Improved Sleep Quality Is Associated with Reduced Insulin Resistance in Cancer Survivors Undertaking Circuit, Interval-Based Exercise.

PURPOSE: Cancer patients often experience poor sleep quality, typically induced by cancer-related treatments, a sedentary lifestyle, and psychological distress, leading to an increased risk of metabolic dysregulation such as obesity and insulin resistance. In this novel 16-week pilot study, we examined the effect of a circuit-based aerobic and resistance exercise intervention on self-reported sleep quality in breast, prostate, and colorectal cancer survivors and explored the association between changes in sleep quality and insulin resistance. METHODS: Survivors of breast, prostate or colorectal cancers who were sedentary, overweight or obese (BMI>25.0 kg/m2) were randomized to exercise (n=60) or usual care (n=30). The 16-week intervention included supervised moderate-vigorous aerobic (65-85% of VO2max) and resistance (65-85% of 1-repetition maximum) exercise performed in a circuit, interval fashion three times per week. Patient-reported sleep quality and insulin resistance were assessed at baseline and post-intervention using Pittsburgh Sleep Quality Index (PSQI) and Homeostasis Model of Assessment (HOMA-IR), respectively. Mean changes in PSQI score that are negative demonstrate improvements in sleep. Between-group differences were determined using repeated-measures analysis of variance. Associations between changes in PSQI and insulin resistance were computed using Pearson correlations. RESULTS: Participants were 63.2±10.8 years old, obese (87%), female (55%), and completed chemotherapy + radiation therapy (75%). Adherence to the intervention was 92% and the retention rate was 100%. Post-intervention, the PSQI global score improved significantly in the exercise group when compared to usual care (mean between-group difference, -2.7; 95% CI, -4.2 to -0.6). Change in PSQI was inversely associated with change in HOMA-IR (r=-0.91; p<0.01) among the exercise group. CONCLUSIONS: A circuit, interval-based aerobic and resistance exercise intervention improved patient-reported sleep quality in breast, prostate, and colorectal cancer survivors. Additionally, this exercise-induced improvement in sleep-quality may result in reduced insulin resistance.

High expression of CD7 on CD34+ cells is not linked to deletion of derivative chromosome 9 or lack of dendritic cells in chronic myeloid leukaemia.

OBJECTIVE: High expression of CD7 on CD34+ cells (>20 %) has been shown to be associated with inferior prognosis in chronic myeloid leukaemia (CML), but the reason has not been unravelled. We set out to investigate whether lack of dendritic cells or der(9)t(9;22)(q34;q11) deletions might be correlated with increased CD7 expression on CD34+ cells in CML. MATERIAL AND METHODS: We identified 43 patients in our cohort of CML patients in the first chronic phase in whom we were able to assess the expression of CD7 on CD34+ cells. der(9)t(9;22) deletions were evaluated by FISH (fluorescent in situ hybridization) analyses and the proportions of plasmacytoid and myeloid dendritic cells were assessed by flow cytometry. RESULTS: High and low expressions of CD7 on CD34+ cells were found in 19 and 24 patients, respectively. Two out of 20 patients examined had a der(9)t(9;22)(q34;11) deletion, one patient with high expression and one with low expression of CD7 on CD34+ cells. The proportions of plasmacytoid dendritic cells (PDCs) and myeloid dendritic cells (MDCs) were reduced in a majority of patients in our cohort, but no correlation was found between high or low expression of CD7 on CD34+ cells and the proportion of dendritic cells. CONCLUSIONS: A high proportion of CD34+CD7+ cells in patients with CML is not associated with der(9)t(9;22)(q34;q11) deletions. Nor did we find any correlation between CD7 expression on CD34+ cells and lack of dendritic cells. High expressions of CD7 on CD34+ cells and der(9)t(9;22)(q34;q11) deletions seem to be independent prognostic markers in CML.

Characterization of a-SiC(x):H thin films as an encapsulation material for integrated silicon based neural interface devices.

A fully integrated, wireless neural interface device is being developed to free patients from the restriction and risk of infection associated with a transcutaneous wired connection. This device requires a hermetic, biocompatible encapsulation layer at the interface between the device and the neural tissue to maintain long-term recording/stimulating performance of the device. Hydrogenated amorphous silicon carbide (a-SiC(x):H) films deposited by a plasma enhanced chemical vapor deposition using SiH(4), CH(4), and H(2) precursors were investigated as the encapsulation layer for such device. Si-C bond density, measured by Fourier transform infrared absorption spectrometer, suggests that deposition conditions with increased hydrogen dilution, increased temperature, and low silane flow typically result in increase of Si-C bond density. From the variable angle spectroscopic ellipsometry measurement, no dissolution of a-SiC(x):H was observed during soaking tests in 90°C phosphate buffered saline. Conformal coating of the a-SiC(x):H in Utah electrode array was observed by scanning electron microscope. Electrical properties were studied by impedance spectroscopy to investigate the performance of a-SiC(x):H as an encapsulation layer, and the results showed long term stability of the material.

Treatment results in localized primary gastric lymphoma: data of patients registered within the German multicenter study (GIT NHL 02/96).

PURPOSE: In the prospective study 02/96 on primary GI lymphoma, we have collected data on histology, clinical features, and treatment results. In particular, in stages I and II localized primary gastric lymphoma (PGL), our objectives were to reduce treatment intensity and to confirm our hypothesis from study 01/92, which maintained that an organ-preserving approach is not inferior to primary surgery. PATIENTS AND METHODS: Patients receiving radiotherapy and/or chemotherapy were stratified for histologic grade, stage, and whether surgery had been carried out or not (as decided by each participating center). Patients with aggressive PGL received six cycles of CHOP-14 (cyclophosphamide, doxorubicin, vincristine, and prednisone) followed by involved-field radiotherapy (40 Gy). Patients with indolent PGL (including patients experiencing treatment failure with antibiotic therapy for Helicobacter pylori) were treated with extended-field radiotherapy. The volume depended on stage. The irradiation dose was 30 Gy, followed by a boost of 10 Gy (the latter omitted after complete resection) to the tumor region. RESULTS: Seven hundred forty-seven patients were accrued. Of these patients, 393 with localized PGL were treated with radiotherapy and/or chemotherapy only or additional surgery between December 1996 and December 2003. The survival rate at 42 months for patients treated with surgery was 86% compared with 91.0% for patients without surgery. CONCLUSION: In this nonrandomized study (02/96), we reproduced the previous results of study 01/92 showing no disadvantage for an organ-preserving treatment. Therefore, primary stomach resection should be questioned.

Microdevices for separation, accumulation, and analysis of biological micro- and nanoparticles.

Microfabrication and performance of a novel microsystem for separation, accumulation and analysis of biological micro- and nanoparticles is reported. Versatile chip functions based on dielectrophoresis and microfluidics were integrated to isolate particles from complex sample solutions such as serum. A bead-based assay for virus detection is proposed. Separation of micro- and sub-mum beads employing dielectrophoretic deflector and bandpass structures is demonstrated. Individual antibody coated beads with hepatitis A virus bound to their surface were trapped by negative dielectrophoresis in a field cage and analysed by fluorescence microscopy.

Accelerated radiochemotherapy in pancreatic cancer is not necessarily related to a pathologic pancreatic function decline in the early period.

PURPOSE: To evaluate the functional effects of ionizing radiation in patients with unresectable pancreatic cancer in the early period after accelerated radiochemotherapy (ART). METHODS AND MATERIALS: To analyze the exocrine component, the amino acid consumption test and fecal elastase 1 were performed in 13 patients immediately before and 4-8 weeks after ART. Pancreatic duct morphology was evaluated before therapy. Weight loss and clinical steatorrhea were recorded. Endocrine parameters were examined according to standardized criteria. RESULTS: The relative change of the amino acid consumption test results and the median elastase concentration was 41.2% and 56.4%, respectively. Five patients still had normal test results after ART and 5 patients developed pathologic values. The median relative weight loss of the total body weight was 7.7% +/- 4.5%. No steatorrhea occurred. Of the 5 patients with normal values, 3 had a mean organ dose of <40 Gy. Of the 5 patients with pathologic values, 4 had a mean organ dose of >41 Gy. The endocrine function measurements remained unchanged. CONCLUSION: Although a nominal reduction of exocrine function parameters occurred in most patients, ART was not necessarily related to a pathologic level in the early period. Diabetes was not established. The functional impairment that was existent in the patient population presumably contributed to the weight loss. Pancreatic enzyme preparations may also play a role in maintaining an anabolic state during and after radiochemotherapy.

Conformal therapy for pancreatic cancer: variation of organ position due to gastrointestinal distention--implications for treatment planning.

PURPOSE: To quantify nonrespiratory organ motion in the pancreatic region and its effect on clinical target volume. MATERIALS AND METHODS: Three-dimensional translations of the geometric centers of the volumes of interest--pancreatic head, body, and tail; left and right kidney; and the superior mesenteric artery--were measured in 20 patients by analyzing three spiral computed tomographic (CT) protocols performed at static exhalation and representing differential gastrointestinal distention. Wilcoxon test for paired differences was applied to determine statistical significance (P <.05). Spearman rank correlation coefficients were calculated between combinations of statistically significant translations. With the assumption that the organ positions were represented by a three-dimensional Gaussian distribution that occurs during treatment, clinical target volume expansions were calculated to account for organ motion and a typical setup error. RESULTS: Significant translations of the volume of interest were observed. The most mobile parts of the target organs were the pancreatic tail (P =.001) and the superior mesenteric artery (P =.01). Larger variations from the mean in the planning CT protocol in which negative contrast material was used usually resulted in a slightly larger clinical target volume expansion. CONCLUSION: Our data may provide a basis for further studies of organ motion and ways of modifying treatment margins.

Genetic relatedness of Cuban HAV wild-type isolates.

Knowledge of the nucleotide sequence in the region of the putative VP1/2A junction of the Hepatitis A virus (HAV) genome has enabled differentiation of HAV strains and their classification into seven genotypes, in some of which sub-genotypes A and B can be defined. A 168 base segment encompassing the putative VP1/2A junction of 27 clinical wild-type isolates of HAV from Cuba was amplified by reverse transcriptase-polymerase chain reaction and then sequenced. The Cuban isolates are clustered within sub-genotype IA. A single amino acid substitution, which does not correspond with any of the reported changes for other strains, was observed. A new sub-genotype variant is therefore postulated. This study provides new data on the genetic relatedness of HAVs from Cuba and on the distribution of sub-genotype IA in the Caribbean area.

Influence of the 5' noncoding region of hepatitis A virus strain GBM on its growth in different cell lines.

Previous sequence analysis of consecutive passages of the hepatitis A virus (HAV) strain GBM/WT in human embryonic kidney cells (HEK cells), human embryonic lung fibroblasts (HFS cells) and in FRhK-4 cells (foetal rhesus monkey kidney cells) pointed to a host cell dependent cell culture adaptation of GBM/WT in HFS cells involving mutations in the 5' noncoding region (5'NCR). Multiple nucleotide changes occurred in the 5'NCR of the GBM genome after the cell line used for virus passage was changed from HEK cells to HFS cells. In contrast, no mutations in the 5'NCR occurred during the first 20 passages of GBM/WT in FRhK-4 cells. In order to analyse the influence of the 5'NCR on host cell specific adaptation of HAV strain GBM in different cell cultures, GBM/HM175 chimeras were constructed which contained 5'NCRs from different GBM variants by replacing the 5'NCR of the infectious clone pHAV/7. Parallel transfection assays in FRhK-4 and HFS cells, performed with transcripts from the chimeric GBM/HM175 constructs, showed that the 5'NCR of the GBM variant GBM/HFS is essential for virus growth in HFS cells. The GBM/HM175 chimeric RNA, which contained the 5'NCR of GBM/HFS, exclusively, was able to produce infectious virus after transfection of HFS cells. The growth of the different GBM/HM175 chimeras in FRhK-4 cells, in contrast, did not seem to be strongly influenced by a specific sequence of the 5'NCR.

Molecular epidemiology of an outbreak of hepatitis A in Italy.

The relationship of hepatitis A virus (HAV) isolates associated with an outbreak in Genoa, Italy, in 1993 was examined using direct sequencing of amplicons derived by antigen capture PCR (AC/PCR) from faecal samples of the infected persons. Forty samples recovered from 38 primary and two secondary cases were examined. The latter were household contacts of the primary cases. In addition, faecal material of 2 unrelated persons infected simultaneously with hepatitis A in Genoa were tested. The PCR products derived from the P1/P2 junction of the HAV genome were analysed. A 100% nucleotide identity was detected between the viral isolates originating from the primary as well as the secondary cases. The viral isolates recovered from the faecal samples of the two unrelated cases differed from the virus causing the outbreak as well as from each other. These results indicate that a single HAV strain caused the outbreak. The virus might have been transmitted by ingestion of contaminated food or water since all hepatitis A infected employees of the factory had eaten in the same canteen. Definitions of HAV genotypes are based on numerous genetic comparisons of different strains. The sequence comparison of the investigated isolates with published HAV sequences of the P1/P2 genome region revealed that the virus associated with the outbreak belongs to HAV subgenotype IA, whereas the strains recovered from the viral isolates of the unrelated cases belong to subgenotype IB.

Hepatitis A virus and polymerase chain reaction amplification: methodology and results.

Hepatitis A infection among patients receiving solvent/detergent inactivated factor VIII preparations in various locations in Europe have been documented recently. In investigations in Italy, Germany and Ireland, polymerase chain reaction (PCR) amplification was used to detect hepatitis A virus in frozen plasma pools, purified factor VIII, patient sera and samples from animal transmission studies; nucleic acid sequencing was used to clarify and identify the virus responsible based upon genotype analysis. Unique virus strains were found among the cases in Italy and Germany, and identical virus sequences were also found in some factor VIII lots. However, with the exception of the Italian investigation, lack of appropriate samples have precluded the identification of virus in these outbreaks. In addition, animal infectivity studies have not been successful in demonstrating infectivity under laboratory conditions. We discuss the limitations of PCR amplification with respect to detecting virus within these situations, and the necessity for the corresponding epidemiologic investigations.

Mutational events in consecutive passages of hepatitis A virus strain GBM during cell culture adaptation.

In order to study the adaptation of hepatitis A virus (HAV) in cell culture, we examined the mutational events of the genome in early passages of HAV strain GBM propagated either in FRhK-4 cells (fetal rhesus monkey kidney-derived) or in human embryonic kidney (HEK) and human embryonic fibroblast cells (HFS) in relation to their growth characteristics. Sequence analysis of the nucleotide region encoding 2B, 2C, and the beginning of 3A as well as the nucleotide region encompassing the 5' noncoding region (5'NCR) of the genome was performed on consecutive virus passages after amplification of the viral RNA from the cell culture supernatant by antigen-capture PCR. By the 2nd passage of the GBM variants cultured in FRhK-4 or in HEK cells we found a mutation at nucleotide position 3889 (2B coding region) which results in an amino acid substitution from alanine to valine. Further mutations present in the 2B/2C region of the cell culture-adapted GBM variants differ from each other and occur after the 10th or even the 40th virus passage. Another early change, an in frame deletion of nine nucleotides in the 3A region, appeared in the 5th virus passage only in GBM cultured on FRhK-4 cells. This genome region showed different mutations in the virus passages on HEK and HFS cells. The 5'NCR of the cell culture-adapted GBM variants, in contrast, did not show any mutations before the 8th virus passage. The faster and more efficient growth of the HAV strain GBM during successive propagation on cell cultures seems to correlate with the appearance of mutations in the investigated genome regions.

Processing of truncated mouse or human rRNA transcribed from ribosomal minigenes transfected into mouse cells.

The processing of pre-rRNA in eukaryotic cells involves a complex pattern of nucleolytic reactions taking place in preribosomes with the participation of several nonribosomal proteins and small nuclear RNAs. The mechanism of these reactions remains largely unknown, mainly because of the absence of faithful in vitro assays for most processing steps. We have developed a pre-rRNA processing system using the transient expression of ribosomal minigenes transfected into cultured mouse cells. Truncated mouse or human rRNA genes are faithfully transcribed under the control of mouse promoter and terminator signals. The fate of these transcripts is analyzed by the use of reporter sequences flanking the rRNA gene inserts. Both mouse and human transcripts, containing the 3' end of 18S rRNA-encoding DNA (rDNA), internal transcribed spacer (ITS) 1, 5.8S rDNA, ITS 2, and the 5' end of 28S rDNA, are processed predominantly to molecules coterminal with the natural mature rRNAs plus minor products corresponding to cleavages within ITS 1 and ITS 2. To delineate cis-acting signals in pre-rRNA processing, we studied series of more truncated human-mouse minigenes. A faithful processing at the 18S rRNA/ITS 1 junction can be observed with transcripts containing only the 60 3'-terminal nucleotides of 18S rRNA and the 533 proximal nucleotides of ITS 1. However, further truncation of 18S rRNA (to 8 nucleotides) or of ITS 1 (to 48 nucleotides) abolishes the cleavage of the transcript. Processing at the ITS 2/28S rRNA junction is observed with truncated transcripts lacking the 5.8S rRNA plus a major part of ITS 2 and containing only 502 nucleotides of 28S rRNA. However, further truncation of the 28S rRNA segment to 217 nucleotides abolishes processing. Minigene transcripts containing most internal sequences of either ITS 1 or ITS 2, but devoid of ITS/mature rRNA junctions, are not processed, suggesting that the cleavages in vivo within either ITS segment are dependent on the presence in cis of mature rRNA sequences. These results show that the major cis signals for pre-rRNA processing at the 18S rRNA/ITS 1 or the ITS2/28S rRNA junction involve solely a limited critical length of the respective mature rRNA and adjacent spacer sequences.

Sequence variability of hepatitis A virus and factor VIII associated hepatitis A infections in hemophilia patients in Europe. An update.

Outbreaks and sporadic cases of hepatitis A have been observed in 4 European countries in hemophilia patients receiving factor VIII preparations. PCR amplification of potential hepatitis A virus (HAV) nucleic acid present in plasma pools, purified factor VIII and acute-phase sera from infected individuals has been performed and the nucleic acid sequence determined for those samples that resulted in a positive PCR product. HAV sequences were detected in the serum of 2 German patients, but not in the factor VIII lots administered to these individuals. Screening of plasma pools and the corresponding 5 lots of factor VIII associated with the outbreak in Ireland did not reveal any HAV sequences. In contrast, a study of samples from Italy detected HAV sequences in 5 of 12 lots and in 2 hemophilia patients who developed hepatitis A. These data suggest that implicated factor VIII preparations might have been involved in the outbreaks of HAV infection among Italian hemophiliacs. However, no molecular evidence was obtained for a similar association in Germany or Ireland. The preliminary data from these two investigations must be verified by animal inoculation studies and supported by epidemiologic analysis.

Detection of hepatitis A virus in a factor VIII preparation by antigen capture/PCR.

The antigen capture/PCR (AC/PCR) has been applied in the analysis of various factor VIII preparations, which were suspected to be contaminated with hepatitis A virus (HAV). AC/PCR involves capturing the antigen, i.e. the intact virus particles, by binding to the HAV monoclonal antibody mAb 7e7, reverse transcription of the viral RNA and amplification of the cDNA with HAV-specific primer pairs. The PCR analysis of one factor VIII concentrate yielded an HAV-specific DNA product, which could be confirmed by Southern blot analysis. The HAV strain recovered by AC/PCR from this factor VIII concentrate could be classified into genotype III after solid-phase sequencing of the product and comparison with the consensus sequences for the known HAV genotypes. Analysis of the sera from 3 haemophiliacs treated with this batch has not resulted in a reliable product. However, the results obtained indicate that HAV can be detected in purified factor VIII preparations by AC/PCR.

Nucleotide sequence of wild-type hepatitis A virus GBM in comparison with two cell culture-adapted variants.

In order to study cell tropism and attenuation of hepatitis A virus (HAV), the genome of HAV wild-type GBM and two cell culture-adapted variants, GBM/FRhK and GBM/HFS, were cloned and sequenced after amplification by reverse transcriptase-PCR. During virus cultivation, the HAV variant GBM/FRhK had a strict host range for FRhK-4 cells, in contrast to GBM/HFS, which can be grown in HFS and FRhK-4 cells. The HAV variant GBM/HFS was shown to be attenuated when inoculated into chimpanzees (B. Flehmig, R. F. Mauler, G. Noll, E. Weinmann, and J. P. Gregerson, p. 87-90, in A. Zuckerman, ed., Viral Hepatitis and Liver Disease, 1988). On the basis of this biological background, the comparison of the nucleotide sequences of these three HAV GBM variants should elucidate differences which may be of importance for cell tropism and attenuation. The comparison of the genome between the GBM wild type and HAV wild types HM175 (J. I. Cohen, J. R. Ticehurst, R. H. Purcell, A. Buckler-White, and B. M. Baroudy, J. Virol. 61:50-59, 1987) and HAV-LA (R. Najarian, O. Caput, W. Gee, S. J. Potter, A. Renard, J. Merryweather, G. Van Nest, and D. Dina, Proc. Natl. Acad. Sci. USA 82:2627-2631, 1985) showed a 92 to 96.3% identity, whereas the identity was 99.3 to 99.6% between the GBM variants. Nucleotide differences between the wild-type and the cell culture-adapted variants, which were identical in both cell culture-adapted GBM variants, were localized in the 5' noncoding region; in 2B, 3B, and 3D; and in the 3' noncoding region. Our result concerning the 2B/2C region confirms a mutation at position 3889 (C-->T, alanine to valine), which had been shown to be of importance for cell culture adaptation (S. U. Emerson, C. McRill, B. Rosenblum, S. M. Feinstone, and R. H. Purcell, J. Virol. 65:4882-4886, 1991; S. U. Emerson, Y. K. Huang, C. McRill, M. Lewis, and R. H. Purcell, J. Virol. 66:650-654, 1992), whereas other mutations differ from published HAV sequence data and may be cell specific. Further comparison of the two cell culture-adapted GBM variants showed cell-specific mutations resulting in deletions of six amino acids in the VP1 region and three amino acids in the 3A region of the GBM variant GBM/FRhK.