Species differences in the localization of 'peripheral' type cholecystokinin receptors in rodent brain.
A comparison of cholecystokinin (CCK) receptor binding to sections of rat, mouse and guinea pig brain has been performed using 125I-Bolton Hunter CCK and the selective peripheral CCK receptor antagonist L-365,031. In both rat and mouse, 125I-Bolton Hunter CCK binding in the region of the interpeduncular nucleus (IPN) was inhibited by L-365,031 indicating that these receptors resemble CCK receptors found in peripheral tissues. In the mouse especially, dense regions of peripheral CCK receptors were detected either side of the IPN. By contrast, in the guinea pig IPN no evidence of L-365,031-sensitive binding could be found. The present reports shows that in different species, regional variations in brain CCK receptor binding occur not only in the case of classical 'brain' receptors, but also for the more discretely localised 'peripheral' type CCK receptors.


A new strategy for prophylactic surgery in BRCA women: Combined mastectomy and laparoscopic salpingo-oophorectomy with immediate reconstruction by double DIEP flap.
Prophylactic surgery remains the most effective modality for reducing both breast and ovarian cancer rate in woman at high risk, such as BRCA1 or BRCA2. Autologous breast reconstruction with bilateral deep inferior epigastric perforator (DIEP) flap allows predictable and durable results. However, existing two-step approach separating salpingo-oophorectomy and reconstruction could even make DIEP flap impossible, or make insufflation more difficult during laparoscopy. Other authors described one-step procedure but with open laparotomy. The goal of this study was to verify the feasibility of a simultaneous procedure, including laparoscopic salpingo-oophorectomy. We included BRCA mutation careers scheduled for simultaneous laparoscopic salpingo-oophorectomy, and bilateral breast reconstruction with DIEP flaps. The first step of the procedure was laparoscopic salpingo-oophorectomy and ports had to be strategically placed to avoid interference with the following procedure. The second step was bilateral breast reconstruction with DIEP flaps. We reviewed medical charts. Surgical procedure was analyzed for duration, revisions and surgical complications. During 1-year period, eight patients agreed to a simultaneous procedure. All of them were BRCA positive, mean age was 38.3years (range, 39-50), and mean BMI was 28.3kg/m(2) (range, 21-33). The mean duration of the entire procedure was 524minutes (range, 405-630) and the mean hospital stay 9.2 days (range, 8-14). There was 100% flap survival. No abdominal wall dehiscence occurred. One-step procedure for prophylactic surgery of ovarian and breast hereditary malignancies is feasible. First salpingo-oophorectomy with open laparoscopy then bilateral immediate or delayed breast reconstruction with DIEP flaps can be performed.


Incidence of preterm premature rupture of membranes and its association with inter-pregnancy interval: a prospective cohort study.
Preterm premature rupture of membranes is one of the causes of premature birth and perinatal deaths, particularly in developing countries due to poor access and availability of medical resources to manage and sustain the pregnancy to term. Although, several risk factors for preterm premature rupture of membranes were identified, its association with inter-pregnancy interval was understudied. Therefore, we aimed to assess the incidence of preterm premature rupture of membranes and its association with inter-pregnancy interval in urban South Ethiopia. A community-based prospective cohort study was conducted among 2578 pregnant women, and followed until delivery. A generalized linear model for binary outcome was applied for the analysis, using a 95% confidence level and P-value. The incidence of preterm premature rupture of membranes was 2%, 95% CI: 2%, 3%. However, the incidence was varied across the months of inter-pregnancy intervals 4% (< 18 months), 2% (18-23 months) and 1% (24-60 months). The risk of preterm premature rupture of membranes was nearly three times (ARR = 2.59, 95%CI: 1.27, 5.29) higher for women with inter-pregnancy intervals < 18 months than 24-60 months. Inter-pregnancy interval under 18 months increases the risk of preterm premature rupture of membranes, highlighting the need to improve pregnancy spacing in the community.


Impaired rectal sensation in idiopathic faecal incontinence.
In 15 patients suffering from idiopathic faecal incontinence and in 15 matched controls, manometric studies of anorectal pressure and studies of the rectoanal reflex and rectal sensitivity were carried out. Patients with idiopathic faecal incontinence had normal resting pressure but reduced squeeze and stress pressures; the anal sphincter relaxed before a sensation of rectal distension occurred. The conclusion is that both reduced voluntary muscle contraction and impairment of rectal sensation are conducive to soiling in idiopathic faecal incontinence.


Influences of elbow, shoulder, trunk motion and temporospatial parameters on arm swing asymmetry of Parkinson's disease during walking.
Arm swing asymmetry is commonly observed in early Parkinson's disease (PD) and has been found to be useful for early diagnosis. However, there are uncertainties about the nature of its relationships with gait parameters, especially shoulder and elbow motions. Therefore, this study explored how these relationships are different between PD and controls. Forty one early PD and 23 controls were included. Participants walked at self-selected speed for 3D motion analysis. Arm swing at the wrist (AS), temporospatial parameters and kinematics in elbow, shoulder and trunk were obtained. Amplitudes and asymmetries of these variables were compared between PD and control groups. PD group showed increased AS asymmetry, compared to controls. Multiple hierarchical regression analysis on AS asymmetry was conducted in order to investigate how PD influences on the relationship between AS asymmetry and other variables. In pooled data (PD and control group), asymmetries in elbow and shoulder range of motion (RoM) were significant predictors for AS asymmetry but walking speed and asymmetries in temporospatial parameters were not significant. Group effect (PD effect) was significantly mediated by only elbow RoM asymmetry. Interaction between group and elbow RoM asymmetry was statistically significant, indicating that group was an effect modifier for elbow RoM asymmetry effect on AS asymmetry. Conclusively, arm swing asymmetry measured at the wrist represents the involvement of PD effect on the unilateral and distal upper limb in early stage. These findings are helpful for future researches related to clinical applications and mechanisms of arm swing asymmetry in PD.


Mesoscale cortex-wide neural dynamics predict self-initiated actions in mice several seconds prior to movement.
Volition - the sense of control or agency over one's voluntary actions - is widely recognized as the basis of both human subjective experience and natural behavior in nonhuman animals. Several human studies have found peaks in neural activity preceding voluntary actions, for example the readiness potential (RP), and some have shown upcoming actions could be decoded even before awareness. Others propose that random processes underlie and explain pre-movement neural activity. Here, we seek to address these issues by evaluating whether pre-movement neural activity in mice contains structure beyond that present in random neural activity. Implementing a self-initiated water-rewarded lever-pull paradigm in mice while recording widefield [Ca++] neural activity we find that cortical activity changes in variance seconds prior to movement and that upcoming lever pulls could be predicted between 3 and 5 s (or more in some cases) prior to movement. We found inhibition of motor cortex starting at approximately 5 s prior to lever pulls and activation of motor cortex starting at approximately 2 s prior to a random unrewarded left limb movement. We show that mice, like humans, are biased toward commencing self-initiated actions during specific phases of neural activity but that the pre-movement neural code changes over time in some mice and is widely distributed as behavior prediction improved when using all vs. single cortical areas. These findings support the presence of structured multi-second neural dynamics preceding self-initiated action beyond that expected from random processes. Our results also suggest that neural mechanisms underlying self-initiated action could be preserved between mice and humans.


Active trans-plasma membrane water cycling in yeast is revealed by NMR.
Plasma membrane water transport is a crucial cellular phenomenon. Net water movement in response to an osmotic gradient changes cell volume. Steady-state exchange of water molecules, with no net flux or volume change, occurs by passive diffusion through the phospholipid bilayer and passage through membrane proteins. The hypothesis is tested that plasma membrane water exchange also correlates with ATP-driven membrane transport activity in yeast (Saccharomyces cerevisiae). Longitudinal (1)H(2)O NMR relaxation time constant (T(1)) values were measured in yeast suspensions containing extracellular relaxation reagent. Two-site-exchange analysis quantified the reversible exchange kinetics as the mean intracellular water lifetime (τ(i)), where τ(i)(-1) is the pseudo-first-order rate constant for water efflux. To modulate cellular ATP, yeast suspensions were bubbled with 95%O(2)/5%CO(2) (O(2)) or 95%N(2)/5%CO(2) (N(2)). ATP was high during O(2), and τ(i)(-1) was 3.1 s(-1) at 25°C. After changing to N(2), ATP decreased and τ(i)(-1) was 1.8 s(-1). The principal active yeast ion transport protein is the plasma membrane H(+)-ATPase. Studies using the H(+)-ATPase inhibitor ebselen or a yeast genetic strain with reduced H(+)-ATPase found reduced τ(i)(-1), notwithstanding high ATP. Steady-state water exchange correlates with H(+)-ATPase activity. At volume steady state, water is cycling across the plasma membrane in response to metabolic transport activity.


Ascorbate biosynthesis and its involvement in stress tolerance and plant development in rice (Oryza sativa L.).
Ascorbic acid (AsA) biosynthesis and its implications for stress tolerance and plant development were investigated in a set of rice knock-out (KO) mutants for AsA biosynthetic genes and their wild-types. KO of two isoforms of GDP-D-mannose epimerase (OsGME) reduced the foliar AsA level by 20-30%, and KO of GDP-L-galactose phosphorylase (OsGGP) by 80%, while KO of myo-inositol oxygenase (OsMIOX) did not affect foliar AsA levels. AsA concentration was negatively correlated with lipid peroxidation in foliar tissue under ozone stress and zinc deficiency, but did not affect the sensitivity to iron toxicity. Lack of AsA reduced the photosynthetic efficiency as represented by the maximum carboxylation rate of Rubisco (Vmax), the maximum electron transport rate (Jmax) and the chlorophyll fluorescence parameter ΦPSII. Mutants showed lower biomass production than their wild-types, especially when OsGGP was lacking (around 80% reductions). All plants except for KO mutants of OsGGP showed distinct peaks in foliar AsA concentrations during the growth, which were consistent with up-regulation of OsGGP, suggesting that OsGGP plays a pivotal role in regulating foliar AsA levels during different growth stages. In conclusion, our data demonstrate multiple roles of AsA in stress tolerance and development of rice.


[Aeromonas spp. infections: retrospective study in Nîmes University Hospital, 1997-2004].
Retrospective study of patients presenting Aeromonas spp. infections admitted to Nîmes hospital from January 1997 to December 2004. Aeromonas spp. infections were collected from the database of the bacteriology department. The Aeromonas species, suceptibility phenotype, epidemiological data, site and type (nosocomial or not) of infection, and evolution were collected from medical files. Thirty infections were notified corresponding to 22 male and eight female of 50.6 years old (mean) and a mean stay duration of 31 days. Skin and soft tissues (50%), digestive tract (26.7%), bones and articulations (10%), blood stream (6.7%), urinary tract (3.3%) and lymph nodes (3.3%) were the sites of infection. Ten infections were nosocomial, four were associated with medical leeches. All strains were resistants to amoxicillin and amoxicillin- clavulanic acid, whereas resistance rate to other beta-lactams was under 10% and all were suceptible to fluoroquinolones and aminoglycosides. All patients received antibiotic treatment: 31% a single molecule, 69% an association and 62% needed a second line treatment. Strain was suceptible to the antibiotic in 78.5% of cases. Only one death occured not directly linked to Aeromonas infection. Most of Aeromonas spp. infection sites were cutaneous and digestive. Nosocomial infections associated with medical leeches are not so uncommon and strict conditions of storage and administration are necessary. According to the susceptibility phenotype of our strains which is similar to literature data, a third generation cephalosporin or a fluoroquinolone should be used evenly associated with an aminoglycoside.


Occupational Exposure to Solar Radiation and the Eye: A Call to Implement Health Surveillance of Outdoor Workers.
Globally, hundreds of millions of outdoor workers are exposed to solar radiation (SR) for the majority of their working life. Such occupational exposure is known to induce various adverse health effects to the eyes, mainly related to its ultraviolet (UV) component. The present work is a call-to-action aimed at raising the awareness on the need of health surveillance for the prevention of the chronic ocular diseases in outdoor workers. Photo-chemical chronic damage can induce pterygium at the outer layer of the eye and cataract in the lens. Considering carcinogenic effects, rare squamous-cells tumors of the cornea and/or of the conjunctiva and the ocular melanomas are associated with UV radiation exposure. Solar UV-related eye diseases should be considered as "occupational diseases" when there is sufficient exposure in workers, but they are often not recognized, and/or frequently not reported to the national compensation authorities. Therefore, to prevent the burden of these work-related eye pathologies, an adequate risk assessment with identification of appropriate preventive measures, and a provision of periodic health surveillance to the exposed workers, in particular considering those at higher risk of exposure or with individual susceptibility, should be urgently implemented.


An open, prospective, randomized study comparing the immunogenicity and safety of two inactivated hepatitis A pediatric vaccines in toddlers, children and adolescents in China.
Vaccines against hepatitis A provide long-lasting protection in both adults and children. The aim of this study was to demonstrate that the inactivated hepatitis A vaccine AVAXIM 80U Pediatric is safe and not inferior in terms of seroprotection rate to HAVRIX 720 vaccine 1 month after booster vaccination. An open, randomized, single-center trial was conducted in China in healthy antihepatitis A virus seronegative individuals from 12 months to 15 years of age. Participants were randomized in a 2:1 ratio to receive either AVAXIM 80U Pediatric or HAVRIX 720, followed by a booster vaccination, using the same vaccine 6 months afterward. A total of 720 individuals were included in the study, 480 in the AVAXIM 80U Pediatric group and 240 in the HAVRIX 720 group, and 686 individuals completed the full vaccination schedule. AVAXIM 80U Pediatric was statistically noninferior to HAVRIX 720 in terms of seroprotection rate for all individuals and in each of 3 age groups: toddlers (12-23 months), children (2-11 years) and adolescents (12-15 years). Antihepatitis A virus geometric mean titers were significantly higher with AVAXIM 80U Pediatric than with HAVRIX 720. Both inactivated hepatitis A vaccines were well-tolerated and had a similar incidence and type of adverse events. AVAXIM 80U Pediatric is safe and immunogenic, with a seroprotection rate that is not inferior to HAVRIX 720 in a pediatric population of healthy individuals.


Superbranched polyglycerol nanostructures as drug delivery and theranostics tools for cancer treatment.
Hyperbranched polymers (HBPs), such as hyperbranched polyglycerols (HPGs) with a dendritic configuration, have been recognized for their excellent biocompatibility and multifunctionalization. HPGs have been studied for use in the delivery diagnostic, imaging and therapeutic molecules in the area of nanobiomedicine. They show superior characteristics to linear polymers and dendrimers, such as compact structure, a simple manufacturing process with easy functionalization ability, low viscosity, and high stability. Owing to these advantages, HPGs are now considered promising carriers for drug delivery, diagnostics, imaging, and theranostics applications for cancer treatment. In this review, we also discuss safety aspects of HPG-based nanoformulations in various animal models and the clinical translation status of such polymers for real-time applications.


Associations between cardiovascular comorbidities and mortality, length of hospital stay, and total charges among traumatic injury patients.
Cardiovascular conditions are highly prevalent and particularly common in subsets of the population at high risk for traumatic injury. This study evaluates the extent to which cardiovascular comorbidity may increase risks of negative outcomes in patients receiving trauma treatment. Clinical data for all patients admitted for traumatic injury (defined by ICD-9 diagnosis codes) of all levels of severity between the years of 2006 and 2014 in the Detroit USA metropolitan area were obtained from the State Inpatient Database for Michigan. The association between four types of cardiovascular comorbidity (hypertension, congestive heart failure, pulmonary circulation disorders, and valvular heart disease), and three outcomes (mortality, length of hospital stay, and total charges), was assessed using generalized linear modeling, both alone and after controlling for injury severity, injury region, and demographic factors. All four comorbidities examined were related to worse outcomes on all three dimensions. The greatest magnitude of estimated effects with each outcome was associated with pulmonary circulation disorders (mortality OR = 2.99, length of stay IRR = 1.69, hospital charges IRR = 1.76), and the smallest magnitude of estimated effects was associated with hypertension (mortality OR = 1.20, length of stay IRR = 1.20, hospital charges IRR = 1.18). After adjustment for the presence of multiple comorbidities, injury severity and region, age, gender, and race, all effect estimates remained significant and in the same direction, except valvular heart disease which was unrelated to mortality, and hypertension was related to lower risk of mortality (OR = 0.76). Cardiovascular comorbidities are related to higher risk of negative outcomes among patients hospitalized due to traumatic injury. Screening for these comorbidities on admission may help to improve patient outcomes.


Opposite Charge Movements Within the Photoactive Site Modulate Two-Step Channel Closing in GtACR1.
Guillardia theta anion channelrhodopsin 1 is a light-gated anion channel widely used as an optogenetic inhibitory tool. Our recently published crystal structure of its dark (closed) state revealed that the photoactive retinylidene chromophore is located midmembrane in a full-length intramolecular tunnel through the protein, the radius of which is less than that of a chloride ion. Here we show that acidic (glutamate) substitutions for residues within the inner half-tunnel enhance the fast channel closing and, for residues within the outer half-tunnel, enhance the slow channel closing. The magnitude of these effects was proportional to the distance of the mutated residue from the photoactive site. These data indicate that the local electrical field across the photoactive site controls fast and slow channel closing, involving outward and inward charge displacements. In the purified mutant proteins, we observed corresponding opposite changes in kinetics of the M photocycle intermediate. A correlation between fast closing and M rise and slow closing and M decay observed in the mutants suggests that the Schiff base proton is one of the displaced charges. Opposite signs of the effects indicate that deprotonation and reprotonation of the Schiff base take place on the same (outer) side of the membrane and explains opposite rectification of fast and slow channel closing. Оur comprehensive protein-wide acidic residue substitution screen shows that only mutations of the residues located in the intramolecular tunnel confer strong rectification, which confirms the prediction that the tunnel expands upon photoexcitation to form the anion pathway.


Expression of Bacillus licheniformis α-amylase in Pichia pastoris without antibiotics-resistant gene and effects of glycosylation on the enzymic thermostability.
Thermostable α-amylases are widely used in industry. The α-amylase from Bacillus licheniformis (BLA) with six potential glycosylation sites possessed excellent thermal and pH stability and high activity. Here, it was expressed in Pichia pastoris. The Pic-BLA-producing yeast without any antibiotics-resistant gene was cultivated in flasks and the amylase activity in fermentation supernatant reached 900 U/mL. The recombinant α-amylase Pic-BLA produced in P. pastoris was deeply glycosylated with 30% increase in molecular mass (MM). The deglycosylation treatment by Endoglycosidase H (Endo H) reduced the MM of Pic-BLA. Thermostability analysis showed that Pic-BLA and deglycosylated Pic-BLA were similar in heat tolerance. In order to eliminate the extra impact of Endo H, the BLA was also expressed in Escherichia coli to get non-glycosylated Eco-BLA. A comparative study between non-glycosylated Eco-BLA and glycosylated Pic-BLA showed no obvious difference in thermostability. It is speculated that the glycosylation has little effect on the thermostability of α-amylase BLA.


Obesity Prevention in Latin America.
In Latin American countries, obesity prevalence has increased significantly as a result of rapid urbanization and an improvement in socioeconomic conditions. We report the prevalence of overweight and/or obesity and prevention efforts in five countries: Mexico, Colombia, Brazil, Peru, and Chile. In children, the highest and lowest rates of obesity are found in Chile (23 % in 6-year-olds) and Peru (1.8 % in those <5 years), respectively. In adults, Mexico and Chile present similar high rates of obesity (around 35 %), whereas in Brazil and Colombia, the rates are around 20 % and 16.5 %, respectively. In general, the highest prevalence occurs in low-income women. Every country has developed initiatives to target obesity, from the government to the private sector and academia, mainly at the health sector and school settings. Food labeling is being addressed, but has not been implemented yet. Two interventions are described, a community-based in Mexico and a school-based in Chile. Because the increase in chronic diseases, especially diabetes, has paralleled that of obesity, effective prevention efforts are urgently needed.


Inclusion of cisplatinum (cis-DDP, Platidiam, Lachema) into combined chemotherapeutic protocol for the treatment of testicular cancer.
Fifty-seven patients with testicular tumors have been treated with combined chemotherapy (vinblastine, bleomycin and cisplatinum). By evaluation of therapeutic activity and toxic effects it was shown that Platidiam, Lachema, effectivity was identical with the two other cisplatinum preparations used in this study.


Co-chairman's remarks: genetics and the twentieth century.
Genetics, the study of the gene, developed all through the twentieth century. It began with a search for mutations in a variety of organisms and the mapping of these mutations on chromosomes. Since it applied to humans, genetics became of interest to some of the ideologies that flourished during this century. In particular, eugenic ideas were used as an argument by the Nazis for the atrocities they committed in concentration camps. The gene long remained a 'being of reason', until the middle of the century, when it turned out to be made of DNA. In 1953, the discovery of the structure of DNA gave a completely new turn to genetics.


Impacts of Dietary Nutritional Composition on Larval Development and Adult Body Composition in the Yellow Fever Mosquito (Aedes aegypti).
the mosquito Aedes aegypti (Ae. aegypti) is an important vector of arboviruses, including Zika, Dengue, and Chikungunya. The dietary requirements of larval Ae. aegypti are not well understood and likely impact developmental and physiological parameters knowledge of which could be important for vector control. This study examines the effects nutrition has on growth and development of larval Ae. aegypti of laboratory-reared Rockefeller strain mosquitoes. mosquito larvae were split into five feeding groups with diets providing different ratios of protein and carbohydrates. Each group received autolyzed Brewer's yeast (AY - high-protein), and/or rice flour (RF-high-carbohydrate). The groups were monitored to record larval developmental times, adult sizes and nutritional stores. the 100% AY group failed to pupate, suggesting the AY alone is either lacking in critical nutrients or is toxic at higher concentrations. The 100% RF group resulted in the smallest adults that took the longest time to reach pupation. Of the remaining groups, the 25% AY/75% RF (Med-low) diet yielded adult mosquitoes with highest average weight, wing length, and lipid stores relative to the other diets. the dietary requirements for development, body size, and nutrient stores of Ae. aegypti mosquitoes appear to be dependent on a relatively low but essential proportion of dietary protein to carbohydrates to achieve optimal developmental outcomes.


The effect of outreach workers' services on the medical care utilization of a disadvantaged population.
An original goal of the Kaiser-Permanente Neighborhood Health Center Project was to organize the project so that a medically indigent population would be able to utilize fully and appropriately the services of a complex medical care program. A special program of outreach services was designed as the principal means to achieve this goal. This study was made to determine the effects of these outreach services on (1) the use of or nonuse of ambulatory care services; (2) the volume and type of services used; (3) the patterns of use; and (4) the appointment-keeping behavior of the project population for a 12-month period. Outreach and medical care services were provided to an average of 7,000 persons in 1,500 low-income families who were enrolled as health plan members in the Kaiser-Permanente Medical Care Program. Project participants were randomly divided into two groups, one with and one without services, and utilization data were collected from their medical and administrative records. The findings suggest that outreach intervention had a positive effect on access to care. Persons who received outreach services were more likely to contact the medical care system; these persons also showed a substantial difference in the volume of services they used, when compared to those without outreach services. Outreach workers were less successful in changing utilization patterns, although slight differences were found in the direction of more appropriate use. Persons with outreach services were more likely to have made contacts with their regular physician, to have made a smaller proportion of walk-in contacts, to have had a higher proportion of regularly scheduled contacts, and to have made a higher proportion of continuing visits. Outreach workers also had little or no effect on appointment-keeping behavior.


Giant Cell Tumor of the Distal Radius: A Review.
Giant cell tumor of the distal radius presents a significant challenge in management due to high risk of recurrence and potential loss of function. Shared decision-making guides management, particularly for more advanced lesions. Intralesional curettage can optimize wrist function but at the cost of a higher recurrence risk. Wide resection decreases local recurrence but has higher complication rates regardless of reconstruction method. No functional difference exists between motion-preserving procedures and arthrodesis; therefore, patients should be clearly informed of the risks and benefits of each treatment option.


Transplantation of expanded endothelial colony-forming cells improved outcomes of traumatic brain injury in a mouse model.
Endothelial progenitor cells (EPCs) are critical for repairing injured tissue. Endothelial colony-forming cells (ECFCs) are a homogeneous subtype of EPCs. We investigated whether intravenously infused human ECFCs homed to injured brain promoted angiogenesis and ameliorate neurologic disabilities in a mouse model of traumatic brain injury. ECFCs were generated by in vitro propagation of EPCs from human umbilical cord blood. Young female nude mice received intravenously ECFCs from human newborns (1 × 10(6)) 1 h after they were exposed to lateral fluid percussion injury. Neurologic function was evaluated by a modified neurologic severity score and Morris water maze. ECFC homing and neovascularization at the site of injury were examined by fluorescence in situ hybridization and histochemistry on days 2 and 14 after injury, respectively. Donor ECFCs were detected in injured brain 24 h after infusion. The modified neurologic severity score and Morris water maze tests were used to evaluate neurologic disability, and found the rate of neurologic disability was improved in mice that received ECFCs. Microvessel density and expression of the proangiogenic growth factors stromal cell-derived factor-1 and vascular endothelial growth factor were also increased in the region of injured brain from mice that received ECFCs compared with those received vehicle control. These data suggest that ECFCs are effective in promoting neovascularization and improving neurologic functions after traumatic brain injury.


Structural basis of Notch O-glucosylation and O-xylosylation by mammalian protein-O-glucosyltransferase 1 (POGLUT1).
Protein O-glucosyltransferase 1/Rumi-mediated glucosylation of Notch epidermal growth factor-like (EGF-like) domains plays an important role in Notch signaling. Protein O-glucosyltransferase 1 shows specificity for folded EGF-like domains, it can only glycosylate serine residues in the C1XSXPC2 motif, and it possesses an uncommon dual donor substrate specificity. Using several EGF-like domains and donor substrate analogs, we have determined the structures of human Protein O-glucosyltransferase 1 substrate/product complexes that provide mechanistic insight into the basis for these properties. Notably, we show that Protein O-glucosyltransferase 1's requirement for folded EGF-like domains also leads to its serine specificity and that two distinct local conformational states are likely responsible for its ability to transfer both glucose and xylose. We also show that Protein O-glucosyltransferase 1 possesses the potential to xylosylate a much broader range of EGF-like domain substrates than was previously thought. Finally, we show that Protein O-glucosyltransferase 1 has co-evolved with EGF-like domains of the type found in Notch.POGLUT1 is a protein-O-glucosyltransferase that transfers glucose and xylose to the EGF-like domains of Notch and other signaling receptors. Here the authors report the structure of human POGLUT1 in complexes with 3 different EGF-like domains and donor substrates and shed light on the enzyme's substrate specificity and catalytic mechanism.


Ultrasonographic findings and natural history of intraductal papillary-mucinous neoplasms of the pancreas.
It is clear that the prevalence of malignancy is high in the main-duct type of intraductal papillary-mucinous neoplasm (IPMN). Branch-duct IPMNs include several histologic conditions such as carcinoma, adenoma, and hyperplasia. Intraductal papillary adenocarcinoma and papillary adenoma are characterized by papillary protrusions and thick septum-like structures in dilated ducts as delineated by ultrasonography. A solid mass showing a mixedecho pattern in the pancreatic parenchyma is a characteristic finding of invasive types of IPMN. The international guidelines for the management of branch-duct IPMNs suggest that the appearance of symptoms attributable to the cyst, the presence of intramural nodules, a cyst size greater than 30 mm, and dilation of the main pancreatic duct (>6 mm) are indications for resection. Based on the relationship between the height of a papillary protrusion and the diameter of a cystic dilated branch as well as on histological findings, branch-duct IPMNs with papillary protrusions more than 10 mm in height as shown by imaging should be resected, and it is not adequate to differentiate carcinoma from other lesions based on the diameter of cystic branches alone. A follow-up study on branch-duct IPMNs revealed that most papillary protrusions showed a slow increase in size or development of lateral spread, and that there was no development of cancer with stromal invasion during an average follow-up of 46 months. Therefore, the presence of intramural nodules alone should not be an indication for surgery. Also, patients without papillary protrusions or thick septum-like structures are not immediate candidates for surgery. Invasive adenocarcinoma can develop at a pancreatic site different from the area of interest showing cystic changes, with such invasion possibly being multicentric. Therefore, in patients with branch-duct IPMNs, attention should be paid to the entire pancreas when performing follow-up examinations.


Ni(II) interaction with a peptide model of the human TLR4 ectodomain.
Ni(II) stimulates innate immunity via the direct binding to human Toll Like Receptor 4 (hTLR4), the bacterial lypopolysaccharide receptor. The binding is specific for humans and causes nickel contact allergy. The protein sequence analysis of hTLR4 revealed that the ectodomain, the region supposed to coordinate the metal ions, contains a histidine-rich motif that is not conserved among all organisms. To elucidate the role of each histidine residue on the protein-nickel binding, we examined the formation of Ni(II) complexes with the model peptide NH2-FQHSNRKQMSERSVFRSRRNRIYRDISHTHTR-COO-, which encompasses the sequence 429-460 of hTLR4. The amino acid sequence of the peptide has been modified by the substitution of some selected lipophilic residues (Leu and Phe) with hydrophilic residues (Arg), aiming at increasing the peptide hydro solubility of the protein fragment. Potentiometric, ultraviolet-visible (UV-vis), nuclear magnetic resonance (NMR) and circular dichroism (CD) measurements demonstrate that the non-conserved histidines in the ectodomain cooperate in metal coordination and consequently enable the activation of the molecular mechanism of nickel hypersensitivity reaction.


Macroflux microprojection array patch technology: a new and efficient approach for intracutaneous immunization.
We evaluated the Macroflux microprojection array patch technology as a novel system for intracutaneous delivery of protein antigens. Macroflux microprojection array systems (330-microm micro-projection length, 190 microprojections/cm2, 1- and 2-cm2 area) were coated with a model protein antigen, ovalbumin (OVA), to produce a dry-film coating. After system application, microprojection penetration depth, OVA delivery, and comparative immune responses were evaluated in a hairless guinea pig model. Macroflux microprojections penetrated into hairless guinea pig skin at an average depth of 100 microm with no projections deeper than 300 microm. Doses of I to 80 microg of OVA were delivered via 1- or 2-cm2 systems by varying the coating solution concentration and wearing time. Delivery rates were as high as 20 microg in 5 s. In a prime and boost dose immune response study, OVA-coated Macroflux was most comparable to equivalent doses injected intradermally. Higher antibody titers were observed when OVA was administered with the microprojection array or intradermally at low doses (1 and 5 microg). Macroflux administration at 1- and 5-microg doses gave immune responses up to 50-fold greater than that observed after the same subcutaneous or intramuscular dose. Dry coating an adjuvant, glucosaminyl muramyl dipeptide, with OVA on the Macroflux resulted in augmented antibody responses. Macroflux skin patch technology provides rapid and reproducible intracutaneous administration of dry-coated antigen. The depth of skin penetration targets skin immune cells; the quantity of antigen delivered can be controlled by formulation, patch wearing time, and system size. This novel needle-free patch technology may ultimately have broad applications for a wide variety of therapeutic vaccines to improve efficacy and convenience of use.


Multiple Targeting of HSP Isoforms to Challenge Isoform Specificity and Compensatory Expression.
Heat shock proteins (HSPs) are molecular chaperones that assist in protein folding, trafficking, and metabolism. Intracellular chaperone functions of HSPs had been well-investigated, but extracellular and exosomal HSPs have been recently found. Exosomal HSPs are intercellularly transferred, while extracellular HSPs play cytokine-like roles called chaperokines. We have shown that exosomal HSPs play key roles in intercellular communication between tongue carcinoma and tumor-associated macrophages in the tumor microenvironment. Notably, HSP90 isoforms consist of HSP90alpha, HSP90beta, mitochondrial TRAP1, and GRP94 in the endoplasmic reticulum. Moreover, many pseudogenes of HSP90 can be transcribed into RNA. Besides, the function of HSP90 is defined by their cochaperones, such as CDC37 or AHA1. Therefore, isoform-specific small interfering RNA (siRNA) is necessary for precisely targeting each HSP90 isoform and cochaperone. Nevertheless, we often encountered compensatory expression of HSP90 isoforms in the knockdown studies. Here, we provide dual and triple knockdown methods to target multiple RNA for challenging isoform-specific roles and compensatory expression of intracellular, extracellular, and exosomal HSPs.


Hox13 is essential for formation of a sensory organ at the terminal end of the sperm duct in Ciona.
Species-specific traits are thought to have been acquired by natural selection. Transcription factors play central roles in the evolution of species-specific traits. Hox genes encode a set of conserved transcription factors essential for establishing the anterior-posterior body axis of animals. Changes in the expression or function of Hox genes can lead to the diversification of animal-body plans. The tunicate ascidian Ciona intestinalis Type A has an orange-colored structure at the sperm duct terminus. This orange-pigmented organ (OPO) is the characteristic that can distinguish this ascidian from other closely related species. The OPO is formed by the accumulation of orange-pigmented cells (OPCs) that are present throughout the adult body. We show that Hox13 is essential for formation of the OPO. Hox13 is expressed in the epithelium of the sperm duct and neurons surrounding the terminal openings for sperm ejection, while OPCs themselves do not express this gene. OPCs are mobile cells that can move through the body vasculature by pseudopodia, suggesting that the OPO is formed by the accumulation of OPCs guided by Hox13-positive cells. Another ascidian species, Ciona savignyi, does not have an OPO. Like Hox13 of C. intestinalis, Hox13 of C. savignyi is expressed at the terminus of its sperm duct; however, its expression domain is limited to the circular area around the openings. The genetic changes responsible for the acquisition or loss of OPO are likely to occur in the expression pattern of Hox13.


Increased skeletal muscle ceramide level in men at risk of developing type 2 diabetes.
Intramyocellular lipids, including ceramide, a second messenger in the sphingomyelin signalling pathway, might contribute to the development of insulin resistance. The aim of our study was to assess parameters of the skeletal muscle sphingomyelin signalling pathway in men at risk of developing type 2 diabetes. We studied 12 lean (BMI < 25 kg/m(2)) men without a family history of diabetes (control group), 12 lean male offspring of type 2 diabetic patients, and 21 men with overweight or obesity comprising 12 with NGT (obese-NGT) and nine with IGT (obese-IGT). A euglycaemic-hyperinsulinaemic clamp and a biopsy of vastus lateralis muscle were performed. Ceramide, sphingomyelin, sphinganine and sphingosine levels and sphingomyelinase and ceramidase activities were measured in muscle. Muscle diacylglycerol and triacylglycerol levels were estimated in a subgroup of 27 men (comprising men from all the above groups). Compared with the control group, the lean offspring of diabetic patients and the men with overweight or obesity showed lower insulin sensitivity (all p < 0.005) and a greater muscle ceramide level (all p < 0.01). The obese-IGT group had lower insulin sensitivity (p = 0.0018) and higher muscle ceramide (p = 0.0022) than the obese-NGT group. There was lower muscle sphingosine level and alkaline ceramidase activity in offspring of diabetic patients (p = 0.038 and p = 0.031, respectively) and higher sphinganine level in the obese-NGT (p = 0.049) and obese-IGT (p = 0.002) groups than in the control group. Muscle sphingomyelin was lower (p = 0.0028) and neutral sphingomyelinase activity was higher (p = 0.00079) in the obese-IGT than in the obese-NGT group. Muscle ceramide was related to insulin sensitivity independently of other muscle lipid fractions. Ceramide accumulates in muscle of men at risk of developing type 2 diabetes.


Delivery of double singleton pregnancies in a woman with a double uterus, double cervix, and complete septate vagina.
Uterine anomalies involving a double uterus, double cervix, also known as didelphys uterus, and complete septate vagina are rarely seen and have an associated fertility problem. However, artificial reproductive technology with embryo transfers can help solve this fertility challenge. Conception in the uterus in just one side is commonly seen for embryos, which are always transferred through the usually used (dilated) vagina. We here present a patient with the above uterine anomaly who conceived with the aid of in vitro fertilization and embryo transfer to both uterine cavities under general anesthesia, which resulted in successful double singleton pregnancies with one fetus in each uterus. With intensive prenatal care, the pregnancy course for each fetus was rather uneventful. Although both fetuses were in cephalic presentation, cesarean section was performed at the 39(th) week of gestation with good outcomes in order to preclude anticipated difficulties if the baby had been delivered through the rarely dilated vagina. However, order of birth between the two fetuses was a crucial decision during the operation.


[Familial tubulopathy associating hypercalciuria and saline diabetes. Favorable results of indomethacin therapy].
The association of hypercalciuria, salt losing renal disease, and a defect in urine concentrating ability with high renin and aldosterone levels is described in two brothers. One child had marked nephrocalcinosis by the age of 3 and the other severe growth retardation. In one child all the abnormalities were abolished with indomethacin which was responsible for a marked decrease in urinary. The data and the family study suggest that this condition is a proximal tubular disorder with atuosomal recessive inheritance.


Body mass index and mortality in patients with severe coronary artery diseases: A cohort study from China.
It is still controversial whether obesity and overweight increase the risk of mortality for patients with coronary artery disease. The current study aimed to investigate the relationship between body mass index (BMI) and mortality in patients with triple-vessel disease (TVD). From April 2004 to February 2011, 8943 patients with angiographically confirmed TVD were consecutively enrolled. Patients were divided into five groups according to BMI: underweight (<18.5 kg/m2), normal weight (18.5-23.9 kg/m2), overweight: (24-27.9 kg/m2), mild obesity (28-31.9 kg/m2), and severe obesity (≥32 kg/m2). The primary end point was all-cause death. Subgroup analysis was performed for treatment strategies: revascularization and medical treatment alone. During a median follow-up of 7.5 years, lower risks of mortality were observed in patients with overweight (adjusted HR 0.85, 95% CI 0.75-0.97) and mild obesity (adjusted HR 0.83, 95% CI 0.69-1.00) compared to those with normal weight. Polynomial Cox regression suggested a U-shape association between BMI and adjusted mortality risk. In the revascularization subgroup, there was a significantly higher mortality risk in patients with severe obesity (adjusted HR 1.57, 95% CI 1.03-2.40) than in those with normal weight. While in the medical treatment subgroup, mortality risk decreased as BMI increased, with the lowest risk being observed in patients with severe obesity. There is a U-shape relationship between BMI and all-cause death in patients with TVD, with increased risks among both underweight and severely obese patients. This relationship may be influenced by treatment strategies.


Global transcriptome analysis of rat hypothalamic arcuate nucleus demonstrates reversal of hypothalamic gliosis following surgically and diet induced weight loss.
The central mechanisms underlying the marked beneficial metabolic effects of bariatric surgery are unclear. Here, we characterized global gene expression in the hypothalamic arcuate nucleus (Arc) in diet-induced obese (DIO) rats following Roux-en-Y gastric bypass (RYGB). 60 days post-RYGB, the Arc was isolated by laser-capture microdissection and global gene expression was assessed by RNA sequencing. RYGB lowered body weight and adiposity as compared to sham-operated DIO rats. Discrete transcriptome changes were observed in the Arc following RYGB, including differential expression of genes associated with inflammation and neuropeptide signaling. RYGB reduced gene expression of glial cell markers, including Gfap, Aif1 and Timp1, confirmed by a lower number of GFAP immunopositive astrocyte profiles in the Arc. Sham-operated weight-matched rats demonstrated a similar glial gene expression signature, suggesting that RYGB and dietary restriction have common effects on hypothalamic gliosis. Considering that RYGB surgery also led to increased orexigenic and decreased anorexigenic gene expression, this may signify increased hunger-associated signaling at the level of the Arc. Hence, induction of counterregulatory molecular mechanisms downstream from the Arc may play an important role in RYGB-induced weight loss.


Readiness for hospital discharge post-initial invasive percutaneous transhepatic biliary drainage: A mixed-methods study.
This study explores perceptions regarding hospital discharge readiness among patients with post-first invasive percutaneous transhepatic biliary drainage (PTBD), family caregivers, and healthcare providers who are involved during the discharge period. A convergent mixed-method design was applied. A purposive sample of 30 patients completed a scale measuring readiness for hospital discharge, and 30 participants, including patients, family caregivers, and healthcare providers, participated in in-depth interviews. Descriptive analyses were combined with quantitative data, thematic analyses with qualitative data, and joint displays with mixed analyses. Findings indicate that readiness for hospital discharge was high, the expected support subscale was at the highest possible level, and the personal status subscale was at the lowest level. Three main themes emerged from an analysis of the interview transcripts: improved health conditions, self-care knowledge, and homecare preparedness. Self care knowledge had three sub-themes: taking care of biliary drainage, consuming a suitable diet, and observation of abnormal symptoms. Being ready for hospital discharge contributes to a safer transition from hospital to home. Healthcare providers need to reconsider the criteria for discharge and clarify patients' individual needs. Patients, family caregivers, and healthcare providers need to be prepared for hospital discharge.


Varying Degree of Overlap Following the Critical Steps of Lumbar Fusion and Short-term Outcomes.
Retrospective cohort. The present study analyzes the impact of end-overlap on short-term outcomes after single-level, posterior lumbar fusions. Few studies have evaluated how "end-overlap" (i.e., surgical overlap after the critical elements of spinal procedures, such as during wound closure) influences surgical outcomes. Retrospective analysis was performed on 3563 consecutive adult patients undergoing single-level, posterior-only lumbar fusion over a 6-year period at a multi-hospital university health system. Exclusion criteria included revision surgery, missing key health information, significantly elevated body mass index (>70), non-elective operations, non-general anesthesia, and unclean wounds. Outcomes included 30-day emergency department visit, readmission, reoperation, morbidity, and mortality. Univariate analysis was carried out on the sample population, then limited to patients with end-overlap. Subsequently, patients with the least end-overlap were exact-matched to patients with the most. Matching was performed based on key demographic variables-including sex and comorbid status-and attending surgeon, and then outcomes were compared between exact-matched cohorts. Among the entire sample population, no significant associations were found between the degree of end-overlap and short-term adverse events. Limited to cases with any end-overlap, increasing overlap was associated with increased 30-day emergency department visits ( P =0.049) but no other adverse outcomes. After controlling for confounding variables in the demographic-matched and demographic/surgeon-matched analyses, no differences in outcomes were observed between exact-matched cohorts. The degree of overlap after the critical steps of single-level lumbar fusion did not predict adverse short-term outcomes. This suggests that end-overlap is a safe practice within this surgical population.


The higher order auditory cortex is involved in the assignment of affective value to sensory stimuli.
The sensory cortex participates in emotional memory but its role is poorly understood. Here we show that inactivation of the higher order auditory cortex Te2 in rats during early memory consolidation impairs remote first- and second-order fear memories but not the association between two neutral cues. Furthermore, Te2 inactivation prevents changes in the valence of such information. Following the presentation of two auditory cues previously paired with either pleasant or painful stimuli, a large percentage of cells responds to both experiences but also a small fraction of neurons responds exclusively to one of them. The latter type of neurons signals the valence rather than the salience or the motor responses associated with the stimuli, and reflects selective associative processes. Pharmacogenetic silencing of memory-activated neurons causes amnesia. Thus, Te2 represents a crucial node for the assignment of the affective value to sensory stimuli and for the storage of such information.


Prognosis in node-negative breast cancer.
In a selected group of 207 breast cancer patients with tumor-free axillary nodes, clinical and pathological features were evaluated as to their relationship to long-term disease-free survival. No clinical feature was found to be prognostically useful. Of pathologic features studied, four appear to have significance. These are the volume of the primary mass, the histologic or nuclear grade, the presence of invasive lobular carcinoma in the primary mass, and possibly the presence of neoplastic cells in intramammary lymphatic vessels. When two or more of these four features are present, prognosis is less favorable than when there is only one, but the influences are not arithmetically additive.


Operative safety and oncologic outcomes in rectal cancer based on the level of inferior mesenteric artery ligation: a stratified analysis of a large Korean cohort.
To compare high and low inferior mesenteric artery (IMA) ligation in a large number of patients, and investigate the short-term and long-term outcomes. This retrospective study compared outcomes between high IMA ligation and low IMA ligation with dissection of lymph nodes (LNs) around the IMA origin. A total of 1,213 patients underwent elective low anterior resection with double-stapling anastomosis for stage I-III rectal cancer located ≥6 cm from the anal verge (835 patients underwent IMA ligation at the IMA origin; 378 patients underwent IMA ligation directly distal to the root of the left colic artery along with dissection of LNs around the IMA origin). There was no difference in anastomotic leakage rate between groups. The 2 groups did not significantly differ in intraoperative blood loss, perioperative complications, total number of harvested LNs, and metastatic IMA LNs. However, more metastatic LNs were harvested in the high-tie than in the low-tie group (1.3 ± 2.9 vs. 0.8 ± 1.9, P = 0.002), and the incidence of positive pathologic nodal status was higher in the high-tie group (37.9% vs. 28.6%, P = 0.001). The 5-year local recurrence-free and metastasis-free survival rates were similar between groups, as were the 5-year overall and cancer-specific survival rates. Low IMA ligation with dissection of LNs around the IMA origin showed no differences in anastomotic leakage rate compared with high IMA ligation, without affecting oncologic outcomes. High IMA ligation did not seem to increase the number of total harvested LNs, whereas the ratio of metastatic apical LNs were similar between groups.


Intravital high-frequency ultrasonography to evaluate cardiovascular and uteroplacental blood flow in mouse pregnancy.
OBJECTIVE: The objective of this study is to define the ultrasonographic changes in the cardiovascular and uteroplacental circulation of normal pregnant mice compared to non-pregnant mice using high-frequency, high-resolution ultrasonography. METHODS: Ten to twelve-week-old CD-1 mice (six non-pregnant and six pregnant animals) were used for all experiments. Vevo® 2100 (VisualSonics) was used to evaluate the cardiovascular and uteroplacental circulation physiology. Cardiac echocardiogram and uterine artery Doppler studies were performed on all animals. Pregnant animals were evaluated on embryonic day seven (E7), thirteen (E13) and eighteen (E18). Fetal heart rate and umbilical artery Doppler flows were obtained on pregnant animals. Three-dimensional ultrasonography imaging was utilized for quantification of placental volumes. All data are presented as median {10(th)-90(th) percentiles}. RESULTS: In pregnant mice on E7 compared to non-pregnant mice, there was an increase in cardiac output (p=0.008), stroke volume (p=0.002), ejection fraction, (p=0.02) and fractional shortening (p=0.02). The maternal heart rate increased throughout gestation (p= 0.009). During pregnancy, a gestational sac was clearly visible on E7. Between E13 and E18, the fetal size and fetal heart rate increased (p=0.001) and the umbilical artery peak systolic velocity increased (p <0.001). Minimal diastolic blood flow was observed in the umbilical artery on E13, which increased slightly on day E18 (p=0.01). There was also no change in the uterine artery resistance index between non-pregnant and pregnant mice. The placental volume increased between E13 and E18 (p=0.03). CONCLUSION: Several changes noted in cardiovascular and uteroplacental systems occurring during normal murine pregnancy have striking similarities to humans and can be accurately measured using newer ultrasonographic techniques. Further studies are needed to evaluate changes in these vascular beds in mouse models of diseases such as preeclampsia and intrauterine growth restriction.


Differences in racial/ethnic disparities in patient care experiences between prostate cancer survivors and males without cancer: A SEER-CAHPS study.
Our purpose was to evaluate whether racial/ethnic disparities in patient care experiences (PCEs) differ between males with prostate cancer ("PCa group") and males without cancer ("non-cancer group"). This retrospective study used 2007-2015 National Cancer Institute's Surveillance, Epidemiology and End Results registry data linked to Consumer Assessment of Healthcare Providers and Systems surveys. PCa and non-cancer groups were propensity score matched 1:5 on demographic and clinical characteristics. Differences in racial/ethnic disparities (DRD) (non-Hispanic Black [NHB], Hispanic, non-Hispanic Asian [NHA], and Other Races compared to non-Hispanic White [NHW]) in PCEs (getting needed care, getting care quickly, doctor communication, customer service, and getting needed prescription drugs) were compared between matched PCa and non-cancer groups. Per prior literature, DRD in PCE scores were categorized as small (<3), medium (≥3 but <5) or large (≥5). There were 7312 males in the PCa group and 36,559 matched males in the non-cancer group. Within each group, all racial/ethnic minority categories reported worse scores compared to NHW individuals (p < 0.05) for ≥3 PCE composite measures. Compared to PCa group, a larger NHA-NHW difference was observed in non-cancer group for getting needed care (-4.65 in PCa vs. -7.77 in non-cancer; DRD = 3.11, p = 0.029) and doctor communication (-2.46 in PCa vs. -4.85 in non-cancer; DRD = 2.38, p = 0.023). In both PCa and non-cancer groups, racial/ethnic minorities reported worse experiences compared to NHW individuals for several PCE measures. However, the difference in getting needed care and doctor communication between NHA and NHW individuals were more pronounced in non-cancer group than PCa group.


Cholesterol arachidonate as a prostaglandin precursor in adrenocortical cells.
Rat adrenocortical cells were incubated with labeled arachidonate, and the radioactivity in unesterified fatty acids was reduced by washing with 2% albumin solutions. These cells were then incubated for two hours in the absence and presence of 7.1 x 10(-10)M ACTH. During subsequent incubation of prelabeled cells with ACTH, both the mass and radioactivity of arachidonate in adrenocortical cholesteryl esters was depleted to the same extent (30--32%). The released arachidonate was in part incorporated into phospholipids, and there was also a significant increase in unesterified arachidonic acid. During this period, there was also increased incorporation of arachidonate into labeled prostaglandins. Of this increase, 92% by isotope analysis, and 88% by radioimmunoassay techniques was attributable to prostaglandins of the E pathway. These data demonstrate that prostaglandin E synthesis is specifically increased during ACTH stimulation of rat adrenocortical cells and suggest that a major source of the arachidonate substrate for this synthesis is derived from hormone-dependent hydrolysis of cortical cholesteryl esters.


Vascular architecture of melanocytic skin tumors. A quantitative immunohistochemical study using automated image analysis.
The present study examines the distribution of blood vessels in melanocytic skin tumors. Fresh frozen sections of 11 cases each of benign nevocellular nevus, primary malignant melanoma and metastatic malignant melanoma were stained with the endothelium-specific monoclonal antibody BMA 120 and evaluated by an automated image analysis system. Additionally, the proliferative activity was assessed in parallel sections using Ki 67 monoclonal antibody. There were only slight differences between the diagnostic groups as to the vascular distribution in the tumor center, but there were remarkable differences in the connective tissue at the base of the lesions: The area occupied by small vessels (minimum diameter less than 20 microns) was 0.3 +/- 0.05% in benign nevi, 0.6 +/- 0.05% in primary malignant melanoma, and 1.2 +/- 0.10% in metastatic malignant melanoma (U-test: p less than or equal to 0.05). The proliferative activity within each lesion showed a strong positive correlation with the number of small vessels at the base of the tumor (linear regression analysis: r = 0.86; p less than or equal to 0.0001). The findings demonstrate that neovascularization in malignant melanocytic tumors takes place predominantly in the surrounding host tissue and is closely related to the proliferative activity.


Vaccination practices in pediatric transplantation: A survey among member centers of the European reference network TransplantChild.
There is considerable variation in vaccination practices between pediatric transplant centers. This study aims to evaluate active immunization attitudes and practices among ERN-TransplantChild centers and identify potential areas of improvement that could be addressed by shared evidence-based protocols. A cross-sectional questionnaire of attitudes and practices toward immunization of pediatric SOT and HSCT candidates and recipients was sent to a representative member of multidisciplinary teams from 27 European centers belonging to the ERN-TransplantChild. A total of 28/62 SOT programs and 6/12 HSCT programs across 21 European centers participated. A quarter of centers did not have an on-site protocol for the immunizations. At the time of transplantation, pediatric candidates were fully immunized (80%-100%) in 57% and 33% of the SOT and HSCT programs. Variations in the time between vaccine administration and admission to the waiting list were reported between the centers, with 2 weeks for inactivated vaccines and variable time (2-4 weeks) for live-attenuated vaccines (LAVs). Almost all sites recommended immunization in the post-transplant period, with a time window of 4-8 months for the inactivated vaccines and 16-24 months for MMR and Varicella vaccines. Only five sites administer LAVs after transplantation, with seroconversion evaluated in 80% of cases. The immunization coverage of European pediatric transplant recipients is still inconsistent and far from adequate. This survey is a starting point for developing shared evidence-based immunization protocols for safe vaccination among pediatric transplant centers and generating new research studies.


Influence of lactate to acetate ratio on biological production of medium chain carboxylates via open culture fermentation.
Waste valorisation via biological production of widely used in the industry medium chain carboxylates (MCCs) via open culture fermentation (OCF) could be a promising alternative to the commonly used anaerobic digestion. Lactate-rich waste streams are considered as valuable substrates for carboxylate chain elongation (CE), however, there are certain limitations related to the production efficiency. Acetate produced and accumulated in the acetogenesis plays an important role in CE, i.e. acetate is elongated to butyrate and then to caproate which is most popular MCC. Henceforth, it was investigated whether the ratio of lactate to acetate (L:A) affected carboxylates yields and product distribution in the lactate-based CE in OCF. The tested L:A ratios influenced carboxylates selectivity in batch trials. In the ones with lactate as the sole carbon source, propionate production was predominant but when a higher relative acetate concentration was used, the production of butyrate and CE to caproate was favored. The co-utilization of lactate and acetate in a continuous process increased the production of butyrate and caproate compared to the phase with lactate as the sole carbon source, however, controlling the relative concentration of lactate and acetate during co-utilization was not an effective strategy for increasing caproate production. 16S rRNA gene amplicon reads mapping to Caproiciproducens were the most abundant in samples collected throughout the continuous processes regardless of the L:A ratios.


Peroxiredoxin 6 mediates protective function of astrocytes in Aβ proteostasis.
Disruption of β-amyloid (Aβ) homeostasis is the initial culprit in Alzheimer's disease (AD) pathogenesis. Astrocytes respond to emerging Aβ plaques by altering their phenotype and function, yet molecular mechanisms governing astrocytic response and their precise role in countering Aβ deposition remain ill-defined. Peroxiredoxin (PRDX) 6 is an enzymatic protein with independent glutathione peroxidase (Gpx) and phospholipase A2 (PLA2) activities involved in repair of oxidatively damaged cell membrane lipids and cellular signaling. In the CNS, PRDX6 is uniquely expressed by astrocytes and its exact function remains unexplored. APPswe/PS1dE9 AD transgenic mice were once crossed to mice overexpressing wild-type Prdx6 allele or to Prdx6 knock out mice. Aβ pathology and associated neuritic degeneration were assessed in mice aged 10 months. Laser scanning confocal microscopy was used to characterize Aβ plaque morphology and activation of plaque-associated astrocytes and microglia. Effect of Prdx6 gene dose on plaque seeding was assessed in mice aged six months. We show that hemizygous knock in of the overexpressing Prdx6 transgene in APPswe/PS1dE9 AD transgenic mice promotes selective enticement of astrocytes to Aβ plaques and penetration of plaques by astrocytic processes along with increased number and phagocytic activation of periplaque microglia. This effects suppression of nascent plaque seeding and remodeling of mature plaques consequently curtailing brain Aβ load and Aβ-associated neuritic degeneration. Conversely, Prdx6 haplodeficiency attenuates astro- and microglia activation around Aβ plaques promoting Aβ deposition and neuritic degeneration. We identify here PRDX6 as an important factor regulating response of astrocytes toward Aβ plaques. Demonstration that phagocytic activation of periplaque microglia vary directly with astrocytic PRDX6 expression level implies previously unappreciated astrocyte-guided microglia effect in Aβ proteostasis. Our showing that upregulation of PRDX6 attenuates Aβ pathology may be of therapeutic relevance for AD.


[Patient-centred and culturally competent: good health care for immigrant patients requires specific competencies].
Good care for all patients, including immigrants, means patient-centred personal care by a physician with the specific knowledge and skills necessary for this individual patient and his specific context. This requires cultural competences, i.e. the knowledge and skills to handle ethnic, social and cultural health differences. These competences should be addressed more in medical education and postgraduate training and in policies regarding quality of care.


Positive effect of total knee arthroplasty on progression of arteriosclerosis evaluated by cardio-ankle vascular index.
Physical function is expected to improve with an increase in physical activity owing to improvement in knee joint pain after total knee arthroplasty (TKA). This study was performed to evaluate the impact of TKA on arteriosclerosis by measuring the cardio-ankle vascular index (CAVI) before and after surgery. In total, 206 consecutive patients undergoing unilateral TKA were investigated. The CAVI, an index of the overall stiffness of the artery from the origin of the aorta to the ankle, was used to evaluate the degree of arteriosclerosis. The CAVI of the TKA side and non-TKA side was compared before and 1 year after TKA. There were no differences in the CAVI before and after TKA on the TKA side and non-TKA side, although these values should have worsened at 1 year compared with preoperative values. The CAVI, which did not differ between the two sides preoperatively, differed significantly between the two sides postoperatively (p = 0.013). A generalized linear model showed no interaction between each time point and the measured sides in terms of the CAVI. The relationship between the preoperative CAVI and the difference between the preoperative and postoperative CAVI were examined, showing that R = - 0.428 (p < 0.001) for the TKA side and R = - 0.416 (p < 0.001) for the non-TKA side (significant negative correlation). The lack of significant age-related deterioration over time on both sides suggests that TKA may slow the progression of arteriosclerosis, especially on the operated side. The effect of TKA was found to be greater with a higher CAVI (i.e., more advanced arteriosclerosis).


Passive anion transport through the chloroplast inner envelope membrane measured by osmotic swelling of intact chloroplasts.
It has been shown that chloride channels are located in the envelope membranes of chloroplasts [5,11]. In this report, we use the light-scattering technique to measure quantitatively the rate of anion transport through the inner envelope membrane of isolated intact chloroplasts. Our results permit to assign the anion transport to the inner envelope of chloroplasts. The anionic selectivity determined from the kinetics of light scattering indicates that the chloride pathway is also highly permeable for NO-2 and NO-3. The sulfate and phosphate anions are impermeant. The chloride flux is not inhibited by DIDS or NEM and is temperature-dependent. The activation energy of the transport process suggests that the Cl- flux occurs through a channel.


The history, science, and art of wine and the case for health benefits: perspectives of an oenophilic cardiovascular pathologist.
This article stems from the intersection of the author's long-standing interests in science, medicine, pathology and oenology. A discussion is provided of selected aspects of the science of viticulture and wine production as well as qualities of the finished product crafted by the art of the wine maker. The case for health benefits of moderate consumption of wine and other alcoholic beverages also is discussed. Based on the "French paradox," an analysis is presented of the evidence for the special effects of red wine consumption, particularly as part of the Mediterranean diet and lifestyle. A concluding perspective is given in support of wine as a promoter of civility and social engagement.


The oxygen-conserving dive reflex re-examined as the principal contributory factor in sudden infant death.
This hypothesis and overview of sudden infant death syndrome (SIDS) literature is an extension of a previously published hypothesis by the author, which linked the prone sleeping position and other triggers to the oxygen-conserving dive reflex as a primary cause of SIDS. Recent prospective epidemiologic and biochemical studies, together with other data, seem to lend support to the theory that this reflex, either induced or occurring spontaneously, may lead to an irreversible pathophysiologic mechanism resulting in respiratory failure and cardiac arrest. It will also be argued that SIDS is multifactorial and tri-phasic and that several other well-known hypotheses may form a previously unrecognized part of the theory.


Targeted delivery of miR-218 via decorated hyperbranched polyamidoamine for liver cancer regression.
Hepatocellular carcinoma (HCC) is one of most common causes of cancer death worldwide. MicroRNA (miRNA) replacement gene therapy is a novel approach for HCC management. MiR-218 is a promising tumor suppressor miRNA that is down-regulated in HCC. Here, our aim was the targeted delivery of miR-218 expressing DNA plasmid (pmiR-218) to suppress HCC in vitro and in vivo. Hyperbranched polyamidoamine was synthesized via simple and economically one-pot reaction followed by decoration with lactobionic acid (LA-PAMAM) to selectively deliver and restore miR-218 expression in HCC. In vitro cytotoxicity investigations revealed the high biocompatibility of LA-PAMAM. Furthermore, decoration of hyperbranched polymer with LA moieties enabled LA-PAMAM to deliver pmiR-218 more efficiently to HepG2 cells compared to both PMAMA and naked pmiR-218. Such efficient delivery of miR-218 resulted in suppression of HepG2 proliferation and down-regulation of its oncogenic HOXA1 target. In vivo, LA-PAMAM/pmiR-218 treatment of HCC induced by DEN and CCl4 in mice leads to an obvious decrease in the number and size of HCC nodules. In addition, LA-PAMAM/pmiR-218 significantly improved the liver histological features, as well as down-regulated the HOXA1 in liver tissue. In conclusion, this study showed the potential of LA-PAMAM carrier for the targeted delivery of tumor suppressor miR-218 as a therapeutic candidate for HCC.


Emerging pathways in the development of chondrosarcoma of bone and implications for targeted treatment.
Chondrosarcoma is a malignant cartilage-forming tumour of bone, of which distinct clinicopathological subtypes are known. Conventional chondrosarcoma is notorious for its locally aggressive behaviour as well as for its resistance to chemotherapy and radiotherapy; so far surgery is the only effective therapeutic option. During the past 10 years, substantial new insights have been gained about molecular cell biology, molecular cytogenetics, and immunopathology, leading to better understanding of chondrosarcoma development at the molecular level, which will ultimately lead to better clinical understanding and possibly to the development of targeted treatment.


Non-productive infection of human newborn blood mononuclear cells with herpes simplex virus: effect on T cell activation, IL-2 production and proliferation.
Proliferative responses by T cells from newborn cord blood stimulated with PHA or CD3 were reduced following infection with live (but not killed) herpes simplex virus in vitro although activation (measured by calcium flux) and IL-2 production were unaffected. The impairment of proliferation was not reversed by exogenous IL-2, phosphonoacetic acid, indomethacin or anti-alpha or anti-gamma interferon antibodies. HSV DNA was detected by hybridization in DNA extracted from unseparated MNC and from subsets sorted for CD3+ and for HLA DR+ expression. HSV DNA replication was not detected by thymidine uptake and only small amounts of virus were recovered in an infectious centre assay, suggesting that infection was non-permissive. Nevertheless, in-vitro synthesis of a limited range of HSV proteins including ICP4 was detected by metabolic labelling.


A novel approach for gene therapy: engraftment of fibroblasts containing the artificial chromosome expression system at the site of inflammation.
Rheumatoid arthritis is characterized by inflammation of the synovial tissue. High systemic doses are necessary to achieve therapeutic levels of anti-rheumatic drugs in the joints. Gene transfer might provide a more efficient delivery system for genes encoding therapeutic proteins. The artificial chromosome expression system (ACE System) is a new non-integrating, non-viral gene expression system which functions like a natural chromosome. This technology offers advantages over current expression systems because it allows stable and predictable expression of proteins encoded by single or multiple genes over long periods of time. We are developing ex vivo gene therapy using murine artificial chromosomes containing a reporter gene (LacZ and red fluorescent protein (RFP)) for local delivery of genes in rats with adjuvant arthritis (AA). The delivery of the intact ACE System into rat fibroblast-like synoviocytes (FLS) and rat skin fibroblasts (RSF) was detected within 24 to 48 h post-transfection. After growing cells under selection, clones expressing LacZ and RFP were identified. Furthermore, we investigated the feasibility of local delivery of a reporter gene to the joints of rats with AA by ex vivo gene therapy. This resulted in engraftment of the injected cells in the synovial tissue microarchitecture and expression of the reporter gene. This work demonstrates the potential feasibility of treating arthritis and other inflammatory diseases using fibroblasts containing the ACE System as a non-viral vector for gene therapy.


Collateral Ligament Reconstruction of the Proximal Interphalangeal Joint.
Proximal interphalangeal joint collateral ligament injuries are common; however, chronic instability of this joint is rare. In such cases, however, there is no consensus on optimal management. Various repairs and reconstructions have been devised, although the literature on outcomes remains scant. We present a method of reconstruction of the proximal interphalangeal joint collateral ligament using a distally based slip of the flexor digitorum superficialis tendon.


Glycoproteins of axonal transport. Interaction with heparin.
Rapidly transported fucose-containing glycoproteins, in axons and nerve terminals, from retinal ganglion cells of the rabbit have been characterized via affinity chromatography on a new fucose-specific lectin from Aleuria aurantia. It was found that a relatively small number of highly labelled transported polypeptides did not bind to heparin-Sepharose. These polypeptides also showed a low affinity for the dye Coomassie brilliant blue following sodium dodecyl sulphate electrophoresis.


Temperature responses of carbon monoxide and hydrogen uptake by vegetated and unvegetated volcanic cinders.
Ecosystem succession on a large deposit of volcanic cinders emplaced on Kilauea Volcano in 1959 has resulted in a mosaic of closed-canopy forested patches and contiguous unvegetated patches. Unvegetated and unshaded surface cinders (Bare) experience substantial diurnal temperature oscillations ranging from moderate (16 °C) to extreme (55 °C) conditions. The surface material of adjacent vegetated patches (Canopy) experiences much smaller fluctuations (14-25 °C) due to shading. To determine whether surface material from these sites showed adaptations by carbon monoxide (CO) and hydrogen (H(2)) consumption to changes in ambient temperature regimes accompanying succession, we measured responses of CO and H(2) uptake to short-term variations in temperature and long-term incubations at elevated temperature. Based on its broader temperature optimum and lower activation energy, Canopy H(2) uptake was less sensitive than Bare H(2) uptake to temperature changes. In contrast, Bare and Canopy CO uptake responded similarly to temperature during short-term incubations, indicating no differences in temperature sensitivity. However, during extended incubations at 55 °C, CO uptake increased for Canopy but not Bare material, which indicated that the former was capable of thermal adaptation. H(2) uptake for material from both sites was completely inhibited at 55 °C throughout extended incubations. These results indicated that plant development during succession did not elicit differences in short-term temperature responses for Bare and Canopy CO uptake, in spite of previously reported differences in CO oxidizer community composition, and differences in average daily and extreme temperatures. Differences associated with vegetation due to succession did, however, lead to a notable capacity for thermophilic CO uptake by Canopy but not Bare material.


Synthesis of 4-Alkylated 1,4-Dihydropyridines: Fe(II)-Mediated Oxidative Cascade Cyclization Reaction of Cyclic Ethers with Enaminones.
Syntheses of highly functionalized 4-alkylated 1,4-dihydropyridines (1,4-DHPs) from cyclic ethers and enaminones via iron(II)-mediated oxidative free radical cascade C(sp3)-H bond functionalization/C(sp3)-O bond cleavage/cyclization reaction have been first developed. This novel synthetic strategy offers an alternative method for the construction of 1,4-DHPs by using esters as the C4 sources, as well as expands the application of ethers in heterocycle synthesis.


Statins and anti-inflammatory therapies for subarachnoid hemorrhage.
Aneurysmal subarachnoid hemorrhage (SAH) induces a potent inflammatory cascade that contributes to endothelial dysfunction, imbalance of vasoactive substances (excess endothelin, depletion of nitric oxide), and arterial vasospasm. This process results in delayed cerebral ischemia, a major cause of neurologic disability in those surviving the initial hemorrhage. The only therapy shown to be effective in improving neurologic outcomes after SAH is a calcium-channel antagonist, nimodipine (although it achieved this result without reducing vasospasm). A number of novel therapies have been explored to inhibit the development of vasospasm and reduce the burden of ischemia and cerebral infarction. Statins are promising candidates, as they block multiple aspects of the inflammatory pathway that contributes to ischemic brain injury. Early clinical trials have produced conflicting results, however, and the adoption of statins in clinical practice should await the results of larger, more definitive studies. Though endothelin-receptor antagonists showed promise in significantly reducing vasospasm in preliminary trials, their failure to improve clinical outcomes in phase 3 studies has been disappointing, highlighting the complex link between vasospasm and ischemia. Future directions in the quest to improve outcomes of patients with SAH may need to approach ischemia as a multifactorial process with inflammatory, vasoactive, and ionic/metabolic components.


[Non-immune hydrops fetalis. Review of 11 cases].
Eleven cases of non-immune hydrops fetalis (NIHF) are presented. Incidence was 1.8 out of 10,000 births. NIHF became more common than immune hydrops fetalis (1.4/10,000 births). Mortality was of 81% and complications were frequent. NIHF is associated with prematurity, neonatal anoxia, polyhydramnios, hypoalbuminemia and hypoproteinemia. Considerable emphasis must be placed on antenatal diagnosis to achieve a precocious treatment and so improve the present poor prognosis.


[Loosening of osteosynthesis screw as a rare cause of haemorrhagic pleurisy].
The occurrence of haemorrhagic pleurisy in patients with myeloma is most often related to a non-specific cause. Pleural myeloma is rare but this diagnosis should be excluded. We here report a rare case of haemorrhagic pleurisy due to the loosening of an osteosynthesis screw. The study involved a 55-year-old female patient who had been followed up for multiple myeloma since 2012. Diagnosis was confirmed by the presence of haemorrhagic pleural effusion, by chest computerized tomography scan which showed the loosening of left-sided osteosynthesis screw with tip located in the prevertebral region at the level of the pleural effusion and by negative etiological assesment.


Somatic mosaicism due to a reversion variant causing hemi-atrophy: a novel variant of dystrophinopathy.
We describe a case of hemi-atrophy in a young adult male, with a positive family history of three maternal uncles with Duchenne muscular dystrophy (DMD). The patient showed progressive weakness localized to the left side, an abnormal electromyography, and creatine kinase levels >3000 IU/l. Muscle biopsy showed both dystrophin-positive and -negative myofibers. An out-of-frame duplication variant in DMD, that is, c.(93+1_94-1)_(649+1_650-1)dup(p.?) resulting in duplication of exons 3-7 was inherited, but the muscle biopsy showed dystrophin mRNA with and without the duplication. Dystrophin quantification using mass spectrometry showed 25% normal dystrophin protein levels in the muscle biopsy from the stronger right side. Sex chromosome aneuploidy was ruled out. We conclude that the patient inherited the duplication variant, but early in development an inner cell mass underwent a somatic recombination event removing the duplication and restoring dystrophin expression. To our knowledge, this is the first report of a reversion leading to somatic mosaicism in DMD.


Inhibition by monoclonal anticomplement receptor type 1 on interactions between senescent human red blood cells and monocytic-macrophagic cells.
The effect of different murine monoclonal antibodies (Mab) specific for the glycoprotein complement receptor type 1 (CR1), type 2 (CR2), and type 3 (CR3) on the adhesion to and on the phagocytosis of human senescent red blood cells (S-RBC) by monocytes or by monocyte-derived macrophages (M phi) was investigated. Murine Mab anti-CR3 (anti-Leu 15 and OKM1) were found to inhibit, in the same order of magnitude, on one hand, the Fc receptors (FcR)-dependent rosetting and phagocytosis, and, on the other hand, the S-RBC rosetting and phagocytosis by adherent monocytes. Thus, the specific involvement of the CR3 epitopes recognized by Mab anti-Leu 15 or by OKM1 in the interactions between S-RBC and monocyte/macrophage could not be demonstrated. Murine Mab anti-CR1 was found to be a significant inhibitor of binding to and of phagocytosis of S-RBC (but not of young [Y] RBC) by monocytes or M phi, whereas Mab OKM5 carrying the same isotype as Mab anti-CR1, but a different specificity, was devoid of any significant inhibitory effect. Furthermore, Y-RBC or S-RBC opsonized with Mab anti-CR1 did not form FcR-dependent rosettes and were not internalized by monocytes; in addition, preincubation of phagocytes with Mab anti-CR1 did not inhibit FcR-dependent rosetting and phagocytosis. These results suggest that the effect of anti-CR1 is mediated through a specific binding to CR1 and not through an FcR blockade. As the role of specifically bound IgG on phagocytosis of human S-RBC by macrophages has previously been demonstrated by several authors, the present study suggests that monocyte-macrophage complement receptor type 1 may act in synergy with Fc receptors in the recognition of S-RBC by macrophages. It is shown in addition that the tripeptide Arg-Gly-Asp, identical to the region of iC3b recognized by CR3 and by several adhesion-promoting receptors that are structurally similar to CR3, such as fibronectin or vitronectin, is a significant inhibitor of the binding to and the phagocytosis of S-RBC by monocytic-macrophagic cells.


[Preparation and characterization of monoclonal antibodies against Micrococcus luteus Rpf domain].
To express Micrococcus luteus Rpf domain in prokaryotic cells and prepare monoclonal antibodies against Rpf domain. The gene encoding Micrococcus luteus Rpf domain was amplified from genome of Micrococcus luteus by polymerase chain reaction(PCR), and inserted into cloning vector pUC-19. After sequenced, Micrococcus luteus Rpf domain gene was subcloned into the expression vector pPro-EXHT and transfected into E.coli DH5alpha. After induced by IPTG, the bacteria controlled by T7 promoter expressed the fused Micrococcus luteus Rpf domain protein with a hexahistidine tail at its N-terminal and the target protein was purified under denaturing conditions. Using this protein as antigen to immunize the BALB/c mice and prepare monoclonal antibodies against Micrococcus luteus Rpf domain. Then specifities and relative affinities of mAbs were identified by ELISA. The fusion protein was purified by metal chelate affinity chromatography under denaturing condition. Three cloned mAbs were prepared from the mice immunized by Rpf domain. All of them could recognize Rpf domain. specifically. The prepared mAbs against Rpf domain have strong specificity with high titers, which provides useful tools for further study of the function of Rpf domain in TB prevention.


Genetic counseling in sickle cell disease: Insights from the Indian tribal population.
Sickle cell disease (SCD) is an inherited disorder of hemoglobin. With an overall prevalence of 4.3%, India is the second-largest hub of SCD after Africa. Genetic counseling (GC) is the most cost-effective intervention to reduce the burden of a genetic disease including SCD. Even though GC's role in reducing SCD birth prevalence is well established, it is still not incorporated into Indian national policy and is unavailable to most Indians approaching their marriageable age and childbirth. GC perception and efficacy have also not been explored yet among young adults, especially in Indian tribal communities. Counseling in these communities requires careful consideration of their socioeconomic, cultural, and ethical values. Community engagement with local tribes and healthcare infrastructure in a multitier approach is essential for an effective GC. This review aims to provide healthcare providers and genetic counselors with the essentials of GC in the prevention and management of SCD among tribal communities based on the author's counseling experience in South India.


Cyclin-dependent kinase inhibitor p27Kip1, but not p21WAF1/Cip1, is required for inhibition of hypoxia-induced pulmonary hypertension and remodeling by heparin in mice.
Heparin has growth inhibitory effects on pulmonary artery smooth muscle cell (PASMC) in vitro and in vivo. However, the mechanism has not been fully defined. In this study, we investigated the role of cyclin-dependent kinase inhibitors, p21(WAF1/cip1) (p21) and p27Kip1 (p27), in the inhibitory effect of heparin on PASMC proliferation in vitro and on hypoxia-induced pulmonary hypertension in vivo using p21 and p27-null mice. In vitro, loss of the p27 gene negated the inhibitory effect of heparin on PASMC proliferation, but p21 was not critical for this inhibition. In vivo, heparin significantly inhibited the development of hypoxia-induced pulmonary hypertension and remodeling, as evidenced by decreased right ventricular systolic pressure, ratio of right ventricular weight to left ventricle plus septum weight, and percent wall thickness of pulmonary artery, in p21(+/+), p21(-/-), p27(+/+), and p27(+/-), but not in p27(-/-) mice. We also observed that hypoxia decreased p27 expression significantly in mouse lung, which was restored by heparin. Heparin inhibited Ki67 proliferative index in terminal bronchial vessel walls in p27(+/+) and p27(+/-), but not in p27(-/-) mice exposed to hypoxia. Therefore, we conclude that the cyclin-dependent kinase inhibitor p27, but not p21, is required for the inhibition of hypoxic pulmonary vascular remodeling by heparin.


An integrative review of nursing leadership in Saudi Arabia.
The aims of this integrative review were to describe leadership styles from the nursing literature in Saudi Arabia and to identify the current state of evidence about relationships between leadership styles and nurse, patient and organization outcomes in Saudi Arabia. Integrative review was used as a design for this study. The following search terms were used with databases: 'Saudi Arabia', 'leadership theory*', 'leadership style*', 'leadership model*', 'management style' and 'nurse*'. Methodological quality was assessed using two different quality rating tools for quantitative and qualitative studies. Databases used for this review included Nursing & Allied Health Database, Cochrane Database of Systematic Reviews, PubMed, CINAHL, Embase, PsycINFO, Scopus, Web of Science and ProQuest Dissertations & Theses. Nine manuscripts representing eight studies were included in this review. The papers reviewed included quantitative (n = 6), qualitative (n = 2) and mixed methods studies (n = 1). Results were grouped into different themes, identified as nursing leadership styles in Saudi Arabia, leadership styles and nurses' outcomes, and demographics and leadership styles.


Antifungal activity of altenusin isolated from the endophytic fungus Alternaria sp. against the pathogenic fungus Paracoccidioides brasiliensis.
Altenusin is a biphenyl derivative isolated from different species of fungi, which presents several biological activities. We report the antifungal activity of the altenusin isolated from the endophytic fungus Alternaria sp., against clinical isolates of Paracoccidioides brasiliensis, and its action on cell walls of P. brasiliensis and the nonpathogenic yeast Schizosaccharomyces pombe. In vitro antifungal activity of altenusin was evaluated using the broth microdilution method against 11 strains of P. brasiliensis and one strain of S. pombe. The effects of the altenusin on the cell wall were estimated using the sorbitol protection assay. The altenusin presented strong activity against P. brasiliensis with MIC values ranging between 1.9 and 31.2 μg/ml, and 62.5 μg/ml for S. pombe. Our results demonstrated that the MIC values for altenusin were increased for P. brasiliensis Pb18 and for S. pombe when the medium was supplemented with sorbitol. Additionally, S. pombe cells treated with altenusin were more rounded in shape than untreated cells. Altenusin showed activity against clinical strains of P. brasiliensis at the concentration tested, and this compound probably affects fungal cell walls. These findings suggest that altenusin could act through the inhibition of cell wall synthesis or assembly in P. brasiliensis and S. pombe, and could be considered as a lead compound for the design of new antifungals.


From tumor biology to clinical Pet: a review of positron emission tomography (PET) in oncology.
Cancer cells show increased metabolism of both glucose and amino acids, which can be monitored with 18F-2-deoxy-2-fluoro-D-glucose (FDG), a glucose analogue, and 11C-L-methionine (Met), respectively. FDG uptake is higher in fast-growing than in slow-growing tumors. FDG uptake is considered to be a good marker of the grade of malignancy. Several studies have indicated that the degree of FDG uptake in primary lung cancer can be used as a prognostic indicator. Differential diagnosis of lung tumors has been studied extensively with both computed tomography (CT) and positron emission tomography (PET). It has been established that FDG-PET is clinically very useful and that its diagnostic accuracy is higher than that of CT. Detection of lymph node or distant metastases in known cancer patients using a whole-body imaging technique with FDG-PET has become a good indication for PET. FDG uptake may be seen in a variety of tissues due to physiological glucose consumption. Also FDG uptake is not specific for cancer. Various types of active inflammation showed FDG uptake to a certain high level. Understanding of the physiological and benign causes of FDG uptake is important for accurate interpretation of FDG-PET. In monitoring radio/chemotherapy, changes in FDG uptake correlate with the number of viable cancer cells, whereas Met is a marker of proliferation. Reduction of FDG uptake is a sensitive marker of viable tissue, preceding necrotic extension and volumetric shrinkage. FDG-PET is useful for the detection of recurrence and for monitoring the therapeutic response of tumor tissues in various cancers, including those of the lung, colon, and head and neck. Thus, PET, particularly with FDG, is effective in monitoring cancer cell viability, and is clinically very useful for the diagnosis and detection of recurrence of lung and other cancers.


Pharmacokinetics of intravenous amoxicillin in pediatric patients.
Pharmacokinetic parameters were obtained from 14 infants and children receiving intravenous amoxicillin. Peak serum values increased proportionally to the increase in dose; the serum half-life was similar in the three dose groups studied.


Description (reporting the actual words used in written letters) of the neuropsychological and psychopathological modifications produced by dopaminergic treatment in a young patient with Parkinson's disease.
Although it is an undeniable fact that dopaminergic therapy has greatly improved the quality of life and prognosis of patients with Parkinson's disease, various and serious adverse events correlated to dopaminergic drugs are not uncommon. Among these, those of neuropsychological and psychopathological nature are of particular importance, being capable of causing an upheaval in the basic personality of the patient. To this regard, the authors report the actual words of a patient himself that are far more convincing that any considerations we may express. In our opinion, the overwhelming impact of dopaminergic treatment on the psychopathological and neuropsychological equilibrium of all parkinsonian patients should unfailingly be carefully evaluated and pondered, particularly in the early onset forms of the disease.


Organized breast screening improves reattendance compared to physician referral: a case control study.
The Ontario Breast Screening Program (OBSP) is a population-based breast screening programme, not requiring physician referral. OBSP invites women by mail to book their next screens. However, women who do not participate in the OBSP, may be referred by physicians to non-OBSP mammography facilities, which do not remind women to book their next screen. We identified women without breast cancer prior to June 30, 2011, having bilateral mammography (M) during a baseline period at age 50 - 69 at OBSP or non-OBSP facilities, and during a re-exposure period, at the same facility type. We used a case-control design to study the association of facility type and having M during an outcome period. Cases were women failing to receive the outcome M. Controls were matched by age, census tract, and socioeconomic status. Exposure was baseline facility type. Covariates were comorbidity, residential mobility, and primary care physician (PCP) characteristics. Conditional logistic regression analysis was performed. Cases were less likely to have been screened at OBSP facilities. Failure to receive the outcome M was associated with having moved after re-exposure M (OR = 1.61, 95% confidence interval (CI) 1.52, 1.71), having a male PCP (OR = 1.05, 95% CI 1.02, 1.05), or a higher Charlson score (OR = 1.06 per unit increase, 95% CI 1.03, 1.09). Having re-exposure M at an OBSP facility (OR = 0.18, 95% CI 0.18, 0.19)., having a Canadian trained PCP (OR = 0.83, 95% CI 0.8, 0.87), and having a PCP one year after the re-exposure M (OR = 0.81, 95% CI 0.68, 0.97) were protective against failure to receive the outcome M. The OBSP, not requiring physician referral, and inviting women by mail to book their next screen, is associated with a lower probability of failure to reattend for subsequent screening than screening by PCP referral to non-OBSP facilities.


A microbiological assay for sterigmatocystin, T-2 toxin and zearalenone.
A survey was done to find microorganisms useful for assaying sterigmatocystin; T-2 toxin and zearalenone.Staphylococcus aureus was found to be sensitive to T-2 toxin and zearalenone;Bacillus cereus was found to be sensitive to T-2 toxin only; andEscherichia coli was sensitive to sterigmatocystin. The response of the organisms to sterigmatocystin; T-2 toxin and zearalenone was found to be linear between 4 and 100 μg with sterigmatocystin toE. coli; between 2 and 25 μg with T-2 toxin toStaph, aureus andB. cereus; and between 4 and 100 μg with zearalenone toStaph, aureus. The lower limits of sensitivity of the test were 2 μg T-2 toxin and zearalenone, and 4 μg sterigmatocystin. The assay is rapid (15-17 hrs); simple and inexpensive; and can be used to verify the toxicity of samples and to confirm thin layer chromatographic results.