Clinical and pathological feature of bone marrow granulomas: A modern Australian series.
Marrow granulomas have been commonly associated with mycobacterial infections, prompting extensive investigations in some cases where they are found, but in the setting of changing infectious epidemiology, there is a lack of recent data to guide the search for probable causes. A retrospective case series examining the clinical and pathological findings in all cases where marrow granulomas were reported over a 10-year period at The Canberra Hospital, Australia. Of the 6062 marrow biopsies performed, 73 (1.2%) from 66 individuals had confirmed granulomas. Some patients had multiple potential causes found. B-cell non-Hodgkin Lymphoma, sarcoidosis and autoimmune disease were the most frequently observed causes. Infections were less common in this cohort than in the previous series, with no tuberculosis seen. The presence of granulomas in the marrow did not signify the presence of active malignancy in the marrow. There were no associations with any specific morphological characteristics of the granulomas and the presumed causes. Marrow granulomas are seen in a variety conditions. Neither their presence nor their morphological features are a guide to further investigations, which should be determined by the clinical presentation as appropriate.


Analgesia induced by cold-water stress: attenuation following hypophysectomy.
In addition to the well-known activation of the pituitary-adrenal axis, acute exposure to severe stressors includes a temporary analgesia in rats. Thus, the present study investigates whether the pituitary was involved in the mediation of analgesia induced by severe cold-water swim (CWS) stress. Flinch-jump thresholds were measured 30 min following 3.5-min swims in water temperatures ranging from 2-35 degrees C. Compared with untreated normal rats, hypophysectomized rats, receiving corticosterone and thyroxin, displayed significantly less CWS-induced analgesia, while similarly-supplemented normal rats exhibited significantly more CWS-induced analgesia. In a second experiment, operant liminal escape pain thresholds were determined following acute and chronic CWS. Whereas normal rats exhibited profound analgesia following the initial swims, the hypophysectomized rats never displayed any CWS-induced operant escape shifts. Stress-induced alterations in general activity levels and/or thermoregulation were shown to be unrelated to the diminished effectiveness of CNS to produce analgesia in hypophysectomized rats. These data imply that the pituitary is involved in the mediation of CWS-induced analgesia.


Association between second- and third-trimester maternal lipid profiles and adverse perinatal outcomes among women with GDM and non-GDM: a retrospective cohort study.
Lipid metabolism disorder during pregnancy has been reported in women with gestational diabetes mellitus (GDM). However, controversy remains regarding the relationship between maternal changes in lipid profiles and perinatal outcomes. This study investigated the association between maternal lipid levels and adverse perinatal outcomes in women with GDM and non-GDM. In total, 1632 pregnant women with GDM and 9067 women with non-GDM who delivered between 2011-2021 were enrolled in this study. Serum samples were assayed for fasting total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) levels during the second and third trimesters of pregnancy. Adjusted odds ratios (AOR) and 95% confidence intervals (95% CI) were calculated via multivariable logistic regression analysis to determine the association of lipid levels with perinatal outcomes. The serum TC, TG, LDL, and HDL levels in the third trimester were significantly higher than those in the second trimester (p < 0.001). Women with GDM had significantly higher levels of TC and TG in the second and third trimesters than those with non-GDM in the same trimesters, while HDL levels decreased in women with GDM (all p < 0.001). After adjusting for confounding factors by multivariate logistic regression, every mmol/L elevation in TG levels of women with GDM in second and third trimesters was associated with a higher risk of caesarean section (AOR = 1.241, 95% CI: 1.103-1.396, p < 0.001; AOR = 1.716, 95% CI: 1.556-1.921, p < 0.001), large for gestational age infants (LGA) (AOR = 1.419, 95% CI: 1.173-2.453, p = 0.001; AOR = 2.011, 95% CI: 1.673-2.735, p < 0.001), macrosomia (AOR = 1.220, 95% CI: 1.133-1.643, p = 0.005; AOR = 1.891, 95% CI: 1.322-2.519, p < 0.001), and neonatal unit admission (NUD; AOR = 1.781, 95% CI: 1.267-2.143, p < 0.001; AOR = 2.052, 95% CI: 1.811-2.432, p < 0.001) cesarean delivery (AOR = 1.423, 95% CI: 1.215-1.679, p < 0.001; AOR = 1.834, 95% CI: 1.453-2.019, p < 0.001), LGA (AOR = 1.593, 95% CI: 1.235-2.518, p = 0.004; AOR = 2.326, 95% CI: 1.728-2.914, p < 0.001), macrosomia (AOR = 1.346, 95% CI: 1.209-1.735, p = 0.006; AOR = 2.032, 95% CI: 1.503-2.627, p < 0.001), and neonatal unit admission (NUD) (AOR = 1.936, 95% CI: 1.453-2.546, p < 0.001; AOR = 1.993, 95% CI: 1.724-2.517, p < 0.001), which were higher than the relative risk of these perinatal outcomes in women with non-GDM. Additionally, every mmol/L increase in second and third-trimester HDL levels of women with GDM was associated with decreased risk of LGA(AOR = 0.421, 95% CI: 0.353-0.712, p = 0.007; AOR = 0.525, 95% CI: 0.319-0.832, p = 0.017) and NUD (AOR = 0.532, 95% CI: 0.327-0.773, p = 0.011; AOR = 0.319, 95% CI: 0.193-0.508, p < 0.001), and the risk reduction was not strong than that of women with GDM. Among women with GDM, high maternal TG in the second and third trimesters was independently associated with an increased risk of cesarean section, LGA, macrosomia, and NUD. High maternal HDL during the second and third trimesters was significantly associated with decreased risk of LGA and NUD. These associations were stronger than those in women with non-GDM, suggesting the importance of monitoring second and third-trimester lipid profiles in improving clinical outcomes, especially in GDM pregnancies.


[Melitococcal hepatic encephalopathy].
A 5 year old child with brucellosis is described who developed severe liver disease with neuropsychiatric signs and electroencephalographic changes.


BRK phosphorylates SMAD4 for proteasomal degradation and inhibits tumor suppressor FRK to control SNAIL, SLUG, and metastatic potential.
The tumor-suppressing function of SMAD4 is frequently subverted during mammary tumorigenesis, leading to cancer growth, invasion, and metastasis. A long-standing concept is that SMAD4 is not regulated by phosphorylation but ubiquitination. Our search for signaling pathways regulated by breast tumor kinase (BRK), a nonreceptor protein tyrosine kinase that is up-regulated in ~80% of invasive ductal breast tumors, led us to find that BRK competitively binds and phosphorylates SMAD4 and regulates transforming growth factor-β/SMAD4 signaling pathway. A constitutively active BRK (BRK-Y447F) phosphorylates SMAD4, resulting in its recognition by the ubiquitin-proteasome system, which accelerates SMAD4 degradation. Activated BRK-mediated degradation of SMAD4 is associated with the repression of tumor suppressor gene FRK and increased expression of mesenchymal markers, SNAIL, and SLUG. Thus, our data suggest that combination therapies targeting activated BRK signaling may have synergized the benefits in the treatment of SMAD4 repressed cancers.


Catecholaminergic Polymorphic Ventricular Tachycardia in Pregnancy.
Catecholaminergic polymorphic ventricular tachycardia is a genetic disorder in which ventricular tachycardia occurs in the absence of structural heart disease or a prolonged QT interval. If untreated, there is a high incidence of sudden cardiac death. Management of this cardiac condition during pregnancy merits a multidisciplinary approach. A nulliparous woman with catecholaminergic polymorphic ventricular tachycardia presented at 15 weeks of gestation. Her care involved a multidisciplinary team including cardiology, maternal-fetal medicine, obstetric nursing, cardiac nursing, and anesthesia. A simulation scenario was designed to prepare for cardiac events during labor. A term intrapartum cesarean delivery was performed for fetal indications. A multidisciplinary approach to the antepartum, intrapartum, and postpartum care of women with catecholaminergic polymorphic ventricular tachycardia is critical to a team-based successful pregnancy outcome.


Relationship between prostatic urethral angle and urinary flow rate: its implication in benign prostatic hyperplasia pathogenesis.
The prostatic urethra is a bent tube, and the clinical significance of the prostatic urethral angle (PUA) is poorly understood. We investigated the effect of the PUA on the urinary flow rate in men without benign prostatic obstruction. In a theoretical model, the urinary flow rate at the end of the prostatic urethra was formulated according to some principles of fluid dynamics, including the Navier-Storkes equation, Bernoulli's equation, and the loss of coefficient at bends. The change in the urinary flow rate according to the PUA was also calculated through mathematical simulations. In addition, the relationship between the PUA and the peak flow rate was evaluated in 65 healthy men without benign prostatic obstruction (age 50 to 59 years, prostate volume less than 30 cm(3), peak flow rate greater than 15 mL/s). The prostate volume and PUA were determined by transrectal ultrasonography. Using the fluid dynamic model, the urinary flow rate increased in proportion to the urethral diameter and was inversely associated with the PUA. The mathematical simulations showed that the urinary flow rate decreased by more than 27% as the PUA increased from 35 degrees to 90 degrees. In the clinical setting, the peak flow rate was significantly associated with the PUA by linear regression analysis (R = 0.34, beta = -0.342, P = 0.005), but it was not associated with the total prostate volume (R = 0.18, beta = -0.181, P = 0.150). The PUA was inversely associated with the urinary flow rate in this theoretical model and preliminary clinical study. This relationship could be an important factor in the pathogenesis of benign prostatic hyperplasia/lower urinary tract symptoms.


Local but not systemic capillary lactate is a reperfusion biomarker in experimental acute limb ischaemia.
Systemic capillary lactate, an end product of cellular anaerobic metabolism, has not established credibility in monitoring limb reperfusion. We assessed, in mice, whether local capillary lactate, arising from the reperfused limb, might be a relevant biomarker of reperfusion. Systemic and local capillary lactate were sampled in the non-ischaemic and in the ischaemic limb. Only local lactate concentrations significantly increased after 2 h of ischaemia and decreased after reperfusion. Local, but not systemic, capillary lactate appeared as a potential reperfusion biomarker in this experimental acute limb ischaemia model.


The effect of continued feeding of physiological amounts of lactose on the level of intestinal lactase and other disaccharidase enzyme activities in the rat.
Intestinal lactase activity in mammals is high at birth but begins to decline around weaning and reaches very low levels in adult life. The triggering mechanism for this decline is not clear. Because of the association of the decline with weaning, lack of lactose in the diet has been implicated. In 110 growing rats, the effect of continued supplementation of the diet after weaning with physiological amounts of either cows' milk or a 5% lactose solution on intestinal lactase and other disaccharidase enzyme activities was investigated. In both control and test animals, the specific lactase activity decreased from a peak value of 115 +/- 4 mumol min-1 g-1 protein before weaning to about 10% at maturity. There was no significant difference in the level or the pattern of decline between the groups. Sucrase, maltase and trehalase showed the normal maturational changes without being affected by the test diets. The finding suggests that diet, particularly the presence or absence of physiological amounts of lactose, has no appreciable effect on the age related spontaneous decline of intestinal lactase activity or on the pattern of development of the other disaccharidases.


Self-Forgiveness Moderates the Effects of Depression on Suicidality.
Not all depressive individuals are suicidal. An increasing body of studies has examined forgiveness, especially self-forgiveness, as a protective factor of suicide based on that suicide is often accompanied by negative self-perceptions. However, less has been studied on how different subtypes of forgiveness (i.e., forgiveness-of-self, forgiveness-of-others and forgiveness-of-situations) could alleviate the effects of depression on suicide. Hence, this study examined forgiveness as a moderator of depression and suicidality. 305 participants, consisted of 87 males and 218 females, were included in the study. The mean age was 41.05 (SD: 14.48; range: 19-80). Depression, anxiety, and forgiveness were measured through self-report questionnaires, and suicidal risk was measured through a structuralized interview. Moderations were examined through hierarchical regression analyses. Depression positively correlated with suicidality. of the hierarchical regression analysis indicated forgiveness as a moderator of depression on suicidality. Further analysis indicated only forgiveness-of-self as a significant moderator; the effects of forgiveness-of-others and forgiveness-of-situation were not significant. These findings suggest that forgiveness-of-self is essential in reducing of the effects of depression on suicidality. It is suggested that self-acceptance and the promotion of self-forgiveness should be considered as an important factor when developing suicide prevention strategies.


Cellular mitochondrial heterogeneity in cultured astrocytes as demonstrated by immunogold labeling of alpha-ketoglutarate dehydrogenase.
In brain cells, various metabolites and metabolic pathways, largely of mitochondrial origin, have been shown to be compartmentalized. Attention has therefore been focused on the possible existence of mitochondrial heterogeneity in the brain at the cellular level. To determine whether mitochondria in cultured cortical and cerebellar astrocytes are heterogeneous at the single cell level, immunogold electron microscopy and an antibody against the alpha-ketoglutarate dehydrogenase component of the alpha-ketoglutarate dehydrogenase complex, a marker enzyme for the tricarboxylic acid (TCA) cycle, were employed. The number of gold particles was counted in the mitochondria of 36 and 42 cells from cultured cerebellar and cortical astrocytes, respectively. A test for random distribution (Poisson distribution) of mitochondria according to the number of gold particles was subsequently performed for every one of the 36 and 42 cells as the ratio variance/mean (= index of dispersion). This should be approximately distributed as chi2/degrees of freedom (df) = n - 1, n = number of mitochondria), if the observations obeyed a Poisson distribution. For 26 of the 36 (cerebellar astrocytes) distributions and for 28 of the 42 (cortical astrocytes) distributions a random distribution had to be rejected. These findings therefore strongly indicate that alpha-ketoglutarate dehydrogenase is heterogeneously distributed in mitochondria within individual astrocytes originating either from cerebellum or cerebral cortex. In conclusion, this study underlines the probability that mitochondrial heterogeneity at the single cell level might be extended to involve other metabolic pathways and metabolites.


Inhibition of host protein synthesis in B95a cells infected with the HL strain of measles virus.
The shut-off of host protein synthesis in virus-infected cells is one of the important mechanisms for viral replication. In this report, we showed that the HL strain of measles virus (MeV-HL) as well as other field isolates, which were isolated from human blood lymphocytes using B95a cells, induce the shut-off in B95a cells. Since the Edmonston strain of MeV failed to induce the shut-off in B95a cells, the ability to induce the shut-off was considered to be dependent on virus strains. Although, the modification of eukaryotic translation initiation factors (eIF) including eIF4G, eIF4E, and 4E-BP1 was reported for shut-off by various viruses, the involvement of these eIFs was not observed in MeV-HL-infected B95a cells. Instead, the accumulation of phosphorylated eIF2alpha was found to coincide to the decrease of host protein synthesis, suggesting the involvement of phosphorylation of eIF2alpha in inhibition of translation as one of the mechanisms of the shut-off.


Past 50 years, present, and future of the Korean Nutrition Society.
Created in 1967, the Korean Nutrition Society (KNS) has been a vital contributor to the development of the health and nutrition of South Korea. With members from many different fields focusing on nutrition and health sciences, the impact of the KNS has not only been felt in South Korea, but also worldwide. The KNS has helped to establish and maintain dietary recommendations in addition to providing ways to make nutrition information readily available to members, as well as the public. The KNS has also established professional journals to advance scientific discoveries in nutrition and health. With 50 years of progress from the KNS, their contributions to nutrition and health will continue to have an impact throughout the world. To celebrate the 50th anniversary and growth of the KNS, an International Conference entitled "Integrative Nutrition for an Active Life" was held by the KNS in November 2017. The articles in this special issue of Nutrition Research are to commemorate the KNS efforts.


Abrogation of red blood cell G6PD enzyme activity through Heat treatment: development of survey material for the UK NEQAS G6PD scheme.
Participation in external quality assessment (EQA) is central to the maintenance of high-quality laboratory results in patient diagnosis and clinical trials. Laboratories in the TAF112582 DETECTIVE study (ClinicalTrials.gov identifier: NCT01376167) are enrolled in the United Kingdom National Quality Assessment Scheme (UK NEQAS) for glucose-6-phosphate dehydrogenase (G6PD) quantitative assay, which utilizes ovine (sheep) blood as a readily available source of apparently G6PD-deficient survey material. A substitute for sheep blood was sought because some non-UK sites in the study encountered participation difficulties due to the strict regulations on the import of sheep blood into their countries. G6PD activity in normal human donor blood was abrogated by the action of heat under controlled conditions. Residual G6PD activity in the heated samples was measured by UK NEQAS using the Trinity Biotech 345 kit (Trinity Biotech) and a Jenway 6715 UV/Vis spectrophotometer with external temperature control to monitor enzyme kinetics and linearity over a set time. Heat-treated material was also assayed for G6PD activity and assessed for its acceptability as EQA survey material by selected UK laboratories. Blood heated at 45 °C for 15 h showed a reduction in G6PD activity of 76.3 ± 4.6% (n = 6) and was considered acceptable as EQA material in terms of appearance and behaviour by the majority of UK sites in the trial. We have developed a simple heat-treatment procedure to produce EQA survey material with low/intermediate G6PD activity, similar to that found in females heterozygous for G6PD deficiency.


Effect of the environment on horizontal gene transfer between bacteria and archaea.
Horizontal gene transfer, the transfer and incorporation of genetic material between different species of organisms, has an important but poorly quantified role in the adaptation of microbes to their environment. Previous work has shown that genome size and the number of horizontally transferred genes are strongly correlated. Here we consider how genome size confuses the quantification of horizontal gene transfer because the number of genes an organism accumulates over time depends on its evolutionary history and ecological context (e.g., the nutrient regime for which it is adapted). We investigated horizontal gene transfer between archaea and bacteria by first counting reciprocal BLAST hits among 448 bacterial and 57 archaeal genomes to find shared genes. Then we used the DarkHorse algorithm, a probability-based, lineage-weighted method (Podell & Gaasterland, 2007), to identify potential horizontally transferred genes among these shared genes. By removing the effect of genome size in the bacteria, we have identified bacteria with unusually large numbers of shared genes with archaea for their genome size. Interestingly, archaea and bacteria that live in anaerobic and/or high temperature conditions are more likely to share unusually large numbers of genes. However, high salt was not found to significantly affect the numbers of shared genes. Numbers of shared (genome size-corrected, reciprocal BLAST hits) and transferred genes (identified by DarkHorse) were strongly correlated. Thus archaea and bacteria that live in anaerobic and/or high temperature conditions are more likely to share horizontally transferred genes. These horizontally transferred genes are over-represented by genes involved in energy conversion as well as the transport and metabolism of inorganic ions and amino acids. Anaerobic and thermophilic bacteria share unusually large numbers of genes with archaea. This is mainly due to horizontal gene transfer of genes from the archaea to the bacteria. In general, these transfers are from archaea that live in similar oxygen and temperature conditions as the bacteria that receive the genes. Potential hotspots of horizontal gene transfer between archaea and bacteria include hot springs, marine sediments, and oil wells. Cold spots for horizontal transfer included dilute, aerobic, mesophilic environments such as marine and freshwater surface waters.


Electrophysiological characterization of retinal Müller glial cells from mouse during postnatal development: comparison with rabbit cells.
The electrophysiology of murine Müller cells, and of their precursors, during postnatal development was investigated by using the whole-cell patch-clamp technique. Membrane potential, membrane capacitance, and expression of voltage-gated Na+ currents increased during the first 3 postnatal weeks. During the same period, the membrane resistance decreased due to an upregulation of both inward and outward K+ currents. Glutamate transporter-mediated currents increased during the first postnatal weeks, as well. In Müller (precursor) cells from rabbits, these transporter currents already achieved adult levels at postnatal day 6, the earliest stage studied. Together with the developmental time course of K+ currents, this indicates a delay in the maturation of murine Müller cells, as compared with cells from rabbit.


The Use of the Overstitch for Bariatric Weight Loss.
Obesity is a public health pandemic and leading contributor to morbidity and mortality. Endoscopic bariatric therapies have emerged as a viable minimally invasive treatment option to fill the therapeutic gap between conservative and surgical approaches. The ability to reliably place full-thickness sutures throughout the gastrointestinal tract has opened the door to novel endoscopic gastric restrictive procedures. A growing body of literature supports endoscopic sleeve gastroplasty as a safe, effective, and reproducible endoscopic treatment of obesity and metabolic syndrome. In addition, endoscopic sutured revision procedures following gastric bypass and sleeve gastrectomy are now first-line with demonstrable safety and long-term efficacy.


The pro-environmental behavioral intention of villagers in rural tourist destinations under China's environmental remediation policy.
This study examined villagers' intention of pro-environmental behavior while supporting the Landcare Policy in China. The research team conducted field surveys of villagers from four famous scenic spots of Cili, which is near the world natural heritage site of the Zhangjiajie natural landscape core area. This area has developed rural tourism, many local villagers rely on tourism to obtain their livelihood income. However, the area is now affected by the environmental remediation policy called Landcare Policy. Cultivated land near the tourist area needs to be repaired, which affects the tourism income of some local villagers. Therefore, local villagers are facing a contradiction between tourism development and environmental protection. The study chose the change in local villagers' pro-environmental intention as the research content. Then we adopted an empirically validated norm activation model (NAM) from Schwartz, and merged the NAM with the expectancy theory of Vroom, based on 511 valid responses from the field questionnaire surveys, we aimed to develop a theoretical framework for researchers to understand the change in villagers' pro-environmental behaviors, concerning the balance between rural tourism livelihood benefits and environmental remediation behavior. Structural equation modeling was conducted for each index of the responses, the findings showed that the merged model had 76.46% better predictive accuracy of villagers' pro-environmental intentions than applying Schwartz's NAM independently. This study found that the motivational force of this new theory significantly influences environmental personal norms due to the joint impact of valence, instrumentality, and expectancy. Villagers with a positive pro-environmental behavior intention expect good tourism benefits and environmental living conditions under the impact of the Landcare policy in rural tourism destinations near the famous natural heritage site.


Graft occlusion following aortofemoral bypass for peripheral ischaemia.
Over a 10-year period, 241 patients with non-aneurysmal aortoiliac disease underwent aortofemoral bypass to 476 limbs. Four patients (1.7 per cent) occluded their grafts within 30 days of surgery, while 25 (10.4 per cent) suffered late graft occlusion. Postoperative occlusions were associated with significant morbidity and only one patient avoided major limb amputation or death. Overall, 46 episodes of graft thrombosis involving 51 graft limbs were encountered, the most common underlying cause being pre-existing or progressive multilevel distal occlusive disease. The overall cumulative graft patency rates were 95 and 87 per cent at 1 and 5 years respectively. Cumulative 5-year patency was significantly higher in patients presenting with claudication (91 per cent) than in patients presenting with rest pain (77 per cent) or ulceration and/or gangrene (71 per cent). Patients with evidence of multilevel distal occlusive disease at the time of aortic surgery had a significantly higher incidence of occlusion compared with those in whom there was no significant distal disease. In 35 episodes of occlusion (76 per cent), surgery was undertaken to restore limb blood flow, being successful in all but one case, with the most commonly performed procedure being graft limb thrombectomy. Seven of 28 patients (25 per cent) ultimately required major limb amputation and three patients died as a direct consequence of graft thrombosis.


Accurate lymph node staging is of greater prognostic importance than subclassification of the T2 category for gastric adenocarcinoma.
Examination of 15 or more lymph nodes is necessary for accurate staging of gastric adenocarcinoma. This study examined whether prognostic discrimination is improved by distinguishing between pT2 tumors limited to the muscularis propria (MP) and those extending to subserosa (SS). A single-institution, prospectively maintained database for 1985-2000 was reviewed for patients who had had R0 resection of pT2 gastric carcinoma. There were 161 patients with MP and 201 patients with SS. The prevalence of nodal metastasis was significantly lower with MP than with SS (47% vs. 66%, respectively; P <.001). As compared with MP, SS was associated with a similar prevalence of pN1 (44% vs. 43%) but a significantly higher prevalence of pN2 or pN3 (3% vs. 23%; P <.001). Five-year survival was significantly greater for patients with MP than with SS (64% vs. 49%; P =.005). On multivariate analyses, however, only the pN category and tumor site were independently significant prognostic factors. With accurate nodal staging, patients with MP or SS had similar survival in association with pN0 (90% vs. 86%) or pN1 (56% vs. 44%). pN2 or pN3 were mainly limited to SS tumors and these patients had poor survival (26% and 0%). For pT2 gastric adenocarcinoma, the depth of mural invasion was not an independently significant prognostic factor.


Evidence for S284L mutation of the CHRNA4 in a white family with autosomal dominant nocturnal frontal lobe epilepsy.
To identify mutations of the neuronal nicotinic acetylcholine receptor alpha4 subunit gene (CHRNA4) responsible for autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) in a group of white patients. A group of 47 patients from 21 unrelated families with ADNFLE were screened for mutations in CHRNA4. Clinical features and EEG findings in the patients were consistent with those reported in the literature for other affected families. The entire gene was amplified from genomic DNA by polymerase chain reaction (PCR) followed by multitemperature single-strand conformation polymorphism analysis (MSSCP) and sequencing. A c.851C>T transition in exon 5 of CHRNA4 was identified in three affected individuals from two generations of the same family, but not in the remaining patients or in 100 healthy volunteers. This mutation caused an S284L substitution in the transmembrane domain M2 segment of the alpha4 subunit of the neuronal nicotinic acetylcholine receptor. The same mutation had previously been detected in a single Japanese family with ADNFLE, and in an Australian woman with a sporadic form of NFLE. This is the first report of an occurrence of c.851C>T transition in a white family with ADNFLE.


Combination extracorporeal shock wave lithotripsy and percutaneous extraction of calculi in a renal allograft.
Renal calculi are a well documented although uncommon complication of kidney transplantation and may be associated with significant morbidity in this immunosuppressed population with a single functioning kidney. We describe a patient who presented with 2 episodes of staphylococcal bacteremia associated with a ureteral structure and struvite calculi involving the calices, renal pelvis and proximal ureter of a cadaveric renal allograft. The patient was treated successfully with a combination of extracorporeal shock wave lithotripsy, percutaneous extraction and balloon dilation of the ureteral stricture. Renal transplant function was not altered postoperatively. In selected cases shock wave lithotripsy can be used as effective adjunctive therapy in a renal allograft harboring stones.


Screening for cervical neoplasia in an unselected rural Guatemalan population using direct visual inspection after acetic acid application: a pilot study.
To assess the acceptability of cervical screening using direct visual inspection after acetic acid application followed by immediate cryotherapy for cervical intraepithelial neoplasia among women in rural Guatemala. An unselected group of 1,052 women voluntarily registered to undergo cervical screening using direct visual inspection of the cervix after acetic acid application. Women with acetowhite changes consistent with cervical intraepithelial neoplasia were offered immediate cryotherapy. Cervical screening was deferred in 80 (7.6%) registrants, and 18 (1.7%) refused to undergo an examination. Among the 954 registrants screened, 125 (13%) had findings consistent with cervical intraepithelial neoplasia. Cryotherapy was deferred in three patients. A total of 121 (99%) women agreed to immediate cryotherapy. Direct cervical visualization after acetic acid application followed by immediate cryotherapy for acetowhite changes consistent with cervical intraepithelial neoplasia would be a well-accepted method of cervical screening in rural Guatemala.


Irreducible trans-scaphoid perilunate dislocation.
Two cases of irreducible trans-scaphoid perilunate dislocation are described. In each case there was soft tissue interposition between the proximal and distal elements of the fracture-dislocation and this prevented capitolunate and scaphoid reduction. The dorsal capsule was impaled on the distal scaphoid fragment in one case, and a flap of dorsal capsule was entrapped between the capitate and the lunate in the other. In light of these findings, we recommend a dorsal approach for open reduction of trans-scaphoid perilunate dislocation when the capitolunate joint cannot be reduced by closed manipulation.


Cell differentiation in the longitudinal veins and formation of commissural veins in rice (Oryza sativa) and maize (Zea mays).
Vascular development is a central theme in plant science. However, little is known about the mechanism of vascular development in monocotyledons (compared with dicotyledons). Therefore, we investigated sequential processes of differentiation into various different vascular cells by carrying out detailed observations using serial sections of the bases of developing leaves of rice and maize. The developmental process of the longitudinal vascular bundles was divided into six stages in rice and five stages in maize. The initiation of differentiation into procambial progenitor cells forming the commissural vein arose in a circular layer cell that was adjacent to both a metaxylem vessel and one or a few phloem cells in stage V longitudinal vascular bundles. In most cases the differentiation of ground meristem cells into procambial progenitor cells extended in one direction, toward the next longitudinal vascular bundle, and subsequent periclinal divisions and further differentiation produced a vessel element, two companion cells and a sieve element to form a commissural vein. These results suggest the presence of an intercellular signal(s) that induces differentiation of the circular layer cell and the ground meristem cells into procambial progenitor cells, forming a commissural vein sequentially.


Adaptive natural killer cell response to cytomegalovirus and disability progression in multiple sclerosis.
Human cytomegalovirus (HCMV) causes a highly prevalent infection which may have a multifaceted impact on chronic inflammatory disorders. However, its potential influence in multiple sclerosis (MS) remains controversial. The HCMV-host interaction may induce an adaptive reconfiguration of the natural killer (NK) cell compartment, whose hallmark is a persistent expansion of peripheral NKG2C+ NK-cells. The purpose of this study was to evaluate whether the HCMV-driven NKG2C+ NK-cell expansion is related to the MS clinical course. Multicentre analysis of NKG2C expression and genotype according to HCMV serostatus and time of assignment of irreversible disability scores in 246 MS patients prospectively followed up in our institutions. NKG2C expression was unrelated to disease-modifying drugs, remained stable under steady-state conditions, and was higher in HCMV(+) NKG2C(+/+) homozygous individuals. NKG2C+ NK-cell expansion in HCMV(+) patients, as compared to HCMV(+) or HCMV(-) patients with lower NKG2C+ NK-cells proportions, conferred a lower risk of progression in Cox regression analysis (Expanded Disability Status Scale (EDSS)>3.0, hazard ratio (HR)=0.33, 95% confidence interval (CI) 0.15-0.71, p=0.005; EDSS>5.5, HR=0.23, 95% CI 0.07-0.74, p=0.014). Neither HCMV serostatus nor NKG2C genotype appeared to be related to disability progression. HCMV may exert a beneficial influence on MS, decreasing the risk of disability progression in those patients displaying a virus-driven NKG2C+ NK-cell expansion.


Synthesis and characterization of novel natural product-Gd(III) MRI contrast agent conjugates.
Several novel gadolinium chelates conjugated with paclitaxel, colchicine and thyroxine have been prepared as MRI contrast agents targeted to tubulin and thyroxine-binding globulin, respectively.


Molecular investigation of tick-borne pathogens in ticks from grazing cattle in Korea.
This study was carried out to identify the tick species that infest grazing cattle and to determine the presence of tick-borne pathogens transmitted by these ticks in Korea. A total of 903 ticks (categorized into 566 tick pools) were collected from five provinces during 2010-2011. The most prevalent tick species was Haemaphysalis longicornis, followed by three Ixodes spp. ticks. The collected ticks were infected with both rickettsial and protozoan pathogens. In all, 469 (82.9%) tick pools tested positive for the Anaplasma/Ehrlichia 16S rRNA gene, whereas 67 (11.8%) were positive for the Babesia/Theileria 18S rRNA gene. Among the rickettsial pathogens, E. canis was detected with the highest rate (22.3%), followed by A. platys (20%), E. chaffeensis (19.4%), E. ewingii (19.3%), Rickettsia sp. (12.4%), A. phagocytophilum (5.5%) and E. muris (0.5%). Among the protozoan pathogens, T. equi was detected with the highest rate (7.2%), followed by T. sergenti/T. buffeli (3.7%) and B. caballi (0.35%). Simultaneous infections with up to seven pathogens were also identified. In particular, ticks infected with rickettsial pathogens were also infected with protozoan pathogens (22 samples). All five provinces investigated infected with tick-borne pathogens.


Micro-structural, linear and nonlinear optical properties of titania nanoparticles.
TiO(2) (titania) nanoparticles were prepared by sol-gel method using titanium tetrachloride as precursor and calcined at 500°C (SA1) and 800°C (SA2). X-ray diffraction pattern of SA1 has mixed phases of anatase (17%) and rutile (83%) whereas SA2 has pure rutile (100%) TiO(2). From the X-ray diffraction peak profile analysis, the crystallite size, lattice strain, deformation stress and energy density value of TiO(2)nanoparticles were determined more precisely by Williamson-Hall method, uniform deformation model, uniform stress deformation model, uniform deformation energy density model and size-strain plot method. The calculated average crystallite size from uniform stress deformation model is 21 and 53nm for SA1 and SA2 respectively. From the FTIR spectra Ti-O vibrational frequency is observed at ∼530cm(-1). Using Tauc's relation the direct bandgap of SA1 and SA2 is calculated from UV-vis spectra as 2.75 and 2.95eV respectively. The excitation wavelength dependent photoluminescence has been investigated for 290 and 330nm as excitation wavelengths. It was observed that the band edge emission is decreased with respect to excitation wavelengths. The second harmonic generation efficiency (SHG) observed for centrosymmetric rutile TiO(2) is more comparable with KDP crystal.


Magnetic resonance microscopy atlas of equine embryonic development.
Equine embryogenesis post implantation is not well studied, and only two-dimensional illustrations are available. A thorough appreciation of the complex three-dimensional relationship between tissues and organs and their development is, however, crucial for understanding physiological and pathological mechanisms. The goals were 2-fold: 1) to establish a freely accessible online atlas as a reference tool for the scientific and pedagogic communities; and 2) to create a framework for integration of data with known spatiotemporal distribution, such as gene expression or cell lineage. Descriptive anatomical study. Magnetic resonance microscopy was performed on embryos of 28, 32, 35, 37, 39, 40, 42, 45, 50 and 65 days gestation using a 9.4 T magnet. Equine embryos were staged according to the Carnegie system. Acquired images were optimised using histogram optimisation and processed for easy online access. Magnetic resonance microscopy protocols for imaging of equine embryos and fetuses were developed. The wider spread of signal intensity values achieved by histogram equalisation increased visual contrast considerably. Despite their longer gestation, equine conceptuses appeared to reach the various Carnegie staging benchmarks earlier than human embryos. The equine atlas is designed to serve as an online reference tool for research and teaching. The equine atlas may serve as a foundation and scaffold for improved anatomical labelling, spatial and temporal data integration and further understanding of physiological and pathophysiological processes involved in development and disease.


Predicting recurrence after radiotherapy in head and neck cancer.
Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide. Radiotherapy is a mainstay of treatment, either alone for early stage tumors or combined with chemotherapy for late stage tumors. An overall 5-year survival rate of around 50% for HNSCC demonstrates that treatment is often unsuccessful. Prediction of outcome is, therefore, aimed at sparing patients from ineffective and toxic treatments on the one hand, and indicating more successful treatment modalities on the other. Both functional and genetic assays have been developed to predict intrinsic radiosensitivity, hypoxia, and repopulation rate. Few, however, have shown consistent correlations with outcome across multiple studies. Messenger RNA and microRNA profiling show promise for predicting hypoxia, whereas epidermal growth factor receptor expression combined with other measures of tumor differentiation grade shows promise for predicting repopulation rate. Intrinsic radiosensitivity assays have not proven useful to date, although development of repair protein foci assays indicates promise from preclinical studies. Assays for cancer stem cell content have shown promise in several clinical studies. In addition, 2 assays showing robustness as predictors for outcome in HNSCC are human papilloma virus status and epidermal growth factor receptor expression. Neither these nor stem cell assays, however, can as yet reliably indicate alternative and better treatments for poor prognosis patients. It would be of great value to have assays that predict the benefit for an individual from combining new molecularly targeted agents with radiotherapy to increase response, in particular those that exploit tumor mutations to provide tumor specificity. Predictive assays are being developed for detecting defects in repair pathways for single- and double-strand DNA breaks, which should allow selection of drugs targeting the appropriate backup pathway, thus exploiting the concept of synthetic lethality. This is one of the most promising areas for prediction, both currently and in the future.


The assembly of regularly spaced nucleosomes in the Xenopus oocyte S-150 extract is accompanied by deacetylation of histone H4.
Histone proteins, which were assembled into chromatin using the Xenopus oocyte S-150 extract, were analyzed on acid-urea gels and Triton-acid-urea gels to determine their state of modification. We find that histone H4, which is present in a diacetylated form in the oocyte S-150, gradually loses its acetate groups as the DNA is packaged into chromatin. Thus, this process parallels the one observed in vivo during chromatin formation in growing eucaryotic cells. Histone H4 deacetylation in the oocyte S-150 is a DNA-dependent reaction. This reaction is blocked when butyrate (an inhibitor of histone deacetylase) is added at the onset of the chromatin assembly process. When butyrate is added at the end of the assembly process, no de novo acetylation of the nucleosomal histone H4 is observed. Chromatin with regularly spaced nucleosomes, displaying periodicities ranging from 160 to 220 base pairs, can be assembled in vitro with the oocyte S-150 (Rodríguez-Campos, A., Shimamura, A., and Worcel, A. (1989) J. Mol. Biol., in press). This chromatin may contain either deacetylated histone H4 when assembled under standard conditions or diacetylated H4 when assembled in the presence of butyrate. Both types of chromatin display identical structures upon digestion with nucleases. The potential applications of this system toward the study of the naturally occurring diacetylated histone H4 are discussed.


Laser fluorimetry: subpicogram detection of aflatoxins using high-pressure liquid chromatography.
The use of high-pressure liquid chromatographic separation in conjunction with laser-induced fluorescence detection permits the analysis of trace fluorescent species at new limits of sensitivity. This technique was applied to the carcinogens aflatoxins B1, B2, G1, and G2, which were linearly quantitated to 7.5 x 10(-13) gram. The procedure consists of forming more fluorescent aflatoxin derivatives, eluting the aflatoxins from a reverse-phase column, focusing the 325-nanometer output of a helium-cadmium ion laser into a suspended droplet of the eluent, and measuring the resulting fluorescence using phase-sensitive detection.


Experimental and Quantum Chemical Studies of Nicotinamide-Oxalic Acid Salt: Hydrogen Bonding, AIM and NBO Analysis.
The computational modeling supported with experimental results can explain the overall structural packing by predicting the hydrogen bond interactions present in any cocrystals (active pharmaceutical ingredients + coformer) as well as salts. In this context, the hydrogen bonding synthons, physiochemical properties (chemical reactivity and stability), and drug-likeliness behavior of proposed nicotinamide-oxalic acid (NIC-OXA) salt have been reported by using vibrational spectroscopic signatures (IR and Raman spectra) and quantum chemical calculations. The NIC-OXA salt was prepared by reactive crystallization method. X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC) techniques were used for the characterization and validation of NIC-OXA salt. The spectroscopic signatures revealed that (N7-H8)/(N23-H24) of the pyridine ring of NIC, (C═O), and (C-O) groups of OXA were forming the intermolecular hydrogen bonding (N-H⋯O-C), (C-H⋯O═C), and (N-H⋯O═C), respectively, in NIC-OXA salt. Additionally, the quantum theory of atoms in molecules (QTAIM) showed that (C10-H22⋯O1) and (C26-H38⋯O4) are two unconventional hydrogen bonds present in NIC-OXA salt. Also, the natural bond orbital analysis was performed to find the charge transfer interactions and revealed the strongest hydrogen bonds (N7-H8⋯O5)/(N23-H24⋯O2) in NIC-OXA salt. The frontier molecular orbital (FMO) analysis suggested more reactivity and less stability of NIC-OXA salt in comparison to NIC-CA cocrystal and NIC. The global and local reactivity descriptors calculated and predicted that NIC-OXA salt is softer than NIC-CA cocrystal and NIC. From MESP of NIC-OXA salt, it is clear that electrophilic (N7-H8)/(N23-H24), (C6═O4)/(C3═O1) and nucleophilic (C10-H22)/(C26-H38), (C6-O5)/(C3-O2) reactive groups in NIC and OXA, respectively, neutralize after the formation of NIC-OXA salt, confirming the presence of hydrogen bonding interactions (N7-H8⋯O5-C6) and (N23-H24⋯O2-C3). Lipinski's rule was applied to check the activeness of salt as an orally active form. The results shed light on several features of NIC-OXA salt that can further lead to the improvement in the physicochemical properties of NIC.


The complete nucleotide sequences of the Escherichia coli LIV-BP and LS-BP genes. Implications for the mechanism of high-affinity branched-chain amino acid transport.
The Escherichia coli LIV-I and LS amino acid transport systems are high-affinity, periplasmic, binding protein-dependent systems that utilize the leucine-, isoleucine-, valine-binding protein (LIV-BP) and leucine-specific binding protein (LS-BP), respectively. These two binding proteins (BPs) interact with a common set of membrane proteins to transport branched-chain amino acids into the cytoplasm. The two BP genes are encoded in a regulon at minute 76 of the E. coli chromosome that also contains the genes for the common membrane protein components. We report here the nucleotide and deduced amino acid sequences for the LIV-BP and LS-BP genes and their protein products. Both BPs are encoded with a 23-amino acid signal sequence that is removed when the BPs are secreted into the periplasm. We have examined the pattern of amino acid sequence conservation between the two BP molecules and, by comparison of the predicted secondary structures to the 2.0-A crystal structure of the LIV-BP, have found regions of the BPs that may be involved in membrane protein interaction. We also analyzed the translational efficiency of the two BP mRNAs as determined by their nucleotide sequences.


Catabolism of myo-inositol to precursors utilized for de novo glycerolipid biosynthesis.
Systemic injection of [2-3H]myo-inositol into frogs resulted in the incorporation of more than half of the label into glycerolipid classes other than phosphoinositides in retinal rod outer segment membranes. Following methanolysis and differential extraction of isolated lipid classes, radioactivity was recovered primarily in the aqueous phase. After phospholipase C hydrolysis of the total membrane lipids, 97% of the radioactivity was extractable with organic solvents, and 70% of the label in lipids was in 1,2-diglycerides. These results indicate that the label was incorporated primarily into the glyceryl moiety of the membrane glycerolipids. Intraocular injection of frog eyes or in vitro incubation of frog retinas with [2-3H]myo-inositol resulted in the incorporation of radioactivity almost exclusively into phosphoinositides in rod outer segment membranes. Incubation of retinas with [U-14C]glucuronic acid did not result in the formation of labeled retinal lipids. These results suggest that myo-inositol can be catabolized systemically to precursors utilized for glycerolipid biosynthesis in the retina.


Is Age ≥ 55 years an optimal cutoff point among patients with differentiated thyroid Microcarcinoma? a preliminary study.
Age ≥ 55 years is regarded as a pivotal component of TNM stage classification in differentiated thyroid carcinoma (DTC). However, whether this cutoff point is still adaptable for differentiated thyroid microcarcinoma (DTMC) is rarely investigated. We reviewed and analyzed the data of DTC patients aged ≥ 55 years from the Surveillance, Epidemiology, and End Results (SEER) database. Univariate and multivariate Cox regression analyses were used to determine the potential risk factors of cancer-specific survival (CSS) in DTMC patients aged ≥ 55 years. The Kaplan-Meier survival curves were used to estimate CSS probability. Receiver operating characteristic (ROC) curves were used to analyze the best age cutoff point for DTMC. Among the DTMC patients, there was no significant difference in the 1-, 3-, 5-, and 7-year CSS probability between the 55-59 years and 60-64-years subgroup (p = 0.72). The ROC curves indicated that 65 years, 65 years, and 64 years were the cutoff age point of 3-, 5-, and 7-year CSS probability in DTMC patients, respectively. Besides, N1b (Hazard ratio (HR) = 3.90, 95% Confidence interval (CI): 2.01-7.57; p < 0.001), extrathyroidal extension (HR = 2.53, 95 %CI: 1.39-4.62; p = 0.002), and M1 (HR = 11.42, 95 %CI: 5.04-25.90; p < 0.001) were the independent risk factors in CSS of DTMC patients. Our results suggested age at diagnosis ≥ 55 years is not the best cutoff point in stratifying the stage of the DTMC patients. On the contrary, those patients aged above 65 years have a significantly lower probability of CSS, which perhaps should be taken into consideration for treatment decision-making.


Development and evaluation of an inhibitor-releasing matrix for intrauterine devices.
Antifibrinolytic agents when released into the uterine cavity decrease menorrhagia associated with IUD use. Our objective was to develop a matrix that could be incorporated onto an IUD and release anti-fibrinolytic agents. The copolymer ethylene-vinyl acetate (EVA) was selected for detailed study because it has the advantage over other materials in that it can release large molecular weight substances for more than 100 days, and allows incorporation of large amounts of anti-fibrinolytic agents with different molecular weights. Two compounds, tranexamic acid (AMCA, MW=157) and Trasylol (Kunitz pancreatic trypsin inhibitor, (MW=6,500) were incorporated into the EVA matrix and their release rates measured. In vitro studies with AMCA showed that after the initial burst, a constant high release rate was obtained over a prolonged period of time. The in utero release rate of AMCA from the EVA matrix in rabbits was similar to that obtained in vitro. By contrast, the release rate of Trasylol decreased to low levels during incubation in vitro. The release rate of Trasylol in utero however, appeared to be higher than that in vitro.


Balloon catheter vs oxytocin alone for induction of labor in women with one previous cesarean section and an unfavorable cervix: a multicenter, retrospective study.
To compare the rate of vaginal birth between double-balloon catheter and oxytocin alone for induction of labor in women with one previous cesarean section and an unfavorable cervix. A retrospective and observational study was conducted from 2013 to 2017, at the Saint-Etienne University Hospital where women received induction with a double-balloon catheter for 12 h and at the Grenoble Alpes University Hospital where women received induction with a low-dose oxytocin infusion. Primary outcome was the rate of vaginal birth. Out of 1920 women eligible for attempting a vaginal birth after one previous cesarean section, 501 had a labor induction. Among women with an unfavorable cervix, 160 received a double-balloon catheter in Saint Etienne and 152 received oxytocin alone in Grenoble. The vaginal birth rate was higher in the double-balloon catheter group (61% versus 47% in the oxytocin group). An induction of labor with oxytocin alone reduced chances of vaginal birth (aOR 0.38 CI-95% [0.22-0.66]) compared to cervical ripening with double-balloon catheter. The perinatal morbidity was similar in the two groups. There was, however, 3.9% uterine rupture in the oxytocin group versus 0.6% in the double-balloon group (p = 0.11). For induction of labor in women with one previous cesarean section and with unfavorable cervix, cervical ripening with a double-balloon catheter increases the rate of vaginal birth without increased risk of uterine rupture.


Patient knowledge and preferences for peripheral artery disease treatment.
Shared medical decision making requires patients' understanding of their disease and its treatment options. Peripheral artery disease (PAD) is a condition for which preference-sensitive treatments are available, but for which little is known about patients' knowledge and treatment preferences as it relates to specific treatment goals. In a prospective, multicenter registry that involved patients with PAD experiencing claudication, the PORTRAIT Knowledge and Preferences Survey was administered at 1 year. It asks questions about PAD treatment choices, symptom relief options, disease management, and secondary prevention. PAD treatment preferences were also queried, and patients ranked 10 PAD treatment goals (1-10 Likert scale; 10 being most important). Among 281 participants completing the survey (44.8% women, mean age 69.6 ± 9.0 years), 54.1% knew that there was more than one way to treat PAD symptoms and 47.1% were offered more than one treatment option. Most (82.4%) acknowledged that they had to manage their PAD for the rest of their life. 'Avoid loss of toes or legs,' 'decreased risk of heart attack/stroke,' 'long-lasting treatment benefit,' 'living longer,' 'improved quality of life,' and 'doing what the doctor thinks I should do' had mean ratings > 9.0 (SD ranging between 1.21 and 2.00). More variability occurred for 'avoiding surgery.' 'cost of treatment,' 'timeline of pain relief,' and 'return to work' (SD ranging between 2.76 and 3.58). The single most important treatment goal was 'improving quality of life' (31.3%). Gaps exist in knowledge for patients with PAD who experience claudication, and there is a need for increased efforts to improve support for shared decision-making frameworks for symptomatic PAD.(ClinicalTrials.gov Identifier: NCT01419080).


Juxtaposition of the atrial appendages.
Juxtaposition of the atrial appendages is a rare congenital cardiac anomaly almost universally associated with severe conotruncal abnormalities, especially transposition of the aorta. This presentation describes a case of right sided juxtaposition of the atrial appendages without concomitant conotruncal malformations. This finding contradicts previous hypotheses concerning the genesis of juxtaposition.


Extraction, partial purification and characterization of proteases from the red seaweed Gracilaria edulis with similar cleavage sites on κ-casein as calf rennet.
Enzymes currently used in cheesemaking have various drawbacks, and there is a continual need to find new coagulants. This study describes the extraction and biochemical characterization of two proteases from the red alga Gracilaria edulis. The proteases were extracted with phosphate buffer and partially purified by ammonium sulphate precipitation and dialysis. The enzymes exhibited optimum caseinolytic activity at 60 °C and a pH range of 6-8. They showed a high ratio of milk-clotting over caseinolytic activity, indicating they had an excellent milk-clotting ability. The proteases were confirmed to be serine protease and metalloprotease with molecular weight (MW) of 44 and 108 kDa. They exhibited high hydrolytic activity on κ-caseins, cleaving κ-casein at four main sites, one of which being the same as that of calf rennet, which is the first reported for an algal protease. The findings demonstrated that the proteases could potentially be used as a milk coagulant in cheesemaking.


Identification and characterization of long non-coding RNAs in intestinal immune regulation of largemouth bass, Micropterus salmoides, under acute heat stress.
Long noncoding RNAs (lncRNAs), which are RNA molecules that do not code for proteins and have a length exceeding 200 base pairs, have been found to play a crucial role in regulating intestinal immunity. The high mortality of various fish species induced by high temperatures is known to be associated with enteritis. Our investigation demonstrated that acute heat stress was responsible for inducing fish enteritis. However, the specific lncRNAs involved this process remains unknown. In this current study, we utilized intestinal sequencing data from the largemouth bass species Micropterus salmoides under acute heat stress, resulting in a total of 347,351,492 clean reads obtained from six cDNA libraries. A total of 3399 novel lncRNA transcripts originating from 2488 distinct lncRNA genes were successfully identified. Consistent with previous findings in other fish species, these lncRNAs demonstrated comparatively shorter transcript lengths when compared to protein-coding genes. Furthermore, a total of 216 novel lncRNA exhibited differential expression (DE) in the intestine of largemouth bass, meeting the criteria of absolute log2 fold change exceeding 2 and a p-value below 0.05. Additionally, these DE-lncRNAs were found to regulate 210 neighboring genes in a cis-regulatory manner. An examination of GO/KEGG enrichment revealed a notable enrichment of immune regulation (p < 0.05) among these cis-genes, with lncRNA MSTRG.8573.1 playing a significant role in regulating the jak-stat signaling pathway during this process. This study presents a comprehensive inventory of novel DE-lncRNA implicated in the development of enteritis in largemouth bass under acute heat stress. These findings offer valuable insights for future investigations on the regulation of lncRNAs to mitigate heat stress-induced fish enteritis.


Clinical significance of Her-2/neu overexpression in uterine serous carcinoma.
To evaluate the clinico-pathologic characteristics and survival outcome associated with overexpression of Her-2/neu in patients with uterine serous carcinoma (USC). Twenty-five patients with a confirmed pathologic diagnosis of USC and available paraffin embedded tissue samples treated at the Johns Hopkins Medical Institutions from 1/1/1992 through 12/31/2000 were identified retrospectively. Her-2/neu expression was evaluated by immunohistochemistry using HercepTest (DAKO). Clinical data were abstracted from medical records. Clinico-pathologic characteristics associated with Her-2/neu overexpression like staging, histology, lymph-vascular space involvement, and myometrial invasion were evaluated using logistic regression analysis and Fisher's exact test. Analyses of overall survival time were performed using the Kaplan-Meier method and Cox proportional hazards regression models. Twelve (48%) of the 25 USC cases demonstrated Her-2/neu overexpression. There was a significant difference in Her-2/neu overexpression and surgical staging (81.8% vs. 28.6%, P = 0.01). Survival analysis according to primary tumor characteristics revealed that overexpression of Her-2/neu was significantly associated with a worse survival outcome (HR = 6.58, 95%CI: 1.36-31.89, P = 0.02). Her-2/neu overexpression is associated with advanced surgical stage USC and poor survival outcome. These data may be useful in guiding the clinical management of patients with USC and have potential implications for the development of novel treatment strategies.


Laser-Induced Ocular Hypertension in a Mouse Model of Glaucoma.
Glaucoma is a neurodegenerative disease that leads to the loss of retinal ganglion cells (RGC) and thus to blindness. There are numerous experimental models used for the study of this pathology. Among the different models, episcleral vein photocoagulation is one of the most widely used. In this model there is a transient increase in intraocular pressure that returns to normal values about 7 days after induction of ocular hypertension (OHT). In addition, typical glaucoma changes, such as loss of RGC, thinning of the optic nerve fiber layer, and glial activation, occur in this model. All these changes have been described in detail over time after OHT induction. In this chapter, we describe the detailed method of OHT induction in Swiss albino mice by diode laser photocoagulation of limbal and episcleral veins.


Primary intracranial tumours in Black and Indian children, 1960-1975.
This report of cerebral tumours in 60 children admitted to the medical wards of King Edward VIII Hospital, Durban, shows that cerebral tumour is the commonest solid neoplasm in both Black and Indian children. There is a significantly lower incidence of cerebral tumour in Black children. No difference was apparent in age, sex ratio, site or histological types in our racial groups compared with studies in White children from other parts of the world.


Habitual coffee consumption and blood pressure: A study of self-defense officials in Japan.
The study aims to examine the relationship between habitual coffee consumption and blood pressure. The subjects were 3336 male self-defense officials aged 48-56 years, who received a preretirement health examination at the Self-Defense Forces Fukuoka Hospital between October 1986 and December 1992. Average coffee intake in the past year was ascertained by a self-administered questionnaire. A significant inverse relation between habitual coffee consumption and blood pressure was found with and without adjustment for alcohol use, cigarette smoking, body mass index, glucose tolerance, and green tea intake. Green tea, another major source of caffeine intake in Japanese, was unrelated to blood pressure. The adjusted mean differences per cup of coffee consumed per day were -0.6 mmHg (95% confident interval [CI]: -0.9 to -0.3, p = 0.0001) in systolic blood pressure and -0.4 mmHg (95% CI: -0.5 to -0.2, p = 0.0002) in diastolic blood pressure. Habitual coffee drinkers had lower blood pressure than non-drinkers at any levels of alcohol use, cigarette smoking, obesity, and glucose intolerance. Our findings consolidate the previous observation that habitual coffee consumption was associated with lower blood pressure.


Microangiographic imaging of the prostate.
To present the results of microangiography in visualizing prostatic microvascularization. A contrast medium was injected into the vessels of prostate specimens obtained during surgery, to fill small vessels; nine specimens were obtained after cystoprostatectomy and two after radical prostatectomy. Branches of prostatic or seminal vesicle arteries were used for the injection, the specimens then fixed in formalin, sectioned and X-rayed. In five samples the quality of the images was sufficient to visualize small arteries; the remaining vessels were partially filled. The vascular map was compared with previous results from historadiographic studies. Microangiographic imaging of prostate specimens provides a unique view of the angioarchitecture of the prostate and correlates closely with historadiographic studies. The correlation with other imaging methods, e.g. Doppler ultrasonography, needs further evaluation.


Phase II study on cisplatin and ifosfamide in recurrent high grade gliomas.
27 patients with recurrent high grade glioma following surgery and radiation therapy were treated with 100 mg/m2 cisplatin and 6 g/m2 ifosfamide per cycle, administered on days 1-3 in 4 week cycles, for a maximum of six cycles. Toxicity was assessed after every cycle. Response was assessed following every second cycle, and a 50% decrease of the largest cross-sectional tumour area on contrast enhanced magnetic resonance imaging or computed tomography scan was considered a partial response (PR). A total of 95 cycles was administered; 26 patients were evaluable for response. In 5 patients (19%), a PR was obtained (median time to progression (TTP): 34 weeks). Stable disease was observed in 6 patients (23%, median TTP: 22 weeks). The most frequent toxicity was haematological: 37% of cycles were complicated by a grade 3 or 4 leucopenia. 1 patients died, probably as a consequence of increased cerebral oedema induced by the cisplatin hydration schedule. Determination of the cisplatin concentration in this patient showed a 10-fold increase in the tumour concentration as compared with that in normal brain tissue, demonstrating the absence of a blood-brain barrier in the tumour. In conclusion, generally this schedule was well tolerated, but it is of moderate activity for recurrent glioma.


Rational strain improvement for surfactin production: enhancing the yield and generating novel structures.
Surfactin, one of the most powerful microbial surfactants, is a lipopeptide-type biosurfactant which combines interesting physicochemical properties and biological activities. However, the high cost caused by its low productivity largely limits the commercial application of surfactin. Hence, many engineered bacterium have also been used to enhance surfactin biosynthesis. This review briefly summarizes the mechanism of surfactin biosynthesis, highlighting the synthesis pathway of N-terminally attached fatty acids, and outlines the main genetic engineering strategies for improving the yield and generating novel structures of surfactin, including promoter engineering, enhancing efflux systems, modifying the transcriptional regulatory genes of surfactin synthase (srfA), genomics and transcriptomics analysis, non ribosomal peptide synthetase (NRPS) domain and combinatorial biosynthesis. Finally, we discuss the future prospects of the research on surfactin.


NIR spectroscopic determination of urine components in spot urine: preliminary investigation towards optical point-of-care test.
Presently, there is no convenient method to measure 24-h urinary Na excretion, which is an important index of daily Na intake, and 24-h urine collection involves a complex process. However, the Na-to-creatinine ratio (NCR) in spot urine has the potential to evaluate 24-h Na excretion and is useful for point-of-care testing. Thus, this study aimed to realize a near-infrared spectroscopic system to assess NCR in spot urine: (1) We attempted to estimate Na concentration using fewer than 10 wavelengths; (2) we calculated NCR using creatinine concentrations from our previous report and verified the NCR predictability. A calibration model was created using multi-linear regression analysis using 10 selected wavelengths in the range of Fourier-transform infrared spectrometer. Spot urine samples were obtained from healthy adults, and glucose powder was added to them to simulate diabetic samples. NCR was calculated using only six wavelengths, and the results confirmed the high accuracy of the estimated Na concentration even though inorganic components do not absorb near-infrared light. Our method enables to optically estimate NCR in spot urine, and it will be useful for point-of-care testing. Graphical abstract.


MAPK11 (p38β) is a major determinant of cellular radiosensitivity by controlling ionizing radiation-associated senescence: An in vitro study.
MAPKs are among the most relevant signalling pathways involved in coordinating cell responses to different stimuli. This group includes p38MAPKs, constituted by 4 different proteins with a high sequence homology: MAPK14 (p38α), MAPK11 (p38β), MAPK12 (p38γ) and MAPK13 (p38δ). Despite their high similarity, each member shows unique expression patterns and even exclusive functions. Thus, analysing protein-specific functions of MAPK members is necessary to unequivocally uncover the roles of this signalling pathway. Here, we investigate the possible role of MAPK11 in the cell response to ionizing radiation (IR). We developed MAPK11/14 knockdown through shRNA and CRISPR interference gene perturbation approaches and analysed the downstream effects on cell responses to ionizing radiation in A549, HCT-116 and MCF-7 cancer cell lines. Specifically, we assessed IR toxicity by clonogenic assays; DNA damage response activity by immunocytochemistry; apoptosis and cell cycle by flow cytometry (Annexin V and propidium iodide, respectively); DNA repair by comet assay; and senescence induction by both X-Gal staining and gene expression of senescence-associated genes by RT-qPCR. Our findings demonstrate a critical role of MAPK11 in the cellular response to IR by controlling the associated senescent phenotype, and without observable effects on DNA damage response, apoptosis, cell cycle or DNA damage repair. Our results highlight MAPK11 as a novel mediator of the cellular response to ionizing radiation through the control exerted onto IR-associated senescence.


Bilateral atypical femoral fracture and end-stage arthritis of the hip, treated with total hip arthroplasty.
The prolonged use of bisphosphonates has been associated with an increased rate of atypical femoral fracture. A 77-year-old woman with prolonged bisphosphonate use presented to our office with groin pain and end-stage arthritis, She was scheduled for a total hip replacement. Before the surgery and with minimal trauma, the patient then suffered a displaced atypical femoral fracture. She underwent a total hip replacement as a treatment for her fracture and her arthritis. Subsequently, the patient presented with pain in the contralateral thigh with an incomplete atypical femoral fracture. That side was also treated with a total hip arthroplasty. An uncemented stem with open reduction internal fixation and a long cemented stem were used on the complete fracture and incomplete fracture sides, respectively. At a follow-up of 2 years, the patient had no pain and had excellent function demonstrating the short-term success of both cemented and uncemented stems in total hip arthroplasty after atypical femoral fractures.


Malaria parasite prevalence and Haematological parameters in HIV seropositive patients attending the regional hospital Limbe, Cameroon: a hospital-based cross-sectional study.
Malaria and the human immunodeficiency virus (HIV) infection constitute public health problems in Cameroon including the South West Region (SWR). This study determined the prevalence of malaria parasites and haematological abnormalities in HIV positive patients in Limbe, Cameroon from April-July 2014. The study was cross-sectional and involved 411 participants who were administered structured questionnaires to record socio-demographic and clinical data. Three hundred and nine (309) HIV positive patients and one hundred and two (102) HIV negative individuals were examined clinically and venous blood collected for malaria parasite detection, HIV infection diagnosis and full blood count analysis. Overall malaria parasite prevalence was 14.1% (58/411). This prevalence was significantly higher (P < 0.001) in the HIV negative participants (33.3%, 34/102) compared to the HIV positive patients (7.8%, 24/309). Amongst HIV positive participants, malaria parasite prevalence was significantly higher in female patients (P = 0.003), febrile patients (P < 0.001), anaemic patients (P = 0.015) and in patients who were not on antiretroviral treatment (ART) (P = 0.03) when compared with their respective counterparts. Among the HIV negative group, though not significant, malaria parasite prevalence was higher in females, febrile and anaemic patients when compared with their respective counterparts. Overall anaemia prevalence was 52.1% (214/309) and was significantly higher (P = 0.004) in HIV positive patients (56%, 173) than in HIV negative participants (40.2%, 41). Malaria/HIV co-infected patients had a significantly lower mean value of Hb (P = 0.002), RBC (P = 0.002) and Hct (P = 0.001) when compared with HIV-infected patients. HIV negative participants had a higher prevalence of malaria parasites than their HIV positive counterparts. Anaemia prevalence was higher in HIV positive patients than in HIV negative participants. Malaria/HIV co-infected patients presented with more red blood cell abnormalities than HIV-infected patients.


Insomnia Symptom Trajectory of Spouse Caregivers of Older Adults with Functional Limitations.
This study examined the long-term impact of spouse caregiving on insomnia symptoms, compared to propensity-score matched non-caregivers. Health and Retirement Study data between 2006 and 2018 were used. Caregivers (n = 403) were respondents (aged 50+) who assisted their heterosexual spouses in performing (instrumental) activities of daily living at baseline. Non-caregivers were matched using a propensity score matching procedure based on baseline characteristics. Insomnia symptoms were measured every 4 years for both groups. Poisson mixed-effect models estimated the association between caregiver status and insomnia symptoms. Compared to matched non-caregivers, caregivers had similar severity of insomnia symptoms at baseline (βcaregiver = 0.018, 95% CI = -0.089, 0.124) and reported a similar yearly change rate (βcaregiver×time = -0.008, 95% CI = -0.017, 0.001). No moderation effects of care-recipients' dementia status and social support were significant. In this study sample, there is no evidence that spouse caregivers, specifically those who performed light duties, experience more severe insomnia symptoms than non-caregivers. Spouse caregiving, especially in a light-duty capacity, may not be detrimental to the caregivers' sleep health. More data are needed regarding insomnia in spouse caregivers with heavy duties of care to fully assess the health impact of the caregiving experience.


Ganciclovir is effective for prophylaxis and treatment of human herpesvirus-6 in allogeneic stem cell transplantation.
Human herpesvirus 6 (HHV-6) infection and disease are serious complications of allogeneic hematopoietic stem cell transplantation (allo-SCT). Ganciclovir (GCV) is effective against HHV-6 in vitro but the antiviral susceptibility of HHV-6 has not been well characterized in vivo. We retrospectively compared the HHV-6 reactivation rate in pediatric allo-SCT recipients with and without GCV prophylaxis. The HHV-6 reactivation rate at 3 weeks after allo-SCT in patients without prophylactic GCV administration was significantly higher than that in those receiving prophylactic GCV (11/28 vs 0/13, P < 0.01). Five of 36 patients without prophylactic GCV showed clinical manifestations including skin rash, interstitial pneumonitis, persistent thrombocytopenia, enterocolitis and thrombotic microangiopathy, respectively. HHV-6-associated symptoms were observed in one of the 13 patients receiving prophylactic GCV. This patient showed fever, diarrhea and graft rejection concomitantly with a sudden increase of HHV-6 DNA copy number. Patients who received GCV for treatment of HHV-6 infection showed an improvement in symptoms and/or decrease of HHV-6 copy number. Thus, GCV is effective for treating HHV-6 disease after allo-SCT in vivo.


Sedation effects of intranasal dexmedetomidine delivered as sprays versus drops on pediatric response to venous cannulation.
Mucosal atomization device (MAD) was designed to increase the bioavailability of intranasal medications by facilitating absorption, the present study aimed to evaluate and compare the sedation effects of intranasal dexmedetomidine delivered as drops versus sprays on pediatric responses to intravenous cannulation. One hundred and six pediatric patients (aged from 2 to 5years) scheduled for elective ophthalmic surgery were intranasally received a dose of 2μg/kg in 20μl/kg of dexmedetomidine for sedation to reduce response to venous cannulation. The patients were randomized into syringe group and MAD group in which dexmedetomidine was delivered as drops or sprays via syringe or MAD respectively. The primary outcome was the response to peripheral vein cannulation assessed by the FLACC scores (faces, legs, activity, cry and consolability) 30min after intranasal administration of dexmedetomidine. The secondary outcomes included acceptance for intranasal medication, sedation onset time, and needle insertion times and any adverse event at the preoperative holding area. The FLACC scores in MAD group were significantly decreased than that treated by drops (P=0.021). The acceptance for intranasal administration between both groups was comparable (P>0.05), the onset time and the incidences in two and more times of needle insertion did not differ significantly between syringe and MAD groups (all P>0.05). None of patients were required to clinically intervene in heart rates reduction and none suffered respiratory depression after administrations of dexmedetomidine in either group. Intranasal dexmedetomidine by sprays offers better sedation effects to reduce responses to venous cannulation than drops.


[A case of bilateral synchronous renal cell carcinoma managed with radical nephrectomy and ex-vivo tumor enucleation].
A 61-year-old female presented with gross hematuria and left flank pain. The arteriogram and CT scan revealed a large left renal tumor and multiple small right renal metastatic tumors. A chest X-ray and Ga scintigram showed no other metastatic lesions. A left radical nephrectomy and a right renal bench surgery were performed at the same time. The right multiple tumors were enucleated and the right kidney was autotransplanted into the right iliac fossa. The postoperative course was uneventful and the patient was free of tumor with good renal function. Renal preservation surgery was considered more beneficial for this patient than bilateral nephrectomy followed by hemodialysis.


Single-step monoclonal antibody affinity purification of human urogastrone from urine.
The use of a monoclonal antibody (MAb) affinity column to purify epidermal growth factor/urogastrone from human urine in a single-step process is described. The MAb was raised against purified recombinant human urogastrone derived from the expression of a cloned synthetic urogastrone gene. The MAb was characterised and shown to cross-react fully with recombinant and native human urogastrones. The material eluted from the column was shown to have retained biological activity. When the eluted material was run on SDS/PAGE under reducing conditions an apparently homogeneous species of 5400 Da apparent molecular weight was seen. When examined on acid PAGE, 2 molecular species corresponding to beta and gamma urogastrone were observed.


Static and fatigue failure properties of thoracic and lumbar vertebral bodies and their relation to regional density.
This study investigated (1) whether a characterization of the macroscopic architecture within the vertebral centrum would improve predictions of vertebral strength, (2) if regions in the centrum where least bone loss with age occurs are more predictive of vertebral strength, and (3) whether different patterns of the macroscopic architecture are predictive of static as compared to fatigue strength. To characterize the vertebral macroscopic architecture, a regional bone mineral density (rBMD) technique was used that estimated the cancellous density distribution (in 18 specific regions of the vertebral centrum) for vertebrae T7-L4, from spines of 20 female cadavers. Static and fatigue failure properties of whole vertebrae were obtained, and predictive models of static and fatigue failure properties of whole vertebrae were examined. We found that (1) vertebral failure properties were better predicted by combinations of vertebral regional cancellous density (multiple linear regressions) rather than by any individual region of cancellous density alone (simple linear regressions); (2) models using regions of density that demonstrated minimum decline with age [from the data of Flynn and Cody (Calcif. Tissue Int. 53, S170-S175 (1993))] resulted in better correlations with ex vivo vertebral static failure properties than models using density regions that showed maximum decline with age, and (3) static and fatigue characteristics required different density regions to reach significance. (A comparison of models predictive of static and fatigue failure properties revealed that anterior density regions were most often included in predictive models of the static properties while posterior regions were more predictive of the fatigue properties).(ABSTRACT TRUNCATED AT 250 WORDS)


What Do Reactive Fragments Actually Do in Cells?
Covalently binding molecules are frequently regarded as being generally promiscuous. In a recent study, binding selectivities and cellular target proteins of a wide variety of reactive fragments are examined in a proteome-wide context.


[The building and practice of the emergency isolation radiology information system at the department of radiology in polyclinic during the epidemic outbreak stage of SARS].
To clarify the characteristics of the hardware and software in the emergency-isolation-radiology-information-system(EIRIS) from the running experience of EIRIS during the epidemic outbreak stage of SARS(serious acute respiratory syndrom). To summarize the experience and the effects of the system, and to discuss the benefits and problems of the system. To set the dependent IP subnet address for the EIRIS. The server was in high available(HA) style. The text information was transmitted by TCP/IP protocol, and the chest X ray film information was transmitted by DICOM protocol. The client workstations were set in the SARS isolation region(at the SARS outpatient department and the SARS x-ray examination room) and the non-SARS isolation region respectively. The incidence of the SARS in medical staffs was recorded before and after the EIRIS was employed. EIRIS made the suspected and the diagnosed SARS cases away from the non-isolation area effectively. The incidence of the disease after the EIRIS running was in a downward trend compared with the incidence before the system run. EIRIS was an effective method to break the transmission route of SARS within the hospital. Network and software preparation, the training in computer of the staffs at radiology department, network maintenance and the management system are the keys to the smooth running of the EIRIS.


Input density tunes Kenyon cell sensory responses in the Drosophila mushroom body.
The ability to discriminate sensory stimuli with overlapping features is thought to arise in brain structures called expansion layers, where neurons carrying information about sensory features make combinatorial connections onto a much larger set of cells. For 50 years, expansion coding has been a prime topic of theoretical neuroscience, which seeks to explain how quantitative parameters of the expansion circuit influence sensory sensitivity, discrimination, and generalization. Here, we investigate the developmental events that produce the quantitative parameters of the arthropod expansion layer, called the mushroom body. Using Drosophila melanogaster as a model, we employ genetic and chemical tools to engineer changes to circuit development. These allow us to produce living animals with hypothesis-driven variations on natural expansion layer wiring parameters. We then test the functional and behavioral consequences. By altering the number of expansion layer neurons (Kenyon cells) and their dendritic complexity, we find that input density, but not cell number, tunes neuronal odor selectivity. Simple odor discrimination behavior is maintained when the Kenyon cell number is reduced and augmented by Kenyon cell number expansion. Animals with increased input density to each Kenyon cell show increased overlap in Kenyon cell odor responses and become worse at odor discrimination tasks.


In-vivo singlet oxygen threshold doses for PDT.
Dosimetry of singlet oxygen (1O2) is of particular interest because it is the major cytotoxic agent causing biological effects for type-II photosensitizers during photodynamic therapy (PDT). An in-vivo model to determine the singlet oxygen threshold dose, [1O2]rx,sh, for PDT was developed. An in-vivo radiation-induced fibrosarcoma (RIF) tumor mouse model was used to correlate the radius of necrosis to the calculation based on explicit PDT dosimetry of light fluence distribution, tissue optical properties, and photosensitizer concentrations. Inputs to the model include five photosensitizer-specific photochemical parameters along with [1O2]rx,sh. Photosensitizer-specific model parameters were determined for benzoporphyrin derivative monoacid ring A (BPD) and compared with two other type-II photosensitizers, Photofrin® and m-tetrahydroxyphenylchlorin (mTHPC) from the literature. The mean values (standard deviation) of the in-vivo [1O2]rx,sh are approximately 0.56 (0.26) and 0.72 (0.21) mM (or 3.6×107 and 4.6×107 singlet oxygen per cell to reduce the cell survival to 1/e) for Photofrin® and BPD, respectively, assuming that the fraction of generated singlet oxygen that interacts with the cell is 1. While the values for the photochemical parameters (ξ, σ, g, β) used for BPD were preliminary and may need further refinement, there is reasonable confidence for the values of the singlet oxygen threshold doses. In comparison, the [1O2]rx,sh value derived from in-vivo mouse study was reported to be 0.4 mM for mTHPC-PDT. However, the singlet oxygen required per cell is reported to be 9×108 per cell per 1/e fractional kill in an in-vitro mTHPC-PDT study on a rat prostate cancer cell line (MLL cells) and is reported to be 7.9 mM for a multicell in-vitro EMT6/Ro spheroid model for mTHPC-PDT. A theoretical analysis is provided to relate the number of in-vitro singlet oxygen required per cell to reach cell killing of 1/e to in-vivo singlet oxygen threshold dose (in mM). The sensitivity of threshold singlet oxygen dose for our experiment is examined. The possible influence of vascular vs. apoptotic cell killing mechanisms on the singlet oxygen threshold dose is discussed by comparing [1O2]rx,sh for BPD with 3 hr and 15 min drug-light-intervals, with the later being known to have a dominantly vascular effect. The experimental results of threshold singlet oxygen concentration in an in-vivo RIF tumor model for Photofrin®, BPD, and mTHPC are about 20 times smaller than those observed in vitro. These results are consistent with knowledge that factors other than singlet oxygen-mediated tumor cell killing can contribute to PDT damage in-vivo.


Myoglobin function evaluated in working heart tissue.
Previous modeling efforts suggest that myoglobin-facilitated diffusion contributes very little to oxygen transport in a Krebs-Henseleit perfused isolated heart when literature-derived values for the diffusion coefficient of myoglobin (DMb) were used. Simulations have strongly suggested that the contractile mechanisms of muscle may augment diffusion in working preparations. Myoglobin-facilitated oxygen diffusion may be augmented by the same mechanism. In this study, parameters of an oxygen transport to tissue model are optimized to investigate the possibility of elevated diffusion coefficients for oxygen and myoglobin in working heart tissue. The Radially-Averaged, Axially-Distributed (RAAD) model considers axial diffusion of oxygen in tissue, myoglobin facilitation of oxygen transport, and pO2-dependent oxygen consumption (Michaelis-Menten kinetics). Models are solved numerically using a variable-mesh finite-difference scheme. Parameters are optimized using a Nelder-Mead simplex routine and are chosen to minimize the sum-of-squares error (SSE) between model oxygen partial pressure (pO2) predictions and experimental pO2 data. Models are solved both with and without myoglobin facilitation. Myoglobin was found to have little effect on the oxygen distribution predicted by the models. Optimized values for the oxygen diffusion coefficient remained elevated. However, optimized values for myoglobin diffusion coefficient were found to be less than measured values. The RAAD model optimization results suggest that myoglobin does not significantly facilitate oxygen diffusion to tissue in the steady state and that myoglobin diffusion is not elevated in working heart.


Effects of acetaldehyde on the automaticity of the guinea pig sinus node.
The objective of this investigation was to characterize the effects of acetaldehyde (ACA) on sinus node automaticity (SNA). Guinea pig sinoatrial preparations superfused with Tyrode's solution at 37 degrees C were used. Intracellular microelectrodes were used to monitor SN rate (SNR). Acetaldehyde 3 X 10(-5) M had no effect on SNR, while 3 X 10(-3) M had a positive chronotropic action. The increase in SNR was associated with an increase in the slope of the slow diastolic depolarization (SDD) of subsidiary pacemaker fibers, with no change in the maximum diastolic potential (MDP). Acetaldehyde 3 X 10(-2) M exerted a biphasic effect: the SNR was enhanced and then depressed. Propranolol blocked the positive component of this chronotropic action. The negative component was not modified by propranolol, phentolamine, or atropine. It is concluded that ACA exerts both positive and negative chronotropic actions on the guinea pig sinus node. The positive component of this biphasic effect is mediated through a beta-adrenergic mechanism and it is associated with an increase in the SDD. The negative component is not due to alpha- or beta-adrenergic or muscarinic stimulation.


Presymptomatic and early pathological features of MAPT-associated frontotemporal lobar degeneration.
Early pathological features of frontotemporal lobar degeneration (FTLD) due to MAPT pathogenic variants (FTLD-MAPT) are understudied, since early-stage tissue is rarely available. Here, we report unique pathological data from three presymptomatic/early-stage MAPT variant carriers (FTLD Clinical Dementia Rating [FTLD-CDR] = 0-1). We examined neuronal degeneration semi-quantitatively and digitally quantified tau burden in 18 grey matter (9 cortical, 9 subcortical) and 13 white matter (9 cortical, 4 subcortical) regions. We compared presymptomatic/early-stage pathology to an intermediate/end-stage cohort (FTLD-CDR = 2-3) with the same variants (2 L315R, 10 P301L, 6 G272V), and developed a clinicopathological staging model for P301L and G272V variants. The 68-year-old presymptomatic L315R carrier (FTLD-CDR = 0) had limited tau burden morphologically similar to L315R end-stage carriers in middle frontal, antero-inferior temporal, amygdala, (para-)hippocampus and striatum, along with age-related Alzheimer's disease neuropathological change. The 59-year-old prodromal P301L carrier (FTLD-CDR = 0.5) had highest tau burden in anterior cingulate, anterior temporal, middle/superior frontal, and fronto-insular cortex, and amygdala. The 45-year-old early-stage G272V carrier (FTLD-CDR = 1) had highest tau burden in superior frontal and anterior cingulate cortex, subiculum and CA1. The severity and distribution of tau burden showed some regional variability between variants at presymptomatic/early-stage, while neuronal degeneration, mild-to-moderate, was similarly distributed in frontotemporal regions. Early-stage tau burden and neuronal degeneration were both less severe than in intermediate-/end-stage cases. In a subset of regions (10 GM, 8 WM) used for clinicopathological staging, clinical severity correlated strongly with neuronal degeneration (rho = 0.72, p < 0.001), less strongly with GM tau burden (rho = 0.57, p = 0.006), and did not with WM tau burden (p = 0.9). Clinicopathological staging showed variant-specific patterns of early tau pathology and progression across stages. These unique data demonstrate that tau pathology and neuronal degeneration are present already at the presymptomatic/early-stage of FTLD-MAPT, though less severely compared to intermediate/end-stage disease. Moreover, early pathological patterns, especially of tau burden, differ partly between specific MAPT variants.


[The pain referent: place in the team and leadership].
The pain referent works in close proximity to patients, and holds this specific position in addition to his or her core job. As part of an overall policy to improve pain management within healthcare establishments, their main mission is to help harmonize the practices and knowledge of healthcare professionals. They can carry out a wide range of activities, and there is currently considerable variability in the way they do so. It is essential to provide them with support through training, links with the institution via the pain control committee in particular, and local management, but also to recognize their involvement.


Oligomeric proanthocyanidins protect against HK-2 cell injury induced by oxalate and calcium oxalate monohydrate crystals.
The purpose of the study was to test whether the antioxidants oligomeric proanthocyanidins (OPCs) could provide protection against oxalate and calcium oxalate monohydrate crystals (COM) toxicity in HK-2 cells. Four groups were chosen for the study: negative control group, positive control group (COM + oxalate), OPCs group (OPCs + COM + oxalate), Vit E group (Vit E + COM + oxalate). HK-2 cells were exposed for 4, 8, 12 and 24 h. The activity of HK-2 cell was assessed by MTT. Cellular injury was assessed by activity of Na(+)/K(+) ATP enzyme. Peroxidation level was assessed by malondialdehyde (MDA) content in medium and activity of superoxide dismutase (SOD). Morphological changes of HK-2 cell after exposed for 4 and 12 h in each group were observed under Transmission electron microscope (TEM). The effects of OPCs and VitE on oxalate- and COM-exposed cells were tested. After exposed to oxalate and COM crystals, activity of cells, Na(+)/K(+) ATP enzyme and SOD enzyme showed a significant reduction, and MDA content in medium was significantly increased. OPCs group: the addition of OPCs significantly increased activity of cell, SOD and Na(+)/K(+) ATP enzyme while MDA content was significantly decreased compared with the positive control group. VitE group: compared with the positive control group, activity of HK-2 cell, Na(+)/K(+) ATP enzyme was not significantly changed while SOD activity was restored, and MDA content was significantly decreased after the addition of Vit E. Morphological structure of HK-2 cell was extremely changed as observed under TEM after exposure to high level of COM crystals and oxalate. After the addition of OPCs or Vit E, amounts of cells with vacuoles formed in cytoplasms, karyotheca dissolved and nucleolus disappeared were less than in positive control group. The morphological structure changing in OPCs group was slighter than that in Vit E group. OPCs and vitamin E administration may prevent oxalate- and COM-mediated peroxidative injury, restoring intracellular antioxidant enzyme activity. The protection rendered by OPCs was greater than that of vitamin E.


Promoter methylation patterns in Richter syndrome affect stem-cell maintenance and cell cycle regulation and differ from de novo diffuse large B-cell lymphoma.
In a fraction of patients, chronic lymphocytic leukaemia (CLL) can transform to Richter syndrome (RS), usually a diffuse large B-cell lymphoma (DLBCL). We studied genome-wide promoter DNA methylation in RS and clonally related CLL-phases of transformed patients, alongside de novo DLBCL (of non-germinal centre B type), untransformed-CLL and normal B-cells. The greatest differences in global DNA methylation levels were observed between RS and DLBCL, indicating that these two diseases, although histologically similar, are epigenetically distinct. RS was more highly methylated for genes involved in cell cycle regulation. When RS was compared to the preceding CLL-phase and with untransformed-CLL, RS presented a higher degree of methylation for genes possessing the H3K27me3 mark and PRC2 targets, as well as for gene targets of TP53 and RB1. Comparison of the methylation levels of individual genes revealed that OSM, a stem cell regulatory gene, exhibited significantly higher methylation levels in RS compared to CLL-phases. Its transcriptional repression by DNA methylation was confirmed by 5-aza-2'deoxycytidine treatment of DLBCL cells, determining an increased OSM expression. Our results showed that methylation patterns in RS are largely different from de novo DLBCL. Stem cell-related genes and cell cycle regulation genes are targets of DNA methylation in RS.