Extracranial complications of severe head injury.
In order to define the role of intracranial and extracranial complications in determining outcome from severe head injury, 734 patients from the Traumatic Coma Data Bank were analyzed. Nine classes of intracranial and 13 classes of extracranial complications occurring within the first 14 days after admission were analyzed, while controlling for age, admission Glasgow Coma Scale motor score, early hypoxia or hypotension, and severe extracranial trauma. Outcome for survivors was based on the last recorded Glasgow Outcome Scale score, obtained a median of 521 days after injury. Intracranial complications did not significantly alter outcome for the study group. Of the extracranial complications, pulmonary, cardiovascular, coagulation, and electrolyte disorders occurred most frequently at 2 to 4 days. Infections developed later, peaking at 5 to 11 days. Gastrointestinal, renal, and hepatic complications followed no specific time course. Electrolyte abnormalities were the most frequent occurrence (59% of patients) but did not alter outcome. Pulmonary infections (41%), shock (29%, systemic blood pressure < or = 90 mm Hg for 30 minutes or more), coagulopathy (19%), and septicemia (10%) were significant independent predictors of an unfavorable outcome. Backward-elimination, stepwise logistic regression modeling indicated that the estimated reduction of unfavorable outcome was 2.9% for the elimination of pneumonia, 3.1% for coagulation disturbances, 1.5% for septicemia, and 9.3% for shock. These data suggest that extracranial complications are highly influential in determining the outcome from severe head injury and that significant improvements in outcome in a sizeable proportion of patients could be accomplished by improving the ability to prevent or reverse pneumonia, hypotension, coagulopathy, and sepsis.


Clinical Performance of Direct Posterior Composite Restorations in Patients with Amelogenesis Imperfecta.
The objective of this study was to analyze and assess the clinical performance of direct composite restorations using a nanohybrid and a nanofill composite material for posterior teeth in patients with amelogenesis imperfecta (AI). This study involved 15 patients between the ages of 14 and 30 years suffering from amelogenesis imperfecta (AI). During the study, the patients received direct composite restorations using either the Clearfil Majesty ES-2 (Kuraray Medical Inc, Tokyo, Japan) and Clearfil Universal Bond (Kuraray) or Filtek Ultimate Universal Restorative (3M ESPE, St Paul, MN, USA) and Single Bond Universal Adhesive (3M ESPE). The evaluations of the restorations were conducted per the modified USPHS criteria at the time of baseline as well as during the first-, second-, third-, and fourth-year follow-up sessions. After four years, it was observed that the cumulative success rate of direct posterior restorations was 98.1% for Clearfil Majesty ES-2 and 92.2% for Filtek Ultimate. During the study one Clearfil Majesty ES-2 restoration and four Filtek Ultimate restorations failed. There was a significant difference between Clearfil Majesty ES-2 and Filtek Ultimate in the color match in posterior restorations after three and four years. The causes of failure included marginal discoloration and caries, as well as fracture of the restoration. Hence, it can be stated that the use of nanohybrid or nanofill composites in posterior direct restorations in patients with AI looks promising. The failure rate of Clearfil Majesty ES-2 was found to be lower than that of Filtek Ultimate restorations. Clinically, the rate of optimum restorations conducted for partial discoloration, marginal adaptation, color match, and surface texture were observed to be higher when Clearfil Majesty ES-2 was used. However, additional studies are needed to assess the clinical performance of direct posterior composite materials in patients with AI.


Theophylline blocks ethanol withdrawal-induced hyperalgesia.
This study examined the effects of theophylline on the hyperalgesia produced by ethanol withdrawal using a radiant heat tail-flick assay. Chronic effects of ethanol were tested in four groups of rats which received 10 days exposure to a liquid diet [ethanol alone or with theophylline [0.5 and 1.0 mg/kg, twice daily, intraperitoneally (i.p.)], and dextrin control diet]. Ethanol withdrawal was tested 12 h after removal of the liquid diet. Effects of cumulative doses of the non-selective adenosine agonist 2-chloroadenosine (2-CADO; 0.6-10 mg/kg, i.p.) were tested during withdrawal in the ethanol-treated groups. Chronic exposure to ethanol produced antinociception, and hyperalgesia was seen during withdrawal. Subchronic administration of theophylline (0.5-1.0 mg/kg, twice daily, i.p.) dose-dependently prevented the ethanol-withdrawal-induced hyperalgesia. During ethanol withdrawal, 2-CADO was less potent than when given to non-dependent rats and this effect was prevented by subchronic administration of theophylline (1.0 mg/kg). These findings provide behavioural evidence in agreement with earlier work on the role of adenosine in the development of ethanol tolerance and withdrawal, and suggest that adenosine receptors play an important role in the development of hyperalgesia during ethanol withdrawal.


[Association between preventive care provided in public dental services and caries prevalence].
To assess the association between preventive care provided in public dental services and young people's oral health. Oral health data on 4,033 young people aged 15 to 19 years living in 85 municipalities of the state of Rio Grande do Sul, Southern Brazil, were obtained from the national oral health survey "Saúde Bucal Brasil 2003" for the period 2002-2003. The following variables were studied: age, gender, income, education, time elapsed since last dental visit, reason for dental visit, and water fluoridation. Data on dental care services were obtained from the national database of public health services. Statistical analysis was performed using multilevel logistic regression. Youngsters from the 21 municipalities with the lowest preventive care (scaling + fluoride + sealants) rates per 100 inhabitants were 2.27 (95% CI: 1.45;3.56) more likely to have non-filled dental cavities than those from the 21 municipalities with the highest care rates. After adjustment for a number of individual and contextual factors this likelihood decreased to 1.76 (95% CI: 1.13;2.72). The variance attributable to variables at municipal level was 14.1% for the empty model and decreased to 10.5% for the fully adjusted model. Rio Grande do Sul public dental services may have contributed for the reduction in the number of non-filled cavities in young people. However, it was not possible to detect the impact of this service on total dental caries experience.


Reconstructing exposures from the UK chemical warfare agent human research programme.
The UK government has carried out a research programme studying military capability under conditions of chemical warfare at a facility at Porton Down, Wiltshire, since World War I. In 2001 the Ministry of Defence commissioned a cohort study to investigate the long-term health effects on military veterans of their participation in this programme. We assessed the availability and quality of exposure assessment data held in the archive at Porton Down for the purpose of this study. This involved looking in detail at exposure data in a sample of 150 veterans and undertaking a general review of all available records held in the archive. These sources suggested that the Porton Down records were largely complete and included sufficient identifying information for linkage with service personnel data and with national mortality and cancer registration records. Servicemen usually had multiple tests so data were most readily available in a test-wise format, allowing subsequent aggregation of tests by individual. The name of the chemical used in each test could be determined for most tests and most of the named chemicals could be categorized into major groups for epidemiological analyses. For the major groups (vesicants and nerve agents), quantitative data were available on exposure and on acute toxicity. Standardization will be required of the several different units which were used. Based on this study, exposure assessment for the cohort study of Porton Down veterans will involve abstraction of the name of the chemical used in each test, with quantitative data on exposure and acute toxicity for vesicants and nerve agents. Our results here show that experimental records at Porton Down offer a unique and valuable resource for reconstructing the chemical exposures used in this research programme. The resulting cohort study has the potential to provide information which will assist in understanding the long-term health impact of chemical warfare agent exposure on these veterans.


Emergency thoracotomy: appropriate use in the resuscitation of trauma patients.
The objective of this study was to evaluate the use of emergency thoracotomy in our institution in an effort to determine whether this procedure is both beneficial and cost effective in blunt and/or penetrating trauma. We conducted a retrospective review of charts and coroner's reports. Our setting was a Level I trauma center in a tertiary-care facility. We examined the cases of trauma patients presenting to the trauma center over a 2-year period. Of 2490 patients who presented to the emergency department over the study period 41 underwent early thoracotomy. Twelve of these were excluded from the study because their cases were not truly emergent. Of the remaining 29 ten were admitted for penetrating injuries and 19 for blunt injuries. The average Injury Severity Scores for penetrating and blunt injuries were 30 and 40 respectively. There were four blunt trauma patients who died in the emergency department, 15 went to the operating room, and five who survived to go to the intensive care unit. All blunt trauma patients requiring emergency thoracotomy died within 9 days of presentation. Of the ten penetrating wound patients two died in the emergency department, four died in the operating room, and four went to the intensive care unit after surgery. One of the four patients who went to the intensive care unit died approximately 6 days after injury. The other three patients survived and are now living normal productive lives. All survivors of penetrating trauma who required emergency thoracotomy had their procedure performed in the operating room. Overall survival rates for penetrating and blunt trauma were 30 and 0 per cent respectively. Pericardial tamponade was found in 50 per cent of the penetrating trauma patients (two of the three survivors) and four of 19 of the blunt trauma patients. This reinforces the importance of a prompt pericardiotomy upon opening the chest. At our institution the algorithm for emergency thoracotomy is liberal and is not cost effective for blunt trauma. We need to re-evaluate our decision-making process concerning the use of emergency thoracotomy especially in the blunt trauma patient. The review also shows the importance of pericardiotomy when performing an emergency thoracotomy.


Cell polarity protein Lgl2 is lost or aberrantly localized in gastric dysplasia and adenocarcinoma: an immunohistochemical study.
The diagnosis of gastric epithelial dysplasia, a precursor lesion of gastric adenocarcinoma, is hindered by interobserver variability and by its resemblance to regenerative changes. Loss of cell polarity, a histological feature of gastric epithelial dysplasia, may be difficult to ascertain, especially in the setting of inflammation or injury. A biomarker of cell polarity could be useful in diagnosis of dysplasia, but has not been reported. The Lethal giant larvae (lgl) gene controls apical-basal polarity of epithelial cells in Drosophila, and has properties of a tumor-suppressor gene. Two homologs, lgl1 and lgl2, are present in mammals and lgl2 mRNA is highly expressed in the stomach. The goal of our study was to test the hypothesis that Lgl2 protein expression and/or localization are disrupted in gastric epithelial dysplasia and adenocarcinoma. Routinely processed pathology specimens including 94 benign mucosae of digestive organs, in addition to 36 reactive gastropathy, 57 gastric epithelial dysplasia, and 77 gastric adenocarcinomas, were immunostained for Lgl2 protein. All normal, reactive, and chronically inflamed gastric epithelia showed basolateral Lgl2 staining. Normal esophageal, duodenal, colonic, biliary, and pancreatic duct mucosae, as well as gastric intestinal metaplasia, did not express Lgl2. All but one case each of gastric epithelial dysplasia and adenocarcinoma showed either complete loss of anti-Lgl2 immunoreactivity or diffuse, mostly weak, cytoplasmic staining. Complete loss of immunoreactivity was significantly more often observed in diffuse-type than in intestinal-type adenocarcinomas (79 vs 48%, respectively). Our data suggest that Lgl2 expression is either aberrantly localized or lost in gastric epithelial dysplasia and adenocarcinoma, whereas it is maintained in reactive gastric mucosa. We propose that Lgl2 may be a potential marker to rule out gastric epithelial dysplasia and adenocarcinoma in diagnostic specimens. However, the consistently negative anti-Lgl2 immunoreactivity seen in intestinal metaplasia does not allow differentiation of dysplasia from intestinal metaplasia with reactive change.


Internal exposure to (210)Po and (40)K from ingestion of cooked daily foodstuffs for adults in Japanese cities.
The isotope (210)Po was suspected of being involved in the death of a former Russian intelligence agent in 2006 in the UK. Although human exposure to this natural radionuclide in foods is estimated to be high, few studies are available. UNSCEAR Report 2000 does not contain data on (210)Po concentrations of foodstuffs in Japan. We analyzed samples of the everyday Japanese diet cooked with foodstuffs purchased at supermarkets in 7 major domestic cities in 2007-2008. (210)Po was quantified by alpha spectrometry and natural radionuclides such as (40)K by gamma spectrometry. The daily intake and committed effective dose of (210)Po, (40)K, and other natural radionuclides for Japanese adults were calculated. Daily intake was 0.34-1.84 (mean +/- sigma : 0.66 +/- 0.53) and 68.5-94.2 (81.5 +/- 8.5) Bq/d and the committed effective dose was 0.15-0.81 (0.29 +/- 0.24) and 0.16-0.21 (0.18 +/- 0.02) mSv for (210)Po and (40)K, respectively, comprising a high percentage of the total exposure. The total of the mean committed effective dose for the two nuclides (0.47 mSv) was higher than the annual effective dose from ingestion of foods reported by UNSCEAR 2000 (0.29 mSv). The mean committed effective dose of (40)K in the 7 major Japanese cities was comparable to the global average (0.17 mSv). The dietary exposure of Japanese adults can be characterized by a higher (210)Po contribution than in other countries. Of the total daily dietary (210)Po exposure (13 food categories excluding water) for adults in Yokohama, about 70% was from fish/shellfish and 20% from vegetables/mushrooms/seaweeds, reflecting preferences of Japanese to eat a considerable amount of fish/shellfish containing high (210)Po concentrations.


Safety and persistence of non-animal stabilized hyaluronic Acid fillers for nasolabial folds correction in 30 Indian patients.
Correction of nasolabial creases through minimally invasive procedures is increasingly being sought by patients. Injecting non-animal stabilized hyaluronic acid filler is a highly effective method to achieve an optimal and persistent cosmetic result To evaluate the efficacy, persistence and safety of Restylane and Perlane (Q-Med, Sweden) for correction of nasolabial folds in Indian patients Thirty Indian patients with mild, moderate and severe nasolabial folds (based on Wrinkle Assessment Scale) were recruited in the study after informed consent for correction of their folds with Restylane or Perlane or both. Injections were administered in a single sitting after global assessment of the patient's face using Wrinkle assessment scale (WAS).Optimal filling was performed by using appropriate techniques and its safety and efficacy assessed independently by the investigator as well as by patients at immediately, 3, 6 and 9 months post-procedure. Any adverse reactions were noted. Twenty two females and 8 males (age range 45-55 years, mean age 52 years) were recruited in the study. An optimum cosmetic correction was obtained in all patients. The efficacy increased with time and was greatest at 3 months after the treatment. Grade 2 improvement was maintained at 9 months in mild and moderate folds, and grade 3 improvement for severe folds. Minor post injection side effects like erythema at puncture site, needle marks and bruising were seen. Restylane and Perlane are safe and effective dermal fillers for correction of nasolabial creases and offer immediate effect.


Cardiac magnetic resonance imaging for the assessment of the myocardium after doxorubicin-based chemotherapy.
Doxorubicin is associated with a cumulative dose-dependent nonischemic cardiomyopathy. Cardiac magnetic resonance imaging (cMRI) is able to examine both structural and functional components of the myocardium. Our aim was to assess the myocardial changes in non-Hodgkin lymphoma patients undergoing doxorubicin-based chemotherapy using cMRI. cMRI examination was performed before and 3 months after chemotherapy. Experienced investigators interpreted each cMRI, and were blinded to all data. Left ventricular ejection fractions (LVEF), cardiac deformation, and delayed gadolinium enhancement (GD-DE) were quantified for each cMRI. The change between LVEF, GD-GE, and cardiac deformation parameters were compared between the 2 cMRI studies. A Δ LVEF≥10% was considered clinically relevant. The findings of GD-GE or changes in myocardial strain were analyzed as independent variables. All 10 patients enrolled received a cumulative dose of doxorubicin of 300 mg/m. A comparison of pretreatment and posttreatment cMRI demonstrated 5 (50%) patients with a ≥10% decrease in LVEF (median, -8.4%; range, 1% to -17%; P=0.004). Three patients had at least 1 new or progressive segment of GD-DE. The global circumferential strain was significantly lower in patients after treatment, as compared with values before treatment (P=0.018) and to normal controls (P=0.046). Patients after treatment also had significantly lower global longitudinal strain than controls (P=0.035), and longitudinal strain values that tended to decrease compared with pretreatment values (P=0.073). Our data suggests that cMRI has the ability to assess both early structural and functional myocardial changes in association with doxorubicin-based chemotherapy.


Dorsal onlay graft urethroplasty using penile skin or buccal mucosa in adult bulbourethral strictures.
Preputial skin graft is used routinely for urethral reconstruction in patients with stricture disease. Alternative donor sites include extrapenile skin, bladder mucosa and buccal mucosa. Recently buccal mucosa graft has been suggested when local epithelial tissue is not available. We describe our experience with 37 patients undergoing 1-stage correction of bulbar urethral stricture using a penile skin (31) or buccal mucosa (6) graft. In 37 patients with bulbar urethral strictures a nontubularized dorsal onlay graft was used for urethral reconstruction. A preputial skin graft was used in 31 patients and a buccal mucosa graft in 6 with a paucity of local skin. Buccal mucosa graft length ranged from 2.5 to 5 cm. (average 4) and preputial skin graft was 2.5 to 12 cm. long (average 4.7). A dorsal approach to the urethral lumen was used in all patients who underwent onlay graft urethroplasty. Mean followup was 21.5 months for all 37 patients, 23 months for 31 treated with preputial skin graft and 13.5 months for 6 treated with buccal mucosa graft. The clinical outcomes were considered a failure anytime postoperative instrumentation was needed, including dilatation. In the series 34 cases (92%) were classified as a success and 3 (8%) as failure. Onlay graft urethroplasty provided excellent results in 92% of adults with bulbourethral stricture. The dorsal approach to the urethra allowed the use of foreskin or buccal mucosa graft for reconstruction of the adequate urethral lumen.


Mammary Analog Secretory Carcinoma (MASC) Involving the Thyroid Gland: A Report of the First 3 Cases.
Salivary gland-type tumors have been rarely described in the thyroid gland. Mammary Analog Secretory Carcinoma (MASC) is a recently defined type of salivary gland carcinoma characterized by a t(12;15)(p13;q25) resulting in an ETV6-NTRK3 fusion gene. We report 3 cases of MASC involving the thyroid gland without clinical evidence of a salivary gland or breast primary; the clinico-pathologic characteristics are reviewed. Assessment for rearrangement of the ETV6 (12p13) locus was conducted by fluorescence in situ hybridization (FISH) on representative FFPE sections using an ETV6 break apart probe (Abbott Molecular, Des Plaines, IL, USA). The patients were two females (52 and 55 years-old) and 1 male (74 years-old). The tumors were poorly circumscribed solid white tan nodules involving the thyroid. Histologically, they were invasive and showed solid, microcystic, cribriform, and tubular growth patterns composed of variably bland polygonal eosinophilic cells with vesicular nuclear chromatin and conspicuous nucleoli. All three cases showed metastasis to lymph nodes; one case showed lateral neck involvement. The tumor cells were positive for S100 and mammaglobin. GATA-3 and PAX-8 were positive in 2 cases, one of which only focally so. All three cases were negative for TTF-1 and thyroglobulin. Rearrangement of the ETV6 locus was confirmed in all cases and a diagnosis of MASC rendered for each case. A site of origin distinct from the thyroid gland was not identified, with a median follow up of 24 months. MASC may rarely involve the thyroid gland. The origin of these lesions is unknown; while an origin from ectopic salivary gland-type cells is entertained, a metastatic origin from an occult primary cannot be definitively excluded at this time. Given the histologic (follicular-like microcystic pattern with colloid-like secretions and papillary pattern), immunophenotypic (PAX-8), and even molecular overlap, MASC can be mistaken for papillary thyroid carcinoma and should be considered in the differential diagnosis of a thyroid mass.


The inlAB locus mediates the entry of Listeria monocytogenes into hepatocytes in vivo.
The intracellular parasite Listeria monocytogenes is able to induce its internalization by cultured mammalian cells that are not normally phagocytic. This process requires the expression of the chromosomal locus inlAB. We studied the virulence of an inlAB mutant and of its parent in murine listeriosis. Irrespective of the route of inoculation, the inlAB mutant was severely attenuated for growth in the liver. The livers of mice inoculated with the inlAB mutant displayed much smaller infectious foci than the parent as early as 24 h after infection. Electron microscopy showed that these foci consisted of a few inflammatory cells, with few bacteria; bacteria were rarely found within hepatocytes. In contrast, foci in livers of mice inoculated with the parent consisted of islets of heavily infected hepatocytes that were infiltrated by numerous neutrophils; bacteria seemed intact within hepatocytes and damaged within neutrophils. A direct role of inlAB for the entry of L. monocytogenes into hepatocytes was confirmed in a cell infection system using the murine embryonic hepatocyte cell line TIB73. The inlAB mutant was approximately 20-fold less invasive in trans. The "invasion locus" inlAB contributes to protect L. monocytogenes from the host's innate defense mechanisms by promoting its entry into hepatocytes.


New findings for maternal mortality age patterns: aggregated results for 38 countries.
With recent results showing a global decline in overall maternal mortality during the last two decades and with the target date for achieving the Millennium Development Goals only four years away, the question of how to continue or even accelerate the decline has become more pressing. By knowing where the risk is highest as well as where the numbers of deaths are greatest, it may be possible to re-direct resources and fine-tune strategies for greater effectiveness in efforts to reduce maternal mortality. We aggregate data from 38 Demographic and Health Surveys that included a maternal mortality module and were conducted in 2000 or later to produce maternal mortality ratios, rates, and numbers of deaths by five year age groups, separately by residence, region, and overall mortality level. The age pattern of maternal mortality is broadly similar across regions, type of place of residence, and overall level of maternal mortality. A "J" shaped curve, with markedly higher risk after age 30, is evident in all groups. We find that the excess risk among adolescents is of a much lower magnitude than is generally assumed. The oldest age groups appear to be especially resistant to change. We also find evidence of extremely elevated risk among older mothers in countries with high levels of HIV prevalence. The largest number of deaths occurs in the age groups from 20-34, largely because those are the ages at which women are most likely to give birth so efforts directed at this group would most effectively reduce the number of deaths. Yet equity considerations suggest that efforts also be directed toward those most at risk, i.e., older women and adolescents. Because women are at risk each time they become pregnant, fulfilling the substantial unmet need for contraception is a cross-cutting strategy that can address both effectiveness and equity concerns.


Forces between hydrophobic solids in concentrated aqueous salt solution.
Much research has focused on the discovery and description of long-ranged forces between hydrophobic solids immersed in water. Here we show that the force between high contact-angle solids in concentrated salt solution (1 M KCl) agrees very well with van der Waals forces calculated from Lifshitz theory for separations greater than 5 nm. The hydrophobic solids are octadecyltrichlorosilane-coated glass, with an advancing contact angle of 108°. Thus, in 1 M salt solution, it is unnecessary to invoke the presence of a hydrophobic force at separations greater than 5 nm. Through measurement in salt solution, we avoid the necessity of accounting for large electrostatic forces that frequently occur in pure water and may obscure resolution of other forces.


The significance of tumor persistence after incomplete excision of basal cell carcinoma.
Physicians inevitably receive a pathology report after excision of a basal cell carcinoma that indicates that it is incompletely excised. The physician and patient are then left with the dilemma of whether immediate re-excision or close clinical follow-up is indicated. Our purpose was to identify characteristics of incompletely excised basal cell carcinomas that are at low risk for recurrence. We retrospectively reviewed the charts and pathology slides of all incompletely excised basal cell carcinomas from 1991 to 1994 in a university hospital tumor registry. Incompletely excised basal cell carcinomas of superficial or nodular subtype, less than 1 cm in diameter, located anywhere except the nose or ears, with less than 4% marginal involvement on the initial inadequate excision had no evidence of tumor persistence. When physicians receive a pathology report indicating the incomplete excision of a basal cell carcinoma, they face the dilemma of further management. The majority of patients should undergo immediate re-excision or Mohs micrographic surgery because tumor persistence was found in 28% of cases. Occasionally, for a small group of select patients, close clinical follow-up may be indicated if the risk of recurrence is very low.


Effects of physical training on cardiac myosin ATPase activity.
Cardiac myosin from rats exercised 90 or 150 min daily for 8 wk was compared with the myosin from the hearts of matched sedentary controls. The Ca++-ATPase activity was increased 17 percent in rats exercised 90 min and 30 percent in rats exercised 150 min daily. In the exercised group 0.18 M KCl increased the myosin ATPase activity by 50 percent but had no effect in the control group. Ethylene glycol activated the Ca++-ATPase in control myosin preparations, but had no significant effect on myosin from conditioned hearts. Heavy meromyosin (HMM) from conditioned hearts had a higher Ca++-ATPase activity than from controls. Fluorescence with 8-anilinonaphthalene sulfonate (ANS) was increased 30 percent in HMM from conditioned hearts. The results suggest that the increased myosin ATPase activity in the hearts of exercised animals may be due to a local conformational change at or near the active site.


STED microscopy and its applications: new insights into cellular processes on the nanoscale.
For about a decade, superresolution fluorescence microscopy has been advancing steadily, maturing from the proof-of-principle stage to routine application. Of the various techniques, STED (stimulated emission depletion) microscopy was the first to break the diffraction barrier. Today, it is a prominent and versatile form of superresolution light microscopy. STED microscopy has shed a sharper light on numerous topics in cell biology, but also in material sciences. Both disciplines extend into the nanometer range, making detailed studies of structural and functional relationships difficult or even impossible to achieve using diffraction-limited microscopy. With recent advancements like spectral multiplexing or live-cell imaging, STED microscopy makes nanoscale materials and components of the cell accessible for fluorescence-based investigations. With multicolor superresolution imaging, even the interactions between biological and engineered nanostructures can be studied in detail. This review gives an introduction into the working principle of STED microscopy, provides a detailed overview of recent advancements and new techniques implemented for use with STED microscopy and shows how these have been applied in the life sciences and nanotechnologies.


TIS21 (/BTG2/PC3) as a link between ageing and cancer: cell cycle regulator and endogenous cell death molecule.
TIS21(/BTG2/PC3), orthologs of mouse, human and rat, respectively, is initially identified as one of the early growth response genes and induced by various stimulations. TIS21 belongs to antiproliferative (APRO) gene family containing the BTG-Box A (Y(50)-N(71)) and BTG-Box B (L(97)-E(115)), which are highly conserved among various species. On the other hand, it has lately been found that the expression of TIS21 is constitutive and high in thymus, lung alveolar epithelium, proximal tubule of kidney and basal cell layer of prostate acini. Potential roles of TIS21 have been suggested as transcriptional co-regulator, differentiation and antiapoptotic factor in neurogenesis, key mediator of the stage-specific expansion of thymocyte and negative regulator of hematopoietic progenitor expansion, and tumor suppressor gene in both mouse and human. In addition, as pan-cell cycle regulator TIS21 induces G1/S arrest by pRB dependently and pRB independently and G2/M arrest and cell death in the p53 null tumor cells, and regulates the development of vertebrate patterning in mouse, paraxial mesoderm development in zebrafish, and notochord development in Xenopus. It has been known that the expression of TIS21 depends on the induction of wt p53 when cells are damaged, however, it can also be upregulated p53 independently by the activation of PKC-delta pathway in tumor cells. The characteristic roles of TIS21 are discussed in the present review: (1) TIS21 inhibits early phase of carcinogenesis in its high expressers such as kidney, prostate, breast and thymus: Loss of constitutive and high expression of TIS21 was observed in the precancerous lesions as well as tumor tissues. As an endogenous cell death molecule, TIS21 may be involved in translocation of Pin-1 to cytoplasm. Pin-1 subsequently interacts with Serine(147) residue in TIS21 protein, resulting in mitochondrial depolarization. (2) TIS21 regulates transition of cell cycle at G1/S and G2/M phases in cancer cells with inactive pRB and/or p53, as well as in normal cells by regulating pRB/p16(INK4a) pathway. The latter has already been well elucidated; TIS21 inhibits the expression of cyclin D1, thus resulting in the arrest of cells at G1/S phase by pRB and p53 dependent manner. On the other hand, TIS21 inhibits degradations of cyclin A and cyclin B1 at G2/M phase, and directly binds to Cdc2, resulting in the failure of mitotic exit and then increasing the tumor cell death, when stimulated by high concentration of EGF. Therefore, TIS21 can be suggested as a pan-cell cycle modulator. (3) TIS21 regulates embryo development by activating BMP signal through interaction with Smad 1 and Smad 8, thereby regulating vertebral patterning in mice. It is also involved in notochord development in Xenopus and paraxial mesoderm development in zebrafish. Based on the previous report that the expression of TIS21 is involved in the induction of senescence after chemotherapy of cancer cells, which can be a mechanism to resist carcinogenesis, TIS21(/BTG2/PC3), the endogenous cell death molecule and pan-cell cycle regulator, might be a link between cellular senescence and carcinogenesis.


The Association Between Condomless Anal Sex and Social Support Among Black Men Who Have Sex With Men (MSM) in Six U.S. Cities: A Study Using Data from the HIV Prevention Trials Network BROTHERS Study (HPTN 061).
We assessed how egocentric (i.e., self-generated descriptions of a person's social contacts) network structure and composition corresponded with reported instances of condomless receptive and insertive anal intercourse with men who were reportedly HIV-infected or of unknown HIV serostatus in a sample of black men who have sex with men (MSM) in six U.S. cities. Ratings showing a higher percentage of network members who provided social participation and medical support were positively associated with reporting condomless sex. There were also significant positive associations between stimulant use and condomless insertive and receptive anal sex. Future research should examine the social processes that underlie these associations and explore ways that social support can affect HIV prevention efforts for black MSM.


Neonatal gram-negative infections, antibiotic susceptibility and clinical outcome: an observational study.
Neonatal gram-negative (GN) infections are associated with high mortality and morbidity. Early appropriate antibiotic treatment is vital and gentamicin is the most frequently used antibiotic on neonatal units (NNUs). Antimicrobial breakpoints are predominantly based on adult data and the relationship between minimum inhibitory concentrations (MICs) and outcome in neonates is unclear. We aimed to determine the MIC of GN pathogens causing neonatal infections and relate this to clinical outcomes. MICs for eight antibiotics plus extended spectrum β-lactamase (ESBL) production were determined for invasive GN bacterial isolates from eight UK NNUs. European Committee on Antimicrobial Susceptibility Testing breakpoints were applied. MIC was correlated with clinical outcome using multivariable regression analysis. 118 isolates from 116 patients were analysed. The median birth gestation and postnatal age was 27 weeks (IQR 24.6-32.3) and 20 days (IQR 5-44), respectively. Pathogens included Escherichia coli (51%), Klebsiella spp (23%) and Enterobacter spp (22%). 10-day attributable mortality was 18.1% (21 patients) with the highest mortality from Pseudomonas aeruginosa infections. ESBL producers accounted for 13.8% of the isolates. In regression analysis, increasing gentamicin MIC was associated with increased mortality in gentamicin treated patients across the full MIC range (OR per loge increase in MIC: 2.29; 95% CI 1.23 to 4.26, p=0.009), including susceptible isolates only (MIC ≤4) (OR 3.05; 95% CI 1.10 to 8.46, p=0.032). Neonatal mortality from GN infections remains high and is associated with increasing gentamicin MIC, even for isolates deemed susceptible. A better understanding of population-specific MICs and aminoglycoside dosing is required to guide empiric antibiotic treatment.


Dual-source versus 64-section CT coronary angiography at lower heart rates: comparison of accuracy and radiation dose.
To compare the diagnostic performance and radiation doses of dual-source and 64-section computed tomographic (CT) coronary angiography for the diagnosis of significant coronary stenoses in patients with heart rates of 65 beats/min or less. This retrospective study had local ethics committee approval; all patients gave written informed consent. Two hundred patients with heart rates of 65 beats/min or less were enrolled; 100 underwent dual-source and 100 underwent 64-section CT coronary angiography. Two blinded observers independently assessed image quality of all coronary segments by using a four-point scale and searched for significant (>50%) stenoses in each segment. Catheter angiography was used as the reference standard. Image noise was measured in the ascending aorta. Radiation doses were calculated. No significant differences were found regarding sex, age, body weight, cardiovascular risk profile, prevalence of stenosis, mean and variability of heart rate, Agatston score, and image noise (all P > .1) between patients in both CT coronary angiography groups. No significant difference was found in the rate of nonassessable coronary segments between dual-source (1.0%, 14 of 1405) and 64-section CT coronary angiography (1.8%, 25 of 1387; P = .08). Motion artifacts occurred significantly more often in 64-section (21 of 25) versus dual-source (five of 14, P = .004) CT coronary angiography. Segment-based accuracy and specificity were significantly higher for dual-source versus 64-section CT coronary angiography. There was no significant difference in accuracy parameters at the per-vessel and per-patient analyses. No significant difference (P = .13) was found between the effective doses of dual-source (mean +/- standard deviation, 10.9 mSv +/- 1.1) and 64-section CT (10.4 mSv +/- 1.7) coronary angiography. In patients with heart rates of 65 beats/min or less, the higher temporal resolution of dual-source CT coronary angiography results in improved accuracy and specificity for the diagnosis of significant stenoses on a per-segment level at a similar radiation dose, but provides a comparable diagnostic accuracy on a patient-based level as does 64-section coronary angiography.


Humidity-enhanced sub-ppm sensitivity to ammonia of covalently functionalized single-wall carbon nanotube bundle layers.
A low-cost method for carbon nanotubes (CNTs) network production from solutions on flexible polyethylene naphthalate substrates has been adopted to prepare high quality and well characterized SWCNT bundle layers to be used as the active layer in chemiresistor gas sensors. Two types of SWCNTs have been tested: pristine SWCNTs, deposited from a surfactant solution, and covalently functionalized SWCNTs, deposited from a dimethyl-acetamide solution. The humidity effects on the sensitivity of the SWCNTs network to NH3 have been investigated. The results show that relative humidity favors the response to NH3, confirming recent theoretical predictions. The COOH-functionalized sample displays the largest response owing to both its hydrophilic nature, favoring the interaction with H2O molecules, and its largest surface area. Compared to data available in the literature, the present sensors display a remarkable sensitivity well below the ppm range, which makes them quite promising for environmental and medical applications, where NH3 concentrations (mostly of the order of tens of ppb) have to be detected.


Identification and quantitation of unsaturated fatty acid isomers by electrospray ionization tandem mass spectrometry: a shotgun lipidomics approach.
Identification and quantification of unsaturated fatty acid (FA) isomers in a biological system are significant in the study of lipid metabolism and catabolism, membrane biophysics, and pathogenesis of diseases but are challenging in lipidomics. We developed a novel approach for identification and quantitation of unsaturated FA isomers by exploiting two facts: (1) unsaturated FA anions yield fragment ion(s) from loss of CO(2) or H(2)O from the anions upon collision-induced dissociation; and (2) the fragment ions yielded from discrete FA isomers have distinct profiles of the fragment ion intensity vs. collision conditions. These distinct profiles likely result from the differential interactions of the negative charge of the fragment ion with the electron clouds of the double bonds due to their different distances in discrete FA isomers. The novel approach was also extended to analyze the double bond isomers of FA chains present in phospholipids by multistage tandem mass spectrometry. Collectively, we developed a new approach for identification and quantification of the double bond isomers of endogenous FA species or FA chains present in intact phospholipid species. We believe that this approach should further advance the lipidomic power for identification of the biochemical mechanisms underlying metabolic diseases.


Change of properties during storage of a UDMA/TEGDMA dental resin.
The aim of this study was to evaluate the changes in viscoelastic properties of a UDMA-based dental resin as a function of time after initial light exposure. Specimens of a UDMA/TEGDMA (70:30 wt%) resin were irradiated by a visible-light-curing unit. Immediately after the irradiation, the light-cured specimen was stored in the dark for different times from 1 to 120 h at 37 degrees C, and characterized by means of DMA, DSC, and FTIR spectroscopy. The irradiated specimen exhibited a bimodal shape in the form of two rapid declines in log E' corresponding to glass transition with a plateau between the two declines. Two distinct peaks were seen in tan delta versus temperature. The thermal reaction of the incompletely cured sample with residual groups trapped by the fast reaction during irradiation is responsible for the plateau. After storage, significant changes were observed in dynamic mechanical parameters, DSC exotherm, and degree of conversion. Storage modulus continued to increase during the 4 h of storage and leveled off thereafter. Peak heights of tan delta versus temperature were also influenced by storage. Degree of conversion increased from 75 +/- 2% immediately after irradiation to 87 +/- 3% after 120 h storage. The changes of the properties of this dental resin system when stored at 37 degrees C after irradiation are clinically important in terms of stability, durability, and performance after initial polymerization.


Activation of heat shock and antioxidant responses by the natural product celastrol: transcriptional signatures of a thiol-targeted molecule.
Stress response pathways allow cells to sense and respond to environmental changes and adverse pathophysiological states. Pharmacological modulation of cellular stress pathways has implications in the treatment of human diseases, including neurodegenerative disorders, cardiovascular disease, and cancer. The quinone methide triterpene celastrol, derived from a traditional Chinese medicinal herb, has numerous pharmacological properties, and it is a potent activator of the mammalian heat shock transcription factor HSF1. However, its mode of action and spectrum of cellular targets are poorly understood. We show here that celastrol activates Hsf1 in Saccharomyces cerevisiae at a similar effective concentration seen in mammalian cells. Transcriptional profiling revealed that celastrol treatment induces a battery of oxidant defense genes in addition to heat shock genes. Celastrol activated the yeast Yap1 oxidant defense transcription factor via the carboxy-terminal redox center that responds to electrophilic compounds. Antioxidant response genes were likewise induced in mammalian cells, demonstrating that the activation of two major cell stress pathways by celastrol is conserved. We report that celastrol's biological effects, including inhibition of glucocorticoid receptor activity, can be blocked by the addition of excess free thiol, suggesting a chemical mechanism for biological activity based on modification of key reactive thiols by this natural product.


Control of macrophage Ia expression in neonatal mice--role of a splenic suppressor cell.
The control of macrophage expression of I region-associated antigens (Ia) in neonatal mice was studied by comparing responses of neonatal and adult mice to immune vs nonimmune stimuli. Adults generated peritoneal exudates rich in Ia-bearing macrophages in response to i.p. injection of live Listeria monocytogenes, Listeria-immune T cells, and heat-killed Listeria, or a soluble mediator termed macrophage Ia-recruiting factor (MIRF). Neonates failed to respond to these stimuli. In contrast, both neonates and adults generated Ia-negative peritoneal exudates when stimulated with thioglycollate. A neonatal spleen cell that blocked the response of adults both to immune T cells and heat-killed Listeria and to MIRF was identified and characterized. Some of the suppressor cells appeared to be early precursors of the phagocytic lineage that develop into mature monocyte-macrophages. Suppression was apparently mediated by metabolites of arachidonic acid since indomethacin and aspirin in vivo blocked the effect. Similar suppressor activity was found in adult bone marrow and in adult resident peritoneal exudate cells. Thus, the phagocytic line autoregulates its surface expression of Ia in both neonatal and adult mice. This mechanism becomes particularly pointed during early development and could contribute to the lack of immunity during ontogeny.


Interventions to improve medication reconciliation in primary care.
To systematically review all primary care intervention studies designed to implement medication reconciliation for effects on medication discrepancies, clinical outcomes, and patient knowledge of their medications. We searched MEDLINE (1988-March 2008); Healthstar (1966-March 2008); CINAHL (1982-March 2008); EMBASE (1980-March 2008); Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, Database of Abstracts of Reviews of Effects, Cochrane Methodology Register, and Health Technology Assessments; and unpublished material. No language restrictions were applied. Search terms included medication reconciliation, medication errors, prescribing error, medication systems, adverse drug events, drug utilization review, medication list, medication record, and medications management. Randomized controlled trials or before-and-after studies that examined the effect of various interventions on medication discrepancies either in ambulatory settings or at hospital discharge among community-dwelling adults were included. Two reviewers independently assessed studies to determine inclusion. Level of agreement between the reviewers was good, with unweighted Cohen's kappa of 0.71. Two of 3 independent reviewers abstracted data and evaluated validity from included studies. Disagreements between reviewers were resolved by consensus. Four trials met the inclusion criteria. Two before-and-after studies (n = 275) in ambulatory care examining systematic medication reconciliation at each visit produced conflicting results. One study showed a reduction in the proportion of medication discrepancies from 88.5% to 49.1% (OR 0.13; 95% CI 0.07 to 0.21); the other showed no benefit. One randomized controlled trial and one before-and-after study (n = 202) evaluated pharmacist medication review at hospital discharge. Neither showed a benefit. Heterogeneity precluded pooling of studies. All included studies had significant design flaws. There is no good quality evidence demonstrating the effectiveness of medication reconciliation in the primary care setting. Further research is needed.


Purification and characterization of human cell--cell adhesion molecule 1 (C-CAM1) expressed in insect cells.
The cell--cell adhesion molecule 1 (C-CAM1) plays an important role as a tumor suppressor for prostate cancer. Decreased expression of C-CAM1 was detected in prostate, breast, and colon carcinoma. Reexpression of C-CAM1 in prostate and breast cancer cell lines was able to suppress tumorigenicity in vivo. These observations suggest that C-CAM1 may be used as a marker for cancer detection or diagnosis. To generate monoclonal antibodies specific to C-CAM1, we have overexpressed full-length human C-CAM1 in Sf9 cells using a baculovirus expression system. The protein was purified 104-fold using nickel affinity chromatography. About 0.4 mg purified C-CAM1 was obtained from 200 mg of infected cells. When the purified protein was digested with peptidyl-N-glycosidase, the apparent mobility of the protein on SDS--PAGE changed from 90 to 58 kDa, which is close to the molecular weight predicted from the cloned cDNA sequence. This observation suggests that C-CAM1 was glycosylated on asparagine residues when expressed in Sf9 cells. Western blotting and internal protein sequencing analysis confirmed that the purified protein is human C-CAM1. Biochemical and functional assays indicate that this protein expressed in Sf9 cells displays characteristics similar to those of native protein, including adhesion function and glycosylation modification. Using this protocol, sufficient quantity of this protein can be produced with purity suitable for monoclonal antibody generation and biochemical study.


A mycobacterium ESX-1-secreted virulence factor with unique requirements for export.
Specialized secretion systems of pathogenic bacteria commonly transport multiple effectors that act in concert to control and exploit the host cell as a replication-permissive niche. Both the Mycobacterium marinum and the Mycobacterium tuberculosis genomes contain an extended region of difference 1 (extRD1) locus that encodes one such pathway, the early secretory antigenic target 6 (ESAT-6) system 1 (ESX-1) secretion apparatus. ESX-1 is required for virulence and for secretion of the proteins ESAT-6, culture filtrate protein 10 (CFP-10), and EspA. Here, we show that both Rv3881c and its M. marinum homolog, Mh3881c, are secreted proteins, and disruption of RD1 in either organism blocks secretion. We have renamed the Rv3881c/Mh3881c gene espB for ESX-1 substrate protein B. Secretion of M. marinum EspB (EspBM) requires both the Mh3879c and Mh3871 genes within RD1, while CFP-10 secretion is not affected by disruption of Mh3879c. In contrast, disruption of Mh3866 or Mh3867 within the extRD1 locus prevents CFP-10 secretion without effect on EspBM. Mutants that fail to secrete only EspBM or only CFP-10 are less attenuated in macrophages than mutants failing to secrete both substrates. EspBM physically interacts with Mh3879c; the M. tuberculosis homolog, EspBT, physically interacts with Rv3879c; and mutants of EspBM that fail to bind Mh3879c fail to be secreted. We also found interaction between Rv3879c and Rv3871, a component of the ESX-1 machine, suggesting a mechanism for the secretion of EspB. The results establish EspB as a substrate of ESX-1 that is required for virulence and growth in macrophages and suggests that the contribution of ESX-1 to virulence may arise from the secretion of multiple independent substrates.


High strength binding of P815 mastocytoma cells is not necessary for their lysis by macrophages which have been primed and triggered in vitro.
We have examined the hypothesis that binding of P815 mastocytoma cells is a necessary step in lysis of these cells by macrophages which are both "primed" and "triggered" in vitro, Macrophages "primed" by conditioned media containing IFN-gamma, or by rIFN-gamma have an increased ability to bind P815. However, adding either heat-killed Listeria or endotoxin to trigger the primed macrophages has opposite effects on lysis and binding of P815. Lysis is increased. Binding is dramatically decreased. This is true when centrifugal forces of 200 x g, 400 x g, and 800 x g are used to disrupt P815-macrophage binding. Although 100% of P815 cells bound by cytotoxic macrophages are lysed, a large additional population of unbound P815 is also lysed. Detailed kinetic studies indicate that macrophages do not rapidly bind and lyse several cycles of P815. There is an initial lag period of 4 to 6 h before P815 lysis can be detected, and completion of lytic events then occurs within 12 to 14 h. Lysis of P815 bound to cytotoxic macrophages is slightly slower than lysis of the total population of bound and unbound P815. In contrast, D3.1, a cloned CD4+ T cell line, is tightly bound to macrophages but not lysed efficiently. When macrophages are simultaneously confronted with P815 and macrophage-bound D3.1, only the former are lysed. Altogether, the data indicate that P815-macrophage binding, as operationally defined by our assay, is not a necessary step for lysis. These results, by use of macrophages primed and triggered in vitro, are in contrast to previously reported experiments examining P815 binding and lysis by macrophages activated in vivo by infection with bacillus Calmette-Guérin.


Cdc42 controls progenitor cell differentiation and beta-catenin turnover in skin.
Differentiation of skin stem cells into hair follicles (HFs) requires the inhibition of beta-catenin degradation, which is controlled by a complex containing axin and the protein kinase GSK3beta. Using conditional gene targeting in mice, we show now that the small GTPase Cdc42 is crucial for differentiation of skin progenitor cells into HF lineage and that it regulates the turnover of beta-catenin. In the absence of Cdc42, degradation of beta-catenin was increased corresponding to a decreased phosphorylation of GSK3beta at Ser 9 and an increased phosphorylation of axin, which is known to be required for binding of beta-catenin to the degradation machinery. Cdc42-mediated regulation of beta-catenin turnover was completely dependent on PKCzeta, which associated with Cdc42, Par6, and Par3. These data suggest that Cdc42 regulation of beta-catenin turnover is important for terminal differentiation of HF progenitor cells in vivo.


UNC-16/JIP3 regulates early events in synaptic vesicle protein trafficking via LRK-1/LRRK2 and AP complexes.
JIP3/UNC-16/dSYD is a MAPK-scaffolding protein with roles in protein trafficking. We show that it is present on the Golgi and is necessary for the polarized distribution of synaptic vesicle proteins (SVPs) and dendritic proteins in neurons. UNC-16 excludes Golgi enzymes from SVP transport carriers and facilitates inclusion of specific SVPs into the same transport carrier. The SVP trafficking roles of UNC-16 are mediated through LRK-1, whose localization to the Golgi is reduced in unc-16 animals. UNC-16, through LRK-1, also enables Golgi-localization of the μ-subunit of the AP-1 complex. AP1 regulates the size but not the composition of SVP transport carriers. Additionally, UNC-16 and LRK-1 through the AP-3 complex regulates the composition but not the size of the SVP transport carrier. These early biogenesis steps are essential for dependence on the synaptic vesicle motor, UNC-104 for axonal transport. Our results show that UNC-16 and its downstream effectors, LRK-1 and the AP complexes function at the Golgi and/or post-Golgi compartments to control early steps of SV biogenesis. The UNC-16 dependent steps of exclusion, inclusion and motor recruitment are critical for polarized distribution of neuronal cargo.


Ribozyme-mediated trans-splicing of a trinucleotide repeat.
Trinucleotide repeat expansions (TREs) are a recently described class of mutations characterized by a change in the size of the genomic fragment due to amplification of the repeated unit. A number of diseases have been attributed to TRE, including Huntington disease and myotonic dystrophy (DM), but attempts at genetic therapy have yet to prove successful. A potential therapeutic approach would be to repair the expanded repeat using the trans-splicing ability of group I intron ribozymes. We have used DM as a model to test this hypothesis. A group I intron ribozyme (DMPK-RZ1) was designed to modify the TRE at the 3' end of the human myotonic dystrophy protein kinase (DMPK) transcripts. DMPK-RZ1 was shown to ligate a small DMPK mRNA fragment, contained within the ribozyme, to a simple DMPK-target RNA in vitro. It also modified a larger target transcript, leading to replacement of twelve repeats with five repeats, both in vitro and in mammalian cells. Finally, this ribozyme successfully replaced the 3' end of endogenous DMPK mRNA in fibroblasts with a different 3' region. Ribozyme-mediated RNA repair may thus form a novel therapeutic strategy for diseases associated with repeat expansions.


Interaction of Escherichia coli with growing salad spinach plants.
In this study, the interaction of a bioluminescence-labeled Escherichia coli strain with growing spinach plants was assessed. Through bioluminescence profiles, the direct visualization of E. coli growing around the roots of developing seedlings was accomplished. Subsequent in situ glucuronidase (GUS) staining of seedlings confirmed that E. coli had become internalized within root tissue and, to a limited extent, within hypocotyls. When inoculated seeds were sown in soil microcosms and cultivated for 42 days, E. coli was recovered from the external surfaces of spinach roots and leaves as well as from surface-sterilized roots. When 20-day-old spinach seedlings (from uninoculated seeds) were transferred to soil inoculated with E. coli, the bacterium became established on the plant surface, but internalization into the inner root tissue was restricted. However, for seedlings transferred to a hydroponic system containing 10(2) or 10(3) CFU of E. coli per ml of the circulating nutrient solution, the bacterium was recovered from surface-sterilized roots, indicating that it had been internalized. Differences between E. coli interactions in the soil and those in the hydroponic system may be attributed to greater accessibility of the roots in the latter model. Alternatively, the presence of a competitive microflora in soil may have restricted root colonization by E. coli. The implications of this study's findings with regard to the microbiological safety of minimally processed vegetables are discussed.


Mast cell tryptase and proteinase-activated receptor 2 induce hyperexcitability of guinea-pig submucosal neurons.
Mast cells that are in close proximity to autonomic and enteric nerves release several mediators that cause neuronal hyperexcitability. This study examined whether mast cell tryptase evokes acute and long-term hyperexcitability in submucosal neurons from the guinea-pig ileum by activating proteinase-activated receptor 2 (PAR2) on these neurons. We detected the expression of PAR2 in the submucosal plexus using RT-PCR. Most submucosal neurons displayed PAR2 immunoreactivity, including those colocalizing VIP. Brief (minutes) application of selective PAR2 agonists, including trypsin, the activating peptide SL-NH2 and mast cell tryptase, evoked depolarizations of the submucosal neurons, as measured with intracellular recording techniques. The membrane potential returned to resting values following washout of agonists, but most neurons were hyperexcitable for the duration of recordings (> 30 min-hours) and exhibited an increased input resistance and amplitude of fast EPSPs. Trypsin, in the presence of soybean trypsin inhibitor, and the reverse sequence of the activating peptide (LR-NH2) had no effect on neuronal membrane potential or long-term excitability. Degranulation of mast cells in the presence of antagonists of established excitatory mast cell mediators (histamine, 5-HT, prostaglandins) also caused depolarization, and following washout of antigen, long-term excitation was observed. Mast cell degranulation resulted in the release of proteases, which desensitized neurons to other agonists of PAR2. Our results suggest that proteases from degranulated mast cells cleave PAR2 on submucosal neurons to cause acute and long-term hyperexcitability. This signalling pathway between immune cells and neurons is a previously unrecognized mechanism that could contribute to chronic alterations in visceral function.


Electron microscopy of extracellular materials during the development of a sea star, Patiria miniata (Echinodermata: Asteroidea).
The fine structure of conspicuous extracellular materials during the life history of a sea star (Patiria miniata) is described. The outer surface of the developing sea star is covered by two morphologically different cuticles that appear sequentially during ontogeny. The primary cuticle, which is about 120 nm thick and two-layered, is present from mid-blastula through the end of the larval stage. The secondary cuticle, which is about 1 micron thick and three-layered, first appears on the epidermis of the rudiment region of the larva and, after metamorphosis, covers the entire epidermis of the juvenile and adult stages. During ontogeny, there are only two conspicuous gut cuticles: the first lines the newly invaginated archenteron at the start of the gastrula stage, and the second lines the esophagus during the larval stage. A blastocoelic basal lamina first appears at mid-blastula and persists as subectodermal and subendodermal basal laminae. Ruthenium red-positive granules are detectable between the lateral surfaces of adjacent ectodermal cells during part of the gastrula stage; this transient intercellular material may possibly aid in lateral adhesion between cells.


Carbohydrate-rich breakfast attenuates glycaemic, insulinaemic and ghrelin response to ad libitum lunch relative to morning fasting in lean adults.
Breakfast omission is associated with obesity and CVD/diabetes, but the acute effects of extended morning fasting upon subsequent energy intake and metabolic/hormonal responses have received less attention. In a randomised cross-over design, thirty-five lean men (n 14) and women (n 21) extended their overnight fast or ingested a typical carbohydrate-rich breakfast in quantities relative to RMR (i.e. 1963 (sd 238) kJ), before an ad libitum lunch 3 h later. Blood samples were obtained hourly throughout the day until 3 h post-lunch, with subjective appetite measures assessed. Lunch intake was greater following extended fasting (640 (sd 1042) kJ, P< 0.01) but incompletely compensated for the omitted breakfast, with total intake lower than the breakfast trial (3887 (sd 1326) v. 5213 (sd 1590) kJ, P< 0.001). Systemic concentrations of peptide tyrosine-tyrosine and leptin were greater during the afternoon following breakfast (both P< 0.05) but neither acylated/total ghrelin concentrations were suppressed by the ad libitum lunch in the breakfast trial, remaining greater than the morning fasting trial throughout the afternoon (all P< 0.05). Insulin concentrations were greater during the afternoon in the morning fasting trial (all P< 0.01). There were no differences between trials in subjective appetite during the afternoon. In conclusion, morning fasting caused incomplete energy compensation at an ad libitum lunch. Breakfast increased some anorectic hormones during the afternoon but paradoxically abolished ghrelin suppression by the second meal. Extending morning fasting until lunch altered subsequent metabolic and hormonal responses but without greater appetite during the afternoon. The present study clarifies the impact of acute breakfast omission and adds novel insights into second-meal metabolism.


Determination of alpha-bisabolol in human blood by micro-HPLC-ion trap MS and head space-GC-MS methods.
Alpha-bisabolol is a compound present in some essential oils, widely distributed in several plants, including camomile. Two different methods for analysing an essential oil, such as alpha-bisabolol in human blood are reported: the first uses micro-liquid chromatography-electrospray ionisation-mass spectrometry (muHPLC-ESI-MS), whereas the second is based on "head space" injection coupled to gas chromatography-mass spectrometry (HS-GC-MS). For LC-ESI-MS, human blood samples, spiked with alpha-bisabolol, were extracted with hexane and evaporated to dryness under air stream. The residue was then reconstituted with methanol and injected into a C18 column, connected to an ion trap mass spectrometer equipped with an ESI source. Spectra were recorded in the positive ion, selected ion monitoring mode. The detection limit of alpha-bisabolol in blood was 0.125 micromol/l. The preparation of samples for the analysis in HS-GC-MS was limited to blood dilution with water (0.5 ml blood + 1 ml water). Head space vials were heated at 125 degrees C for 1 h before automatic injection. The HS-GC-MS detection limit (0.13 micromol/l) was similar to that achieved with the muHPLC-ESI-MS method. Successful tests were performed to verify if alpha-bisabolol could be directly measured by the HS-GC-MS method in different biological samples (blood, urine, faeces, homogenate tissues) from rats treated with the camomile essential oil.


Xanthine oxidase mediates hypoxia-inducible factor-2α degradation by intermittent hypoxia.
Sleep-disordered breathing with recurrent apnea produces chronic intermittent hypoxia (IH). We previously reported that IH leads to down-regulation of HIF-2α protein via a calpain-dependent signaling pathway resulting in oxidative stress. In the present study, we delineated the signaling pathways associated with calpain-dependent HIF-2α degradation in cell cultures and rats subjected to chronic IH. Reactive oxygen species (ROS) scavengers prevented HIF-2α degradation by IH and ROS mimetic decreased HIF-2α protein levels in rat pheochromocytoma PC12 cell cultures, suggesting that ROS mediate IH-induced HIF-2α degradation. IH activated xanthine oxidase (XO) by increased proteolytic conversion of xanthine dehydrogenase to XO. ROS generated by XO activated calpains, which contributed to HIF-2α degradation by IH. Calpain-induced HIF-2α degradation involves C-terminus but not the N-terminus of the HIF-2α protein. Pharmacological blockade as well as genetic knock down of XO prevented IH induced calpain activation and HIF-2α degradation in PC12 cells. Systemic administration of allopurinol to rats prevented IH-induced hypertension, oxidative stress and XO activation in adrenal medulla. These results demonstrate that ROS generated by XO activation mediates IH-induced HIF-2α degradation via activation of calpains.


Cellular mechanism of the conduction abnormalities induced by serum from anti-Ro/SSA-positive patients in rabbit hearts.
In this study, IgG fractions from sera of SLE patients with anti-Ro/SSA or anti-Ro/SSA and anti-La/SSB activity were tested in Langendorff preparations of adult rabbit hearts, aiming to reproduce the cardiac manifestations observed in neonatal lupus in an experimental model. The hearts were perfused with normal Tyrode's solution for 30 min, followed by perfusion with Tyrode's containing 0.3 mg/ml of anti-Ro/SSA- (or anti-Ro/La-) positive IgG (nine sera), anti-ribonucleoprotein (RNP)-positive IgG (five sera), or IgG fractions from normal donors (five sera). In one third of the experiments done with anti-Ro/La-positive IgG, heart block was observed. With the remaining fractions, a decrease in heart rate of 17.1% was observed, but normal sinus rhythm was maintained. The IgG fractions with anti-RNP activity (five experiments) and from normal sera (six experiments) reduced heart rates by 12.9 and 3.3%, respectively, but heart block was not observed. To further characterize the cellular mechanisms involved in the conduction disturbances observed in the whole rabbit hearts, we conducted experiments with ventricular myocytes isolated from young rabbit hearts, studied by whole cell patch-clamp technique. In these experiments, the slow inward currents were analyzed during the superfusion of the cell with normal Tyrode's solution and 5 min after superfusion with Tyrode's solution containing 0.3 mg/ml of anti-Ro/SSA- (or anti-Ro/La-) positive IgG (five sera), anti-RNP-positive IgG (three sera), or IgG from normal donors (four sera). Resting and action potential amplitudes were not affected by any of the sera used. The anti-Ro/SSA IgG fraction induced a mean reduction in the peak slow inward current of 31.6%. IgG fractions with anti-RNP activity reduced slow inward current by 4.4%, whereas IgG fractions from normal donors increased this current by 3.3%. IgG-free fractions from sera of patients with anti-Ro/SSA activity did not alter the peak slow inward current. These results show, for the first time, that the presence of anti-Ro/SSA or anti-Ro/SSA and anti-La/SSB antibody activity in IgG fractions from lupus patients' sera can induce cardiac conduction disorders similar to those observed in neonatal lupus.


Pressure and flow properties of cannulae for extracorporeal membrane oxygenation I: return (arterial) cannulae.
Adequate extracorporeal membrane oxygenation support in the adult requires cannulae permitting blood flows up to 6-8 L/minute. In accordance with Poiseuille's law, flow is proportional to the fourth power of cannula inner diameter and inversely proportional to its length. Poiseuille's law can be applied to obtain the pressure drop of an incompressible, Newtonian fluid (such as water) flowing in a cylindrical tube. However, as blood is a pseudoplastic non-Newtonian fluid, the validity of Poiseuille's law is questionable for prediction of cannula properties in clinical practice. Pressure-flow charts with non-Newtonian fluids, such as blood, are typically not provided by the manufacturers. A standardized laboratory test of return (arterial) cannulae for extracorporeal membrane oxygenation was performed. The aim was to determine pressure-flow data with human whole blood in addition to manufacturers' water tests to facilitate an appropriate choice of cannula for the desired flow range. In total, 14 cannulae from three manufacturers were tested. Data concerning design, characteristics, and performance were graphically presented for each tested cannula. Measured blood flows were in most cases 3-21% lower than those provided by manufacturers. This was most pronounced in the narrow cannulae (15-17 Fr) where the reduction ranged from 27% to 40% at low flows and 5-15% in the upper flow range. These differences were less apparent with increasing cannula diameter. There was a marked disparity between manufacturers. Based on the measured results, testing of cannulae including whole blood flows in a standardized bench test would be recommended.


Maternal Psychological Distress and Executive Functions Are Associated During Early Parenthood - A FinnBrain Birth Cohort Study.
Parental executive functioning (EF) and parenting behaviors can be affected by the multiple stressors that are often present during early parenthood. However, little is known about how commonly experienced psychological distress during early parenthood is associated with parental EF capacity. We explored the links between psychological distress and EFs in a general population sample of 150 Finnish birth cohort mothers with 2.5-year-old children. The symptoms of depression, anxiety, insomnia, and poor couple relationship adjustment were measured with the self-report questionnaires EPDS, SCL-90, AIS, and RDAS. EFs were assessed with five computerized Cogstate tasks. When the psychological distress measures were added to a hierarchical regression analysis as continuous variables, no significant single or additive associations with EFs were found. When the distress measures were dichotomized to compare symptoms below/above cutoffs indicating clinically elevated levels, single distress domains remained as non-significant predictors, but a cumulative risk index of the number of concurrent clinically elevated distress domains was significantly associated with EFs. Thus, mothers with a higher number of concurrent clinically elevated psychological distress domains (i.e., depression, anxiety, insomnia, and poor couple relationship adjustment) tended to have lower EFs. This association is possibly bi-directional - clinically elevated distress within several domains could have a cumulative, depleting effect on maternal EF capacity, but a lower EF capacity could also increase the vulnerability for experienced distress within several concurrent domains. Longitudinal studies are needed to clarify potential causal links between stressors and EF.


Serum microRNA profiles among dioxin exposed veterans with monoclonal gammopathy of undetermined significance.
Previously an increased risk for monoclonal gammopathy of undetermined significance (MGUS), a precursor of multiple myeloma (MM), was reported among Vietnam veterans exposed to Agent Orange and its contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Dysregulated expression of certain microRNAs (miRNAs) was demonstrated in MGUS and MM. Given the important role of miRNAs in cellular homeostasis, the aim of this study was to determine if there was an association between serum levels of selected miRNAs and TCDD in 47 MGUS cases identified in our previous investigation using serum specimens and exposure data archived by the Air Force Health Study (AFHS). A total of 13 miRNA levels (let-7a, let-7i, miR-16, miR-20a, miR-21, miR-34a, miR-106b, miR-146a, miR-181a, miR-192, miR-205, miR-335, and miR-361) was measured in serum stored during the 2002 AFHS follow-up and the relationship to lipid-adjusted serum TCDD levels in 1987 was determined. miR-34a showed the strongest relationship with TCDD; after age-adjustment, this positive association was more pronounced. In contrast, the other 12 miRNAs displayed absolute values of age adjusted coefficient estimates below 1.16 and non-significant p-values. The observed strong positive association between high body burdens of TCDD and miR-34a, a tumor suppressor regulated by p53, in this MGUS population warrants clarification of the TCDD-miR-34a relationship and its role in the pathogenesis of MGUS and risk for MM.


Kernel methods for melanoma recognition.
Skin cancer is a spreading disease in the western world. Early detection and treatment are crucial for improving the patient survival rate. In this paper we present two algorithms for computer assisted diagnosis of melanomas. The first is the support vector machines algorithm, a state-of-the-art large margin classifier, which has shown remarkable performances on object recognition and categorization problems. The second method, spin glass-Markov random fields, combines results of statistical physics of spin glasses with Markov random fields. We compared the two approaches using color histograms as features. We benchmarked our methods with another algorithm presented in the literature, which uses a sophisticated segmentation technique and a set of features especially designed for melanoma recognition. To our knowledge, this algorithm represents the state of the art on skin lesions classification. We show with extensive experiments that the support vector machines approach outperforms the existing method and, on two classes out of three, it achieves performances comparable to those obtained by expert clinicians.


Adenylyl cyclase isoform 1 contributes to sinoatrial node automaticity via functional microdomains.
Sinoatrial node (SAN) cells are the heart's primary pacemaker. Their activity is tightly regulated by β-adrenergic receptor (β-AR) signaling. Adenylyl cyclase (AC) is a key enzyme in the β-AR pathway that catalyzes the production of cAMP. There are current gaps in our knowledge regarding the dominant AC isoforms and the specific roles of Ca2+-activated ACs in the SAN. The current study tests the hypothesis that distinct AC isoforms are preferentially expressed in the SAN and compartmentalize within microdomains to orchestrate heart rate regulation during β-AR signaling. In contrast to atrial and ventricular myocytes, SAN cells express a diverse repertoire of ACs, with ACI as the predominant Ca2+-activated isoform. Although ACI-KO (ACI-/-) mice exhibit normal cardiac systolic or diastolic function, they experience SAN dysfunction. Similarly, SAN-specific CRISPR/Cas9-mediated gene silencing of ACI results in sinus node dysfunction. Mechanistically, hyperpolarization-activated cyclic nucleotide-gated 4 (HCN4) channels form functional microdomains almost exclusively with ACI, while ryanodine receptor and L-type Ca2+ channels likely compartmentalize with ACI and other AC isoforms. In contrast, there were no significant differences in T-type Ca2+ and Na+ currents at baseline or after β-AR stimulation between WT and ACI-/- SAN cells. Due to its central characteristic feature as a Ca2+-activated isoform, ACI plays a unique role in sustaining the rise of local cAMP and heart rates during β-AR stimulation. The findings provide insights into the critical roles of the Ca2+-activated isoform of AC in sustaining SAN automaticity that is distinct from contractile cardiomyocytes.


Effect of nutritional counselling on hepatic, muscle and adipose tissue fat content and distribution in non-alcoholic fatty liver disease.
To assess the effectiveness of the current UK clinical practice in reducing hepatic fat (IHCL). Whole body MRI and (1)H MRS were obtained, before and after 6 mo nutritional counselling, from liver, soleus and tibialis muscles in 10 subjects with non-alcoholic fatty liver disease (NAFLD). A 500 Kcal-restricted diet resulted in an average weight loss of 4% (-3.4 kg,) accompanied by significant reductions in most adipose tissue (AT) depots, including subcutaneous (-9.9%), abdominal subcutaneous (-10.2%) and intra-abdominal-AT (-11.4%). Intramyocellular lipids (IMCL) were significantly reduced in the tibialis muscle (-28.2%). Decreases in both IHCL (-39.9%) and soleus IMCL (-12.2%) content were also observed, although these were not significant. Several individuals showed dramatic decreases in IHCL, while others paradoxically showed increases in IHCL content. Changes in body composition were accompanied by improvements in certain liver function tests: serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Significant correlations were found between decreases in IHCL and reductions in both intra-abdominal and abdominal subcutaneous AT. Improvements in liver function tests were associated with reductions in intra-abdominal AT, but not with changes in IHCL. This study shows that even a very modest reduction in body weight achieved through lifestyle modification can result in changes in body fat depots and improvements in LFTs.


A phase I study of EKB-569 in combination with capecitabine in patients with advanced colorectal cancer.
To determine the maximum tolerated dose (MTD), characterize the principal toxicities, and assess the pharmacokinetics of EKB-569, an oral selective irreversible inhibitor of the epidermal growth factor receptor tyrosine kinase, in combination with capecitabine in patients with advanced colorectal cancer. Patients were treated with EKB-569 daily for 21 days and capecitabine twice daily for 14 days of a 21-day cycle. The dose levels of EKB-569 (mg/day) and capecitabine (mg/m(2) twice daily) assessed were 25/750, 50/750, 50/1,000 and 75/1,000. An expanded cohort was enrolled at the MTD to better study toxicity and efficacy. Samples of plasma were collected to characterize the pharmacokinetics of the agents. Treatment efficacy was assessed every other cycle. A total of 37 patients, the majority of whom had prior chemotherapy, received a total of 163 cycles of treatment. Twenty patients were treated at the MTD, 50 mg EKB-569, daily and 1,000 mg/m(2) capecitabine twice daily. Dose-limiting toxicities were diarrhea and rash. No patients had complete or partial responses but 48% had stable disease. The conversion of capecitabine to 5-fluorouracil was higher for the combination of EKB-569 and capecitabine (321+/-151 ng*h/mL) than for capecitabine alone (176+/-62 ng*hours/mL; P=0.0037). In advanced colorectal cancer, 50 mg EKB-569 daily can be safely combined with 1,000 mg/m(2) capecitabine twice a day. A statistically significant increase in plasma levels of 5-fluorouracil for the combination of EKB-569 and capecitabine may be due to the single-dose versus multiple-dose exposure difference, variability in exposure or a potential drug interaction.


Disability adjusted life years and minimal disease: application of a preference-based relevance criterion to rank enteric pathogens.
Burden of disease estimates, which combine mortality and morbidity into a single measure, are used increasingly for priority setting in disease control, prevention and surveillance. However, because there is no clear exclusion criterion for highly prevalent minimal disease in burden of disease studies its application may be restricted. The aim of this study was to apply a newly developed relevance criterion based on preferences of a population panel, and to compare burden of disease estimates of five foodborne pathogens calculated with and without application of this criterion. Preferences for twenty health states associated with foodborne disease were obtained from a population panel (n = 107) with the Visual Analogue Scale and the Time Trade-off (TTO) technique. The TTO preferences were used to derive the relevance criterion: if at least 50% of a panel of judges is willing to trade-off time in order to be restored to full health the health state is regarded as relevant, i.e. TTO median is greater than 0. Subsequently, the burden of disease of each of the five foodborne pathogens was calculated both with and without the relevance criterion. The panel ranked the health states consistently. Of the twenty health states, three did not meet the preference-based relevance criterion. Application of the relevance criterion reduced the burden of disease estimate of all five foodborne pathogens. The reduction was especially significant for norovirus and rotavirus, decreasing with 94% and 78% respectively. Individual preferences elicited with the TTO from a population panel can be used to empirically derive a relevance criterion for burden of disease estimates. Application of this preference-based relevance criterion results in considerable changes in ranking of foodborne pathogens.


Effect of histamine-2-receptor antagonists versus sucralfate on stress ulcer prophylaxis in mechanically ventilated patients: a meta-analysis of 10 randomized controlled trials.
We conducted a meta-analysis in order to investigate the effect of histamine-2-receptor antagonists (H2RA) versus sucralfate on stress ulcer prophylaxis in mechanically ventilated patients in the intensive care unit (ICU). A systematic literature search of Medline, EMBASE, Cochrane Central Register of Controlled Trials (1966 to January 2010) was conducted using specific search terms. A review of Web of Science and a manual review of references were also performed. Eligible studies were randomized control trials (RCTs) that compared H2RA and sucralfate for the prevention of stress ulcer in mechanically ventilated patients. Main outcome measures were rates of overt bleeding, clinically important gastrointestinal (GI) bleeding, ventilator-associated pneumonia, gastric colonization and ICU mortality. Ten RCTs with 2,092 participants on mechanical ventilation were identified. Meta-analysis showed there was a trend toward decreased overt bleeding when H2RA was compared with sucralfate (OR = 0.87, 95% CI: 0.49 to 1.53). A total of 12 clinically important GI bleeding events occurred among 667 patients (1.8%) in the H2RA group compared with 26 events among 673 patients (3.9%) in the sucralfate groups. Prophylaxis with sucralfate decreased the incidence of gastric colonization (OR = 2.03, 95% CI: 1.29 to 3.19) and ventilator-associated pneumonia (OR = 1.32, 95% CI: 1.07 to 1.64). Subgroup analysis showed H2RA was not superior to sucralfate in reducing early-onset pneumonia (OR = 0.62, 95%CI: 0.36 to 1.07) but had a higher late-onset pneumonia rate (OR = 4.36, 95%CI: 2.09 to 9.09) relative to sucralfate. No statistically significant reduction was observed in mortality of ICU between groups (OR = 1.08, 95% CI: 0.86 to 1.34). In patients with mechanical ventilation, H2RA resulted in no differential effectiveness in treating overt bleeding, but had higher rates of gastric colonization and ventilator-associated pneumonia. Additional RCTs of stress ulcer prophylaxis with H2RA and sucralfate are needed to establish the net benefit and risks of adverse effect in mechanically ventilated patients.


Preclinical development of an in vivo BCG challenge model for testing candidate TB vaccine efficacy.
There is an urgent need for an immunological correlate of protection against tuberculosis (TB) with which to evaluate candidate TB vaccines in clinical trials. Development of a human challenge model of Mycobacterium tuberculosis (M.tb) could facilitate the detection of such correlate(s). Here we propose a novel in vivo Bacille Calmette-Guérin (BCG) challenge model using BCG immunization as a surrogate for M.tb infection. Culture and quantitative PCR methods have been developed to quantify BCG in the skin, using the mouse ear as a surrogate for human skin. Candidate TB vaccines have been evaluated for their ability to protect against a BCG skin challenge, using this model, and the results indicate that protection against a BCG skin challenge is predictive of BCG vaccine efficacy against aerosol M.tb challenge. Translation of these findings to a human BCG challenge model could enable more rapid assessment and down selection of candidate TB vaccines and ultimately the identification of an immune correlate of protection.


Dopamine pathway is highly diverged in primate species that differ markedly in social behavior.
In the endeavor to associate genetic variation with complex traits, closely related taxa are particularly fruitful for understanding the neurophysiological and genetic underpinnings of species-specific attributes. Similarity to humans has motivated research into nonhuman primate models, yet few studies of wild primates have investigated immediate causal factors of evolutionarily diverged social behaviors. Neurotransmitter differences have been invoked to explain the distinct behavioral suites of two baboon species in Awash, Ethiopia, which differ markedly in social behavior despite evolutionary propinquity. With this natural experiment, we test the hypothesis that genomic regions associated with monoamine neurotransmitters would be highly differentiated, and we identify a dopamine pathway as an outlier, highlighting the system as a potential cause of species-specific social behaviors. Dopamine levels and resultant variation in impulsivity were likely under differential selection in the species due to social system structure differences, with either brash or circumspect social behavior advantageous to secure mating opportunities depending on the social backdrop. Such comparative studies into the causes of the behavioral agendas that create and interact with social systems are of particular interest, and differences in temperament related to boldness and associated with dopamine variation likely played important roles in the evolution of all social, behaviorally complex animals, including baboons and humans.


Abnormal plasma fibrinolysis in patients with rheumatoid arthritis and impaired endothelial fibrinolytic response in those complicated by vasculitis.
(a) To assess plasma fibrinolytic parameters in patients with rheumatoid arthritis (RA) and to determine whether there are differences between patients with RA alone and those with RA complicated by vasculitis. (b) To determine if patients with RA respond differently to venous occlusion compared with normal subjects and to assess whether such a response differs in patients with RA alone and those with rheumatoid vasculitis. (c) To determine the extent of vascular damage in patients with rheumatoid vasculitis and if this correlates with the levels of plasma fibrinolytic parameters. Sixty three patients with RA (38 had RA only and 25 had evidence of rheumatoid vasculitis) were recruited. Plasma levels of tissue plasminogen activator antigen (t-PA Ag), plasminogen activator inhibitor (PAI) activity, and factor VIII von Willebrand factor (vWF) were measured before and 10 minutes after venous occlusion. Patients with RA, with or without rheumatoid vasculitis, had higher baseline PAI levels than control subjects. The difference was statistically significant for patients with RA alone but was not statistically significant for patients with rheumatoid vasculitis. After venous occlusion, t-PA Ag levels increased significantly in normal subjects and patients with RA alone, but not in patients with rheumatoid vasculitis. Plasma levels of vWF were significantly higher in patients with rheumatoid vasculitis than in normal subjects and those with RA alone. In patients with RA alone, baseline vWF correlated positively with t-PA Ag levels, whereas a negative correlation was found between these two parameters in patients with rheumatoid vasculitis. A negative correlation between vWF and t-PA Ag levels after venous occlusion was also found in patients with rheumatoid vasculitis. Patients with rheumatoid vasculitis showed evidence of vascular damage with increased levels of vWF and impaired t-PA Ag release after venous occlusion, a useful measurement of endothelial reserve to remove fibrin. This may be of pathophysiological importance in the development of vasculitis in these patients.


Finite element modeling of light propagation in turbid media under illumination of a continuous-wave beam.
Spatially resolved spectroscopy provides a means for measuring the optical properties of biological tissues, based on analytical solutions to diffusion approximation for semi-infinite media under the normal illumination of an infinitely small light beam. The method is, however, prone to error in measurement because the actual boundary condition and light beam often deviate from that used in deriving the analytical solutions. It is therefore important to quantify the effect of different boundary conditions and light beams on spatially resolved diffuse reflectance in order to improve the measurement accuracy of the technique. This research was aimed at using finite element method (FEM) to model light propagation in turbid media, subjected to normal illumination by a continuous-wave beam of infinitely small or finite size. Three types of boundary conditions [i.e., partial current (PCBC), extrapolated (EBC), and zero (ZBC)] were evaluated and compared against Monte Carlo (MC) simulations, since MC could provide accurate fluence rate and diffuse reflectance. The effect of beam size was also investigated. Overall results showed that FEM provided results as accurate as those of the analytical method when an appropriate boundary condition was applied. ZBC did not give satisfactory results in most cases. FEM-PCBC yielded a better fluence rate at the boundary than did FEM-EBC, while they were almost identical in predicting diffuse reflectance. Results further showed that FEM coupled with EBC effectively simulated spatially resolved diffuse reflectance under the illumination of a finite size beam. A large beam introduced more error, especially within the region of illumination. Research also confirmed an earlier finding that a light beam of less than 1 mm diameter should be used for estimation of optical parameters. FEM is effective for modeling light propagation in biological tissues and can be used for improving the optical property measurement by the spatially resolved technique.


Higher-level phylogeny of Foraminifera inferred from the RNA polymerase II (RPB1) gene.
Macroevolutionary relations among main lineages of Foraminifera have traditionally been inferred from the small subunit ribosomal genes (SSU rDNA). However, important discrepancies in the rates of SSU rDNA evolution between major lineages led to difficulties in accurate interpretation of SSU-based phylogenetic reconstructions. Recently, actin and beta-tubulin sequences have been used as alternative markers of foraminiferal phylogeny and their analyses globally confirm results obtained with SSU rDNA. In order to test new protein markers, we sequenced a fragment of the largest subunit of the RNA polymerase II (RPB1), a nuclear encoded single copy gene, for 8 foraminiferal species representing major orders of Foraminifera. Analyses of our data robustly confirm previous SSU rDNA and actin phylogenies and show (i) the paraphyly and ancestral position of monothalamid Foraminifera; (ii) the independent origin of miliolids; (iii) the monophyly of rotaliids, including buliminids and globigerinids; and (iv) the polyphyly of planktonic families Globigerinidae and Candeinidae. Additionally, the RPB1 phylogeny suggests Allogromiidae as the most ancestral foraminiferal lineage. In the light of our study, RPB1 appears as a valuable phylogenetic marker, particularly useful for groups of protists showing extreme variations of evolutionary rates in ribosomal genes.


Effects of motivational conflicts on visually elicited saccades in monkeys.
The prospect of reward evoked by external stimuli is a central element of goal-oriented behavior. To elucidate behavioral effects of reward expectation on saccade latency, we employed a visually guided saccade task with asymmetrical reward schedule. The monkey had to make an immediate saccade to a peripheral visual target in every trial, but was rewarded for a correct saccade to only one of four possible target positions. Reward availability was predictable on the basis of the spatial position of the target throughout a daily session. Compared with the condition where all positions were rewarded with a smaller amount, the mean saccade latency in the asymmetrical reward schedule was significantly shorter when the saccade was made toward the position associated with reward than when it was directed to no-reward positions. Furthermore, a divergence-point analysis on cumulative latency distributions showed that the expectation of reward facilitated saccades at all latency ranges. In contrast, the expected lack of reward delayed the initiation of saccades with latencies longer than about 200 ms, irrespective of whether the saccade was made to a position orthogonal or opposite to the reward position. For saccades with latencies of more than approximately 240 ms, an additional delay was observed when the saccade was made to a position opposite, as compared to orthogonal, to the reward position. These results suggest that the facilitation by predictive reward is mediated by a preparatory process that is location-specific, whereas the inhibition by the absence of reward takes about 200 ms after the target onset to become effective and is initially location nonspecific but turns location-specific over time.


Curcumin prevents and reverses murine cardiac hypertrophy.
Chromatin remodeling, particularly histone acetylation, plays a critical role in the progression of pathological cardiac hypertrophy and heart failure. We hypothesized that curcumin, a natural polyphenolic compound abundant in the spice turmeric and a known suppressor of histone acetylation, would suppress cardiac hypertrophy through the disruption of p300 histone acetyltransferase-dependent (p300-HAT-dependent) transcriptional activation. We tested this hypothesis using primary cultured rat cardiac myocytes and fibroblasts as well as two well-established mouse models of cardiac hypertrophy. Curcumin blocked phenylephrin-induced (PE-induced) cardiac hypertrophy in vitro in a dose-dependent manner. Furthermore, curcumin both prevented and reversed mouse cardiac hypertrophy induced by aortic banding (AB) and PE infusion, as assessed by heart weight/BW and lung weight/BW ratios, echocardiographic parameters, and gene expression of hypertrophic markers. Further investigation demonstrated that curcumin abrogated histone acetylation, GATA4 acetylation, and DNA-binding activity through blocking p300-HAT activity. Curcumin also blocked AB-induced inflammation and fibrosis through disrupting p300-HAT-dependent signaling pathways. Our results indicate that curcumin has the potential to protect against cardiac hypertrophy, inflammation, and fibrosis through suppression of p300-HAT activity and downstream GATA4, NF-kappaB, and TGF-beta-Smad signaling pathways.


Diabetes mellitus among HIV-infected individuals in follow-up care at University of Gondar Hospital, Northwest Ethiopia.
To assess the prevalence of diabetes mellitus (DM) and associated factors among HIV-infected adults in northwest Ethiopia. Hospital-based cross-sectional study. HIV clinic of the University of Gondar Hospital, Ethiopia. All HIV-infected adults who visited the HIV clinic from December 2013 to the end of February 2014 were the source population. A structured and pretested questionnaire incorporating the WHO STEPwise approach was used. A multivariate logistic regression analysis was applied to assess factors associated with DM. The overall prevalence of type 2 DM was 8% (95% CI 5.5% to 10.5%). The prevalence of DM was higher (13.2%; 95% CI 8.0% to 18.3%) among subjects receiving pre-antiretroviral treatment (pre-ART) than among those taking ART (5.1%; 95% CI 2.6% to 7.6%). Thirteen (35.1%) of the DM cases were newly identified during the study. Obesity (adjusted OR (AOR) 6.55; 1.20 to 35.8), hypertension (AOR 3.45; 1.50 to 7.90), being in the pre-ART group (AOR 4.47; 1.80 to 11.08), hypertriglyceridaemia (AOR 2.24; 1.02 to 49.5) and tertiary-level education (AOR 11.8; 2.28 to 61.4) were associated with DM. Overall DM prevalence was high, particularly among subjects in the pre-ART group. More educated, hypertensive and obese HIV-infected adults were more likely to have DM as a comorbidity. Health policy and the clinical management of HIV-infected individuals should take into account the rising DM.