Tumor-derived biomimetic nanozyme with immune evasion ability for synergistically enhanced low dose radiotherapy.
High doses of radiation can cause serious side effects and efficient radiosensitizers are urgently needed. To overcome this problem, we developed a biomimetic nanozyme system (CF) by coating pyrite (FeS2) into tumor-derived exosomes for enhanced low-dose radiotherapy (RT). CF system give FeS2 with immune escape and homologous targeting abilities. After administration, CF with both glutathione oxidase (GSH-OXD) and peroxidase (POD) activities can significantly lower the content of GSH in tumor tissues and catalyze intracellular hydrogen peroxide (H2O2) to produce a large amount of ·OH for intracellular redox homeostasis disruption and mitochondria destruction, thus reducing RT resistance. Experiments in vivo and in vitro showed that combining CF with RT (2 Gy) can provide a substantial suppression of tumor proliferation. This is the first attempt to use exosomes bionic FeS2 nanozyme for realizing low-dose RT, which broaden the prospects of nanozymes.


[XYY syndrome: report of three cases].
Three cases of XYY syndrome (YY syndrome) were experienced. Two cases had such rare associated anomalies as hypospadias and cryptorchism. Pathogenesis, incidence, clinical symptoms, gonadal function and fertility of the patients with XYY syndrome are briefly discussed.


Identification of circular RNAs and corresponding regulatory networks reveals potential roles in the brains of honey bee workers exposed to dinotefuran.
Honey bees are important and highly efficient pollinators of agricultural crops and have been negatively affected by insecticides in recent years. Circular RNA (circRNA) plays an important role in the regulation of multiple biological and pathological processes; however, its role in the honey bee brain after exposure to dinotefuran is not well understood. Here, the expression profiles and potential modulation networks of circRNAs in the brains of workers (Apis mellifera) were comprehensively investigated using RNA sequencing and bioinformatics. In total, 33, 144, and 211 differentially expressed (DE) circRNAs were identified on the 1st, 5th and 10th days after exposure to dinotefuran, respectively. Enrichment analyses revealed that the host genes of DE circRNAs were enriched in the Hippo signaling pathway-fly, Wnt signaling pathway, and neuroactive ligand-receptor interaction. circ_0002266, circ_0005080, circ_0010239 and circ_0005415 were found to have translational potential due to the presence of an internal ribosome entry site (IRES). An integrated analysis of the DE circRNA-miRNA-mRNA networks suggest that circ_0008898 and circ_0001829 may participate in the immune response to dinotefuran exposure by acting as miRNA sponges. Our results provide invaluable basic data on A. mellifera brain circRNA patterns and a molecular basis for further study of the biological function of circRNAs in the development and immune response of honey bees.


Immune cells from patients with psoriasis are defective in inducing indoleamine 2,3-dioxygenase expression in response to inflammatory stimuli.
Indoleamine 2,3-dioxygenase (IDO) is an inducible enzyme that suppresses the immune response. The role of IDO as a negative regulator of inflammatory responses has been documented in several experimental autoimmune diseases. To explore the regulation of IDO by immune cells in psoriasis and its relation with disease severity. The expression and activity of IDO were assessed by reverse-transcriptase polymerase chain reaction, flow cytometry and high-performance liquid chromatography in peripheral blood of patients with moderate-to-severe plaque-type psoriasis. The ability of immune cells to express IDO in response to inflammatory stimuli was studied. The functional role of IDO expression was evaluated in a regulatory T cell (Treg) differentiation assay, using cocultures of immature monocyte-derived dendritic cells with autologous peripheral CD4+ T cells. Analysis of the kynurenine-to-tryptophan ratio in serum samples indicated higher IDO activity in patients with psoriasis than in healthy controls. However, correlation studies showed lower IDO activity in those patients with higher Psoriasis Area and Severity Index (PASI). Although myeloid dendritic cells from patients with psoriasis expressed higher levels of IDO than those from healthy controls, these cells did not upregulate IDO in response to a combination of tumour necrosis factor-α, interleukin (IL)-1β and IL-6 cytokines. The defective expression of IDO correlated with PASI. Immature monocyte-derived dendritic cells from patients with psoriasis also expressed low levels of IDO and induced CD4+ Treg differentiation poorly. Immune cells from patients with psoriasis have a defect in upregulating IDO in response to inflammation associated with the severity of psoriasis.


A survey of the social and psychological effects of psoriasis.
One hundred and four consecutive patients with psoriasis who were being treated in two Dublin dermatology hospital units answered a questionnaire. Social and emotional morbidity was present for many patients despite access to modern treatment facilities. Fifty five percent of patients had never experienced a complete remission from their psoriasis. A large percentage of patients avoided common social activities, for example swimming and sports; 50% felt that psoriasis had inhibited their sexual relationships and 11% of patients said they would avoid having children in case their offspring should develop the condition. Thirty-six per cent felt the hospital doctor spent most of the consultation time writing prescriptions and 59% of patients had never been given an explanatory pamphlet or book about psoriasis. To help patients to come to terms with what is for many a lifelong condition, greater efforts should be made to improve the communication at consultation, and provide more patient education.


Expression of tumor necrosis factor-α induced protein 8 like-2 contributes to the immunosuppressive property of CD4(+)CD25(+) regulatory T cells in mice.
Tumor necrosis factor-α induced protein 8 like-2 (TNFAIP8L2, TIPE2), a lately discovered negative regulator of innate immunity and cellular immunity, shares considerable sequence homology with members of the tumor necrosis factor-α induced protein 8 (TNFAIP8) family. It is preferentially expressed in lymphoid-derived and marrow-derived cells. However, it is unclear whether TIPE2 is expressed in the regulatory T cells (Tregs) to contribute to its negative regulatory property in immune response. The present study was designed to examine whether naturally occurring CD4(+)CD25(+) Tregs isolated from murine spleens expressed TIPE2 by the use of Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Based on the expression of cell surface molecules, including cytotoxic T-lymphocyte-associated antigen (CTLA)-4 and forkhead/winged helix transcription factor p3 (Foxp3) on CD4(+)CD25(+) Tregs, and cytokines including interleukin (IL)-10 as well as transforming growth factor (TGF)-β were analyzed to investigate the functional role of TIPE2 in controlling suppressive activity of CD4(+)CD25(+) Tregs. Meanwhile, IL-2, the ratio of interferon (IFN)-γ/IL-4 in CD4(+)CD25(+)Treg/CD4(+)CD25(-) T cell coculture supernatant and nuclear factor of activated T cells (NF-AT) activation in T lymphocytes were determined to examine the effects of TIPE2 on the T-cell proliferation and differentiation induced by CD4(+)CD25(+) Tregs. It was found that TIPE2 was a cytoplasmic protein expressed in CD4(+)CD25(+) Tregs, and cell surface molecules as well as cytokines (IL-10, TGF-β) expressions were significantly down-regulated when TIPE2 gene silenced by siRNA. On the other hand, CD4(+)CD25(+) Tregs treated with TIPE2 knock-down promoted T-cell proliferation as well as differentiation, and markedly upregulated IL-2 expression and intranuclear NF-AT activation. The results suggested that TIPE2 appeared to be a critical immunoregulatory molecule involved in immunosuppressive function of CD4(+)CD25(+) Tregs.


Long-Term Quality of Life After Total Gastrectomy Versus Ivor Lewis Esophagectomy.
There is scarce evidence on whether a total gastrectomy or an Ivor Lewis esophagectomy is preferred for gastroesophageal junction (GEJ) cancers regarding effects on morbidity, pathology, survival and health-related quality of life (HR-QoL). The aim of this study was to investigate the difference in long-term HR-QoL in patients undergoing total gastrectomy versus Ivor Lewis esophagectomy in a tertiary referral center. Patients with a follow-up of >1 year after a total gastrectomy or an Ivor Lewis esophagectomy for GEJ/cardia carcinoma completed the EORTC QLQ-C30 and EORTC QLQ-OG25 questionnaires. 'Problems with eating,' 'reflux,' and 'nausea and vomiting' were the primary HR-QoL endpoints. The secondary endpoints were the remaining HR-QoL domains, postoperative complications and pathology results. Thirty patients after gastrectomy and 71 after esophagectomy were included. Mean age was 63 years. Median follow-up was 2 years (range 12-84 months). Patients after gastrectomy reported less 'choking when swallowing' and 'coughing' (β = - 5.952, 95% CI - 9.437 to - 2.466; β = - 13.084, 95% CI - 18.525 to - 7.643). More lymph nodes were resected in esophagectomy group (p = 0.008). No difference was found in number of positive lymph nodes, R0 resection or postoperative complications. After a follow-up of >1 year 'choking when swallowing' and 'coughing' were less common after a total gastrectomy. No differences were found in postoperative complications or radicality of surgery. Based on this study, no general preference can be given to either of the procedures for GEJ cancer. These results support shared decision making when a choice between the two treatment options is possible.


Features of sRNA biogenesis in rice revealed by genetic dissection of sRNA expression level.
We previously conducted a QTL analysis of small RNA (sRNA) abundance in flag leaves of an immortalized rice F2 (IMF2) population by aligning sRNA reads to the reference genome to quantify the expression levels of sRNAs. However, this approach missed about half of the sRNAs as only 50% of all sRNA reads could be uniquely aligned to the reference genome. Here, we quantified the expression levels of sRNAs and sRNA clusters without the use of a reference genome. QTL analysis of the expression levels of sRNAs and sRNA clusters confirmed the feasibility of this approach. sRNAs and sRNA clusters with identified QTLs were then aligned to the high-quality parental genomes of the IMF2 population to resolve the identified QTLs into local vs. distant regulation mode. We were able to detect new QTL hotspots by considering sRNAs aligned to multiple positions of the parental genomes and sRNAs unaligned to the parental genomes. We found that several local-QTL hotspots were caused by sequence variations in long inverted repeats, which probably function as precursors of sRNAs, between the two parental genomes. The expression levels of these sRNAs were significantly associated with the presence/absence of the long inverted repeats in the IMF2 population. Moreover, we found that the variations in whole-genome sRNA species composition among different IMF2s were attributed to sRNA biogenesis genes including OsDCL2b and OsRDR2. Our results highlight that genetic dissection of sRNA expression is a promising approach to disclose new components functioning in sRNA biogenesis and new mechanisms of sRNA biogenesis.


Emerging treatment strategies for acute myeloid leukemia (AML) in the elderly.
Acute myeloid leukemia (AML) is more prevalent in older adults, with an incidence in the United States of 17.6 per 100,000 for those 65 years of age, compared with an incidence of 1.8 per 100,000 for those <65 years of age. While there have been improvements in survival during the last decade for younger patients, prognosis in elderly patients is still poor; approximately 50% achieve complete responses, but many of them relapse. With increasing age, more patients are suboptimal candidates for standard induction chemotherapy due to poor performance status, pre-existing myelodysplasia, unfavorable cytogenetics, treatment-related AML, multidrug resistance protein expression, and CD34 positivity, which are often characteristic of this patient population. In addition, the presence of comorbid conditions make many treatment options less tolerable for elderly patients. Several investigators have described subgroups showing no benefit after intensive treatment approaches in recent years. However, several novel agents have been developed to treat elderly AML patients. These include new chemotherapeutic agents, such as nucleoside analogs, as well as targeted therapies like farnesyltransferase inhibitors, tyrosine kinase inhibitors, epigenetic drugs, and antibodies. On the other hand new insights into the biology of the disease lead to a better understanding of its heterogeneity. Thus, with a variety of novel substances at hand it is increasingly important to introduce a risk-adapted approach for the optimal management of patients. This review will identify subgroups not likely to benefit from intensive chemotherapy and highlight the efficacy and tolerability of new agents in the treatment of AML.


Open-field behavior of one-clip, two-kidney hypertensive rats: the influence of pre-handling.
The relation between the development of hypertension and changes in behavior was investigated. Hypertension was induced by unilateral application of a 0.20-mm clip around the renal artery. At 4 weeks, but not at 1 week, after the operation an increase in ambulation was found. Blood pressure was significantly increased at 1 (28 mm Hg) and 4 weeks (78 mm Hg) after the operation. In a second experiment, renal hypertensive rats and controls were either regularly handled (i.e., subjected to indirect tail-cuff blood pressure measurements) or left undisturbed. At 4 weeks after clip implantation, only in the nonhandled rats with renal hypertension, an increase in open-field ambulation was found. Blood pressure was markedly increased in both handled and nonhandled renal hypertensive rats.


Aminopeptidase-B in the rat testes: isolation, functional properties and cellular localization in the seminiferous tubules.
An aminopeptidase of the B-type, with an apparent M(r) 72,000 and pI = 4.9, was isolated from rat testes and characterized. The enzyme was able to remove only Arg and/or Lys residues from L-amino acid beta-naphthylamide derivatives and from the N-terminus of several peptides. No cleavage occurred in the case of Arg-Pro bonds as found in bradykinin and substance P. The enzyme was sensitive to cysteinyl reagents and to aminopeptidase inhibitors, such as bestatin, amastatin and arphamenines A and B. The aminopeptidase activity, tested with L-Arg beta-naphthylamide and with Arg0-Met-enkephalin as substrates, was inhibited by o-phenanthroline, and restored by Zn2+ suggesting its metallopeptidase character. The partial characterization of an aminopeptidase-B activity in rat brain cortex identified a protein which is biochemically and immunologically related to the testis enzyme. By immunohistochemistry, the aminopeptidase-B was found to be particularly abundant in the seminiferous tubules at late stages of spermatogenesis and was clearly detected in a restricted area of elongated spermatids. Remarkably, the enzyme was observed to concentrate massively in the residual bodies. Since this aminopeptidase-B was able in vitro to trim out N-terminal Arg and/or Lys residues from peptides mimicking processing intermediates, it is proposed that this enzyme may be involved in propeptide and proprotein processing mechanisms in the course of spermatid differentiation.


Prediction of knee pain improvement over two years for knee osteoarthritis using a dynamic nomogram based on MRI-derived radiomics: a proof-of-concept study.
To develop and validate a nomogram to detect improved knee pain in osteoarthritis (OA) by integrating magnetic resonance imaging (MRI) radiomics signature of subchondral bone and clinical characteristics. Participants were selected from the Vitamin D Effects on Osteoarthritis (VIDEO) study. The primary outcome was 20% improvement of knee pain score over 2 years in participants administrated either vitamin D or placebo. Radiomics features of subchondral bone and clinical characteristics from 216 participants were extracted and analyzed. The participants were randomly split into the training and validation cohorts at a ratio of 8:2. Least absolute shrinkage and selection operator (LASSO) regression was used to select features and generate radiomics signatures. The optimal radiomics signature and clinical indicators were fitted into a nomogram using multivariable logistic regression model. The nomogram showed favorable discrimination performance [AUCtraining, 0.79 (95% CI: 0.72-0.79), AUCvalidation, 0.83 (95% CI: 0.70-0.96)] as well as a good calibration. Additional contributing value of fusion radiomics signature to the nomogram was statistically significant (NRI, 0.23; IDI, 0.14, P < 0.001 in training cohort and NRI, 0.29; IDI, 0.18, P < 0.05 in validating cohort). Decision curve analysis confirmed the clinical usefulness of nomogram. The radiomics-based nomogram comprising the MR radiomics signature and clinical variables achieves a favorable predictive efficacy and accuracy in differentiating improvement in knee pain among OA patients. This proof-of-concept study provides a promising way to predict clinically meaningful outcomes.


Budding uninhibited by benzimidazoles 1 might be a poor prognosis biomarker promoting the progression of papillary thyroid cancer.
Papillary thyroid carcinoma (PTC) is one of the most widespread malignant tumors of the endocrine system, with a high incidence. Budding uninhibited by benzimidazoles 1 (BUB1), one of the spindle assembly checkpoint (SAC) genes, is a multitask protein kinase required for eukaryotic chromosome segregation. Although BUB1 has been explored in several types of cancer, its biological role and molecular mechanisms in PTC remain unclear. In this study, we performed an examination of four public datasets along with local PTC cohorts and discovered that BUB1 was elevated in PTC compared to non-cancer tissues. High BUB1 expression was linked with the status of BRAFV600E , RAS, and TERT after statistical analysis. Clinically, BUB1 is associated with a variety of clinicopathological features in PTC patients. Interestingly, analysis of the TCGA database showed that BUB1 was closely associated with poor prognosis of PTC and significantly correlated with PFS. As determined by regression analysis, BUB1, and T stage were independent predictors of PTC and were related to BRAFV600E and lymph node metastatic status. By RT-qPCR, BUB1 was considerably overexpressed in PTC cell lines in comparison with normal thyroid epithelial cells. We confirmed that the knockdown of BUB1 in BCPAP and TPC1 cell lines significantly inhibited cell proliferation, cloning, and migration in vitro experiments. These results imply that BUB1 may be a significant oncogenic gene that is directly associated with the prognosis of PTC and may represent a future target for therapeutic intervention.


An Interference-Free Voltammetric Method for the Detection of Sulfur Dioxide in Wine Based on a Boron-Doped Diamond Electrode and Reaction Electrochemistry.
This paper describes a new, simple, and highly selective analytical technique for the detection of sulfur dioxide in wine, as a real sample with a relatively complicated matrix. The detection of the above analyte was based on the electrogeneration of iodine from iodide on a boron-doped diamond electrode, without modifications, in the presence of 0.1 mol dm-3 HClO4 as a supporting electrolyte. The electrogenerated iodine reacted with sulfur dioxide, forming iodide ions and sulfuric acid (i.e., a Bunsen reaction). The product of this reaction, the iodide ion, diffused back to the surface of the boron-doped diamond electrode and oxidized itself again. This chemical redox cycling enhanced the voltammetric response of the boron-doped diamond electrode. The selectivity of the determination was assured using NaOH and formaldehyde during sample preparation, and a blank was also measured and taken into account. The detection limit was estimated to be 10-6-10-7 mol dm-3. However, the content of sulfur dioxide in wine is significantly higher, which can lead to more accurate and reliable results.


Loss of differentiation in intestinal metaplasia in cancerous stomachs. A comparative morphologic study.
229 stomachs resected for duodenal and gastric ulcer and carcinoma were examined with special regard to the morphological and histochemical pattern of intestinal metaplasia (IM). The results of qualitative and semiquantitative studies were analysed statistically. Whereas duodenal and gastric ulcer cases are best discriminated by the presence or absence of IM, the strongest discriminating factor between carcinoma and gastric ulcer is the content of goblet cells in metaplastic crypts. Metaplastic crypts lined exclusively with goblet cells producing sulfated acid glycoproteins could be identified in more than one third of the cancer cases. The increase in goblet cells coincides with a loss of the more differentiated cells in the metaplastic glands, such as enterocytes, APUD cells, or Paneth cells. This "enterocoli metaplasia" seems to be specific for cancer bearing mucosa and occurs more often in cancer of intestinal type; it may represent a form of a derepressive dedifferentiation. The significance of enterocoli metaplasia as a premalignant lesion remains to be elucidated.


Sequence variation of the P gene among mumps virus strains.
We have determined nucleotide sequences of a 183-nucleotide long region of the P gene of 10 mumps virus strains after gene amplification mediated by DNA polymerase catalyzed chain reaction (PCR) and have compared them with those of two strains which had been reported earlier (K. Takeuchi et al., J. Gen. Virol., 69, 2043-2049 (1988]. It was shown that mutation is generally noncumulative, i.e., most nucleotide substitutions in earlier strains do not appear in later strains. Viruses of different lineages appeared to cocirculate, but the comparison of American and Japanese strains suggested that those isolated in one country are more closely related to each other than to those isolated in the other country.


A third fibronectin type III domain in the extracellular region of the insulin receptor family.
The insulin receptor family consists of the homologous tyrosine kinase receptors, insulin receptor (IR), insulin-like growth factor 1 receptor (IGF1R) and insulin receptor-related receptor. The three-dimensional structures of the tyrosine kinase domain of the IR and the first three extracellular domains (L1, Cys-rich and L2) of the IGF1R are known. Here we present evidence that the connecting domain of the IR family is a member of the fibronectin type II (FnIII) superfamily. Structure-based alignment of FnIII domains reveals several key residues that are also conserved in the sequence of the connecting domain. The alignment of the connecting domain with FnIII domains is in good agreement with secondary structure prediction. A model of the connecting domain shows a hydrophobic core formed by the conserved residues and is consistent with previously known biochemical data. This suggests that the IR family contains three FnIII domains in tandem in the extracellular juxtamembrane region.


Visfatin stimulates proliferation of MCF-7 human breast cancer cells.
Obesity, a condition characterized by increased fat content and altered secretion of adipokines, is a risk factor for postmenopausal breast cancer. Visfatin has recently been established as a novel adipokine that is highly enriched in visceral fat. Here we report that visfatin regulated proliferation of MCF-7 human breast cancer cells. Exogenous administration of recombinant visfatin increased cell proliferation and DNA synthesis rate in MCF-7 cells. Furthermore, visfatin activated G1-S phase cell cycle progression by upregulation of cyclin D1 and cdk2 expression. Visfatin also increased the expression of matrix metalloproteinases 2, matrix metalloproteinases 9, and vascular endothelial growth factor genes, suggesting that it may function in metastasis and angiogenesis of breast cancer. Taken together, these findings suggest that visfatin plays an important role in breast cancer progression.


Effect of thromboxane synthesis inhibition in a model of membranous nephropathy.
The effect of the thromboxane synthesis inhibitor UK 38485 on glomerular filtration rate (GFR) and proteinuria was evaluated in a rat model of unilateral membranous nephropathy. Two and 24 hours following perfusion of kidneys with cationized human IgG and i.v. administration of anti-human IgG-antiserum (in situ ICGN), glomerular thromboxane B2 (TxB2) formation was significantly higher (2 hr: 448 +/- 116 pg/mg protein/min; 24 hr: 173 +/- 21 pg/mg protein/min) compared to control (C) kidneys (2 hr: 173 +/- 21 pg/mg protein/min, P less than 0.005; 24 hr: 154 +/- 17 pg/mg protein/min, P less than 0.025). Two and seven days after induction of ICGN these differences were no longer present. Pretreatment with the thromboxane synthesis inhibitor UK 38485 prevented the decrease in GFR, which occurred two hours after induction of the glomerular disease (without UK: 161 +/- 31; with UK 325 +/- 21 microliters/100 g body wt/min). This UK 38485 effect on GFR was no longer detectable at 24 hours, two days and seven days. Initiation of glomerular immune injury was followed by significant proteinuria which averaged 250 +/- 85 mg/24 hr at day two. UK 38485 treatment, which reduced TxB2 formation in isolated glomeruli by 90% did not influence proteinuria. These data demonstrate that induction of heterologous, in situ immune complex glomerulonephritis stimulates glomerular thromboxane B2 formation, an effect which partially modulates the decrease in GFR at two hours. Thromboxane, however, does not seem to play a role in the mediation of proteinuria in this animal model.


Quads or quins? Atraumatic restricted knee flexion due to accessory quadriceps bands in children.
Recent anatomical investigations have emphasized the variability in the knee extensor apparatus, with particular attention to a fifth component of the quadriceps femoris termed the tensor vastus intermedius or accessory "quinticeps femoris." Disorders of this structure have not been described in the pediatric imaging literature. To review the clinical features, pathoanatomy and imaging findings of children presenting with derangement of the accessory quadriceps femoris, with a particular emphasis on the utility of early magnetic resonance imaging (MRI) of the thigh. This is a retrospective analysis of 3 children, ages 3-10 years at presentation, who underwent imaging evaluation with subsequent surgically proven accessory quadriceps femoris muscles. Their clinical histories, including duration of symptoms, MRI findings, intervention and surgical outcomes, are reported. All patients presented with progressive unilateral restricted knee flexion and had multiple imaging studies targeting the knee before diagnosis. Diagnosis in all patients was made on MRI of the thighs, which demonstrated a fusiform low signal intensity structure with muscle-like architecture arising from the anterior or anterolateral proximal femur and blending with the common quadriceps tendon distally. All patients underwent surgical release of the anomalous band with significant functional improvement. In cases of progressive limited knee flexion without intrinsic pathology, an accessory quadriceps muscle should be considered as an extrinsic cause. Our experience demonstrated this to be readily identifiable on MRI, with symptomatic improvement following surgical release. Early recognition of this condition should prevent unnecessary intervention such as knee arthroscopy and the debilitating loss of flexion due to delayed diagnosis.


Investigation of characteristics and risk factors of sports injuries in young soccer players: a retrospective study.
The participation of children and adolescents in sports has become increasingly frequent, including soccer. This growing involvement gives rise to concerns regarding the risk of sports injuries. Therefore, the aim of the present study was to describe the musculoskeletal injuries in young soccer players. 301 male soccer players with a mean age 14.67 ± 2.08 years were randomly recruited. The Referred Condition Inquiry was used to collect information on the mechanism of injury and anatomic site affected as well as personal data on the participants. The variables were analyzed based on the degree of association using Goodman's test for contrasts between multinomial populations, with the p < 0.05. Among the 301 athletes, 24.25% reported at least one injury. With regard to height, taller individuals reported more injuries than shorter individuals (62.5% and 37.5%, respectively; p < 0.05). Injuries were more frequent among players with a training duration greater than five years (69.65%) in comparison to those who trained for a shorter duration (30.35%) (p < 0.05). The lower limbs, especially the ankle/foot and knee, were the most affected anatomic sites. Impact was the most common mechanism of injury. The young practitioners of soccer analyzed had low rates of injury. The main causal mechanism was the impact. A taller height and longer exposure to training were the main risk factors for injury among young soccer players.


Expression of bone protein mRNA at physiological fluoride concentrations in rat osteoblast culture.
Fluoride causes an increase in the amount of unmineralized osteoid. To determine whether the increase in osteoid is due to greater protein expression in the presence of fluoride, we measured the relative amount of mRNA expressed by fetal rat calvaria cells maintained in culture for either 18 or 26 days in the presence of 0, 5, 20 or 300 microM fluoride. There were no differences in the level of expression of mRNA for collagenous or non-collagenous proteins in fluoride-treated cells as compared with control cells at 18 days in culture. Expression of mRNA for osteocalcin and alpha 1-type 1 collagen was decreased at 300 microM fluoride after 26 days culture. The amount of [3H]thymidine incorporation in cells exposed to the different amounts of fluoride was measured at various time points. Fluoride did not alter the time at which rapid cell proliferation ended. These studies indicate that at physiological serum levels, fluoride does not increase expression of mRNA by osteoblasts. The relative increase in osteoid in bone may be related to other mechanisms such as altered matrix mineralization.


Description of Flamella daurica n. sp., with notes on the phylogeny of the genus Flamella and related taxa.
A new species of naked amoebae, Flamella daurica (Amoebozoa, Conosa, Variosea), from the Russian Far East is studied and described. In addition, we isolated a strain, identified as Flamella aegyptia from Lake Ontario in Canada. Its SSU rRNA gene sequence is almost identical with that of the type strain, except for the presence of an intron in the SSU sequence. The phylogenetic analysis of a variosean alignment after inclusion of a number of environmental sequences found in GenBank confirmed that the species Talaepolella tubasferens Lahr 2012 is closely related to the Flamella clade but probably represents an independent genus and includes no less than two different species. This conclusion is supported by considerable differences between Talaepolella and Flamella in morphology and life history. Talaepolella tubasferens is an evolutionary important species demonstrating the transition from compact flamellian to expanded acramoebian morphology. The genus Flamella now includes twelve named species, which makes it one of the most species-rich genera of naked lobose amoebae.


Contractile properties of developing human fetal cardiac muscle.
The contractile properties of human fetal cardiac muscle have not been previously studied. Small-scale approaches such as isolated myofibril and isolated contractile protein biomechanical assays allow study of activation and relaxation kinetics of human fetal cardiac muscle under well-controlled conditions. We have examined the contractile properties of human fetal cardiac myofibrils and myosin across gestational age 59-134 days. Human fetal cardiac myofibrils have low force and slow kinetics of activation and relaxation that increase during the time period studied, and kinetic changes may result from structural maturation and changes in protein isoform expression. Understanding the time course of human fetal cardiac muscle structure and contractile maturation can provide a framework to study development of contractile dysfunction with disease and evaluate the maturation state of cultured stem cell-derived cardiomyocytes. Little is known about the contractile properties of human fetal cardiac muscle during development. Understanding these contractile properties, and how they change throughout development, can provide valuable insight into human heart development, and provide a framework to study the early stages of cardiac diseases that develop in utero. We characterized the contractile properties of isolated human fetal cardiac myofibrils across 8-19 weeks of gestation. Mechanical measurements revealed that in early stages of gestation there is low specific force and slow rates of force development and relaxation, with increases in force and the rates of activation and relaxation as gestation progresses. The duration and slope of the initial, slow phase of relaxation, related to myosin detachment and thin filament deactivation rates, decreased with gestation age. F-actin sliding on human fetal cardiac myosin-coated surfaces slowed significantly from 108 to 130 days of gestation. Electron micrographs showed human fetal muscle myofibrils elongate and widen with age, but features such as the M-line and Z-band are apparent even as early as day 52. Protein isoform analysis revealed that β-myosin is predominantly expressed even at the earliest time point studied, but there is a progressive increase in expression of cardiac troponin I (TnI), with a concurrent decrease in slow skeletal TnI. Together, our results suggest that cardiac myofibril force production and kinetics of activation and relaxation change significantly with gestation age and are influenced by the structural maturation of the sarcomere and changes in contractile filament protein isoforms.


Hepatitis B virus infection among health care workers at an urban teaching hospital in southern Italy: a low occupational hazard?
A prevalence study of HBV serologic markers was carried out among hospital employees of ten departments of the Second School of Medicine in Naples, an urban area with a high prevalence of HBV infection. Departments and occupational categories were selected to represent a spectrum of different exposure to B virus infection. Workers in a large electronic plant in the same geographical area were screened as controls. HBsAg prevalence was 4.8% in the hospital community and 4.0% in control group. It rises to 4.3% in the Campania Region, where all screened workers live, and in some specific areas of the same region it rises to 12%. But no significant difference among seropositivities for at least 1 marker of HBV, considered to be a better indicator of occupational hazard, was found among personnel of different departments or belonging to different occupational categories. None of the occupational and non-occupational risk factors studied was found to be significantly associated with HBV infection. Two years later, an incidence study was carried out among susceptible subjects. Seropositivity for 1 marker was 2.2% among hospital workers and 2.8% in the control group. These figures are lower than the annual attack rate (5%) required for an acceptable cost-benefit ratio of vaccination against hepatitis B. Our results indicate that in a geographical area with HBV endemicity the occupational hazard for B virus infection is low in hospital workers because of the high number of the high number of immunized subjects and the contacts with infected people out of the hospital.


The cold-inducible RNA-binding protein migrates from the nucleus to cytoplasmic stress granules by a methylation-dependent mechanism and acts as a translational repressor.
The cold-inducible RNA-binding protein (CIRP) is a nuclear 18-kDa protein consisting of an amino-terminal RNA Recognition Motif (RRM) and a carboxyl-terminal domain containing several RGG motifs. First characterized for its overexpression upon cold shock, CIRP is also induced by stresses such as UV irradiation and hypoxia. Here, we investigated the expression as well as the subcellular localization of CIRP in response to other stress conditions. We demonstrate that oxidative stress leads to the migration of CIRP to stress granules (SGs) without alteration of expression. Stress granules are dynamic cytoplasmic foci at which stalled translation initiation complexes accumulate in cells subjected to environmental stress. Relocalization of CIRP into SGs also occurs upon other cytoplasmic stresses (osmotic pressure or heat shock) as well as in response to stresses of the endoplasmic reticulum. CIRP migration into SGs is independent from TIA-1 which has been previously reported to be a general mediator of SG formation, thereby suggesting the existence of multiple pathways leading to SG formation. Moreover, deletion mutants revealed that both RGG and RRM domains can independently promote CIRP migration into SGs. However, the methylation of arginine residues in the RGG domain is necessary for CIRP to exit the nucleus to be further recruited into SGs. By RNA-tethering experiments, we also show that CIRP down-regulates mRNA translation and that this activity is carried by the carboxyl-terminal RG-enriched domain. Altogether, our findings further reveal the diversity of mechanisms by which CIRP is regulated by environmental stresses and provide new insights into CIRP cytoplasmic function.


Adjuvant radiation therapy and survival for pure tubular breast carcinoma--experience from the SEER database.
Pure tubular carcinoma of the breast (PTCB) represents a distinct subtype of invasive ductal carcinoma (IDC) that is generally thought to be associated with better prognosis than even low-grade IDC. There has been controversy as to the role of adjuvant radiation therapy (RT) in this population. We hypothesized that adjuvant RT would demonstrate a survival improvement. We queried the Surveillance, Epidemiology and End Results database for the years 1992-2007 to identify patients with pure tubular carcinomas of the breast. Patient demographics, tumor characteristics, and surgical and RT treatments were collected. Survival analysis was performed using the Kaplan-Meier method for univariate comparisons and Cox proportional hazards modeling for multivariate comparisons, stratifying on the basis of age with a cutoff age of 65. A total of 6465 patients were identified: 3624 (56.1%) patients underwent lumpectomy with RT (LUMP+RT), 1525 (23.6%) patients underwent lumpectomy alone (LUMP), 1266 (19.6%) patients received mastectomy alone (MAST), and 50 (0.8%) patients underwent mastectomy with RT (MAST+RT). When we compared the LUMP+RT and LUMP groups directly, those receiving adjuvant RT tended to be younger and were less likely to be hormone receptor-positive. Overall survival was 95% for LUMP+RT and 90% for LUMP patients at 5 years. For those 65 or younger, the absolute overall survival benefit of LUMP+RT over LUMP was 1% at 5 years and 3% at 10 years. On stratified multivariate analysis, adjuvant RT remained a significant predictor in both age groups (P=.003 in age ≤ 65 and P=.04 in age >65 patients). Other significant unfavorable factors were older age and higher T stage (age >65 only). Since sufficiently powered large scale clinical trials are unlikely, we would recommend that adjuvant radiation be considered in PTCB patients age 65 or younger, although consideration of the small absolute survival benefit is important. Adjuvant radiation can be omitted for patients older than 65.


Tissue engineered endothelial keratoplasty in rabbit: tips and tricks.
To report a detailed surgical procedure of tissue engineered endothelial keratoplasty (TEEK) in a rabbit model and its postoperative evaluation. TEEKs were prepared 7 days before transplantation by seeding human or rabbit corneal endothelial cells on either femtosecond laser-cut ultrathin human stromal lamellae (fs-UTSL) or femtosecond laser-cut human anterior lens capsule (fs-HALC). Thirty transplantations were performed on aphakic eyes. Recombinant tissue plasminogen activator (rTPA) was used throughout the surgery. The native endothelium was removed by full-surface scraping and central descemetorhexis. The transplantation was performed as a human Descemet's membrane endothelial keratoplasty. Controls included Descemetorhexis only and transplantation of carrier alone. Postoperative follow-up was performed by slit lamp and optical coherence tomography, followed by histology. Controls remained oedematous. No fibrin occurred during surgery. All but three TEEKs adhered immediately. One/6 fs-UTSL and 9/16 fs-HALC cleared perfectly (p = 0.161). All failures could be explained by at least one of the following causes intraoperative bleeding, vitreous prolapsus, early partial detachment, postoperative irido corneal synechiea/angle closure. Presumed immune rejection was observed in three rabbits only after 4 weeks. Immunostaining with anti-human CD166 allowed to perfectly differentiate human cells from rabbit cells. In successful TEEK at 3 or 4 weeks, human cells formed a normal endothelium and started migrating outside the carrier. Though the transplantation of a TEEK in rabbits is a complex model with many causes of failure, established procedure including use of rTPA allows reliable preclinical study. In addition, we suggest that fs-HALC might be a potential carrier for TEEK.


Motif analysis of amphioxus, lamprey and invertebrate estrogen receptors: toward a better understanding of estrogen receptor evolution.
The origins of steroid-dependent regulation of the vertebrate estrogen receptor (ER) are poorly understood. We used artificial-intelligence-based software to construct 12 motifs specific to the estrogen-binding domain of ERalpha and ERbeta in land vertebrates and teleosts. We mapped these ER-specific motifs onto the sequences of lamprey, amphioxus, and invertebrate and selected vertebrate ERs and estrogen-related receptors (ERR). Lamprey ER contains 11 motifs common to ERs in the training set. In contrast, amphioxus ER contains only six motifs. Unexpectedly, human and amphioxus ERRs contain nine motifs. We mapped the 12 motifs onto an alignment of human, lamprey and amphioxus ERs, which depicted residues in human ERalpha that interact with estradiol, which revealed significant differences between amphioxus ER and vertebrate ERs in the steroid-binding domain. This suggests unusual ligand recognition in amphioxus ER.


Comparison and predictors of implantable cardioverter-defibrillator therapy for primary and secondary prevention.
Implantable cardioverter-defibrillators (ICDs) are effective in detecting and treating ventricular arrhythmias. Studies on ICD therapy for different indications (primary and secondary prevention) and possible predictors of ICD therapy are limited. In this study, the incidence and type of ICD therapy were related to the indication and the underlying cardiac pathology. A single-centre, retrospective and observational study was performed of 482 patients who underwent ICD implantation for primary (53.3%) or secondary prevention (46.7%) between 2015 and 2020 at the Radboud University Medical Centre. During a median follow-up of 2.4 years (interquartile range 0.2-3.9), the occurrence of appropriate ICD therapy for primary versus secondary prevention was 9.7% and 27.6%, respectively (p < 0.001). Time to appropriate ICD therapy was significantly shorter in the secondary prevention group (p < 0.001). No difference in ICD therapy was seen for different underlying aetiologies. In the majority of cases (70%) ICD therapy was given for ventricular tachycardia (VT). The occurrence of adverse events (16.3% vs 17.3%, p = 0.772), hospitalisation for cardiovascular reasons (29.2% vs 35.1%, p = 0.559) and all-cause mortality (12.5% vs 11.6%, p = 0.763) were similar in both groups. Male gender (3.53, 95% confidence interval (CI) (1.003, 12.403), p = 0.049) and secondary prevention indication (4.90, 95% CI (1.495, 16.066), p = 0.009) were predictors of appropriate ICD therapy. The risk associated with appropriate ICD therapy is higher in secondary prevention patients, who have their first therapy within a shorter time frame after device implantation. Rates of complications, hospitalisation and all-cause mortality are comparable. Future treatment options should target the prevention of ICD therapy, mainly by preventing the recurrence of VT.


Centrifugal and centripetal connections of the cat visual cortex.
In experiments on curarized cats unit responses in the dorsal lateral geniculate body to stimulation of various zones in area 17 of the visual cortex were analyzed. Of all cells tested 69% were found to respond antidromically and 8% orthodromically; in 7.6% of cells IPSPs occurred either after an initial antidromic spike or without it. The velocities of conduction of excitation along the corticopetal fibers of the optic radiation varied from 28 to 4.3 m/sec, but the three commonest groups of fibers had conduction velocities of 28-19, 14-12, and 10-9.5 m/sec. A difference between latent periods of antidromic responses of the same neurons was found to stimulation of different zones of the visual cortex; this indicates that axons of geniculo-cortical fibers split into several branches which form contacts with several neurons in area 17 of the visual cortex. The degree and possible mechanisms of cortical influences on neurons of the lateral geniculate body are discussed.


A new technique for laparoscopic resection of a submucosal tumor on the posterior wall of the gastric fundus.
Several reports have been published which describe the technique of using an Endo GIA to resect submucosal tumors on the anterior wall of the stomach. Lesions on the posterior wall, however, especially near the esophagocardiac junction (ECJ), are difficult to resect using these reported techniques. This is because the surgeon must divide the omentum and enter the omental bursa in order to use a similar extraluminal technique. Furthermore, special care must be taken to ensure that resections do not involve the ECJ and narrow the esophagus. In order to overcome these difficulties, we have proposed a new technique for the laparoscopic excision of a submucosal tumor located on the posterior wall of the gastric fundus. The principle of this procedure involves the intraluminal resection of the submucosal tumor, including the surrounding stomach wall, using the Endo GIA. This technique is safe, simple, and effective. We believe that we are the first to address the excision of a submucosal lesion by resecting the full thickness of the posterior gastric wall lesion intraluminally.


Posterior prolongation of the cartilaginous nasal septum: an under-utilised source of autologous graft material.
To assess the clinical and radiological characteristics of the posterior prolongation of the cartilaginous nasal septum, an under-utilised source of autologous cartilage for nasal reconstruction. Consecutive patients undergoing primary, external approach rhinoplasty were included. The septal cartilage was assessed intra-operatively prior to routine harvest. Cartilage use was recorded and post-operative cosmesis noted. Computed tomography scans from a separate patient group, with no septal surgery, were used to assess septal cartilage dimensions. Of the 25 rhinoplasty patients studied, 24 had harvestable septal cartilage, with a posterior prolongation mean length ± standard deviation of 24.3 ± 8.40 mm, mean height of 4.33 ± 0.34 mm and mean width of 1.1 ± 0.35 mm. The mean post-operative cosmesis score was +2.41 ± 0.71 at a mean follow up of 45 ± 8.7 weeks. All 25 radiology patients had visible posterior prolongations on computed tomography (mean length, 18.1 ± 5.1 mm; mean height, 4.2 ± 1.1 mm; mean width 1.5 ± 0.63 mm). Harvesting of the posterior prolongation would increase by 25 per cent the cartilage area available for autologous grafts. Endoscopic guidance aids this process. Cartilage is most commonly used for overlay grafts, with good cosmesis. The posterior prolongation is demonstrated on computed tomography, although dimensions may be underestimated.


The effect of hormone therapy on peripheral blood leukocytic subset distribution in stage D prostatic cancer patients.
The distribution of mononuclear cell types found in the peripheral blood of patients bearing carcinoma of the prostate were compared by stage and to a control group using monoclonal antibody techniques. Patients with lower stage disease (A, B) had no significant alteration in subset distribution when compared to a control group, while those with higher stage disease (D) had significant deviations. Stage D patients had a decreased representation of helper-inducer T cells and an increased representation of suppressor-cytotoxic T cells, with an overall reduction in the total T cell content. In addition elevated levels of monocytic, granulocytic and null cells were recognized by the polyspecific OKM1 antibody. These differences were in part reversible following hormonal therapy. Such alterations in the ratios between the various T cell populations could be useful in patient staging and treatment selection.


Oxidative stress and cell damage in a model of precancerous lesions and advanced hepatocellular carcinoma in rats.
Hepatocellular carcinoma (HCC) is the third most frequent cause of cancer deaths throughout the world. This study was aimed to analyze oxidative stress and cell damage in a multistage model of liver carcinogenesis induced by diethylnitrosamine (DEN) in rats. Male Wistar rats weighing 145-150 g were divided into three groups: control, precancerous lesions (PL) (which received 100 mg DEN once a week every 6 weeks up to 28 weeks), and advanced HCC (50 mg DEN once/twice per week up to 19 weeks). Lipid peroxidation (TBARS), superoxide dismutase (SOD) activity, and expression of transforming growth factor-1 beta (TGF)-1β, endothelial and inducible nitric oxide syntahese (eNOS, iNOS), NADPH quinone oxireductase (NQO)-1, nuclear factor erythroid 2-related factor (NrF)2, kelch-like ECH-associated protein (Keap)1 and heat shock protein (HSP)70 were measured. TBARS concentration was augmented in the PL and advanced HCC groups. SOD activity, TGF-1β and Nrf2 expression were higher in animals with precancerous lesions. In advanced HCC, expression of NQO1 and iNOS increased while there was a decrease in HPS70 expression. Data obtained provide evidence for the differential activation of proteins involved in oxidative stress and cell damage during progression of carcinogenesis in an animal model of HCC.


Return to Play and Performance After Anterior Cruciate Ligament Reconstruction in the National Women's Soccer League.
The anterior cruciate ligament (ACL) is commonly injured in elite-level female athletes, which usually requires ACL reconstruction (ACLR). To analyze return to play (RTP) and changes in performance of players in the National Women's Soccer League (NWSL) after ACLR. Descriptive epidemiology study. NWSL players who sustained an ACL tear and underwent surgery between the 2013 and 2020 seasons were identified by multiple online resources. Players were classified as forwards, defenders, midfielders, and goalkeepers. RTP was assessed according to games played, games started, percentage of minutes played, plus/minus net per 90 minutes (a measure of a player's contribution to their team's performance while on the field), goals scored, and assists. A subanalysis was performed based on the median age at the time of the injury (≤24 vs ≥25 years). Nonparametric testing methods were used throughout the analysis. A total of 30 NWSL athletes were included. Midfielders had the highest percentage of injuries (n = 11; 36.7%), followed by forwards (n = 10; 33.3%). Overall, 27 players returned to the NWSL at a median of 12.1 months (IQR, 10.9-14.3 months), constituting a 90.0% RTP rate. There was a significant decrease in the percentage of minutes played from 1 year before the injury to 1 year after the injury (median, 87.9% [IQR, 80.7%-90.6%] vs 25.1% [IQR, 16.3%-57.2%], respectively; P = .031). Forwards and midfielders had a significant decrease in the number of assists from 1 year before the injury to 1 year after the injury (median, 3.0 [IQR, 1.0-3.0] vs 0.0 [IQR, 0.0-1.0], respectively; P = .037) as well as the number of goals scored when averaging across 2 seasons before the injury to 2 seasons after the injury (median, 3.0 [IQR, 1.5-5.5] vs 1.0 [IQR, 0.5-3.5], respectively; P = .031). On subanalysis, older players started in significantly more games (median, 12.0 [IQR, 3.8-18.5] vs 3.0 [IQR, 0.5-6.0], respectively; P = .048) and had a higher percentage of minutes played (median, 63.0% [IQR, 18.8%-77.3%] vs 14.9% [IQR, 2.0%-21.2%], respectively; P = .046) at 1 year after the injury versus younger players. There was a 90.0% RTP rate after ACLR in the NWSL. Players who returned to the NWSL had a lower percentage of minutes played in their first year after RTP, with older players starting in more games and having a greater percentage of minutes played. Compared with preinjury performance, forwards and midfielders had a significant decrease in the number of assists at 1 year after the injury as well as the number of goals scored at 2 years after the injury.


Reduced haloperidol: effects on striatal dopamine metabolism and conversion to haloperidol in the rat.
Acute injection of rats with either haloperidol or reduced haloperidol (1 mg/kg, IP) greatly increased the striatal concentrations of the acidic dopamine metabolites, indicating enhanced turnover of dopamine. The effect of reduced haloperidol was almost as great as that of haloperidol. Reduced haloperidol, however, was much less efficient (about 400 times) than haloperidol in displacing [3H]spiperone binding to striatal membranes in vitro. In agreement with the above results, reduced haloperidol was found to be oxidized to haloperidol, so that 2 h after injection of reduced haloperidol the concentrations of haloperidol and reduced haloperidol were equal in the striatum. The apparent conversion of reduced haloperidol to haloperidol was much quicker in liver than in plasma or brain, and it is suggested that the conversion primarily occurs in the liver. Before drawing any definite conclusion about the possible central activity of reduced haloperidol, further studies with other animal species are needed.


Desire to have children and preferences regarding to pre-pregnancy counselling in women with SLE.
Pre-pregnancy counselling in women with systemic lupus erythematosus (SLE) is important in order to improve knowledge on the risks of pregnancy and to optimize pregnancy outcomes. Knowledge on the preferences of women with SLE regarding pre-pregnancy counselling have not yet been studied. In a closely monitored cohort of women with SLE we enquired about the present status of their wish to have children, and wish for and experiences with pre-pregnancy counselling. A questionnaire developed by physicians in collaboration with two women with SLE was sent to all (n = 177) women participating in the Amsterdam SLE cohort. The questionnaire comprised 32 items, of which 15 focused on the above-mentioned three themes. A total of 124 women (70%) returned the questionnaire. The median disease duration was 13 years (interquartile range 9-19). Childlessness occurred in 51 women and 31% declared this was due to SLE [conscious decision (21%), stringent medical advice (6%), infertility due to medication (4%)]. Half of the women preferred the first pre-pregnancy counselling immediately after the SLE diagnosis (53%), together with their partner (69%). Information given by healthcare providers (81%) was preferred over information provided via brochures (35%) or the internet (26%). Pre-pregnancy face-to-face counselling from a rheumatologist and/or gynaecologist separately was preferred in 54%. One-third of women attributed their childlessness to SLE-related reasons. Pre-pregnancy counselling was preferred shortly after the onset of the disease in a non-multidisciplinary setting. The results of this study underline the importance of timely pre-conceptional counselling by healthcare providers on fertility, risks and pregnancy outcomes in women with SLE.


Gastrointestinal bleeding after operation for pancreatic cancer.
Gastrointestinal bleeding after surgery for cancer of the pancreas contributes significantly to patient morbidity and to patient mortality, if the bypass was performed for palliation. Bleeding after resection of the pancreatic tumor appears to be amenable to therapy, and unaltered by the addition of vagotomy at the time of surgery. Patients undergoing palliative surgery who are expected to live beyond the postoperative period may well benefit from measures to reduce the risk of gastrointestinal bleeding.


Genome editing of an African elite rice variety confers resistance against endemic and emerging Xanthomonas oryzae pv. oryzae strains.
Bacterial leaf blight (BB) of rice, caused by Xanthomonas oryzae pv. oryzae (Xoo), threatens global food security and the livelihood of small-scale rice producers. Analyses of Xoo collections from Asia, Africa and the Americas demonstrated complete continental segregation, despite robust global rice trade. Here, we report unprecedented BB outbreaks in Tanzania. The causative strains, unlike endemic African Xoo, carry Asian-type TAL effectors targeting the sucrose transporter SWEET11a and iTALes suppressing Xa1. Phylogenomics clustered these strains with Xoo from Southern-China. African rice varieties do not carry effective resistance. To protect African rice production against this emerging threat, we developed a hybrid CRISPR-Cas9/Cpf1 system to edit all known TALe-binding elements in three SWEET promoters of the East African elite variety Komboka. The edited lines show broad-spectrum resistance against Asian and African strains of Xoo, including strains recently discovered in Tanzania. The strategy could help to protect global rice crops from BB pandemics.


Quantitation of phospholipids on thin layer chromatographic plates using a desk-top scanner.
A method for quantitating phospholipids separated on thin layer chromatographic plates by computer-assisted photodensitometry is described. After development, the plates are stained with molibdic reagent and the image obtained is acquired as TIFF file in the memory of a personal computer. The color intensity of the single spots of the digitalized image is analyzed using a dedicated software. Sensitivity and reproducibility are adequate for most of the needs of lipid chemist. When compared to conventional photodensitometric procedures, the present method offers the advantage of requiring a much cheaper hardware.


Multimodal imaging shows fibrosis architecture and action potential dispersion are predictors of arrhythmic risk in spontaneous hypertensive rats.
Hypertensive heart disease (HHD) increases risk of ventricular tachycardia (VT) and ventricular fibrillation (VF). The roles of structural vs. electrophysiological remodelling and age vs. disease progression are not fully understood. This cross-sectional study of cardiac alterations through HHD investigates mechanistic contributions to VT/VF risk. Risk was electrically assessed in Langendorff-perfused, spontaneously hypertensive rat hearts at 6, 12 and 18 months, and paced optical membrane voltage maps were acquired from the left ventricular (LV) free wall epicardium. Distributions of LV patchy fibrosis and 3D cellular architecture in representative anterior LV mid-wall regions were quantified from macroscopic and microscopic fluorescence images of optically cleared tissue. Imaging showed increased fibrosis from 6 months, particularly in the inner LV free wall. Myocyte cross-section increased at 12 months, while inter-myocyte connections reduced markedly with fibrosis. Conduction velocity decreased from 12 months, especially transverse to the myofibre direction, with rate-dependent anisotropy at 12 and 18 months, but not earlier. Action potential duration (APD) increased when clustered by age, as did APD dispersion at 12 and 18 months. Among 10 structural, functional and age variables, the most reliably linked were VT/VF risk, general LV fibrosis, a measure quantifying patchy fibrosis, and non-age clustered APD dispersion. VT/VF risk related to a quantified measure of patchy fibrosis, but age did not factor strongly. The findings are consistent with the notion that VT/VF risk is associated with rate-dependent repolarization heterogeneity caused by structural remodelling and reduced lateral electrical coupling between LV myocytes, providing a substrate for heterogeneous intramural activation as HHD progresses. KEY POINTS: There is heightened arrhythmic risk with progression of hypertensive heart disease. Risk is related to increasing left ventricular fibrosis, but the nature of this relationship has not been quantified. This study is a novel systematic characterization of changes in active electrical properties and fibrotic remodelling during progression of hypertensive heart disease in a well-established animal disease model. Arrhythmic risk is predicted by several left ventricular measures, in particular fibrosis quantity and structure, and epicardial action potential duration dispersion. Age alone is not a good predictor of risk. An improved understanding of links between arrhythmic risk and fibrotic architectures in progressive hypertensive heart disease aids better interpretation of late gadolinium-enhanced cardiac magnetic resonance imaging and electrical mapping signals.


Correlated light and electron microscopic imaging of multiple endogenous proteins using Quantum dots.
The importance of locating proteins in their context within cells has been heightened recently by the accomplishments in molecular structure and systems biology. Although light microscopy (LM) has been extensively used for mapping protein localization, many studies require the additional resolution of the electron microscope. Here we report the application of small nanocrystals (Quantum dots; QDs) to specifically and efficiently label multiple distinct endogenous proteins. QDs are both fluorescent and electron dense, facilitating their use for correlated microscopic analysis. Furthermore, QDs can be discriminated optically by their emission wavelength and physically by size, making them invaluable for multilabeling analysis. We developed pre-embedding labeling criteria using QDs that allows optimization at the light level, before continuing with electron microscopy (EM). We provide examples of double and triple immunolabeling using light, electron and correlated microscopy in rat cells and mouse tissue. We conclude that QDs aid precise high-throughput determination of protein distribution.


The behavior of heavy metal release from sulfide waste rock under microbial action and different environmental factors.
The dissolution of heavy metals from the waste rock is controlled by many factors. Herein, we investigated the release behavior of iron (Fe), chromium (Cr), copper (Cu), and zinc (Zn) from sulfide waste rock under the actions of microorganisms and different environmental factors (solution pH value, particle size of waste rock, temperature, Fe3+ concentration). The release quantity of heavy metals was negatively correlated with pH and particle size and positively correlated with ambient temperature and Fe3+ concentration. Under the experimental conditions of pH value of 3.0, temperature of 35°C, and waste stone particle size of less than 0.075 mm,, the release quantity of Fe, Cr, Cu, and Zn reached 3680, 18.32, 132.20, 26.60 mg·kg-1 after 20 days of leaching, respectively. Rising the temperature to 45 °C, Fe, Cr, Cu, and Zn release quantities increased to 89.30, 5.81, 105.08, and 28.00 mg·kg-1. Six hundred milligrams per liter Fe3+ increased the release of heavy metals considerably (2.63-65.48 folds). The presence of microorganisms can significantly facilitate the release of heavy metals. Compared to the control group, the release quantities of Fe, Cr, Cu, and Zn increased 4.29, 3.17, 1.54, and 2.39 times, respectively. In addition, the waste rock under microbial action was more seriously corroded than that under chemical factors. The release behavior of these four heavy metals was consistent with the interfacial chemical reaction control model, indicating that the reactions mainly occurred on the surface of the waste rock. This study provides an essential reference for the study of heavy metal leaching behavior.


Diphtheria toxin and its mutant crm 197 differ in their interaction with lipids.
The interaction of diphtheria toxin and its enzymatically deficient mutants crm 176 and crm 197 with liposomes has been studied by turbidity measurement and hydrophobic photolabelling with photoactivatable phosphatidylcholines. Diphtheria toxin and crm 176 at neutral pH bind to the surface of lipid bilayers while crm 197 also appears to interact with the fatty acid chains of phospholipids. All proteins undergo a change in conformation over the same range of acidic pH and become able to insert in the lipid bilayer. The tighter lipid interaction of crm 197 may account for its higher cell association constant. The possibility is discussed that the binding of diphtheria toxin to cells is mediated by both a protein receptor and an interaction with the head group of phospholipids.


Substrate DNA length regulates the activity of TET 5-methylcytosine dioxygenases.
The ten-eleven translocation (TET) isoforms (TET1-3) play critical roles in epigenetic transcription regulation. In addition, mutations in the TET2 gene are frequently detected in patients with glioma and myeloid malignancies. TET isoforms can oxidize 5-methylcytosine to 5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxylcytosine, by iterative oxidation. The in vivo DNA demethylation activity of TET isoforms may depend on many factors including enzyme's structural features, its interaction with DNA-binding proteins, chromatin context, DNA sequence, DNA length, and configuration. The rationale for this study is to identify the preferred DNA length and configuration in the substrates of TET isoforms. We have used a highly sensitive LC-MS/MS-based method to compare the substrate preference of TET isoforms. To this end, four DNA substrate sets (S1, S2, S3, S4) of different sequences were chosen. In addition, in each set, four different lengths of DNA substrates comprising 7-, 13-, 19-, and 25-mer nucleotides were synthesized. Each DNA substrate was further used in three different configurations, that is, double stranded symmetrically-methylated, double stranded hemi-methylated, and single stranded single-methylated to evaluate their effect on TET-mediated 5mC oxidation. We demonstrate that mouse TET1 (mTET1) and human TET2 (hTET2) have highest preference for 13-mer dsDNA substrates. Increasing or decreasing the length of dsDNA substrate reduces product formation. In contrast to their dsDNA counterparts, the length of ssDNA substrates did not have a predictable effect on 5mC oxidation. Finally, we show that substrate specificity of TET isoforms correlates with their DNA binding efficiency. Our results demonstrate that mTET1 and hTET2 prefer 13-mer dsDNA as a substrate over ssDNA. These results may help elucidate novel properties of TET-mediated 5mC oxidation and help develop novel diagnostic tools to detect TET2 function in patients.


Evaluation of a combined drug-delivery system for proteins assembled with polymeric nanoparticles and porous microspheres; characterization and protein integrity studies.
This work presents an evaluation of the adsorption/infiltration process in relation to the loading of a model protein, α-amylase, into an assembled biodegradable polymeric system, free of organic solvents and made up of poly(D,L-lactide-co-glycolide) acid (PLGA). Systems were assembled in a friendly aqueous medium by adsorbing and infiltrating polymeric nanoparticles into porous microspheres. These assembled systems are able to load therapeutic amounts of the drug through adsorption of the protein onto the large surface area characteristic of polymeric nanoparticles. The subsequent infiltration of nanoparticles adsorbed with the protein into porous microspheres enabled the controlled release of the protein as a function of the amount of infiltrated nanoparticles, since the surface area available on the porous structure is saturated at different levels, thus modifying the protein release rate. Findings were confirmed by both the BET technique (N2 isotherms) and in vitro release studies. During the adsorption process, the pH of the medium plays an important role by creating an environment that favors adsorption between the surfaces of the micro- and nano-structures and the protein. Finally, assays of α-amylase activity using 2-chloro-4-nitrophenyl-α-D-maltotrioside (CNP-G3) as the substrate and the circular dichroism technique confirmed that when this new approach was used no conformational changes were observed in the protein after release.


Association of dowry practices with perceived marital life and intimate partner violence.
To understand the perceptions of women about the influence of dowry customs on their marital life and on intimate partner violence. The cross-sectional study was conducted in Karachi between 2008 to 2010, and comprised married women of reproductive age. Data was collected through a valid World Health Organisation questionnaire which was validated for the local context after translation into Urdu. Data was analysed using SPSS 10. Of the 810 women approached, 759(93.7%) formed the final sample. Of them, 447(59%) women and 307(40.4%) of the husbands were aged 25-35 years. Women in arranged marriages involving dowry transaction reported more positive marital outcomes (adjusted odds ratio: 11.5). Consenting to a marriage was positively associated with positive marital life (adjusted odds ratio: 36.8), and the same was the case when the marriage was contingent on dowry transaction (adjusted odds ratio: 10.4). Provision of a dowry, however, was not protective from physical (adjusted odds ratio: 3.7), sexual (adjusted odds ratio: 3.7) or psychological violence (adjusted odds ratio: 8.9). Dowry practices exist in Pakistani culture despite the fact that dowry wives were found to have no protection against intimate partner violence. However, women perceived that the provision of dowry to groom's family had a positive impact on marital life.


Incidence and predictors of treatment-related conjugated hyperbilirubinemia during early treatment phases for children with acute lymphoblastic leukemia.
Conjugated hyperbilirubinemia (CHB) and liver transaminase elevation are known complications of acute lymphoblastic leukemia (ALL) therapy, but host risk factors are poorly understood. Among 373 children diagnosed with ALL between 2011 and 2016, clinically significant CHB and transaminase elevation were observed in 15 (4.0%) and 12 (3.2%) children, respectively, during induction and consolidation. Body mass index ≥95th percentile (odds ratio 9.20, 95% confidence interval 2.56-32.96) was the only host factor independently associated with CHB, and no host factors were associated with transaminase elevation. Obese patients warrant closer monitoring of hepatic function to facilitate early intervention prior to the development of severe, adverse hepatic events.


Measuring Ag-specific immune responses: understanding immunopathogenesis and improving diagnostics in infectious disease, autoimmunity and cancer.
Characterization of antigen-specific immune responses at the single-cell level has been made possible by recent advancements in reagent and technology development, combined with increasing knowledge of molecular mechanisms. Fluorescently labelled MHC-peptide multimers and antigens identify directly specific T and B cells, respectively, whereas dynamic assays exploit mediator production or secretion, or the changes in surface expression of other proteins, to identify specific lymphocytes--some techniques enabling the recovery of viable cells. Meanwhile, multiparameter flow cytometry has emerged as the most versatile platform for integrating most of these methods. As the complexity of experimental data increases, so does the level of technical sophistication required for analysis and interpretation, both in terms of basic research and modern medicine, with new applications for infectious diseases, autoimmunity and cancer.


2D and 3D ordered columnar liquid crystal phases by bundles of bolaamphiphiles with swallow-tail side chains.
Two new mesophases, a longitudinally correlated and a noncorrelated (columnar) hexagonal phase, were obtained for a polyphilic bolaamphiphile with laterally attached swallow-tail chains; the unique feature of these mesophases, which makes them distinct from previously reported LC phases, is the organization of rod-like units in bundles parallel to the column axis.


All-natural environmentally degradable poly (butylene terephthalate-co-caprolactone): A theoretical and experimental study of its degradation properties and mechanisms.
The design and production of materials with excellent mechanical properties and biodegradability face significant challenges. Poly (butylene terephthalate-co-caprolactone) copolyesters (PBTCL) is obtained by modifying the engineering plastic polybutylene terephthalate (PBT) with a simple one-pot process using readily biodegradable ε-caprolactone (ε-CL). The material has mechanical properties comparable to those of commercial biodegradable copolyester PBAT. Besides, this copolyester exhibited remarkable degradability in natural environments such as soil and ocean, for example, PBTCL1.91 lost >40 % of its weight after 6 months of immersion in the Bohai Sea. The effect and diversity of specific microorganisms acting on degradation in the ocean were analyzed by 16 s rDNA gene sequencing. Theoretical calculations such as Fukui function and DFT, and experimental studies on water-soluble intermediates and residual matrixes produced after degradation, confirmed that the insertion CL units not only act as active sites themselves susceptible to hydrolysis reactions, but also promote the reactivity of ester bonds between aromatic segments. This work provides insight for the development of novel materials with high performance and environmental degradability.


Structural evolution of the P22-like phages: comparison of Sf6 and P22 procapsid and virion architectures.
Coat proteins of tailed, dsDNA phages and in herpesviruses include a conserved core similar to the bacteriophage HK97 subunit. This core is often embellished with other domains such as the telokin Ig-like domain of phage P22. Eighty-six P22-like phages and prophages with sequenced genomes share a similar set of virion assembly genes and, based on comparisons of twelve viral assembly proteins (structural and assembly/packaging chaperones), these phages are classified into three groups (P22-like, Sf6-like, and CUS-3-like). We used cryo-electron microscopy and 3D image reconstruction to determine the structures of Sf6 procapsids and virions (~7Å resolution), and the structure of the entire, asymmetric Sf6 virion (16-Å resolution). The Sf6 coat protein is similar to that of P22 yet it has differences in the telokin domain and in its overall quaternary organization. Thermal stability and agarose gel experiments show that Sf6 virions are slightly less stable than those of P22. Finally, bacterial host outer membrane proteins A and C were identified in lipid vesicles that co-purify with Sf6 particles, but are not components of the capsid.


Phenotypic correction of hemophilia A in sheep by postnatal intraperitoneal transplantation of FVIII-expressing MSC.
We recently re-established a line of sheep that accurately mimics the clinical symptoms and genetics of severe hemophilia A (HA). Here, we tested a novel, nonablative transplantation therapy in two pediatric HA animals. Paternal mesenchymal stem cells (MSC) were transduced with a porcine FVIII-encoding lentivector and transplanted via the intraperitoneal route without preconditioning. At the time of transplantation, these animals had received multiple human FVIII treatments for various spontaneous bleeds and had developed debilitating hemarthroses, which produced severe defects in posture and gait. Transplantation of transduced MSC resolved all existent hemarthroses, and spontaneous bleeds ceased. Damaged joints recovered fully; the animals regained normal posture and gait and resumed normal activity. Despite achieving factor-independence, a sharp rise in pre-existent Bethesda titers occurred following transplantation, decreasing the effectiveness and duration of therapy. Postmortem examination revealed widespread engraftment, with MSC present within the lung, liver, intestine, and thymus, but particularly within joints affected at the time of transplantation, suggesting MSC homed to sites of ongoing injury/inflammation to release FVIII, explaining the dramatic improvement in hemarthrotic joints. In summary, this novel, nonablative MSC transplantation was straightforward, safe, and converted life-threatening, debilitating HA to a moderate phenotype in a large animal model.


Optimizing resolution in multidimensional NMR by three-way decomposition.
Resolution depends on the number of points sampled in a FID; in indirectly detected dimensions it is an important determinant of the total experiment time. Based on the high redundancy present in NMR data, we propose the following timesaving scheme for three-dimensional spectra. An extensive grid of discrete t1- and t2-values is used, which increases resolution while preserving the spectral width. Total experiment time is reduced by avoiding the recording of t3-FIDs for selected pairs of t1 and t2; typically the recording is omitted for about 75% of the (t1,t2) combinations. These data sets are referred to as sparse, and post-experimental processing making optimal use of spectral redundancy provides the missing, non-recorded data. We have previously shown that three-way decomposition (TWD) within the MUNIN approach provides a practical way to process dense NMR data sets. Here, a novel TWD algorithm [Ibraghimov, (2002) Numer. Linear Algebra Appl. 9, 551-565] is used to complement a sparsely recorded time-domain data set by providing the missing FIDs for all (t1,t2) combinations omitted in the experiment. A necessary condition is that for each t1-value at least a few FIDs are recorded, and similar for each t2-value. The method is demonstrated on non-uniformly sampled (15)N-NOESY-HSQC data sets recorded for the 14 kD protein azurin. The spectra obtained by TWD, reconstruction and ordinary transform to frequency-domain are, in spite of the large number of signals and the high dynamic range typical for NOESYs, highly similar to a corresponding reference spectrum, for which all (t1,t2) combinations were recorded.


Smoking habits are an independent prognostic factor in patients with lung cancer.
The role of tobacco in the pathogenesis of lung cancer (LC) has been clearly established. Based on the epidemiological evidence that smoking may influence LC progression, we investigated the idea that smoking behavior could be associated with overall survival (OS) in this group of patients. A total of 351 patients with LC (311 men and 40 women) were reviewed. Smoking status was assessed as tobacco users or non-users. To calculate pack-years of smoking, the average of number of cigarettes smoked per day was divided by 20 to give packs per day, and then multiplied by the total number of years of smoking. OS was the main outcome measure. The mean follow-up was 3.3 ± 1.2 years. Kaplan-Meier plots of OS by use of tobacco revealed significant differences by smoking status (log-rank = 5.44, P < 0.01), indicating a reduced survival rate in tobacco users. The effect on OS of the amount of cigarette smoking was also evident when we subdivided the former and current smokers into ≤7 (mean value) pack-years and >7 pack-years groups (log-rank = 4.27, P < 0.05). After adjusting for all potential confounders, tobacco smoking retained its independent prognostic significance for OS (hazard ratio = 1.53, 95% confidence interval = 1.19-2.17, P = 0.02). Our data indicate that cigarette smoking is significantly associated with a poor prognosis among patients diagnosed with LC in a dose-dependent manner.


Corticosteroid injections reduce size of rheumatoid nodules.
Symptomatic rheumatoid nodules are frequently surgically treated. Injection with steroids might be an alternative treatment. To determine whether injection with triamcinolon acetonide reduces the size of rheumatoid nodules, we randomized twenty patients with symptomatic nodules to either triamcinolon acetonide 40 mg/ml plus lidocaine 2% or lidocaine 1% (placebo). We measured the nodules before injection and 2, 4, 8, and 12 weeks after injection. Possible side effects were recorded. We found that the volume of the nodules injected with triamcinolon acetonide reduced significantly (p = 0.011), from 130 to 8 mm(3) (median calculated size) at 12 weeks, compared with baseline. Furthermore, at 12 weeks, the difference between the groups was significant (p = 0.03). The median size of the placebo nodules diminished as well, from 358 to 237 mm(3), but this was not significant. Pain at injection was the only side effect, equally distributed in both treatment groups. Injection with triamcinolon acetonide seems to be an alternative for surgery of rheumatoid nodules. No adverse events occurred but the limited sample does not allow definitive conclusions.


The impact of 177Lu-octreotide therapy on 99mTc-MAG3 clearance is not predictive for late nephropathy.
Peptide Receptor Radionuclide Therapy (PRRT) for the treatment of neuroendocrine tumors may lead to kidney deterioration. This study aimed to evaluate the suitability of 99mTc-mercaptoacetyltriglycine (99mTc--MAG3) clearance for the early detection of PRRT-induced changes on tubular extraction (TE). TE rate (TER) was measured prior to 128 PRRT cycles (7.6±0.4 GBq 177Lu-octreotate/octreotide each) in 32 patients. TER reduction during PRRT was corrected for age-related decrease and analyzed for the potential to predict loss of glomerular filtration (GF). The GF rate (GFR) as measure for renal function was derived from serum creatinine. The mean TER was 234 ± 53 ml/min/1.73 m² before PRRT (baseline) and 221 ± 45 ml/min/1.73 m² after a median follow-up of 370 days. The age-corrected decrease (mean: -3%, range: -27% to +19%) did not reach significance (p=0.09) but significantly correlated with the baseline TER (Spearman p=-0.62, p<0.001). Patients with low baseline TER showed an improved TER after PRRT, high decreases were only observed in individuals with high baseline TER. Pre-therapeutic TER data were inferior to plasma creatinine-derived GFR estimates in predicting late nephropathy. TER assessed by 99mTc-MAG3-clearance prior to and during PRRT is not suitable as early predictor of renal injury and an increased risk for late nephropathy.


Use of protection motivation theory, affect, and barriers to understand and predict adherence to outpatient rehabilitation.
Protection motivation theory (PMT) has been used in more than 20 different health-related fields to study intentions and behavior, albeit primarily outside the area of injury rehabilitation. In order to examine and predict patient adherence behavior, this study was carried out to explore the use of PMT as a screening tool in a general sample of people with orthopedic conditions. New patients who were more than 18 years old and who were prescribed 4 to 8 weeks of physical therapy treatment (n=229) were administered a screening tool (Sports Injury Rehabilitation Beliefs Scale, Positive and Negative Affect Schedule, and a barriers checklist) prior to treatment. Participants' adherence was assessed with several attendance measures and an in-clinic assessment of behavior. Statistical analyses included correlation, chi-square, multiple regression, and discriminant function analyses. A variety of relationships among affect, barriers, and PMT components were evident. In-clinic behavior and attendance were influenced by affect, whereas dropout status was predicted by affect, severity, self-efficacy, and age. The screening tool used in this study may assist in identifying patients who are at risk for poor adherence and provide valuable information to enhance provider-patient relationships and foster patient adherence. However, it is recommended that more research be conducted to further understand the impact of variables on patient adherence and that the screening tool be enhanced to increase its predictive ability.


Repair capacity and kinetics of human skin during fractionated radiotherapy: erythema, desquamation, and telangiectasia after 3 and 5 year's follow-up.
Prospective clinical fractionation studies on acute and late reactions in skin have been going on since 1972 at the Radiotherapy Department in Gothenburg. The clinical assay consisted of breast cancer patients irradiated postoperatively to the internal mammary nodes from unilateral or bilateral fields exposed to various dose schedules. 750 fields in 450 patients have been analysed. Schedules with 1, 2 or 5 fractions per week and 2 or 3 fractions per day were evaluated with erythema, desquamation and telangiectasia as endpoints. For some schedules a dose-response relationship was established in a limited dose range, but often there was only one dose group per schedule. These data are suited to analysis by the method of direct analysis of quantal response. This was used in the present analysis, along with the linear quadratic (LQ) model and its generalization, the incomplete repair (IR) model. The repair capacity was similar for erythema and desquamation, with alpha/beta ratios between 7.5 and 11.2 Gy. Unexpectedly, there was more significant time factor during radiotherapy courses up to 6 weeks for erythema and desquamation, but the repair capacity was changed after 4 weeks for both endpoints, and alpha/beta increased to between 18.3 and 34.5 Gy. The repair capacity for late telangiectasia differed significantly from that for erythema and desquamation, with alpha/beta values between 2.8 and 4.3 Gy. There was a significant time factor for telangiectasia with characteristic doubling time of about 16 days, when an exponential function for time was used. Concerning the repair kinetics in skin, there were insufficient data to obtain precise estimates, but there was a suggestion of two components of repair. This was inferred from higher-than-predicted recovery with 15-min intervals, when the data were fitted with the monoexponential model. The monoexponential fit gave t1/2 between 1.1 and 1.3 h for acute effects and 3.5 h for late effects. Recovery after 15-min fractionation intervals, if it resulted from a fast repair component, would be consistent with a half-time of 0.3-0.4 h. The time factor and the relative long half-time for repair for late effects have important implications for multiple-fraction-per-day treatment, and imply that interfraction intervals of 4 h or less, as commonly used, will be insufficient. Instead, intervals of 6 h or longer are recommended. Using accelerated fractionation with a significant reduction in overall treatment time a dose reduction is still necessary to take into account the time factor for late effects.(ABSTRACT TRUNCATED AT 400 WORDS)


Elemental analysis of solid microscopic samples in the flameless atomic absorption cuvette.
Solid microscopic samples are precisely located by means of a newly developed applicator under microscopic control in the center of a flameless graphite tube cuvette. The parts of the applicator that reach into the cuvette are made of quartz which can be cleaned by heating. The values for the K and Na content of samples such applied are highly reproducible. No contamination could be detected. Solid samples yield higher signals than liquid samples. A method of calibration is described which uses lyophilized pieces of egg albumin containing known amounts of K and Na as standards.


Effect of diabetes on the tissular Zn/Cu ratio.
One of the parameters related to the development of coronary disease in diabetic patients is the tissular Zn/Cu ratio. We evaluated the levels of Zn and Cu, and the Zn/Cu ratio in insulin target tissues in diabetic and normoglucemic growing Wistar rats in order to determine the influence of diabetes and the disease evolution period. Diabetes was induced chemically by administration of streptozotocin. In order to determine the influence of the duration of diabetes on the Zn/Cu ratio, three time periods were studied: 7, 21 and 60 days. The animals were subsequently sacrificed and the target tissues (liver, adipose tissue, and skeletal muscle) were removed. Zn and Cu levels were measured by AAS after wet mineralization. STZ-induced diabetes modified the tissular Zn and Cu content. There was a significant decrease (p < 0.01) in liver and adipose tissue, but not in skeletal muscle. The in adipose tissue and skeletal muscle, but not in liver, effects were dependent on the duration of diabetes. The Cu content was higher in the liver of diabetic rats (p < 0.1) and lower in adipose tissue (p < 0.1) and skeletal muscle (NS). Tissular Cu levels also were affected significantly by the duration of diabetes. The Zn/Cu ratio showed a generalized decrease, except in skeletal muscle. This decrease was dependent on the presence of diabetes mellitus and the duration of the disease (p < 0.01).


Alveolar bone remodelling pattern of the rat incisor under different functional conditions as shown by minocycline administration.
Remodelling of the socket surrounding the continuously growing and erupting rat incisor was examined in teeth under normo, hyper and hypofunctional conditions. Cross-sections of the mandible were observed under fluorescence microscopy, where minocycline labelling evidenced bone remodelling. Animals had received minocycline (10 mg/day) during the experimental period. Control animals (from all three groups) received vehicle alone and samples from these animals were not fluorescent. Minocycline did not interfere with the eruption rates in any of the functional conditions studied. Normofunctional (impeded) incisors showed constant osteogenic activity in the alveolar bone facing the periodontal ligament in all regions of the incisor. Under hypofunctional (unimpeded) and hyperfunctional (impeded) conditions, osteogenesis in the region close to the alveolar crest was markedly increased in the mesial wall of the socket. The labial alveolar bone, facing the enamel-related periodontium, was almost entirely formed during the experimental period in all the groups, but in hyper and hypofunctional teeth the newly formed bone was thicker and contained a substantial amount formed before the experimental period. In the more apical regions of the socket no marked differences between the three functional conditions were found. The similar bone remodelling shown in hypo and hyperfunctional teeth might indicate that there are common factors causing this pattern. Consideration of possible factors appear to rule out the eruption rate, which is very different under these two functional conditions.


A new method for the insertion of multiple microprobes into neural and muscular tissue, including fiber electrodes, fine wires, needles and microsensors.
We developed a new method for the insertion of thin-shaft probes into neural and muscular tissue. Axial forces for driving the probes into tissue and radial forces against buckling are both provided by a stretched elastic rubber tube in which the probe is guided outside the tissue. Various geometric arrangements of arrays with independently advanceable probes are possible. Prototypes with 7 linearly aligned fiber electrodes and computer-controlled positioning motors were successfully used in single- and multiple-unit recordings from the visual system of awake monkeys (Eckhorn et al., 1993). The method is suitable in a wide range of applications, including insertion of fine microprobe fibers and wire electrodes into brain and muscle through the skin or dura, provided that the tips of the probes are sharp and hard enough.


Classifying the superfamily of small heat shock proteins by using g-gap dipeptide compositions.
Small heat shock protein (sHSP) is a superfamily of molecular chaperone and is found from archaea to human. Recent researches have demonstrated that sHSPs participate in a series of biological processes and are even closely associated with serious diseases. Since sHSP is a very large superfamily and members from different superfamilies exhibit distinct functions, accurate classification of the subfamily of sHSP will be helpful for unrevealing its functions. In the present work, a support vector machine-based method was proposed to classify the subfamily of sHSPs. In the 10-fold cross validation test, an overall accuracy of 93.25% was obtained for classifying the subfamily of sHSPs. The superiority of the proposed method was also demonstrated by comparing it with the other methods. It is anticipated that the proposed method will become a useful tool for classifying the subfamily of sHSPs.


Hypokalaemia in severe head trauma.
Forty-six consecutive patients with severe isolated head trauma (GCS less than or equal to 7) had significant hypokalaemia averaging 3.1 +/- 0.4 (S.D.) mmol/litre upon admission to the emergency room. This electrolyte imbalance occurred within hours of the trauma and resolved under treatment within the first day. There was no correlation between serum potassium and pH, glucose, urine electrolytes, patient age or sex, admission Glasgow Coma Score or the Glasgow Outcome Score. Sixteen patients with multiple trauma but without head trauma had an average serum potassium of 3.5 +/- 1.1 mmolar. Mechanisms of serum potassium level control and their correlation with brain trauma are addressed in a search for an explanation of this phenomenon. We believe that the most appropriate explanation for this hypokalaemia is the large catecholamine discharge that is known to accompany severe head trauma, with resultant beta 2-adrenergic stimulation of the Na+ -K+ pump. The formation of a prospective on-going study to clarify the basic mechanism of hypokalaemia in severe head trauma is presented.


Intravenous administration of apeling-13 induces a depressor response by releasing an unidentified substance.
Apelin and APJ receptor play an important role in the regulating cardiovascular function; however, conflicting results have been reported regarding the effect of apelin on cardiovascular regulation. In this study, blood pressure and heart rate were measured by femoral arterial catheterization; and cardiac contractility was recorded by left ventricular catheterization through the right carotid artery in rats before and after intravenous administration of [pyr1]-apelin-13. The results show that intravenous administration of apelin-13 caused a dramatic reduction in BP but did not significantly alter heart rate and contractility. To study the mechanism of the apelin-induced depressor response, isometric tension was measured in isolated mesenteric arteries using a myograph approach. Surprisingly, treatment of the arteries with [pyr1]-apelin-13 did not cause relaxation of mesenteric arteries preconstricted with norepinephrine; however, treatment with plasma collected from rats that received intravenous administration of [pyr1]-apelin-13 caused pronounced relaxation of isolated arteries. Incubation with the guanylyl cyclase inhibitor, ODQ, blocked NO-induced relaxation, but did not significantly alter the relaxation response to the plasma from apelin-treated rats. Taken together, these findings demonstrate that intravenous injection of apelin causes a significant depressor response that is mediated by a NO-independent mechanism involving an unidentified substance released into the bloodstream leading to vasodilation.


Altered synchrony of right ventricular contraction in borderline pulmonary hypertension.
Imaging studies have shown that pulmonary hypertension (PH) is associated with inhomogenous right ventricular (RV) regional contraction, or dyssynchrony, and that this is of prognostic relevance. This study aimed at the identification and functional significance of RV dyssynchrony in borderline PH defined by a mean pulmonary artery pressure between (mPAP) 20 and 25 mmHg. RV dyssynchrony was measured by 2-dimensional speckle tracking echocardiography in 17 patients with pulmonary arterial hypertension (PAH), 13 patients with borderline PH and 14 controls. Dyssynchrony was defined as the R-R interval-corrected standard deviation of the times to peak-systolic strain for the basal and medium segments of the RV. All the PH patients underwent a right heart catheterization. RV dyssynchrony amounted to 69 ± 34 ms in PAH, 47 ± 23 ms in borderline PH and 8 ± 6 ms in controls, all different from each other (p < 0.05). RV dyssynchrony in borderline PH was the only parameter of RV systolic dysfunction in 11 of 13 (85%) of the patients. RV dyssynchrony was accompanied by postsystolic shortening and correlated to RV fractional area change, not to mPAP or pulmonary vascular resistance. RV dyssynchrony occurs in borderline PH and may reflect early RV-arterial uncoupling.


Malalignment of the interventricular septum with atrioventricular septal defect: its implications concerning conduction tissue disposition.
Malalignment of the interatrial and interventricular septal structures is rarely seen associated with atrioventricular septal defects. Such malalignment gives rise to an anomalous disposition of the conduction tissue specifically in regard to the siting of the atrioventricular node. Recognition of this abnormality prior to (or at) surgery is important if post-operative rhythm problems are to be avoided.