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Dosing in neonates: Special considerations in physiology and trial design. |
Determining the right dose for drugs used to treat neonates is critically important. Neonates have significant differences in physiology affecting drug absorption, distribution, metabolism, and elimination that makes extrapolating dosages from adults and older children inappropriate. In spite of recent legislative efforts requiring drug studies in this population, most drugs given to neonates remain insufficiently studied. Many ethical and logistical concerns make designing studies in this age group difficult. Fortunately, specialized analytical techniques, such as the use of dried blood spots, scavenged sampling, population pharmacokinetics analyses, and sparse sampling, have helped investigators better define doses that maximize efficacy and safety. Through the use of these methods, successful clinical trials have resulted in recent changes to drug dosing in this population.Pediatric Research (2014); doi:10.1038/pr.2014.143. |
Diabetes-associated changes in the fetal insulin/insulin-like growth factor system are organ specific in rats. |
Background:Diabetes in pregnancy affects fetal growth and development. The insulin/IGF system comprising insulin, insulin-like growth factors, their receptors and binding proteins, has been implicated in fetal growth regulation. This study tested the hypothesis that maternal diabetes alters the fetal insulin/IGF system in a tissue specific manner.Methods:Wistar rats were rendered diabetic by neonatal administration of streptozotocin and mated with control rats. At day 21 of gestation the weights of fetuses, placentas and fetal organs (heart, lung, liver, stomach, intestine and pancreas) were determined. Maternal and fetal plasma concentrations of insulin, IGF1 and IGF2 were measured by ELISA, and expression of IGF1, IGF2, IGF1R, IGF2R, IR, IGFBP1, BP2 and BP3 in placenta and fetal organs by qPCR.Results:The well known increase in fetal growth in this model of mild diabetes is accompanied by elevated insulin and IGF1 levels and alterations of the insulin/IGF system in the fetus and the placenta. These alterations were organ and gene specific. The insulin/IGF system was generally upregulated, especially in the fetal heart, whilst it was downregulated in fetal lung.Conclusion:In our model of mild diabetes the effect of maternal diabetes on fetal weight and fetal insulin/IGF system expression is organ specific with highly sensitive organs such as lung and heart, and organs that were less affected, such as stomach.Pediatric Research (2014); doi:10.1038/pr.2014.139. |
Abnormal Calcium Handling and Exaggerated Cardiac Dysfunction in Mice with Defective Vitamin D Signaling. |
AIM: Altered vitamin D signaling is associated with cardiac dysfunction, but the pathogenic mechanism is not clearly understood. We examine the mechanism and the role of vitamin D signaling in the development of cardiac dysfunction. METHODS AND RESULTS: We analyzed 1alpha-hydroxylase (1alpha-OHase) knockout (1alpha-OHase-/-) mice, which lack 1alpha-OH enzymes that convert the inactive form to hormonally active form of vitamin D. 1alpha-OHase-/- mice showed modest cardiac hypertrophy at baseline. Induction of pressure overload by transverse aortic constriction (TAC) demonstrated exaggerated cardiac dysfunction in 1alpha-OHase-/- mice compared to their WT littermates with a significant increase in fibrosis and expression of inflammatory cytokines. Analysis of calcium (Ca2+) transient demonstrated profound Ca2+ handling abnormalities in 1alpha-OHase-/- mouse cardiomyocytes (CMs), and treatment with paricalcitol (PC), an activated vitamin D3 analog, significantly attenuated defective Ca2+ handling in 1alpha-OHase-/- CMs. We further delineated the effect of vitamin D deficiency condition to TAC by first correcting the vitamin D deficiency in 1alpha-OHase-/- mice, followed then by either a daily maintenance dose of vitamin D or vehicle (to achieve vitamin D deficiency) at the time of sham or TAC. In mice treated with vitamin D, there was a significant attenuation of TAC-induced cardiac hypertrophy, interstitial fibrosis, inflammatory markers, Ca2+ handling abnormalities and cardiac function compared to the vehicle treated animals. CONCLUSIONS: Our results provide insight into the mechanism of cardiac dysfunction, which is associated with severely defective Ca2+ handling and defective vitamin D signaling in 1alpha-OHase-/- mice. |
Whole Exome Analysis Identifies Frequent CNGA1 Mutations in Japanese Population with Autosomal Recessive Retinitis Pigmentosa. |
OBJECTIVE: The purpose of this study was to investigate frequent disease-causing gene mutations in autosomal recessive retinitis pigmentosa (arRP) in the Japanese population. METHODS: In total, 99 Japanese patients with non-syndromic and unrelated arRP or sporadic RP (spRP) were recruited in this study and ophthalmic examinations were conducted for the diagnosis of RP. Among these patients, whole exome sequencing analysis of 30 RP patients and direct sequencing screening of all CNGA1 exons of the other 69 RP patients were performed. RESULTS: Whole exome sequencing of 30 arRP/spRP patients identified disease-causing gene mutations of CNGA1 (four patients), EYS (three patients) and SAG (one patient) in eight patients and potential disease-causing gene variants of USH2A (two patients), EYS (one patient), TULP1 (one patient) and C2orf71 (one patient) in five patients. Screening of an additional 69 arRP/spRP patients for the CNGA1 gene mutation revealed one patient with a homozygous mutation. CONCLUSIONS: This is the first identification of CNGA1 mutations in arRP Japanese patients. The frequency of CNGA1 gene mutation was 5.1% (5/99 patients). CNGA1 mutations are one of the most frequent arRP-causing mutations in Japanese patients. |
Aurora Kinases as Targets in Drug-Resistant Neuroblastoma Cells. |
Aurora kinase inhibitors displayed activity in pre-clinical neuroblastoma models. Here, we studied the effects of the pan-aurora kinase inhibitor tozasertib (VX680, MK-0457) and the aurora kinase inhibitor alisertib (MLN8237) that shows some specificity for aurora kinase A over aurora kinase B in a panel of neuroblastoma cell lines with acquired drug resistance. Both compounds displayed anti-neuroblastoma activity in the nanomolar range. The anti-neuroblastoma mechanism included inhibition of aurora kinase signalling as indicated by decreased phosphorylation of the aurora kinase substrate histone H3, cell cycle inhibition in G2/M phase, and induction of apoptosis. The activity of alisertib but not of tozasertib was affected by ABCB1 expression. Aurora kinase inhibitors induced a p53 response and their activity was enhanced in combination with the MDM2 inhibitor and p53 activator nutlin-3 in p53 wild-type cells. In conclusion, aurora kinases are potential drug targets in therapy-refractory neuroblastoma, in particular for the vast majority of p53 wild-type cases. |
Urotensin II Inhibits Doxorubicin-Induced Human Umbilical Vein Endothelial Cell Death by Modulating ATF Expression and via the ERK and Akt Pathway. |
BACKGROUND AND PURPOSE: Regulation of the homeostasis of vascular endothelium is critical for the processes of vascular remodeling and angiogenesis under physiological and pathological conditions. Urotensin II (U-II), a potent vasoactive peptide, participates in vascular and myocardial remodeling after injury. We investigated the protective effect of U-II on doxorubicin (DOX)-induced apoptosis in cultured human umbilical vein endothelial cells (HUVECs) and the potential mechanisms involved in this process. EXPERIMENTAL APPROACH: Cultured HUVECs were treated with vehicle, DOX (1 microM), U-II, or U-II plus DOX. Apoptosis was evaluated by DNA strand break level with TdT-mediated dUTP nick-end labeling (TUNEL) staining. Western blot analysis was employed to determine the related protein expression and flow cytometry assay was used to determine the TUNEL positive cells. KEY RESULTS: U-II reduced the quantity of cleaved caspase-3 and cytosol cytochrome c and increased Bcl-2 expression, which results in protecting HUVECs from DOX-induced apoptosis. U-II induced Activating transcription factor 3 (ATF3) at both mRNA and protein levels in U-II-treated cells. Knockdown of ATF3 with ATF3 siRNA significantly reduced ATF3 protein levels and U-II protective effect under DOX-treated condition. U-II downregulated p53 expression in DOX-induced HUVECs apoptosis, and it rapidly activated extracellular signal-regulated protein kinase (ERK) and Akt. The DOX induced change of p53 was not affected by U-II antagonist (urantide) under ATF-3 knockdown. The inhibitory effect of U-II on DOX-increased apoptosis was attenuated by inhibitors of ERK (U0126) and PI3K/Akt (LY294002). CONCLUSION AND IMPLICATIONS: Our observations provide evidence that U-II protects HUVECs from DOX-induced apoptosis. ERK-Akt phosphorylation, ATF3 activation, and p53 downregulation may play a signal-transduction role in this process. |
The Indolinone MAZ51 Induces Cell Rounding and G2/M Cell Cycle Arrest in Glioma Cells without the Inhibition of VEGFR-3 Phosphorylation: Involvement of the RhoA and Akt/GSK3beta Signaling Pathways. |
MAZ51 is an indolinone-based molecule originally synthesized as a selective inhibitor of vascular endothelial growth factor receptor (VEGFR)-3 tyrosine kinase. This study shows that exposure of two glioma cell lines, rat C6 and human U251MG, to MAZ51 caused dramatic shape changes, including the retraction of cellular protrusions and cell rounding. These changes were caused by the clustering and aggregation of actin filaments and microtubules. MAZ51 also induced G2/M phase cell cycle arrest. This led to an inhibition of cellular proliferation, without triggering significant cell death. These alterations induced by MAZ51 occurred with similar dose- and time-dependent patterns. Treatment of glioma cells with MAZ51 resulted in increased levels of phosphorylated GSK3beta through the activation of Akt, as well as increased levels of active RhoA. Interestingly, MAZ51 did not affect the morphology and cell cycle patterns of rat primary cortical astrocytes, suggesting it selectively targeted transformed cells. Immunoprecipitation-western blot analyses indicated that MAZ51 did not decrease, but rather increased, tyrosine phosphorylation of VEGFR-3. To confirm this unanticipated result, several additional experiments were conducted. Enhancing VEGFR-3 phosphorylation by treatment of glioma cells with VEGF-C affected neither cytoskeleton arrangements nor cell cycle patterns. In addition, the knockdown of VEGFR-3 in glioma cells did not cause morphological or cytoskeletal alterations. Furthermore, treatment of VEGFR-3-silenced cells with MAZ51 caused the same alterations of cell shape and cytoskeletal arrangements as that observed in control cells. These data indicate that MAZ51 causes cytoskeletal alterations and G2/M cell cycle arrest in glioma cells. These effects are mediated through phosphorylation of Akt/GSK3beta and activation of RhoA. The anti-proliferative activity of MAZ51 does not require the inhibition of VEGFR-3 phosphorylation, suggesting that it is a potential candidate for further clinical investigation for treatment of gliomas, although the precise mechanism(s) underlying its effects remain to be determined. |
Readmissions after Hospitalization for Heart Failure, Acute Myocardial Infarction, or Pneumonia among Young and Middle-Aged Adults: A Retrospective Observational Cohort Study. |
BACKGROUND: Patients aged >/=65 years are vulnerable to readmissions due to a transient period of generalized risk after hospitalization. However, whether young and middle-aged adults share a similar risk pattern is uncertain. We compared the rate, timing, and readmission diagnoses following hospitalization for heart failure (HF), acute myocardial infarction (AMI), and pneumonia among patients aged 18-64 years with patients aged >/=65 years. METHODS AND FINDINGS: We used an all-payer administrative dataset from California consisting of all hospitalizations for HF (n = 206,141), AMI (n = 107,256), and pneumonia (n = 199,620) from 2007-2009. The primary outcomes were unplanned 30-day readmission rate, timing of readmission, and readmission diagnoses. Our findings show that the readmission rate among patients aged 18-64 years exceeded the readmission rate in patients aged >/=65 years in the HF cohort (23.4% vs. 22.0%, p<0.001), but was lower in the AMI (11.2% vs. 17.5%, p<0.001) and pneumonia (14.4% vs. 17.3%, p<0.001) cohorts. When adjusted for sex, race, comorbidities, and payer status, the 30-day readmission risk in patients aged 18-64 years was similar to patients >/=65 years in the HF (HR 0.99; 95%CI 0.97-1.02) and pneumonia (HR 0.97; 95%CI 0.94-1.01) cohorts and was marginally lower in the AMI cohort (HR 0.92; 95%CI 0.87-0.96). For all cohorts, the timing of readmission was similar; readmission risks were highest between days 2 and 5 and declined thereafter across all age groups. Diagnoses other than the index admission diagnosis accounted for a substantial proportion of readmissions among age groups <65 years; a non-cardiac diagnosis represented 39-44% of readmissions in the HF cohort and 37-45% of readmissions in the AMI cohort, while a non-pulmonary diagnosis represented 61-64% of patients in the pneumonia cohort. CONCLUSION: When adjusted for differences in patient characteristics, young and middle-aged adults have 30-day readmission rates that are similar to elderly patients for HF, AMI, and pneumonia. A generalized risk after hospitalization is present regardless of age. Please see later in the article for the Editors' Summary. |
Development and Validation of a Harmonized TaqMan-Based Triplex Real-Time RT-PCR Protocol for the Quantitative Detection of Normalized Gene Expression Profiles of Seven Porcine Cytokines. |
Dysregulation of cytokine responses plays a major role in the pathogenesis of severe and life-threatening infectious diseases like septicemia or viral hemorrhagic fevers. In pigs, diseases like African and classical swine fever are known to show exaggerated cytokine releases. To study these responses and their impact on disease severity and outcome in detail, reliable, highly specific and sensitive methods are needed. For cytokine research on the molecular level, real-time RT-PCRs have been proven to be suitable. Yet, the currently available and most commonly used SYBR Green I assays or heterogeneous gel-based RT-PCRs for swine show a significant lack of specificity and sensitivity. The latter is however absolutely essential for an accurate quantification of rare cytokine transcripts as well as for detection of small changes in gene expressions. For this reason, a harmonized TaqMan-based triplex real-time RT-PCR protocol for the quantitative detection of normalized gene expression profiles of seven porcine cytokines was designed and validated within the presented study. Cytokines were chosen to represent different immunological pathways and targets known to be involved in the pathogenesis of the above mentioned porcine diseases, namely interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, tumor necrosis factor (TNF)-alpha and interferon (IFN)-alpha. Beta-Actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as reference genes for normalization. For absolute quantification a synthetic standard plasmid was constructed comprising all target cytokines and reference genes within a single molecule allowing the generation of positive control RNA. The standard as well as positive RNAs from samples, and additionally more than 400 clinical samples, which were collected from animal trials, were included in the validation process to assess analytical sensitivity and applicability under routine conditions. The resulting assay allows the reliable assessment of gene expression profiles and provides a broad applicability to any kind of immunological research in swine. |
The individual N- and C-lobes of calmodulin tether to the Cav1.2 channel and rescue the channel activity from run-down in ventricular myocytes of guinea-pig heart. |
The present study examined the binding of the individual N- and C-lobes of calmodulin (CaM) to Cav1.2 at different Ca2+ concentration ([Ca2+]) from approximately free to 2 mM, and found that they may bind to Cav1.2 Ca2+-dependently. In particular, using the patch-clamp technique, we confirmed that the N- or C-lobes can rescue the basal activity of Cav1.2 from run-down, demonstrating the functional relevance of the individual lobes. The data imply that at resting [Ca2+], CaM may tether to the channel with its single lobe, leading to multiple CaM molecule binding to increase the grade of Ca2+-dependent regulation of Cav1.2. |
L Protease from Foot and Mouth Disease Virus Confers eIF2-Independent Translation for Mrnas Bearing Picornavirus Ires. |
The leader protease (Lpro) from foot-and-mouth disease virus (FMDV) has the ability to cleave eIF4G, leading to a blockade of cellular protein synthesis. In contrast to previous reports, our present findings demonstrate that FMDV Lpro is able to increase translation driven by FMDV IRES. Additionally, inactivation of eIF2 subsequent to phosphorylation induced by arsenite or thapsigargin in BHK cells blocks protein synthesis directed by FMDV IRES, whereas in the presence of Lpro, significant translation is found under these conditions. This phenomenon was also observed in cell-free systems after induction of eIF2 phosphorylation by addition of poly(I:C). |
Ambrisentan improves the outcome of rats with liver transplantation partially through reducing nephrotoxicity. |
OBJECTIVE: Tacrolimus is a potent immunosuppressive agent mainly used for allogeneic solid organ transplantation. Although usage of tacrolimus led to a significant increase in short-term allograft survival, the long-term morbidity remains high. Endothelin-1 (ET-1) is reported to be associated with increased vascular resistance, CNI-induced nephrotoxicity and chronic rejection. MATERIALS AND METHODS: In the present study, we first detected the serum and renal ET-1 level of rats treated by tacrolimus and found strong positive correlations were existed between the ET-1 level and the tacrolimus dosage and treated time. Furthermore, we studied the protective effect of ambrisentan in liver transplantation rats when co-administrated with tacrolimus. Healthy inbred male Wistar and Sprague-Dawley (SD) rats were used in this study. The post-operative general condition, transplantation survival time, hepatic aminotransferase, serum IFN-gamma and level kidney injury biochemical index were recorded and compared to evaluate the immune response and outcomes in the recipient rats after liver transplantation. RESULTS: Our results indicate that ambrisentan prevents the changes of ET-1 content in rats of non-operative treatment group and reduced the nephrotoxicity in the rats with liver transplantation. Rats from ambrisentan co-administration group exhibited good postoperative condition and prolonged survival. CONCLUSIONS: Ambrisentan reverted some effects induced by tacrolimus in the kidney and indicated a positive potential for therapeutic benefit. |
Interventional treatment strategies of thoracic aortic pathologies. |
Chronic and acute diseases of the thoracic aorta, including aortic dissection and aortic aneurysm are attracting increasing attention both in the light of an ageing Western and Oriental population and with the proliferation of modern diagnostic imaging modalities. While classical surgical strategies still dominate the care for acute and chronic pathology of the ascending aorta and the proximal arch region, new endovascular concepts are emerging and are likely to evolve as primary treatment strategies for descending aortic pathology in suitable patients constituting the majority of cases. Additionally, aortic arch pathologies are becoming the target of hybrid approaches combining surgical head-vessel debranching and interventional stent-graft implantation in the attempt to improve outcome by avoiding the high risk of open arch repair or complete replacement. Nonetheless, due to the complexity of the underlying vascular disease, every patient should be discussed in a team consisting of cardiologists, cardiac surgeons, anaesthesiologists and radiologists and an individualized therapeutic strategy should be carried out in a center with experience in both endovascular and surgical procedures. |
Correlation between the embolism area and pulmonary arterial systolic pressure as an indicator of pulmonary arterial hypertension in patients with acute pulmonary thromboembolism. |
OBJECTIVE: We tested whether correlation between embolism area and pulmonary arterial pressure may serve as an indicator of pulmonary arterial hypertension in patients with acute pulmonary thromboembolism. PATIENTS AND METHODS: In total, 204 patients who underwent spiral computed tomography (CT) pulmonary angiography and ultrasonic cardiogram were enrolled. The patients were stratified according to their pulmonary arterial systolic pressure into those with normal values, or those with mild, moderate, or severe pulmonary arterial hypertension. The embolism area was quantified using Qanadli score. RESULTS: We found that embolism area correlated positively with pulmonary arterial systolic pressure (r = 0.514). Specifically, embolism areas of four study groups were, respectively, 17.72 +/- 13.61%, 18.25 +/- 13.78%, 33.39 +/- 10.99%, 40.46 +/- 15.75%, obviously increasing along with progression of the disease. Pulmonary arterial systolic pressure was above 41 mm Hg when the embolism area was >33%, and above 70 mm Hg with the embolism area of > 40%. CONCLUSIONS: Assessment of the embolism area is useful in the clinical evaluation of pulmonary arterial systolic pressure, but appears to be more applicable to moderate and severe pulmonary arterial hypertension. |
Usnic acid inhibits ER stress activation through AMPK signaling pathway in rat cardiomyocytes. |
OBJECTIVE: In the present study, we investigated the effects of usnic acid (UA) on the endoplasmic reticulum (ER) stress processes in rat cardiomyocytes. MATERIALS AND METHODS: Gene expression of pro-inflammatory cytokines and activation of ER stress signaling were analyzed. Besides, levels of phosphorylated AMPK were measured to evaluate the mechanisms of UA. Finally, small interfering RNA (siRNA) oligos targeting AMPK subunits were used to determine the roles of AMPK in rat cardiomyocytes treated with UA. RESULTS: We found that UA treatment significantly reduced ER stress activation and expression of pro-inflammatory cytokines, via an AMPK signaling-dependent manner. CONCLUSIONS: UA might be useful to reduce the occurrence of adverse cardiovascular events. |
Vacuum Assisted Closure (VAC) therapyTM as a swiss knife multi-tool for enteric fistula closure: tips and tricks: a pilot study. |
OBJECTIVE: Enterocutaneous fistulas (ECFs) are an uncommon surgical problem, but they are characterized by a difficult management. Vacuum-assisted closure (VAC) therapy is a method utilized for chronic and traumatic wound healing. At first, VAC therapy had been contraindicated in the treatment of intestinal fistulas, but as time went by, VAC therapy revealed itself to be a "Swiss knife multi-tool". This paper presents some clinical cases of enterocutaneous (ECF) and enteroatmospheric fistulas (EAF) treated with VAC therapy. MATERIALS AND METHODS: The history of 8 patients treated for complex fistulas was revised. Four of them presented with enterocutaneous and four with enteroatmospheric fistulas. All were treated with VAC therapy with variations elaborated to help in accelerated closure of intestinal wall lesions. RESULTS: Four out of four ECFs closed spontaneously. In the EAF group, in three cases the fistula turned slowly into an entero-cutaneous fistula, and in one out of four it closed spontaneously. The mean length of VAC therapy was 35.5 days and that of spontaneous closure was 36.4 days. CONCLUSIONS: The results of our study encourage the use of VAC therapy for the treatment of enterocutaneous fistulas. VAC therapy use has a double therapeutic value: (1) it promotes the healing of the skin and allows also the management of EAFs; (2) in selected cases, those in which it is possible to create a deep fistula tract ("well") it is possible to assist to a complete healing with closure of the ECFs. |
Qualitative motor assessment allows to predict the degree of motor disturbances. |
OBJECTIVE: Early diagnosis is necessary in order to determine neurological integrity and the potential risk of improper development, and also to undertake possible early intervention. The quantitative assessment consists of observation of motor development, and provides information about whether a child performs an activity (movement) expected at a given life period. The qualitative assessment of motor performance verifies whether a specific activity is performed properly. The papers aims to demonstrate the motor performance assessment sheet for infants at the age of 6 months and assessment of qualitative elements of 3rd month at the age of 6 months. SUBJECTS AND METHODS: 173 infants (76f/97m) were assessed by a neurologist and a physiotherapist at the age of 6 and 9 months. The neurologist set the final diagnosis at the age of 16 months. Additionally, the physiotherapist assessed qualitative elements typical of the 6th month and of the 3rd month. Risk factors possibly affecting motor performance were considered. RESULTS: The assessment performed by the neurologist and the physiotherapist demonstrated high conformity. Infants with varying degrees of developmental delay and with cerebral palsy at the age of 6 months have still not achieved all of the qualitative characteristics typical of the 3rd month of life, nor proper performance for the 6th month. The low Apgar score and the presence of intraventricular haemorrhage affected the motor development at the age of 6 months. CONCLUSIONS: The author's motor development assessment sheet applied at the age of 6 months proves to be a sensitive research tool and demonstrates good predictive value. |
Bickerstaff's brainstem encephalitis (BBE) in childhood: rapid resolution after intravenous immunoglobulins treatment. |
Three young patients with Bickerstaff's brainstem encephalitis (BBE) are reported. Some weeks following an upper tract infection, the children after a short period of recovery, showed acute onset of symmetric weakness of the lower limbs with difficulty in standing by and walking. The distal muscle weakness had a rapid progression with involvement of the cranial nerve, and then with severe impairment of the consciousness till to coma in one of the three children. BBE is a rare and often underdiagnosed affection in childhood. Common neuro-immune pathogenesis, overlap of clinical signs and strict correlation among BBE with Fisher syndrome and Guillain-Barre syndrome lead to think that these affections represent an unique spectrum with different central and peripheral involvement. In these children, treatment with intravenous immunoglobulins resulted in a progressive and rapid resolution of the clinical features. |
Effects of different antihypertensive drugs on blood pressure variability in patients with ischemic stroke. |
OBJECTIVE: Blood pressure variation is one of the factors that affects the risk of stroke recurrence and prognosis. This study investigates the effects of calcium channel blockers and beta-blockers on blood pressure variability in severe ischemic stroke patients. PATIENTS AND METHODS: The clinical data of 24 patients with ischemic stroke in our intensive care unit were analyzed, and received amlodipine or metoprolol for more than 14 days with 24-hour ambulatory blood pressure monitoring. All patients aged 61-90 years, with GCS score </= 8 or associated with other organ dysfunction. RESULTS: Among these 24 ischemic stroke patients, 12 received amlodipine and 12 received metoprolol. The observation period was divided into two phases: 1-6 days and 7-14 days. The decrease in blood pressure was faster in the metoprolol group than in the amlodipine group, while the average standard deviation was significantly greater and the smoothness index was less. CONCLUSIONS: Metoprolol has faster onset than amlodipine and less blood pressure variability than metoprolol. |
Prevalence of sleep disturbance in chronic pain. |
OBJECTIVE: Sleep is a vital function for human beings, which can be affected by several factors. Chronic pain is one of these factors where it is the most frequent cause for seeking medical care in combination with insomnia. The aim of this study is to examine the prevalence and relationship between sleep disturbance and chronic pain. PATIENTS AND METHODS: After approval, a total of 85 Family Medicine Units from over 170 in Tokat were randomly selected using a 50% sampling. A sample of 2635 subjects, over the age of 19 years, who were registered with the selected Family Medicine Units, were assessed due to gender, age group, and the urban/rural population size of Tokat using the stratified sampling method. The sample size distribution was calculated as 1515 urban subjects, 1120 rural subjects; 1345 female subjects, 1290 male subjects; 1123 subjects between 20-39 years of age, 1103 subjects between the ages of 40-64, and 409 subjects over 64 years of age. After sampling, subjects were invited to participate in the study via an invitation letter, and agreeing individuals were taken to the Family Medicine Unit for face-to-face meetings. Written, informed consent was obtained, along with demographic data. The presence of chronic pain was recorded. According to the presence of chronic pain, all subjects were separated into two groups as Group Chronic Pain and Group Non-Chronic Pain. The visual analog scale for pain intensity, and Pittsburgh Sleep Quality Index for sleep quality, were performed with all subjects. A multiple linear regression model was used to assess the predictors of sleep quality. Analyses were conducted using the Statistical Package for Social Sciences program (SPSS Inc., Chicago, IL, USA), version 20.0. The statistical significance for all analyses was set at p < 0.05. RESULTS: The mean global Pittsburgh Sleep Quality Index score of Group Chronic Pain (5.30 +/- 4.29) was significantly higher than in Group Non-Chronic Pain (3.22 +/- 3.30; p < 0.01). The mean Pittsburgh Sleep Quality Index scores of females (5.69 +/- 4.40) were significantly higher than males (4.54 +/- 3.96) in Group Chronic Pain (p = 0.000045). A total of 40.7% of patients in Group Chronic Pain, and 21.9% in Group Non-Chronic Pain demonstrated poorer sleep quality according to the Pittsburgh Sleep Quality Index scores, with a cut-off level > 5. A moderate positive correlation was found between the global Pittsburgh Sleep Quality Index and Visual Analog Scale scores (r = 0.310, p < 0.01). A multiple linear regression analysis showed that age, gender, income, Visual Analog Scale, and presence of depression were the significant predictors for Pittsburgh Sleep Quality Index score. CONCLUSIONS: The current study revealed that chronic pain and pain intensity are important predictors of sleep quality. |
Agents for refractory/relapsed acute lymphocytic leukemia in adults. |
Although treatment results for adult acute lymphoblastic leukemia (ALL) have improved considerably in the past decades, treating adult patients with relapsed/refractory acute lymphocytic leukemia (ALL) is still difficult. Adults with refractory/relapsed acute lymphocytic leukemia (ALL) processed to death rapidly associated with chemotherapy resistance, high mortality by reinduction, etc. Only 20% to 30% of those patients acquired complete remission (CR). Those patients are always of short duration unless an allogeneic stem cell transplant is feasible. Median survival is only ranging from 2 to 12 months. Therapeutic strategy on relapsed/refractory acute lymphocytic leukemia (ALL) is always a major therapeutic challenge bothering hematological researchers. Novel agents and unique therapeutic strategies have been developed in recent years. This review focuses on major clinical advances in the agents for refractory/relapsed ALL. |
Efficacy of intratumoral chemotherapy using arsenic trioxide (As2O3) sustained release tablets for the treatment of neurogliocytoma in nude mice. |
OBJECTIVE: This study aimed to investigate the inhibiting effect of arsenic trioxide (As2O3) on neurogliocytoma in nude mice and the mechanism responsible for this effect. MATERIALS AND METHODS: Neurogliocytoma implantation models were constructed in nude mice, which were assigned to three groups: the control group, the sustained release tablet-polylactic acid-glycolic acid polymer (50:50) group (PLGA group) and the As2O3-polylactic acid-glycolic acid polymer (50:50) (As2O3-PLGA group). One tablet of As2O3-PLGA was implanted in the tumor of the As2O3-PLGA group. Intratumoral implantation was also performed in the other groups using a different type of tablet. The sustained releasing As2O3 had an inhibiting effect on the tumors. The TUNEL assay was used to determine the apoptosis rates in the implanted tumors. Immunohistochemical staining and Western blotting was carried out to determine the expression levels of caspase-3 and Bcl-2. RESULTS: No inhibitory effect was observed on the tumor in the PLGA group, and there was no significant difference between this group and the control group. Subcutaneous tumor growth in nude mice was significantly inhibited in the As2O3-PLGA group relative to that in the control group, and the difference was statistically significant (p < 0.01). The tumor inhibition rate was 60.8%. The percentage of apoptotic tumor cells in the As2O3-PLGA group was 30.8%, which was significantly higher than that in the control group (3.92%) and that in the PLGA group (4.08%). The expression of Bcl-2 in the implanted tumor tissue was significantly reduced, but the expression of caspase-3 increased significantly. CONCLUSIONS: As2O3 has a potent inhibiting effect on the growth of neurogliocytoma in vivo and can induce the apoptosis of tumor cells. The molecular mechanism of this effect may be related to the downregulation of Bcl-2 expression and the upregulation of caspase-3 expression. |
Saikosaponin-d inhibits proliferation of human undifferentiated thyroid carcinoma cells through induction of apoptosis and cell cycle arrest. |
OBJECTIVE: Saikosaponin-d is a triterpene saponin derived from Bupleurum falcatum. L and has been reported to exhibit a variety of pharmacological activities such as anti-bacterial, anti-virus and anti-cancer. The aim of the present study was to explore the effect of saikosaponin-d on the proliferation and apoptosis of human undifferentiated thyroid carcinoma. MATERIALS AND METHODS: Three human anaplastic thyroid cancers cell lines were cultured in the presence of saikosaponin-d and their proliferation was measured by MTT assay. Cell apoptosis and cell cycle distribution were analyzed with flow cytometry. Western blot was performed to determine the proteins expression. The in vivo effect of saikosaponin-d was measured with an animal model. RESULTS: In vitro, MTT assay showed that saikosaponin-d treatment inhibited cell proliferation in three human anaplastic thyroid cancers cell lines ARO, 8305C and SW1736. In addition, saikosaponin-d promoted cell apoptosis and induced G1-phase cell cycle arrest as shown by flow cytometric analysis. On the molecular level, our results showed that saikosaponin-d treatment increased the expression of p53 and bax, and decreased the expression of Bcl-2. In addition, saikosaponin-d administration led to a significant up-regulation of p21 and down-regulation of CDK2 and cyclin D1. Xenografts tumorigenesis model demonstrated that saikosaponin-d markedly reduced the weight and volume of thyroid tumors in vivo. CONCLUSIONS: The present study suggested that saikosaponin-d might be a new potent chemopreventive drug candidate for human undifferentiated thyroid carcinoma through induction of apoptosis and cell cycle arrest. |
Wortmannin enhances cisplatin-induced apoptosis in human ovarian cancer cells in vitro. |
OBJECTIVE: The PI3K/Akt signaling pathway is constitutively activated in some ovarian cancers; when activated, it promotes invasion and inhibits chemotherapy-mediated apoptosis in cancer cells. The fungal metabolite wortmannin is the currently known inhibitors that show fairly high specificity for PI3K.We examined whether PI3K/Akt activity correlates with invasion and apoptotic resistance to chemotherapy in cultured human ovarian cancer cells, and whether inhibition of PI3K/Akt by wortmannin inhibits invasion and enhances cisplatin-induced apoptosis in cultured human ovarian cancer cells. MATERIALS AND METHODS: The cisplatin-sensitive A2780 ovarian adenocarcinoma cell line and its daughter line, A2780cis was evaluated for basal Akt activity. Chemotherapy-induced cell death was evaluated following down-regulation of Akt activity by wortmannin treatment or upregulation of Akt activity by myr-Akt treatment. Invasion and migration were assessed using Boyden chamber assays RESULTS: Inhibiting or activation of PI3K/Akt signaling pathway by wortmannin had little effect on the basal level of apoptosis in ovarian cancer cells, but increased the apoptotic effect of chemotherapy in A2780cis cells, decreased the apoptotic effect of chemotherapy in cisplatin-sensitive A2780 cells. Cisplatin resistant cells display increased potential for migration and invasion. CONCLUSIONS: The antiapoptotic effect of AKT activation in ovarian cancer cells confer invasive ability and resistance to apoptosis. Wortmannin is as adjuncts to conventional chemotherapy in the treatment of ovarian cancer. |
The role of gut microbiota in the pathogenesis and management of allergic diseases. |
This article has been withdrawn. |
Autologous pericardium annuloplasty: a "physiological" mitral valve repair. |
AIM: Autologous pericardium annuloplasty (APA) is an alternative to prosthetic ring implantation for mitral valve (MV) repair, avoiding the use of foreign material and preserving the mitral annulus' physiological motion. However, data on durability are questionable. Therefore, we analyzed long-term outcomes of treating degenerative mitral regurgitation (MR) with APA. METHODS: Four hundred ninety patients (mean age, 54.3+/-11.3 years, [15-77 years]; N.=360 men [74.1%]) who had undergone APA and neochordae implantation between July 1988 and December 2006 were retrospectively studied. RESULTS: MR was purely degenerative in 434 (89.3%) patients; endocarditis was present in 44 (9.1%) patients; an anterior, posterior, or bileaflet prolapse was present in 32 (6.6%), 241 (49.6%), and 213 (43.8%) patients, respectively. Clinical follow-up was 100% complete at a median of 6.5 years (5th percentile, 0.9; 95th percentile, 14.9) with an echocardiographic study in 92% of patients. In-hospital mortality was 1% (5 deaths); overall and late cardiac mortality were 7.6% and 3.9% (37 and 19 deaths), respectively. Kaplan-Meier curves for overall survival, late cardiac survival, and freedom from reoperation at 15 years (20 cases) were 86% (95%CI 80-91), 93% (95%CI 88-96), and 93% (95%CI 88-96), respectively. At 15 years, freedom from recurrent MR (28 patients) and endocarditis (6 events) were 86% (95%CI 76-91) and 97% (95%CI 92-99). Dehiscence, significant calcification of APA, and hemolysis never occurred. At reoperations, annular pericardium appeared covered by a smooth layer of tissue. CONCLUSION: APA is feasible, safe, and cost-effective, providing long-term durability, high survival, and a low rate of valve-related complications. |
BRAF alterations in brain tumours: molecular pathology and therapeutic opportunities. |
PURPOSE OF REVIEW: To summarize the current knowledge on v-RAF murine sarcoma viral oncogene homologue B1 (BRAF) aberrations in tumours of the central nervous system. RECENT FINDINGS: BRAF alterations are found in variable frequencies across a wide spectrum of diverse central nervous system neoplasms. BRAF V600 point mutations (most commonly of the V600E type) are most common in pleomorphic xanthoastrocytoma (approximately 60% of cases), gangliogliomas (50%), dysembryoplastic neuroepithelial tumours (30%), Langerhans cell histiocytosis (50%), melanoma brain metastases (50%) and papillary craniopharyngiomas (96%) and are also detectable in a fraction of glioblastomas (overall mutation rate of 2-12%, with a higher rate of approximately 50% in epithelioid glioblastomas). BRAF fusions (most commonly KIAA1549: BRAF) are typical for pilocytic astrocytomas and are almost absent from other tumour types. Clinical trials have established tyrosine-kinase inhibitors of BRAF as feasible treatment option in selected patients with mutation-bearing brain metastases of melanoma. Preclinical studies, some case reports and small patient series have documented tumour responses of primary brain tumours with BRAF aberrations to BRAF inhibition. SUMMARY: Molecular testing for BRAF alterations in brain tumours may be of clinical relevance for differential diagnostic considerations in some situations or to guide selection of patients for targeted therapy with specific inhibitors. Prospective clinical trials evaluating the efficacy of BRAF inhibitors in central nervous system tumours are strongly supported by the available evidence. |
Prognostic Scoring Models for Patients Undergoing Sorafenib Treatment for Advanced Stage Hepatocellular Carcinoma in Real-Life Practice. |
BACKGROUND:: The purpose of this study was to build prognostic models capable of estimating the outcomes of individual sorafenib-treated advanced stage hepatocellular carcinoma (HCC) patients based on specific patient and tumor factors. METHODS:: A parametric model for time-to-event data was used to construct scoring systems based on the intent-to-treat data set from 480 sorafenib-treated patients with advanced stage HCC: 356 for derivation and 124 for validation. Clinical parameters included in the models were based on importance variable scores generated by a random forest approach and bootstrap resampling. The model's accuracy was internally and externally assessed using the time-dependent C-index of discrimination and a Hosmer-Lemeshow type test for calibration. RESULTS:: The models generated for time-to-progression and overall survival based on Child-Pugh score, serum alpha-fetoprotein, tumor morphology, and vascular invasion and/or extrahepatic involvement had good calibration and discrimination abilities, with C-indexes of 0.669 (3 mo progression) and 0.809 (6 mo survival), respectively. External validation results also showed that these models performed well in terms of goodness-of-fit and discrimination (C-index: 0.746 for 3 mo progression and 0.875 for 6 mo survival). Receiver operating characteristic curve analysis in the validation patients indicated that these models have better predictive power than Child-Pugh scores (C-index: 0.686 for 3 mo progression and 0.777 for 6 mo survival). CONCLUSIONS:: The prognostic tools developed to quantify the potential outcomes for progression and survival expected from sorafenib treatment can serve as useful clinical aids in personalized decision making regarding treatment in advanced stage HCC patients. |
Hepatocellular Carcinoma Management in Nonalcoholic Fatty Liver Disease Patients: Applicability of the BCLC Staging System. |
BACKGROUND/AIMS:: Nonalcoholic fatty liver disease (NAFLD) has emerged as an important cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma (HCC). The Barcelona Clinic Liver Cancer (BCLC) system is the preferred staging system to evaluate patients with HCC and links prognosis assessment with treatment recommendation. The aim of this retrospective study was to evaluate whether the BCLC staging system and its treatment algorithm are suitable for patients with HCC arising from NAFLD. METHODS:: Forty-two patients with HCC related to either to NAFLD or cryptogenic cirrhosis were retrieved retrospectively from 2 centers in Brazil. Patients were classified according to BCLC staging system. If the proposed HCC therapy could not be applied, the case was considered to represent deviations from the recommended BCLC guideline. Causes of treatment deviations were investigated. RESULTS:: There were 4 patients without evidence of cirrhosis according to liver biopsy and/or clinical evaluation. One (2%), 21 (50%), 10 (24%), 5 (12%), and 5 patients (12%) were classified initially to the very early (0), early (A), intermediate (B), advanced (C), and terminal (D) BCLC stages, respectively. Thirty-five patients (83%) were treated according to BCLC recommendations. There were 3 cases (of 5) of protocol deviation in BCLC C patients. The 1- and 2-year overall survival rates were 81% and 66%, respectively. CONCLUSIONS:: The BCLC system is applied in most cases of NAFLD-related HCC cases. Deviation of BCLC is found more frequently in BCLC C stage patients.This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivitives 3.0 License, where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially. http://creativecommons.org/licenses/by-nc-nd/3.0. |
Late Toxicity and Outcomes in High-risk Prostate Cancer Patients Treated With Hypofractionated IMRT and Long-term Androgen Suppression Treatment. |
OBJECTIVE:: To assess late toxicity and outcomes in high-risk prostate cancer patients treated with hypofractionated radiation treatment with androgen suppression therapy. METHODS:: Sixty high-risk prostate cancer patients were enrolled. IMRT prescription was 68 Gy/25 fractions (2.7 Gy/fraction) to the prostate and proximal seminal vesicles (SV). The pelvic lymph nodes (PLN) and distal SV concurrently received 45 Gy/25 fractions (1.8 Gy/fraction). The patients were treated with helical TomoTherapy-based IMRT and underwent daily megavoltage CT image-guided verification before each treatment. RTOG Toxicity scores were recorded for a 5-year period. RESULTS:: Sixty patients completed RT with median follow-up of 63 months (range, 7 to 80 mo).At 5 years follow-up timepoint: Grade (G)2 and G3 late genitourinary toxicity was experienced in 7 (17.0%) and 1 (2.44%), respectively; gastrointestinal G2 as highest toxicity recorded in only 1 (2.44%) patient. There was no G3 gastrointestinal toxicity recorded at this timepoint.With 63-month median follow-up (mean of 65.41+/-1.72 mo), the 5-year overall survival was 86.67%; 5 years freedom from biochemical failure was 91.67% and freedom from clinical failure was 96.67%. CONCLUSIONS:: Dose escalation and hypofractionated radiation treatment with IMRT treating the prostate and proximal SV concurrently with the pelvic lymph nodes and distal SV and long-term androgen suppression therapy is well tolerated with respect to acute and late toxicity with 5-year actuarial overall survival 86.67%, freedom from biochemical failure 91.38%, and freedom from clinical failure 96.67%. Longer follow-up will provide more information on 10-year survival outcomes. |
Galangin suppresses HepG2 cell proliferation by activating the TGF-beta receptor/Smad pathway. |
Galangin can suppress hepatocellular carcinoma (HCC) cell proliferation. In this study, we demonstrated that galangin induced autophagy by activating the transforming growth factor (TGF)-beta receptor/Smad pathway and increased TGF-beta receptor I (RI), TGF-betaRII, Smad1, Smad2, Smad3 and Smad4 levels but decreased Smad6 and Smad7 levels. Autophagy induced by galangin appears to depend on the TGF-beta receptor/Smad signalling pathway because the down-regulation of Smad4 by siRNA or inhibition of TGF-beta receptor activation by LY2109761 blocked galangin-induced autophagy. The down-regulation of Beclin1, autophagy-related gene (ATG) 16L, ATG12 and ATG3 restored HepG2 cell proliferation and prevented galangin-induced apoptosis. Our findings indicate a novel mechanism for galangin-induced autophagy via activation of the TGF-beta receptor/Smad pathway. The induction of autophagy thus reflects the anti-proliferation effect of galangin on HCC cells. |
Comparison of Propofol, Etomidate, and Thiopental in Anesthesia for Electroconvulsive Therapy: A Randomized, Double-blind Clinical Trial. |
OBJECTIVES: This study aimed to compare the effects of propofol, thiopental, and etomidate, which are routinely used in anesthesia for electroconvulsive therapy (ECT), on the cardiovascular system, seizure variables, recovery, cognitive functions, and response to treatment. METHODS: Male patients hospitalized at the Seventh Psychiatry Clinics of the Bakirkoy Teaching Hospital for Psychiatry, Neurology, and Neurosurgery who were treated with ECT were investigated prospectively. The effects on cardiovascular system parameters (heart rate, blood pressure, and blood oxygenation), seizure variables (duration and intensity of seizure), and recovery variables were recorded at every session, on prespecified time points, and the findings of the first session were used in this evaluation. In addition, clinical responses to treatment were evaluated with tests of cognitive functions before and after a course of ECT. Adverse effects were recorded. RESULTS: The sociodemographic characteristics of the 3 treatment groups were similar. There were no significant differences among the groups in terms of effects on cardiovascular system variables, seizure variables, and cognitive functions. The clinical response to ECT was good in all groups, without any significant differences. CONCLUSIONS: Propofol, etomidate, and thiopental are associated with similar safety and efficacy profiles. |
Direct effects of recurrent hypoglycaemia on adrenal catecholamine release. |
In Type 1 and advanced Type 2 diabetes mellitus, elevation of plasma epinephrine plays a key role in normalizing plasma glucose during hypoglycaemia. However, recurrent hypoglycaemia blunts this elevation of plasma epinephrine. To determine whether recurrent hypoglycaemia affects peripheral components of the sympatho-adrenal system responsible for epinephrine release, male rats were administered subcutaneous insulin daily for 3 days. These recurrent hypoglycaemic animals showed a smaller elevation of plasma epinephrine than saline-injected controls when subjected to insulin-induced hypoglycaemia. Electrical stimulation of an adrenal branch of the splanchnic nerve in recurrent hypoglycaemic animals elicited less release of epinephrine and norepinephrine than in controls, without a change in adrenal catecholamine content. Responsiveness of isolated, perfused adrenal glands to acetylcholine and other acetylcholine receptor agonists was also unchanged. These results indicate that recurrent hypoglycaemia compromised the efficacy with which peripheral neuronal activity stimulates adrenal catecholamine release and demonstrate that peripheral components of the sympatho-adrenal system were directly affected by recurrent hypoglycaemia. |
Utility of galectin-3 as a prognostic biomarker in heart failure: where do we stand? |
BACKGROUND: Heart failure (HF) continues to be an illness of daunting proportions with a four- year mortality touching 50%. Biomarkers for prognosticating patients with heart failure have generated immense interest. Several studies have been conducted on a novel biomarker, galectin-3 to assess its prognostic effect in heart failure populations. However, the studies have generated conflicting results. Hence a systematic review was done to assess the utility of galectin-3 as a prognostic biomarker in HF. DESIGN: This study was a systematic review. METHODS: A literature search was done using terms 'galectin-3 and heart' and 'galectin-3 and heart failure' in MEDLINE, Science Direct, Scopus, Springer Link, Cochrane Library and Google Scholar for original articles using a predefined inclusion/exclusion criteria. RESULTS: Altogether 27 original articles were selected for the systematic review. Multivariate analysis showed galectin-3 to be ineffective in predicting all-cause mortality and cardiovascular mortality especially under the influence of factors such as estimated glomerular filtration rate (eGFR), left ventricular ejection fraction (LVEF) and N-terminal pro-B-type natriuretic peptide (NTproBNP). Galectin-3 was not found to be superior to NTproBNP, sST2, GDF-15 or C-reactive protein (CRP) as a predictor of mortality. However the combination of natriuretic peptides and galectin-3 has been observed to be superior in predicting mortality compared to either of the biomarkers alone. The role of galectin-3 in remodelling has not been conclusively proven as seen in earlier pre-clinical studies. CONCLUSION: The current weight of evidence does not suggest galectin-3 to be a predictor of mortality. However, assessment of galectin-3 in a multi-biomarker panel may have a distinct advantage in prognosticating patients with heart failure. |
The Yale-Brown Obsessive Compulsive Scale: A Reliability Generalization Meta-Analysis. |
The Yale-Brown Obsessive Compulsive Scale (Y-BOCS) is the most frequently applied test to assess obsessive compulsive symptoms. We conducted a reliability generalization meta-analysis on the Y-BOCS to estimate the average reliability, examine the variability among the reliability estimates, search for moderators, and propose a predictive model that researchers and clinicians can use to estimate the expected reliability of the Y-BOCS. We included studies where the Y-BOCS was applied to a sample of adults and reliability estimate was reported. Out of the 11,490 references located, 144 studies met the selection criteria. For the total scale, the mean reliability was 0.866 for coefficients alpha, 0.848 for test-retest correlations, and 0.922 for intraclass correlations. The moderator analyses led to a predictive model where the standard deviation of the total test and the target population (clinical vs. nonclinical) explained 38.6% of the total variability among coefficients alpha. Finally, clinical implications of the results are discussed. |
The Effectiveness of the Personality Assessment Inventory With Feigned PTSD: An Initial Investigation of Resnick's Model of Malingering. |
Malingered posttraumatic stress disorder (PTSD) poses a formidable clinical challenge because of the apparent ease in feigning PTSD. As an additional confound, some patients with genuine PTSD produce elevated profiles on feigning indicators that are difficult to distinguish from feigned PTSD. The current study utilized 109 inpatients from a trauma unit to examine whether the Personality Assessment Inventory and the Detailed Assessment of Posttraumatic Stress can effectively differentiate between genuine and feigned PTSD. As a primary focus, Resnick's model of malingered PTSD was evaluated with its three subtypes: pure malingering, partial malingering, and false imputation. They were tested on their ability to (a) effectively simulate PTSD and (b) avoid being classified as feigning. The partial malingering group proved to be the best feigning group in achieving these two goals. Overall, the Personality Assessment Inventory Malingering Index and Negative Distortion Scale were the most effective at identifying feigning. |
Renal Sulfatides: Sphingoid Base-Dependent Localization and Region-Specific Compensation of CerS2-Dysfunction. |
Mammalian kidneys are rich in sulfatides. Especially papillary sulfatides contribute to renal adaptation to chronic metabolic acidosis. Due to differences in their ceramide anchors, the structural diversity of renal sulfatides is large. However, the underling biological function of this complexity is not understood. As a compound's function and its tissue location are intimately connected, we analyzed individual renal sulfatide distributions of control and ceramide synthase 2-deficient mice by imaging mass spectrometry and by LC-MS2, (in controls for cortex, medulla, and papillae separately). To explain locally different structures, we compared our lipid data with regional mRNA levels of corresponding anabolic enzymes. The combination of IMS and in source decay-LC-MS2 analyses revealed exclusive expression of C20-sphingosine-containing sulfatides within renal papillae, whereas conventional C18-sphingosine-containing compounds were predominant in medulla, and sulfatides with a C18-phytosphingosine were restricted to special cortical structures. Ceramide synthase 2 deletion resulted in bulk loss of sulfatides with C23/C24-acyl chains, but did not lead to decreased urinary pH as previously observed in sulfatide-depleted kidneys. Reasons may be the almost unchanged C22-sulfatid levels and constant total renal sulfatide levels due to compensation with C16- to C20-acyl chain-containing compounds. Intriguingly, CerS2-deficient kidneys were completely depleted of phytosphingosine-containing cortical sulfatides without any compensation. |
Phase II Study of Concurrent Chemoradiotherapy with S-1 in Patients with Stage II (T2N0M0) Squamous Cell Carcinoma of the Pharynx or Larynx. |
OBJECTIVE: The goals of treatment for head and neck cancer are cure and organ-function preservation. For organ preservation, primary treatment via radiotherapy alone is thought to be insufficient for Stage II squamous cell carcinoma of the larynx, oropharynx or hypopharynx. The objective of the present study was to investigate the efficacy and safety of concurrent chemoradiotherapy with S-1 for patients with Stage II squamous cell carcinoma of the pharynx or larynx for primary organ preservation. METHODS: Previously untreated patients with Stage II squamous cell carcinoma of the larynx, oropharynx or hypopharynx received three courses of S-1 (40 or 50 mg twice a day; 2 weeks of administration followed by 1 week of rest every 3 weeks) during conventional radiotherapy (a single daily fraction of 1.8 Gy) to a total dose of 70.2 Gy. The primary endpoint was the local control rate at 3 years. RESULTS: From August 2009 to October 2012, 37 patients were evaluated for the study. The overall response rate was 100%. The 3-year local control rate was 89.0% (95% confidence interval, 78.9-99.2%), and the 3-year overall survival rate was 97.2% (95% confidence interval, 91.8-100%). Mucositis and dermatitis in the radiation field were the most common acute adverse events observed. The rates of Grade 3 mucositis and dermatitis were 27 and 35%, respectively. No patients experienced Grade 4 acute adverse events. The treatment completion rate was 89.2%. CONCLUSION: Concurrent chemoradiotherapy with S-1 was safe and effective in improving local control for Stage II squamous cell carcinoma of the pharynx or larynx. |
Efficacy and safety of triple combination therapy with artesunate-amodiaquine-methylene blue for falciparum malaria in children: a randomised controlled trial in Burkina Faso. |
BACKGROUND: Methylene blue (MB) has been shown to be safe and effective against falciparum malaria in Africa and to have pronounced gametocytocidal properties. METHODS: Three days treatment with artesunate (AS)-amodiaquine (AQ)-MB was compared to AS-AQ in a randomised controlled phase IIb study in 221 children aged 6-59 months with uncomplicated falciparum malaria in Burkina Faso. The primary endpoint was gametocyte prevalence during follow-up determined by microscopy and by real-time quantitative nucleic acid sequence-based amplification (QT-NASBA). RESULTS: Gametocyte prevalence of P. falciparum at baseline was 3.6% (microscopy) and 97% (QT-NASBA). Gametocyte prevalence was significantly lower in the AS-AQ-MB compared to AS-AQ group on day 7 of follow-up (1.2% vs. 8.9% by microscopy, p<0.05; 36.7% vs. 63.3% by QT-NASBA, p<0.001). Haemoglobin values were significantly lower in the AS-AQ-MB group compared to AS-AQ at day 2 and 7 of follow-up. Vomiting of the study medication occurred significantly more frequently in children having been treated with AS-AQ-MB compared to AS-AQ. CONCLUSIONS: The combination of MB with an ACT has been confirmed to be effective against the gametocytes of P. falciparum. MB-based combinations need to be compared to primaquine-based combinations, preferably using MB in an improved pediatric formulation. |
Innate Inflammation Induced by the 8-Oxoguanine DNA Glycosylase-1-KRAS-NF-kappaB Pathway. |
8-Oxoguanine-DNA glycosylase-1 (OGG1) is the primary enzyme for repairing 7,8-dihydro-8-oxoguanine (8-oxoG) via the DNA base excision repair pathway (OGG1-BER). Accumulation of 8-oxoG in the genomic DNA leads to genetic instability and carcinogenesis and is thought to contribute to the worsening of various inflammatory and disease processes. However, the disease mechanism is unknown. In this study, we proposed that the mechanistic link between OGG1-BER and proinflammatory gene expression is OGG1's guanine nucleotide exchange factor activity, acquired after interaction with the 8-oxoG base and consequent activation of the small GTPase RAS. To test this hypothesis, we used BALB/c mice expressing or deficient in OGG1 in their airway epithelium and various molecular biological approaches, including active RAS pulldown, reporter and Comet assays, small interfering RNA-mediated depletion of gene expression, quantitative RT-PCR, and immunoblotting. We report that the OGG1-intiated repair of oxidatively damaged DNA is a prerequisite for GDP-->GTP exchange, KRAS-GTP-driven signaling via MAP kinases and PI3 kinases and mitogen-stress-related kinase-1 for NF-kappaB activation, proinflammatory chemokine/cytokine expression, and inflammatory cell recruitment to the airways. Mice deficient in OGG1-BER showed significantly decreased immune responses, whereas a lack of other Nei-like DNA glycosylases (i.e., NEIL1 and NEIL2) had no significant effect. These data unveil a previously unidentified role of OGG1-driven DNA BER in the generation of endogenous signals for inflammation in the innate signaling pathway. |
Mast Cells Control the Expansion and Differentiation of IL-10-Competent B Cells. |
The discovery of B cell subsets with regulatory properties, dependent on IL-10 production, has expanded our view on the mechanisms that control inflammation. Regulatory B cells acquire the ability to produce IL-10 in a stepwise process: first, they become IL-10 competent, a poised state in which B cells are sensitive to trigger signals but do not actually express the Il-10 gene; then, when exposed to appropriate stimuli, they start producing IL-10. Even if the existence of IL-10-competent B cells is now well established, it is not yet known how different immune cell types cross talk with B cells and affect IL-10-competent B cell differentiation and expansion. Mast cells (MCs) contribute to the differentiation and influence the effector functions of various immune cells, including B lymphocytes. In this study, we explored whether MCs could play a role in the expansion of IL-10-competent B cells and addressed the in vivo relevance of MC deficiency on the generation of these cells. We show that MCs can expand IL-10-competent B cells, but they do not directly induce IL-10 production; moreover, the absence of MCs negatively affects IL-10-competent B cell differentiation. Noteworthy, our findings reveal that the CD40L/CD40 axis plays a significant role in MC-driven expansion of IL-10-competent B cells in vitro and highlight the importance of MC CD40L signaling in the colon. |
Neutrophils Counteract Autophagy-Mediated Anti-Inflammatory Mechanisms in Alveolar Macrophage: Role in Posthemorrhagic Shock Acute Lung Inflammation. |
Acute lung injury (ALI) is a major component of multiple organ dysfunction syndrome after hemorrhagic shock (HS) resulting from major surgery and trauma. The increased susceptibility in HS patients to the development of ALI suggests not yet fully elucidated mechanisms that enhance proinflammatory responses and/or suppress anti-inflammatory responses in the lung. Alveolar macrophages (AMvarphi) are at the center of the pathogenesis of ALI after HS. We have previously reported that HS-activated polymorphonuclear neutrophils (PMNs) interact with macrophages to influence inflammation progress. In this study, we explore a novel function of PMNs regulating AMvarphi anti-inflammatory mechanisms involving autophagy. Using a mouse "two-hit" model of HS/resuscitation followed by intratracheal injection of muramyl dipeptide, we demonstrate that HS initiates high mobility group box 1/TLR4 signaling, which upregulates NOD2 expression in AMvarphi and sensitizes them to subsequent NOD2 ligand muramyl dipeptide to augment lung inflammation. In addition, upregulated NOD2 signaling induces autophagy in AMvarphi, which negatively regulates lung inflammation through feedback suppression of NOD2-RIP2 signaling and inflammasome activation. Importantly, we further demonstrate that HS-activated PMNs that migrate in alveoli counteract the anti-inflammatory effect of autophagy in AMvarphi, possibly through NAD(P)H oxidase-mediated signaling to enhance I-kappaB kinase gamma phosphorylation, NF-kappaB activation, and nucleotide-binding oligomerization domain protein 3 inflammasome activation, and therefore augment post-HS lung inflammation. These findings explore a previously unidentified complexity in the mechanisms of ALI, which involves cell-cell interaction and receptor cross talk. |
Inflammasome Activation by Campylobacter jejuni. |
The Gram-negative pathogen Campylobacter jejuni is the most common cause of bacterial foodborne disease worldwide. The mechanisms that lead to bacterial invasion of eukaryotic cells and massive intestinal inflammation are still unknown. In this study, we report that C. jejuni infection of mouse macrophages induces upregulation of pro-IL-1beta transcript and secretion of IL-1beta without eliciting cell death. Immunoblotting indicated cleavage of caspase-1 and IL-1beta in infected cells. In bone marrow-derived macrophages from different knockout mice, IL-1beta secretion was found to require NLRP3, ASC, and caspase-1/11 but not NLRC4. In contrast to NLRP3 activation by ATP, C. jejuni activation did not require priming of these macrophages. C. jejuni also activated the NLRP3 inflammasome in human macrophages as indicated by the presence of ASC foci and caspase-1-positive cells. Analysis of a vast array of C. jejuni mutants with defects in capsule formation, LPS biosynthesis, chemotaxis, flagella synthesis and flagellin (-like) secretion, type 6 secretion system needle protein, or cytolethal distending toxin revealed a direct correlation between the number of intracellular bacteria and NLRP3 inflammasome activation. The C. jejuni invasion-related activation of the NLRP3 inflammasome without cytotoxicity and even in nonprimed cells extends the known repertoire of bacterial inflammasome activation and likely contributes to C. jejuni-induced intestinal inflammation. |
Variation and Genetic Control of Gene Expression in Primary Immunocytes across Inbred Mouse Strains. |
To determine the breadth and underpinning of changes in immunocyte gene expression due to genetic variation in mice, we performed, as part of the Immunological Genome Project, gene expression profiling for CD4+ T cells and neutrophils purified from 39 inbred strains of the Mouse Phenome Database. Considering both cell types, a large number of transcripts showed significant variation across the inbred strains, with 22% of the transcriptome varying by 2-fold or more. These included 119 loci with apparent complete loss of function, where the corresponding transcript was not expressed in some of the strains, representing a useful resource of "natural knockouts." We identified 1222 cis-expression quantitative trait loci (cis-eQTL) that control some of this variation. Most (60%) cis-eQTLs were shared between T cells and neutrophils, but a significant portion uniquely impacted one of the cell types, suggesting cell type-specific regulatory mechanisms. Using a conditional regression algorithm, we predicted regulatory interactions between transcription factors and potential targets, and we demonstrated that these predictions overlap with regulatory interactions inferred from transcriptional changes during immunocyte differentiation. Finally, comparison of these and parallel data from CD4+ T cells of healthy humans demonstrated intriguing similarities in variability of a gene's expression: the most variable genes tended to be the same in both species, and there was an overlap in genes subject to strong cis-acting genetic variants. We speculate that this "conservation of variation" reflects a differential constraint on intraspecies variation in expression levels of different genes, either through lower pressure for some genes, or by favoring variability for others. |
A Novel Method Using Blinatumomab for Efficient, Clinical-Grade Expansion of Polyclonal T Cells for Adoptive Immunotherapy. |
Current treatment of chronic lymphocytic leukemia (CLL) patients often results in life-threatening immunosuppression. Furthermore, CLL is still an incurable disease due to the persistence of residual leukemic cells. These patients may therefore benefit from immunotherapy approaches aimed at immunoreconstitution and/or the elimination of residual disease following chemotherapy. For these purposes, we designed a simple GMP-compliant protocol for ex vivo expansion of normal T cells from CLL patients' peripheral blood for adoptive therapy, using bispecific Ab blinatumomab (CD3 x CD19), acting both as T cell stimulator and CLL depletion agent, and human rIL-2. Starting from only 10 ml CLL peripheral blood, a mean 515 x 106 CD3+ T cells were expanded in 3 wk. The resulting blinatumomab-expanded T cells (BET) were polyclonal CD4+ and CD8+ and mostly effector and central memory cells. The Th1 subset was slightly prevalent over Th2, whereas Th17 and T regulatory cells were <1%. CMV-specific clones were detected in equivalent proportion before and after expansion. Interestingly, BET cells had normalized expression of the synapse inhibitors CD272 and CD279 compared with starting T cells and were cytotoxic against CD19+ targets in presence of blinatumomab in vitro. In support of their functional capacity, we observed that BET, in combination with blinatumomab, had significant therapeutic activity in a systemic human diffuse large B lymphoma model in NOD-SCID mice. We propose BET as a therapeutic tool for immunoreconstitution of heavily immunosuppressed CLL patients and, in combination with bispecific Ab, as antitumor immunotherapy. |
Progression of Relapsing-Remitting Demyelinating Disease Does Not Require Increased TCR Affinity or Epitope Spread. |
In this study, we investigate the basis of T cell recognition of myelin that governs the progression from acute symptoms into disease remission, relapse, and chronic progression in a secondary progressive model of demyelinating disease. Until now, the frequency and affinity of myelin-reactive CD4 T cells that elicit relapsing-remitting disease have not been quantified. The micropipette adhesion frequency assay was used to obtain a sensitive and physiologically relevant two-dimensional measurement of frequency and TCR affinity for myelin, as the inherent low affinity does not allow the use of specific peptide:MHC-II tetramers for this purpose. We found the highest affinity and frequency of polyclonal myelin oligodendrocyte glycoprotein-reactive cells infiltrate the CNS during acute disease, whereas affinities during remission, relapse, and chronic disease are not significantly different from each other. Frequency analysis revealed that the vast majority of CNS-infiltrating CD4 T cells are myelin oligodendrocyte glycoprotein reactive at all time points, demonstrating epitope spread is not a predominant factor for disease progression. Furthermore, time points at which mice were symptomatic were characterized by an infiltration of Th17 cells in the CNS, whereas symptom remission showed an enrichment of cells producing IFN-gamma. Also, the ratio of regulatory T cells to Foxp3- CD4 T cells was significantly higher in the CNS at remission than during acute disease. The results of this study indicate that a high frequency of T cells specific for a single myelin Ag, rather than increased TCR affinity or epitope spread, governs the transition from acute symptoms through remission, relapse, and chronic disease states. |
Spontaneous clearance of hepatitis C virus in a patient co-infected with hepatitis C virus and human immunodeficiency virus: a case report. |
The effect of highly active antiretroviral therapy (HAART) on hepatitis C virus (HCV) infection remains unclear. Spontaneous HCV clearance with initiation of HAART in non-cirrhotic HCV patients co-infected with human immunodeficiency virus (HIV) has been reported. We describe an HIV/HCV patient with decompensated cirrhosis, who had spontaneous HCV clearance after an episode of elevated liver enzymes and a change in HAART regimen. His HCV RNA level remained undetectable for six months by quantitative and qualitative polymerase chain reaction (PCR) tests. The disappearance of HCV RNA may be due to a combination of host immune recovery, genetic polymorphism and direct effect of HAART against HCV. |
A potential treatment of non-alcoholic fatty liver disease with SIRT1 activators. |
Sirtuins (SIRTs) are members of the silent information regulator-2 family and act as nicotinamide adenine dinucleotide (NAD+)-dependent histone/protein deacetylases. The de-acetylation of proteins and histones results in an up- or down-regulation of gene transcription and protein function. In recent years, the regulatory action of the deacetylation activity of SIRT1 has been shown to have a positive impact on the pathophysiological mechanisms of nonalcoholic fatty liver disease (NAFLD). Among the effects of SIRT1 are: its healing activity on insulin sensitivity, thereby ameliorating glycemic regulation; its mimetic activity on calorie restriction; its antihyperlipidemic activity on lipid homeostasis via the liver, adipose tissues and skeletal muscles; its anti-inflammatory activities; its protective effects against cardiovascular events and endothelial dysfunction; its positive influence on autophagy, apoptosis and cancer; and finally, its anti-aging activity. The current approach for the treatment of NAFLD involves the treatment of etiological factors and recommendation of life-style changes including more physical activity and a low-calorie diet. However, there is no specific medical treatments for NAFLD. The therapeutic potential of SIRT1 activity in the treatment of NAFLD discovered in humans has been presented in this article. In this review, the potential effects of SIRT1 activation on NAFLD-related pathophysiological mechanisms and on the treatment of NAFLD are discussed. |
Pathophysiological and clinical approach to cirrhotic cardiomyopathy. |
Hyperdynamic circulation, systolic and diastolic left ventricular dysfunction and certain electrophysiological abnormalities have been associated with cirrhosis and known for a long time. These clinical features have been introduced as cirrhotic cardiomyopathy (CCM), which is characterized by blunted myocardial contractile responsiveness to physical, physiological and pharmacological stress. Importantly, cardiac dysfunction can be reversible and can improve due to effective medical treatment and also after liver transplantation. Echocardiography and electrocardiography are essential tools for recognizing the characteristic changes in the myocardial function and also the alterations in the electrophysiological properties of the heart. Laboratory markers are auxiliary modalities further aiding the establishment of the correct diagnosis. In this review, we aimed to collect the pathophysiological background and clinical characteristics of CCM with the intention of summarizing the current possibilities for the diagnosis establishment and treatment of this cardio-hepatic disorder. |
What non-alcoholic fatty liver disease has got to do with obstructive sleep apnoea syndrome and viceversa? |
Non-alcoholic fatty liver disease (NAFLD) and obstructive sleep apnoea syndrome (OSAS) are common conditions, frequently encountered in patients with obesity and/or metabolic syndrome. NAFLD and OSAS are complex diseases that involve an interaction of several intertwined factors. Several lines of evidence lend credence to an immune system derangement in these patients, i.e. the low grade chronic inflammation status, reckoned to be the most important factor in causing and maintaining these two illnesses. Furthermore, it is emphasized the main role of spleen involvement, as a novel mechanism. In this review the contribution of the visceral adiposity in both NAFLD and OSAS is stressed as well as the role of intermittent hypoxia. Finally, a post on the prevention of systemic inflammation is made. |