Early Protective Effects of Iloprost after Experimental Spinal Cord Ischemia in Rabbits

The potential role of Iloprost, a stable analogue of prostocyclin, in treating spinal cord ischemia was investigated in rabbits subjected to aortic occlusion for 15 minutes. Ten adult rabbits weighing 2-2.5 kg received an intravenous infusion of saline (SF) as a control group and 14 rabbits received an intravenous infusion of Iloprost, 25 microg/kg/h. Iloprost infusion was started immediately after clamping of the aorta and continued 60 minutes thereafter. Cortical somatosensorial evoked potentials (CSEP) were recorded during the pre-ischemic period as a baseline and post-ischemic readings were taken at 15, 30 and 60 minutes. There was no statistically significant difference between CSEP of the saline and Iloprost treated groups (p < 0.05). All animals were examined neurologically by using a modification of Tarlov scale and all subjects were then deeply anesthetized and their spinal cords were removed for light and electron microscopic examinations at 24 h after spinal cord ischemia. In order to obtain an accurate comparison of ultrastructural changes between saline treated and Iloprost treated groups, a grading scale was performed. The light microscopic and ultrastructural analysis of the Iloprost treated group revealed that there was moderate protection of the myelin and axons and edema was attenuated. Findings of this study suggest that Iloprost exerts a protective effect on spinal cord ischemia. However, further studies are needed to reveal possible mechanisms of protection provided by Iloprost.     

A prospective cohort study on the prediction of the diagnosis-to-delivery time in preeclamptic pregnancies: should the sFlt-1/PlGF ratio be added to routine evaluations?

Purpose

To analyze the clinical and laboratory factors that potentially affect the diagnosis-to-delivery time in preeclamptic pregnancies.

Methods

In this cross-sectional study, we followed 24 early onset preeclampsia (E-PE) and 26 late-onset preeclampsia (L-PE) cases. Maternal serum samples were obtained at the time of diagnosis and stored at ??80 °C until ELISA analysis for soluble fms-like tyrosine kinase-1 (SFlt-1) and placental growth factor (PlGF) levels.

Results

The median follow-up duration was 68 (1–339) h in the E-PE group and 330 (7–1344) h in the L-PE group. Maternal mean arterial pressure (MAP) at hospitalization was the strongest variable, and the sFlt-1/PlGF ratio added significantly to the Cox regression model. In the E-PE cases, the median sFlt-1/PlGF ratio was significantly higher in the subgroup with a follow-up duration?>?48 h than in the subgroup of cases with a follow-up duration?≤?48 h (5109 vs. 2080; p?=?0.038), and none of the seven cases with an sFlt-1/PlGF ratio?≥?75th percentile delivered during the first 48 h. Neither the 24-h proteinuria nor the gestational age at diagnosis added to the predictive power of the MAP at hospitalization.

Conclusion

Incorporation of the sFlt-1/PlGF ratio to the routine evaluation of preeclamptic pregnancies may help in the prediction of progression and management planning.

     

Sympathetic overactivity in patients with left ventricular aneurysm in early period after anterior myocardial infarction: does sympathetic activity predict aneurysm formation?

Left ventricular aneurysm (LVA) is an important complication of acute transmural myocardial infarction (MI) that bears great clinical significance because of high mortality. Heart rate variability (HRV) analysis is extensively used to evaluate autonomic modulation of sinus node and to identify patients at risk for an increased cardiac mortality. In this study, the authors evaluated HRV in patients with LVA in the early period after acute anterior wall MI. They compared 18 patients (7 men, 11 women, with an average age of 56.1 +/-8.2 years) with LVA and 46 patients (34 men, 12 women, with an average age of 56.4 +/-5.9 years) without LVA. Mean heart rate, low frequency (LF) and low-frequency/high-frequency (LF/HF) ratio were significantly higher and standard deviation of normal-to-normal RR intervals (SDNN), root mean square of successive differences (RMSSD), number of NN intervals that differed by more than 50 ms from adjacent interval divided by the total number of all NN intervals (PNN50), and HF were lower in the patients with LVA. A SDNN <78 ms separated the patients with aneurysm from those without aneurysm with a sensitivity of 78%, specificity of 83%, positive predictive accuracy of 79%; a LF/HF ratio >2.4 with a sensitivity of 92%, specificity of 88%, and positive predictive accuracy of 92%. Single-vessel disease increased the left ventricular aneurysm formation by 5.1 fold, total left anterior descending artery (LAD) occlusion by 3.1 fold, mean heart rate >75 beats/minute by 2.3 fold, SDNN <78 ms by 7.9 fold, and LF/HF ratio >2.4 by 12.9 fold, but well-developed collaterals decreased the aneurysm formation by 4.4 fold. As a result, HRV analysis supplies parameters with high predictive value for LVA formation in the early period after acute anterior MI. The higher sympathetic activity and reduced heart rate variability may be associated with a higher incidence of complications such as ventricular arrhythmias and increased mortality in patients with LVA.     

The global burden of headache in children and adolescents – developing a questionnaire and methodology for a global study

Background

Burden of headache has been assessed in adults in countries worldwide, and is high, but data for children and adolescents are sparse. The objectives of this study were o develop a questionnaire and methodology for the global estimation of burden of headache in children and adolescents, to test these in use and to present preliminary data.

Methods

We designed structured questionnaires for mediated-group self-administration in schools by children aged 6-11 years and adolescents aged 12-17 years. In two pilot studies, we offered the questionnaires to pupils in Vienna and Istanbul. We performed face-to-face interviews in a randomly selected subsample of 199 pupils to validate the headache diagnostic questions.

Results

Data were collected from 1,202 pupils (mean 13.9 ± 2.4 years; 621 female, 581 male). The participation rate was 81.1% in Istanbul, 67.2% in Vienna. The questionnaire proved acceptable: ≤5% of participants disagreed partially or totally with its length, comprehensibility or simplicity. The sensitivity, specificity, positive and negative predictive values ranged between 0.71 and 0.76 for migraine and between 0.61 and 0.85 for tension-type headache (TTH). Cronbach’s alpha was 0.83. The 1-year prevalence of headache was 89.3%, of migraine 39.3% and of TTH 37.9%. The prevalence of headache on ≥15 days/month was 4.5%. One fifth (20.7%) of pupils with headache lost ≥1 day of school during the preceding 4 weeks and nearly half (48.8%) reported ≥1 day when they could not do activities they had wanted to. The vast majority of pupils with headache experienced difficulties in coping with headache and in concentrating during headache. Quality of life was poorer in pupils with headache than in those without.

Conclusion

These pilot studies demonstrate the usefulness of the questionnaires and feasibility of the methodology for assessing the global burden of headache in children and adolescents, and predict substantial impact of headache in these age groups.     

Exclusive TCRVbeta chain usage of ex vivo generated minor Histocompatibility antigen HA-1 specific cytotoxic T cells: implications for monitoring of immunotherapy of leukemia by TCRBV spectratyping

Tissue expression of minor Histocompatibility antigens HA-1 and HA-2 is limited to the hematopoietic system. Therefore, ex vivo generated HA-1/HA-2 specific cytotoxic T lymphocytes (CTLs) can be applied for adoptive immunotherapy of relapsed leukemia after HLA-matched HA-1/HA-2 mismatched stem cell transplantation. Here we used T cell receptor beta variable chain (TCRBV) spectratyping and/or TCRBV sequencing to monitor the specific TCR usage in eleven HA-1/HA-2 CTLs that were induced ex vivo with peptide pulsed dendritic cells. The HA-2 induced CTLs used different TCRBV. In contrast, the development of HA-1 specific CTLs coincided with prominent skewing of TCRBV7 spectratypes. Sequencing of the TCRBV7 specific PCR products used by these ex vivo generated HA-1 CTLs revealed the exclusive usage of TCRBV7-9*03, identical to the TCRBV used by HA-1 specific CTLs induced in vivo after stem cell transplantation. Thus, monitoring of immunotherapy with HA-1 specific CTLs is now also feasible by TCRBV spectratyping.     

Generation of minor histocompatibility antigen HA-1-specific cytotoxic T cells restricted by nonself HLA molecules: a potential strategy to treat relapsed leukemia after HLA-mismatched stem cell transplantation

Successful stem cell transplantation (SCT) across HLA barriers can be performed with cord blood, megadoses of stem cells, or with nonmyeloablative conditioning strategies. Because the HLA-mismatched transplants are often T-cell depleted, leukemia relapse rates are high. Treatment of relapsed leukemia after HLA-mismatched SCT is difficult. A novel potential strategy to treat relapsed leukemia after HLA-mismatched SCT is the use of patients' mismatched HLA molecules as antigen-presenting molecules to generate hematopoietic system-specific cytotoxic T cells (CTLs) from the stem cell donor. Adoptive transfer of these hematopoietic system-specific CTLs that are restricted by nonself HLA molecules may eliminate leukemia without affecting the patient's nonhematopoietic cells or donor hematopoietic cells. We investigated the feasibility of this strategy using the hematopoietic system-specific minor histocompatibility antigen HA-1, which is known to induce HLA-A2-restricted CTLs. HLA-A2(-) peripheral blood mononuclear cells were stimulated with HLA-A2(+) T2 cells pulsed with synthetic HA-1 peptide or with dendritic cells transduced with the HA-1 cDNA. Tetrameric HLA-A2/HA-1 peptide complexes were used to monitor and enrich HA-1-specific CTLs. In the alloreactive cultures, HA-1-specific CTLs were enriched up to 7% by 3 rounds of antigen-specific stimulations and up to 87% by fluorescence-activated cell sorting of tetramer-positive T cells. The HA-1-specific CTLs showed specific lysis of the relevant target cells, including leukemic cells. Because the polyclonal CTL cultures also contained natural killer cells and allo-HLA-A2-specific CTLs, CTL clones were generated that showed the expected HA-1 specificity only. Thus, HA-1-specific CTLs restricted by nonself HLA-A2 molecules can be generated in an HLA-A2-mismatched setting.     

Anti-HCV positivity in sexual partners and offspring of patient with chronic hepatitis C

We investigated the seroprevalence of HCV in stable sexual partners and offspring of chronic hepatitis C patients, and aimed to determine the risk factors involved. 191 anti-HCV and HCV RNA positive subjects who coinhabited with their spouse and/or offspring were included. Risk factors of index cases for disease transmission, liver biopsy results, anti-HCV and HCV-RNA in spouses and/or offspring were evaluated. Together with index cases, a total of 404 family members including 174 stable sexual partners and 230 offspring were included. The most common risk factors among index cases were dental procedures (73.8%), history of surgery (64.9%), and blood transfusions (24.1%). Anti-HCV positivity was established in 11 (2.7%) of the total 404 family contacts--6 sexual partners and 5 offspring. HCV seropositivity was significantly higher in the spouses of index cases with severe hepatitis C compared to those with mild to moderate hepatitis C (p=0.008), but there was no statistically significant correlation between the severity of liver disease in index cases and anti-HCV positivity in their offspring. In conclusion, anti-HCV seropositivity in the spouses and children of patients who are HCV-RNA positive HCV carriers does not appear to be higher than the HCV seroprevalence in our country.     

Lenalidomide maintenance after nonmyeloablative allogeneic stem cell transplantation in multiple myeloma is not feasible: results of the HOVON 76 Trial

To improve the outcome of allogeneic stem cell transplantation (allo-SCT) in multiple myeloma as part of first-line treatment, we prospectively investigated the feasibility and efficacy of lenalidomide maintenance. Patients started maintenance 1 to 6 months after nonmyeloablative allo-SCT. Lenalidomide was dosed 10 mg on days 1 to 21 of a 28-day schedule for a total of 24 cycles. Peripheral blood samples were taken to evaluate immune modulating effects. Thirty-five eligible patients were enrolled, and 30 started with lenalidomide. After 2 cycles, 14 patients (47%) had to stop treatment, mainly because of the development of acute graft versus host disease (GVHD). In total, 13 patients (43%) stopped treatment because of development of GVHD, 5 patients (17%) because of other adverse events, and 5 patients (17%) because of progression. Responses improved in 37% of patients, and the estimated 1-year progression-free survival from start of maintenance was 69% (90% confidence interval, 53%-81%). Lenalidomide increased the frequency of human leukocyte antigen-DR(+) T cells and regulatory T cells, without correlation with clinical parameters. In conclusion, lenalidomide maintenance 10 mg daily after nonmyeloablative allo-SCT with unmanipulated graft in multiple myeloma patients is not feasible, mainly because of the induction of acute GVHD. This trial was registered at www.trialregister.nl as #NTR1645.