Effect of iron overload on furin expression in wild-type and β-thalassemic mice.
Furin is a proprotein convertase enzyme. In the liver, it cleaves prohepcidin to form active hepcidin-25, which regulates systemic iron homeostasis. Hepcidin deficiency is a component of several iron overload disorders, including β-thalassemia. Several studies have identified factors that repress hepcidin gene transcription in iron overload. However, the effect of iron overload on furin, a post-translational regulator of hepcidin, has never been evaluated. The present study aimed to investigate the changes in furin and related factors in parenteral iron-overloaded mice, including those with β-thalassemia. Wild-type (WT) and β-thalassemia intermedia (th3/+) C57BL/6 mice were intraperitoneally injected with 9 doses of iron dextran (1 g iron/kg body weight) over 2 weeks. In the iron overload condition, our data demonstrated a significant Furin mRNA reduction in WT and th3/+ mice. In addition, the liver furin protein level in iron-overloaded WT mice was significantly reduced by 70% compared to control WT mice. However, the liver furin protein in iron-overloaded th3/+ mice did not show a significant reduction compared to control th3/+ mice. The hepcidin gene (hepcidin antimicrobial peptide gene, Hamp1) expression was increased in iron-overloaded WT and th3/+ mice. Surprisingly, the liver hepcidin protein level and total serum hepcidin were not increased in both WT and th3/+ mice with iron overload, regardless of the increase in Hamp1 mRNA. In conclusion, we demonstrate furin downregulation in conjunction with Hamp1 mRNA-unrelated pattern of hepcidin protein expression in iron-overloaded mice, particularly the WT mice, suggesting that, not only the amount of hepcidin but also the furin-mediated physiological activity may be decreased in severe iron overload condition.


In silico comparison of the dosimetric impacts of a greater omentum spacer for abdominal and pelvic tumors in carbon-ion, proton and photon radiotherapy.
The purpose of this study was to compare carbon-ion (C-ion), proton and photon radiotherapy (RT) plans with regard to dose reduction of the gastrointestinal (GI) tract by using a greater omentum spacer (GO spacer). We retrospectively retrieved data for ten patients who received the GO spacer as surgical spacer placement for abdominal and pelvic tumors. Simulation plans were created on pre-spacer Computed Tomography (CT) and post-spacer CT for C-ion RT, proton RT and photon RT to compare the dose of the GI tract. The plans were normalized so that at least 95% of the planning target volume (PTV) received 70 Gy (relative biological effectiveness equivalent) delivered in 35 fractions. All plans were created with the lowest possible dose to the GI tract under conditions that meet the dose constraints for the PTV and spinal cord (maximum dose < 45 Gy). The part of the GI tract to be evaluated was defined as that most adjacent to the PTV. C-ion RT plans and proton RT plans were calculated by a spot scanning technique, and photon RT plans were calculated employing by fixed-field intensity-modulated radiation therapy. D2 cc and V10-70 of the GI tract were significantly lower on post-spacer plans than on pre-spacer plans for all three RT modalities. Regarding post-spacer plans, D2 cc of the GI tract was significantly lower on C-ion RT plans and proton RT plans than on photon RT plans (C-ion vs photon p = 0.001, proton vs photon p = 0.002). However, there was no significant difference between C-ion RT plans and proton RT plans for D2 cc of the GI tract (C-ion vs proton p = 0.992). In the photon RT plan for one patient, D2 cc of the GI tract did not meet < 50 Gy. The GO spacer shows a significant dose reduction effect on the GI tract.


Effect of preoperative radiochemotherapy on lymph node retrieval after resection of rectal cancer.
Preoperative radiochemotherapy for advanced rectal cancer results in fewer lymph nodes detected in the tumor-bearing specimen. Nonrandomized control trial with analysis of a prospective perioperative database. Department of Surgery of a large-volume university hospital. All patients who underwent conventional open surgery to cure rectal cancer between January 1, 1996, and March 31, 2001. During the study period 184 patients (81%, control group) underwent surgery without receiving preoperative radiochemotherapy. Forty-two patients (19%, study group) who had advanced rectal cancer (modified Dukes stages B [tumors that have penetrated the muscle layer of the bowel wall or have gone through the bowel] or C [tumors that have spread to the lymph nodes in the same region]) received preoperative radiochemotherapy (2 cycles of fluorouracil, 4500 rad) during this period. Most patients underwent anterior rectal resection in both groups (77.7% of those who did not receive preoperative radiochemotherapy and 71.8% of those who did), the remaining patients were treated with abdominoperineal resection. A mean (SEM) of 19 (1) lymph nodes per specimen were detected in the control patients, while significantly fewer lymph nodes were detected in study patients (13 [1]; P<.05). The rate of inadequate lymph node staging (pNx) increased from 7% in the control group to 12% in the study group (P =.06). Pathological lymph node staging disclosed that significantly more study patients who received preoperative radiochemotherapy had modified Dukes stage A (tumors that are found only in the inner wall or rectum) cancer when compared with the control group (17% vs 0%, respectively; P<.05). Preoperative radiochemotherapy for advanced rectal cancer results in a significant decrease of lymph nodes detected within the tumor-bearing specimen. Preoperative radiochemotherapy induces significant downstaging with fewer positive lymph nodes and more patients presenting with Dukes stage A rectal cancer. Great care must be taken to remove an adequate number of lymph nodes and more sophisticated pathological techniques of lymph node detection are required since the tumors of ever-increasing numbers of patients are inadequately classified.


[Fighting melanoma together].
Patients and their families often become involved in non-profit organisations which undertake actions to combat a disease. In the case of melanoma, in addition to promoting research, associations work to encourage prevention and early detection.


[Empty sella syndrome: a case report].
Empty sella syndrome is a condition in which the sella turcica is partially or totally filled with cerebrospinal fluid causing a displacement of the pituitary gland. We here report the case of a 49-year old obese patient with progressive headaches, physical asthenia and hypothyroidism. Brain scanner showed empty sella syndrome.


Importance of metabolites in antiarrhythmic therapy.
The presence of metabolites with pharmacologic activity can produce unanticipated drug efficacy or toxicity. This is particularly true during treatment with drugs that have narrow therapeutic-toxic ratios, such as the antiarrhythmic agents. The presence of active metabolites can often be inferred from variability in the relation between pharmacologic effect and steady-state plasma concentrations of the parent drug. Moreover, metabolites may ordinarily be unimportant but can accumulate to therapeutic (or toxic) levels in disease states such as congestive heart failure, renal failure and hepatic failure. Further characterization of the contribution of such metabolites during treatment requires direct evaluation of their pharmacology in vitro, in animal models and, if indicated, in man. Procainamide and its active metabolite N-acetylprocainamide provide the best and most complete example of this sequence of observations. Other drugs, including quinidine, disopyramide, verapamil and the investigational agents encainide and lorcainide, have active metabolites for which pharmacologic activity is less well-defined. Further studies in this area will help reduce the frequency of antiarrhythmic drug adverse effects, make successful therapy more frequent, and perhaps allow insights into structure-activity relations.


X-linked genetic homologies between mouse and man.
X-linked genes are conserved among all mammalian species, but the organization of genes on the X chromosome varies from one species to another. This review summarizes the evidence for established gene homologies between mice and human beings. It also describes genes that are possible homologies because of their locations in the human and murine X chromosomes and similarities in the phenotypes they produce. Based on current knowledge of homologous gene location, the human and murine X chromosomes appear to contain four highly conserved segments and differ in organization by only three to four simple chromosomal rearrangements.


Long-term distress after radical prostatectomy versus watchful waiting in prostate cancer: a longitudinal study from the Scandinavian Prostate Cancer Group-4 randomized clinical trial.
Studies enumerating the dynamics of physical and emotional symptoms following prostate cancer (PCa) treatment are needed to guide therapeutic strategy. Yet, overcoming patient selection forces is a formidable challenge for observational studies comparing treatment groups. To compare patterns of symptom burden and distress in men with localized PCa randomized to radical prostatectomy (RP) or watchful waiting (WW) and followed up longitudinally. The three largest, Swedish, randomization centers for the Scandinavian Prostate Cancer Group-4 trial conducted a longitudinal study to assess symptoms and distress from several psychological and physical domains by mailed questionnaire every 6 mo for 2 yr and then yearly through 8 yr of follow-up. RP compared with WW. A questionnaire was mailed at baseline and then repeatedly during follow-up with questions concerning physical and mental symptoms. Each analysis of quality of life was based on a dichotomization of the outcome (yes vs no) studied in a binomial response, generalized linear mixed model. Of 347 randomized men, 272 completed at least five questionnaires during an 8-yr follow-up period. Almost all men reported that PCa negatively influenced daily activities and relationships. Health-related distress, worry, feeling low, and insomnia were consistently reported by approximately 30-40% in both groups. Men in the RP group consistently reported more leakage, impaired erection and libido, and fewer obstructive voiding symptoms. For men in the WW group, distress related to erectile symptoms increased gradually over time. Symptom burden and distress at baseline was predictive of long-term outlook. Cancer negatively influenced daily activities among almost all men in both treatment groups; health-related distress was common. Trade-offs exist between physiologic symptoms, highlighting the importance of tailored treatment decision-making. Men who are likely to experience profound long-term distress can be identified early in disease management.


MRI of the ulnar nerve pre- and post-transposition: imaging features and rater agreement.
Determine the rater agreement of MRI features of the ulnar nerve pre- and post-transposition and association with recurrent symptoms. This IRB-approved retrospective cohort analysis examined 23 subjects who underwent elbow MRI pre- and post-ulnar nerve transposition from 1999 to 2018, 10 of whom developed recurrent symptoms. Pre- and post-transposition MRIs were evaluated by two blinded radiologists for ulnar nerve cross-sectional area, signal intensity, fascicular architecture, caliber change, and perineural scar. Inter-rater agreement was estimated using intraclass correlation coefficients (ICCs) for continuous variables and Gwet's agreement coefficient (AC) for categorical variables. Binary logistic regression modeling probed associations between imaging markers and symptom recurrence. The ulnar nerve, post-operatively, demonstrated statistically significant increases in size (p < 0.001), signal intensity (p = 0.021), and abrupt caliber change (p = 0.024). None of the imaging features, except for higher signal intensity of the nerve pre-transposition as demonstrated by one rater, were predictive of symptom recurrence. Inter-rater agreement for cross-sectional area measurements of the ulnar nerve at the cubital tunnel was excellent (ICCs of 0.91 and 0.83). Substantial-to-excellent inter-rater agreement was observed pre-operatively for nerve signal intensity, caliber change, and fascicular architecture. Post-operatively, agreement on nerve signal intensity and perineural scar was excellent (ACs of 0.90 and 0.88), but only slight for caliber change (0.15). Inter-rater agreement for qualitative and quantitative assessment of the ulnar nerve was generally robust. Post-transposition, the ulnar nerve was generally larger and more T2-hyperintense, but MRI features were not predictive of recurrent ulnar neuropathy, except for perhaps the signal intensity of the nerve pre-transposition.


Rickettsia japonica infections in Huanggang, China, in 2021.
Two patients from Huanggang, China, were diagnosed with spotted fever group (SFG) rickettsiosis-caused by spotted fever group rickettsiae (SFGR)-in 2021. This study aimed to investigate the clinical symptoms, laboratory examinations, epidemiological factors, and therapeutic responses in patients with SFG rickettsiosis-an emerging disease in this region. The patients showed a variety of clinical signs and symptoms, such as acute febrile illness with severe headache, myalgia, asthenia, anorexia, eschar, lymphadenopathy, and rash on the trunk and extremities. They exhibited increased neutrophil ratio, mild thrombocytopenia, liver dysfunction, and increased C-reactive protein and procalcitonin levels. Following treatment with doxycycline, the patients recovered completely. This is the first report of Rickettsia japonica infection in Huanggang City, Hubei Province, China. SFGR infection is a tick-borne disease, which can be effectively treated with doxycycline; however, it has a mortality rate of approximately 10% with delays in treatment. The Huanggang area is also a high-risk area for tick-borne severe fever with thrombocytopenia syndrome (SFTS). Therefore, SFTS and SFG rickettsiosis should be carefully diagnosed in this area and clinicians should be alert with respect to the possibility of infections with both SFTS and SFG rickettsiosis.


Biochemical markers of bone turnover. I: Theoretical considerations and clinical use in osteoporosis.
The recent development of noninvasive techniques to measure bone mass and bone turnover represents a major advance in the diagnosis and management of osteoporosis. The rate of formation or degradation of the bone matrix can be assessed either by measuring a prominent enzymatic activity of the bone-forming or resorbing cells, such as alkaline and acid phosphatase activity, or by measuring bone matrix components released into the circulation during formation or resorption. Recent studies have shown that the appropriate combination of the most efficient markers of bone resorption and formation will provide a powerful tool in the clinical investigation of osteoporosis.


Pharmacological characterization and neutralization of the venoms used in the production of Bothropic antivenom in Brazil.
Some pharmacological effects of the venoms of five different Brazilian Bothrops species and a pool of these venoms (AgB) were quantified. The ability of polyspecific Bothropic antivenom produced at Fundação Ezequiel Dias (FUNED, Brazil) to neutralize the principal toxic and enzymatic activities was studied using in vivo and in vitro assays. The lethality, hemorrhagic, necrotizing, proteolytic, phospholipase, coagulant and fibrinolytic activities were measured for each of these venoms. Comparison of protein electrophoretic patterns showed significant differences such as the presence of common and also unique components. Furthermore, experimental studies revealed differences in their biological properties among individual samples. It was found that the Bothrops antivenom was highly effective in the neutralization of the toxic activities of all venoms tested. In addition, indirect ELISA was used to compare the antigenic cross-reactivity for each of the five Bothrops venoms as well as the venoms of B. atrox, B. leucurus and B. erythromelas which were not included in the antigenic pool (AgB). Therefore, the characterization of several toxic activities of snake venoms is necessary, if toxicity is to be properly evaluated. Results indicate that the Brazilian antivenom prepared at FUNED against Bothrops snakes is effective in neutralizing the main toxic effects of Bothrops venoms.


Surgical Therapies for Vitiligo.
Surgical management is a safe and effective treatment modality for select patients with vitiligo. Many techniques of vitiligo surgery exist, each with unique advantages and disadvantages. Preoperative screening for appropriate candidates, selection of surgical technique, and postoperative management are all key elements in enabling patients to achieve maximal repigmentation.


Integration of proteome and transcriptome refines key molecular processes underlying oil production in Nannochloropsis oceanica.
Under nitrogen deficiency situation, Nannochloropsis spp. accumulate large amounts of lipids in the form of triacylglycerides (TAG). Mechanisms of this process from the perspective of transcriptome and metabolome have been obtained previously, yet proteome analysis is still sparse which hinders the analysis of dynamic adaption to nitrogen deficiency. Here, proteomes for 3 h, 6 h, 12 h, 24 h, 48 h and 10th day of nitrogen deplete (N-) and replete (N+) conditions were obtained and integrated with previous transcriptome data for N. oceanica. Physiological adaptations to N- not apparent from transcriptome data were unveiled: (a) abundance of proteins related to photosynthesis only slightly decreased in the first 48 h, indicating that photosynthesis is still working efficiently, and protein amounts adjust gradually with reduction in chloroplast size. (b) Most proteins related to the TCA cycle were strongly upregulated after 48 h under N-, suggesting that respiration is enhanced after 48 h and that TCA cycle efflux supports the carbon required for lipid synthesis. (c) Proteins related to lipid accumulation via the Kennedy pathway increased their abundance at 48 h, synchronous with the previously reported diversification of fatty acids after 48 h. This study adds a proteome perspective on the major pathways for TAG accumulation in Nannochloropsis spp. Temporal changes of proteome exhibited distinct adaptation phases that are usually delayed relative to transcriptomic responses. Notably, proteome data revealed that photosynthesis and carbon fixation are still ongoing even after 48 h of N-. Moreover, sometimes completely opposite trends in proteome and transcriptome demonstrate the relevance of underexplored post-transcriptional regulation for N- adaptation.


Diagnostic application of total antioxidant capacity in seminal plasma to assess oxidative stress in male factor infertility.
This study was undertaken in order to establish a new reference value for the total antioxidant capacity (TAC) in seminal plasma as a predictor of fertility. This study also aims to propose a detailed protocol for the TAC assay including calculation of assay results and assessment of sensitivity and specificity over possible cutoff values in infertile men and controls with proven and unproven fertility. Seminal plasma from 279 infertile patients and 46 normal healthy men referred to a male infertility testing laboratory were tested to measure TAC by a colorimetric assay kit. Receiver operating characteristics (ROC) curves were generated to establish cutoff values, sensitivity, and specificity, and the distribution of cutoff values in controls and infertile patients was calculated. Infertile patients showed significantly lower levels (mean ± SEM) of total antioxidants (micromolar Trolox equivalents) in their seminal plasma (1863.84 ± 27.16 μM) compared to those from fertile men (2013 ± 56.04 μM, P = 0.019). A preferred cutoff TAC value of 1947 μM could facilitate better diagnosis of oxidative stress (OS) in men with male factor infertility. At this threshold, the specificity of TAC assay was 63.0 % and the sensitivity 59.5 % with a positive predictive value of 90.7 % and a negative predictive value of 20.4 %. Our results establish a new diagnostic cutoff TAC value of 1947 μM in seminal plasma to distinguish prevalence of OS in infertile patients compared to healthy men. This study provides a robust reference value of seminal plasma TAC that may provide an important diagnostic tool to the physicians for managing OS and male factor infertility in such patients.


Interplay Between Inflammatory-immune and Interleukin-17 Signalings Plays a Cardinal Role on Liver Ischemia-reperfusion Injury-Synergic Effect of IL-17Ab, Tacrolimus and ADMSCs on Rescuing the Liver Damage.
This study tested the hypothesis that inflammatory and interleukin (IL)-17 signalings were essential for acute liver ischemia (1 h)-reperfusion (72 h) injury (IRI) that was effectively ameliorated by adipose-derived mesenchymal stem cells (ADMSCs) and tacrolimus. Adult-male SD rats (n = 50) were equally categorized into groups 1 (sham-operated-control), 2 (IRI), 3 [IRI + IL-17-monoclonic antibody (Ab)], 4 (IRI + tacrolimus), 5 (IRI + ADMSCs) and 6 (IRI + tacrolimus-ADMSCs) and liver was harvested at 72 h. The main findings included: (1) circulatory levels: inflammatory cells, immune cells, and proinflammatory cytokines as well as liver-damage enzyme at the time point of 72 h were highest in group 2, lowest in group 1 and significantly lower in group 6 than in groups 3 to 5 (all p < 0.0001), but they did not differ among these three latter groups; (2) histopathology: the liver injury score, fibrosis, inflammatory and immune cell infiltration in liver immunity displayed an identical pattern of inflammatory cells among the groups (all p < 0.0001); and (3) protein levels: upstream and downstream inflammatory signalings, oxidative-stress, apoptotic and mitochondrial-damaged biomarkers exhibited an identical pattern of inflammatory cells among the groups (all p < 0.0001). Our results obtained from circulatory, pathology and molecular-cellular levels delineated that acute IRI was an intricate syndrome that elicited complex upstream and downstream inflammatory and immune signalings to damage liver parenchyma that greatly suppressed by combined tacrolimus and ADMSCs therapy.


Determination of the complete cDNA sequence of rat type II collagen and evaluation of distinct expression patterns of types IIA and IIB procollagen mRNAs during fracture repair in rats.
Elucidating the molecular mechanisms that underlie fracture healing is crucial to understanding and devising strategies for the management of fractures, especially those associated with a pathological condition such as diabetes or old age. Cartilage formation, and therefore the expression of type II collagen by chondrocytes, is a critical step in frac-ture healing. Two forms of type II collagen, IIA and IIB, are known to be produced by alternative splicing of the Alpha(1) (II) procollagen gene. We have followed the patterns of expression of these two forms of type II collagen to determine the nature of chondrocyte recruitment during fracture healing. First, we sequenced the rat collagen type II cDNA to design the primers. Second, using a competitive quantitative reverse transcription-mediated polymerase chain reaction, we provide evidence that (1) there is a basal level of type IIA collagen expression during the early stages of fracture healing; (2) transient but sharp up-regulation of IIA expression occurs concomitant with chondrogenesis and endochondral ossification; and (3) type IIB collagen is the predominant mRNA variant expressed at virtually all times during fracture repair.


The role of pre-operative assessment and ringfencing of services in the control of methicillin resistant Staphlococcus aureus infection in orthopaedic patients.
MRSA is a major economic and health issue internationally and as such is of particular importance in the appropriate management of orthopaedic patients. Bone, joint and implant infection can lead to unfavourable outcomes with a long protracted in hospital stay inevitable. The cost for the patient, the hospital and society are substantial. This study was a review of a prospectively maintained database from our unit over three time points from 2005 to 2007. At each time point a new infection control measure was implemented in an effort to reduce MRSA infections. Total rates of MRSA infection and colonisation in all orthopaedic patients were recorded, before and after separation of trauma and elective services, and after the introduction of a screening pre assessment clinic. 12259 orthopaedic patients were reviewed over the three years. The mean age of MRSA infected patients was 71. A higher proportion of female patients were infected than male patients. The mean length of stay for infected patients was 23.4 days. The rate of infection dropped from 0.49% in 2005 to 0.24%in 2007. After the introduction of these measures there was a substantial reduction in organ space and deep tissue infections. The separation of emergency and elective orthopaedic services coupled with effective pre-operative screening has resulted in a significant reduction in MRSA infection despite an ever increasing prevalance.


Differential recognition of the serologically defined HLA-A2 antigen by allogeneic cytotoxic T cells. II. Definition of three HLA-A2 subtypes by CTLs.
A comprehensive analysis of human alloimmune cytotoxic T lymphocytes (CTLs) specific for the HLA-A2 antigen identified 11% of HLA-A2 positive cells as outliers. In total, 11 unrelated serologically indistinguishable, but distinguishable by cell-mediated lympholysis (CML) HLA-A2 positive outlier cells were identified. The outlier cells could be subdivided in two subgroups according to reactivity patterns obtained with CTLs directed against the HLA-A2 antigen of outlier cells and their inhibitory capacity in specific competitive inhibition experiments. Thus, the serologically defined HLA-A2 specificity can be divided into at least three subtypes using CTLs specific for the HLA-A2 antigen. Moreover, CTLs specific for an HLA-A2 subtype could be induced when responder cells expressed a different HLA-A2 subtype antigen. On the basis of several family studies, we conclude that the subtype HLA-A2 antigens are inherited in a codominant way.


3D genomics imposes evolution of the domain model of eukaryotic genome organization.
The hypothesis that the genome is composed of a patchwork of structural and functional domains (units) that may be either active or repressed was proposed almost 30 years ago. Here, we examine the evolution of the domain model of eukaryotic genome organization in view of the expansion of genome-scale techniques in the twenty-first century that have provided us with a wealth of information on genome organization, folding, and functioning.


Simultaneous stabilization of Pb, Cd, and As in soil by rhamnolipid coated sulfidated nano zero-valent iron: Effects and mechanisms.
Sulfidation effectively improves the electron transfer efficiency of nanoscale zero-valent iron (nZVI), but decreases the specific surface area of nZVI. In this study, sulfidated nZVI (S-nZVI) coated with rhamnolipid (RL-S-nZVI) was synthesized and used to stabilize Pb, Cd, and As in combined polluted soil. The stabilization efficiency of 0.3% (wt) RL-S-nZVI to water soluble Pb, Cd, and As in soil reached 88.76%, 72%, and 63%, respectively. Rhamnolipid coating inhibited the reduction of specific surface area and successfully encapsulated nZVI, thus reducing the oxidation of Fe0. The types of iron oxides in RL-S-nZVI were reduced compared to S-nZVI, but the content and strength of Fe0 iron were obviously enhanced. Furthermore, rhamnolipid functional groups (-COOH and -COO-) were also involved in the stabilization process. In addition, the stabilization efficiency of RL-S-nZVI to the bioavailable Pb, Cd, and As in soil increased by 41%, 41%, and 50%, respectively, compared with nZVI. The presence of organic acids, especially citric acid, improved the stabilization efficiency of RL-S-nZVI to the three metals. The result of BCR sequential extraction indicated that RL-S-nZVI increased the residual state of Pb, Cd, and As and reduced the acid-soluble and reducible state after 28 days of soil incubation. XRD and XPS analyses showed that the stabilization mechanisms of RL-S-nZVI on heavy metals involved in ion exchange, surface complexation, adsorption, co-precipitation, chemisorption, and redox.


Biological and novel therapies for inflammatory bowel disease in children.
During the past decade, a tremendous wealth of information regarding the pathogenesis, genetics, and therapy of IBD has been discovered. Judging by the number of new publications published every month in scientific journals and the great enthusiasm at scientific meetings, this outstanding pace surely will continue. In the near future, clinicians may be able to classify IBD into several subtypes depending on patients' cytokine and gene profiles. For example, two groups of researchers recently have identified mutation in the NOD2 gene, which is associated with susceptibility to CD. This identification may allow the clinician to better predict outcome and response to medical therapy. At the same time, several promising new therapies are being investigated. Technologic advances will continue to result in the development of potent and specific agents that will control and possibly correct the abnormal inflammatory processes responsible for pediatric IBD.


Rationally designed carbohydrate-occluded epitopes elicit HIV-1 Env-specific antibodies.
An array of carbohydrates masks the HIV-1 surface protein Env, contributing to the evasion of humoral immunity. In most HIV-1 isolates 'glycan holes' occur due to natural sequence variation, potentially revealing the underlying protein surface to the immune system. Here we computationally design epitopes that mimic such surface features (carbohydrate-occluded neutralization epitopes or CONE) of Env through 'epitope transplantation', in which the target region is presented on a carrier protein scaffold with preserved structural properties. Scaffolds displaying the four CONEs are examined for structure and immunogenicity. Crystal structures of two designed proteins reflect the computational models and accurately mimic the native conformations of CONEs. The sera from rabbits immunized with several CONE immunogens display Env binding activity. Our method determines essential structural elements for targets of protective antibodies. The ability to design immunogens with high mimicry to viral proteins also makes possible the exploration of new templates for vaccine development.


Chemokines and common variable immunodeficiency; possible contribution of the fractalkine system (CX3CL1/CX3CR1) to chronic inflammation.
Common variable immunodeficiency (CVID) is a heterogeneous syndrome characterized by defective immunoglobulin production and high frequency of bacterial infections, autoimmunity and manifestations of chronic inflammation. The chemokine Fractalkine (CX3CL1) and its receptor CX3CR1 is suggested to play an important role in the pathogenesis of several inflammatory disorders. We hypothesized that enhanced CX3CL1/CX3CR1 interaction could be involved in the chronic inflammation characterising subgroups of CVID. CVID patients were characterized by raised plasma levels of CX3CLl and enhanced expression of its corresponding receptor CX3CR1 on CD4(+) and CD8(+) T cells, including both CD45RA(+) and CD45RA(-) subsets. CX3CR1 expression was particularly enhanced in patients characterized by chronic inflammation in vivo. The high expression of the receptor in CVID patients was accompanied by enhanced chemotactic, adhesive, and other inflammatory cell responses to stimulation with CX3CL1. Our findings suggest that increased CX3CL1/CX3CR1 interaction could contribute to the inflammatory phenotype seen in subgroups of CVID patients.


Interplay between viruses and host sumoylation pathways.
Post-translational modification by members of the small ubiquitin-like modifier (SUMO) family of proteins is important for the regulation of many cellular proteins and pathways. As obligate parasites, viruses must engage with the host cell throughout their replication cycles, and it is therefore unsurprising that there are many examples of interplay between viral proteins and the host sumoylation system. This article reviews recent advances in this field, summarizing information on sumoylated viral proteins, the varied ways in which viruses engage with SUMO-related pathways, and the consequences of these interactions for viral replication and engagement with innate and intrinsic immunity.


Weak founder effects but significant spatial genetic imprint of recent contraction and expansion of European beech populations.
Understanding the ecological and evolutionary processes occurring during species range shifts is important in the current context of global change. Here, we investigate the interplay between recent expansion, gene flow and genetic drift, and their consequences for genetic diversity and structure at landscape and local scales in European beech (Fagus sylvatica L.) On Mont Ventoux, South-Eastern France, we located beech forest refugia at the time of the most recent population minimum, ~150 years ago, and sampled 71 populations (2042 trees) in both refugia and expanding populations over an area of 15,000 ha. We inferred patterns of gene flow and genetic structure using 12 microsatellite markers. We identified six plots as originating from planting, rather than natural establishment, mostly from local genetic material. Comparing genetic diversity and structure in refugia versus recent populations did not support the existence of founder effects: heterozygosity (He = 0.667) and allelic richness (Ar = 4.298) were similar, and FST was low (0.031 overall). Still, significant spatial evidence of colonization was detected, with He increasing along the expansion front, while genetic differentiation from the entire pool (βWT) decreased. Isolation by distance was found in refugia but not in recently expanding populations. Our study indicates that beech capacities for colonization and gene flow were sufficient to preserve genetic diversity despite recent forest contraction and expansion. Because beech has long distance pollen and seed dispersal, these results illustrate a 'best case scenario' for the maintenance of high genetic diversity and adaptive potential under climate-change-related range change.


Decreased expression of DMPK: correlation with CTG repeat expansion and fibre type composition in myotonic dystrophy type 1.
Myotonic dystrophy type 1 (DM1) is an autosomal dominant disease caused by a trinucleotide repeat-expansion, cytosine-thymine-guanine (CTG)n, in the 3' untranslated region of a gene encoding the myotonic dystrophy protein kinase (DMPK). To correlate CTG expansion and protein expression, we studied muscle specimens from 16 adult DM1 patients using three anti-DMPK antibodies for immunoblotting. We estimated the amount of the full-length DMPK (85 kDa) in muscle biopsies from normal controls and from DM1 patients carrying different (CTG)n expansions. We found that DMPK concentration was decreased to about 50% in DM patients' muscles; the protein decrease did not seem correlated with the CTG repeat length. However, the fibre type composition in skeletal muscle seemed somehow to affect DMPK decrease, as the lowest level of the enzyme was found in patients with the lowest content of type 1 fibre.


Correction to: Three-dimensional morphometric analysisreveals time-dependent structural changesin microglia and astrocytes in the centralamygdala and hypothalamicparaventricular nucleus of heart failure rats.
An amendment to this paper has been published and can be accessed via the original article.


miRNAs: novel regulators of autoimmunity-mediated pancreatic β-cell destruction in type 1 diabetes.
MicroRNAs (miRNAs) are a series of conserved, short, non-coding RNAs that modulate gene expression in a posttranscriptional manner. miRNAs are involved in almost every physiological and pathological process. Type 1 diabetes (T1D) is an autoimmune disease that is the result of selective destruction of pancreatic β-cells driven by the immune system. miRNAs are also important participants in T1D pathogenesis. Herein, we review the most recent data on the potential involvement of miRNAs in T1D. Specifically, we focus on two aspects: the roles of miRNAs in maintaining immune homeostasis and regulating β-cell survival and/or functions in T1D. We also discuss circulating miRNAs as potent biomarkers for the diagnosis and prediction of T1D and investigate potential therapeutic approaches for this disease.


Bacterial Oxidases of the Cytochrome bd Family: Redox Enzymes of Unique Structure, Function, and Utility As Drug Targets.
Significance: Cytochrome bd is a ubiquinol:oxygen oxidoreductase of many prokaryotic respiratory chains with a unique structure and functional characteristics. Its primary role is to couple the reduction of molecular oxygen, even at submicromolar concentrations, to water with the generation of a proton motive force used for adenosine triphosphate production. Cytochrome bd is found in many bacterial pathogens and, surprisingly, in bacteria formally denoted as anaerobes. It endows bacteria with resistance to various stressors and is a potential drug target. Recent Advances: We summarize recent advances in the biochemistry, structure, and physiological functions of cytochrome bd in the light of exciting new three-dimensional structures of the oxidase. The newly discovered roles of cytochrome bd in contributing to bacterial protection against hydrogen peroxide, nitric oxide, peroxynitrite, and hydrogen sulfide are assessed. Critical Issues: Fundamental questions remain regarding the precise delineation of electron flow within this multihaem oxidase and how the extraordinarily high affinity for oxygen is accomplished, while endowing bacteria with resistance to other small ligands. Future Directions: It is clear that cytochrome bd is unique in its ability to confer resistance to toxic small molecules, a property that is significant for understanding the propensity of pathogens to possess this oxidase. Since cytochrome bd is a uniquely bacterial enzyme, future research should focus on harnessing fundamental knowledge of its structure and function to the development of novel and effective antibacterial agents.


Acute pancreatitis owing to very high triglyceride levels treated with insulin and heparin infusion.
Hypertriglyceridaemia is the third most common cause of acute pancreatitis in the USA. The treatment approach for hypertriglyceridaemia to date has largely been conservative including weight loss, exercise and avoidance of medications that raise triglyceride levels. This approach, however, is not practical in cases of acute pancreatitis due to severely elevated triglycerides. A small number of case reports have been published supporting the treatment of acute pancreatitis due to severely elevated triglyceride levels with insulin and heparin. We report a case of acute pancreatitis in a young woman due to a triglyceride level of 15,215 mg/dl who was successfully treated with insulin and heparin.


New insights into the degradation and detoxification of methylene blue using heterogeneous-Fenton catalyzed by sustainable siderite.
The huge application of synthetic dyes caused a severe impact in the environment. In the present study, a physico-chemical strategy of heterogeneous-Fenton catalyzed by the natural ferrous ore has been established for toxic chemical degradation, of which the complex and high-expense repetitive pH adjustment procedures were escaping. And this natural heterogeneous catalyst also could be recycled and sustainable for toxic substances treatment involved in synergetic adsorption and oxidation. The siderite, served as an adsorbent and catalyst for the degradation of methylene blue (MB). Siderite exhibited a better adsorption capacity with a saturated adsorption capacity of ∼11.08 mg/g. Batch adsorption experiments have verified that adsorption rate and adsorption equilibrium followed pseudo-second-order rate model and Langmuir isotherm equation, respectively. The combination with H2O2, showed significant enhancement of MB degradation without any pH adjustment. The effect of siderite dosage, H2O2 dosage, MB concentration, initial pH, and reaction temperature on MB degradation was investigated, which also has indicated the excellent catalytic performance of siderite. About 99.71% of MB was degraded in 480 min with initial pH of 7.0, reaction temperature of 25 °C, siderite, and H2O2 dosage of 2.5 g/L and 122.38 mM, respectively. It was found that siderite could be reused and remained high degradation efficiency on MB after 5 times reutilization, which also could demonstrate the sustainable and effective process to degrade organic pollution. The generation of reactive species including ·OH and O2·- have been confirmed based on scavenger test and electron spin resonance (ESR) analysis, which was dominated by heterogeneous reaction. The possible degradation mechanisms of MB have been predicted based on spectrum scanning and GC-MS analysis. Moreover, acute toxicity assessment with marine photobacterium Vibrio fisheri was conducted to investigate the toxicity change in the adsorption/oxidation coupled process. This sustainable heterogeneous-Fenton technology has been verified as a promising and applicable process for toxic organic chemicals removal due to effective mineralization and detoxification assisted with the natural ore mineral through the simple operation and mild condtions.


Scanning electron microscopy evaluation of enamel surfaces using different air-polishing powders in the orthodontic setting: an in vitro study.
The aim of this in vitro study was to quantify and compare changes of the enamel surface caused by periodical use of different air-polishing powders during multibracket therapy. Bovine high-gloss polished enamel specimens were air-polished using an AIR-FLOW® Master Piezon with maximum powder and water settings. Each specimen was blasted with sodium bicarbonate (AIR-FLOW® Powder Classic, Electro Medical Systems, Munich, Germany) and erythritol (AIR-FLOW® Powder Plus, Electro Medical Systems). Blasting duration was adapted to the powders' cleaning efficacy and corresponded to 25 air-polishing treatments in a patient with braces. A spindle apparatus ensured uniform guidance at a distance of 4 mm and a 90° angle. Qualitative and quantitative assessments were performed with the use of low vacuum scanning electron microscopy. Following external filtering and image processing, arithmetical square height (Sa) and root mean square height (Sq) were determined. Both prophy powders caused a significant increase in enamel roughness. Surfaces blasted with sodium bicarbonate (Sa = 64.35 ± 36.65 nm; Sq = 80.14 ± 44.80 nm) showed significantly (p < 0.001) higher roughness than samples treated with erythritol (Sa = 24.40 ± 7.42 nm; Sq = 30.86 ± 9.30 nm). The observed defects in enamel structure caused by sodium bicarbonate extended across prism boundaries. Prism structure remained intact after air-polishing with erythritol. Both applied air-polishing powders led to surface alterations. Despite shorter treatment times, sodium bicarbonate was significantly more abrasive than erythritol. Clinicians must compromise between saving time and abrasively removing healthy enamel.


Thymidylate synthase is localized to the nuclear periphery in the yeast Saccharomyces cerevisiae.
To date, the organization of DNA precursor synthesis within eukaryotic cells remains unresolved. Previous studies have suggested the existence of a multienzyme complex that is responsible for DNA precursor synthesis and is associated with sites of replication within the nucleus. Contrasting this, other studies have proposed that DNA precursor synthesis occurs outside the nucleus. To further these studies, we have addressed the location where thymidylate synthase resides in yeast. Subcellular fractionation experiments indicate thymidylate synthase is associated with purified nuclei. Consistent with this, immunofluorescence analysis suggests that thymidylate synthase is situated at the nuclear periphery.


Widely targeted metabolomic analysis reveals that volatile metabolites in cigar tobacco leaves dynamically change during fermentation.
Changes in volatile metabolites during cigar tobacco leaves fermentation as well as the metabolic pathways of metabolites with significant differences were investigated to determine the influence of cigar tobacco leaves fermentation on its flavor. The volatile substances in cigar tobacco leaves at different stages were detected by headspace-solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS), and the main differences in volatile substances in cigar tobacco leaves at different fermentation stages of Yunxue1 in Yuxi production area were analyzed by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). The results show that in the process of cigar tobacco leaves fermentation (YXF0, YXF1, YXF2, YXF3, YXF4, YXF5), a total of 613 volatile metabolites were detected, and a significant difference was found in 263 kinds of metabolites. Among them, the main upregulated differential metabolites were 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, [S-(E,Z,E,E)]-, Benzoic acid, Benzaldehyde, etc. While the main downregulated differential metabolites included beta.-Myrcene, trans-Farnesol, etc. The metabolites with significant differences are mainly concentrated in the biosynthesis of monoterpenes, diterpenes, sesquiterpenes and triterpenes, the degradation metabolism of amino acids, such as valine, leucine and isoleucine, and the biosynthesis of phenylpropyl. There were 8 different metabolites in 5 groups, including 4- (1-methylethyl) -1-cyclohexene-1-formaldehyde、2, 4-dihydroxyacetophenone、2-methylbutyl 3-methylbutyrate and methylpyrazine, all of which showed upregulation trend during fermentation. In the fermentation process, volatile metabolites participate in various synthesis and degradation pathways. The biosynthesis pathway of terpenes and amino acid synthesis and degradation pathway are connected to produce various terpenes, aldehydes and other substances, such as 1,3,6,10-Cyclotetradecatetraene, 3,7,11-trimethyl-14-(1-methylethyl)-, [S-(E,Z,E,E)]-、benzaldehyde and 4-hydroxybenzaldehyde, which are conducive to the overall flavor and quality of cigar tobacco leaves.


Upward Translation of Cerebellar Tonsils following Surgical Expansion of Supratentorial Cranial Vault: A Unified Biomechanical Explanation of Chiari Type I.
Cerebellar tonsils moved significantly upward in 3 patients with Chiari type I who underwent supratentorial cranial vault expansion to alleviate intracranial pressure related to multisutural craniosynostosis. The Chiari type I deformities in these patients were the biomechanical consequence of posterior fossa-cerebellar disproportion caused by supratentorial craniocerebral disproportion secondary to multisutural craniosynostosis. The authors postulate that all cases of Chiari type I deformity share the sine qua non feature of posterior fossa-cerebellar disproportion.


Long-term outcomes of patients with primary biliary cirrhosis and hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is an aggressive tumor that frequently develops in patients with primary biliary cirrhosis (PBC). We determined the mortality of patients with PBC who develop HCC, and which interventions (surgery, radiofrequency ablation, chemoembolization, alcohol injection, or transplantation) increase survival times. We investigated whether the Milan criteria predict outcomes of these patients and are effective in selection for liver transplantation. We evaluated data from 38 patients who had a confirmed diagnosis of PBC and HCC between March 1993 and February 2011. Patients were grouped based on whether or not they met the Milan criteria. Survival was assessed using the Kaplan-Meier analysis. Eighteen of the 38 patients (47.3%) died during the follow-up period; 49.4% survived for 5 years and 31.7% survived for 10 years. Thirty-five patients (92.0%) underwent one or a combination of interventions. Liver transplantation improved survival (risk ratio, 0.06; P < .0001), whereas surgery approached significance in causing deterioration (risk ratio, 2.87; P = .07). Mortality did not appear to be affected by meeting the Milan criteria (P = .84). Five- and 10-year survival times for patients with PBC who developed HCC were 49.4% and 31.7%, respectively. Patients who meet the Milan criteria receive liver transplantation as often as those who do not; we did not observe a difference in survival time between groups. Patients with PBC who develop HCC appear to benefit from aggressive therapies.


Communication between human mast cells and CD4(+) T cells through antigen-dependent interactions.
Mast cells (MCs) are immune cells residing in tissues where pathogens are first encountered. It has been indicated that MCs might also be involved in setting the outcome of T-cell responses. However, little is known about the capacity of human MCs to express MHC class II and/or to capture and present antigens to CD4(+) T cells. To study the T-cell stimulatory potential of human MCs, CD34(+) stem cell derived MCs were generated. These cells expressed HLA-DR when stimulated with IFN-γ, and, importantly, presented peptide and protein for activation of antigen-specific CD4(+) T cells. The interplay between MC and T cell led to increased HLA-DR expression on MCs. MCs were present in close proximity to T cells in tonsil and expressed HLA-DR and CD80, indicating their ability to present antigens to CD4(+) T cells in T-cell areas of human LNs. Our data show that MCs can present native antigens to human CD4(+) T cells and that HLA-DR expressing MCs are present in tonsil tissue, indicating that human MCs can directly activate T cells and provide a rationale to study the potential of MCs to prime and/or skew human T-cell responses.


Effects of oral contraceptive use on female sexual salivary hormones and indirect markers of muscle damage following eccentric cycling in women.
To determine the effects of oral contraceptive (OC) use on salivary concentrations of testosterone, estrogen, progesterone, and its effects on the changes in indirect markers of muscle damage following eccentric cycling in women. 10 oral contraceptive users at follicular phase (OC-FOL), 10 non-oral contraceptives users at follicular phase (NOC-FOL), and 10 non-oral contraceptives users at ovulation phase (NOC-OV) participated. Subjects performed 30 min of eccentric cycling at 90% of their maximal concentric power output (PO). Maximal voluntary isometric contraction (MVC), creatine kinase activity (CK), muscle soreness (SOR), and pain pressure threshold of vastus lateralis (PPT-VL) was assessed before, immediately after, and 24-96 h after cycling. Salivary estrogen, progesterone and testosterone concentrations were measured before, 72 and 96 h after exercise. No difference in estrogen levels between users and non-users was observed. Testosterone was 45% lower in OC-FOL than NOC-FOL at 96 h post-exercise (P = 0.01). Progesterone was 30.8-fold higher in NOC-OV than OC-FOL and 9.7-fold higher than NOC-FOL at 96 h post-exercise. The NOC-FOL recovered all indirect markers of muscle damage by 72 h post-exercise (P > 0.05). NOC-OV recovered MVC strength and muscle soreness (SOR and PPT-VL) by 96 h post-exercise (P > 0.05). OC-FOL did not recover baseline values of MVC, SOR, CK, and PPT-VL by 96 h. These results suggest that recovery after exercise-induced muscle damage took longer in OC-FOL, followed by NOC-OV and by NOC-FOL, respectively. Furthermore, testosterone and progesterone levels may affect recovery of indirect markers of muscle damage in women.


Multipoint targeting of TGF-β/Wnt transactivation circuit with microRNA 384-5p for cardiac fibrosis.
Cardiac fibrosis is a common precursor to ventricular dysfunction and eventual heart failure, and cardiac fibrosis begins with cardiac fibroblast activation. Here we have demonstrated that the TGF-β signaling pathway and Wnt signaling pathway formed a transactivation circuit during cardiac fibroblast activation and that miR-384-5p is a key regulator of the transactivation circuit. The results of in vitro study indicated that TGF-β activated an auto-positive feedback loop by increasing Wnt production in cardiac fibroblasts, and Wnt neutralizing antibodies disrupted the feedback loop. Also, we demonstrated that miR-384-5p simultaneously targeted the key receptors of the TGF-β/Wnt transactivation circuit and significantly attenuated both TGF-β-induced cardiac fibroblast activation and ischemia-reperfusion-induced cardiac fibrosis. In addition, small molecule that prevented pro-fibrogenic stimulus-induced downregulation of endogenous miR-384-5p significantly suppressed cardiac fibroblast activation and cardiac fibrosis. In conclusion, modulating a key endogenous miRNA targeting multiple components of the TGF-β/Wnt transactivation circuit can be an effective means to control cardiac fibrosis and has great therapeutic potential.


Watersoluble synthetic nucleic acid analogs--polyethyleneimine derivatives containing nucleic acid bases--conformation and interactionwith nucleic acids.
Water soluble polyethyleneimine derivatives containing nucleic acid bases were found to interact with polynucleotides, DNA, RNA. The conformational change by formation of complex was observed by CD spectra and was discussed with the hypochromicity in UV spectra. The rates of interactions between nucleic acid bases in polymers were slow as shown by UV spectra, but the conformational changes of the polynucleotides were fast as shown by CD spectra. In the case of the uracil derivative (PEI-Hse-Ura), high value of CD spectra [theta] 2.80 = -8.0 x 10(-4) for the complex with DNA might be caused by psi type conformation of DNA.


Augmentation of interleukin 1 and interleukin 2 production by OK-432.
Intraperitoneal (i.p.) administration of OK-432 augmented both interleukin 1 (IL-1) and interleukin 2 (IL-2) production to the rechallenge of OK-432 in vitro. Peritoneal exudate cells (PEC) of mice 8 days after i.p. injection with OK-432 (1 KE/mouse) showed maximum IL-1 production to the restimulation with OK-432 in vitro. OK-432-induced IL-1 was consisted of three molecular weight species (two major peaks: 85 K and 15 K daltons and one minor peak: 67 K daltons) on Sephadex G-100 chromatography. Splenocytes of mice 4 days after i.p. injection with OK-432 (1 KE/mouse) demonstrated maximum IL-2 production to the in vitro rechallenge of OK-432, however, in vivo OK-432 administration failed to enhance ConA-induced IL-2 production in vitro. From gel filtration analysis, OK-432 induced IL-2 had an unique molecular weight (approximately 70 K daltons). From these results, OK-432-induced augmentation of cellular immunity against tumor cells might be due to the activation of so-called lymphokine cascade reaction mediated by IL-1 and IL-2.


Analysis of corneal topographic and endothelial cell properties in newly diagnosed obstructive sleep apnea patients: A case-control study.
To investigate the topographic features and endothelial cell changes in the corneas of patients with newly diagnosed obstructive sleep apnea (OSA). This cross-sectional case-control study included 98 patients with OSA and 90 healthy controls. OSA was diagnosed on the basis of a polysomnographic sleep examination. Patients with OSA were grouped according to their apnea-hypopnea index (AHI) scores. Corneal topographic measurements were obtained using the Scheimpflug-Placido disk topographer, and keratometry values, anterior chamber parameters, thinnest corneal thickness, symmetry index (SI), keratoconus vertex (KV), and the Baiocchi-Calossi-Versaci index, including keratoconus (KC) screening indexes, were recorded. The corneal endothelium was visualizedby specular microscopy, and the endothelial cell density, coefficient of variability (CV), and percentage of hexagonal cells (HEX) were recorded. In this study, 32 patients had mild, 34 patients had moderate, and 32 patients had severe OSA. The mean back KV (KVb) and back SI values were significantly higher in the OSA group than in the control group (P <0.001, P = 0.035, respectively). As body mass index (BMI) increased, KVb also increased significantly (r = 0.281; P = 0.009). The mean CV and HEX values differed significantly between the OSA and control groups (P = 0.012 and P = 0.002, respectively). In addition, a significant correlation was found between AHI and KVb and CV values (r = 0.315; P <0.001 and r = 0.231; P = 0.011, respectively). This study revealed abnormal changes in the corneal endothelial morphology and topographic KC screening indexes in relation to OSA severity in patients and controls. Further research is required to determine the effect of BMI on corneal topographic findings in patients with OSA.


Pulmonary hypertension in an adult sickle cell population in Guadeloupe.
Pulmonary hypertension (PH) is a severe complication of sickle cell disease (SCD). The main purpose of this study was to evaluate the prevalence of PH in an adult SCD population in Guadeloupe. Echocardiography findings of 427 patients with SCD were analyzed. Subjects with a pulmonary artery systolic pressure (PASP) >30 mm Hg, at rest, were studied. PH was found in 49 (11.5%) SCD patients. Mean age was 37 years and mean PASP was 40 mm Hg. The most frequent symptom was exertional dyspnea. Thirteen subjects (26%) had left heart diseases. In the overall study population, 22 (5%) patients died. Seven (14%) patients had PH and 15 (4%) were PH free, p = 0.006. An early PH screening is recommended in SCD patients at steady state owing to the absence of specific symptoms and to the significant mortality rate related to this complication.


Cloning of a Lysobacter enzymogenes gene that encodes an arginyl endopeptidase (endoproteinase Arg-C).
Screening an expression library of Lysobacter enzymogenes DNA allowed us to clone a gene encoding a serine protease that cleaves synthetic substrates C-terminal to Arg and, to a lesser extent, Lys residues. The gene product, which shares sequence homology with the lysyl endopeptidases from L. enzymogenes and Achromobacter lyticus, consists of a signal sequence (24 residues), pro-region ( approximately 195 residues), and catalytic domain ( approximately 244 residues). Downstream of this gene is an open reading frame that lacks a promoter and appears to encode an inactive type I subtilase.


Live imaging reveals distinct modes of neutrophil and macrophage migration within interstitial tissues.
Cell motility is required for diverse processes during immunity and inflammation. Classically, leukocyte motility is defined as an amoeboid type of migration, however some leukocytes, like macrophages, also employ a more mesenchymal mode of migration. Here, we sought to characterize the mechanisms that regulate neutrophil and macrophage migration in vivo by using real-time imaging of leukocyte motility within interstitial tissues in zebrafish larvae. Neutrophils displayed a rounded morphology and rapid protease-independent motility, lacked defined paxillin puncta, and had persistent rearward polarization of stable F-actin and the microtubule network. By contrast, macrophages displayed an elongated morphology with reduced speed and increased directional persistence and formed paxillin-containing puncta but had a less-defined polarization of the microtubule and actin networks. We also observed differential effects of protease inhibition, microtubule disruption and ROCK inhibition on the efficiency of neutrophil and macrophage motility. Taken together, our findings suggest that larval zebrafish neutrophils and macrophage display distinct modes of migration within interstitial tissues in vivo.


Twin Pregnancies in Dairy Cattle: Observations in a Large Herd of Holstein-Friesian Dairy Cows.
Multiple pregnancies have devastating consequences on the herd economy of dairy cattle. This observational study examines incidence patterns based on data from the ultrasonographic examination of 1130 multiple pregnancies in cows in their third lactation or more carrying twins (98.8%), triplets (1.1%), or quadruplets (0.08%), and 3160 of their peers carrying singletons. Cows became pregnant following a spontaneous estrus with no previous hormone treatments. Irrespective of a significant decrease (p < 0.0001) in the conception rate (28-34 days post-insemination) during the warm period of the year, the multiple pregnancy rate was similar for both warm (26.5%) and cool (26.3%) periods. The incidence of unilateral multiple pregnancies (all embryos in the same uterine horn) was higher than that of bilateral pregnancies (at least one embryo in each uterine horn): 54.4% versus 45.6% (p < 0.0001). This difference rose to 17% during the warm season (p = 0.03). Pregnancy was monitored in unilateral multiple pregnancies until abortion or parturition (n = 615). In the warm period, the parturition rate was 43% compared to 61% recorded in the cool period (p < 0.0001). Thus, a warm climate is the main factor compromising the fate of multiple pregnancies. Some clinical suggestions are provided.


Immunostimulation exacerbates the biological effects of chemical carcinogens.
The reciprocal relation between the high regenerative ability of various animal species and the low incidence of haphazard or experimentally induced malignant tumours in these animal species is well documented. Equally well documented is the repeated observation that the decline in regenerative potential coincides with an increase in the incidence of cancers, a fact which, on an evolutionary scale, parallels with the development of a sophisticated immune system. The combination of the above observations led to the hypothesis that at least parts of an immune reaction might promote tumour development, and indeed, many experiments specifically designed to answer this question support this prediction. However, this "immunostimulation theory of tumour development" is neither explained in a satisfactory fashion nor universally adopted. The aim of the present investigation was to approach this issue by exploiting the dual, spectacular ability of urodele amphibians to regenerate a lot of organs and to make a stand to carcinogenesis. To this end, urodele amphibians of the species Triturus cristatus were immunologically challenged by intra-abdominal injections of sheep serum, they had then both their hind limbs amputated, and crystals of MNNG (N-Methyl-N"-nitro-N-nitrosoguanidine) were implanted into the stumps. The results show that the effects of MNNG on the immunostimulated animals display significant quantitative augmentation with respect to non-immunized controls. This augmentation consists in higher animal mortality, extension of the dedifferentiating stump tissue and concomitant retardation of limb restoration, and increase in the incidence of abnormal regenerates.


A Potential Association Between Retinal Changes, Subjective Memory Impairment, and Anxiety in Older Adults at Risk for Alzheimer's Disease: A 27-Month Pilot Study.
The utility of subjective memory impairment (SMI) as a risk marker for preclinical Alzheimer's disease (AD) remains unclear; however, recent studies have identified a correlation between retinal biomarkers and onset of preclinical disease. This study examines the relationship between retinal biomarkers that have been associated with cerebral amyloid, an early hallmark of AD, and SMI scores in patients at risk for developing AD. Forty-nine cognitively normal subjects were followed over 27 months and evaluated using a combination of neuropsychological, psychological, and retinal imaging instruments. Subjective memory testing was conducted using the memory assessment clinic questionnaire (MACQ) and Depression, Anxiety, and Stress Scales (DASS). Multivariate linear analysis was conducted using STATA software. Positive correlations were found between retinal nerve fiber layer (RNFL) volume and scores obtained from the MAC-Q at 27 months (MAC-Q_27), the DASS questionnaire for anxiety at 27 months (DASS-A_27), and the change in DASS-A over 27 months (dDASSA). There was also a significant positive correlation between these variables and the change in RNFL thickness over 27 months (dRNFL). MACQ_27, DASSA_27, and dDASS-A accounted for 35.7% of RFNL variance at 27 months and 21.5% of dRFNL variance. These findings suggest that worse subjective memory complaints and anxiety scores may be associated with one of the most commonly used structural anatomical retinal markers of early disease burden in AD. If so, these results lend support to SMI as a valid risk marker for later cognitive decline.


Antibiotic-lock technique is an effective treatment of bacterial catheter-related sepsis during parenteral nutrition.
Catheter-related sepsis is one of the most frequent and troublesome complications of parenteral nutrition. In a 2-year survey of 19 home parenteral nutrition patients, with a total of 25.2 years of cyclic nocturnal parenteral nutrition, the annual incidence of catheter-related sepsis was 1.27, of which 84% were due to bacterial catheter infection without any cutaneous focus. These 27 episodes were treated by a daily, 2 ml injection of antibiotic-saline solution, mainly amikacin, locked for 12 h per day within the infected catheter for 15 (7-20) days. On admission the parenteral nutrition was halted for 2 days and the catheter hub was changed. In 7 cases, an average of 3 days (2-5) of systemic antibiotic therapy was given in addition to the 2-week antibiotic-lock. Control of catheter-sepsis was achieved in 93% of the 27 episodes and parenteral nutrition was resumed using the same catheter with only one episode of recurrent sepsis. The present data confirm our preliminary report of the efficacy of the antibiotic-lock technique for the control of bacterial catheter-related sepsis. This treatment offers the advantage over current therapies of avoiding repeated catheter change and 2-6 weeks of systemic antibiotic therapy.


Role of the 4-hydroxy intermediate in the in vitro embryotoxicity of cyclophosphamide and dechlorocyclophosphamide.
Cyclophosphamide must be metabolically activated to produce malformations in cultured rat embryos. A 4-hydroxylated intermediate, 4-hydroxycyclophosphamide is initially formed during this activation. While 4-hydroxycyclophosphamide (and/or its open-ring tautomer, aldophosphamide) is believed to act as a transport form in mediating the antineoplastic activity of cyclophosphamide, its role in the teratogenicity of this drug is not known. In this study the effects of two "preactivated" cyclophosphamide analogs on cultured Day 10 rat embryos were determined. The first analog, 4-hydroperoxycyclophosphamide, is converted to 4-hydroxycyclophosphamide in aqueous solutions, releasing both acrolein and phosphoramide mustard, while the second, 4-hydroperoxydechlorocyclophosphamide, releases, in a similar manner, acrolein and the inactive metabolite, phosphoric acid diamide. Both cyclophosphamide analogs were teratogenic, embryolethal, and growth retarding in vitro, but the effective concentrations and the types of malformations produced were different. 4-Hydroperoxycyclophosphamide produced embryo deaths and malformations and decreases in embryonic growth and protein content at concentrations in the range of 5 to 25 microM. In contrast, 4-hydroperoxydechlorocyclophosphamide did not produce embryo deaths at concentrations below 100 microM and produced embryo malformations and growth retardation only at 125 microM. The concentration-response curve and the spectrum of malformations produced by 4-hydroperoxycyclophosphamide resembled those previously reported for phosphoramide mustard, while the concentration-response curve and types of malformations produced by 4-hydroperoxydechlorocyclophosphamide more closely resembled those observed with acrolein. Thus, the 4-hydroxy intermediates are similar as teratogens to the most potent of the metabolites which they produce; the 4-hydroxy compounds may serve as a transport form of cyclophosphamide but do not appear themselves to have a major role in teratogenicity.


Relationship between nomophobia and impulsivity among deaf and hard-of-hearing youth.
Excessive use of smartphones is known to be associated with negative social, physical, and psychological outcomes across age groups. A related problem is called "no-mobile-phone phobia" or "nomophobia," which is an extreme anxiety caused by not having access to a mobile phone. Despite their detrimental effects, smartphone use is highly prevalent among deaf/hard-of-hearing (DHH) individuals owing to their accessibility features. Therefore, it was deemed important to identify the prevalence of nomophobia in DHH youth and to examine the association between impulsivity and nomophobia. Gender-based differences in nomophobia and impulsivity were also examined. Data were collected from 187 DHH youth in Saudi Arabia using the Nomophobia Questionnaire and Barratt Impulsiveness Scale (short form). Findings revealed that 71.65% of the participants experienced severe nomophobia. While, nomophobia was more prevalent among female DHH youth than males, this difference was not observed for impulsivity. A linear regression analysis indicated that higher impulsivity was significantly associated with higher nomophobia in this sample. These findings suggest the importance of helping youth understand the disadvantages of smartphone use, and the consequences of their abuse or addiction to smartphones by incorporating this information into school curricula. Additionally, educating parents may help them monitor their children's smartphone use more closely.


Production of Aspergillus terreus alpha-L-rhamnosidase by solid state fermentation.
The study of production of Aspergillus terreus CECT 2663 alpha-L-rhamnosidase in solid state fermentation using wheat bran, washed sugar cane bagasse and polyurethane foam as substrates or supports for the enzyme production. Cultures were carried out in Petri dishes under controlled temperature and humidity. Naringin or rhamnose were the enzyme inducers and carbon sources. The enzyme activity to inducer ratio was appreciably greater when using sugar cane bagasse or polyurethane foam than wheat bran. The influence of inoculum size, inducer, airflow, humidity and temperature were determined. Under optimum conditions, about four units of enzyme per ml nutrient solution were obtained after 4-6 d. The activity to inducer ratio was higher, and the cultivation time was shorter in solid state fermentation than those observed in submerged cultures. Solid cultures, using naringin as inducer, can be appropriate alpha-L-rhamnosidase production.


Fire regime of peatlands in the Angolan Highlands.
The Angolan Highlands region includes the Angolan miombo woodland ecoregion which supports miombo woodland, grasslands, subsistence agricultural land, and peatland deposits. Extensive fires, slash and burn agriculture, peat fuel extraction, and peatland drainage are among the anthropogenic practices that threaten these peatland deposits. Peat fires cause peatland degradation, release significant amounts of greenhouse gases, deteriorate air quality, and contribute towards climate change and biodiversity loss. This study presents an analysis of the fire regimes over the period 2001 to 2020 in an under-studied area of the Angolan Highlands. Moderate Resolution Imaging Spectroradiometer (MODIS) fire and vegetation data were used in combination with a land use/land cover (LULC) classification map to calculate fire frequency, burn area, and fire regimes. The fire patterns within the study site are comparable to those found in African woodland savannas. Across the study site, 6976 km2 (11.31%) of the land surface area burned at least nine times from 2001 to 2020, occurring largely within in the river valley environment. Considering the different LULC classes, peatlands were calculated to (a) burn more frequently (average fire frequency from 2001 to 2020 = 9.12), (b) have the smallest proportion (4.11%) of area which remained unburnt over the fire archive, and (c) have the largest average proportion (45.65% or 746 km2) of burnt area per year. Peatland burning occurred predominantly during drier months from May to September. The results of this study highlight the strong influence of LULC on the fire frequency and distribution in the study area, requiring unique fire management strategies. As has been documented for boreal and tropical peatlands across the globe, we stress the importance of peatland conservation and protection; continued unsustainable management practices may lead to the loss of these important peatland deposits.


Urinothorax as a manifestation of nondilated obstructive uropathy following renal transplantation.
A 12-year-old patient developed prolonged nondilated urinary obstruction and pleural effusion shortly after undergoing renal transplantation. Renal sonography, angiography, and isotope renography failed to identify an obstructive process. On the 18th postoperative day, pleural effusion was noted in the right hemithorax, and by day 24, increased perinephric fluid was observed on renal scan. Following a nephrostomy, the pleural effusion resolved and renal function improved remarkably. A ureterovesical junction obstruction and renal pelvis tear that were later discovered were repaired. Whenever a ureteral obstruction is suspected the diagnosis should be pursued vigorously, despite normal radiologic findings, especially in the presence of pleural effusion. Consideration of the possibility of urinothorax in such cases may obviate the need for lung biopsy.


Metabolic and hemodynamic effects of high-dose insulin treatment in aortic valve and coronary surgery.
Glucose and insulin have been used as an adjuvant therapy in cardiac surgery because of their potentially beneficial effects on myocardial metabolism and contractile function. This study evaluated the effects of high-dose insulin on systemic metabolism and hemodynamics after combined heart surgery. Forty elective patients scheduled for combined aortic valve replacement and coronary artery bypass surgery were randomly assigned to receive either high-dose insulin treatment (short-acting insulin 1 IU.kg(-1).h(-1) with 30% glucose 1.5 mL.kg(-1).h(-1) administered separately) or control treatment (saline). The blood glucose levels were maintained within a targeted range by adjusting the rate of glucose infusion in the treatment group and by short-acting insulin bolus doses in the control group. The lactate clearance was faster (p = 0.046), and the lactate levels (p = 0.016), blood glucose levels (p < 0.001), and free fatty acid levels (p < 0.001) were lower in the insulin group postoperatively. Besides, there was lesser need for dobutamine support (p = 0.013) and a trend toward better cardiac indices. Insulin treatment increased the respiratory quotient (p < 0.001), but there were no differences between the groups with regard to systemic oxygen consumption or energy expenditure measured by indirect calorimetry. The average glucose uptake in the insulin group was 7.1 g/kg in 24 hours (28 kcal.kg(-1).day(-1)). The high-dose insulin treatment was associated with lower blood glucose levels, better preserved myocardial contractile function, and less need for inotropic support, and hence led to lower lactate levels postoperatively. The protocol is safe, but requires strict control of blood glucose level.


Purification and characterization of (Na+ + K+)-ATPase from toad kidney.
This report describes the partial purification and the characteristics of (Na+ + K+)-ATPase (ATP phosphohydrolase, EC 188.8.131.52) from an amphibian source. Toad kidney microsomes were solubilized with sodium deoxycholate and further purified by sodium dodecyl sulphate treatment and sucrose gradient centrifugation, according to the methods described by Lane et al. [(1973) J. Biol. Chem. 248, 7197--7200], Jørgensen [(1974) Biochim. Biophys. Acta 356, 36--52] and Hayashi et al. [(1977) Biochim. Biophys. Acta 482, 185--196]. (Na+ + K+)-ATPase preparations with specific activities up to 1000 mumol Pi/mg protein per h were obtained. Mg2+-ATPase only accounted for about 2% of the total ATPase activity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed three major protein bands with molecular weights of 116 000, 62 000 and 26 000. The 116 000 dalton protein was phosphorylated by [gamma-32P]ATP in the presence of sodium but not in the presence of potassium. The 62 000 dalton component stained for glycoproteins. The Km for ATP was 0.40 mM, for Na+ 12.29 mM and for K+ 1.14 mM. The Ki for ouabain was 35 micron. Temperature activation curves showed two activity peaks at 37 degrees C and at 50 degrees C. The break in the Arrhenius plot of activity versus temperature appeared at 15 degrees C.


The effect of dexmedetomidine on inflammatory response of septic rats.
Some studies have demonstrated dexmedetomidine has anti-inflammatory effect on septic rats. However, the mechanism of how dexmedetomidine exerts these effects is still remained unknown. This study was designed to investigate the mechanism of how dexmedetomidine inhibits the production of inflammatory mediators in cecal ligation and puncturinduced septic rats. 48 Sprague-Dawley rats were randomly divided into six groups: sham-operated (sham) group, cecal ligation and puncture (CLP) group, dexmedetomidine 5 μg/kg (DEX5) group, dexmedetomidine 10 μg/kg (DEX10) group,dexmedetomidine + yohimbine (DEX10 + Yoh) group and yohimibine group (Yoh). Blood, bronchoalveolarlavage fluid (BALF) and lung tissues in each group were collected at six hours after dexmedetomidine or yohimbine treatment,. Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in BALF and plasma were measured by enzyme-linked immunosorbent assay (ELISA). Toll-like receptor-4(TLR4) and myeloid differerntiation factor(MyD88) expression were measuredby quantitative PCR, and extracellular signal-regulated kinase (ERK) 1/2 phosphorylation were determined by western blott. Compared with CLP group, dexmedetomidine significantly decreased not only the production of TNF-α and IL-6 both in plasma and BALF, but also inhibited the expression of TLR4 and MyD88 in mRNA level and the activation of ERK1/2 and NF-κB in the lung tissues of CLP-induced septic rats. All these effects could not be reversed by yohimibine. Dexmedetomidine treatment can effectively reduce the generation of inflammatory mediators in the plasma and BALF of CLP-induced septic rats. These effects of dexmedetomidine rely on TLR4/MyD88/MAPK/ NF-κB signaling pathway and are independent of α2-adrenoceptor.


Collaborative science in the next-generation sequencing era: a viewpoint on how to combine exome sequencing data across sites to identify novel disease susceptibility genes.
The purpose of this article is to inform readers about technical challenges that we encountered when assembling exome sequencing data from the 'Simplifying Complex Exomes' (SIMPLEXO) consortium-whose mandate is the discovery of novel genes predisposing to breast and ovarian cancers. Our motivation is to share these obstacles-and our solutions to them-as a means of communicating important technical details that should be discussed early in projects involving massively parallel sequencing.


New observations on minifascicular neuropathy with sex-dependent gonadal dysgenesis: a case series with nerve ultrasound assessment.
Gonadal dysgenesis with minifascicular neuropathy (GDMN) is a rare autosomal recessive condition associated with biallelic DHH pathogenic variants. In 46, XY individuals, this disorder is characterized by an association of minifascicular neuropathy (MFN) and gonadal dysgenesis, while in 46, XX subjects only the neuropathic phenotype is present. Very few patients with GDMN have been reported so far. We describe four patients with MFN due to a novel DHH likely pathogenic homozygous variant and the results of nerve ultrasound assessment. This retrospective observational study included 4 individuals from 2 unrelated Brazilian families evaluated for severe peripheral neuropathy. Genetic diagnosis was performed with a peripheral neuropathy next-generation sequencing (NGS) panel based on whole exome sequencing focused analysis that included a control SRY probe to confirm genetic sex. Clinical characterization, nerve conduction velocity studies, and high-resolution ultrasound nerve evaluation were performed in all subjects. Molecular analysis disclosed in all subjects the homozygous DHH variant p.(Leu335Pro). Patients had a striking phenotype, with marked trophic changes of extremities, sensory ataxia, and distal anesthesia due to a sensory-motor demyelinating polyneuropathy. One 46, XY phenotypically female individual had gonadal dysgenesis. High-resolution nerve ultrasound showed typical minifascicular formation and increased nerve area in at least one of the nerves assessed in all patients. Gonadal dysgenesis with minifascicular neuropathy is a severe autosomal recessive neuropathy characterized by trophic alterations in limbs, sensory ataxia, and distal anesthesia. Nerve ultrasound studies are very suggestive of this condition and may help to avoid invasive nerve biopsies.


Single-stage extensive replacement of the thoracic aorta: the arch-first technique.
Single-stage extensive replacement of the thoracic aorta usually involves a period of circulatory arrest with performance of the graft-to-lower descending thoracic aorta anastomosis before performing the anastomosis to the arch vessels. To minimize the period of brain ischemia and reduce the potential for neurologic injury, we developed an alternative technique. In 6 patients with extensive aneurysms involving the entire thoracic aorta, exposure was obtained via a bilateral thoracotomy in the anterior fourth intercostal space with transverse sternotomy. A 10-mm graft was anastomosed to the aortic graft, opposite the site of the planned anastomosis to the arch vessels. During a single period of circulatory arrest (34-46 minutes), the aortic graft was attached to a cuff of aorta containing the arch vessels. The graft was then clamped on either side, and the arch was perfused with cold blood for 20 to 36 minutes. After the distal aortic anastomosis was completed, antegrade perfusion was established via the 10-mm graft. The proximal aortic anastomosis was performed last. No patient sustained a permanent neurologic deficit. All 6 patients were discharged from the hospital. The "arch-first" technique, combined with a bilateral transverse thoracotomy, allows expeditious replacement of the thoracic aorta with an acceptable interval of hypothermic circulatory arrest and minimizes the risk of retrograde atheroembolism by establishing antegrade perfusion.


Amodiaquine fails to cure chloroquine resistant Plasmodium falciparum in the Punjab.
We evaluated amodiaquine as a replacement drug for treating falciparum malaria in an area of Pakistani Punjab where chloroquine-resistant Plasmodium falciparum has recently emerged. Amodiaquine appeared to be 4 to 8 times more effective than chloroquine when P. falciparum isolates were tested in vitro. However, the recrudescence rate was greater than 50% after oral treatment with 20 mg/kg amodiaquine given over two successive days. This lack of therapeutic response from amodiaquine may have been due to selection of resistant parasites in the villages where the study was performed through extensive use of chloroquine for presumptive malaria treatment during the preceding 18 months. We conclude that amodiaquine is not a suitable replacement for chloroquine for treating falciparum malaria in our study area despite in vitro sensitivity data suggesting that it would be efficacious. Baseline in vitro sensitivity to mefloquine is also reported.


Depression care management for adults older than 60 years in primary care clinics in urban China: a cluster-randomised trial.
China's national health policy classifies depression as a chronic disease that should be managed in primary care settings. In some high-income countries use of chronic disease management principles and primary care-based collaborative-care models have improved outcomes for late-life depression; however, this approach has not yet been tested in China. We aimed to assess whether use of a collaborative-care depression care management (DCM) intervention could improve outcomes for Chinese adults with depression aged 60 years and older. Between Jan 17, 2011, [corrected] and Nov 30, 2013, we did a cluster-randomised trial in patients from primary care centre clinics in Shangcheng district of Hangzhou city in eastern China. We randomly assigned (1:1) clinics to either DCM (involving training for physicians in use of treatment guidelines, training for primary care nurses to function as care managers, and consultation with psychiatrists as support) or to give enhanced care as usual to all eligible patients aged 60 years and older with major depressive disorder. Clinics were chosen randomly for inclusion from all primary care clinics in the district by computer algorithm and then randomly allocated depression care interventions remotely by computer algorithm. Physicians, study personnel, and patients were not masked to clinic assignment. Our primary outcome was difference in Hamilton Depression Rating Scale (HAMD) score using data for clusters at baseline and 3, 6, and 12 month follow-up in a mixed-effects model of the intention-to-treat population. We originally aimed to analyse outcomes at 24 months, however the difference between groups at 12 months was large and funding was insufficient to continue to 24 months, therefore we decided to end the trial at 12 months. This trial is registered with ClinicalTrials.gov, number NCT01287494. Of 34 primary care clinics in Shangcheng district, 16 were randomly chosen. We randomly assigned eight clinics to the DCM intervention (164 patients enrolled) and eight primary care clinics to enhanced care as usual (162 patients). There were no major differences in baseline demographic and clinical variables between the groups of patients for each intervention. Over the 12 months, patients in clinics assigned to DCM had a significantly greater reduction in HAMD score than did those in practices assigned to enhanced care as usual (estimated between group difference -6·5 [95% CI -7·1 to -5·9]; Cohen's d 0·8 [95% CI 0·8-0·9]; p<0·0001). The intercluster correlation for change in HAMD total score was 0·07 (95% CI 0·06-0·08). There were no study-related adverse events in either group. Clinical outcomes of Chinese adults older than 60 years who had major depression were improved when their primary care clinic used DCM. Primary care-based collaborative management of depression is promising to address this pressing public health need in China. National Institutes of Health, Program for New Century Excellent Talents in Universities of China, Ministry of Education, China.


A genomic-scale artificial microRNA library as a tool to investigate the functionally redundant gene space in Arabidopsis.
Traditional forward genetic screens are limited in the identification of homologous genes with overlapping functions. Here, we report the analyses and assembly of genome-wide protein family definitions that comprise the largest estimate for the potentially redundant gene space in Arabidopsis thaliana. On this basis, a computational design of genome-wide family-specific artificial microRNAs (amiRNAs) was performed using high-performance computing resources. The amiRNA designs are searchable online (http://phantomdb.ucsd.edu). A computationally derived library of 22,000 amiRNAs was synthesized in 10 sublibraries of 1505 to 4082 amiRNAs, each targeting defined functional protein classes. For example, 2964 amiRNAs target annotated DNA and RNA binding protein families and 1777 target transporter proteins, and another sublibrary targets proteins of unknown function. To evaluate the potential of an amiRNA-based screen, we tested 122 amiRNAs targeting transcription factor, protein kinase, and protein phosphatase families. Several amiRNA lines showed morphological phenotypes, either comparable to known phenotypes of single and double/triple mutants or caused by overexpression of microRNAs. Moreover, novel morphological and abscisic acid-insensitive seed germination mutants were identified for amiRNAs targeting zinc finger homeodomain transcription factors and mitogen-activated protein kinase kinase kinases, respectively. These resources provide an approach for genome-wide genetic screens of the functionally redundant gene space in Arabidopsis.


Quantitative evaluation of hemodynamic response after hypercapnia among different brain territories by fMRI.
The brain vascular system has an autoregulatory mechanism that maintains blood perfusion within normal limits at the capillary level. Partially due to its clinical importance, it is of interest to better understand the mechanisms involved in vascular regulation. Therefore, using functional magnetic resonance imaging (fMRI), we quantitatively investigated hemodynamic response characteristics of regions supplied by the main cerebral arteries, during two breath holding tests (BHT): after inspiration and after expiration. We used an auto-regressive method capable of estimating four signal parameters: onset delay, full width at half maximum (FWHM), time-to-peak and amplitude. The onset delay was significantly longer for the posterior cerebral artery (PCA) than for middle cerebral artery (MCA) and anterior arteries (ACA). FWHM and time-to-peak were larger in the ACA territory, indicating a slower blood flow in this region. Differences were also observed in the amplitude among the three areas, where MCA and PCA territories showed the smallest and the highest amplitudes, respectively. Moreover, differences were found in amplitude and onset when BHT was performed after inspiration as compared to BHT after expiration. Time-to-peak and FWHM showed no statistical differences between these two challenges. Such results are related to regional anatomical specificities and biochemical mechanisms responsible for vasodilation, such as those related to vascularity and vessel sizes.


Delayed brain metastasis in recurrent hepatocellular carcinoma following liver transplantation: a case report highlighting the predictive value of microvascular invasion.
Recurrent hepatocellular carcinoma (HCC) poses a significant challenge after liver transplantation, affecting approximately 10-23% of patients with a median onset of 13 months post-transplantation. Extrahepatic involvement, such as lung, bone, adrenal glands, peritoneum, lymph nodes, and central nervous system (CNS), is commonly observed among transplant recipients with HCC recurrence. Notably, vascular invasion (VI), including microvascular invasion (MiVI) and macrovascular invasion (MVI), substantially increase the risk of recurrence by 2.42- and 7.82-fold, respectively. This article presents a unique case of a 72-year-old male patient with a history of HCV-related cirrhosis and HCC who underwent orthotopic liver transplantation (OLT). Six years later, he presented to the emergency department following a fall, which led to the discovery of a pathologic fracture of T7 and an incidental intracranial mass during imaging. Subsequent biopsy confirmed metastatic HCC in the T7 lesion, while magnetic resonance imaging revealed two enhancing brain masses. One mass measured 4.8 cm in the left occipitotemporal lobe, and the other measured 1.7 cm in the right frontal gyrus. Notably, the patient had exhibited MiVI and a mildly elevated alpha-fetoprotein level (AFP) of 7.6 ng/mL at the time of his OLT. This case underscores the predictive value of MiVI in HCC recurrence post-OLT. Accordingly, extended post-transplantation surveillance is crucial for patients with HCC and MiVI. Moreover, this report highlights the uncommon occurrence of delayed brain metastasis following OLT in a patient with HCC.


Altered Vitamin D receptor-coactivator interactions reflect superagonism of Vitamin D analogs.
The active form of Vitamin D, 1alpha,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)], has potent antiproliferative actions on various normal and malignant cells. Calcemic effects, however, hamper therapeutic application of 1,25-(OH)(2)D(3) in hyperproliferative diseases. Two 14-epi-analogs of 1,25-(OH)(2)D(3) namely 19-nor-14-epi-23-yne-1,25-(OH)(2)D(3) (TX522) and 19-nor-14,20-bisepi-23-yne-1,25-(OH)(2)D(3) (TX527), display reduced calcemic effects coupled to an (at least 10-fold) increased antiproliferative potency when compared with 1,25-(OH)(2)D(3). Altered cofactor recruitment by the Vitamin D receptor (VDR) might underlie the superagonism of these 14-epi-analogs. Therefore, this study aims to evaluate their effects at the level of VDR-coactivator interactions. Mammalian two-hybrid assays with VDR and the coactivators TIF2 and DRIP205 showed the 14-epi-analogs to be more potent inducers of VDR-coactivator interactions than 1,25-(OH)(2)D(3). TX522 and TX527 require 30- and 40-fold lower doses to obtain the VDR-DRIP205 interaction induced by 1,25-(OH)(2)D(3) at 10(-8)M. Evaluation of additional 1,25-(OH)(2)D(3)-analogs and their impact on VDR-coactivator interactions revealed a strong correlation between the antiproliferative potency of an analog and its ability to induce VDR-coactivator interactions. In conclusion, these data show that altered coactivator binding by the VDR is one possible explanation for the superagonistic action of the two 14-epi-analogs TX522 and TX527.


Cell tracking using phase-adaptive shape prior.
Automated tracking of cell population is very crucial for quantitative measurements of dynamic cell-cycle behaviour of individual cells. This problem involves several subproblems and a high accuracy of each step is essential to avoid error propagation. In this paper, we propose a holistic three-component system to tackle this problem. For each phase, we first learn a mean shape as well as a model of the temporal dynamics of transformation, which are used for estimating a shape prior for the cell in the current frame. We then segment the cell using a level set-based shape prior model. Finally, we identify its phase based on the goodness-of-fit of the data to the segmentation model. This phase information is also used for fine-tuning the segmentation result. We evaluate the performance of our method empirically in various aspects and in tracking individual cells from HeLa H2B-GFP cell population. Highly accurate validation results confirm the robustness of our method in many realistic scenarios and the essentiality of each component of our integrating system.


V1 neurons respond to luminance changes faster than contrast changes.
Luminance and contrast are two major attributes of objects in the visual scene. Luminance and contrast information received by visual neurons are often updated simultaneously. We examined the temporal response properties of neurons in the primary visual cortex (V1) to stimuli whose luminance and contrast were simultaneously changed by 50 Hz. We found that response tuning to luminance changes precedes tuning to contrast changes in V1. For most V1 neurons, the onset time of response tuning to luminance changes was shorter than that to contrast changes. Most neurons carried luminance information in the early response stage, while all neurons carried both contrast and luminance information in the late response stage. The early luminance response suggests that cortical processing for luminance is not as slow as previously thought.


Genome-wide transcriptional variation within and between steady states for continuous growth of the hyperthermophile Thermotoga Maritima.
Maltose-limited, continuous growth of the hyperthermophile Thermotoga maritima at different temperatures and dilution rates (80 degrees C/0.25 h(-1), 80 degrees C/0.17 h(-1), and 85 degrees C/0.25 h(-1)) showed that transcriptome-wide variation in gene expression within mechanical steady states was minimal compared to that between steady states, supporting the efficacy of chemostat-based approaches for functional genomics studies.


Cost analysis for efficient management: diabetes treatment at a public district hospital in Thailand.
OBJECTIVE The study estimated cost of illness from the provider's perspective for diabetic patients who received treatment during the fiscal year 2008 at Waritchaphum Hospital, a 30-bed public district hospital in Sakhon Nakhon province in northeastern Thailand. METHODS This retrospective, prevalence-based cost-of-illness study looked at 475 randomly selected diabetic patients, identified by the World Health Organization's International Classification of Diseases, 10th revision, codes E10-E14. Data were collected from the hospital financial records and medical records of each participant and were analysed with a stepwise multiple regression. KEY FINDINGS The study found that the average public treatment cost per patient per year was US$94.71 at 2008 prices. Drug cost was the highest cost component (25% of total cost), followed by inpatient cost (24%) and outpatient visit cost (17%). A cost forecasting model showed that length of stay, hospitalization, visits to the provincial hospital, duration of disease and presence of diabetic complications (e.g. diabetic foot complications and nephropathy) were the significant predictor variables (adjusted R(2) = 0.689). CONCLUSIONS According to the fitted model, avoiding nephropathy and foot complications would save US$19 386 and US$39 134 respectively per year. However, these savings are missed savings for the study year and the study hospital only and not projected savings, as that would depend on the number of diabetic patients managed in the year, the ratio of complicated to non-complicated cases and effectiveness of the prevention programmes. Nonetheless, given the high avoidable cost associated with complications of diabetes, healthcare providers in Thailand should focus on initiatives that delay the progression of complications in diabetic patients.