The naso-axial line: a new method of accurately predicting the inferior limit of the endoscopic endonasal approach to the craniovertebral junction.
The endoscopic endonasal approach (EEA) has developed as an emerging surgical corridor to the craniovertebral junction (CVJ). In addition to understanding its indications and surgical anatomy, the ability to predict its inferior limit is vital for optimal surgical planning. To develop a method that accurately predicts the inferior limit of the EEA on the CVJ radiologically and to compare this with other currently used methods. Predissection computerized tomographic scans of 9 cadaver heads were used to delineate a novel line, the naso-axial line (NAxL), to predict the inferior EEA limit on the upper cervical spine. A previously described method with the use of the nasopalatine line (NPL or Kassam line) was also used. On computerized tomographic scans obtained following dissection of the EEA, the predicted inferior limits were compared with the actual extent of dissection. The postdissection inferior EEA limit ranged from the dens tip to the upper half of the C2 body, which matched the limit predicted by NAxL, with no statistically significant difference between them. In contrast to the NAxL, the NPL predicted a significantly lower EEA limit (P < .001), ranging from the lower half of the C2 body to the superior end plate of C3. The novel NAxL more accurately predicts the inferior limit of the EEA than the NPL. This method, which can be easily used on preoperative sagittal scans, accounts for variations in patients' anatomy and can aid surgeons in the assessment of the EEA to address caudal CVJ pathology.


Pressure ulcers--prophylaxis and treatment.
Pressure sores are common and affect 10-20% of all institutionalized patients. Health personnel should have sufficient knowledge of prevention, evaluation and treatment. This article is based on a non-systematic search in Medline and EMBASE, and the authors' own clinical experience. The causes of pressure sores are mechanical pressure, shear and frictional forces on the skin and underlying tissue. Neuropathy, nutrition deficiency, moist skin, and infection are risk factors for development and persistence of the wound. Preventive measures and treatment must to be directed towards both causes and risk factors. Pressure sores are graded on a scale from 0 to 4, based on depth of wound penetration into underlying tissue. Superficial wounds are treated conservatively while deeper pressure sores, which affect muscle and bone, must be evaluated for surgical treatment. Alleviation of pressure and reduction of risk factors are important for prevention and treatment of pressure sores. Depending on the depth of wound penetration into underlying tissue, additional treatment may be conservative wound treatment or surgery.


A 5-kg time-resolved luminescence photometer with multiple excitation sources.
A luminescence photometer was developed based on lanthanide-sensitized luminescence to detect environmental pollutants and residues in foods including, in particular, two classes of antibiotics: tetracyclines and fluoroquinolones. Multiple excitation sources, a xenon flashlamp and ultraviolet light-emitting diodes (UV LEDs), were used to their advantages. A photomultiplier tube module, gated to reject time-domain interferences, was used as a photodetector. Using danofloxacin as a model analyte, luminescence signal was linear in more than three decades (0.5-2000 ng/mL) with R(2) > 0.998 in each decade. This photometer achieved a limit of detection of 2.0 ng/mL with 1.3% average relative standard deviation. It is field deployable at 4.6 kg and 15 W power consumption.


Effects of dietary treatment on bone mineral density in adults with celiac disease: factors predicting response.
Conflicting evidence is reported about the effect of treatment on bone mineral density (BMD) in adults with celiac disease (CD). This study analyzed the effects on BMD induced by treatment with a calcium-rich, gluten-free diet in adults with nonsilent CD. In 30 women and 11 men with newly diagnosed CD, BMD was measured at the right femur (femoral neck and right Ward's triangle) and the lumbar spine by dual-energy x-ray absorptiometry under untreated conditions (pretreatment) and after 1-yr treatment with a calcium-rich, gluten-free diet. On average, posttreatment BMD was greater than pretreatment BMD at the lumbar spine (mean +/- SE: 0.907 +/- 0.028 and 0.795 +/- 0.028 g/cm2, respectively; p < 0.001), the femoral neck (0.818 +/- 0.023 and 0.741 +/- 0.030 g/cm2, respectively; p = 0.002), and the Ward's triangle (0.703 +/- 0.025 and 0.654 +/- 0.025 g/cm2, respectively; p < 0.001). The greatest BMD change (percent of baseline) was observed at the lumbar spine (+14.1%), the smallest at the Ward's triangle (+7.5%). In the absence of appropriate controls, the BMD change expected in the patients under untreated conditions was estimated by regressing pretreatment BMD over duration of CD with control for gender and age at which CD became clinically evident. The regression coefficient of this analysis indicated that 1 yr of untreated CD was associated with a BMD decrease at the lumbar spine by 0.00570 g/cm2 (95% confidence interval -0.0103 to -0.0011 g/cm2). The 95% confidence interval of the treatment-induced change in BMD at the lumbar spine (+0.060 to +0.160 g/cm2) did not overlap the 95% confidence interval of the BMD change expected under untreated conditions. A large interindividual variability was observed in the BMD response to the treatment: in univariate and multivariate analyses, the treatment-induced change in BMD was significantly related to gender (greater in women than in men) and to pretreatment age and BMD. The data show that BMD is increased by dietary treatment of CD in most but not all adult patients; pretreatment BMD, gender, and pretreatment age predict the bone response after a 1-yr treatment.


IKP104-induced decay of tubulin: role of the A-ring binding site of colchicine.
Tubulin, the major subunit protein of microtubules, has a tendency to lose its ability to assemble or to interact with ligands in a time-dependent process known as decay. Decay involves the increase in exposure of sulfhydryl groups and hydrophobic areas. The antimitotic drug IKP104 [2-(4-fluorophenyl)-1-(2-chloro-3, 5-dimethoxyphenyl)-3-methyl-6-phenyl-4(1H)-pyridinone] accelerates the decay of tubulin [Ludueña et al. (1995) Biochemistry 34, 15751-15759]. In the presence of colchicine, however, IKP104 stabilizes tubulin against decay. We have shown that the stability and the acceleration of the decay of tubulin are mediated respectively by the high- and low-affinity binding site(s) of IKP104 [Chaudhuri et al. (1998) J. Protein Chem. 17, 303-309]. To better understand the mechanism by which colchicine protects tubulin from IKP104-induced decay, we examined the effect of colchicine and its analogues on this process. We found that IKP104 unfolds tubulin in a process involving a specific domain where colchicine interacts, although the binding sites of these two drugs are distinctly different. 2-Methoxy-5-(2',3',4'-trimethoxyphenyl) tropolone (MTPT), the bicyclic analogue of colchicine that lacks the B-ring, can also protect tubulin from IKP104-induced decay. An A-ring analogue of colchicine, 3,4,5-trimethoxybenzaldehyde (TMB), can also stop IKP104-induced unfolding of tubulin significantly. Interestingly, the C-ring analogue of colchicine, tropolone methyl ether (TME), does not prevent this process. Our results thus suggest that neither the B-ring nor the C-ring binding regions of colchicine are involved in the IKP104-induced decay and that the A-ring binding site of colchicine on tubulin plays a crucial role in IKP104-induced decay.


Systemic AAV8-mediated delivery of a functional copy of muscle glycogen phosphorylase (Pygm) ameliorates disease in a murine model of McArdle disease.
McArdle disease is a disorder of carbohydrate metabolism that causes painful skeletal muscle cramps and skeletal muscle damage leading to transient myoglobinuria and increased risk of kidney failure. McArdle disease is caused by recessive mutations in the muscle glycogen phosphorylase (PYGM) gene leading to absence of PYGM enzyme in skeletal muscle and preventing access to energy from muscle glycogen stores. There is currently no cure for McArdle disease. Using a preclinical animal model, we aimed to identify a clinically translatable and relevant therapy for McArdle disease. We evaluated the safety and efficacy of recombinant adeno-associated virus serotype 8 (rAAV8) to treat a murine model of McArdle disease via delivery of a functional copy of the disease-causing gene, Pygm. Intraperitoneal injection of rAAV8-Pygm at post-natal day 1-3 resulted in Pygm expression at 8 weeks of age, accompanied by improved skeletal muscle architecture, reduced accumulation of glycogen and restoration of voluntary running wheel activity to wild-type levels. We did not observe any adverse reaction to the treatment at 8 weeks post-injection. Thus, we have investigated a highly promising gene therapy for McArdle disease with a clear path to the ovine large animal model endemic to Western Australia and subsequently to patients.


Intrinsic local disorder and a network of charge-charge interactions are key to actinoporin membrane disruption and cytotoxicity.
Actinoporins are a family of sea anemone proteins that bind to membranes and produce functional pores which result in cell lysis. Actinoporin variants with decreased lytic activity usually show a reduced affinity for membranes. However, for some of these mutant versions there is no direct correlation between the loss of binding affinity and the decrease in their overall lytic activity, suggesting that other steps in pore formation may be hampered or facilitated by the mutations. To test this hypothesis on the mechanism of pore formation by this interesting family of proteins, structural and dynamic NMR studies have been carried out on two disabled variants of the actinoporin Sticholysin II, R29Q and Y111N. It is shown that their lytic activity is not only related to their membrane affinity but also to their conformational mechanism for membrane insertion. Alterations in their activities can be explained by structural, electrostatic and dynamic differences in a cluster of aromatic moieties and the N-terminus. In addition, the dynamic properties of some segments located at the C-terminus of the R29Q variant suggest a relevant role for this region in terms of protein-protein interactions. On the basis of all these results, we propose that R29 anchors a network of electrostatic interactions crucial for the actinoporin's approach to the membrane and that Y111 induces a necessary disorder in the loop regions that bind to membranes.


Role of alpha-adrenoceptors in control of plasma potassium in conscious rabbits.
1. In conscious fed rabbits the intravenous infusion of amidephrine (10 micrograms kg-1 min-1), an alpha 1-adrenoceptor agonist, caused a significant increase in plasma potassium levels that was blunted by prazosin (50 micrograms kg-1, s.c.). Idazoxan failed to modify this response. 2. Clonidine (2 micrograms kg-1 min-1, i.v.), an alpha 2-adrenoceptor agonist, also evoked a hyperkalaemic response which was antagonized by idazoxan (1 microgram kg-1, s.c.), yohimbine (0.45 mg kg-1, s.c.) and prazosin (50 micrograms kg-1, s.c.). Apamin (40 micrograms kg-1, i.v. bolus) also suppressed the clonidine-mediated hyperkalaemia. 3. Verapamil (5 micrograms kg-1, s.c.) prevented both alpha 1- and alpha 2-adrenoceptor-mediated increase in plasma potassium levels. 4. It is concluded that in conscious fed rabbits both alpha 1- and alpha 2-adrenoceptor stimulation induce hyperkalaemia by activation of hepatic Ca(2+)-activated K(+)-channels.


beta-Endorphin: hyperthermia in mice by intravenous injection.
Effects of intravenous beta-endorphin on body temperature and body weight loss were studied in naive and morphine-dependent mice. beta-Endorphin at doses 2.6-25.5 mg/kg injected intravenously caused hyperthermia in naive mice as well as in morphine-dependent mice. In addition, beta-endorphin and morphine reduced body-weight loss during the morphine withdrawal.


A hyperspectral fluorescence system for 3D in vivo optical imaging.
In vivo optical instruments designed for small animal imaging generally measure the integrated light intensity across a broad band of wavelengths, or make measurements at a small number of selected wavelengths, and primarily use any spectral information to characterize and remove autofluorescence. We have developed a flexible hyperspectral imaging instrument to explore the use of spectral information to determine the 3D source location for in vivo fluorescence imaging applications. We hypothesize that the spectral distribution of the emitted fluorescence signal can be used to provide additional information to 3D reconstruction algorithms being developed for optical tomography. To test this hypothesis, we have designed and built an in vivo hyperspectral imaging system, which can acquire data from 400 to 1000 nm with 3 nm spectral resolution and which is flexible enough to allow the testing of a wide range of illumination and detection geometries. It also has the capability to generate a surface contour map of the animal for input into the reconstruction process. In this paper, we present the design of the system, demonstrate the depth dependence of the spectral signal in phantoms and show the ability to reconstruct 3D source locations using the spectral data in a simple phantom. We also characterize the basic performance of the imaging system.


[Development of a patient-reported outcome scale for stable chronic obstructive pulmonary disease and its clinical applicability].
To develop a patient-reported outcome (PRO) scale for patients with stable chronic obstructive pulmonary disease (sCOPD) and to study its clinical applicability in traditional Chinese medicine (TCM). According to standardized procedures, the test version of PRO scale for sCOPD (sCOPD-PRO scale) was developed based on the theory of "unity of body and spirit" and was used to test 324 patients selected from six clinical centers. Five methods were used to analyze the items; the reliability, validity and responsiveness of the remaining items of the scale were evaluated. (1) sCOPD-PRO consisted of 23 items after item analysis, and according to the content of each item, these items could be grouped into four domains, namely, lung and kidney deficiency symptom, spleen deficiency symptom, functional activities and emotional impact. (2) Cronbach's alpha coefficient for sCOPD-PRO scale was more than 0.75 in four domains and total scores. (3) sCOPD-PRO scale and St. George's Respiratory Questionnaire (SGRQ) were moderately correlated or highly correlated in corresponding domains: the correlation coefficient between the total score of sCOPD-PRO scale and the total score of SGRQ was 0.72 (P<0.01), the correlation coefficient between functional activities domain of sCOPD-PRO scale and activity domain of SGRQ was 0.77 (P<0.01), and the correlation coefficient between emotional impact domain of sCOPD-PRO scale and impact domain of SGRQ was 0.51 (P<0.01). Factor analysis revealed that four factors could explain 65.06% of the total variance. (4) The four domains and total scores of sCOPD-PRO scale were significantly different before and after intervention in TCM group and Western medicine group (P<0.01); moreover, the effect size and standardized response mean of two groups were more than 0.4. The development process and method of sCOPD-PRO scale, which is based on the theories of "unity of body and spirit", are standard, and the reliability, validity and responsiveness of the scale are sound and can be relied upon for sCOPD clinical efficacy evaluation of TCM.


Current Status of Epidemiology, Diagnosis, Therapeutics, and Vaccines for the Re-Emerging Human Monkeypox Virus.
Monkeypox (Mpox) virus, a member of the Poxviridae family, causes a severe illness similar to smallpox, which is characterized by symptoms such as high fever, rash, and pustules. Human-to-human transmission cases have been reported but remained low since the first recorded case of human infection occurred in the Congo in 1970. Recently, Mpox has re-emerged, leading to an alarming surge in infections worldwide since 2022, originating in the United Kingdom. Consequently, the World Health Organization (WHO) officially declared the '2022-23 Mpox outbreak'. Currently, no specific therapy or vaccine is available for Mpox. Therefore, patients infected with Mpox are treated using conventional therapies developed for smallpox. However, the vaccines developed for smallpox have demonstrated only partial efficacy against Mpox, allowing viral transmission among humans. In this review, we discuss the current epidemiology of the ongoing Mpox outbreak and provide an update on the progress made in diagnosis, treatment, and development of vaccines for Mpox.


Development of multi-layered gastric floating tablets based on konjac glucomannan: a modified calcium supplement with enhanced bioavailability.
This study developed a gastric-floating delivery system of calcium based on konjac glucomannan (KGM). The developed calcium tablets, consisting of one core layer coated with two barrier layers, were fabricated with a facile method. The role of KGM within the tablets was evaluated from characteristics including swelling behavior, hydrated gel properties, floating ability and release profiles of calcium (Ca). The results indicated that upregulating the KGM fraction accelerated the formation of a more compact gel network in gastric conditions, which prolonged both the floating lag time and floating duration, which resulted in a more sustained swelling behavior and a slower release of calcium. Among all the formulations, a core tablet containing 20% KGM (K20) was selected as the optimized one as it could quickly float up in 7.21 s, exhibited an almost linear release and obtained a release amount of 87.73% within 12 h. Finally, a comparison of in vivo calcium bioavailability between a KGM-based calcium tablet and a commercial calcium tablet (Caltrate®) was carried out by monitoring the serum calcium concentration after administration in rabbits. The results suggested that, after having the KGM-based calcium tablet, the changes of serum calcium levels were gentler due to a sustained-release property. The difference integral value between profile K20 and the baseline was 1.4358, larger than that of Caltrate® (1.1808), suggesting the higher absorption efficiency of KGM-based calcium tablets.


Long-term outcome of endocrine function in patients with neurohypophyseal germinomas.
Since neurohypophyseal germinomas occur at the pituitary and hypothalamic axis in children and adolescents, the endocrinopathy is one of the common and critical QOL determinants. We carried out a retrospective study on the outcome of endocrine function in patients with neurohypophyseal germinoma, in order to improve or preserve pituitary function after treatment. Sixteen patients (7 men and 9 women), aged 6 to 26 years were admitted and followed up for 95.3 (14-197) months. DI was noted in 12 patients in pretreatment and 16 in posttreatment regardless of tumor size. We carried out the replacement of GH in all 8 patients, presenting the symptoms under 15 years of age. Gonadal or gonadotropic, thyroid and adrenal hormones were replaced in 9, 12 and 15 patients, respectively. Patients with large tumor compressing chiasm or hypothalamus needed hormonal replacement such as gonadal or gonadotropic and thyroid hormones more frequently (<0.01) than those with small one. In addition, two patients with a small tumor at the pituitary stalk and the 3rd ventricle floor showed the improvement of secretion pattern in gonadotropins and ACTH after chemotherapy, although they later needed radiation therapy to control the tumor. Based on our study and review of literature, the endocrinological studies before and after treatment demonstrated that pituitary dysfunction present before treatment persisted or worsened even after tumor remission, except for patients with small and localized ones. The poor endocrine results is considered to be largely radiation-related. Chemotherapy alone seems to be insufficient to obtain complete response (CR). To avoid radiation related pituitary injury, combination of 24 Gy or less dosage of radiation and appropriate chemotherapy is essential. The earlier diagnosis by repeatedly using neuroimaging and serum and CSF tumor markers and earlier initiation of treatment, before irreversible pituitary-hypothalamic damage occurs, contributes to improvement of the outcome of pituitary functions in patients with neurohypophyseal germinomas.


Enabling animal rabies diagnostic in low-access areas: Sensitivity and specificity of a molecular diagnostic test from cerebral tissue dried on filter paper.
Rabies is a lethal zoonotic encephalomyelitis that causes an estimated 59,000 human deaths yearly worldwide. Although developing countries of Asia and Africa bear the heaviest burden, surveillance and disease detection in these countries is often hampered by the absence of local laboratories able to diagnose rabies and/or the difficulties of sample shipment from low-access areas to national reference laboratories. Filter papers offer a convenient cost-effective alternative for the sampling, shipment, and storage of biological materials for the diagnosis of many pathogens including rabies virus, yet the properties of diagnostic tests using this support have not been evaluated thoroughly. Sensitivity and specificity of molecular diagnosis of rabies infection using a reverse transcription followed by a hemi-nested polymerase chain reaction (RT-hn-PCR) either directly on brain tissue or using brain tissue dried on filter paper were assessed on 113 suspected field animal samples in comparison to the direct fluorescent antibody test (FAT) recommended by the World Health Organization as one of the reference tests for rabies diagnosis. Impact of the duration of the storage was also evaluated. The sensitivity and the specificity of RT-hn-PCR i) on brain tissue were 96.6% (95% CI: [88.1-99.6]) and 92.7% (95% CI: [82.4-98.0]) respectively and ii) on brain tissue dried on filter paper 100% (95% CI: [93.8-100.0]) and 90.9% (95% CI: [80.0-97.0]) respectively. No loss of sensitivity of RT-hn-PCR on samples of brain tissue dried on filter paper left 7 days at ambient temperature was detected indicating that this method would enable analyzing impregnated filter papers sent to the national reference laboratory at ambient temperature within a 1-week shipment time. It could therefore be an effective alternative to facilitate storage and shipment of samples from low-access areas to enhance and expand rabies surveillance.


Highly Electroactive Ni Pyrophosphate/Pt Catalyst toward Hydrogen Evolution Reaction.
Robust electrocatalysts toward the resourceful and sustainable generation of hydrogen by splitting of water via electrocatalytic hydrogen evolution reaction (HER) are a prerequisite to realize high-efficiency energy research. Highly electroactive catalysts for hydrogen production with ultralow loading of platinum (Pt) have been under exhaustive exploration to make them cutting-edge and cost-effectively reasonable for water splitting. Herein, we report the synthesis of hierarchically structured nickel pyrophosphate (β-Ni2P2O7) by a precipitation method and nickel phosphate (Ni3(PO4)2) by two different synthetic routes, namely, simple cost-effective precipitation and solution combustion processes. Thereafter, Pt-decorated nickel pyrophosphate and nickel phosphate (β-Ni2P2O7/Pt and Ni3(PO4)2/Pt) were prepared by using potassium hexachloroplatinate and ascorbic acid. The fabricated novel nickel pyrophosphate and nickel phosphate/Pt materials were utilized as potential and affordable electrocatalysts for HER by water splitting. The detailed electrochemical studies revealed that the β-Ni2P2O7/Pt (1 μg·cm-2 Pt) electrocatalyst showed excellent electrocatalytic performances for HER in acidic solution with an overpotential of 28 mV at -10 mA·cm-2, a Tafel slope of 32 mV·dec, and an exchange current density ( j0) of -1.31 mA·cm-2, which were close to the values obtained using the Vulcan/Pt (8.0 μg·cm-2 Pt), commercial benchmarking electrocatalyst with eight times higher Pt amount. Furthermore, the β-Ni2P2O7/Pt electrocatalyst maintains an excellent stability for over -0.1 V versus RHE for 12 days, keeping j0 equal after the stability test (-1.28 mA cm-2). Very well-distributed Pt NPs inside the "cages" on the β-Ni2P2O7 structure with a crystalline pattern of 0.67 nm distance to the Ni2P2O7/Pt electrocatalyst, helping the Volmer-Tafel mechanism with the Tafel reaction as a major rate-limiting step, help to liberate very fast the Pt sites after HER. The high electrocatalytic performance and remarkable durability showed the β-Ni2P2O7/Pt material to be a promising cost-effective electrocatalyst for hydrogen production.


Radiologic and endoscopic diagnosis of duodenal angioma.
Two patients with lymphangioma and one with hemangioma of the duodenum are described. The radiologic presentation of duodenal angiomas is that of multiple submucous, soft, polypoid, non-infiltrating tumors. During propagation of peristalsis, on compression, or gas distension of the duodenum, they change in shape and dimensions. Deep peristaltic waves could cause apparent vanishing of the angiomas. Possible mechanisms of such behaviour of angiomas are discussed. Listed characteristics enabled the radiologic distinction of angiomas from solid submucous duodenal tumors. Duodenoscopy allows differentiation of duodenal lymphangiomas from hemangiomas and duodenal varices. Final diagnoses were based on histologic analysis of surgical specimens.


Atypical squamous cells, cannot exclude high grade squamous intraepithelial (ASC-H) in HIV-positive women.
South Africa has very high rates of both HIV infection and cervical pathology. The management of ASC-H is colposcopy and directed biopsy, but with so many women diagnosed with HSIL and a dearth of colposcopy centres in South Africa, women with cytologic diagnosis of ASC-H may not be prioritized for colposcopy. The aim of this study was to determine if HIV-positive women with a cytologic diagnosis of ASC-H should undergo immediate colposcopy or whether colposcopy can be delayed, within the context of an underfunded health care setting with so many competing health needs. A computer database search was performed from the archives of an NGO-administered clinic that offers comprehensive HIV care. All women with a cytologic diagnosis of ASC-H on cervical smears from September 2005 until August 2009 were identified. Histologic follow up was sought in all patients. A total of 2111 cervical smears were performed and 41 diagnosed as ASC-H (1.94%). No histologic follow up data was available in 15 cases. Follow up histologic results were as follows: three negative (11.5%), five koilocytosis and/ or CIN1 (19.2%), ten CIN2 (38.5%) and eight CIN3 (30.8%). There were no cases of invasive carcinoma on follow up. The current appropriate management of HIV-positive women in low-resource settings with a diagnosis of ASC-H on cervical smear is colposcopy, despite the costs involved. In the future and if cost-effective in developing nations, use of novel markers may help select which HIV-positive women can be managed conservatively and which ones referred for more active treatment. More research in this regard is warranted.


Inflammation and stroke: the Leiden 85-Plus Study .
Experimental evidence indicates that interleukin-10 (IL-10) deficiency is associated with the development of cardiovascular and cerebrovascular disease. We analyzed the relation between low IL-10 production levels, history of stroke, and incident fatal stroke. All 85-year-old inhabitants of Leiden, Netherlands (n=599) were visited at their place of residence (response rate, 87%). Production levels of the anti-inflammatory cytokine IL-10 were assessed in a whole blood assay whereby lipopolysaccharide was used as a stimulus. Plasma concentrations of C-reactive protein (CRP) were also used as a marker of inflammation. A history of stroke was obtained at baseline (prevalence, 10%). The number of fatal strokes was prospectively obtained for a median follow-up of 2.6 years (incidence, 1.82 per 100 person-years at risk). Subjects with a history of stroke had significantly lower median IL-10 production levels at baseline than subjects without stroke (558 versus 764 pg/mL; P<0.05). They also had significantly higher median CRP concentrations (6 versus 3 mg/L; P<0.05). The odds ratio for a history of stroke increased to 2.30 (95% CI, 1.12 to 4.72) over strata representing decreasing production levels of IL-10. The relative risk for incident fatal stroke was 2.94 (95% CI, 1.01 to 8.53) when we compared subjects with low or intermediate baseline IL-10 production levels to those with high production levels of IL-10. Our data support the hypothesis that subjects with low IL-10 production levels have an increased risk of stroke.


COVID-19 severity correlates with airway epithelium-immune cell interactions identified by single-cell analysis.
To investigate the immune response and mechanisms associated with severe coronavirus disease 2019 (COVID-19), we performed single-cell RNA sequencing on nasopharyngeal and bronchial samples from 19 clinically well-characterized patients with moderate or critical disease and from five healthy controls. We identified airway epithelial cell types and states vulnerable to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In patients with COVID-19, epithelial cells showed an average three-fold increase in expression of the SARS-CoV-2 entry receptor ACE2, which correlated with interferon signals by immune cells. Compared to moderate cases, critical cases exhibited stronger interactions between epithelial and immune cells, as indicated by ligand-receptor expression profiles, and activated immune cells, including inflammatory macrophages expressing CCL2, CCL3, CCL20, CXCL1, CXCL3, CXCL10, IL8, IL1B and TNF. The transcriptional differences in critical cases compared to moderate cases likely contribute to clinical observations of heightened inflammatory tissue damage, lung injury and respiratory failure. Our data suggest that pharmacologic inhibition of the CCR1 and/or CCR5 pathways might suppress immune hyperactivation in critical COVID-19.


[Stroke mimics: a challenge for the emergency physician].
To study the number of patients diagnosed with a stroke in the emergency service of a first level hospital and the proportion of these patients who were finally stroke mimics, as well as to describe and analyze the variables that can help in differential diagnosis in hospital emergency services. Nine month prospective study. All patients evaluated in emergency services and admitted with a diagnosis of stroke were included. Different clinical variables were analyzed that included prior history, history of the current event, general physical and neurological examination, NIHSS classification and Oxfordshire Community Stroke Project Classification. The final diagnosis was made by a panel of experts with access to clinical characteristics, image studies and other tests. The univariate and multivariate analysis determined the characteristics that help in distinguishing strokes from stroke mimics. One hundred and forty-four cases of stroke were registered; the final sample was made up of 140 patients. The final diagnosis was stroke in 103 out of 140 (73.6%) and stroke mimic in the rest. Eleven variables predicted the diagnosis in patients with a suspected stroke: age over 70 years (p=0.012), NIHSS classification > 5, reaching a clinical classification (p=0.019) capable of determining the exact start (p=0.000), abnormal vascular findings (p=0.014), gaze deviation (p=0.042), sight loss (p=0.052) and extensor plantar response (p=0.025) favored diagnosis of stroke, while epileptic seizures (p=0.029), neurological symptoms not congruent with the vascular territory (p=0.022) and abnormal findings in other systems (p=0.14) favored diagnosis of stroke mimic. Stroke mimics constitute one-third of the patients admitted from emergency services as strokes, with a highly varied etiology. Achieving a suitable clinical history and a precise physical examination is of great importance and can help less experienced doctors.


Determination of primary and secondary aliphatic amines with high performance liquid chromatography based on the derivatization using 1,3,5,7-tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)-difluoroboradiaza-s-indacene.
In this article, the simultaneous determination of primary and secondary aliphatic amines including dimethylamine (DMA), diethylamine and eleven primary aliphatic amines by high performance liquid chromatography (HPLC) with fluorescence detection has been achieved using a BODIPY-based fluorescent derivatization reagent, 1,3,5,7-tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)-difluoroboradiaza-s-indacene (TMBB-Su). The derivatization reaction of TMBB-Su with aliphatic amines was optimized with orthogonal design experiment and the derivatization reaction proceeded at 15°C for 25 min. The baseline separation of these derivatives was carried out on a C(8) column with methanol-tetrahydrofuran-50mM pH 6.50 HAc-NaAc buffer (55/5/40, v/v/v) as a mobile phase. Detected at the excitation and emission of 490 and 510 nm, respectively, the detection limits were obtained in the range of 0.01-0.04 nM (signal-to-noise ratio=3). The proposed method has been applied to the determination of trace aliphatic amines in viscera samples from mice without complex pretreatment or enrichment method. The recoveries ranged from 95.1% to 106.8%, depending on the samples investigated.


Soil bacteria augment Arabidopsis photosynthesis by decreasing glucose sensing and abscisic acid levels in planta.
Photosynthesis is regulated by environmental factors as well as endogenous sugar signals. Whereas light-driven sugar biosynthesis is essential for terrestrial organisms, as well as belowground microflora, whether and how soil symbionts regulate photosynthesis has yet to be reported. Here, we show that the plant growth-promoting soil bacterium Bacillus subtilis GB03 augments photosynthetic capacity by increasing photosynthetic efficiency and chlorophyll content in Arabidopsis. Mechanistic studies reveal an elevation of sugar accumulation as well as the suppression of classic glucose signaling responses, including hypocotyl elongation and seed germination, with exposure to GB03. Compared with wild-type plants, two Arabidopsis mutants defective in hexokinase-dependent sugar signaling exhibit increased photosynthetic capacity, which is not further enhanced with GB03 exposure. Overlap in sugar/ABA sensing is observed in GB03-exposed plants, with a reduction of ABA-biosynthetic transcripts as well as downstream metabolite levels in leaves. Moreover, exogenous ABA abrogates GB03-triggered increases in photosynthetic efficiency and chlorophyll content. These results demonstrate that certain rhizobacteria elevate photosynthesis through the modulation of endogenous sugar/ABA signaling, and establish a regulatory role for soil symbionts in plant acquisition of energy.


Ecological drift and competitive interactions predict unique patterns in temporal fluctuations of population size.
Recent studies have highlighted the importance of higher-order competitive interactions in stabilizing population dynamics in multi-species communities. But how does the structure of competitive hierarchies affect population dynamics and extinction processes? We tackled this important question by using spatially explicit simulations of ecological drift (10 species in a homogeneous landscape of 64 patches) in which birth rates were influenced by interspecific competition. Specifically, we examined how transitive (linear pecking orders) and intransitive (pecking orders with loops) competitive hierarchies affected extinction rates and population dynamics in simulated communities through time. In comparison to a pure neutral model, an ecological drift model including transitive competition increased extinction rates, caused synchronous density-dependent population fluctuations, and generated a white-noise distribution of population sizes. In contrast, the drift model with intransitive competitive interactions decreased extinctions rates, caused asynchronous (compensatory) density-dependent population fluctuations, and generated a brown noise distribution of population sizes. We also explored the effect on community stability of more complex patterns of competitive interactions in which pairwise competitive relationships were assigned probabilistically. These probabilistic competition models also generated density-dependent trajectories and a brown noise distribution of population sizes. However, extinction rates and the degree of population synchrony were comparable to those observed in purely neutral communities. Collectively, our results confirm that intransitive competition has a strong and stabilizing effect on local populations in species-poor communities. This effect wanes with increasing species richness. Empirical assemblages characterized by brown spectral noise, density-dependent regulation, and asynchronous (compensatory) population fluctuations may indicate a signature of intransitive competitive interactions.


[Regional variations in the incidence of manic-depressive psychoses in the Danish psychiatric hospital system 1977-1993].
The aim of the present study was to investigate geographical variations in first ever admission rates for manic-depressive psychosis during the period 1977 to 1993 at a county level. The analyses were based on data obtained from the national Danish Psychiatric Case Register comprising inpatient contacts to Danish psychiatric hospitals and psychiatric units in general hospitals. The results showed that the national first ever admission rates for manic-depressive psychosis in the age-group 15-74 years declined by 44% from 25.3 per 100,000 in 1977-1979 to 14.2 per 100,000 in 1991-1993. For the older age-group the rates were slightly increasing. Comparison of the individual counties revealed significantly geographical variations with respect to incidence level and temporal trends over the years 1977-1993. The results may reflect local differences in the provision of outpatient services and primary care facilities. However, there is also the possibility that some patients might never be treated.


Effect of the thymus on erythropoietin production in response to hypobaric hypoxia in mice.
Thymectomized, irradiated and syngeneic bone marrow-repopulated (TIR) mice, after hemopoietic recovery, were more effective in producing erythropoietin than normal controls in response to hypobaric hypoxia. Mice which were only thmectomized responded to hypoxia with more erythropoietin production than controls but less than TIR animals. It is suggested that lymphoid elements of thymic origin may produce inhibitors of the erythropoietic system. This may play a role in the anemia of lymphomas and certain other clinical syndromes.


Human Cytomegalovirus Infection Induces High Expression of Prolactin and Prolactin Receptors in Ovarian Cancer.
One of the potential biomarkers for ovarian cancer patients is high serum level of prolactin (PRL), which is a growth factor that may promote tumor cell growth. The prolactin receptor (PRLR) and human cytomegalovirus (HCMV) proteins are frequently detected in ovarian tumor tissue specimens, but the potential impact of HCMV infection on the PRL system have so far not been investigated. In this study, HCMV's effects on PRL and PRLR expression were assessed in infected ovarian cancer cells (SKOV3) by PCR and Western blot techniques. The levels of both PRL and PRLR transcripts as well as the corresponding proteins were highly increased in HCMV-infected SKOV3 cells. Tissue specimens obtained from 10 patients with ovarian cancer demonstrated high expression of PRLR, HCMV-IE, and pp65 proteins. Extensive expression of PRLR was detected in all examined ovarian tumor tissue specimens except for one from a patient who had focal expression of PRLR and this patient was HCMV-negative in her tumor. In conclusion, PRL and PRLR were induced to high levels in HCMV-infected ovarian cancer cells and PRLR expression was extensively detected in HCMV-infected ovarian tissue specimens. Highly induced PRL and PRLR by HCMV infection may be of relevance for the oncomodulatory role of this virus in ovarian cancer.


Cluster analysis to estimate the risk of preeclampsia in the high-risk Prediction and Prevention of Preeclampsia and Intrauterine Growth Restriction (PREDO) study.
Preeclampsia is divided into early-onset (delivery before 34 weeks of gestation) and late-onset (delivery at or after 34 weeks) subtypes, which may rise from different etiopathogenic backgrounds. Early-onset disease is associated with placental dysfunction. Late-onset disease develops predominantly due to metabolic disturbances, obesity, diabetes, lipid dysfunction, and inflammation, which affect endothelial function. Our aim was to use cluster analysis to investigate clinical factors predicting the onset and severity of preeclampsia in a cohort of women with known clinical risk factors. We recruited 903 pregnant women with risk factors for preeclampsia at gestational weeks 12+0-13+6. Each individual outcome diagnosis was independently verified from medical records. We applied a Bayesian clustering algorithm to classify the study participants to clusters based on their particular risk factor combination. For each cluster, we computed the risk ratio of each disease outcome, relative to the risk in the general population. The risk of preeclampsia increased exponentially with respect to the number of risk factors. Our analysis revealed 25 number of clusters. Preeclampsia in a previous pregnancy (n = 138) increased the risk of preeclampsia 8.1 fold (95% confidence interval (CI) 5.7-11.2) compared to a general population of pregnant women. Having a small for gestational age infant (n = 57) in a previous pregnancy increased the risk of early-onset preeclampsia 17.5 fold (95%CI 2.1-60.5). Cluster of those two risk factors together (n = 21) increased the risk of severe preeclampsia to 23.8-fold (95%CI 5.1-60.6), intermediate onset (delivery between 34+0-36+6 weeks of gestation) to 25.1-fold (95%CI 3.1-79.9) and preterm preeclampsia (delivery before 37+0 weeks of gestation) to 16.4-fold (95%CI 2.0-52.4). Body mass index over 30 kg/m2 (n = 228) as a sole risk factor increased the risk of preeclampsia to 2.1-fold (95%CI 1.1-3.6). Together with preeclampsia in an earlier pregnancy the risk increased to 11.4 (95%CI 4.5-20.9). Chronic hypertension (n = 60) increased the risk of preeclampsia 5.3-fold (95%CI 2.4-9.8), of severe preeclampsia 22.2-fold (95%CI 9.9-41.0), and risk of early-onset preeclampsia 16.7-fold (95%CI 2.0-57.6). If a woman had chronic hypertension combined with obesity, gestational diabetes and earlier preeclampsia, the risk of term preeclampsia increased 4.8-fold (95%CI 0.1-21.7). Women with type 1 diabetes mellitus had a high risk of all subgroups of preeclampsia. The risk of preeclampsia increases exponentially with respect to the number of risk factors. Early-onset preeclampsia and severe preeclampsia have different risk profile from term preeclampsia.


The importance of autumn rainfall and sentinel flock location to understanding the epidemiology of St. Louis encephalitis virus in Indian River County, Florida.
Culex nigripalpus adults were collected from sentinel chickens at 4 sites in Indian River Co. (IRC), Florida. Chickens located in well drained, open habitats attracted fewer Cx. nigripalpus than did those located in a moist, forested area, but the number of mosquitoes collected in the open habitats increased significantly with heavy autumn rainfall. All St. Louis encephalitis virus (SLE) seroconversions (11/674) during a 7-year period were from chickens located in open, well drained habitats. A comparison of monthly rainfall during years of known SLE activity in IRC with the 22-year average showed that virus activity was significantly associated with years in which rainfall was unusually low in either September or October. Sentinel chickens in areas with large vector populations may not be the most reliable indicators of local SLE activity and autumn rainfall in September and October may influence yearly SLE patterns in southern Florida.


Investigation of pH Influence on Skin Permeation Behavior of Weak Acids Using Nonsteroidal Anti-Inflammatory Drugs.
As a continuing effort to understand the skin permeation behavior of weak acids and bases, the objectives of the present study were to evaluate skin permeation of nonsteroidal anti-inflammatory drugs (NSAIDs) under the influence of pH, investigate the mechanism of pH effect, and examine a previous hypothesis that the effective skin pH for drug permeation is different from donor solution pH. In vitro permeability experiments were performed in side-by-side diffusion cells with diclofenac, ibuprofen, flurbiprofen, ketoprofen, and naproxen and human skin. The donor solution pH significantly affected skin permeation of NSAIDs, whereas no effect of the receiver pH was observed. Similar to previous observations, the apparent permeability coefficient versus donor solution pH relationships deviated from the predictions (fractions of unionized NSAIDs) according to the acid/base theory. The influences of the viable epidermis barrier, polar pathway transport, ion permeation across skin, and effective skin pH were investigated. The effective pH values for skin permeation determined using the NSAIDs (weak acids) in this study were different from those obtained previously with a weak base at the same donor solution pH conditions, suggesting that the observed permeability-pH relationships could not be explained solely by possible pH differences between skin and donor solution. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:3459-3470, 2015.


Monobloc Frontofacial or Le Fort III Distraction Osteogenesis in Syndromic Craniosynostosis: Three-Dimensional Evaluation of Treatment Outcome and the Need for Central Distraction.
The objectives of this study were to investigate the treatment effect, morphology, and volumetric outcomes of monobloc frontofacial or Le Fort III distraction osteogenesis in syndromic craniosynostosis by 3-dimensional evaluation. Nine consecutive patients underwent monobloc frontofacial or Le Fort III distraction during 2003 to 2012 were included and evaluated. The patient's evaluation is a minimum of a 4-year follow-up. Pretreatment, post-treatment advancement, and relapse were quantified. The changes in intracranial volume, upper airway volume, globe protrusion, advancement at bilateral malar eminence, and central face were calculated from computerized tomography before and after treatment. After distraction, the intracranial volume was increased by 16.4% in average, and the upper airway volume increased by 64.1%. Orbital protrusion improved by 9.9 mm in the left eye and 10.5 mm in the right eye in comparison to the preoperative status. Bilateral malar eminences advancement was greater than the dorsum advancement by 7.1 mm. The external distraction osteogenesis device caused significant technical difficulties with advancement of the midface in growing bones. The central midface did not advance as hoped for. This finding may suggest a need for modification in the distraction device in order to improve the central facial distraction vector.


A substance use prevention framework: considering the social context for African American girls.
Shifting patterns of substance use (that is, early initiation, increased marijuana use, narrowing differences in gender use) and the disproportionate socioeconomic obstacles that are related to substance use among ethnically diverse adolescent females create the need to develop ethnic and gender-specific substance use prevention frameworks. This article describes and applies a substance use prevention framework to African American females. Gender socialization and self-efficacy are presented as key concepts, along with the assertion that every substance use prevention framework should examine the influences of specific societal factors (such as racism, sexism, classism, and ageism) on substance use. Rationale and guidelines for designing ethnically sensitive and gender-specific research projects and intervention programs regarding substance use prevention are offered. Public health nurses (PHNs) are uniquely positioned to use this framework in their work with African American adolescent girls, specifically, and in general with other ethnically diverse groups.


Forward fiber Fourier transform spectrometer modeling and design with PZT phase modulation real-time compensation.
Piezoelectric ceramic transducers (PZTs) are often applied in all-fiber Fourier transform spectrometers (FFTSs) to realize phase modulation. Combining a PZT and optical fiber features, a theoretical FFTS modeling is established. We systematically deduced the main causes of spectral errors and the factors of the instrumental resolution, then designed a new FFTS system and provided real-time compensation methods for spectral errors. We creatively employed two Mach-Zehnder interferometers by winding the sensing arms of the two on the same PZT cylinder to realize the dual optical path experiencing the same and simultaneous phase modulation. A processing circuit is designed to achieve selective sampling of the test interferogram, avoiding the spectral artifacts generated by the PZT phase discontinuities. The narrow line width of the reference spectrum is obtained at the same time for real-time calibration of spectral shift resulting from dispersion and other phase errors. By the comparison of the original interferogram and spectrum with those after compensation, the experiments validated that the system design could effectively compensate for the spectral errors caused by PZT and optical fiber physical limitations. Finally, the improved ideas are discussed if the FFTS prototype is to be applied to test a real-time gas spectrum.


Cardiovascular Benefits of Fish-Oil Supplementation Against Fine Particulate Air Pollution in China.
Few studies have evaluated the health benefits of omega-3 fatty acid supplementation against fine particulate matter (aerodynamic diameter <2.5 μm [PM2.5]) exposure in highly polluted areas. The authors sought to evaluate whether dietary fish-oil supplementation protects cardiovascular health against PM2.5 exposure in China. This is a randomized, double-blinded, and placebo-controlled trial among 65 healthy college students in Shanghai, China. Participants were randomly assigned to either the placebo group or the intervention group with dietary fish-oil supplementation of 2.5 g/day from September 2017 to January 2018, and received 4 rounds of health examinations in the last 2 months of treatments. Fixed-site PM2.5 concentrations on campus were measured in real time. The authors measured blood pressure and 18 biomarkers of systematic inflammation, coagulation, endothelial function, oxidative stress, antioxidant activity, cardiometabolism, and neuroendocrine stress response. Acute effects of PM2.5 on these outcomes were evaluated within each group using linear mixed-effect models. The average PM2.5 level was 38 μg/m3 during the study period. Compared with the placebo group, the fish-oil group showed relatively stable levels of most biomarkers in response to changes in PM2.5 exposure. Between-group differences associated with PM2.5 exposure varied by biomarkers and by lags of exposure. The authors observed beneficial effects of fish-oil supplementation on 5 biomarkers of blood inflammation, coagulation, endothelial function, oxidative stress, and neuroendocrine stress response in the fish-oil group at a false discovery rate of <0.05. This trial shows that omega-3 fatty acid supplementation is associated with short-term subclinical cardiovascular benefits against PM2.5 exposure among healthy young adults in China. (Effect of Dietary Supplemental Fish Oil in Alleviating Health Hazards Associated With Air Pollution; NCT03255187).


The spread of the cult of Asclepius in the context of the Roman army benefited from the presence of physicians: A spatial proximity analysis.
The article applies a GIS based approach to the study of the spread of the cult of Asclepius, the Greco-Roman healing god, during the Roman period. It explores the role of soldiers and physicians in the spatial dissemination of the cult along the transportation network of Roman roads in the border provinces of Britannia, Germania Superior and Inferior, Raetia, Noricum, Pannonia Superior and Inferior, Moesia Superior and Inferior, and Dacia. These provinces were selected as a suitable area for quantitative GIS exploration because they were all on the outer border of the Roman Empire, had a significant military presence, and there is a representative amount of inscriptions attested that can be used as proxies for the spatial occurrence of the three measured variables: the cult of Asclepius, Roman soldiers, and Roman physicians. After establishing by means of spatial proximity analysis that the cult of Asclepius occurred frequently in the context of the Roman army, the article proposes and quantitatively evaluates a more specific hypothesis; i.e., that the spatial occurrences of Roman physicians in inscriptions are a relevant predictor for the spatial occurrences of the worship of Asclepius in the environment of the Roman army because of the shared focus between physicians and the cult of Asclepius-health and medicine. The highly significant results of the statistical analysis reveal a positive trend in the spatial relationships between Roman physicians and the worship of Asclepius in the context of the Roman army in the majority of provinces of interest, thus supporting the proposed hypothesis. The results presented in the article demonstrate the potential of the GIS approach in testing assumptions produced by traditional scholarship and in nuancing our understanding of a specific process of cultural spread.


Efficacy of imidacloprid 10%/moxidectin 2.5% spot on (Advocate®, Advantage Multi®) and doxycycline for the treatment of natural Dirofilaria immitis infections in dogs.
Heartworm infection (also known as dirofilariosis due to Dirofilaria immitis) in dogs causes chronic pulmonary disease that, if left untreated, can lead to right-side congestive heart failure. Currently, the only registered drug for adulticide therapy in dogs with heartworm disease (HWD) is melarsomine dihydrochloride. The recent targeting of the bacterial endosymbiont Wolbachia, through antibiotic therapy of the infected host, has offered an interesting alternative for the treatment of HWD. Recent reports of the adulticide activity of an ivermectin/doxycycline combination protocol has lead the American Heartworm Society (AHS) to include in its guidelines that, in cases where arsenical therapy is not possible or is contraindicated, a monthly heartworm preventive along with doxycycline for a 4-week period might be considered. In the present study, 20 dogs with confirmed natural D. immitis infection were included following owner consent. Fourteen dogs were treated with a topical formulation containing 10% w/v imidacloprid and 2.5% w/v moxidectin (Advocate®, Advantage Multi®, Bayer), monthly for nine months, associated to doxycycline (10 mg/kg/BID) for the first 30 days. Six dogs were treated with melarsomine (Immiticide®, Merial) (2.5 mg/kg) at enrollment, followed one month later by two injections 24 h apart. The presence of circulating antigens and the number of microfilariae (mf) were evaluated at the moment of enrollment and then at 1, 2, 3, 4, 5, 6, 7, 8, 12, 18, 24 months post enrollment. Echocardiogram and radiographs were performed at month 0, 6, 12, 18, 24. Monthly moxidectin combined with 30 days of doxycycline eliminated circulating microfilariae within one month, thus breaking the transmission cycle very quickly. Furthermore, dogs treated with the combination protocol started to become negative for circulating antigens at 4 months from the beginning of treatment and all except one were antigen negative at 9 months. All dogs treated with melarsomine were antigen negative by 5 months from the beginning of the treatment. No dogs showed worsening of pulmonary patterns or criteria indicative of pulmonary hypertension 12 to 24 months after. For the criteria mf concentration, antigen concentration, radiography and echocardiography at 12, 18 and 24 months the non-inferiority for the moxidectin group could be proven for a non-inferiority margin of 15% for the rate difference. Dogs treated with moxidectin and doxycycline became negative for microfilariae and antigens sooner when compared to melarsomine in the present study and to dogs treated with doxycycline combined with ivermectin in studies previously published.


Two components of the chloroplast protein import apparatus, IAP86 and IAP75, interact with the transit sequence during the recognition and translocation of precursor proteins at the outer envelope.
The interactions of precursor proteins with components of the chloroplast envelope were investigated during the early stages of protein import using a chemical cross-linking strategy. In the absence of energy, two components of the outer envelope import machinery, IAP86 and IAP75, cross-linked to the transit sequence of the precursor to the small subunit of ribulose-1, 5-bisphosphate carboxylase (pS) in a precursor binding assay. In the presence of concentrations of ATP or GTP that support maximal precursor binding to the envelope, cross-linking to the transit sequence occurred predominantly with IAP75 and a previously unidentified 21-kD polypeptide of the inner membrane, indicating that the transit sequence had inserted across the outer membrane. Cross-linking of envelope components to sequences in the mature portion of a second precursor, preferredoxin, was detected in the presence of ATP or GTP, suggesting that sequences distant from the transit sequence were brought into the vicinity of the outer membrane under these conditions. IAP75 and a third import component, IAP34, were coimmunoprecipitated with IAP86 antibodies from solubilized envelope membranes, indicating that these three proteins form a stable complex in the outer membrane. On the basis of these observations, we propose that IAP86 and IAP75 act as components of a multisubunit complex to mediate energy-independent recognition of the transit sequence and subsequent nucleoside triphosphate-induced insertion of the transit sequence across the outer membrane.


Characterization of patients with angioscopically-detected in-stent mural thrombi – genetics of clopidogrel responsiveness and generations of drug-eluting stents.
The loss-of-function genotype of cytochrome P450 2C19 (CYP2C19) has been proposed as a risk factor for stent thrombosis in patients with drug-eluting stent implantation. The aim of this study was to clarify the clinical features of patients with angioscopically-detected in-stent mural thrombi (ISMT). Enrolled were 100 stented segments in 55 patients with stable angina (20 bare-metal stents; 39 Cypher sirolimus-eluting stents [SES]; 26 Endeavor zotarolimus-eluting stents [ZES]; 13 Xience V everolimus-eluting stents; and 2 Nobori biolimus-eluting stents). Dual antiplatelet therapy (100 mg aspirin+75 mg clopidogrel once daily) had been continued since stenting. A poor metabolizer (PM) of clopidogrel was defined as a homozygote of CYP2C19 loss-of-function alleles. Coronary angioscopy revealed ISMT in 6 patients (5 SES, 1 ZES). Between the ISMT group and control group (n=49), there were no significant differences with regards to the VerifyNow P2Y12platelet function assay or in-stent endothelial coverage grade. Exact logistic regression analyses with stepwise forward selection at a significance level of 0.10 were performed to reveal predictive variables for ISMT (respectively: odds ratio, 95% confidence interval, P value: CYP2C19 PM genotype (3.28, 0.88-24.80, 0.09), SES implantation (3.37, 0.90-28.09, 0.08), and presence of yellow plaque (3.69, 1.14-25.70, 0.02). Patients with ISMT were characterized by SES implantation, poor clopidogrel metabolism, and in-stent yellow plaque.


Scoliosis and pregnancy.
The effects of pregnancy on patients who have idiopathic scoliosis were investigated in terms of increased risk of progression of the curve. The charts, radiographs, and other pertinent data on 355 affected women who had reached skeletal maturity (Risser Grade 4) before 1975 were reviewed and analyzed. One hundred and seventy-five patients had had at least one pregnancy each (Group A) and 180 patients had never been pregnant (Group B). The groups were comparable with regard to the treatments that they had received. After skeletal maturity was reached, the curve progressed more than 5 degrees in 25 per cent and more than 10 degrees in 10 per cent of the patients in each group. The age of the patient at the time of the first pregnancy did not influence the risk of progression, and the stability of the curve before pregnancy did not decrease the risk of its progression during pregnancy. In patients who had had a spinal fusion, progression in the unfused portion of the spine was negligible in both Group A and Group B. The presence of a pseudarthrosis did not result in progression of the curve during pregnancy. The effects of scoliosis on pregnancy and delivery were evaluated in the 175 women in Group A. No specific problems that were directly related to the scoliosis were noted except for four patients, in whom delivery posed difficulties. The incidence of cesarean section was one-half of the national average, and no sections were directly related to the mother's scoliosis.(ABSTRACT TRUNCATED AT 250 WORDS)


Nuclear carbonic anhydrase 6B associates with PRMT5 to epigenetically promote IL-12 expression in innate response.
Interleukin-12 (IL-12) is critical for induction of protective immunity against intracellular bacterial infection. However, the mechanisms for efficient induction of IL-12 in innate response remain poorly understood. Here we report that the B type of carbonic anhydrase 6 (Car6-b, which encoded CA-VI B) is essential for host defense against Listeria monocytogenes (LM) infection by epigenetically promoting IL-12 expression independent of its carbonic anhydrase activity. Deficiency of Car6-b attenuated IL-12 production upon LM infection both in vitro and in vivo. Car6-/- mice were more susceptible to LM infection with less production of IL-12. Mechanistically, the nuclear localized CA-VI B selectively promotes IL-12 expression by interaction with protein arginine N-methyltransferase 5 (PRMT5), which reduces symmetric dimethylation of histone H3 arginine 8 modification (H3R8me2s) at Il12 promoters to facilitate chromatin accessibility, selectively enhancing c-Rel binding to the Il12b promoter. Our findings add insights to the epigenetic regulation of IL-12 induction in innate immunity.


One-pot construction of nitrogen-rich polymeric ionic porous networks for effective CO2 capture and fixation.
Facile preparation of ionic porous networks (IPNs) with large and permanent porosity is highly desirable for CO2 capture and transformation but remains a challenge. Here we report a one-pot base-mediated construction of nitrogen-rich IPNs through a combination of nucleophilic substitution and quaternisation chemistry from H-imidazole. This strategy, as proven by the model reactions of 1H-imidazole or 1-methyl-1H-imidazole with cyanuric chloride, allows for fine regulation of porosity and physicochemical properties, leading to nitrogen-rich IPNs featuring abundant ionic units and radicals. The as-prepared networks, termed IPN-CSUs, efficiently capture CO2 (80.1 cc g-1 at 273 K/1 bar) with an ideal CO2/N2 selectivity of 139.7. They can also effectively catalyse the cycloaddition reaction between CO2 and epoxides with high yields of up to 99% under mild conditions (0.1 MPa, 298 K), suggesting their possible applications in the fields of both selective molecular separation and conversion. Unlike the previously known strategies generally involving single coupling chemistry, our strategy combining two coupling routes in one pot appears to be unique and potentially applicable to other building blocks.


Mass flux from a non-aqueous phase liquid pool considering spontaneous expansion of a discontinuous gas phase.
The partitioning of non-aqueous phase liquid (NAPL) compounds to a discontinuous gas phase results in the repeated spontaneous expansion, snap-off, and vertical mobilization of the gas phase. This mechanism has the potential to significantly affect the mass transfer processes that control the dissolution of NAPL pools by increasing the vertical transport of NAPL mass and increasing the total mass transfer rate from the surface of the pool. The extent to which this mechanism affects mass transfer from a NAPL pool depends on the rate of expansion and the mass of NAPL compound in the gas phase. This study used well-controlled bench-scale experiments under no-flow conditions to quantify for the first time the expansion of a discontinuous gas phase in the presence of NAPL. Air bubbles placed in glass vials containing NAPL increased significantly in volume, from a radius of 1.0 mm to 2.0 mm over 215 days in the presence of tetrachloroethene (PCE), and from a radius of 1.2 mm to 2.3 mm over 22 days in the presence of trans-1,2-dichloroethene (tDCE). A one-dimensional mass transfer model, fit to the experimental data, showed that this expansion could result in a mass flux from the NAPL pool that was similar in magnitude to the mass flux expected for the dissolution of a NAPL pool in a two-fluid (NAPL and water) system. Conditions favouring the significant effect of a discontinuous gas phase on mass transfer were identified as groundwater velocities less than approximately 0.01 m/day, and a gas phase that covers greater than approximately 10% of the pool surface area and is located within approximately 0.01 m of the pool surface. Under these conditions the mass transfer via a discontinuous gas phase is expected to affect, for example, efforts to locate NAPL source zones using aqueous concentration data, and predict the lifetime and risk associated with NAPL source zones in a way that is not currently included in the common conceptual models used to assess NAPL-contaminated sites.


Direct electrochemical measurements inside a 2000 angstrom thick polymer film by scanning electrochemical microscopy.
An extremely small, conically shaped Pt microelectrode tip (with a radius of 30 nanometers) and the precise positioning capabilities of the scanning electrochemical microscope were used to penetrate a thin (200 nanometers) polymer film and obtain directly the standard potential and kinetic parameters of an electrode reaction within the film. The thickness of the film was determined while it was immersed in and swollen by an electrolyte solution. The film studied was the perfluorosulfonate Nafion containing Os(bpy)(3)(2+) (bpy, 2,2'-bipyridine) cast on an indium tin oxide surface. The steady-state response at the ultramicroelectrode allowed direct determination of the rate constant for heterogeneous electron transfer K(o) and the diffusion coefficient D without complications caused by transport in the liquid phase, charge exchange at the liquid-polymer interface, and resistive drop.


Adaptive multiscale molecular dynamics of macromolecular fluids.
Until now, adaptive atomistic-coarse-grain (A/CG) molecular dynamics simulations have had very limited applicability because the on-the-fly CG → A transformation is problematic for all but those molecules whose CG representation consists of a single particle. Here, we solve this problem by combining a transitional healing region with a rotational dynamics of rigid atomistic fragments in the CG region. Error control is obtained by analysis of the A ↔ CG energy flow. We illustrate the method with adaptive multiscale simulations of liquid hexane and of a dilute polymer solution in a theta solvent.


Computational approaches to studying methylated H4K20 recognition by DNA repair factor 53BP1.
Histone lysine methylation regulates the recruitment of mammalian DNA repair factor 53BP1 to the histone H4 lysine 20 (H4K20), through specific recognition of the tandem Tudor domain of 53BP1. The di- and mono-methylated H4K20 bind to 53BP1 with high affinity, but the non- and tri-methylated H4K20 do not. Here, we develop a new approach to carry out computational study to unravel the binding mechanism of methylated H4K20 by 53BP1 and to compute relative binding affinities of different methylations of H4K20 by 53BP1. First, hot spots in 53BP1 were predicted by computational alanine scanning and aromatic cages formed by W1495, Y1500, Y1502, and Y1523 are found to provide the dominant binding to di- and mono-methylated H4K20 in addition to D1521. Secondly, a de-methylation method is proposed to predict relative binding free energies between 53BP1 and different methylated states of H4K20. Finally, the tri-methylated and non-methylated H4K20/53BP1 complexes are found to be dynamically unstable, explaining the experimental finding that neither can bind to 53BP1. The present work provides an important theoretical basis for our understanding of histone methylations of H4K20 and their recognition mechanism by DNA repair factor 53BP1.


[Study on HPLC fingerprint of Taxus chinensis var. mairei].
To establish the HPLC fingerprint of Taxus chinensis var. mairei collected from different parts during different seasons and provide scientific basis for its comprehensive utilization. Supercritical CO2 extraction was used to extract the effective fraction, HPLC method to establish the fingerprint and similarity evaluation software to analyze the fingerprint chromatogram. 12 batches of Taxus chinensis var. mairei medicinal materials from different parts collected in different seasons were analyzed and HPLC fingerprint of Taxus chinensis var. mairei was established. The HPLC fingerprint can be used to evaluate the quality of Taxus chinensis var. mairei medicinal materials.


[Prevention of upper limbs diseases in workers subjected to occupational hazards].
Study covered 120 personal computer users, 98 jewelry female polishers, 64 metallic ships body assemblers. Objective investigations revealed joints pain on palpation, joints function disorders, hypoesthesia of finger tips. After prevention measures based on low-intensity laser rays, the joints functions recovered, hands edema disappeared, hypoesthesia cleared.


Regulation of hepatitis C virus replication by nuclear translocation of nonstructural 5A protein and transcriptional activation of host genes.
Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) is involved in regulating viral replication through its direct interaction with the HCV RNA-dependent RNA polymerase. NS5A also alters infected cell metabolism through complex interactions with numerous host cell proteins. NS5A has furthermore been suggested to act as a transcriptional activator, although the impact on viral replication is unclear. To study this, HCV NS5A variants were amplified from hepatic tissue from an HCV-infected patient, and their abilities to activate gene transcription were analyzed in a single-hybrid yeast (Saccharomyces cerevisiae) model. Different variants isolated from the same patient displayed different transactivational activities. When these variants were inserted into the HCV subgenomic replicon system, they demonstrated various levels of RNA replication, which correlated with their transactivational activities. We showed that the C-terminal fragment of NS5A was localized to the nucleus and that a functional NS5A nuclear localization signal and cellular caspase activity were required for this process. Furthermore, nuclear localization of NS5A was necessary for viral replication. Finally, we demonstrate that nuclear NS5A binds to host cell promoters of several genes previously identified as important for efficient HCV RNA replication, inducing their transcription. Taken together, these results demonstrate a new mechanism by which HCV modulates its cellular environment, thereby enhancing viral replication.


Release and complex formation of soluble VEGFR-1 from endothelial cells and biological fluids.
One of the key molecules promoting angiogenesis is the endothelial cell-specific mitogen, vascular endothelial growth factor (VEGF or VEGF-A), which acts through two high-affinity receptor tyrosine kinases (VEGFR), VEGFR-1 (or Flt-1) and VEGFR-2 (or KDR/Flk-1). It was shown before that a soluble variant of VEGFR-1 (sVEGFR-1) can be generated by differential splicing of the flt-1 mRNA. This soluble receptor is an antagonist to VEGF action, reducing the level of free, active VEGF-A, and therefore, plays a pivotal role in the generation of vascular diseases like pre-eclampsia or intra-uterine growth retardation. Here we show that sVEGFR-1 is produced by cultured human microvascular and macrovascular endothelial cells and a human melanoma cell line. The soluble receptor is mainly complexed with ligands; only 5-10% remains detectable as free, uncomplexed receptor protein. Furthermore, we show the time course of total and free sVEGFR-1 release together with its putative ligands, VEGF-A and placenta growth factor (PIGF), from macrovascular endothelial cells. The release of sVEGFR-1 was quantitatively measured in two different ELISA types. The release of sVEGFR-1 was strongly enhanced by phorbol-ester (PMA); the cells produced up to 22 ng/ml of sVEGFR-1 after 48 hours. The expression of VEGF-A and PIGF was moderately influenced by PMA. We also show a hypoxia-induced increase of sVEGFR-1 expression in cells cultured from placenta, a tissue that has a high flt-1 gene expression. Moreover, we demonstrate that sVEGFR-1 in amniotic fluids acts as a sink for exogenous VEGF165 and PIGF-2. Here, for the first time, to what extent recombinant ligands have to be added to compensate for the sink function of amniotic fluids was analyzed. In conclusion, human endothelial cells produce high levels of sVEGFR-1, which influences the availability of VEGF-A or related ligands. Therefore, sVEGFR-1 may reduce the ligand binding to transmembrane receptors and interfere with their signal transduction.


Production of fungal fructosyl amino acid oxidase useful for diabetic diagnosis in the peroxisome of Candida boidinii.
A high-level production of fructosyl amino acid oxidase (FAOD), whose production was toxic in Escherichia coli, was investigated through attempts to utilize the peroxisome of Candida boidinii as the place for protein accumulation. The alcohol oxidase-depleted strain (strain aod1Delta) produced FAOD at a four to five times higher level than the wild type strain in terms of protein amount and enzyme activity, although the transcriptional level was similar. As a result of this study, we could improve FAOD productivity approximately 47-fold from the original transformant, and FAOD accumulated within membrane-bound peroxisomes up to 18% of the total soluble proteins.


Target search on a dynamic DNA molecule.
We study a protein-DNA target search model with explicit DNA dynamics applicable to in vitro experiments. We show that the DNA dynamics plays a crucial role for the effectiveness of protein "jumps" between sites distant along the DNA contour but close in three-dimensional space. A strongly binding protein that searches by one-dimensional sliding and jumping alone explores the search space less redundantly when the DNA dynamics is fast on the time scale of protein jumps than in the opposite "frozen DNA" limit. We characterize the crossover between these limits using simulations and scaling theory. We also rationalize the slow exploration in the frozen limit as a subtle interplay between long jumps and long trapping times of the protein in "islands" within random DNA configurations in solution.


Calcium-regulatory mechanisms. Functional classification using skinned fibers.
The primary purpose of this study was to determine whether various agents (adenosine 3-thiotriphosphate [ATP gamma S], trifluoperazine [TFP], troponin I, the catalytic subunit of the cyclic adenosine 3',5'-monophosphate dependent protein kinase [C-subunit], and calmodulin [CaM]) could be used to classify skinned fiber types, and then to determine whether the proposed mechanisms for Ca2+ regulation were consistent with the results. Agents (ATP gamma S, TFP, C-subunit, CaM) expected to alter a light chain kinase-phosphatase system strongly affect the Ca2+-activated tension in skinned gizzard smooth muscle fibers, whereas these agents have no effect on skinned mammalian striated and scallop adductor fibers. Troponin I, which is known to bind strongly to troponin C and CaM, inhibits Ca2+ activation of skinned mammalian striated and gizzard fibers but not scallop adductor muscle. The results in different types of skinned fibers are consistent with proposed mechanisms for Ca2+ regulation.


Ongoing Computed Tomography Appraisal of Intestinal Perforation Due to an Ingested Foreign Body.
BACKGROUND Diagnosis and management of accidental or intentional ingestion of foreign bodies is a common problem at in emergency departments. This condition is generally observed in patients with limited consciousness or attention, such as children, elders, or psychiatric patients. Here, we report a case of intestinal perforation caused by ingestion of a foreign body that occurred during the performance of a contrast-enhanced CT scan. CASE REPORT A 73-year-old diabetic woman was admitted to the emergency room of our hospital with postprandial abdominal discomfort, nausea, and vomiting. Under the suspicion of bowel ischemia, the patient underwent a contrast-enhanced CT scan. A thickened ileal loop with an endoluminal bone-density foreign body was detected. The following contrast-enhanced acquisitions additionally showed air bubbles adjacent to the loop, as the sign of an intestinal perforation that occurred between the basal and the contrast-enhanced acquisitions. CONCLUSIONS Caution should be always exercised in patients with suspected gastrointestinal perforation, especially if caused by ingested foreign bodies. A high degree of suspicion and a CT scan may prevent delays in the diagnosis and clinical management of these patients.


Psychedelics for treatment resistant depression: are they game changers?
A new era of treatment for adults with treatment-resistant depression (TRD), which involves psychedelic substances, is dawning. Emerging evidence indicates that psychedelics can exert antidepressant effects through multiple neurobiological and psychological mechanisms. However, it remains to be seen if these new treatments will revolutionize the treatment of TRD. The present review focuses on the efficacy of serotoninergic psychedelics psilocybin, lysergic acid diethylamide (LSD), N,N-dimethyltryptamine (DMT), ayahuasca, 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) and mescaline (3,4,5-trimethoxyphenethylamine), as well as 3,4-methylenedioxymethamphetamine (MDMA), for TRD. A systematic search was conducted for psilocybin in TRD as emerging trials had not yet been subject to review. A narrative review summarized findings on other psychedelics. Psychedelic therapy has created a paradigm shift in the treatment of TRD, as it can maximize therapeutic benefits and minimize potential risks. Psilocybin holds promise as a potential game-changer in the treatment of TRD, with initial evidence suggesting a rapid antidepressant effect sustained for some responders for at least 3 months. Nevertheless, further adequately powered, double-blind, comparator-controlled trials are required to explore and clarify the mechanisms of action and long-term effects of psychedelics in TRD. Psychedelics also hold promise for other psychiatric conditions, such as bipolar depression and post-traumatic stress disorder.


Identification of an alpha3-fucosyltransferase and a novel alpha2-fucosyltransferase activity in cercariae of the schistosome Trichobilharzia ocellata: biosynthesis of the Fucalpha1-->2Fucalpha1-->3[Gal(NAc)beta1-->4]GlcNAc sequence.
Fucose is a major constituent of the protein- and lipid-linked glycans of the various life-cycle stages of schistosomes. These fucosylated glycans are highly antigenic and seem to play a role in the pathology of schistosomiasis. In this article we describe the identification and characterization of two fucosyltransferases (FucTs) in cercariae of the avian schistosome Trichobilharzia ocellata, a GDP-Fuc:[Galbeta1-->4]GlcNAcbeta-R alpha1-->3-FucT and a novel GDP-Fuc:Fucalpha-R alpha1-->2-FucT. Triton X-100 extracts of cercariae were assayed for FucT activity using a variety of acceptor substrates. Type 1 chain (Galbeta1-->3GlcNAc) based compounds were poor acceptors, whereas those based on a type 2 chain (Galbeta1-->4GlcNAc), whether alpha2'-fucosylated, alpha3'-sialylated, or unsubstituted, and whether present as oligosaccharide or contained in a glycopeptide or glycoprotein, all served as acceptor substrates. In this respect the schistosomal alpha3-FucT resembles human FucT V and VI rather than other known FucTs. N-ethylmaleimide, an inhibitor of several human FucTs, had no effect on the activity of the schistosomal alpha3-FucT, whereas GDP-beta-S was strongly inhibitory. Large scale incubations were carried out with Galbeta1-->4GlcNAc, GalNAcbeta1-->4GlcNAcbeta-O -(CH2)8COOCH3 and Fucalpha1-->3GlcNAcbeta1-->2Man as acceptor substrates and the products of the incubations were isolated using a sequence of chromatographic techniques. By methylation analysis and 2D-TOCSY and ROESY1H-NMR spectroscopy the products formed were shown to be Galbeta1-->4[Fucalpha1-->2Fucalpha1-->3]GlcNAc, GalNAcbeta1-->4[Fucalpha1-->2Fucalpha1-->3]GlcNAcbe ta-O-(CH2)8COOCH3, and Fucalpha1-->2Fucalpha1-->3GlcNAcbeta1-->2Man, respectively. It is concluded that the alpha2-FucT and alpha3-FucT are involved in the biosynthesis of the (oligomeric) Lewisx sequences and the Fucalpha1-->2Fucalpha1-->3GlcNAc structural element that have been described on schistosomal glycoconjugates.


Palpable splenomegaly in children with haemoglobin SC disease: haematological and clinical manifestations.
This study aimed to investigate the prevalence of palpable splenomegaly in children with haemoglobin SC (Hb SC) disease, and to determine the haematological and clinical manifestations of splenomegaly in this patient population. We performed a retrospective chart review of 100 patients with Hb SC over 2 years of age followed by the Duke University Paediatric Sickle Cell Program with serial physical examinations and laboratory measurements. Palpable splenomegaly was present in 34% of patients and was more common in males (P = 0.029). Children with splenomegaly had a significantly lower average haemoglobin concentration (10.3 vs. 10.8 g/dl, P = 0.011) and lower platelet count (237 vs. 314 x 109/l, P < 0.001) than those without splenomegaly. Children with measurements both before and after the onset of splenomegaly had a significant decrease in the platelet count (279 vs. 216 x 109/l, P < 0.001) and white blood cell count (9.1 vs. 7.9 x 109/l, P = 0.04) after splenomegaly was identified. Clinical complications included acute splenic sequestration in 12% of children (median age 5.4 years), and hypersplenism with chronic thrombocytopenia in another 10% of patients (median age 10.6 years). Splenomegaly is a common physical finding in children with Hb SC disease and is often associated with mild cytopenias. Clinical complications of splenomegaly include acute splenic sequestration in younger patients and hypersplenism with chronic thrombocytopenia in older children.


Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain.
The Blue Light Using Flavin (BLUF) domain proteins are an important family of photoreceptors controlling a range of responses in a wide variety of organisms. The details of the primary photochemical mechanism, by which light absorption in the isoalloxazine ring of the flavin is converted into a structure change to form the signalling state of the protein, is unresolved. In this work we apply ultrafast time resolved infra-red (TRIR) spectroscopy to investigate the primary photophysics of the BLUF domain of the protein AppA (AppABLUF) a light activated antirepressor. Here a number of mutations at Y21 and W104 in AppABLUF are investigated. The Y21 mutants are known to be photoinactive, while W104 mutants show the characteristic spectral red-shift associated with BLUF domain activity. Using TRIR we observed separately the decay of the excited state and the recovery of the ground state. In both cases the kinetics are found to be non-single exponential for all the proteins studied, suggesting a range of ground state structures. In the Y21 mutants an intermediate state was also observed, assigned to formation of the radical of the isoalloxazine (flavin) ring. The electron donor is the W104 residue. In contrast, no radical intermediates were detected in the studies of the photoactive dark adapted proteins, dAppABLUF and the dW104 mutants, suggesting a structure change in the Y21 mutants which favours W104 to isoalloxazine electron transfer. In contrast, in the light adapted form of the proteins (lAppABLUF, lW104) a radical intermediate was detected and the kinetics were greatly accelerated. In this case the electron donor was Y21 and major structural changes are associated with the enhanced quenching. In AppABLUF and the seven mutants studied radical intermediates are readily observed by TRIR spectroscopy, but there is no correlation with photoactivity. This suggests that if a charge separated state has a role in the BLUF photocycle it is only as a very short lived intermediate.


Clinical relevance assessment of animal preclinical research (RAA) tool: development and explanation.
Only a small proportion of preclinical research (research performed in animal models prior to clinical trials in humans) translates into clinical benefit in humans. Possible reasons for the lack of translation of the results observed in preclinical research into human clinical benefit include the design, conduct, and reporting of preclinical studies. There is currently no formal domain-based assessment of the clinical relevance of preclinical research. To address this issue, we have developed a tool for the assessment of the clinical relevance of preclinical studies, with the intention of assessing the likelihood that therapeutic preclinical findings can be translated into improvement in the management of human diseases. We searched the EQUATOR network for guidelines that describe the design, conduct, and reporting of preclinical research. We searched the references of these guidelines to identify further relevant publications and developed a set of domains and signalling questions. We then conducted a modified Delphi-consensus to refine and develop the tool. The Delphi panel members included specialists in evidence-based (preclinical) medicine specialists, methodologists, preclinical animal researchers, a veterinarian, and clinical researchers. A total of 20 Delphi-panel members completed the first round and 17 members from five countries completed all three rounds. This tool has eight domains (construct validity, external validity, risk of bias, experimental design and data analysis plan, reproducibility and replicability of methods and results in the same model, research integrity, and research transparency) and a total of 28 signalling questions and provides a framework for researchers, journal editors, grant funders, and regulatory authorities to assess the potential clinical relevance of preclinical animal research. We have developed a tool to assess the clinical relevance of preclinical studies. This tool is currently being piloted.


[Genetic basis for skeletal disease. Osteogenesis imperfecta and genetic abnormalities].
Patients with osteogenesis imperfecta (OI) represent various degrees of bone fragility and accompany many clinical manifestations such as dentinogenesis imperfecta, blue sclera, growth disturbance, hearing impairment and so on. Although most OI is caused by genetic mutation of type I collagen gene ; COL1A1 and COL1A2, other genes that concerns post-translational modification of type I collagen molecules such as CRTAP, LEPRE1, PPIB, SERPINH1 and FKBP10 were found to be the causative candidates of OI. On the other hand, genetic causes of type V and type VI OI are not identified. For the classification of OI, Sillence's classification had been used and had been repeatedly revised at the times of identification of new causative genes of OI.


Changes in renal microcirculation in patients with nephrotic and nephritic syndrome: The role of resistive index.
Renal Resistive Index (RRI) is an important and non-invasive parameter of renal damage and it is associated with abnormal microcirculation or to a parenchymal injury. The aim of our study was to compare the RRI in a cohort of patients with renal diseases categorized in three groups: nephrotic syndrome (NS), acute nephritic syndrome (ANS) and patients with urinary abnormalities (UA). Four hundred eighty-two patients with median age of 48 years (IQR 34-62) with indications for kidney disease were included in the study. Biochemical analyses, clinical assessment with detection of NS, ANS and UA and comorbidities were reported. Renal Doppler ultrasound with RRI was evaluated in all patients at the time of enrolment. NS was present in 81 (16.8 %) patients while ANS in 81 (16.8 %) and UA in 228 (47.3 %) patients. Patients with ANS showed significant higher RRI compared to both patients with NS [0.71 (IQR 0.67-0.78) vs 0.68 (0.63-0.73), p < 0.001] and UA [0.71 (0.67-0.78) vs 0.65 (0.61-0.71), p < 0.001]; RRI was higher in NS patients than in patients with UA [0.68 (0.63-0.73) vs 0.65 (0.61-0.71), p < 0.001]. Patients with ANS had significantly lower median estimated glomerular filtration rate (eGFR) compared respectively to NS and UA patients [19.7 ml/min vs 54.8 ml/min and vs 72.3 ml/min, p < 0.001], while renal length was significantly higher in patients with NS compared to both patients with ANS and UA [111.88 mm vs 101.98 mm and vs 106.15, p < 0.001]. Patients with ANS had more frequently hematuria and RRI ≥ 0.70 (p < 0.001) compared to both patients with NS and patients with UA. The multiple regression analysis, weighted for age, showed that RRI inversely correlates with eGFR (β coefficient = -0.430, p < 0.001). Higher and pathological RRI were found in ANS than NS and UA. Renal resistive index in ANS reflects changes in intrarenal perfusion and microvascular dysfunction related to disease characteristics.


Mitochondrial connexin40 regulates mitochondrial calcium uptake in coronary endothelial cells.
Connexins (Cxs) are a group of integral membrane proteins that can form gap junctions between adjacent cells. Recently, it was reported that Cx43 is expressed not only in the plasma membrane but also in the inner mitochondrial membrane and that it regulates mitochondrial functions. Cx40 is predominantly expressed in vascular endothelial cells (ECs) and plays an important role in the electrical propagation between ECs and endothelial/smooth muscle cells. However, it is unknown whether Cx40 is expressed in the mitochondria and what the role of mitochondrial Cx40 is in endothelial functions. We observed in coronary ECs that Cx40 protein was expressed in the mitochondria, as determined by Western blot and immunofluorescence studies. We found that mouse coronary ECs (MCECs) isolated from Cx40 knockout (Cx40 KO) mice exhibited significantly lower resting mitochondrial calcium concentration ([Ca2+]mito) than MCECs from wild-type (WT) mice. After increase in cytosolic Ca2+ concentration ([Ca2+]cyto) with cyclopiazonic acid, calcium uptake into the mitochondria was significantly attenuated in MCECs from Cx40 KO mice compared with WT MCECs. There was no difference in resting [Ca2+]cyto and store-operated calcium entry in MCECs from WT and Cx40 KO mice. We also detected a significant decrease in the concentration of mitochondrial reactive oxygen species (ROS) in Cx40 KO MCECs. Cx40 overexpression in ECs significantly increased resting [Ca2+]mito level and calcium uptake by mitochondria in response to increased [Ca2+]cyto and augmented mitochondrial ROS production. These data suggest that mitochondrial Cx40 contributes to the regulation of mitochondrial calcium homeostasis.


A single daily dose of soybean phytosterols in ground beef decreases serum total cholesterol and LDL cholesterol in young, mildly hypercholesterolemic men.
Consumption of phytosterol-supplemented margarine lowers total plasma cholesterol (TC) and LDL-cholesterol concentrations in older middle-aged hypercholesterolemic individuals. The effects of incorporating phytosterols into lower-fat foods on the plasma lipids of young men at increased risk of developing cardiovascular disease have not been studied. We tested the hypothesis that a single daily dose of soybean phytosterols added to ground beef will lower plasma TC and LDL-cholesterol concentrations in mildly hypercholesterolemic young men. In a triple-blind, 4-wk study, 34 male college students with elevated plasma TC (5.85 +/- 0.70 mmol/L), LDL cholesterol (4.02 +/- 0.60 mmol/L), and TC:HDL cholesterol (5.5 +/- 1.2) were randomly assigned to the control (ground beef alone) or treatment (ground beef with 2.7 g of phytosterols) group. The phytosterol mixture was two-thirds esterified and one-third nonesterified and consisted of beta-sitosterol (48%), campesterol (27%), and stigmasterol (21%). Consumption of phytosterol-supplemented ground beef lowered plasma TC and LDL-cholesterol concentrations and TC:HDL cholesterol from baseline by 9.3%, 14.6%, and 9.1%, respectively (P < 0.001). The LDL particle size did not change, suggesting that the decrease was primarily of particle number. The decreases were similar in subjects with (n = 8) and without (n = 9) a family history of premature cardiovascular disease. No significant changes were found in the control group. Phytosterol-supplemented ground beef effectively lowers plasma TC and LDL cholesterol and has the potential to become a functional food to help reduce the risk of cardiovascular disease.


Genome-wide association study for femoral neck bone geometry.
Poor femoral neck bone geometry at the femur is an important risk factor for hip fracture. We conducted a genome-wide association study (GWAS) of femoral neck bone geometry, examining approximately 379,000 eligible single-nucleotide polymorphisms (SNPs) in 1000 Caucasians. A common genetic variant, rs7430431 in the receptor transporting protein 3 (RTP3) gene, was identified in strong association with the buckling ratio (BR, P = 1.6 x 10(-7)), an index of bone structural instability, and with femoral cortical thickness (CT, P = 1.9 x 10(-6)). The RTP3 gene is located in 3p21.31, a region that we found to be linked with CT (LOD = 2.19, P = 6.0 x 10(-4)) in 3998 individuals from 434 pedigrees. The replication analyses in 1488 independent Caucasians and 2118 Chinese confirmed the association of rs7430431 to BR and CT (combined P = 7.0 x 10(-3) for BR and P = 1.4 x 10(-2) for CT). In addition, 350 hip fracture patients and 350 healthy control individuals were genotyped to assess the association of the RTP3 gene with the risk of hip fracture. Significant association between a nearby common SNP, rs10514713 of the RTP3 gene, and hip fracture (P = 1.0 x 10(-3)) was found. Our observations suggest that RTP3 may be a novel candidate gene for femoral neck bone geometry.


Multiple system atrophy--the nature of the beast.
Multiple system atrophy (MSA) is generally considered a rare disease, but may account for up to 10% of patients with Parkinsonism. The profusion of names for this disease, which may present to general physicians, psychiatrists, cardiologists, autonomic specialists, general neurologists and those with a special interest in Parkinsonism (this author's own perspective) or cerebellar disorders, together with ignorance of its protean manifestations, may account for its underrecognition and misdiagnosis. In this article, the history and nosology of the condition are considered, and provisional diagnostic criteria are advanced. The usefulness (or otherwise) of ancillary investigations is addressed, and the shortcomings of current methods of treatment are stressed. As with idiopathic Parkinson's disease, the ultimate goal of eradicating the disease entails better diagnosis in order to establish the cause, and thence to develop a radical treatment capable of preventing or arresting the disease process.


The importation of the black rhinoceros (Diceros bicornis) from Zimbabwe into Australia.
The paper describes a program to import and breed black rhinoceros ex situ at Western Plains Zoo in Australia. Nine rhinoceros (7 females and 2 males) captured in 1992 in Chete National Park, Zimbabwe, were transported to Australia via Cocos Island. The veterinary treatment of the animals before and during quarantine in Zimbabwe and on Cocos (Keeling) Islands is described. Three animals died; an adult male on Cocos Islands and a juvenile male and an adult female at Western Plains Zoo, Dubbo, New South Wales. The juvenile male died as a result of trauma sustained shortly after arrival and the two adults after developing a severe hepatopathy. The group of 6 females and an additional 4 males imported from the USA in 1994 have adapted well to captivity and to the climate and environment of central west New South Wales.


When family practice residents are parents: effect on in-training examination scores.
This study examines the effect of parenthood on family practice residents' knowledge of pediatrics by evaluating their performance on the pediatric portion of the American Board of Family Practice in-training examination. The 1986 examination results for five Army family practice residencies were obtained. A faculty member at each program noted on the test report whether or not each of 109 residents had children at the time of the exam. Overall, residents with children had slightly higher scores than those without children on the pediatric portion of the in-training examination. The exception was the PGY1 group where residents with children scored markedly lower than those residents without children. However, none of these differences were statistically significant. Although these results do not refute the possibility of special insight possessed by physicians who are also parents, these physicians do not appear to have any particular advantage on written tests over residents without children.