1-hour heat shock exhibit a transient reprogramming of transcription and translation characterized by the inhibition of protein synthesis and induction of heat shock proteins. The rate of protein synthesis decreased by 50% after the heat shock, while the rate of RNA synthesis increased by 50% initially and then quickly reduced to 80% of the control level. The protein and RNA synthesis rates were fully recovered in approximately 48 hours after the heat shock. However, we found that the expression of EPO was not arrested by the heat shock. The glycosylation patterns, as examined by isoelectric focusing, of either the culture supernatant or the purified EPO were not affected by the heat shock. In contrast, a 45 degrees C/1-hour heat shock terminated RNA and protein synthesis immediately and caused culture death in 12 hours. Cellular responses to temperature stress were affected by the metabolic state of the cells; cells maintained in serum-free medium were more sensitive than cells growing exponentially in the presence of serum. We have also examined the kinetics of metabolic responses of the cells to heat shock with respect to nutrient uptake and metabolite accumulation.


The endosperm-specific expression of a rice prolamin chimaeric gene in transgenic tobacco plants.
The 5' upstream region (-680 to +40), containing the potential promoter and complete signal peptide coding sequence of the rice seed storage prolamin gene was amplified in vitro using the polymerase chain reaction from the genome of Chinese rice cultivar Zhonghua 8. The physical map and DNA sequence analysis show strong homology with the 5' flanking region of the rice prolamin gene published by Kim and Okita (1988a). No change in the signal peptide coding sequence and a long leader sequence with several small open reading frames were found. The chimaeric gene containing the 5' flanking region of the prolamin gene (-680 to -18) was transcriptionally fused with the beta-glucuronidase (GUS) reporter gene and the fusion junction was confirmed by both physical mapping and DNA sequence analysis. The resultant chimaeric gene was used to transform tobacco explants, using the Ti binary system of Agrobacterium tumefaciens LBA4404. Three transgenic tobacco plants with as many as 20 copies of the chimaeric GUS gene (confirmed by dot and Southern hybridization) were analysed further. Histochemical analysis revealed GUS activity in the endosperm tissue of tobacco seed at the developmental stage about 20 days after flowering (DAF). No GUS activity was found in leaves, stems, roots and flowers of the transgenic tobacco plants. Therefore, we conclude that the 5' upstream region from -680 to -18 was sufficient to confer the endosperm-specific expression of the rice prolamin gene.


Distal tibial physeal fractures in children that may require open reduction.
Fractures of the distal end of the tibia in children often involve the physis. They are of particular importance because partial growth arrest can occur and result in angular deformity, limb-length discrepancy, or incongruity of the joint surface (or a combination of these). We evaluated the cases of thirty-two children who had a fracture leading to established partial growth arrest of the distal end of the tibia. Most of this group had had a Salter-Harris Type-III or Type-IV fracture. Twenty-eight of the fractures had been treated by gentle closed reduction and immobilization in a plaster cast. We also evaluated the cases of thirty-three children who were seen by us for treatment of an acute fracture; most of these were Salter-Harris Type-III or Type-IV fractures of the distal end of the tibia. Nineteen of the twenty acute Type-III or Type-IV fractures that were treated with accurate open reduction of the physis and internal fixation healed without growth disturbance, while five of the nine fractures that were treated by closed means formed a bone bridge, presaging a disturbance in growth. This study suggests that Salter-Harris Type-III and Type-IV, and perhaps Type-II, fractures of the distal end of the tibia commonly cause disturbance of growth in the tibia, and that anatomical reduction of the physis by closed or open means may decrease the incidence of these disturbances of growth, including shortening and varus angulation of the ankle.


Dermatology services: The new normal post COVID-19.
The article discusses the impact of COVID-19 on the future of dermatology services. It will explore the changes dermatology services may need to follow to minimise disease transmission. This will include an integration of teledermatology into everyday practice, a shift in dermatology training, and change in dermoscopy technique.