Novel mitochondrial tRNA Leu(CUN) transition and D4Z4 partial deletion in a patient with a facioscapulohumeral phenotype.
Point mutations in mtDNA-encoded tRNA genes frequently cause isolated myopathies but rarely cause the facioscapulohumeral phenotype. We report on a patient affected with chronic progressive weakness of facioscapulohumeral/peroneal muscles whose muscle biopsy showed a mitochondrial myopathy. mtDNA direct sequencing and RFLP analysis revealed a heteroplasmic transition T12313C which disrupts a conserved site in the T Psi C stem of the tRNA(Leu(CUN)) gene and fulfills the accepted criteria of pathogenicity. A partial deletion of the nuclear DNA D4Z4 region with residual repeat sizes of 25 kb was also found in the patient and in her mother. This is the first reported case of mitochondrial myopathy/facioscapulohumeral muscular dystrophy (FSHD) "double trouble".


Legionella pneumophila serogroup 14 isolated from patients with fatal pneumonia.
Two Legionella-like organisms, one isolated from postmortem lung tissue and the other from a bronchial aspirate, were shown by growth, physiologic, and genetic characteristics to belong to the species Legionella pneumophila. Subsequent serologic testing indicated that both strains belonged to a new serogroup, serogroup 14.


Inhibition of development of laser-induced choroidal neovascularization with suppression of infiltration of macrophages in Smad3-null mice.
We evaluated the effects of the loss of Smad3 on the development of experimental argon laser-induced choroidal neovascularization (CNV) in mice. An in vitro angiogenesis model was also used to examine the role of transforming growth factor-β1 (TGFβ1)/Smad3 signaling in vessel-like tube formation by human umbilical vein endothelial cells (HUVECs). CNV was induced in eyes of 8-12-week-old B6.129-background Smad3-deficient (KO) mice (n=47) and wild-type (WT) mice (n=47) by argon laser irradiation. Results showed that the size of the CNV induced was significantly smaller in KO mice as compared with WT mice at day 14 as revealed by high-resolution angiography with fluorescein isothiocyanate-dextran. Immunohistochemistry and real-time reverse transcription-polymerase chain reaction of RNA extracted from laser-irradiated choroidal tissues were conducted on specimens at specific timepoints. Invasion of macrophages (F4/80+), but not neutrophils (myeloperoxidase+), and appearance of myofibroblasts (α-smooth muscle actin+) were suppressed in laser-irradiated KO tissues. mRNA expression of inflammation-related factors, that is, vascular endothelial growth factor (VEGF), macrophage-chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) and TGFβ1 in choroidal tissues was suppressed by the loss of Smad3. We then examined the effects of adding a Smad3 inhibitor, SIS3, or an ALK5 inhibitor, SB431542, on tube formation promoted by TGFβ1 or VEGF in HUVECs cocultured with fibroblast feeder. Further addition of SIS3 or SB431542 augmented vessel-like tube formation by HUVECs in the presence of TGFβ1 or VEGF. In conclusion, lack of Smad3 attenuated the growth of laser-induced CNV with suppression of inflammation by macrophages in mice. Because blocking TGFβ1/Smad3 signal stimulated the activity of angiogenesis of HUVECs in vitro, the reduction of CNV in vivo in KO mice is attributed to a decrease in growth factor levels in the tissue by the loss of Smad3.


Enzymatic and toxigenic ability of opportunistic fungi contaminating intensive care units and operation rooms at Assiut University Hospitals, Egypt.
Total of 110 isolates belonging to 8 fungal species collected from intensive care units (ICUs) and operation rooms (ORs) at Assiut University hospitals were examined for their ability to produce some extracellular enzymes and mycotoxins which are considered as important factors involved in for fungal pathogenicity. The results revealed that 73, 92 and 78 out of the 110 tested isolates produced protease, lipase and urease respectively; meanwhile, 77 of the tested isolates exhibited some hemolytic activities. Chromatographic analysis (TLC) of the crude extract of the fungal isolates tested revealed that 79 isolates of them had the ability to produce at least one of these mycotoxic compounds (aflatoxins B1, B2, G1, gliotoxin, fumigillin, T-2, zearalenone, roridin A & E, verrucarin A & J, trichoveroids, satratoxin H & E). These results demonstrate that the opportunistic fungal species isolated from (ICUs) and (ORs) and tested exhibited some enzymatic and mycotoxic ability which are the most effective virulence factors contributing to fungal pathogenicity indicating that the management of infection control unit at Assiut University hospitals must be aware of not only bacterial but also fungal contamination.


Urinary neutral glycosphingolipid analysis of patients with Fabry's disease; rapid isocratic elution from high-performance liquid chromatography as per-o-benzoyl derivatives.
Neutral glycosphingolipids from urinary sediments of six patients with Fabry's disease and 11 members of the family of one propositus were analyzed using high-performance liquid chromatography. Per-o-benzoyl derivatives of GlcCer, LacCer, GbOse3Cer and GbOse4Cer were clearly resolved by a solvent mixture of hexane/dioxane/isopropanol (75:25:1, v/v) on a normal-phase silica column. Using our isocratic solvent system, the analysis was completed within 15 min. The smallest amount of glycolipid that could be detected by HPLC was 50 pmol and a linear response was shown at 230 nm up to 400 pmol. The calculated peak area of GbOse4Cer was higher than those of GlcCer, LacCer and GbOse3Cer. The molar ratios of GbOse3Cer to monohexosyl ceramide (CMH) in the urinary sediments were: Fabry hemizygotes, 36.33 +/- 25.54 (n = 6); heterozygotes, 0.94 +/- 0.50 (n = 4); and controls, 0.11 +/- 0.06 (n = 5). The molar ratio CDH/CMH was also higher in patients (8.42 +/- 6.23) than in controls (0.77 +/- 0.23). The female H was a rare example of a carrier with typical clinical manifestations. From the urinary sediment analysis, females E, G and J were suspected to be Fabry heterozygotes, although no clinical signs were observed at the time of examination.


Saquinavir potentiates itraconazole's antifungal activity against multidrug-resistant Candida auris in vitro andin vivo.
Candida species are highly opportunistic yeasts that are responsible for serious invasive fungal infections among immunocompromised patients worldwide. Due to the increase in drug resistance and incidence of infections, there is an urgent need to develop new antifungals and to identify co-drugs that can sensitize drug-resistant Candida to antifungals. The objective of this study was to assess the effect of saquinavir on the activity of azole antifungals against C. auris. The in vitro interaction of saquinavir and three azole antifungals (itraconazole, voriconazole, and fluconazole) was evaluated against a panel of C. auris isolates. The itraconazole/saquinavir combination exhibited a synergistic (SYN) relationship against all C. auris isolates tested with the fractional inhibitory concentration index ranging from 0.03 to 0.27. Moreover, a time-kill kinetics assay revealed that saquinavir restored the itraconazole's fungistatic activity against C. auris. Furthermore, saquinavir restored itraconazole's antifungal activity against other clinically important Candida species. The mechanistic investigation indicated that saquinavir significantly inhibited efflux pumps, glucose utilization, and ATP synthesis in Candida. Finally, a murine model of C. auris infection was used to evaluate the efficacy of the itraconazole/saquinavir combination in the presence of ritonavir (as a pharmacokinetic enhancer). The combination significantly reduced the fungal burden in the kidneys by 0.93-log10 colony-forming units (88%) compared to itraconazole alone. This study identified that saquinavir exhibits a potent SYN relationship in combination with itraconazole against Candida species, which warrants further consideration.


Disproportionate epicardial dilation after transmural infarction of the canine left ventricle: acute and chronic differences.
The relation between acute disproportionate infarct dilation and late postinfarct left ventricular remodeling was examined by implanting multiple radiopaque epicardial markers in the left ventricle of eight dogs and determining regional surface deformation after acute and chronic transmural infarction. Transmural injury was produced by combining coronary ligation with distal embolization of a rubber polymer. Dogs were anesthetized and studied before and 1 h, 24 h and 1 week after infarction. Marker positions were recorded by rapid biplane cineradiography, and three-dimensional coordinates were reconstructed by a computer-assisted tracking system. Regional deformation was expressed by a local surface area equal to the sum of multiple (three to four) triangles generated by marker triplets. As early as 1 h after infarction, end-diastolic area in the infarct region increased by 20.3 +/- 3.1%, while that in the remote region increased by only 7.9 +/- 3.5%. Both changes and the difference between them were significant. At 24 h after infarction, both territories continued to undergo dilation, this time to a similar extent (additional +10.3% in the remote region and +10.1% in the infarct region), thus maintaining the significant disproportionate infarct dilation. At 1 week, however, the infarct territory remained dilated with a mean end-diastolic area 31.4 +/- 3.1% above control, while that in the remote region returned to a net mean 8.5 +/- 4.7% increase. Thus, the major extent of disproportionate infarct dilation occurs within 1 h after transmural injury and is accompanied by remote dilation as a compensatory response. The extent of further infarct dilation achieved by 24 h is maintained in the chronic infarct, and compensatory mechanisms enable noninjured myocardium to become less dilated.(ABSTRACT TRUNCATED AT 250 WORDS)


On a nonparametric combination method for dependent permutation tests with applications.
This paper deals with a combination method for dependent permutation tests, which is nonparametric with respect to the underlying unknown dependence structure. The method is based on a simulation or resampling procedure, conditional on the data, which provides a simulated estimate of the permutation distribution of any statistic. Applications to some unusual and quite complex testing problems are shown.


Should infants and adults sleep in the same bed together?
Health professionals should educate families about risk factors for accidental asphyxiation in shared-sleeping arrangements.


A novel Rickettsia, Candidatus Rickettsia takensis, and the first record of Candidatus Rickettsia laoensis in Dermacentor from Northwestern Thailand.
Three hundred and forty-four tick samples were collected from vegetation at Taksin Maharat National Park, Tak province, northwestern Thailand. They were morphologically identified and molecularly confirmed by 16S rRNA and COI genes as Dermacentor laothaiensis (n = 105), D. steini (n = 139), and D. auratus (n = 100). These ticks were examined for the spotted fever group rickettsiae (SFGRs) using PCR and DNA sequencing of six genes; 17-kDa, gltA, 16S rRNA, ompA, ompB, and sca4. Of these ticks, 6.10% (21/344) gave positive results for the presence of SFGRs. Phylogenetic analyses of the SFGRs clearly indicated that a novel genotype assigned as Candidatus Rickettsia takensis was detected in D. laothaiensis (19/105) and at lesser frequency in D. steini (1/139). Furthermore, Candidatus Rickettsia laoensis was also found at a low frequency in D. auratus (1/100), the first record in Thailand. Although, the pathogenicities of these SFGRs remain unknown, our findings suggest potential risks of SFGRs being transmitted via ticks near the border between Thailand and Myanmar, a gateway of daily migrations of local people and visitors both legal and illegal.


Isoagglutinin removal by plasma centrifugation or filtration (single or double): A prospective study in a single center.
ABO-incompatible (ABOi) kidney transplantation, a well-established procedure, has good long-term results provided pretransplant desensitization that includes immunosuppression and apheresis. To compare, within the first pretransplant apheresis session given to 29 ABOi kidney-transplant candidates, the effect on isoagglutinin titers (both IgG and IgM isotypes) of three modalities: centrifugation therapeutic plasmapheresis (cTP; n = 10), filtration TP (fTP; n = 9), and double-filtration plasmapheresis (DFPP; n = 10). The three groups were comparable according to baseline demographics. Treated plasma volumes were similar across the three groups, that is, 4111 ± 403 mL (cTP), 3861 ± 282 mL (fTP), and 3699 ± 820 mL (DFPP): that is, 54 ± 7, 53 ± 7, and 53 ± 10 mL/kg respectively. One session of centrifugation or filtration TP reduced IgG anti-A/anti-B isoagglutinin titer by ~4, whereas one DFPP session reduced it by ~2. One session of cTP reduced IgM anti-A isoagglutinin titer by a little less than 4, whereas fTP and DFPP sessions reduced it by ~3. There were no statistical differences across the three groups regarding isoagglutinin rebound (IgG and IgM). However, isoagglutinin IgG rebound was >4 dilutions for anti-B titers compared with ~2 dilutions for anti-A titers. The median decreases in IgG level were -3.9 g/L (DFPP), -5.9 g/L (cTP), and - 6.06 g/L (fTP) (p = ns). Median fibrinogen depletions were ~ 60% (fTP), 64% (DFPP), and 76% (cTP). Isoagglutinin depletions within the first apheresis session were similar across cTP, fTP, and DFPP: this was numerically lower for DFPP.


Beta blocker effects on plasma homocysteine levels in patients with hypertension.
Recent studies have shown that hyperhomocysteinemia might play a role in the pathogenesis of hypertension. The effects of antihypertensive agents on plasma homocysteine levels have not been tested extensively. We investigated the effects of beta-blocker therapy on homocysteine levels in patients with hypertension. In the study, 120 patients with newly diagnosed hypertension were enrolled. All patients received metoprolol succinate 100 mg/day initially. If blood pressure was above normal on the 15th day of follow-up, the metoprolol dosage was doubled. Before initiation of the antihypertensive medication and after the fourth month of treatment, homocysteine levels were measured. Of the 120 patients enrolled, 39 could not complete the study. Homocysteine levels decreased significantly by the end of the fourth month when compared with basal values (13.5+/-4.5 micromol/l versus 12.4+/-4.9 micromol/l; P = 0.001). There was no relation between homocysteine level and blood pressure control. There was a significant decrease in homocysteine levels in the women treated in this study (P = 0.001); however, this effect was absent in men (P = 0.185). We demonstrate that metoprolol succinate treatment significantly decreases plasma homocysteine levels in patients with hypertension, especially in women.


The intracorrelation of family health insurance and job lock.
This paper tests for the presence of job lock and "health insurance plan lock" stemming from the health shock of a child family member. Using the onset of an acute, unanticipated health shock, I estimate a 7-14 percent decreased likelihood of all family members leaving their current health insurance network and health plan within one year of the emergency. This corresponds to a reduced one-year job mobility rate of approximately 13 percent for the health plan's primary policyholder. Furthermore, the non-portability of health insurance products may contribute to the observed job and health plan lock.


Antinociceptive effect of rat D-serine racemase inhibitors, L-serine-O-sulfate, and L-erythro-3-hydroxyaspartate in an arthritic pain model.
N-methyl-D-aspartic acid receptor (NMDAr) activation requires the presence of D-serine, synthesized from L-serine by a pyridoxal 5'-phosphate-dependent serine racemase (SR). D-serine levels can be lowered by inhibiting the racemization of L-serine. L-serine-O-sulfate (LSOS) and L-erythro-3-hydroxyaspartate (LEHA), among others, have proven to be effective in reducing the D-serine levels in culture cells. It is tempting then to try these compounds in their effectiveness to decrease nociceptive levels in rat arthritic pain. We measured the C-reflex paradigm and wind-up potentiation in the presence of intrathecally injected LSOS (100 μg/10 μL) and LEHA (100 μg/10 μL) in normal and monoarthritic rats. Both compounds decreased the wind-up activity in normal and monoarthritic rats. Accordingly, all the antinociceptive effects were abolished when 300 μg/10 μL of D-serine were injected intrathecally. Since no in vivo results have been presented so far, this constitutes the first evidence that SR inhibitions lower the D-serine levels, thus decreasing the NMDAr activity and the consequent development and maintenance of chronic pain.


Frequency of factor V, prothrombin and methylenetetrahydrofolate reductase gene variants in preeclampsia.
The association between thrombophilic variants (Leiden mutation of the factor V gene, G20210A mutation of the prothrombin gene and C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene) with preeclampsia was investigated in a north-eastern Italian population. Fifty-eight preeclamptic (PE) women and 74 normal pregnancies were evaluated. Genotypes were determined by polymerase chain reaction. The frequency of heterozygous carriers of the factor V Leiden was similar between PE women (5.2%) compared to the control subjects (4.1%; p 0.76). Also the frequencies of G20210A and C677T mutations were similar between PE and control subjects. In this population, we found no difference in the prevalence of genetic risk factors for thrombosis in women with preeclampsia compared with control subjects.


Effects of cysteamine on pro-somatostatin related peptides.
Intracerebroventricular (icv) injection of cysteamine to rats produced a marked depletion of somatostatin-14-like immunoreactivity (LI) in all rat brain regions examined. The somatostatin-28 (SS28)-LI and SS28(1-12)-LI were generally not altered by the cysteamine treatment. Following subcutaneous injection of the drug similar depletions of hypothalamic SS14-LI was observed with no change in SS28-LI nor SS28(1-12)-LI. In vitro cysteamine significantly increased the basal release of SS14-LI and markedly potentiated the evoked release of SS14-LI from hypothalamic slices. At 10 mM cysteamine, enhanced release of SS14-LI from hypothalamic slices was still observed despite a marked depletion of tissue content of SS14-LI.


Photoreceptor Cell Injury Starts in the Initial Stage of Vogt-Koyanagi-Harada Disease.
To demonstrate photoreceptor cell injury in Vogt-Koyanagi-Harada (VKH) disease. The SD-OCT and FAF tests of 16 patients diagnosed with VKH disease that was followed up from the initial stage to the convalescent stage were reviewed. In the initial stage of VKH disease, there was hyperautofluerescence around the macular edema. In the convalescent stage of VKH disease, loss of photoreceptor and injury of RPE have been detected by OCT scan at the same area, which was corresponded with the abnormalities in FAF. The VKH disease causes photoreceptor injury starting in the initial stage of the disease.


Functional human NAIP promoter transcription regulatory elements for the NAIP and PsiNAIP genes.
Neuronal apoptosis inhibitory protein (NAIP) has been shown to inhibit apoptosis in vitro and in vivo with an expression which is regulated in a variety of cells and tissues and may be modulated by a variety of external stimuli. To understand the molecular basis of the transcriptional regulation of the NAIP gene, we have analyzed the 5'-flanking region and transcription of the human NAIP gene. The functional promoter and silencer elements were identified by luciferase reporter constructs in transient transfection experiments using four different human cells. Although the location of the functional elements were shared among the different cells used, the activities for the NAIP promoter varied. Further, cell type-specific protein binding activities were observed by an electrophoretic mobility shift assay (EMSA). EMSA analysis with specific antibodies and DNA sequence analysis identified the POU domain transcription factor Brn-2 as a candidate transcriptional regulator of the NAIP gene. The DNA sequence of the promoter region of the PsiNAIP gene, a copy gene for NAIP, was nearly identical to that of the NAIP gene, indicating a common regulatory mechanism for transcription of the NAIP and PsiNAIP genes. Indeed, the transcript of the PsiNAIP gene was identified. These results provided the first evidence for the functional promoter and candidate transcriptional factor for the NAIP gene and transcription of the PsiNAIP gene.


Stochastic simulation of clonal growth in the tall goldenrod, Solidago altissima.
As clonal plants grow they move through space. The movement patterns that result can be complex and difficult to interpret without the aid of models. We developed a stochastic simulation model of clonal growth in the tall goldenrod, Solidago altissima. Our model was calibrated with field data on the clonal expansion of both seedlings and established clones, and model assumptions were verified by statistical analyses.When simulations were based on empirical distributions with long rhizome lengths, there was greater dispersal, less leaf overlap, and less spatial aggregation than when simulations were based on distributions with comparatively short rhizome lengths. For the field data that we utilized, variation in rhizome lengths had a greater effect than variation for either branching angles or "rhizome initiation points" (see text). We also found that observed patterns of clonal growth in S. altissima did not cause the formation of "fairy rings". However, simulations with an artificial distribution of branching angles demonstrate that "fairy rings" can result solely from a plant's clonal morphology.Stochastic simulation models that incorporated variation in rhizome lengths, branching angles, and rhizome initiation points produced greater dispersal and less leaf overlap than deterministic models. Thus, variation for clonal growth parameters may increase the efficiency of substrate exploration by increasing the area covered and by decreasing the potential for intraclonal competition. We also demonstrated that ramet displacements were slightly, but consistently lower in stochastic simulation models than in random-walk models. This difference was due to the incorporation of details on rhizome bud initiation into stochastic simulation models, but not random-walk models. We discuss the advantages and disadvantages of deterministic, stochastic simulation, and random-walk models of clonal growth.


The diagnostic value of kappa/lambda ratios determined by flow cytometric analysis of biopsy specimens in B-cell lymphoma.
Flow cytometry (FC) is widely utilized in the diagnosis of lymphoma and the light chain ratio (LCR) is especially useful in the diagnosis of B-cell malignancy. In this study we analysed, retrospectively, the predictive value of the LCR in the diagnosis of B-cell lymphoma in 105 consecutive patients with persistent lymph node enlargement or extranodal masses who underwent biopsy. We used a receiver-operating characteristic curve to establish a LCR threshold value of 2.0. The specificity, sensitivity, positive and negative predictive values were 92.3%, 73.1%, 90% and 77%, respectively. We concluded that determination of LCR is a useful adjunct to pathological diagnosis.


Do siblings of myocardial infarction patients have a specific management of hypertension?
The aim of this study was to determine whether the management of hypertension differs between siblings of myocardial infarction patients and the general population. Siblings aged 35 to 74 years, unaffected by myocardial infarction, were drawn from the Augsburg Family Heart Study, conducted in 1996-1997 in southern Germany (n = 524). The reference group consisted of participants of the third MONICA population-based survey conducted in 1994-1995 in the same area, who were aged 35 to 74 years and also unaffected by myocardial infarction (n = 3802). Prevalence, awareness, treatment and control of hypertension (defined by blood pressure > or = 140/90 mm Hg or use of antihypertensive medication) were compared between the two groups. The result was that the prevalence of hypertension was higher in the siblings (men: age-adjusted OR = 1.31, 95% CI: 0.99-1.75; women: age-adjusted OR = 1.83, 95% CI: 1.39-2.41). Male hypertensive siblings were more often aware and treated for hypertension than male hypertensives of the reference group whereas the level of awareness and treatment was comparable between female hypertensives of the two groups. In both genders, no difference in the degree of control was shown between hypertensives of the two groups. In conclusion the siblings and their physicians should pay more attention to the family history of myocardial infarction in order to improve the management of hypertension in this high risk group.


Induction of specific histamine release from basophils with purified natural and recombinant birch pollen allergens.
As much as 15% of the population in industrialized countries suffers from type I allergic symptoms (rhinitis, conjunctivitis, and bronchial asthma). One approach toward this disease involves the production of recombinant allergens in Escherichia coli and their purification for diagnostic and therapeutic purposes. In this study we compared the IgE-binding capacity of natural and recombinant birth allergens with their functional ability to release histamine from allergic patients' basophils via cross-linking of high-affinity Fc epsilon-receptors. The recombinant as well as pollen-derived Bet v I and birch profilin (Bet v II) were purified and tested in parallel on IgE immunoblots and in in vitro histamine release tests (n = 21). We observed an excellent correlation between allergen-induced histamine release and birch pollen RAST (r = 0.881, p < 0.002). Nonspecific histamine release as a result of a cell toxic effect of the allergen preparations was never observed from basophils of donors without birch pollen allergy. The specificity of the presented effector model is also documented by the specific desensitization of patients' basophils with the recombinant allergens. These data may provide a basis for the use of purified recombinant allergens in sensitive and specific in vitro allergy tests monitoring the effector situation in allergic patients, which therefore may represent a possible alternative for in vivo diagnostic methods.


[Microsurgical reconstruction of the burned upper extremity].
Free tissue transplantation is a rarely indicated procedure in burn reconstruction. As the versatility and variability of free flaps have significantly increased during recent years, so too have the indications for these procedures expanded. We retrospectively report the results of 42 free flaps for upper extremity reconstruction in 35 severely burned patients using 13 different free flaps. This experience has enabled us to establish reconstructive principles pertinent to the type of injury (burn versus high voltage injuries) and the timing of reconstruction procedures. In high voltage injuries (n = 17) early free flap coverage (< 21 days after trauma) with muscular flaps was the most frequently used type of reconstruction. Reconstruction site was predominately the forearm. In burn injuries (flame, contact, fluid), free flap coverage was performed during a later stage of the treatment course (3 to 6 weeks after trauma), or as a secondary procedure. Reconstruction with cutaneous or fascial flaps was the preferred method. The elbow and the dorsum of the hand underwent defect coverage in most circumstances. For the reconstruction of complex or large defects (n = 6) combined "chimeric" flaps, preexpansion of free flaps, or the combination of a free and local flap were used. Overall, the flap failure rate was 12 % (n = 5). Interestingly, there was a relationship between flap failure rate and timing of the procedure. Four out of five flap failures occurred within 5 to 21 days after trauma, all five flap failures occurred between five days and six weeks. No flap failure was seen during secondary reconstruction. Our data demonstrate that burn and high voltage injuries are distinct entities, each requiring custom-tailored reconstructive solutions for limb salvage. Even if our flap failures all occurred during the first six weeks it should not be forgotten that this type of coverage is the only alternative to amputation in selective cases.


Ultrastructural variation of Blastocystis hominis stocks in culture.
An ultrastructural study of 10 different Blastocystis hominis stocks was undertaken. Three distinct morphological forms, vacuolar, granular and amoeboid, were distinguished. Numerous variations in the organelles and general cell structure were observed between stocks. B. hominis displayed considerable size variation in the vacuolar forms, ranging from 4 to 63 micron. Thickness and density of the surface coat varied between different stocks. Beneath the surface coat the bilaminar cell membrane displayed electron-dense pits. The nature and quantity of the vacuolar contents varied, and in the granular form four morphologically different inclusions were seen. The organelles which showed the greatest variation between stocks were the mitochondria, varying in shape, electron-density, type of cristae and presence of inclusions. There was minimal variation between stocks with regard to endoplasmic reticulum, Golgi complex and nuclei. Budding of material between the cytoplasm and central vacuole was observed in some stocks. Indications of phagocytic behaviour of B. hominis were seen in the amoeboid form and in the vacuolar form of one stock.


Community management of heart failure.
To review therapies for treating patients with heart failure (HF). Recommendations in this paper are mainly based on the results of randomized controlled trials. To a lesser extent, data from smaller, more physiologic studies are included. Where appropriate, recommendations are based on the results of a consensus conference. Although pharmacologic therapy is the main strategy for treating HF patients, general measures, such as counseling and advice about regular physical activity, are an important component of management. Use of angiotensin-converting enzyme inhibitors (ACE-I) is central to treating HF patients, because these agents decrease mortality and morbidity significantly. Digoxin does not reduce mortality but does reduce morbidity. Angiotensin II antagonists, although found to provide clinical benefit equal to ACE-I, have not been found as yet to have similar effects on mortality and morbidity. Diuretics and nitrates are useful for treating these patients' symptoms. Calcium channel blockers should generally be avoided. Angiotensin-converting enzyme inhibitors are the therapy of choice for HF patients and should be used in all cases unless there are contraindications or clear evidence of intolerance. All other therapies are used mainly for symptom relief.


Surgery clerkship evaluations drive improved professionalism.
To determine whether a brief student survey can differentiate among third-year clerkship student's professionalism experiences and whether sharing specific feedback with surgery faculty and residents can lead to improvements. Medical students completed a survey on professionalism at the conclusion of each third-year clerkship specialty rotation during academic years 2007-2010. Comparisons of survey items in 2007-2008 revealed significantly lower ratings for the surgery clerkship on both Excellence (F = 10.75, p < 0.001) and Altruism/Respect (F = 15.59, p < 0.001) subscales. These data were shared with clerkship directors, prompting the surgery department to discuss student perceptions of professionalism with faculty and residents. Postmeeting ratings of surgery professionalism significantly improved on both Excellence and Altruism/Respect dimensions (p < 0.005 for each). A brief survey can be used to measure student perceptions of professionalism and an intervention as simple as a surgery department openly sharing results and communicating expectations appears to drive positive change in student experiences.


Selective grazing by the mud snail Ilyanassa obsoleta.
Mud snails (Ilyanassa obsoleta) starved for 48 h were allowed to feed on sediments in laboratory microcosms. Sediment cores sliced at 2 mm intervals were compared to snail stomach contents for per cent carbon and nitrogen, plant pigment contents and species composition of benthic diatoms. Concentrations of carbon, nitrogen, phaeopigments, phycocyanin and chlorophyll were enriched in the top 2 mm of the sediments compared to 7-10 mm depth by a factor of 2-10. In turn, these materials were 20-40 times more concentrated in snail guts than in the surface sediments. Snail feces were enriched for carbon and nitrogen by 5-7 times over the surface sediments. Bacterial chlorophyll peaked at about 3-4 mm in the sediments and was not detectable in the snail stomach contents. The C/N ratio of the snail stomach contents was only 6 compared to a ratio of 8.5 for their feces and 12 for the surface sediments.The percentage of migratory diatoms (e.g. Nitzschia and Navicula) decreased with depth where non-migratory species, such as Fragilaria pinnata, dominated. These migratory species were more common in the snails than in the sediments on which they were feeding.A comparison of daily ingestion rates to the animal's energy budget shows that this selective ingestion is sufficient to meet Ilyanassa's energy needs.


Effects of aging on hepatic IGF-I signaling.
Little is known regarding hepatic insulin-like growth factor-1 IGF-I signaling with aging despite the observation that other tissues demonstrate resistance to IGF-I with aging and declines in liver mass accompany aging. Our aim was to determine if the IGF-I-induced signaling process changes with aging. Young (5 months) and old (24 months) C57BL/6 mice hepatic tissues and blood samples were taken 20 min after an intraperitoneal injection of desIGF-I. Age had no significant effect on plasma glucose, insulin and total IGF-I levels. IRS-1 protein was significantly decreased (33%) with aging. Basal phosphorylation of IRS-1, PKB and ERK were unaffected whereas basal phosphorylation of CREB and FKHR were significantly increased (37 and 33%, respectively) with aging. desIGF-I caused a significant decrease in plasma glucose concentrations in both young (53%) and old (44%) mice. desIGF-I administration significantly increased the phosphorylation of IRS-1 in both young (104%) and old (89%) hepatic tissues. Similarly, the phosphorylation of PKB was dramatically enhanced in both young (527%) and old (350%) hepatic tissues after desIGF-I stimulation. By contrast, desIGF-I administration had no significant effects on the phosphorylation of ERK and phosphorylation of transcription factors CREB and FKHR in both young and old hepatic tissues. These data suggest that aging dose not impair IGF-I signaling in hepatic tissues.


The improvement of oxidative stress by two proprietary herbal medicines in type 2 diabetes.
To evaluate the impact to oxidative stress, atherosclerosis and macrovascular disease by two proprietary herbal medicines including Ginkgo Leaf Tablets and Liuwei Dihuang Pills in type 2 diabetes. The recruited 140 type 2 diabetes were randomly divided into the treatment group and control group which were both received basic diabetic management including anti-hyperglycemia, anti-hypertension, life style adjustment and health education etc. Additionally, the treatment group was given both Ginkgo Leaf Tablets and Liuwei Dihuang Pills while the control group was given placebos of Ginkgo Leaf Tablets and Liuwei Dihuang Pills. The relative clinical indexes about macrovascular events occurrence, atherosclerosis degree(IMT levels), oxidative stress in vivo(plasma carboxymethyl lysine(CML) and 8-isoprostane(8-IsoP) levels), plasma glucose, plasma lipid, blood pressure, other drugs usage situations and so on of two groups before and after consecutive 36-month treatment were accurately collected and statistically analyzed. There were no significant differences of cardiovascular disease, cerebrovascular disease, IMT levels, plasma CML and 8-IsoP levels between the two groups before treatment. After 36-month treatment, the plasma CML and 8-IsoP levels of treatment group were both significantly lower than control group (CML: 312.4 ± 90.4 ng/ml versus 463.5 ± 97.2 ng/ml, P < 0.0001; 8-IsoP: 23.7 ± 9.5 pg/ml versus 62.6 ± 16.1 pg/ml, P < 0.0001) although this improvement was not shared with IMT and macrovascular events. Ginkgo Leaf Tablets and Liuwei Dihuang Pills are beneficial to oxidative stress which plays important role in diabetic atherosclerosis and macrovascular complications. The preventive and therapeutic values of herbal medicines will be proved in further diabetic complication researches.


Extracorporeal photopheresis: Lighting the way to immunomodulation.
Photopheresis, initially established as an effective treatment of cutaneous T-cell lymphoma, has in recent years also been used to treat chronic graft vs. host disease, heart transplant rejection, and several other conditions requiring immunosuppression. Despite reported beneficial results of this procedure in treatment of various conditions, randomized controlled clinical trials are lacking for the majority of suggested indications. Furthermore, the mechanisms of action of this procedure are still unclear. Deeper understanding of the molecular basis of photopheresis-based immunomodulation will allow better selection of patients to be treated and will facilitate development of novel, minimally toxic immunomodulatory treatments.


What makes narratives feel right? The role of metacognitive experiences.
Conviction Narrative Theory holds that reasoners adopt "a narrative that feels 'right' to explain the available data" and use "that narrative to imagine plausible futures" (target article, Abstract). Drawing on feelings-as-information theory, this commentary reviews the role of metacognitive experiences of ease or difficulty and highlights that fluently processed narratives are more likely to "feel right."


Estimating Breast Cancer Overdiagnosis After Screening Mammography Among Older Women in the United States.
Overdiagnosis is increasingly recognized as a harm of breast cancer screening, particularly for older women. To estimate overdiagnosis associated with breast cancer screening among older women by age. Retrospective cohort study comparing the cumulative incidence of breast cancer among older women who continued screening in the next interval with those who did not. Analyses used competing risk models, stratified by age. Fee-for-service Medicare claims, linked to the SEER (Surveillance, Epidemiology, and End Results) program. Women 70 years and older who had been recently screened. Breast cancer diagnoses and breast cancer death for up to 15 years of follow-up. This study included 54 635 women. Among women aged 70 to 74 years, the adjusted cumulative incidence of breast cancer was 6.1 cases (95% CI, 5.7 to 6.4) per 100 screened women versus 4.2 cases (CI, 3.5 to 5.0) per 100 unscreened women. An estimated 31% of breast cancer among screened women were potentially overdiagnosed. For women aged 75 to 84 years, cumulative incidence was 4.9 (CI, 4.6 to 5.2) per 100 screened women versus 2.6 (CI, 2.2 to 3.0) per 100 unscreened women, with 47% of cases potentially overdiagnosed. For women aged 85 and older, the cumulative incidence was 2.8 (CI, 2.3 to 3.4) among screened women versus 1.3 (CI, 0.9 to 1.9) among those not, with up to 54% overdiagnosis. We did not see statistically significant reductions in breast cancer-specific death associated with screening. This study was designed to estimate overdiagnosis, limiting our ability to draw conclusions on all benefits and harms of screening. Unmeasured differences in risk for breast cancer and differential competing mortality between screened and unscreened women may confound results. Results were sensitive to model specifications and definition of a screening mammogram. Continued breast cancer screening was associated with greater incidence of breast cancer, suggesting overdiagnosis may be common among older women who are diagnosed with breast cancer after screening. Whether harms of overdiagnosis are balanced by benefits and for whom remains an important question. National Cancer Institute.


A crosslinked β-cyclodextrin polymer used for rapid removal of a broad-spectrum of organic micropollutants from water.
Organic micropollutants in aquatic environment such as plasticizer, pesticide and pharmaceuticals, have posed a serious threat to human health and are emerging as a great challenge to humanity. Traditional water treatment techniques fail to achieve high removal efficiency for low concentration of organic micropollutants. Here we demonstrate a water-insoluble crosslinked β-cyclodextrin (β-CD) polymer able to remove a broad-spectrum of organic micropollutants from water by rapid adsorption. A family of β-CD polymers (β-CDPs) were synthesized by nucleophilic aromatic substitution of β-CD hydroxyl groups and 4,4'-difluorodiphenylsulfone. The β-CDPs were used to adsorb various organic micropollutants in water by static or dynamic adsorption process. It was found that more than 99% micropollutants in water were removed by flowing the feed water through the column of β-CDPs. The results of static adsorption experiments indicated the adsorption process was fast and the adsorption capacity was very high (the maximal value was 113.0mg of bisphenol A per gram of β-CDP). The adsorption process was fitted well with the quasi-second-order kinetics and the Langmuir isothermal adsorption model, suggesting that it is mainly a chemical adsorption of monolayer. The water-insoluble characteristic of the β-CDPs is convenient for their separation from the treated solution after adsorption saturation for regeneration and reuse. The adsorption ability of β-CDPs was kept nearly unchanged after five filtration-regeneration cycles.


Transatlantic Multispecialty Consensus on Fundamental Endovascular Skills: Results of a Delphi Consensus Study.
The aim of this study was to establish a consensus on Fundamental Endovascular Skills (FES) for educational purposes and development of training curricula for endovascular procedures. The term "Fundamental Endovascular Skills" is widely used; however, the current literature does not explicitly describe what skills are included in this concept. Endovascular interventions are performed by several specialties that may have opposing perspectives on these skills. A two round Delphi questionnaire approach was used. Experts from interventional cardiology, interventional radiology, and vascular surgery from the United States and Europe were invited to participate. An electronic questionnaire was generated by endovascular therapists with an appropriate educational background but who would not participate in subsequent rounds. The questionnaire consisted of 50 statements describing knowledge, technical, and behavioral skills during endovascular procedures. Experts received the questionnaires by email. They were asked to rate the importance of each skill on a Likert scale from 1 to 5. A statement was considered fundamental when more than 90% of the experts rated it 4 or 5 out of 5. Twenty-three of 53 experts invited agreed to participate: six interventional radiologists (2 USA, 4 Europe), 10 vascular surgeons (4 USA, 6 Europe), and seven interventional cardiologists (4 USA, 3 Europe). There was a 100% response rate in the first round and 87% in the second round. Results showed excellent consensus among responders (Cronbach's alpha = .95 first round; .93 second round). Ninety percent of all proposed skills were considered fundamental. The most critical skills were determined. A transatlantic multispecialty consensus was achieved about the content of "FES" among interventional radiologists, interventional cardiologists, and vascular surgeons from Europe and the United States. These results can serve as directive principles for developing endovascular training curricula.


Intravoxel incoherent motion diffusion-weighted MRI for characterizing regional variability and monitoring serial changes of parameters in rabbit VX2 liver tumors.
To characterize intravoxel incoherent motion (IVIM) parameters in different regions of interest (ROIs) of rabbit VX2 liver tumors during a follow-up of 4 weeks. This study was approved by the Institutional Animal Care and Use Committee. In 18 VX2 liver tumor-bearing rabbits, IVIM DW imaging was serially performed at the 2nd, 3rd, and 4th week separately after tumor implantation at 1.5T by using 12 b values. Three ROIs were drawn on the apparent diffusion coefficient (ADC) maps, including the whole lesion, the visibly most restricted diffusion area (MRDA), and peripheral area of tumor. For each ROI of tumors, serial changes in ADC and D, D*, and f at three timepoints were calculated. The differences of IVIM parameters in different ROIs of tumors at each timepoint were compared. Serial measurements of f in all three ROIs (P = 0.000, P = 0.002, and P = 0.000), ADC for the whole lesion, MRDA (P = 0.000 and P = 0.024), and D for MRDA (P = 0.001) from the 2nd to 4th week illustrated a statistical difference. The overall comparison of ADC (P = 0.000, P = 0.014, and P = 0.000), D (P = 0.000, P = 0.001, and P = 0.000), and f (P = 0.001, P = 0.001, and P = 0.000) for three tumor regions at weeks 2, 3, and 4 showed significant differences. D* in all three ROIs at each timepoint showed no significant differences (P = 0.612, P = 0.723, and P = 0.699; 95% confidence interval: 0.184-0.950). The peripheral area of tumor could be used as an alternative to the whole lesion as ROIs to assess the VX2 liver tumor with IVIM DW imaging, and in the period between weeks 2 and 3.


Multi-institutional external validation of seminal vesicle invasion nomograms: head-to-head comparison of Gallina nomogram versus 2007 Partin tables.
The Partin tables represent one of the most widely used prostate cancer staging tools for seminal vesicle invasion (SVI) prediction. Recently, Gallina et al. reported a novel staging tool for the prediction of SVI that further incorporated the use of the percentage of positive biopsy cores. We performed an external validation of the Gallina et al. nomogram and the 2007 Partin tables in a large, multi-institutional North American cohort of men treated with robotic-assisted radical prostatectomy. Clinical and pathologic data were prospectively gathered from 2,606 patients treated with robotic-assisted radical prostatectomy at one of four North American robotic referral centers between 2002 and 2007. Discrimination was quantified with the area under the receiver operating characteristics curve. The calibration compared the predicted and observed SVI rates throughout the entire range of predictions. At robotic-assisted radical prostatectomy, SVI was recorded in 4.2% of patients. The discriminant properties of the Gallina et al. nomogram resulted in 81% accuracy compared with 78% for the 2007 Partin tables. The Gallina et al. nomogram overestimated the true rate of SVI. Conversely, the Partin tables underestimated the true rate of SVI. The Gallina et al. nomogram offers greater accuracy (81%) than the 2007 Partin tables (78%). However, both tools are associated with calibration limitations that need to be acknowledged and considered before their implementation into clinical practice.


[Arthroderma benhamiae strains in Germany : Morphological and physiological characteristics of the anamorphs].
Anamorphs of Arthroderma (A.) benhamiae, which can cause inflammatory tinea lesions in humans, have been progressively spreading in Germany. Identification of A. benhamiae anamorphs by conventional methods. Evaluation of our own results obtained with A. benhamiae anamorphs and from the relevant literature. Infections with A. benhamiae anamorphs are usually transferred by guinea pigs or other animals. A. benhamiae anamorphs can be cultured on growth media used for dermatophytes and can be characterized morphologically and physiologically. In Germany the yellow variant is seen most often but a white variant that equals Trichophyton (T.) interdigitale has also been observed. On Sabouraud agar the yellow variant develops markedly yellow thalli, whereas the white variant produces white aerial mycelium. Microconidia are formed by the yellow variant-if at all-only scarcely and delayed; they are small, roundish, and arranged in a grape-like order. The white variant produces peg-shaped microconidia alongside hyphae as well as roundish ones in grape-like clusters, and subsequently some macroconidia, chlamydospores, and spiral hyphae. In microcultures with both variants circuit-like hyphal structures can consistently be demonstrated. On Trichophyton agars only the yellow variant is clearly dependent on thiamine. The urease test is negative with the yellow variant and positive with the white variant. Most strains of both variants are negative in the hair perforation test. The characteristics described allow reliable identification of the yellow variant of the A. benhamiae anamorph by conventional methods; a distinction between the white variant and T. interdigitale can be more difficult. Dermatologists should be able to identify this agent in clinical routine.


Whole-Genome Sequencing Analysis of Chromobacterium piscinae Strain ND17, a Quorum-Sensing Bacterium.
Here, we report the draft genome sequence of Chromobacterium piscinae strain ND17. This bacterium was isolated from a fresh water sample in Malaysia and exhibits quorum-sensing activity. This first draft genome of C. piscinae strain ND17 will pave the way to future studies of the quorum-sensing properties of this isolate.


Renovascular reconstruction: factors affecting long-term prognosis in 919 patients followed up to 31 years.
During the 31-year period from May 3, 1955, to May 12, 1986, renovascular reconstructions were performed on 919 patients. The mean age of the 529 men was 54 +/- 0.58 (SE) years and, of the 390 women, 48 +/- 0.7 years. Mean preoperative diastolic blood pressure was 110 +/- 0.6 mm Hg. The most common causes of renal artery stenosis were atherosclerosis in 647 patients, fibromuscular disease in 161 patients, and renal artery aneurysm in 51 patients. In the remaining 60 patients, other causes were present, including kinks and fibrous bands. The most common surgical procedures were Dacron bypass graft (780 arteries) and endarterectomy with or without a patch graft (329 arteries). Four hundred sixty-nine patients had associated operations, the most common of which were abdominal aortic aneurysmectomy in 231 and aortoiliofemoral reconstruction in 141 patients. The perioperative mortality rate was 5.5% (51 of 919 overall); for renal procedures alone, it was 1.7% (8 of 450) and for combined surgical procedures, 9.2% (43 of 469). The overall graft patency rate at a follow-up of 18.8 +/- 1.9 months was 88.6% (381 of 430) and at a second follow-up of 50 +/- 4.3 months, 86.7% (111 of 128). Analysis of long-term blood pressure response and factors affecting late survival indicated that patients with preoperative diastolic pressures of greater than 100 mm Hg and renal artery stenosis of greater than 70% had the best blood pressure responses and that male sex, increasing age, bilateral renal stenosis, and associated vascular operations lowered the survival rate whereas fibromuscular disease enhanced the duration of survival.(ABSTRACT TRUNCATED AT 250 WORDS)


Penetrating high-velocity genitourinary injuries. Part II: Ureteral, lower tract, and genital wounds.
Penetrating ureteral wounds appear to occur at a constant low rate and demand internal splinting without ureterotomy drainage in the multiple-injured patient. Concomitant upper urinary tract and colonic or pancreatic injury absolutely requires the use of nephrostomy drainage. Most prostatic or prostatomembranous rupture can be approached retropubically, or if extensive tissue damage and uncontrollable hemorrhage are present, by transpubic methods. Intravesical fixation of the prostatomembranous rupture provides the best anchorage and approximation. Rectoprostatic-bladder wounds should be approached suprapubically and may be drained transcoccygeally also. Section of the obturator nerve may provide postoperative pain relief in severe pubic ramus fractures. Small high-velocity entrance wounds in the external genitalia warrant exploration because of great internal disruption. The skin of the external genitalia should be closed primarily.


North American single-center experience with a sutureless aortic bioprosthesis.
Surgical sutureless aortic valves have the potential for shorter procedural times and could benefit patients with increased risk. The Enable (Medtronic Inc, Minneapolis, Minn) valve is a bioprosthesis housed in a Nitinol cage allowing folding and deployment once implanted. We aimed to evaluate the early clinical and echocardiographic results with the Enable valve. Patients with aortic stenosis, Society of Thoracic Surgeons score greater than 5.0%, the need for combined procedures, and frailty were considered for Enable implantation. Between August 2012 and October 2014, 63 patients underwent implantation of the Enable valve (Medtronic Inc, Minneapolis, Minn). Thirty patients underwent isolated aortic valve replacement. Combined procedures were aortic valve replacement/coronary artery bypass grafting (26), aortic valve replacement/mitral valve repair (2), aortic valve replacement/mitral valve repair/coronary artery bypass grafting (2), and aortic valve replacement/ascending aortic graft (3). Predicted Society of Thoracic Surgeons score was 8.06 ± 7.73 (0.94-41.30). Implant success was 100%. Mean crossclamp time for isolated aortic valve replacement was 44 ± 14 minutes (30-91). Thirty-day mortality was 1.6% (1/63), and late mortality was 3.2% (2/62). No mortality was valve related. Intraoperative need for revision was 6.3% (4/63). Early migration requiring reoperation occurred in 1.6% of patients (1/63). Postoperative indexed aortic valve area was 1.08 ± 0.22 cm(2)/m(2), and peak and mean gradients were 17 ± 7 mm Hg and 9 ± 4 mm Hg, respectively. The rate of complications was as follows: pacemaker 3.1% (2/63), transient ischemic attack 1.6% (1/63), other thromboembolic events 0%, bleeding 0%, and endocarditis 0%. Mean follow-up was 10 ± 8 months. At latest follow-up, 61 patients were in New York Heart Association class I. Moderate or severe aortic regurgitation did not develop in any patients in the follow-up period. The Enable bioprosthesis is an acceptable alternative to conventional aortic valve replacement in higher-risk patients. The early hemodynamic performance seems favorable.


K-score: A novel scoring system to quantify fatigue-related ergonomic risk based on joint angle measurements via wearable inertial measurement units.
Work-related musculoskeletal disorders have been recognized as a global problem that affects millions of people annually. Fatigue is one of the main contributors to musculoskeletal disorders. Thus, this study investigated fatigue detection based on the measured body motion by wearable inertial measurement units. We quantified the body motion during manual handling tasks using a novel kinematic score (i.e., K-score), and the Rapid Entire Body Assessment (REBA). K-score and REBA were calculated using joint angles. Nevertheless, unlike REBA, K-score showed a significant correlation (Spearman's correlation coefficient of ρ(302) = 0.21, p < 0.05) with electromyography (EMG) signal amplitude, which was affected by muscle fatigue. Therefore, in-field measurement of K-score using inertial measurement units could detect the fatigue-induced change of body motion in long-duration manual handling tasks. Our proposed K-score can be used to assess fatigue-related ergonomic risk in long-term and real-world working conditions without the need for tedious EMG recording at workplaces.


3-D reconstruction of bluetongue virus tubules using cryoelectron microscopy.
Bluetongue virus (BTV) forms tubules in infected mammalian cells. These tubules are virally encoded entities which can be formed with only one protein, NS1. The NS1 protein does not form a part of virus particles, and its function in viral infection is uncertain. Expression of the NS1 gene in insect cells by recombinant baculovirus yields high amounts of NS1 tubules (ca. 50% of cellular proteins) which are morphologically and immunologically similar to authentic BTV NS1 and can be isolated to about 90% purity. The structure of these synthetic NS1 tubules was investigated by cryoelectron microscopy. NS1 tubules are on average 52.3 nm in diameter and up to 100 nm long. The structure of their helical surface lattice has been determined using computer image processing to a resolution of 40 A. The NS1 protein is about 5.3 nm in diameter and forms a dimer-like structure, so that the tubules are composed of helically coiled ribbons of NS1 "dimers," with 21 or 22 dimers per turn. The surface lattice displays P2 symmetry and forms a one-start helix with a pitch of 9.1 nm. The NS1 tubules exist in two slightly different pH-dependent conformational states.


Patients' noncompliance and how doctors manage this.
The purpose of this study was to examine doctors' behavior towards patients' noncompliance and to explain it within the framework of frustration theory. Two groups of physicians participated. The first group of 30 was selected from a larger group, based on tape-recordings of patients' visits where obvious examples of noncompliance were found. The 63 physicians forming the second group were interviewed to discover their attitude/behavior towards noncompliance. The tactics observed in the first study group and reported by the second group were then analysed according to the same system of categories. In the second group RPFS (Rosenzweig Picture-Frustration Study) was also applied to find determinants of the doctors' behavior. In general, 11 different tactics were identified but only five were applied by more than 10% of doctors from either group. These were: medical threat (most frequently used in both groups) carrying the doctor's point in an indulgent atmosphere, authoritarian tactics, medical information, withdrawal. There were no significant differences between the groups as to the tactics used (i.e. no differences between observed and reported behavior), on the contrary, a positive correlation was found. The doctors applied certain tactics of doubtful effectiveness which might worsen the patient's emotional state or harm him as a person. On the other hand, many valuable techniques of proved efficacy were not used. When the doctors' tactics were interpreted within the framework of frustration theory as task-oriented or ego-defensive, it was found that in both groups the ego-defensive tactics predominated. The kind of tactics adopted towards noncompliance was strongly determined by the attitude towards frustration measured by RPFS (Kendall Q coefficient = 0.92).(ABSTRACT TRUNCATED AT 250 WORDS)


The threshold of insulin-induced hypophagia is lower in chicks selected for low rather than high juvenile body weight.
Chicks genetically selected for low juvenile body weight had a lower threshold of central insulin-induced decreased food and water intake and whole blood glucose concentration than those selected for juvenile high body weight. Plasma corticosterone concentration was increased but not differently between lines. Therefore, selection may have affected insulin sensitivity which may have then contributed to their hypo- and hyperphagia and differential body weights.


Reducing the burdens of forced abortion travel: Referrals, financial and emotional support, and opportunities for positive experiences in traveling for third-trimester abortion care.
In the United States, travel is a fact of the abortion care provision landscape. This is largely due to the uneven geographical distribution of providers and state-level gestational duration bans that constrain what abortion care is available locally. When abortion travel is compelled by legal restriction, it is forced travel. Research has comprehensively documented that forced abortion travel is burdensome; people who must travel for abortion experience financial, logistical, and emotional burdens. Generally overlooked, however, is variation in the experience of travel-related burdens and whether and how such burdens can be reduced. Given current political hostility to abortion, the number of people who must travel and the distances they must travel for abortion are likely to grow, making the question of how travel-related burdens can be reduced in the absence of policy change of increasing relevance. Using thematic analysis of semi-structured interviews with 30 cisgender women in the United States who were forced to travel to obtain third-trimester abortion care, I identify three ways that the burdens of forced abortion travel can be mitigated without policy change: prompt referrals; financial and practical support for travel; and emotional support. In some instances, respondents experienced the received emotional support as so valuable as to offset the other burdens of travel, pointing to the possibility that some people might prefer to travel for abortion care whether or not they are forced to do so. Respondents also reported unexpected positive aspects of traveling, including experiences of kindness and human connection, underscoring that not all aspects of abortion travel are negative. Findings thicken our understanding of forced abortion travel and identify structural and interpersonal practices that can reduce the associated burdens, complementing legal and policy-oriented critiques of legal regulation that makes abortion travel necessary.


"Unraveling the tail" of how SRPK1 phosphorylates ASF/SF2.
In this issue of Molecular Cell, Ngo et al. (2008) describe the crystal structure of the SRPK1 protein kinase in complex with its substrate, the spliceosome factor ASF/SF2, providing an unprecedented view of multiple targeting mechanisms in action on a single substrate.


Cardiovascular diseases as target for imaging.
Recent pathophysiological findings have lead to new concepts to identify patients at risk for cardiovascular disease using systemic serum markers or new imaging methodology. New probe technology and progress in imaging techniques have set the base for development of molecular imaging concepts in the cardiovascular systems. The aim of these new imaging techniques is the detection of active biological processes in cardiovascular systems combining specific probes with contrast agents for MRI, SPECT or PET. There are promising strategies mostly in preclinical tests, which will prove clinical applicability in the near future.


Parasites of pufferfish, Lagocephalus spp. and Torquigener flavimaculosus of the Israeli Mediterranean: A new case of Lessepsian endoparasites.
With the opening of the Suez Canal as a link between the Red Sea and the Mediterranean Sea in 1869, the biogeographical event of the Lessepsian migration has been starting. Aided by beneficial conditions in the new habitat, almost 500 marine species have immigrated and often established themselves in the Mediterranean Sea, including several pufferfish species, with all of them extending their range and becoming important components of the local fauna. The parasitic fauna of these pufferfish has scarcely been examined in the Mediterranean Sea or in their native range, which provides the opportunity to study host-parasite interaction in a new habitat. The present study describes the parasitic fauna in four alien invasive pufferfish species (Lagocephalus guentheri, L. sceleratus, L. suezensis, and Torquigener flavimaculosus) of various sizes and ages on the Israeli Mediterranean coast. The parasite fauna of these species was diverse (Maculifer dayawanensis Digenea; Calliterarhynchus gracilis, Nybelinia africana and Tetraphyllidea larvae Cestoda; Hysterothylacium reliquens, Hysterothylacium sp. and Raphidascaris sp. Nematoda; Trachellobdella lubrica Hirudinea and Caligus fugu and Taeniacanthus lagocephali Copepoda) and consisted of mostly generalist species, most likely acquired in the new habitat, and specialist copepod ectoparasites, having co-invaded with the pufferfish. Additionally, the oioxenic opecoelid digenean Maculifer dayawanensis was found in two pufferfish species. The genus was previously only known from the Indo-Pacific Ocean, representing the eighth reported case of a Lessepsian endoparasite so far. Our results suggest a change in parasite fauna to native Mediterranean species in the pufferfish like previously reported in other Lessepsian migrant predatory fish species and a wider spread of co-invasion of fish endoparasites to the Mediterranean Sea than previously assumed. The study also provides several new host records and the first report for parasites in T. flavimaculosus.


The diffusion MRI signature index is highly correlated with immunohistochemical status and molecular subtype of invasive breast carcinoma.
To evaluate the relationship of the signature index (S-index), an advanced diffusion MRI marker, and the immunohistochemical (IHC) status, proliferation rate, and molecular subtype of invasive breast cancers. A retrospective study of patients with invasive carcinoma was conducted between 2017 and 2021. All patients underwent dynamic contrast-enhanced MRI and DWI using a 3-T system. For DWI, three b values (0, 200, and 1500 s/mm2) were used to derive the S-index. Three-dimensional ROIs were manually placed over the whole tumor on DWI. Mean and 85th percentile S-index values were compared to the IHC status, proliferation rate, and molecular subtypes of lesions. The study included 153 patients (mean age, 60 ± 13 years) with 160 invasive carcinomas. S-index values were significantly higher in estrogen receptor-positive (mean, p = .005; 85th percentile, p < .001) and progesterone receptor-positive (mean, p = .003; 85th percentile, p < .001) tumors, and significantly lower in human epidermal growth factor receptor 2 (HER2) - positive tumors (mean, p = .023; 85th percentile, p < .001). Mean and 85th percentile S-index values were significantly different among breast cancer subtypes (mean, p = .015; 85th percentile, p = .002), and the AUC of these values for the prediction of IHC status were 0.64 and 0.66 for HER2, and 0.70 and 0.74 for hormone receptors, respectively. The DWI S-index showed significantly higher values in invasive carcinomas with immunohistochemical status associated with good prognosis, suggesting its usefulness as a noninvasive imaging biomarker to estimate IHC status and orient treatment. • The signature index, an advanced diffusion MRI marker, showed good discrimination of immunohistochemical status in invasive breast carcinomas. • The signature index has the potential to differentiate noninvasively invasive breast carcinoma subtypes and appears as an imaging biomarker of prognostic factors and molecular phenotypes.


A potential human hepatocellular carcinoma inhibitor from Bauhinia purpurea L. seeds: from purification to mechanism exploration.
A 20-kDa Kunitz-type trypsin-chymotrypsin inhibitor, Bauhinia purpurea trypsin inhibitor (BPLTI), has been isolated from the seeds of B. purpurea L. by using liquid chromatography procedures that involved ion exchange chromatography on Sp-Sepharose and Mono S and gel filtration on Superdex 75. BPLTI demonstrated protease inhibitory activities of 7226 BAEE units/mg and 65 BTEE units/mg toward trypsin and α-chymotrypsin, respectively. BPLTI was relatively thermal (0-60°C) and pH (3-10) stable and its activity could be decreased by dithiothreitol treatment. BPLTI exhibited a wide spectrum of anti-proliferative and pro-apoptotic activities especially on human hepatocellular carcinoma Hep G2 cells. However, it was devoid of a significant antiproliferative effect on immortal human hepatic WRL 68 cells. We show here that BPLTI stimulates apoptosis in Hep G2 cells, including (1) evoking DNA damage including the production of chromatin condensation and apoptotic bodies; (2) induction of cell apoptosis/necrosis; (3) mitochondrial membrane depolarization; and (4) increasing the production of cytokines. Taken together, our findings show for the first time that purified protease inhibitor from B. purpurea L. seeds is a promising candidate for the treatment of human hepatocellular carcinoma.


Early and late cardiac ventricular reverse remodeling after catheter ablation for lone paroxysmal atrial fibrillation.
We sought to explore ventricular function in patients with lone paroxysmal atrial fibrillation (AF) and determine the mid- and long-term impact of pulmonary vein isolation on cardiac remodeling. The relationship between tachyarrhythmia and ventricular dysfunction is still a matter of debate. Tachycardia-induced cardiomyopathy is defined as reversible myocardial dysfunction following treatment for tachyarrhythmia. We prospectively studied 31 patients (56.4 ± 10 years) presenting with paroxysmal-AF who were treated successfully by catheter ablation and 15 age-matched controls. Left and right ventricular functions were assessed by echocardiography at baseline and at 3-month and 1-year follow-up. In AF-patients, LV-function was slightly lower at baseline than controls (LV-ejection fraction was 60% versus 64%; P = 0.06). More impressive, systolic peak velocity on Doppler tissue imaging was 9 cm/s in AF patients (versus 12 cm/s; P = 0.0004). LV global longitudinal strain was also significantly different between the two groups (patients: -16% versus controls: -19%; P = 0.005). At 1-year follow-up, most functional parameters significantly improved in the AF-patients and no longer differed from the controls. Right ventricular (RV) function was also depressed in AF patients at baseline. At 1-year follow-up, tissue Doppler showed improvement in RV-S' (+27%, P = 0.007) and RV peak systolic strain (+36%, P<0.0001) and became comparable to controls. We demonstrate that some degree of arrhythmic cardiomyopathy exists in patients presenting with lone paroxysmal-AF. Catheter ablation improved RV and LV functions. Longitudinal function is the most sensitive component of ventricular systole to monitor when looking for this cardiac reverse remodeling.


May-Thurner syndrome will be completed?
The May-Thurner syndrome is a deep venous thrombosis of the left iliofemoral vein due to a compression by the right primitive iliac artery. Some authors suggested that this syndrome could be associated with genetics factors predisposing to thrombophilia. The aim of this article is to assess if the May-Thurner syndrome could be associated with inherited thombophilia particularly the factor V mutation. We exposed three cases of May-Thurner syndrome, all were positive for the presence of factor V Leiden. Some olds syndromes can be revisited and explained by molecular research. In this case, the May-Thurner syndrome could be associated with inherited thrombophilia. This finding could explain the pathology but also open new treatments or guidelines for the management this kind of pathology.


Self-expanding metallic stent improves histopathologic edema compared with transanal drainage tube for malignant colorectal obstruction.
To compare the usefulness of the self-expanding metallic stent (SEMS) with that of the transanal drainage tube (TDT) and emergency surgery after failure of decompression (ESFD) in patients with malignant colonic obstruction (MCO), and to evaluate post-decompression histopathologic changes. From January 2010 to June 2015, 39 patients with MCO received SEMS, TDT, and ESFD. We evaluated the outcomes including success rates of placement, clinical outcomes after decompression, and histopathologic findings of the resected specimens. Technical success rates were 100% for SEMS and 78.9% for TDT. Clinical success rates were 100% for SEMS and 80.0% for TDT. Postoperative ileus was significantly less frequent after SEMS than after TDT (p = 0.014). Histopathologic edema grade was significantly lower for SEMS than for TDT and ESFD (p < 0.0001). There was no significant difference between edema grade and duration of decompression in the TDT group (p = 0.629), while all patients with SEMS were classified in a low edema grade (grade 0-2). The rate of stoma creation was significantly higher in patients with a high edema grade (grade 3) than in those with a low edema grade (grade 0-2) (p = 0.003). There was no microscopic perforation in any group. Significantly greater resolution of histopathologic edema was achieved after placement of SEMS than after placement of TDT. These findings provide an indication of favorable clinical outcomes of SEMS in comparison with TDT and ESFD. This article is protected by copyright. All rights reserved.


Effects of laboratory versus field exercise on leukocyte subsets and cell adhesion molecule expression in children.
In adults, exercise is a powerful and natural stimulator of immune cells and adhesion molecules. Far less is known about these exercise responses during childhood and whether or not exercise in real-life activities of healthy children might influence immune responses. We compared laboratory exercise (10 x 2 min periods of heavy, constant intensity, cycle ergcometer exercise with 1 min rests between exercise in nine subjects, aged 9-15 years) with field exercise (90 min soccer practice in nine different subjects, aged 9-11 years). Blood was sampled before both protocols, 5 min after the 30 min laboratory protocol, and 10-15 min after the 90 min field protocol. Both field and laboratory exercise protocols led to significant (P<0.05) increases in granulocytes, monocytes, and all lymphocyte subpopulations. The mean (SEM) increases were similar for the two protocols except for the significantly greater increase in laboratory compared with field protocols for natural killer cells [142 (39)% vs 12 (16)%, P<0.001] and monocytes [64 (22)% vs 32 (19)%, P<0.001] Both protocols significantly influenced adhesion molecules (such as CD54) which have not been previously studied in children. However, the adhesion molecule CD8+ CD62L increased to a significantly (P < 0.001) greater extent in the laboratory [101 (25)%] versus field [34 (25)%] protocol. Finally, the density of CD632L on lymphocytes significantly decreased with laboratory exercise but showed no change in the field protocol [-20 (3)% vs -3 (3)%, P<0.001]. The rapid and substantial immune response in both laboratory and field protocols suggests that exercise stimulation of the immune system occurs commonly in the real lives of children and may play a role in their overall immune status.


Precocious puberty in adolescent girls: a biomarker of later psychosocial adjustment problems.
We have investigated the hypothesis that stress condition will accelerate pubertal maturation in adolescent girls and that faster maturing girls will display more behavioral problems than their on-time or late-maturing peers during pubertal development. Longitudinal data were collected yearly from 1986 to 1997. Parents of 1039 French-speaking Canadian girls completed a questionnaire about their characteristics as well as those of their family. To predict onset of puberty by time and cumulative risk factors, a Hierarchical Regression Analysis and a Survival Regression Analysis were conducted. Results revealed that: (1) anxiety at 8 years old, lower level of anxiety a year earlier and more adverse family condition predict a younger age at first menses; and (2) Body Mass Index, conflicts, and sleep disturbances significantly predict the onset of puberty. Complementary standard regression analyses demonstrated that situational stress mediated the effect of these variables. We concluded that precocious puberty could be an indicator of stress in a child, thus, allowing professionals to intervene early and to prevent later problems in psychosocial adjustment.


Risk of acute and overuse injuries in youth elite soccer players: Body size and growth matter.
This study investigated anthropometric measures and growth as risk factors for overuse and acute injuries in younger (U10-U12) and older (U13-U15) elite level soccer players. Prospective cohort study. Height, weight, and sitting height were measured at the start and the end of the 2016-2017 competitive season and growth velocities were calculated. Throughout the season, injuries were registered continuously by the (para-)medical staff of the included clubs. We analyzed the injury risk using multilevel Poisson regression models, accounting for club and team clustering. Of the included 314 players (11.7±1.7 years of age), 160 players sustained 133 overuse and 163 acute injuries (i.e. 106 injuries in 69 players of the younger group, 190 in 91 players of the older group). In the younger group, risk of overuse injuries was associated with an increase in leg length over the season (incidence rate ratio (IRR) 1.620 [95% CI 1.230-2.117]) and risk of acute injuries with relatively younger age (IRR 1.003 [95% CI 1.000-1.006]). In the older group, a higher leg length was associated with an increased risk of overuse injuries (IRR 1.055 [95% CI 1.011-1.108]), and a higher weight and a lower growth rate with an increased risk of acute injuries (IRR 1.043 [95% CI 1.021-1.067] and 0.903 [95% CI 0.831-0.981], respectively). Injury risk factors differ by age group and type of injury. The age-specific anthropometric and growth-related risk factors should be monitored and these risk profiles should be considered to manage injury risk effectively.


Metanephric adenoma diagnosed on biopsy in an infant: a case report.
Metanephric adenoma is a rare benign renal tumor of the kidney, uncommonly observed in children. It is often misdiagnosed preoperatively as a malignant neoplasm, leading to an unnecessary nephrectomy. The challenge is to make the right diagnosis preoperatively and therefore manage it with conservative surgery. We report a case of a child with metanephric adenoma who underwent nephron-sparing surgery. A renal tumor was discovered fortuitously in an 18-month-old Caucasian girl with several congenital malformations. Investigations showed a 28 × 27 × 27 mm left renal mass centrally located, well defined, nonvascularized, with no calcifications and which compressed the adjacent renal tissue. Furthermore, there were no signs of metastasis. The decision of a multidisciplinary meeting was to perform a computed tomography (CT)-scan-guided biopsy. Histologic examination concluded it was a metanephric adenoma. We performed a left open partial nephrectomy via a flank retroperitoneal incision. The final histopathological examination confirmed the diagnosis. The postoperative course was uneventful. Preoperative diagnosis of metanephric adenoma is challenging. Because of the high probability of unnecessary radical nephrectomy, preoperative biopsy can be safe and determining to guide a more conservative approach so nephron-sparing surgery can be performed.


Comparing 12-core and 20-core biopsy for prostate cancer diagnosis with transperineal MR/US fusion biopsy: assessing the effective number of systemic cores using propensity score matching.
For transperineal (TP) prostate biopsy, target biopsy for visible lesions on MRI is important, but there is no consensus of the number of systemic biopsy cores. Our study aimed to confirm the diagnostic efficiency of 20-core systemic biopsy by comparison with 12-core using propensity score matching (PSM). The 494 patients conducted the naive TP biopsy were retrospectively analyzed. There were 293 patients with 12-core biopsy and 201 patients with 20-core biopsy. PSM was performed for minimizing confounding variables, and the established effects' value was analyzed for 'index-positive or negative' clinically significant prostate cancer (csPCa) (Index means PIRADS Score ≥ 3 on multiparametric prostate MRI). At 12-core biopsy, there were 126 cases of prostate cancer (43.0%), and 97 cases of csPCa (33.1%). At 20-core biopsy, there were 91 cases (45.3%) and 63 cases (31.3%). After propensity score matching, for index-negative csPCa, the estimated odds ratio was 4.03 (95% CI 1.35-12.09, p value 0.0128), and for index-positive csPCa, the estimated odds ratio was 0.98 (95% CI 0.63-1.52, p value 0.9308). The 20-core biopsy did not show a higher detection rate for csPCa in comparison with the 12-core biopsy. However, when MRI did not show a suspicious lesion, 20-core biopsy showed higher odd ratio in comparison with 12-core biopsy. Therefore, if there is a suspicious lesion in MRI, 20-core biopsy is excessive and 12-core biopsy is sufficient. Whereas if there is no suspicious lesion in MRI, it is better to proceed with 20-core biopsy.


Influence of acquisition parameters on MV-CBCT image quality.
The production of high quality pretreatment images plays an increasing role in image-guided radiotherapy (IGRT) and adaptive radiation therapy (ART). Megavoltage cone-beam computed tomography (MV-CBCT) is the simplest solution of all the commercially available volumetric imaging systems for localization. It also suffers the most from relatively poor contrast due to the energy range of the imaging photons. Several avenues can be investigated to improve MV-CBCT image quality while maintaining an acceptable patient exposure: beam generation, detector technology, reconstruction parameters, and acquisition parameters. This article presents a study of the effects of the acquisition scan length and number of projections of a Siemens Artiste MV-CBCT system on image quality within the range provided by the manufacturer. It also discusses other aspects not related to image quality one should consider when selecting an acquisition protocol. Noise and uniformity were measured on the image of a cylindrical water phantom. Spatial resolution was measured using the same phantom half filled with water to provide a sharp water/air interface to derive the modulation transfer function (MTF). Contrast-to-noise ratio (CNR) was measured on a pelvis-shaped phantom with four inserts of different electron densities relative to water (1.043, 1.117, 1.513, and 0.459). Uniformity was independent of acquisition protocol. Noise decreased from 1.96% to 1.64% when the total number of projections was increased from 100 to 600 for a total exposure of 13.5 MU. The CNR showed a ± 5% dependence on the number of projections and 10% dependence on the scan length. However, these variations were not statistically significant. The spatial resolution was unaffected by the arc length or the sampling rate. Acquisition parameters have little to no effect on the image quality of the MV-CBCT system within the range of parameters available on the system. Considerations other than image quality, such as memory storage, acquisition speed, and individual projection image quality, speak in favor of the use of a coarse sampling rate on the short scan.


Estimation of parameters in sewage sludge by near-infrared reflectance spectroscopy (NIRS) using several regression tools.
Sewage sludge application to agricultural soils is a common practice in several countries in the European Union. Nevertheless, the application dose constitutes an essential aspect that must be taken into account in order to minimize environmental impacts. In this study, near infrared reflectance spectroscopy (NIRS) was used to estimate in sewage sludge samples several parameters related to agronomic and environmental issues, such as the contents in organic matter, nitrogen and other nutrients, metals and carbon fractions, among others. In our study (using 380 biosolid samples), two regression models were fitted: the common partial least square regression (PLSR) and the penalized signal regression (PSR). Using PLSR, NIRS became a feasible tool to estimate several parameters with good goodness of fit, such as total organic matter, total organic carbon, total nitrogen, water-soluble carbon, extractable organic carbon, fulvic acid-like carbon, electrical conductivity, Mg, Fe and Cr, among other parameters, in sewage sludge samples. For parameters such as C/N ratio, humic acid-like carbon, humification index, the percentage of humic acid-like carbon, the polymerization ratio, P, K, Cu, Pb, Zn, Ni and Hg, the performance of NIRS calibrations developed with PLSR was not sufficiently good. Nevertheless, the use of PSR provided successful calibrations for all parameters.


The role of vascular endothelial growth factor and vascular endothelial growth inhibitor in clinical outcome of traumatic brain injury.
Tumor necrosis factor superfamily-15 (TNFSF15) also known as vascular endothelial growth inhibitor (VEGI) is a cytokine that modulates anti-angiogenesis and inflammation. Vascular endothelial growth factor (VEGF) promotes angiogenesis and vascular permeability following traumatic brain injury (TBI). The balance of VEGF and VEGI may play a key role in the maintenance of vascular and immune system homeostasis in the brain. However, the dynamic changes of circulating VEGF and VEGI after traumatic brain injury (TBI) and the correlation between plasma VEGF and plasma VEGI remains obscure. In this study, we were to investigate whether circulating VEGF and VEGI can be used as prognostic markers for patients with TBI. A prospective clinical study was conducted in two neurosurgical intensive care units of Tianjin Medical University General Hospital and Tianjin Huanhu Hospital (Tianjin, China). 40 patients and 30 healthy controls were recruited. The recruited subjects were aged over 18 with randomized gender and GCS. 1mL of blood was withdrawn on 1, 4, 7, 14, and 21days after TBI. Blood samples were centrifuged at 3000rpm and the supernatants were used to measure VEGF and VEGI by ELISA kit. 1) Circulating VEGF in TBI patients was decreased on the 1st day after TBI, then climbed up on the 4th day, reaching a maximum level on the14th day after TBI, as compared to normal controls. VEGF level returned to normal level on 21th day after TBI. 2) Circulating VEGI in TBI patients was decreased on the 1st and 4th day after TBI, then climbed up on the 7th day after TBI, reaching a maximum level on 14th day after TBI, as compared to normal controls. VEGI levels declined to normal level on 21th day after TBI. 3) There was a significant positive correlation between circulating VEGF and VEGI. 4) However, TBI patients whose conditions had improved exhibited lower VEGF levels 7days after TBI when compared to TBI patients whose condition had deteriorated. Survivors exhibited higher VEGI levels 7days after TBI when compared to non-survivors. 5)TBI patients whose condition had improved exhibited higher VEGI levels when compared to TBI patients whose condition had deteriorated 21days after TBI. Patients with mild TBI exhibited higher VEGI levels than those with moderate and severe TBI 21days after TBI. 6) A lower rate of recovery and higher hospital mortality were found in patients with VEGF/VEGI ratio≥2.366 as compared to those with VEGF/VEGI ratio<2.366 7days after TBI. 1) VEGF level positively correlates with VEGI after TBI. 2) The elevation of VEGF exhibits an adverse effect from 4 to 14days after TBI while it has an advantageous effect from 14 to 21days after TBI. Increasing VEGI levels are beneficial in recovery after TBI. Controlling the ratio of VEGF/VEGI may benefit the clinical outcome following TBI.


Gluconimycin an inhibitor of pyrimidine synthesis in Bacillus subtilis.
Pyrimidine synthesis in Bacillus subtilis PCI-219 cells was the primary site of action of the antibiotic gluconimycin as indicated by its arresting the activity of aspartate carbamoyltransferase as well as accumulation of aspartic acid in cultures fortified with gluconimycin. Microbial growth and viable counts were suppressed by the antibiotic whereas glycolysis and aerobic respiration were insignificantly affected. Gluconimycin failed to induce a lytic effect on cell protoplasts while considerable amounts of substances absorbing at 260 nm were released from microbial cells treated with gluconimycin.


Multicellular dynamics on structured surfaces: Stress concentration is a key to controlling complex microtissue morphology on engineered scaffolds.
Tissue engineers have utilised a variety of three-dimensional (3D) scaffolds for controlling multicellular dynamics and the resulting tissue microstructures. In particular, cutting-edge microfabrication technologies, such as 3D bioprinting, provide increasingly complex structures. However, unpredictable microtissue detachment from scaffolds, which ruins desired tissue structures, is becoming an evident problem. To overcome this issue, we elucidated the mechanism underlying collective cellular detachment by combining a new computational simulation method with quantitative tissue-culture experiments. We first quantified the stochastic processes of cellular detachment shown by vascular smooth muscle cells on model curved scaffolds and found that microtissue morphologies vary drastically depending on cell contractility, substrate curvature, and cell-substrate adhesion strength. To explore this mechanism, we developed a new particle-based model that explicitly describes stochastic processes of multicellular dynamics, such as adhesion, rupture, and large deformation of microtissues on structured surfaces. Computational simulations using the developed model successfully reproduced characteristic detachment processes observed in experiments. Crucially, simulations revealed that cellular contractility-induced stress is locally concentrated at the cell-substrate interface, subsequently inducing a catastrophic process of collective cellular detachment, which can be suppressed by modulating cell contractility, substrate curvature, and cell-substrate adhesion. These results show that the developed computational method is useful for predicting engineered tissue dynamics as a platform for prediction-guided scaffold design. STATEMENT OF SIGNIFICANCE: Microfabrication technologies aiming to control multicellular dynamics by engineering 3D scaffolds are attracting increasing attention for modelling in cell biology and regenerative medicine. However, obtaining microtissues with the desired 3D structures is made considerably more difficult by microtissue detachments from scaffolds. This study reveals a key mechanism behind this detachment by developing a novel computational method for simulating multicellular dynamics on designed scaffolds. This method enabled us to predict microtissue dynamics on structured surfaces, based on cell mechanics, substrate geometry, and cell-substrate interaction. This study provides a platform for the physics-based design of micro-engineered scaffolds and thus contributes to prediction-guided biomaterials design in the future.


[Distribution of polymorphism in endothelial nitric oxide synthase gene in Singapore Chinese and its association with diabetic nephropathy].
To examine the distribution of intron 4 polymorphism in endothelial nitric oxide synthase (eNOS) gene in mainland Chinese and Singapore Chinese and its association with diabetic nephropathy (DN). Peripheral blood samples were collected from 237 diabetes patients with the diabetic duration longer than 10 years, 87 mainland Chinese who visited the Qilu Hospital, Shandong province, and diabetic duration and blood sugar control-matched 150 Singapore Chinese who visited the Singapore National University Hospital. DNA extraction, PCR, cloning, and sequencing were conducted. The 27 bp repeat polymorphism in intron 4 was tri allelic (a, b, and c alleles) in the Singapore Chinese while biallelic (a and b alleles) in the mainland Chinese. A third allele-allele c, with six 27 bp repeats was found. Significant difference was identified in the a allele distribution between the mainland and the Singapore Chinese (P < 0.001), but the intron 4 polymorphism didn't show any association with DN. The intron 4 polymorphism is tri allelic, and the a allele distribution is not significantly associated with DN.


Signal transducer and activator of transcription (STAT)-3 regulates microRNA gene expression in chronic lymphocytic leukemia cells.
Approximately 1,000 microRNAs (miRs) are present in the human genome; however, little is known about the regulation of miR transcription. Because miR levels are deregulated in chronic lymphocytic leukemia (CLL) and signal transducer and activator of transcription (STAT)-3 is constitutively activated in CLL, we sought to determine whether STAT3 affects the transcription of miR genes in CLL cells. We used publically available data from the ENCODE project to identify putative STAT3 binding sites in the promoters of miR genes. Then we transfected CLL cells with STAT3-shRNA or with an empty vector, and to determine which miRs are differentially expressed, we used a miR microarray approach followed by validation of the microarray results for 6 miRs using quantitative real-time polymerase chain reaction (qRT-PCR). We identified putative STAT3 binding sites in 160 promoter regions of 200 miRs, including miR-21, miR-29, and miR-155, whose levels have been reported to be upregulated in CLL. Levels of 72 miRs were downregulated (n = 63) or upregulated (n = 9). qRT-PCR confirmed the array data in 5 of 6 miRs. The presence of activated STAT3 has a profound effect on miR expression in CLL cells.


Size dependent translocation and fetal accumulation of gold nanoparticles from maternal blood in the rat.
There is evidence that nanoparticles (NP) cross epithelial and endothelial body barriers. We hypothesized that gold (Au) NP, once in the blood circulation of pregnant rats, will cross the placental barrier during pregnancy size-dependently and accumulate in the fetal organism by 1. transcellular transport across the hemochorial placenta, 2. transcellular transport across amniotic membranes 3. transport through ~20 nm wide transtrophoblastic channels in a size dependent manner. The three AuNP sizes used to test this hypothesis are either well below, or of similar size or well above the diameters of the transtrophoblastic channels. We intravenously injected monodisperse, negatively charged, radio-labelled 1.4 nm, 18 nm and 80 nm ¹⁹⁸AuNP at a mass dose of 5, 3 and 27 μg/rat, respectively, into pregnant rats on day 18 of gestation and in non-pregnant control rats and studied the biodistribution in a quantitative manner based on the radio-analysis of the stably labelled ¹⁹⁸AuNP after 24 hours. We observed significant biokinetic differences between pregnant and non-pregnant rats. AuNP fractions in the uterus of pregnant rats were at least one order of magnitude higher for each particle size roughly proportional to the enlarged size and weight of the pregnant uterus. All three sizes of ¹⁹⁸AuNP were found in the placentas and amniotic fluids with 1.4 nm AuNP fractions being two orders of magnitude higher than those of the larger AuNP on a mass base. In the fetuses, only fractions of 0.0006 (30 ng) and 0.00004 (0.1 ng) of 1.4 nm and 18 nm AuNP, respectively, were detected, but no 80 nm AuNP (<0.000004 (<0.1 ng)). These data show that no AuNP entered the fetuses from amniotic fluids within 24 hours but indicate that AuNP translocation occurs across the placental tissues either through transtrophoblastic channels and/or via transcellular processes. Our data suggest that the translocation of AuNP from maternal blood into the fetus is NP-size dependent which is due to mechanisms involving (1) transport through transtrophoblastic channels - also present in the human placenta - and/or (2) endocytotic and diffusive processes across the placental barrier.


Retroviral integrations in gene therapy trials.
γ-Retroviral and lentiviral vectors allow the permanent integration of a therapeutic transgene in target cells and have provided in the last decade a delivery platform for several successful gene therapy (GT) clinical approaches. However, the occurrence of adverse events due to insertional mutagenesis in GT treated patients poses a strong challenge to the scientific community to identify the mechanisms at the basis of vector-driven genotoxicity. Along the last decade, the study of retroviral integration sites became a fundamental tool to monitor vector-host interaction in patients overtime. This review is aimed at critically revising the data derived from insertional profiling, with a particular focus on the evidences collected from GT clinical trials. We discuss the controversies and open issues associated to the interpretation of integration site analysis during patient's follow up, with an update on the latest results derived from the use of high-throughput technologies. Finally, we provide a perspective on the future technical development and on the application of these studies to address broader biological questions, from basic virology to human hematopoiesis.


Collagen type II in the otic extracellular matrix. Effect on inner ear development.
Immunocytochemistry was used to demonstrate type II collagen distribution during normal development of the mouse inner ear and in two malformed inner ears. Patterns of inner ear abnormalities and type II collagen distribution were compared between the malformed labyrinth of a mouse mutation (disproportionate micromelia, Dmm) and otic explants exposed to the teratogenic action of an L-proline analog, L-azetidine-2-carboxylic acid (LACA). The results suggest that type II collagen is an important constituent of the developing inner ear's extracellular matrix. Disruptions of the spatial and temporal pattern of collagen type II can adversely affect morphogenesis of the inner ear. A common mechanism of action is postulated for the causation of both the genetic and teratogen-induced inner ear malformations (i.e. disruption of the secretion of collagens to the otic extracellular matrix).