Tackle mechanisms and match characteristics in women's elite football tournaments.
Several tools have been used for assessing risk situations and for gathering tackle information from international football matches for men but not for women. To analyse activities in women's football and to identify the characteristics and risk potentials of tackles. Retrospective video analysis. Video recordings of 24 representative matches from six women's top-level tournaments were analysed for tackle parameters and their risk potential. 3531 tackles were recorded. Tackles in which the tackling player came from the side and stayed on her feet accounted for nearly half of all challenges for the ball in which body contact occurred. 2.7% of all tackles were classified as risk situations, with sliding-in tackles from behind and the side having the highest risk potential. Match referees sanctioned sliding-in tackles more often than other tackles (20% v 17%, respectively). Tackle parameters did not change in the duration of a match; however, there was an increase in the number of injury risk situations and foul plays towards the end of each half. Match properties provide valuable information for a better understanding of injury situations in football. Staying on feet and jumping vertically tackle actions leading to injury were sanctioned significantly more times by the referee than those not leading to injury (p<0.001), but no such difference was seen for sliding-in tackles (previously reported to have the highest injury potential in women's football). Therefore, either the laws of the game are not adequate or match referees in women's football are not able to distinguish between sliding-in tackles leading to and those not leading to injury.


Collagen Type I Improves the Differentiation of Human Embryonic Stem Cells towards Definitive Endoderm.
Human embryonic stem cells have the ability to generate all cell types in the body and can potentially provide an unlimited source of cells for cell replacement therapy to treat degenerative diseases such as diabetes. Current differentiation protocols of human embryonic stem cells towards insulin producing beta cells focus on soluble molecules whereas the impact of cell-matrix interactions has been mainly unattended. In this study almost 500 different extracellular matrix protein combinations were screened to systemically identify extracellular matrix proteins that influence differentiation of human embryonic stem cells to the definitive endoderm lineage. The percentage of definitive endoderm cells after differentiation on collagen I and fibronectin was >85% and 65%, respectively. The cells on collagen I substrates displayed different morphology and gene expression during differentiation as assessed by time lapse studies compared to cells on the other tested substrates. Global gene expression analysis showed that cells differentiated on collagen I were largely similar to cells on fibronectin after completed differentiation. Collectively, the data suggest that collagen I induces a more rapid and consistent differentiation of stem cells to definitive endoderm. The results shed light on the importance of extracellular matrix proteins for differentiation and also points to a cost effective and easy method to improve differentiation.


Maternally transmitted non-bacterial male killer in Drosophila biauraria.
A maternally inherited, all-female trait is widely found among arthropods, which is caused by bacterial endosymbionts such as Wolbachia, Rickettsia, Spiroplasma and Cardinium We discovered a single female of Drosophila biauraria, collected from Tomakomai, Hokkaido, Japan, that produced all-female offspring. This all-female trait was maternally inherited in the iso-female line (SP12F) by backcrossing with males of a normal line (SP11-20) with a 1 : 1 sex ratio derived from the same population. The all-female trait was not affected by tetracycline treatment performed for two consecutive generations. However, the microinjection of filter-sterilized homogenate of SP12F females into SP11-20 females established all-female matrilines. Our data suggest the role of transmissible agents, most likely viruses, but not bacteria or protists, as the possible cause of the all-female phenotype, which is likely to be achieved by killing of male embryos because egg hatch rates of SP12F were nearly half those of SP11-20. This is the first report in Diptera to demonstrate a maternally inherited virus-like element as the cause of the male-killing phenotype in D. biauraria.


Differences between sound scattering by weakly scattering spheres and finite-length cylinders with applications to sound scattering by zooplankton.
A modeling study was conducted to determine the conditions under which fluidlike zooplankton of the same volume but different shapes (spherical/cylindrical) have similar or dramatically different scattering properties. Models of sound scattering by weakly scattering spheres and cylinders of finite length used in this analysis were either taken from other papers or derived and herein adapted for direct comparison over a range of conditions. The models were examined in the very low- (ka << 1, kL << 1), moderately low- (ka << 1, kL > or = 1), and high-frequency regions (ka >> 1, kL >> 1), where k is the acoustic wave number, a is the radius (spherical or cylindrical) of the body, and L is the length of the cylinders (for an elongated body with L/a = 10, "moderately low" corresponds to the range 0.1 < or = ka < or = 0.5). Straight and bent cylinder models were evaluated for broadside incidence, end-on incidence, and averages over various distributions of angle of orientation. The results show that for very low frequencies and for certain distributions of orientation angles at high frequencies, the averaged scattering by cylinders will be similar, if not identical, to the scattering by spheres of the same volume. Other orientation distributions of the cylinders at high frequencies produce markedly different results. Furthermore, over a wide range of orientation distributions the scattering by spheres is dramatically different from that of the cylinders in the moderately low-frequency region and in the Rayleigh/geometric transition region: (1) the Rayleigh to geometric scattering turning point occurs at different points for the two cases when the bodies are constrained to have the same volume and (2) the functional dependence of the scattering levels upon the volume of the bodies in the moderately low-frequency region is quite often different between the spheres and cylinders because of the fact that the scattering by the cylinders is still directional in this region. The study demonstrates that there are indeed conditions under which different shaped zooplankton of the same volume will yield similar (ensemble average) scattering levels, but generally the shape and orientation distribution of the elongated bodies must be taken into account for accurate predictions.


[A resected case of pancreatic carcinoma associated with multiple intrahepatic recurrence of hepatocellular carcinoma].
A 7 1-year-old man, diagnosed with hepatocellular carcinoma (HCC), underwent lateral segmentectomy of the liver and partial hepatectomies. Ten months after the hepatic resections, computed tomography imaging of the abdomen revealed a pancreatic tumor and multiple liver metastases of HCC. The patient underwent transcatheter arterial chemoembolization three times to manage the HCC. Subsequently, he underwent distal pancreatic resection. Although it was a non-curative surgery, the postoperative course was uneventful; he was free from cancer pain, and his diabetes improved. At the 7-month follow-up after the pancreatectomy, the patients' course remained uneventful. There have been no reports of resected cases of pancreatic carcinoma that developed during the treatment of multiple intrahepatic recurrence of HCC. In cases wherein recurrence of HCC is well controlled, aggressive treatment for pancreatic cancer can be considered.


Mapping the functional domains of the Golgi stacking factor GRASP65.
The Golgi reassembly stacking protein (GRASP) family has been implicated in the stacking of Golgi cisternae and the regulation of Golgi disassembly/reassembly during mitosis in mammalian cells. GRASP65 is a dimer that can directly link adjacent surfaces through trans-oligomerization in a mitotically regulated manner. Here we show that the N-terminal GRASP domain (amino acids 1-201) is both necessary and sufficient for dimerization and trans-oligomerization but is not mitotically regulated. The C-terminal serine/proline-rich domain (amino acids 202-446) cannot dimerize nor can it link adjacent surfaces. It does, however, confer mitotic regulation on the GRASP domain through multiple sites phosphorylated by the mitotic kinases, cdc2/B1, and the polo-like kinase. Transient expression corroborated these results by showing that the GRASP domain alone inhibited mitotic fragmentation of the Golgi apparatus.


Epithelioid hemangioendothelioma of soft tissue: a proposal for risk stratification based on 49 cases.
Epithelioid hemangioendothelioma (EHE) of soft tissue is a distinctive vascular tumor that has been variously considered a tumor of borderline malignancy and low-grade angiosarcoma. The majority of cases are associated with low mortality, but some metastasize and cause patient death. We analyzed 49 cases of EHE to determine if a method for stratifying risk for mortality could be developed. Fifty-one cases of EHE diagnosed during the period of 1989 and 2005 were retrieved. Tumors were evaluated with respect to location, size, cytologic atypia, mitotic activity, tumor cell spindling, and necrosis. Follow-up information was obtained for all cases. For actuarial analysis, disease-specific overall survival was evaluated using univariate and multivariable analysis. Most tumors occurred in adults (range, 9 to 93 y) and affected women predominantly (21 M:28 F). They developed in the head and neck (6), extremities (32), mediastinum (4), trunk (4), genitals (2), and retroperitoneum (1) and ranged in size from 0.5 to 18 cm. Clinical follow-up was obtained for all patients and ranged from 1.5 to 170 months (mean, 57.9 mo); 31 patients were alive without disease, 5 patients were alive with disease, 9 patients died of disease and 4 died of other causes. Overall 5-year disease-specific survival was 81%. Eleven patients (22%) had metastatic disease affecting lung (6), lymph node (4), liver (2), and bone, retroperitoneum, and soft tissue (1 each). Two patients had metastases to multiple sites. Treatment modality was known for 46 patients: 31 were treated surgically and 15 were treated with surgery and chemotherapy and/or radiation therapy. By univariate analysis, mitotic activity and size were associated with higher mortality (P=0.007 and 0.004, respectively). By multivariable analysis, increasing mitotic activity (P=0.00827, hazard ratio 10.03) and size (P=0.01, hazard ratio 2.26) were associated with decreased survival. Tumor site, cytologic atypia, the presence of necrosis, and tumor spindling were not significant. Those tumors with >3 mitotic figures/50 high power fields and size >3.0 cm had the worst prognosis (P=0.0002). Patients with high-risk tumors had a 5-year disease-specific survival of 59%; no patients with low-risk tumors died. In conclusion, we report that EHE can be stratified into 2 risk groups with markedly different clinical courses.


Acute coronary syndromes in human immunodeficiency virus patients: a meta-analysis investigating adverse event rates and the role of antiretroviral therapy.
Highly active antiretroviral therapy (HAART) dramatically reduces human immunodeficiency virus (HIV)-associated morbidity and mortality, but adverse effects of HAART are becoming an increasing challenge, especially in the setting of acute coronary syndromes (ACS). We thus performed a comprehensive review of studies focusing on ACS in HIV patients. MEDLINE/PubMed was systematically screened for studies reporting on ACS in HIV patients. Baseline, treatment, and outcome data were appraised and pooled with random-effect methods computing summary estimates [95% confidence intervals (CIs)]. A total of 11 studies including 2442 patients were identified, with a notably low prevalence of diabetes [10.86 (4.11, 17.60); 95% CI]. Rates of in-hospital death were 8.00% (2.8, 12.5; 95% CI), ascribable to cardiovascular events for 7.90% (2.43, 13.37; 95% CI), with 2.31% (0.60, 4.01; 95% CI) developing cardiogenic shock. At a median follow-up of 25.50 months (11.25, 42; 95% CI), no deaths were recorded, with an incidence of 9.42% of acute myocardial infarction (2.68, 16.17; 95% CI) and of 20.18% (9.84, 30.51; 95% CI) of percutaneous coronary revascularization. Moreover, pooled analysis of the studies reporting incidence of acute myocardial infarction in patients exposed to protease inhibitors showed an overall significant risk of 2.68 (odds ratio 1.89, 3.89; 95% CI). Human immunodeficiency virus patients admitted for ACS face a substantial short-term risk of death and a significant long-term risk of coronary revascularization and myocardial infarction, especially if receiving protease inhibitors.


Plasmid DNA breakage by decay of DNA-associated Auger electron emitters: approaches to analysis of experimental data.
Plasmid DNA is a popular substrate for the assay of DNA strand breakage by a variety of agents. The use of the plasmid assay relies on the assumption that individual damaging events occur at random, which allows the application of Poisson statistics. This assumption is not valid in the case of damage arising from decay of DNA-associated Auger electron emitters, since a single decay event can generate a few breaks in the same DNA strand, which is indistinguishable from a single break in the assay. The consequent analytical difficulties are overcome by considering relaxation events rather than single-strand breaks, and linearization events rather than double-strand breaks. A further consideration is that apart from damage at the site of DNA-associated decay, which is the principal interest of the analysis, some DNA damage also arises from the radiation field created by all decay events. These two components of damage are referred to as internal and external breakage, respectively, and they can be separated in the analysis since their contribution depends on the experimental conditions. The DNA-binding ligand Hoechst 33258 labeled with 125I was used in our experiments to study breakage in pBR322 plasmid DNA arising from the decay of this Auger electron emitter. The values obtained for the efficiency (per decay) of plasmid relaxation and linearization by the 125I-labeled ligand were 0.090 +/- 0.035 and 0.82 +/- 0.04, respectively. When dimethylsulfoxide was included as a radical scavenger, the efficiency values for relaxation and linearization were 0.15 +/- 0.02 and 0.65 +/- 0.05, respectively.


Effects of interferon-gamma and tumor necrosis factor alpha on the expression of the genes encoding aggrecan, biglycan, and decorin core proteins in cultured human chondrocytes.
To determine the effects of interferon-gamma (IFN gamma) and tumor necrosis factor alpha (TNF alpha), alone or in combination, on the expression of aggrecan, biglycan, and decorin core protein genes in human chondrocytes. Isolated human chondrocytes were cultured on poly(2-hydroxyethyl methacrylate)-coated plastic dishes to prevent the loss of cartilage-specific phenotype, and the effects of IFN gamma and TNF alpha, alone or in combination, on aggrecan, biglycan, and decorin core protein gene transcription and steady-state messenger RNA (mRNA) levels were examined. The addition of IFN gamma (1.5 pM) or TNF alpha (0.3 pM) caused a decrease in the steady-state level of aggrecan mRNA (-25% and -15%, respectively), and the combination of these low-concentration cytokines caused a potent inhibition (-66%). These effects were the result of a decrease (-50%) in the transcription rate of the corresponding gene. At the concentrations used, IFN gamma did not alter the levels of biglycan mRNA or the transcription rates of the biglycan core protein gene. In contrast, TNF alpha decreased biglycan steady-state mRNA levels (-62%) and the biglycan core protein gene transcription rate (-18%). The combination of IFN gamma and TNF alpha resulted in a potentiation of the inhibitory effects of TNF alpha on biglycan mRNA levels (-79%) and transcription rate of the biglycan core protein gene (-46%). IFN gamma produced a modest decrease in decorin mRNA levels (-23%) and decorin core protein gene transcription rate (-17%). In contrast, TNF alpha resulted in a marked increase in decorin mRNA levels (+260%) that was not the result of transcriptional regulation. Notably, the combination of IFN gamma and TNF alpha potentiated the inhibitory effects of IFN gamma on decorin mRNA (-80%) and on the transcription of the corresponding gene (-43%). Similar results were obtained in fetal and adult articular chondrocytes. These data demonstrate that 1) the expression of the core protein genes encoding the cartilage proteoglycans aggrecan, biglycan, and decorin is differentially regulated by IFN gamma and TNF alpha; 2) these effects are mediated by transcriptional and posttranscriptional mechanisms; and 3) the combination of the 2 cytokines causes a potent inhibitory effect on the expression of the genes for the core proteins of these 3 proteoglycans, which occurs largely at the transcriptional level. The inhibition of aggrecan, decorin, and biglycan core protein gene expression by the combination of IFN gamma and TNF alpha may contribute to the cartilage destruction that is characteristic of inflammatory joint diseases.


Laparoscopic total mesorectal excision following long course chemoradiotherapy for locally advanced rectal cancer.
Laparoscopic total mesorectal excision (TME) of locally advanced rectal cancer after long-course chemoradiotherapy (LCRT) is surgically and oncologically challenging. We have assessed the feasibility, timing, and short-term oncological outcome of laparoscopic TME after LCRT. Between 2004 and 2006, 30 patients were selected for LCRT based on clinical examination and MRI. Patients received 3/4 field radiotherapy, 45-50.4 Gy in 25-28 fractions during 5 weeks with either 5-fluorouracil or Uftoral. Clinical assessments were made 4 weeks after completion of radiotherapy and then 2 weekly with sequential 4 weekly MRI, to individualize the timing of surgery at maximal response. Laparoscopic TME was performed using a standard technique. Thirty patients received LCRT and 26 patients (21 men; median age, 63 years) underwent laparoscopic TME at 11 weeks (median) after LCRT. Median operating time was 270 min. Sixteen patients had LAR and ten had APR. There were three conversions. Three patients developed anastomotic leak (18.7%): one was managed conservatively and one patient died of septicemia. Morbidity was seen in 19% of patients. There were 25 (96%) R0 resections with a complete response in 5 (19%) cases and microscopic tumor in lakes of mucin (Tmic) in another 6 (23%). Two patients (7.6%) developed local recurrence (median follow up, 34 months). The median time interval between radiotherapy and surgery was 11 (range, 7-13) weeks, which was based on serial MRI scans after LCRT. Laparoscopic TME after LCRT is feasible and safe both oncologically and surgically. Serial MRI helps to determine the optimum timing of surgery.


Noncleavable poly(ADP-ribose) polymerase-1 regulates the inflammation response in mice.
Poly(ADP-ribosyl)ation is rapidly formed in cells following DNA damage and is regulated by poly(ADP-ribose) polymerase-1 (PARP-1). PARP-1 is known to be involved in various cellular processes, such as DNA repair, genomic stability, transcription, and cell death. During apoptosis, PARP-1 is cleaved by caspases to generate 89-kDa and 24-kDa fragments, a hallmark of apoptosis. This cleavage is thought to be a regulatory event for cellular death. In order to understand the biological significance of PARP-1 cleavage, we generated a PARP-1 knockin (PARP-1(KI/KI)) mouse model, in which the caspase cleavage site of PARP-1, DEVD(214), was mutated to render the protein resistant to caspases during apoptosis. While PARP-1(KI/KI) mice developed normally, they were highly resistant to endotoxic shock and to intestinal and renal ischemia-reperfusions, which were associated with reduced inflammatory responses in the target tissues and cells due to the compromised production of specific inflammatory mediators. Despite normal binding of NF-kappaB to DNA, NF-kappaB-mediated transcription activity was impaired in the presence of caspase-resistant PARP-1. This study provides a novel insight into the function of PARP-1 in inflammation and ischemia-related pathophysiologies.


Cultured human periosteal-derived cells have inducible adipogenic activity and can also differentiate into osteoblasts in a perioxisome proliferator-activated receptor-mediated fashion.
We investigated the adipogenic activity of cultured human periosteal-derived cells and studied perioxisome proliferator-activated receptor (PPAR) ligand-mediated differentiation of cultured human periosteal-derived cells into osteoblasts. Periosteal-derived cells expressed adipogenic markers, including CCAAT/enhancer binding protein α (C/EBP- α), C/EBP-δ, aP2, leptin, LPL, and PPARγ. Lipid vesicles were formed in the cytoplasm of periosteal-derived cells. Thus, periosteal-derived cells have potential adipogenic activity. The PPARα and PPARγ agonists, WY14643 and pioglitazone, respectively, did not modulate alkaline phosphatase (ALP) activity in periosteal-derived cells during induced osteoblastic differentiation, however, the PPARα and PPARγ antagonists, GW6471 and T0070907, respectively, both decreased ALP activity in these cells. WY14643 did not affect, whereas pioglitazone enhanced, alizarin red-positive mineralization and calcium content in the periosteal-derived cells. GW6471 and T0070907 both decreased mineralization and calcium content. By RT-PCR, pioglitazone significantly increased ALP expression in periosteal-derived cells between culture day 3 and 2 weeks. Pioglitazone increased Runx2 expression after 3 days, which declined thereafter, but did not alter osteocalcin expression. Both of GW6471 and T0070907 decreased ALP mRNA expression. These results suggest that pioglitazone enhances osteoblastic differentiation of periosteal-derived cells by increasing Runx2 and ALP mRNA expression, and increasing mineralization. GW6471 and T0070907 inhibit osteoblastic differentiation of the periosteal-derived cells by decreasing ALP expression and mineralization in the periosteal-derived cells. In conclusion, although further study will be needed to clarify the mechanisms of PPAR-regulated osteogenesis, our results suggest that PPARγ agonist stimulates osteoblastic differentiation of cultured human periosteal-derived cells and PPARα and PPARγ antagonists inhibit osteoblastic differentiation in these cells.


Effects of recent word exposure on emotion-word Stroop interference: an ERP study.
Attentional bias towards emotional linguistic material has been examined extensively with the emotion-word Stroop task. Although findings in clinical groups show an interference effect of emotional words that relate to the specific concern of the group, findings concerning healthy groups are less clear. In the present study, we investigated whether emotional Stroop interference in healthy individuals is affected by exposure of the words prior to the task. We used event-related potentials (ERPs) to examine the temporal aspects of Stroop interference. Participants took longer to indicate the colour of negative than of neutral words. Exposure of words prior to the Stroop task increased response latencies, but this effect was equal for neutral and negative words. At the neurophysiological level, we found more positive-going ERPs at later latencies (P290, N400 and LPP) in response to negative than in response to neutral Stroop words. The N400 was less negative for exposed than for new words, but this effect did not interact with the emotional valence of the words. For new (i.e., unexposed) words, the behavioural Stroop interference correlated with the P290, N400 and LPP emotion effects (negative minus neutral words). The successive ERP components suggest better prelexical, semantic, and sustained attentional processing of emotion words, even when the emotional content of the words is task-irrelevant.


Rate and Reasons for Repeat CT Scanning in Transferred Trauma Patients.
To describe the reasons and additional radiation for CT scans repeated after transfer to a trauma center. Retrospective study of patients transferred to a trauma center. Patients had repeat CT if a CT of the same region was repeated at the trauma center's emergency department. Indications for repeat CT scanning were abstracted. Radiation dosage was calculated in millisieverts. A total of 370 had CT scans and were transferred. Mean age was 39.1 ± 28.0 years. Seventy-four [20.0%, 95% confidence interval (CI) 16.0-24.4%] had 103 CTs repeated. Adults (64/254, 25.2%) were more likely than children (10/116, 8.6%) to undergo repeat CT (difference 16.6%, 95% CI 9.2-24.0%). Types of CTs repeated included: head 48 (47%), face 6 (6%), cervical spine/neck 21 (20%), thoracolumbar spine 4 (4%), chest 4 (4%), and abdominal/pelvic 20 (19%). Reasons for repeat CT were outside CT unavailable 31 (42%), insufficient image quality/additional details needed 15 (20%), disease progression 16 (22%), unknown 10 (14%), and consult request unknown reason 2 (3%). Median dose for the repeat CT scans was 4.19 mSv (interquartile range 1.98, 6.28) and was 4.79 mSv (interquartile range 2.47, 8.22) when the CTs were unavailable. Effective dose of the repeat scans was greater than 10 mSv in 13 (3.5%) patients. Patients transferred to a trauma center often undergo repeat CT. The most common reason for repeated imaging was failure to transport original CT scans with the patient or images that were unable to be viewed. Trauma centers should work with their catchment areas to establish systems that ensure transfer of all radiographic imaging.


Dementia and aggressiveness: stimulated recall interviews with caregivers after video-recorded interactions.
In a previous study, nine caregivers and two residents with dementia showing aggressive behaviour, were video recorded. Caregivers who reported problems when dealing with such behaviour and caregivers, who did not, were included in this study. The aim of the present study was to obtain insight into the reasoning of the caregivers who had reported problems when dealing with older people with dementia and aggressiveness and those who did not relative to their respective video-recorded interactions with these residents. A further aim was to gain insight by discussing their reasoning in relation to each other. Stimulated recall interviews were carried out with all the caregivers who had been video taped in the previous study. The text was analysed by thematic content analysis. Two main ways of thinking and discussing the care situations emerged. The caregivers, who had reported problems in handling behavioural and psychiatric symptoms in dementia earlier, reasoned that they were more focused on their duties, this included being responsible for the resident receiving her weekly shower. For this group of caregivers, the well being of the resident was in focus, but their attention was concentrated on the resident's well being and comfort after their shower. However, these caregivers seemed therefore unwittingly to prevent a positive interaction with the resident. The other caregivers were able to reflect spontaneously and appeared to be self-critical. This caregiver group seemed to sustain a positive interaction with the resident both during and after the shower. In this study a nurturing and supportive climate and competence seemed to be the conditions necessary to facilitate reflections and promote creativity in the caregivers such that they are able to develop possible ways of handling difficult situations like aggressiveness in residents with dementia.


Conformational properties of bacterial DnaK and yeast mitochondrial Hsp70. Role of the divergent C-terminal alpha-helical subdomain.
Among the eukaryotic members of the Hsp70 family, mitochondrial Hsp70 shows the highest degree of sequence identity with bacterial DnaK. Although they share a functional mechanism and homologous co-chaperones, they are highly specific and cannot be exchanged between Escherichia coli and yeast mitochondria. To provide a structural basis for this finding, we characterized both proteins, as well as two DnaK/mtHsp70 chimeras constructed by domain swapping, using biochemical and biophysical methods. Here, we show that DnaK and mtHsp70 display different conformational and biochemical properties. Replacing different regions of the DnaK peptide-binding domain with those of mtHsp70 results in chimeric proteins that: (a) are not able to support growth of an E. coli DnaK deletion strain at stress temperatures (e.g. 42 degrees C); (b) show increased accessibility and decreased thermal stability of the peptide-binding pocket; and (c) have reduced activation by bacterial, but not mitochondrial co-chaperones, as compared with DnaK. Importantly, swapping the C-terminal alpha-helical subdomain promotes a conformational change in the chimeras to an mtHsp70-like conformation. Thus, interaction with bacterial co-chaperones correlates well with the conformation that natural and chimeric Hsp70s adopt in solution. Our results support the hypothesis that a specific protein structure might regulate the interaction of Hsp70s with particular components of the cellular machinery, such as Tim44, so that they perform specific functions.


Effects of inhibition of microtubule assembly on bone mineral release and enzyme release by human breast cancer cells.
When supernates from the established human breast cancer cell line MCF-7 were applied to fetal rat long bones that had been labeled with 45Ca and devitalized to remove endogenous bone cells, mineral was released from the bones. The release of bone mineral by MCF-7 supernates was associated with increased basal release of hydrolytic enzyme activity by the tumor cells. The basal release of lysosomal enzymes and collagenolytic activity by MCF-7 cells with approximately twice that of mouse 3T3 cells, which did not cause mineral release by the fetal rat bones. Release of hydrolytic enzymes and bone mineral-releasing activity was increased by colchicine and vinblastine, drugs that inhibit microtubule assembly, but not affected by lumicolchicine. Time-course experiments performed on MCF-7 cells with or without colchicine showed that release of cathepsin D and collagenolytic activity was associated more closely with release of bone mineral and degradation of bone matrix than was the release of N-acetylglucosaminidase. The release of previously incorporated [3H]proline from the bones exposed to MCF-7 cell cultures was more closely associated with release of collagenolytic activity by MCF-7 cells than with release of cathepsin D or N-acetylglucosaminidase. These data suggest that breast cancer-mediated bone resorption in vitro is positively correlated with release of hydrolytic enzymes by the tumor cells, and release of these enzymes is enhanced by disassembly of microtubules.


Prenatal exposure to a novel antipsychotic quetiapine: impact on neuro-architecture, apoptotic neurodegeneration in fetal hippocampus and cognitive impairment in young rats.
Reports on prenatal exposure to some of the first generation antipsychotic drugs like, haloperidol, their effects on fetal neurotoxicity and functional impairments in the offspring, are well documented. But studies on in utero exposure to second generation antipsychotics, especially quetiapine, and its effects on fetal neurotoxicity, apoptotic neurodegeneration, postnatal developmental delay and neurobehavioral consequences are lacking. Therefore, the present study was undertaken to evaluate the effect of prenatal administration to equivalent therapeutic doses of quetiapine on neuro-architectural abnormalities, neurohistopathological changes, apoptotic neurodegeneration in fetal hippocampus, and postnatal development and growth as well as its long-lasting imprint on cognitive impairment in young-adult offspring. Pregnant Wistar rats (n=24) were exposed to selected doses (55 mg, 80 mg and 100mg/kg) of quetiapine, equivalent to human therapeutic doses, from gestation day 6 to 21 orally with control subjects. Half of the pregnant subjects of each group were sacrificed at gestation day 21 for histopathological, confocal and electron microscopic studies and rest of the dams were allowed to deliver naturally. Their pups were reared postnatally up to 10 weeks of age for neurobehavioral observations. In quetiapine treated groups, there was significant alterations in total and differential thickness of three typical layers of hippocampus associated with neuronal cells deficit and enhanced apoptotic neurodegeneration in the CA1 area of fetal hippocampus. Prenatally drug treated rat offspring displayed post-natal developmental delay till postnatal day 70, and these young-adult rats displayed cognitive impairment in Morris water maze and passive avoidance regimes as long-lasting impact of the drug. Therefore, quetiapine should be used with cautions considering its developmental neurotoxicological and neurobehavioral potential in animal model, rat.


[Takotsubo cardiomyopathy associated with abnormal birth of the coronary arteries].
Takotsubo cardiomyopathy (TM) is a form of non-ischemic cardiomyopathy. It is characterized by transient regional systolic dysfunction of the left ventricle that mimics acute myocardial infarction. The main objective of this article is to report the case of a patient with TM associated with abnormal birth of the left coronary trunk. A 76-year-old woman with typical angina at rest, with an electrocardiogram that showed dynamic changes in the T wave and elevation of biomarkers. Coronary angiography showed epicardial coronary arteries without significant lesions and abnormal birth of the left coronary trunk from the proximal segment of the right coronary artery. The patient progressed favorably, and the transthoracic echocardiogram showed no mobility disorders 3 months after the event. TM and abnormal birth of the coronary arteries are rare diseases whose simultaneous presentation is extraordinary. The diagnosis of both clinical entities is made by coronary angiography and echocardiogram, and their treatment is similar to that of patients with acute coronary syndrome. Abnormal birth of the left coronary trunk with retroaortic switch reaching the contralateral site has a good clinical prognosis and echocardiographic follow-up should be performed 4 weeks after the onset of the condition.


Lentiviral hematopoietic stem cell gene therapy for X-linked severe combined immunodeficiency.
X-linked severe combined immunodeficiency (SCID-X1) is a profound deficiency of T, B, and natural killer (NK) cell immunity caused by mutations inIL2RGencoding the common chain (γc) of several interleukin receptors. Gamma-retroviral (γRV) gene therapy of SCID-X1 infants without conditioning restores T cell immunity without B or NK cell correction, but similar treatment fails in older SCID-X1 children. We used a lentiviral gene therapy approach to treat five SCID-X1 patients with persistent immune dysfunction despite haploidentical hematopoietic stem cell (HSC) transplant in infancy. Follow-up data from two older patients demonstrate that lentiviral vector γc transduced autologous HSC gene therapy after nonmyeloablative busulfan conditioning achieves selective expansion of gene-marked T, NK, and B cells, which is associated with sustained restoration of humoral responses to immunization and clinical improvement at 2 to 3 years after treatment. Similar gene marking levels have been achieved in three younger patients, albeit with only 6 to 9 months of follow-up. Lentiviral gene therapy with reduced-intensity conditioning appears safe and can restore humoral immune function to posthaploidentical transplant older patients with SCID-X1.


The cerebellotectal pathway in the grey squirrel.
In the well laminated superior colliculus of the grey squirrel the cells of origin of the crossed descending pathway to the brainstem gaze centers are contained within the inner sublamina of the intermediate grey layer. The technique of anterograde transport of horseradish peroxidase was used to determine whether the pathway from the cerebellum to the superior colliculus terminates in this region. The technique of retrograde transport of horseradish peroxidase was used to localize the source of this pathway within the cerebellum and to determine the morphology of the cerebellotectal neurons. The grey squirrel cerebellotectal pathway provides two terminal fields to the superior colliculus: a diffuse projection into the deep grey layer and a more concentrated, interrupted projection into the inner sublamina of the intermediate grey layer. The more concentrated projection overlies precisely the tectal sublamina that contains the cells of origin of the predorsal bundle. In contrast to animals with frontal eyes, the cerebellotectal pathway in the grey squirrel was found to project almost entirely contralaterally and the vast majority of the cells of origin for the pathway were distributed ventrally, in the caudal pole of the posterior interpositus nucleus and the adjacent region of the dentate. The labelled cells in both cerebellar nuclei were large and displayed similar morphologies.


Risk factors for positive depression screening across a shipboard deployment cycle.
Depression is a leading cause of healthcare use and risk for suicide among US military personnel. Depression is not well characterised over the shipboard deployment cycle, and personnel undergo less screening than with land-based deployments, making early identification less likely. To determine the demographic and behavioural risk factors associated with screening positive for risk of depression (ROD) across the shipboard deployment cycle. Active-duty ship assigned personnel completed an anonymous assessment using the Center for Epidemiologic Studies Depression Scale (CES-D) in the year prior to deployment, during deployment and in the months following deployment. Longitudinal models were used to determine risk factors. In total, 598 people were included in the analysis. Over 50% of the study population screened positive for ROD (CES-D score ≥16) and over 25% screened positive for risk of major depressive disorder (CES-D score ≥22) at all time points. Lower age, female gender, alcohol use, stress and prior mental health diagnoses were all associated with greater odds of screening positive for ROD in multivariable models. Although the risk factors associated with screening positive for ROD are similar to those in other military and civilian populations, the proportion screening positive exceeds previously reported prevalence. This suggests that shipboard deployment or factors associated with shipboard deployment may present particular stressors or increase the likelihood of depressive symptoms. The authors are military service members (or employees of the US Government). This work was prepared as part of the authors' official duties. Title 17, U.S.C. §105 provides the 'Copyright protection under this title is not available for any work of the United States Government.' Title 17, USC, §101 defines a US Government work as work prepared by a military service member or employee of the US Government as part of that person's official duties. The views expressed in this research are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Department of the Army, Department of the Air Force, Department of Veterans Affairs, Department of Defense, or the US Government. Approved for public release; distribution unlimited. Material has been reviewed by the Walter Reed Army Institute of Research. There is no objection to its presentation and/or publication. The opinions or assertions contained herein are the private views of the authors, and are not to be construed as official, or as reflecting true views of the Department of the Army or the Department of Defense.


Robotic-assisted surgery for primary or recurrent oropharyngeal carcinoma.
To determine the feasibility of robotic-assisted salvage surgery for oropharyngeal cancer. Retrospective case-controlled study. Academic, tertiary referral center. Patients who underwent surgical resection for T1 and T2 oropharyngeal cancer between 2001 and 2008 were classified into the following 3 groups based on type of resection: (1) robotic-assisted surgery for primary neoplasms (robotic primary) (n = 15), (2) robotic-assisted salvage surgery for recurrent disease (robotic salvage) (n = 7), and (3) open salvage resection for recurrent disease (n = 14). Data regarding tumor subsite, stage, and prior treatment were evaluated as well as margin status, nodal disease, length of hospital stay, diet, and tracheotomy tube dependence. The median length of stay in the open salvage group was longer (8.2 days) than robotic salvage (5.0 days) (P = .14) and robotic primary (1.5 days) resection groups (P < .001). There was no difference in postoperative diet between robotic primary and robotic salvage surgery groups. However, a greater proportion of patients who underwent open salvage procedures were gastrostomy tube dependent 6 months following treatment (43%) compared with robotic salvage resection (0%) (P = .06). A greater proportion of patients who underwent open salvage procedures also remained tracheotomy tube dependent after 6 months (7%) compared with robotic salvage or robotic primary patients (0%) (P = .48). No complications were reported in the robotic salvage group. Two patients who underwent open salvage resection developed postoperative hematomas and 2 developed wound infections. When feasible, robotic-assisted surgery is an acceptable procedure for resection of both primary and recurrent oropharyngeal tumors. Trial Registration clinicaltrials.gov Identifier: NCT00473564.


[The Healthcare Teams' Perspective on Caring for People with Dementia in Acute Hospitals: A Qualitative Study].
The Healthcare Teams' Perspective on Caring for People with Dementia in Acute Hospitals: A Qualitative Study Abstract. Introduction: To deal with the complexity of the situation of people with dementia in acute hospitals, it is necessary to develop tailored interventions. In doing so, it is important to consider the perspectives of all relevant persons, including health care teams. The aim of this study was to explore the situation of people with dementia in three Swiss acute hospitals from the perspective of health care teams. We conducted three focus group interviews with health care teams consisting of medical doctors, nurses and therapists. Data were analysed by means of summarising content analysis according to Mayring. A total of 20 health professionals took part. Three main categories were identified: "People with dementia confront the hospital system", "The hospital system fails to meet the needs of people with dementia" and "Necessary changes take place in the hospital system". The results show a lack of intention in the hospital system to address the specific needs of people with dementia. Health care teams feel forced to intensify their teamwork. This occurs unsystematically and with little organisational support. It seems of paramount importance to systematically support the teams' initiatives for enhanced teamwork in caring for people with dementia. Teamwork should be considered as a key aspect when developing interventions.


Characterization and expression analysis of a serine acetyltransferase gene family involved in a key step of the sulfur assimilation pathway in Arabidopsis.
Ser acetyltransferase (SATase; EC 22.214.171.124) catalyzes the formation of O-acetyl-Ser from L-Ser and acetyl-CoA, leading to synthesis of Cys. According to its position at the decisive junction of the pathways of sulfur assimilation and amino acid metabolism, SATases are subject to regulatory mechanisms to control the flux of Cys synthesis. In Arabidopsis (Arabidopsis thaliana) there are five genes encoding SATase-like proteins. Two isoforms, Serat3;1 and Serat3;2, were characterized with respect to their enzymatic properties, feedback inhibition by L-Cys, and subcellular localization. Functional identity of Serat3;1 and Serat3;2 was established by complementation of a SATase-deficient mutant of Escherichia coli. Cytosolic localization of Serat3;1 and Serat3;2 was confirmed by using fusion construct with the green fluorescent protein. Recombinant Serat3;1 was not inhibited by L-Cys, while Serat3;2 was a strongly feedback-inhibited isoform. Quantification of expression patterns indicated that Serat2;1 is the dominant form expressed in most tissues examined, followed by Serat1;1 and Serat2;2. Although Serat3;1 and Serat3;2 were expressed weakly in most tissues, Serat3;2 expression was significantly induced under sulfur deficiency and cadmium stress as well as during generative developmental stages, implying that Serat3;1 and Serat3;2 have specific roles when plants are subjected to distinct conditions. Transgenic Arabidopsis plants expressing the green fluorescent protein under the control of the five promoters indicated that, in all Serat genes, the expression was predominantly localized in the vascular system, notably in the phloem. These results demonstrate that Arabidopsis employs a complex array of compartment-specific SATase isoforms with distinct enzymatic properties and expression patterns to ensure the provision of Cys in response to developmental and environmental changes.


CD36/fatty acid translocase, an inflammatory mediator, is involved in hyperlipidemia-induced exacerbation in ischemic brain injury.
Hyperlipidemia with accompanying increase in peripheral inflammation is a risk factor for stroke. The effect of excess lipids on stroke-induced injury and the mechanism by which lipid-mediated inflammatory responses contribute to stroke are not known. We investigated these uncertainties by subjecting normal and hyperlipidemic mice to transient middle cerebral artery occlusion, followed by measurement of stroke severity and inflammatory response. Infarct size, swelling, and lipid contents were significantly increased in the high-fat fed ApoE knock-out mice, as was the expression of the inflammatory mediators CD36 and monocyte chemoattractant protein 1 (MCP-1) in the brain and periphery. Furthermore, the hyperlipidemic mice exhibited numerous foam cells, a probable cause of increased swelling and postischemic inflammation, in the peri-infarct area. Genetic deletion of cd36 in the hyperlipidemic condition reduced proinflammatory chemokine/receptor and cytokines (MCP-1, CC chemokine receptor 2, and interleukins 1beta and 6), in the brain 6 h after ischemia. The reduced proinflammatory response also resulted in smaller ischemic injury, less swelling, and fewer foam cells at 3 d after ischemia. The results show that hyperlipidemia-induced inflammation is a negative factor for stroke outcomes and indicate that downregulating CD36 may be an effective therapeutic strategy for reducing the impact of stroke in hyperlipidemic subjects.


Child and adolescent mental health in South Africa.
Mental health problems in childhood and adolescence pose a major threat to public health. Epidemiological studies in high, middle and low income countries indicate that approximately one in five children and adolescents suffer from a mental disorder. In many instances these persist into adulthood. In South Africa, HIV infection, substance use, and exposure to violence increase vulnerability to mental disorders. Child and adolescent mental health services play a key role in reducing the burden of mental disorders in childhood and later in adulthood. This paper focuses on service needs for children and adolescents in South Africa. It commences with a discussion of the prevalence of child and adolescent psychiatric disorders after which the legal and policy context of child and adolescent psychiatric services is described. A framework for child and adolescent mental health service provision is presented, following which steps for reducing the extent of unmet service need are considered. The paper concludes with a call to scale up child and adolescent mental health services in South Africa, based on the stark realities of unmet need and the constitutional rights of children and adolescents to appropriate mental health care.


Chlorogenic acid induces apoptosis, inhibits metastasis and improves antitumor immunity in breast cancer via the NF‑κB signaling pathway.
Breast cancer which is the most common type of diagnosed cancer among women worldwide possesses metastatic potential, multi‑drug resistance, and high mortality. The NF‑κB signaling pathway has been revealed to be abnormally activated in breast cancer cells and closely associated with high metastasis and poor prognosis. In the present study, it was reported that chlorogenic acid (CGA), a potent NF‑κB inhibitor derived from coffee, exerted antitumor activity in breast cancer. MTT and colony formation assays were conducted and it was revealed that CGA inhibited viability and proliferation in breast cancer cells. Additionally, CGA significantly induced apoptosis and suppressed migration and invasion in breast cancer cells. Notably, immunofluorescence analysis confirmed that CGA could efficiently suppress nuclear transcription of NF‑κB p65. In addition, results of western blotting demonstrated that CGA markedly impaired the NF‑κB and EMT signaling pathways. The antitumor effect of CGA was evaluated in a subcutaneous tumor mouse model of 4T1 cells, and the results revealed that CGA markedly retarded tumor growth and prolonged the survival rate of tumor‑bearing mice. Notably, CGA inhibited pulmonary metastasis of 4T1 cells by enhancing the proportion of CD4+ and CD8+ T cells in spleens of mice, which indicated an improvement of antitumor immunity. In conclusion, the present present study demonstrated that CGA improved antitumor immunity, exerting antitumor and anti‑metastatic effects by impairing the NF‑κB/EMT signaling pathway, suggesting that CGA may serve as a potential candidate for therapy of breast cancer.


[Differentiation of endocrinocytes in the small-intestinal epithelium of albino rats in ontogenesis].
By means of the light microscopy method differentiation and dynamics on contents of endocrinocytes have been studied in epithelium of the initial part of the jejunum, of the caudal part of the ileum and of the middle part between them during pre- and postnatal periods of development. Formation of the endocrine apparatus of the mucous membrane epithelium in the small intestine takes place simultaneously with tissue differentiation of the epithelial layer. The population density of endotheliocytes of the mucous membrane is maximal in the initial part of the jejunum and decreases gradually in the caudal direction. By the 15th day of the postnatal development endocrinocytes in composition of the epithelium in the small intestine reach a high level of the specific differentiation and their amount increases by 1.3 times in comparison with that in newborn animals.


Hsp40 interacts directly with the native state of the yeast prion protein Ure2 and inhibits formation of amyloid-like fibrils.
Ure2 is the protein determinant of the [URE3] prion phenotype in Saccharomyces cerevisiae and consists of a flexible N-terminal prion-determining domain and a globular C-terminal glutathione transferase-like domain. Overexpression of the type I Hsp40 member Ydj1 in yeast cells has been found to result in the loss of [URE3]. However, the mechanism of prion curing by Ydj1 remains unclear. Here we tested the effect of overexpression of Hsp40 members Ydj1, Sis1, and Apj1 and also Hsp70 co-chaperones Cpr7, Cns1, Sti1, and Fes1 in vivo and found that only Ydj1 showed a strong curing effect on [URE3]. We also investigated the interaction of Ydj1 with Ure2 in vitro. We found that Ydj1 was able to suppress formation of amyloid-like fibrils of Ure2 by delaying the process of fibril formation, as monitored by thioflavin T binding and atomic force microscopy imaging. Controls using bovine serum albumin, Sis1, or the human Hsp40 homologues Hdj1 or Hdj2 showed no significant inhibitory effect. Ydj1 was only effective when added during the lag phase of fibril formation, suggesting that it interacts with Ure2 at an early stage in fibril formation and delays the nucleation process. Using surface plasmon resonance and size exclusion chromatography, we demonstrated a direct interaction between Ydj1 and both wild type and N-terminally truncated Ure2. In contrast, Hdj2, which did not suppress fibril formation, did not show this interaction. The results suggest that Ydj1 inhibits Ure2 fibril formation by binding to the native state of Ure2, thus delaying the onset of oligomerization.


Endogenous secretory receptor for advanced glycation end products is associated with low serum interleukin-1 receptor antagonist and elevated IL-6 in older community-dwelling adults.
Advanced glycation end products (AGEs) are thought to cause inflammation through interaction with the receptor for AGEs (RAGE), therefore contributing to adverse aging-related processes. The relationship between AGEs, RAGE, and inflammation has not been well characterized. We examined the relationship of plasma endogenous secretory RAGE (esRAGE); carboxymethyl-lysine (CML), a circulating AGE; and inflammatory mediators in 1,298 adults, 20-97 years, who participated in the InCHIANTI study in Tuscany, Italy. Blood levels of esRAGE, CML, interleukin-1 receptor antagonist (IL-1RA), IL-1β, tumor necrosis factor-α (TNF-α), IL-6, IL-6 receptor (IL-6R), IL-18, C-reactive protein (CRP), transforming growth factor-β (TGF-β), and fibrinogen were measured. Log plasma esRAGE was associated with log IL-1RA (β = -0.069, SE = 0.036, p = .05) and log IL-6 (β = 0.077, SE = 0.035, p = .03), respectively, in separate multivariable linear regression models, adjusting for potential confounders. Log plasma esRAGE was also negatively associated with log TGF-β but did not reach statistical significance (β = -0.091, SE = 0.053, p = .09). Log plasma esRAGE was not significantly associated with log IL-1β, log TNF-α, IL-6R, log IL-18, or CRP. Log plasma CML was not associated with any of the inflammatory mediators except for IL-6R (β = -14.10, SE = 5.94, p = .02) and fibrinogen (β = 13.95, SE = 7.21, p = .05) in separate multivariable models, adjusting for potential confounders. Plasma esRAGE is correlated with higher IL-6 and lower IL-1RA. These findings suggest that plasma esRAGE plays a role in modulating inflammation, although the exact mechanisms remain to be elucidated.


Preoperative evaluation of pediatric kidney stone prior to percutaneous nephrolithotomy: is computed tomography really necessary?
The objective of the study was to investigate the precise role of computed tomography (CT) in preoperative radiologic evaluation and surgical planning of kidney stone in children prior to percutaneous nephrolithotomy (PNL). A total of 113 pediatric patients (aged ≤18 years) undergoing PNL for renal stone(s) in three referral hospitals between March 2010 and August 2012 were retrospectively evaluated. Depending on the preoperative radiologic evaluation, patients were divided into two groups. Those evaluated with CT were classified as group-1 (n = 50) and the remaining cases undergoing intravenous urography (IVU) examination were classified as group-2 (n = 63). Patient- and procedure-related variables and perioperative measures were compared between the groups. The mean age, stone size and localization were similar in both groups (p = 0.07, p = 0.57, p = 0.6, respectively). Although the postoperative hemoglobin drop was found to be significantly higher in group-2 (1.5 ± 1.3 vs. 0.9 ± 0.6 g/dL, p = 0.005), the mean operation time, fluoroscopic screening time, access number, overall success and complication rates were comparable (p = 0.06, p = 0.94, p = 0.75, p = 041, and p = 0.41, respectively). However, the mean hospitalization time was significantly prolonged in group-2 than in group-1 (p = 0.03). Our findings clearly demonstrate that, despite the key role of preoperative CT in particular patients with anatomically abnormal kidneys, IVU is a valuable alternative imaging modality with comparable radiation doses in children.


Photocrosslinkable polysaccharides based on chondroitin sulfate.
Photopolymerizing hydrogels have demonstrated potential for use as a scaffold in numerous tissue-engineering applications. The majority of photopolymerizing hydrogels are made from purely synthetic polymers. The purpose of this study was to synthesize and characterize photopolymerizing hydrogels derived from the biopolymer chondroitin sulfate in order to enhance the bioactivity of the scaffold and potentially improve tissue regeneration. Methacrylate groups were added to chondroitin sulfate, a major component of cartilage, using glycidyl methacrylate. The gels exhibited viscoelastic behavior typical of hydrogels. Cogels based on chondroitin sulfate and poly(ethylene glycol) demonstrated increasing pore size with increasing concentration of chondroitin sulfate as determined by water content, mechanical strength, and morphology using scanning electron microscopy. The chondroitin sulfate hydrogels degraded specifically in the presence of the enzyme chondroitinase. Chondrocytes remained viable after photoencapsulation and incubation in the biogels, suggesting their possible use for cartilage tissue engineering.


Molecular Individual-Based Approach on Triatoma brasiliensis: Inferences on Triatomine Foci, Trypanosoma cruzi Natural Infection Prevalence, Parasite Diversity and Feeding Sources.
We used an individual-based molecular multisource approach to assess the epidemiological importance of Triatoma brasiliensis collected in distinct sites and ecotopes in Rio Grande do Norte State, Brazil. In the semi-arid zones of Brazil, this blood sucking bug is the most important vector of Trypanosoma cruzi--the parasite that causes Chagas disease. First, cytochrome b (cytb) and microsatellite markers were used for inferences on the genetic structure of five populations (108 bugs). Second, we determined the natural T. cruzi infection prevalence and parasite diversity in 126 bugs by amplifying a mini-exon gene from triatomine gut contents. Third, we identified the natural feeding sources of 60 T. brasiliensis by using the blood meal content via vertebrate cytb analysis. Demographic inferences based on cytb variation indicated expansion events in some sylvatic and domiciliary populations. Microsatellite results indicated gene flow between sylvatic and anthropic (domiciliary and peridomiciliary) populations, which threatens vector control efforts because sylvatic population are uncontrollable. A high natural T. cruzi infection prevalence (52-71%) and two parasite lineages were found for the sylvatic foci, in which 68% of bugs had fed on Kerodon rupestris (Rodentia: Caviidae), highlighting it as a potential reservoir. For peridomiciliary bugs, Galea spixii (Rodentia: Caviidae) was the main mammal feeding source, which may reinforce previous concerns about the potential of this animal to link the sylvatic and domiciliary T. cruzi cycles.


Depression in patients with head and neck cancer and a functional genetic polymorphism of the serotonin transporter gene.
Polymorphism of the serotonin transporter gene, 5-HTTLPR (short allele) has been associated with depression. The purpose of this study was to show the evaluated depression in patients with head and neck cancer and a possible association with the 5-HTTLPR. The Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders (SCID for DSM-IV) was administered to 94 patients with head and neck cancer, of which 33 patients were genotyped for 5-HTTLPR. We also evaluated the prevalence of 5-HTTLPR polymorphism in 121 patients with head and neck cancer and 97 controls. Forty-three percent of the patients met the criteria for a depressive diagnosis, 19% of which was new onset. In depressed patients, 85.7% (n = 12 of 14) had at least 1 short allele versus 68.4% (n = 13 of 19) of the patients without depressive diagnosis (p < .04). No difference was noted in the prevalence of the short allele in head and neck cancer cases versus controls (odds ratio = 0.8; p = .490). Despite the high rate of depressive diagnosis, patients with head and neck cancer did not demonstrate a higher prevalence of this short allele of the 5-HTTLPR compared with a control population.


Evaluation of patient satisfaction using the validated French version of the SSIPI questionnaire after inflatable penile implantation.
Penile prothesis (PP) is a recommended treatment for erectile dysfunction that is refractory to less invasive treatments, but there are few validated tools to assess patient satisfaction. The aim of this study was to assess patient satisfaction after PP implantation using the French Satisfaction Survey for Inflatable Penile Implant (SSIPI) questionnaire. Demographic, clinical, and perioperative data were collected from all consecutive patients who underwent PP implantation in our center between 2016 and 2021. The French SSIPI questionnaire was completed during a telephone call with each patient by an independent investigator. A good functional result was considered when the total SSIPI score was >48, corresponding to a score of >3 for each item. The median global SSIPI score for the cohort was 66 (interquartile range [IQR], 60-73), and 53 (89.8%) patients were satisfied. The appearance of the penis with the PP was the item that had the lowest score for satisfaction (median score 23 [IQR, 19-26]), while the patients reported almost no pain (median score 10 [IQR, 9-10]). When patients with a total score of ≤64 were compared with those with a score of >64, PP size was significantly greater in the group with better functional results (P = .03). Assessment of patient satisfaction with a PP is important because this is the main criterion used to judge the success of surgery. To our knowledge, this is the first study to evaluate the satisfaction of patients with a PP using the French-validated version of SSIPI questionnaire. However, patients came from a single center and the population size was small. Almost 90% of patients with a PP were satisfied with the device when satisfaction was assessed using the SSIPI questionnaire.


Stimulus-dependent translocation of kappa opioid receptors to the plasma membrane.
We examined the cellular and subcellular distribution of the cloned kappa opioid receptor (KOR1) and its trafficking to the presynaptic plasma membrane in vasopressin magnocellular neurosecretory neurons. We used immunohistochemistry to show that KOR1 immunoreactivity (IR) colocalized with vasopressin-containing cell bodies, axons, and axon terminals within the posterior pituitary. Ultrastructural analysis revealed that a major fraction of KOR1-IR was associated with the membrane of peptide-containing large secretory vesicles. KOR1-IR was rarely associated with the plasma membrane in unstimulated nerve terminals within the posterior pituitary. A physiological stimulus (salt-loading) that elicits vasopressin release also caused KOR1-IR to translocate from these vesicles to the plasma membrane. After stimulation, there was a significant decrease in KOR1-IR associated with peptide-containing vesicles and a significant increase in KOR1-IR associated with the plasma membrane. This stimulus-dependent translocation of receptors to the presynaptic plasma membrane provides a novel mechanism for regulation of transmitter release.


[Structure of maxillary sinus mucous membrane under normal conditions and in odontogenic perforative sinusitis].
Methods of light, electron microscopy and immunohistochemistry were used to study the samples of maxillary sinus (MS) mucous membrane (MM) under normal conditions and in odontogenic sinusitis. To study the normal structure, the samples were obtained at autopsy from 26 human corpses 12-24 hours after death. Electron microscopic and immunohistochemical study was performed on biopsies of grossly morphologically unchanged MS MM, obtained during the operations for retention cysts in 6 patients. MS MM in perforative sinusitis was studied using the biopsies obtained from 43 patients. The material is broken into 4 groups depending on perforative sinusitis duration. Under normal conditions, MS MM is lined with a pseudostratified columnar ciliated epithelium. Degenerative changes of ciliated epithelial cells were already detected at short time intervals after MS perforations and become apparent due to reduction of specific volume of mitochondria and, rough endoplasmic reticulum, and increase of nuclear-cytoplasmic ratio. In the globlet cells, the reduction of nuclear-cytoplasmic ratio was associated with the disturbance of the secretory product release. At time intervals exceeding 3 months, epithelium underwent metaplasia into simple cuboidal and stratified squamous keratinized, while in MS MM lamina propria, cellular infiltration was increased. CD4+ cell content in sinus MM gradually increased, while at late periods after perforation occurrence it decreased. Low CD4+ cell count within the epithelium and the absence of muromidase on the surface of MS MM was detected. With the increase of the time interval since MS perforation, the number of CD8+ and CD20+ cells in MS MM was found to increase.


Label-free quantitative proteomic analysis of abscisic acid effect in early-stage soybean under flooding.
Flooding is a serious problem for soybean cultivation because it markedly reduces growth. To investigate the role of phytohormones in soybean under flooding stress, gel-free proteomic technique was used. When 2-day-old soybeans were flooded, the content of abscisic acid (ABA) did not decrease in the root, though its content decreased in untreated plant. When ABA was added during flooding treatment, survival ratio was improved compared with that of soybeans flooded without ABA. When 2-day-old soybeans were flooded with ABA, the abundance of proteins related to cell organization, vesicle transport and glycolysis decreased compared with those in root of soybeans flooded without ABA. Furthermore, the nuclear proteins were analyzed to identify the transcriptional regulation. The abundance of 34 nuclear proteins such as histone deacetylase and U2 small nuclear ribonucleoprotein increased by ABA supplementation under flooding; however, 35 nuclear proteins such as importin alpha, chromatin remodeling factor, zinc finger protein, transducin, and cell division 5 protein decreased. Of them, the mRNA expression levels of cell division cycle 5 protein, C2H2 zinc finger protein SERRATE, CCCH type zinc finger family protein, and transducin were significantly down-regulated under the ABA treatment. These results suggest that ABA might be involved in the enhancement of flooding tolerance of soybean through the control of energy conservation via glycolytic system and the regulation on zinc finger proteins, cell division cycle 5 protein and transducin.


The impact of routine cryptococcal antigen screening on survival among HIV-infected individuals with advanced immunosuppression in Kenya.
To test the hypothesis that a screening and treatment intervention for early cryptococcal infection would improve survival among HIV-infected individuals with low CD4 cell counts. Newly enrolled patients at Family AIDS Care and Education Services (FACES) in Kenya with CD4 ≤ 100 cells/μl were tested for serum cryptococcal antigen (sCrAg). Individuals with sCrAg titre ≥ 1:2 were treated with high-dose fluconazole. Cox proportional hazard models of Kaplan-Meier curves were used to compare survival among individuals with CD4 ≤ 100 cells/μl in the intervention and historical control groups. The median age was 34 years [IQR: 29,41], 54% were female, and median CD4 was 43 cells/μl [IQR: 18,71]. Follow-up time was 1224 person-years. In the intervention group, 66% (514/782) were tested for sCrAg; of whom, 11% (59/514) were sCrAg positive. Mortality was 25% (196/782) in the intervention group and 25% (191/771) in the control group. There was no significant difference between the intervention and control group in overall survival [hazard ratio (HR): 1.1 (95%CI:0.9,1.3)] or three-month survival [HR: 1.0 (95%CI:0.8,1.3)]. Within the intervention group, sCrAg-positive individuals had significantly lower survival rates than sCrAg-negative individuals [HR:1.8 (95%CI: 1.0, 3.0)]. A screening and treatment intervention to identify sCrAg-positive individuals and treat them with high-dose fluconazole did not significantly improve overall survival among HIV-infected individuals with CD4 counts ≤ 100 cells/μl compared to a historical control, perhaps due to intervention uptake rates or poor efficacy of high-dose oral fluconazole.


Phenotypic evaluation of the effect of anaerobiosis on some virulence attributes of Candida albicans.
The current assumption that Candida albicans is a facultatively anaerobic organism has been widely accepted since its recovery from anoxic sites became common. However, the link between anaerobiosis and virulence remains uncertain. This study investigated the differential cell-surface hydrophobicity (CSH) using a hydrocarbon/water partition technique and analysed the differential secretion rates of secretory aspartyl proteases (Saps), esterase, chondroitinase and haemolysins of C. albicans strains recovered from periodontal pockets and non-periodontium-related intra-oral sites. For the enzymic tests, all strains from both sets were grown under aerobic and anaerobic conditions and the harvested cells were inoculated onto suitable normal or pre-reduced culture media in the presence or absence of molecular oxygen, respectively. The results showed that no variations were perceptible for CSH and chondroitinase (P>0.05). The secretion rates of esterase and haemolysins strongly decreased in an anoxic environment (P<0.0001). However, a consistent increment (P<0.0001) in Sap secretion was detected when cultures were grown under anaerobic conditions. Based on these results, it is suggested that the oxygen concentration in the atmosphere surrounding cells exerts a variable influence on the virulence attributes of C. albicans.


Generation and functional significance of CXC chemokines for neutrophil-induced liver injury during endotoxemia.
The hypothesis that the neutrophil chemoattractant CXC chemokines KC and macrophage inflammatory protein-2 (MIP-2) are involved in neutrophil transmigration and liver injury was tested in C3Heb/FeJ mice treated with galactosamine (Gal, 700 mg/kg), endotoxin (ET, 100 microg/kg), or Gal + ET (Gal/ET). Hepatic KC and MIP-2 mRNA levels and plasma CXC chemokine concentrations were dramatically increased 1.5 h after Gal/ET or ET alone and gradually declined up to 7 h. Murine recombinant cytokines (TNF-alpha, IL-1 alpha, and IL-1 beta), but not Gal/ET, induced CXC chemokine formation in the ET-resistant C3H/HeJ strain. To assess the functional importance of KC and MIP-2, C3Heb/FeJ mice were treated with Gal/ET and control IgG or a combination of anti-KC and anti-MIP-2 antibodies. Anti-CXC chemokine antibodies did not attenuate hepatocellular apoptosis, sinusoidal neutrophil sequestration and extravasation, or liver injury at 7 h. Furthermore, there was no difference in liver injury between BALB/cJ wild-type and CXC receptor-2 gene knockout (CXCR2-/-) mice treated with Gal/ET. The higher neutrophil count in livers of CXCR2-/- than in wild-type mice after Gal/ET was caused by the elevated number of neutrophils located in sinusoids of untreated CXCR2-/- animals. The pancaspase inhibitor Z-Val-Ala-Asp-fluoromethylketone eliminated Gal/ET-induced apoptosis and neutrophil extravasation and injury but not CXC chemokine formation. Thus Gal/ET induced massive, cytokine-dependent CXC chemokine formation in the liver. However, neutrophil extravasation and injury occurred in response to apoptotic cell injury at 6-7 h and was independent of CXC chemokine formation.


[Centrioles and microtubules in interphase cells exposed to colcemid. The concentration- and time-dependent effect of the action of the poison].
Under the action of colcemid on SPEV cells the network of cytoplasmic microtubules disappears within less than 1 hour; microtubules attached to pericentriolar satellites are retained for 4 hours. The disassembly time of these microtubules does not depend on colcemide concentration. It is therefore assumed that most of the microtubules are not attached to the centrioles, but have two free ends, thus confirming a hypothesis that they are conveyer-assembled. With colcemid concentration equal to 0.5 mcg/ml, the following dynamics of events is observed for the cell centre: after the microtubules attached to the satellites had disappeared, clusters of electron dense material appear around the centrioles (6 hour incubation), then short microtubules occur among clusters (8 hour incubation) to be subsequently retained (up to 40 hour incubation).


Ethylene and phosphorus availability have interacting yet distinct effects on root hair development.
The hypothesis that ethylene participates in the regulation of root hair development by phosphorus availability in Arabidopsis thaliana was tested by chemically manipulating ethylene synthesis and response and with ethylene-insensitive mutants. Low phosphorus-induced root hair development could be mimicked by adding the ethylene precursor, 1-aminocyclopropane-1-carboxylate (ACC), to high phosphorus media, and inhibited by adding ethylene inhibitors to low phosphorus media. Ethylene-insensitive mutants showed a reduced response to low phosphorus, indicating ethylene involvement in root hair responses to phosphorus deficiency. To dissect the nature of this involvement, the morphological and anatomical changes associated with increased root hair density were investigated. Growth in low phosphorus resulted in smaller, more numerous cortical cells, resulting in a larger number of root hair-bearing epidermal cell files. Cortical cell number was not affected by ethylene inhibitors, ACC, or mutations reducing ethylene sensitivity in roots grown with low phosphorus, indicating that ethylene does not participate in this response. The exception was the eir1 mutation, which strongly reduced this change in radial anatomy, supporting a role for polar auxin transport in this process. Trichoblast cell length was reduced by low phosphorus availability in all genotypes, but even more so for ein2-1 and ein4. The proportion of epidermal cells forming hairs and root hair length were reduced in ethylene-insensitive mutants, especially in the presence of low phosphorus. These results demonstrate multiple effects of low phosphorus from the earliest stages of root hair development, and cross-talk between ethylene and phosphorus in the control of a subset of the low phosphorus effects, concentrating on those later in development.


A case of steatocystoma multiplex limited to the scalp.
We report here on a 63-year-old woman who had several small, yellowish papules on the scalp for the previous 2 years. There was no family history of similar lesions. Yellowish, creamy material was expressed from a papule during punch biopsy. Histologic examination from the lesion revealed the typical features of steatocystoma multiplex. We report here on this rare variant of steatocystoma multiplex that was limited to the scalp.


Pattern and Prognostic Impact of Regional Wall Motion Abnormalities in 255 697 Men and 236 641 Women Investigated with Echocardiography.
Background Regional wall motion abnormalities (WMAs) after myocardial infarction are associated with adverse remodeling and increased mortality in the short to medium term. Their long-term prognostic impact is less well understood. Methods and Results Via the National Echo Database of Australia (2000-2019), we identified normal wall motion versus WMA for each left ventricular wall among 492 338 individuals aged 61.9±17.9 years. The wall motion score index was also calculated. We then examined actual 1- and 5-year mortality, plus adjusted risk of long-term mortality according to WMA status. Overall, 39 346/255 697 men (15.4%) and 17 834/236 641 women (7.5%) had a WMA. The likelihood of a WMA was associated with increasing age and greater systolic/diastolic dysfunction. A defect in the inferior versus anterior wall was the most and least common WMA in men (8.0% and 2.5%) and women (3.3% and 1.1%), respectively. Any WMA increased 5-year mortality from 17.5% to 29.7% in men and from 14.9% to 30.8% in women. Known myocardial infarction (hazard ratio [HR], 0.86 [95% CI, 0.80-0.93]) or revascularization (HR, 0.87 [95% CI, 0.82-0.92]) was independently associated with a better prognosis, whereas men (1.22-fold increase) and those with greater systolic/diastolic dysfunction had a worse prognosis. Among those with any WMA, apical (HR, 1.08 [95% CI, 1.02-1.13]) or inferior (HR, 1.09 [95% CI, 1.04-1.15]) akinesis, dyskinesis or aneurysm, or a wall motion score index >3.0 conveyed the worst prognosis. Conclusions In a large real-world clinical cohort, twice as many men as women have a WMA, with inferior WMA the most common. Any WMA confers a poor prognosis, especially inferoapical akinesis/dyskinesis/aneurysm.


Synthesis and in vitro antiproliferative and antibacterial activity of new thiazolidine-2,4-dione derivatives.
In our present research, we synthesised new thiazolidine-2,4-diones (12-28). All the newly synthesised compounds were evaluated for antiproliferative and antibacterial activity. Antiproliferative evaluation was carried out using normal human skin fibroblasts and tumour cell lines: A549, HepG2, and MCF-7. The IC50 values were determined for tested compounds revealing antiproliferative activity. Moreover, safety index (SI) was calculated. Among all tested derivatives, the compound 18 revealed the highest antiproliferative activity against human lung, breast, and liver cancer cells. More importantly, the derivative 18 showed meaningfully lower IC50 values when compared to the reference substance, irinotecan, and relatively high SI values. Moreover, newly synthesised compounds were screened for the bacteria growth inhibition in vitro. According to our screening results, most active compound was the derivative 18 against Gram-positive bacteria. Therefore, it may be implied that the novel compound 18 appears to be a very promising agent for anticancer treatment.


The effect of CD14 C159T polymorphism on in vitro IgE synthesis and cytokine production by PBMC from children with asthma.
Even though the genotype at the promoter region of the CD14 molecule is known to affect the atopic phenotypes, the cellular and molecular basis of this association is largely unknown. To investigate the effect of lipopolysaccharide (LPS) on IgE production and cytokine profile by peripheral blood mononuclear cells (PBMC) obtained from asthmatic children with the TT and the CC genotypes at position -159 of the CD14 gene. Peripheral blood mononuclear cells from asthmatic children with alternative genotypes at CD14 C159T locus were stimulated with 2 and 200 ng/ml LPS in vitro. The IgE, IgG and, IgM response was determined by ELISA and Ig έ-germline, IgG, and IgM transcription by real-time PCR. A cluster of cytokines was measured by cytometric bead array. Asthmatic children with the TT genotype but not those with the CC genotype responded with increased IgE synthesis and germline transcription to LPS stimulation. There were no genotype-related differences in IgG and IgM. TT but not the CC genotype was associated with significantly increased interleukin (IL)-4/IL-12 and IL-4/interferon-gamma (IFN-γ) ratios in the culture supernatant. There were no genotype-related differences in IL-1β, IL-7, IL-10, IL-13, IL-17A, granulocyte colony stimulating factor, granulocyte macrophage colony stimulating factor, monocyte chemotactic protein, and tumor necrosis factor alpha. Peripheral blood mononuclear cells from asthmatic children with the TT genotype at position -159 of the CD14 gene make more IgE than those with the CC genotype following LPS stimulation because of increased germline transcription and have an augmented Th2 cytokine profile.


Critical prosurvival roles for C/EBP beta and insulin-like growth factor I in macrophage tumor cells.
One of the hallmarks of leukemic cells is their ability to proliferate and survive in the absence of exogenous growth factors (GFs). However, the molecular mechanisms used by myeloid tumor cells to escape apoptosis are not fully understood. Here we report that Myc/Raf-transformed macrophages require the transcription factor C/EBP beta to prevent cell death. In contrast to wild-type cells, C/EBP beta(-/-) macrophages were completely dependent on macrophage colony-stimulating factor or granulocyte-macrophage colony-stimulating factor for survival and displayed impaired tumorigenicity in vivo. Microarray analysis revealed that C/EBP beta-deficient cells expressed significantly reduced levels of the prosurvival factor insulin-like growth factor I (IGF-I). Overexpression of C/EBP beta stimulated transcription from the IGF-I promoter, indicating that IGF-I is a direct transcriptional target of C/EBP beta. Serological neutralization of IGF-I in C/EBP beta(+/+) tumor cell cultures induced apoptosis, showing that IGF-I functions as an autocrine survival factor in these cells. Macrophage tumor cells derived from IGF-I(-/-) mice were GF dependent, similar to C/EBP beta-deficient cells. Forced expression of either C/EBP beta or IGF-I in C/EBP beta(-/-) bone marrow cells restored Myc/Raf-induced transformation and permitted neoplastic growth without exogenous GFs. Thus, our findings demonstrate that C/EBP beta is essential for oncogenic transformation of macrophages and functions at least in part by regulating expression of the survival factor IGF-I.


A demonstration of attentional bias, using a novel dual task paradigm, towards clinically salient material in recovering alcohol abuse patients?
This study examined whether alcohol abuse patients are characterized either by enhanced schematic processing of alcohol related cues or by an attentional bias towards the processing of alcohol cues. Abstinent alcohol abusers (N = 25) and non-clinical control participants (N = 24) performed a dual task paradigm in which they had to make an odd/even decision to a centrally presented number while performing a peripherally presented lexical decision task. Stimuli on the lexical decision task comprised alcohol words, neutral words and non-words. In addition, participants completed an incidental recall task for the words presented in the lexical decision task. It was found that, in the presence of alcohol related words, the performance of patients on the odd/even decision task was poorer than in the presence of other stimului. In addition, patients displayed slower lexical decision times for alcohol related words. Both groups displayed better recall for alcohol words than for other stimuli. These results are interpreted as supporting neither model of drug cravings. Rather, it is proposed that, in the presence of alcohol stimuli, alcohol abuse patients display a breakdown in the ability to focus attention.


A study of statistics anxiety levels of graduate dental hygiene students.
In light of increased emphasis on evidence-based practice in the profession of dental hygiene, it is important that today's dental hygienist comprehend statistical measures to fully understand research articles, and thereby apply scientific evidence to practice. Therefore, the purpose of this study was to investigate statistics anxiety among graduate dental hygiene students in the U.S. A web-based self-report, anonymous survey was emailed to directors of 17 MSDH programs in the U.S. with a request to distribute to graduate students. The survey collected data on statistics anxiety, sociodemographic characteristics and evidence-based practice. Statistic anxiety was assessed using the Statistical Anxiety Rating Scale. Study significance level was α=0.05. Only 8 of the 17 invited programs participated in the study. Statistical Anxiety Rating Scale data revealed graduate dental hygiene students experience low to moderate levels of statistics anxiety. Specifically, the level of anxiety on the Interpretation Anxiety factor indicated this population could struggle with making sense of scientific research. A decisive majority (92%) of students indicated statistics is essential for evidence-based practice and should be a required course for all dental hygienists. This study served to identify statistics anxiety in a previously unexplored population. The findings should be useful in both theory building and in practical applications. Furthermore, the results can be used to direct future research.


Paired-like homeobox gene (PHOX2B) nonpolyalanine repeat expansion mutations (NPARMs): genotype-phenotype correlation in congenital central hypoventilation syndrome (CCHS).
CCHS is an extremely rare congenital disorder requiring artificial ventilation as life support. Typically caused by heterozygous polyalanine repeat expansion mutations (PARMs) in the PHOX2B gene, identification of a relationship between PARM length and phenotype severity has enabled anticipatory management. However, for patients with non-PARMs in PHOX2B (NPARMs, ~10% of CCHS patients), a genotype-phenotype correlation has not been established. This comprehensive report of PHOX2B NPARMs and associated phenotypes, aims at elucidating potential genotype-phenotype correlations that will guide anticipatory management. An international collaboration (clinical, commercial, and research laboratories) was established to collect/share information on novel and previously published PHOX2B NPARM cases. Variants were categorized by type and gene location. Categorical data were analyzed with chi-square and Fisher's exact test; further pairwise comparisons were made on significant results. Three hundred two individuals with PHOX2B NPARMs were identified, including 139 previously unreported cases. Findings demonstrate significant associations between key phenotypic manifestations of CCHS and variant type, location, and predicted effect on protein function. This study presents the largest cohort of PHOX2B NPARMs and associated phenotype data to date, enabling genotype-phenotype studies that will advance personalized, anticipatory management and help elucidate pathological mechanisms. Further characterization of PHOX2B NPARMs demands longitudinal clinical follow-up through international registries.


The essential genomic landscape of the commensal Bifidobacterium breve UCC2003.
Bifidobacteria are common gut commensals with purported health-promoting effects. This has encouraged scientific research into bifidobacteria, though recalcitrance to genetic manipulation and scarcity of molecular tools has hampered our knowledge on the precise molecular determinants of their health-promoting attributes and gut adaptation. To overcome this problem and facilitate functional genomic analyses in bifidobacteria, we created a large Tn5 transposon mutant library of the commensal Bifidobacterium breve UCC2003 that was further characterized by means of a Transposon Directed Insertion Sequencing (TraDIS) approach. Statistical analysis of transposon insertion distribution revealed a set of 453 genes that are essential for or markedly contribute to growth of this strain under laboratory conditions. These essential genes encode functions involved in the so-called bifid-shunt, most enzymes related to nucleotide biosynthesis and a range of housekeeping functions. Comparison to the Bifidobacterium and B. breve core genomes highlights a high degree of conservation of essential genes at the species and genus level, while comparison to essential gene datasets from other gut bacteria identified essential genes that appear specific to bifidobacteria. This work establishes a useful molecular tool for scientific discovery of bifidobacteria and identifies targets for further studies aimed at characterizing essential functions not previously examined in bifidobacteria.


Lemierre syndrome: from pharyngitis to fulminant sepsis.
We report a case of a previously healthy 33-year-old male who presented to his primary care physician with nausea, vomiting, diarrhoea and fever. One week prior to presentation the patient reported a history of sore throat which he presumed to be a viral infection and sought no medical attention. Upon hospital presentation, the patient was admitted and rapidly progressed to sepsis and respiratory failure. Goal directed therapy was initiated and the patient was intubated. Further clinical work up included blood cultures revealing Fusobacterium varium bacteraemia, and CT and ultrasound imaging demonstrated thrombosis of the internal jugular vein and septic pulmonary emboli. A diagnosis of Lemierre syndrome was made, and antibiotics as well as anticoagulation therapy were initiated. The patient's clinical condition improved with treatment, and he was discharged home on hospital day 12 with completion of an uneventful 4-week course of outpatient antibiotic and anticoagulation therapy.


The attribution of somatization in schizophrenia patients: a naturalistic follow-up study.
Knowledge is limited concerning somatic symptoms that cannot be accounted for by detectable somatic illness among schizophrenia patients. This study aimed to explore the prevalence, correlates, and predictors of somatization among schizophrenia patients. Initial data on all consecutively admitted adult patients with DSM-IV schizophrenia, schizoaffective disorder, major depressive disorder, or bipolar disorder were collected between August 1998 and August 2000. Standardized measures of psychopathology, somatization, emotional distress, adverse effects, insight, and stress process-related (psychosocial) variables were administered to 237 schizophrenia patients at admission and at least 12 months thereafter (N = 148). Partial correlation and multiple regression analyses were performed. The frequency of somatization, defined as the presentation of 5 or more medically unexplained somatic symptoms (Somatic Symptom Index-5), among the inpatient population (27%) did not change significantly after at least 12 months (30%; p =.61). Regression analysis showed that somatization scores were best predicted by the combination of scores for emotional distress attributed to psychopathology and side effects, expressed emotion, and insight. This combined model explains at least 40% of the variance in somatization scores. Self-esteem and social support showed negative association with somatization scores. Somatization scores were not associated with gender, age, education, age at onset, observed severity of psychopathology, subtype and duration of illness, number of admissions and treatment settings, or type and dose of antipsychotic agents. Somatization is a prevalent problem among schizophrenia patients and is associated with emotional distress attributed to psychopathology, side effects of antipsychotic agents, and family members' attitudes toward schizophrenia patients. This study suggests that insight, self-esteem, and social support may protect against somatization in schizophrenia patients.


Prevalence of Early Childhood Caries (ECC) in a paediatric italian population: An epidemiological study.
The present work aims to evaluate, through an epidemiological study, the prevalence of Early Childhood Caries in an Italian paediatric population, aged up to 71 months, evaluating some determinants that will be illustrated in this study in order to adopt preventive care and intervene as early as possible to limit the development of this disease. The sample consisted of 76 children aged up to 71 months who were examined at the PAediatric Dentistry Clinic of the University of l'Aquila. Parents were informed of the research that was taking place. All read the information sheet explaining the study and signed the informed consent where it was emphasised that the data collected would be treated and stored in a strictly confidential manner, pursuant to the Legislative Decree 196/2003 in force in Italy since June 30, 2003, regarding the protection of individuals with regard to the processing of personal data. At the end of this study, a standardised questionnaire administered. The questionnaire, divided into two parts, made it possible to acquire information regarding the characteristics of the population. First part: General data of the child and the medical history of the mother and father; Second part: The age and gender of the child, the type and duration of breastfeeding, the use of sugary pacifiers, oral hygiene and frequency of consumption of sugars drinks and food, identifying the starting age. Evaluating the dmft of each child, 40.79% of our sample population is affected by ECC; 57.89% of the children used the pacifier and 13,16% of them have or have used it with honey with a p value of 0.001, therefore this parameter was statistically significant, together with the intake of candies, chocolates and sugar drinks. The data was collected for a period of about 12 months, consequently a descriptive statistical analysis was conducted in which the characteristics of the population under consideration were outlined. Discrete and nominal variables have been described with frequencies and percentages. The percentage differences were evaluated using the chi-square test and the Fisher test. Quantitative variables were expressed in terms of mean and standard deviation and significance was assessed by the Mann-Whitney test. The level of significance was established as p <0.05. A Logistic Regression was conducted to evaluate the risk factors associated with a dmft>0 setting, dmft=0/dmft>0 as the dependent variable and the variables investigated as explanatory variables. The evaluation of the association was reported as the Odds Ratio with the 95% confidence interval. The statistical analysis was carried out using the Stata 12/1C statistical package. ECC is a multifactorial disease in which eating habits play a very important role. A diet that from childhood is characterised by a high consumption of sugar-rich food and drinks is highly associated with the incidence of ECC in later years.


Receptor-mediated MAP kinase activation in plant defense.
Plants mount a complex array of defense reactions in response to attack by pathogens. Initiation of these events depends on perception and signal transduction of elicitors, which are plant-derived or pathogen-derived signals, that give rise to transcriptional activation of defense-related genes as well as to changes in activities of enzymes involved in cell wall reinforcement and oxygen radical formation. An oligopeptide, identified within a 42 kDa glycoprotein elicitor from Phythophthora sojae, activates in parsley cells typical plant defense reactions, enabling researchers to study plant-pathogen interaction at the single cell level. The oligopeptide elicitor was found to be necessary and sufficient to stimulate a complex defense response in parsley cells, comprising H+/Ca2+ influxes, K+/Cl- effluxes, activation of a mitogen-activated protein (MAP) kinase, an oxidative burst, defense-related gene activation, and phytoalexin formation.


Isolation of the glycoprotein of vesicular stomatitis virus and its binding to cell surfaces.
The glycoprotein (G) of vesicular stomatitis virus (VSV) was radiolabelled, extracted and purified so that its potential interaction with host cell surfaces could be studied. When BHK-21 cells were incubated with the radiolabelled virus glycoprotein, the virus component rapidly attached to the cell surface. The attachment was shown to be temperature-dependent adn saturated at approx. 3 X 10(5) molecules/cell. The omission of Mg2+ or Ca2+ from the incubation medium had little effect on the glycoprotein binding. Treating the isolated G protein and intact virions with neuraminidase did not significantly decrease their binding to BHK-21 cells. Pre-incubating cells with trypsin did not decrease the attachment of VSV virions nor the binding of purified G protein. Treating cells with phospholipase A or phospholipase C suggested that the binding of the glycoprotein and the intact virion might have been dissimilar. Unlabelled glycoprotein competitively inhibited binding of the labelled molecules although the presence of intact virions did not inhibit attachment of the G protein. Likewise, saturating amounts of the glycoprotein did not decrease binding of VSV to BHK-21 cells. These observations suggested that either the isolated glycoprotein bound to cell surface components that were distinct from the virion receptor or that the manner of the purified glycoprotein attachment differed from the G protein still associated with the intact virion. Chemical crosslinking and diagonal two-dimensional gel electrophoresis were used to identify and to compare the cell surface components responsible for glycoprotein and virion attachment.


A new method for scintigraphic quantification of deposition and clearance in anatomical regions of the human nose.
To develop methods for absolute quantification of the deposition of 99mTc-labeled aerosols and powders in well-defined anatomical regions of the nose, and to enable accurate comparisons of different nasal administration techniques in the same individual. The volunteer was seated and positioned relative to the scintillation camera field of view by means of a fixation frame. After nasal administration, a dynamic series of images was acquired for 32 min with a lateral direction of view. The images were corrected for photon attenuation by the use of a lateral transmission image acquired before the delivery of aerosols or powders. Marker images, obtained with a line source fixed to a balloon and kept for a short while against the palate as well as with a point source held on anatomical landmarks, were used to co-register the scintigraphic images to sagital sections through a three-dimensional magnetic resonance (MR)-image series. The MR set was used to define the inner nose contour and the nasal regions used for quantification. Attenuation correction factors ranged from 1.1 to 1.7 in different parts of the nasal cavity. Alignment of the markers on the teeth and palate with the sagital MR images could be reproduced with accuracies of 1.2 and 1.7 mm, respectively. The new method provides reliable quantification of the deposition in anatomic regions that can be defined in MR images. Accurate co-registration and quantification are essential when comparing distribution and clearance patterns for different administration techniques.