The signature of a T-cell response to KSHV persists across space and time in individuals with epidemic and endemic KS from Uganda.

Inadequate T-cell control of Kaposi sarcoma-associated herpesvirus (KSHV) infection predisposes to development of Kaposi sarcoma (KS), but little is known about the T-cell response to KSHV. Postulating that KS tumors contain abundant KSHV-specific T-cells, we performed transcriptional profiling and T-cell receptor (TCR) repertoire analysis of tumor biopsies from 144 Ugandan adults with KS. We show that CD8+ T-cells and M2-polarized macrophages dominate the tumor micro-environment (TME). The TCR repertoire of KS tumor infiltrating lymphocytes (TIL) is shared across non-contiguous tumors and persists across time. Clusters of T-cells with predicted shared specificity for uncharacterized antigens, potentially encoded by KSHV, comprise ~25% of KS TIL, and are shared across tumors from different time points and individuals. Single-cell RNA-sequencing of blood identifies a non-proliferating effector memory phenotype and captured the TCRs in 14,698 putative KSHV-specific T-cells. These results suggest that a polyspecific KSHV-specific T-cell response inhibited by M2 macrophages exists within the KS TME, and provide a foundation for studies to define its specificity at a large scale.

Mavorixafor, an Orally Bioavailable CXCR4 Antagonist, Increases Immune Cell Infiltration and Inflammatory Status of Tumor Microenvironment in Patients with Melanoma.

Purpose: Mavorixafor is an oral, selective inhibitor of the CXCR4 chemokine receptor that modulates immune cell trafficking. A biomarker-driven phase Ib study (NCT02823405) was conducted in 16 patients with melanoma to investigate the hypothesis that mavorixafor favorably modulates immune cell profiles in the tumor microenvironment (TME) and to evaluate the safety of mavorixafor alone and in combination with pembrolizumab. Experimental Design: Serial biopsies of melanoma lesions were assessed after 3 weeks of mavorixafor monotherapy and after 6 weeks of combination treatment for immune cell markers by NanoString analysis for gene expression and by multiplexed immunofluorescent staining for in situ protein expression. Serum samples taken at biopsy timepoints were evaluated for key chemokine and cytokine alterations using the Myriad Rules Based Medicine multiplex immunoassays. Results: Within the TME, mavorixafor alone increased CD8+ T-cell infiltration, granzyme B signal, antigen presentation machinery, and both tumor inflammatory signature (TIS) and IFNγ gene expression signature scores. Increases in the key serum cytokines CXCL9 and CXCL10 were further enhanced when mavorixafor was combined with pembrolizumab. Adverse events (AE), as assessed by the investigator according to NCI Common Terminology Criteria for Adverse Events (v4.03), related to either mavorixafor or pembrolizumab (≥15%) were diarrhea, fatigue, maculopapular rash, and dry eye. Reported AEs were all ≤ grade 3. Conclusion/Discussion: Treatment with single-agent mavorixafor resulted in enhanced immune cell infiltration and activation in the TME, leading to increases in TIS and IFNγ gene signatures. Mavorixafor as a single agent, and in combination with pembrolizumab, has an acceptable safety profile. These data support further investigation of the use of mavorixafor for patients unresponsive to checkpoint inhibitors. Significance: Despite survival improvements in patients with melanoma treated with checkpoint inhibitor therapy, a significant unmet medical need exists for therapies that enhance effectiveness. We propose that mavorixafor sensitizes the melanoma tumor microenvironment and enhances the activity of checkpoint inhibitors, and thereby may translate to a promising treatment for broader patient populations.

Scope of practice distinctions based on primary work setting for genetic counselors in assisted reproductive technologies.

OBJECTIVE: To elucidate the tasks within various work settings that assisted reproductive technologies (ART) genetic counselors believe to be within their scope of practice. DESIGN: A survey was constructed and administered to genetic counselors who practice in the field of ART. SETTING: Genetic counselors were asked to self-identify with a primary ART work setting: genetic testing laboratory (preimplantation genetic testing, carrier screening, or both), in vitro fertilization clinic, gamete donor agency, telegenetic practice (either private practice or telemedicine company), or other. PATIENTS: N/A. INTERVENTIONS: N/A. MAIN OUTCOME MEASURES: The number of years of practice in ART, tasks performed within various ART work settings representing the reality or the ideal, and perception of understanding of the scope of practice by nongenetics colleagues. RESULTS: The majority of respondents reported <10 years of experience in this field. There were differences in what was considered the scope of practice among the various work settings. ART genetic counselors believed that their scope of practice was not well understood by their nongenetics colleagues. They also reported differences between the actual duties performed and what they ideally believed would be within their job function. CONCLUSIONS: The genetic counseling specialty of ART is a new work setting for genetic counselors. There is a need for education regarding the various roles of genetic counselors in ART. Better definition of the appropriate duties for genetic counselors in the various ART work settings is needed to foster effective working relationships with their nongenetics colleagues and optimize patient care.

Implications of a Retrospective Study on Weight-Based Risk for Post-Tonsillectomy Pain in Children.

PURPOSE: The purpose was to examine if children experience weight-based risks for post-tonsillectomy pain (PTP) in the postanesthesia care unit (PACU). DESIGN: This retrospective correlational cohort design included a sample of 180 children between the ages of 4 to 12 years who had tonsillectomy and adenoidectomy or tonsillectomy before August 2016; half were obese (OB) or overweight (OW). METHODS: The sample was obtained from children who had surgery at a large pediatric hospital with an attached outpatient surgical center in North Texas. Children were defined as either OB and OW or non-OB and non-OW based on a cutoff of standardized body mass index z scores of 85th percentile and greater per the National Center for Health Statistics. Pain scores were obtained in the PACU after surgery. Early PTP was defined as the most severe pain experienced by a child in the first 15 minutes after extubation. Prolonged PTP was sustained and uncontrolled pain in the PACU. FINDINGS: OB and OW status did not increase the likelihood of experiencing early PTP when examined by multiple logistic regression controlling for covariates (adjusted odds ratio, 1.391; P = .369). OB and OW status was associated with longer episodes of prolonged PTP (rs[178] = 0.16; P = .03). OB and OW children were more likely to experience prolonged PTP in the PACU (χ2[1] = 8.353; P = .004), with these children experiencing an average PTP period twice as long as their peers. CONCLUSIONS: OB and OW children did experience risk for prolonged PTP, averaging sustained pain for approximately twice as long as other children. The increased risk for prolonged PTP in OB and OW children occurred despite well-managed early PTP with rates that matched those of their peers. No weight-based risk for early PTP was observed. Further research is needed in the area of PTP management in OB and OW children.

Framework to Explain the Progression of Pain in Obese or Overweight Children Undergoing Tonsillectomy.

An estimated 100,000 obese (OB) and overweight (OW) children undergo tonsillectomy each year in the United States. Pain management in this population is particularly challenging because of weight-based dosing, clinician fears, potential for airway obstruction, and genetic differences. A framework is proposed to explain factors involved in the post-tonsillectomy pain (PTP) experience in OB and OW children. The tonsillectomy, the body's inflammatory state, and mechanical stressors comprise influencing factors in PTP progression. Clinician-delivered medication doses, genetic variants of drug metabolism, and soothing factors serve as mediating factors in the progression of PTP. Postanesthesia care unit (PACU) nurses may use this framework to better understand PTP progression in OB and OW children. PACU nurses may manipulate certain mediating factors discussed in this framework to moderate PTP progression in OB and OW children. Researchers may use this framework to support future research to improve PTP management in OB and OW children.

Classification of Computer-Aided Design-Computer-Aided Manufacturing Applications for the Reconstruction of Cranio-Maxillo-Facial Defects.

PURPOSE: To gain insight into the methodology of different computer-aided design-computer-aided manufacturing (CAD-CAM) applications for the reconstruction of cranio-maxillo-facial (CMF) defects. METHODS: We reviewed and analyzed the available literature pertaining to CAD-CAM for use in CMF reconstruction. RESULTS: We proposed a classification system of the techniques of implant and cutting, drilling, and/or guiding template design and manufacturing. The system consisted of 4 classes (I-IV). These classes combine techniques used for both the implant and template to most accurately describe the methodology used. CONCLUSIONS: Our classification system can be widely applied. It should facilitate communication and immediate understanding of the methodology of CAD-CAM applications for the reconstruction of CMF defects.

Modeling host genetic regulation of influenza pathogenesis in the collaborative cross.

Genetic variation contributes to host responses and outcomes following infection by influenza A virus or other viral infections. Yet narrow windows of disease symptoms and confounding environmental factors have made it difficult to identify polymorphic genes that contribute to differential disease outcomes in human populations. Therefore, to control for these confounding environmental variables in a system that models the levels of genetic diversity found in outbred populations such as humans, we used incipient lines of the highly genetically diverse Collaborative Cross (CC) recombinant inbred (RI) panel (the pre-CC population) to study how genetic variation impacts influenza associated disease across a genetically diverse population. A wide range of variation in influenza disease related phenotypes including virus replication, virus-induced inflammation, and weight loss was observed. Many of the disease associated phenotypes were correlated, with viral replication and virus-induced inflammation being predictors of virus-induced weight loss. Despite these correlations, pre-CC mice with unique and novel disease phenotype combinations were observed. We also identified sets of transcripts (modules) that were correlated with aspects of disease. In order to identify how host genetic polymorphisms contribute to the observed variation in disease, we conducted quantitative trait loci (QTL) mapping. We identified several QTL contributing to specific aspects of the host response including virus-induced weight loss, titer, pulmonary edema, neutrophil recruitment to the airways, and transcriptional expression. Existing whole-genome sequence data was applied to identify high priority candidate genes within QTL regions. A key host response QTL was located at the site of the known anti-influenza Mx1 gene. We sequenced the coding regions of Mx1 in the eight CC founder strains, and identified a novel Mx1 allele that showed reduced ability to inhibit viral replication, while maintaining protection from weight loss.

Expression quantitative trait Loci for extreme host response to influenza a in pre-collaborative cross mice.

Outbreaks of influenza occur on a yearly basis, causing a wide range of symptoms across the human population. Although evidence exists that the host response to influenza infection is influenced by genetic differences in the host, this has not been studied in a system with genetic diversity mirroring that of the human population. Here we used mice from 44 influenza-infected pre-Collaborative Cross lines determined to have extreme phenotypes with regard to the host response to influenza A virus infection. Global transcriptome profiling identified 2671 transcripts that were significantly differentially expressed between mice that showed a severe ("high") and mild ("low") response to infection. Expression quantitative trait loci mapping was performed on those transcripts that were differentially expressed because of differences in host response phenotype to identify putative regulatory regions potentially controlling their expression. Twenty-one significant expression quantitative trait loci were identified, which allowed direct examination of genes associated with regulation of host response to infection. To perform initial validation of our findings, quantitative polymerase chain reaction was performed in the infected founder strains, and we were able to confirm or partially confirm more than 70% of those tested. In addition, we explored putative causal and reactive (downstream) relationships between the significantly regulated genes and others in the high or low response groups using structural equation modeling. By using systems approaches and a genetically diverse population, we were able to develop a novel framework for identifying the underlying biological subnetworks under host genetic control during influenza virus infection.

Can a virtual patient trainer teach student nurses how to save lives--teaching nursing students about pediatric respiratory diseases.

INTRODUCTION: Virtual environments offer a variety of benefits and may be a powerful medium with which to provide nursing education. The objective of this study was to compare the achievement of learning outcomes of undergraduate nursing students when a virtual patient trainer or a traditional lecture was used to teach pediatric respiratory content. METHODS: This was a randomized, controlled, posttest design. A virtual pediatric hospital unit was populated with four virtual pediatric patients having different respiratory diseases that were designed to meet the same learning objectives as a traditional lecture. The study began in Spring 2010 with 93 Senior I, baccalaureate nursing students. Students were randomized to receive either a traditional lecture or an experience with a virtual patient trainer. Students' knowledge acquisition was evaluated using multiple-choice questions, and knowledge application was measured as timeliness of care in two simulated clinical scenarios using high-fidelity mannequins and standardized patients. RESULTS: Ninety-three students participated in the study, of which 46 were in the experimental group that received content using the virtual patient trainer. After the intervention, students in the experimental group had significantly higher knowledge acquisition (P = 0.004) and better knowledge application (P = 0.001) for each of the two scenarios than students in the control group. CONCLUSIONS: The purpose of this project was to compare a virtual patient trainer to a traditional lecture for the achievement of learning outcomes for pediatric respiratory content. Although the virtual patient trainer experience produced statistically better outcomes, the differences may not be clinically significant. The results suggest that a virtual patient trainer may be an effective substitute for the achievement of learning outcomes that are typically met using a traditional lecture format. Further research is needed to understand how best to integrate a virtual patient trainer into undergraduate nursing education.

Host regulatory network response to infection with highly pathogenic H5N1 avian influenza virus.

During the last decade, more than half of humans infected with highly pathogenic avian influenza (HPAI) H5N1 viruses have died, yet virus-induced host signaling has yet to be clearly elucidated. Airway epithelia are known to produce inflammatory mediators that contribute to HPAI H5N1-mediated pathogenicity, but a comprehensive analysis of the host response in this cell type is lacking. Here, we leveraged a system approach to identify and statistically validate signaling subnetworks that define the dynamic transcriptional response of human bronchial epithelial cells after infection with influenza A/Vietnam/1203/2004 (H5N1, VN1203). Importantly, we validated a subset of transcripts from one subnetwork in both Calu-3 cells and mice. A more detailed examination of two subnetworks involved in the immune response and keratinization processes revealed potential novel mediators of HPAI H5N1 pathogenesis and host response signaling. Finally, we show how these results compare to those for a less virulent strain of influenza virus. Using emergent network properties, we provide fresh insight into the host response to HPAI H5N1 virus infection and identify novel avenues for perturbation studies and potential therapeutic interventions for fatal HPAI H5N1 disease.

Genomic analysis reveals pre- and postchallenge differences in a rhesus macaque AIDS vaccine trial: insights into mechanisms of vaccine efficacy.

We have employed global transcriptional profiling of whole blood to identify biologically relevant changes in cellular gene expression in response to alternative AIDS vaccine strategies with subsequent viral challenge in a rhesus macaque vaccine model. Samples were taken at day 0 (prechallenge), day 14 (peak viremia), and week 12 (set point) from animals immunized with replicating adenovirus type 5 host range (Ad5hr) recombinant viruses expressing human immunodeficiency virus HIV(env)(89.6P), simian immunodeficiency virus SIV(gag)(239), or SIV(nef)(239) alone or in combination with two intramuscular boosts with HIV(89.6P)gp140ΔCFI protein (L. J. Patterson et al., Virology 374:322-337, 2008), and each treatment resulted in significant control of viremia following simian-human immunodeficiency virus SHIV(89.6P) challenge (six animals per group plus six controls). At day 0, 8 weeks after the last treatment, the microarray profiles revealed significant prechallenge differences between treatment groups; data from the best-protected animals led to identification of a network of genes related to B cell development and lymphocyte survival. At peak viremia, expression profiles of the immunized groups were extremely similar, and comparisons to control animals reflected immunological differences other than effector T cell functions. Suggested protective mechanisms for vaccinated animals included upregulation of interleukin-27, a cytokine known to inhibit lentivirus replication, and increased expression of complement components, which may synergize with vaccine-induced antibodies. Divergent expression profiles at set point for the immunized groups implied distinct immunological responses despite phenotypic similarities in viral load and CD4(+) T cell levels. Data for the gp140-boosted group provided evidence for antibody-dependent, cell-mediated viral control, whereas animals immunized with only the replicating Ad5hr recombinants exhibited a different evolution of the B cell compartment even at 3 months postchallenge. This study demonstrates the sensitivity and discrimination of gene expression profiling of whole blood as an analytical tool in AIDS vaccine trials, providing unique insights into in vivo mechanisms and potential correlates of protection.

Self-care strategies used by patients with lung cancer to promote quality of life.

PURPOSE/OBJECTIVES: To describe self-care strategies used by patients with lung cancer to promote quality of life (QOL). RESEARCH APPROACH: Qualitative study using a phenomenologic approach. SETTING: Cancer clinics in central Texas. PARTICIPANTS: Purposive sampling was used to enroll 10 adults with lung cancer who had completed primary treatment within the prior two years. METHODOLOGIC APPROACH: One-on-one, semistructured, audiotaped interviews were conducted. MAIN RESEARCH VARIABLES: QOL and self-care strategies. FINDINGS: Participants identified family and social support, functional independence, physical well-being, and spirituality as important aspects of QOL. Participants identified fatigue as the factor most negatively affecting QOL. Self-care strategies identified to improve QOL were primarily related to fatigue management. Rest was the primary self-care strategy reportedly recommended by healthcare providers, but this strategy was ineffective. Helpful self-care strategies included budgeting time and energy, maintaining contact with family and friends for support, and prayer. CONCLUSIONS: This study documents the negative effect of fatigue on QOL in patients with lung cancer. Use of rest to manage fatigue's pervasive negative effect on QOL is a common self-care strategy, reportedly recommended by healthcare providers, but is ineffective by itself to manage fatigue and improve QOL. INTERPRETATION: Healthcare providers should assess self-care strategies used by patients with lung cancer to promote improved QOL. Because fatigue has a documented negative effect on QOL in patients with lung cancer, providers should encourage the use of multidimensional strategies that have been supported by research evidence to manage fatigue and improve QOL.

Comparison of nitrile gloves and nitrile over Nomex gloves.

OBJECTIVES: Aeromedical flight crews must perform many tasks in flight requiring manual dexterity and fine precision. A common perception is that safety-enhancing fire-retardant gloves compromise patient care if worn during such tasks by providing added bulk and barrier to the hand. This study is a quantitative and qualitative analysis of this possible compromise to patient care. METHODS: Sixteen practicing flight nurses and respiratory therapists were asked to perform 10 different standard patient care tasks while wearing either nitrile gloves or a nitrile-Nomex glove pair. Tasks were timed, rated as completed successfully or not, and were subsequently judged subjectively by the participants. RESULTS: Whereas the time required to insert an intravenous catheter and to insert a central line while wearing only nitrile gloves was significantly faster than when wearing both gloves, the time to perform all other tasks was not significantly different. In subjective ratings, the nitrile glove alone was scored significantly better than the two-glove combination by the study participants. CONCLUSIONS: Comfort, dexterity, tactile discrimination, and ease of use were all adversely affected by wearing a Nomex glove under a nitrile glove. Although the differences in times for most tasks may not be clinically significant, the difference in the subjective parameters may be great enough to cause helicopter emergency medical services (HEMS) practitioners to not wear Nomex gloves under nitrile gloves while performing procedures.

Lethal influenza virus infection in macaques is associated with early dysregulation of inflammatory related genes.

The enormous toll on human life during the 1918-1919 Spanish influenza pandemic is a constant reminder of the potential lethality of influenza viruses. With the declaration by the World Health Organization of a new H1N1 influenza virus pandemic, and with continued human cases of highly pathogenic H5N1 avian influenza virus infection, a better understanding of the host response to highly pathogenic influenza viruses is essential. To this end, we compared pathology and global gene expression profiles in bronchial tissue from macaques infected with either the reconstructed 1918 pandemic virus or the highly pathogenic avian H5N1 virus A/Vietnam/1203/04. Severe pathology was observed in respiratory tissues from 1918 virus-infected animals as early as 12 hours after infection, and pathology steadily increased at later time points. Although tissues from animals infected with A/Vietnam/1203/04 also showed clear signs of pathology early on, less pathology was observed at later time points, and there was evidence of tissue repair. Global transcriptional profiles revealed that specific groups of genes associated with inflammation and cell death were up-regulated in bronchial tissues from animals infected with the 1918 virus but down-regulated in animals infected with A/Vietnam/1203/04. Importantly, the 1918 virus up-regulated key components of the inflammasome, NLRP3 and IL-1beta, whereas these genes were down-regulated by A/Vietnam/1203/04 early after infection. TUNEL assays revealed that both viruses elicited an apoptotic response in lungs and bronchi, although the response occurred earlier during 1918 virus infection. Our findings suggest that the severity of disease in 1918 virus-infected macaques is a consequence of the early up-regulation of cell death and inflammatory related genes, in which additive or synergistic effects likely dictate the severity of tissue damage.

Computational identification of hepatitis C virus associated microRNA-mRNA regulatory modules in human livers.

BACKGROUND: Hepatitis C virus (HCV) is a major cause of chronic liver disease by infecting over 170 million people worldwide. Recent studies have shown that microRNAs (miRNAs), a class of small non-coding regulatory RNAs, are involved in the regulation of HCV infection, but their functions have not been systematically studied. We propose an integrative strategy for identifying the miRNA-mRNA regulatory modules that are associated with HCV infection. This strategy combines paired expression profiles of miRNAs and mRNAs and computational target predictions. A miRNA-mRNA regulatory module consists of a set of miRNAs and their targets, in which the miRNAs are predicted to coordinately regulate the level of the target mRNA. RESULTS: We simultaneously profiled the expression of cellular miRNAs and mRNAs across 30 HCV positive or negative human liver biopsy samples using microarray technology. We constructed a miRNA-mRNA regulatory network, and using a graph theoretical approach, identified 38 miRNA-mRNA regulatory modules in the network that were associated with HCV infection. We evaluated the direct miRNA regulation of the mRNA levels of targets in regulatory modules using previously published miRNA transfection data. We analyzed the functional roles of individual modules at the systems level by integrating a large-scale protein interaction network. We found that various biological processes, including some HCV infection related canonical pathways, were regulated at the miRNA level during HCV infection. CONCLUSION: Our regulatory modules provide a framework for future experimental analyses. This report demonstrates the utility of our approach to obtain new insights into post-transcriptional gene regulation at the miRNA level in complex human diseases.

Early and sustained innate immune response defines pathology and death in nonhuman primates infected by highly pathogenic influenza virus.

The mechanisms responsible for the virulence of the highly pathogenic avian influenza (HPAI) and of the 1918 pandemic influenza virus in humans remain poorly understood. To identify crucial components of the early host response during these infections by using both conventional and functional genomics tools, we studied 34 cynomolgus macaques (Macaca fascicularis) to compare a 2004 human H5N1 Vietnam isolate with 2 reassortant viruses possessing the 1918 hemagglutinin (HA) and neuraminidase (NA) surface proteins, known conveyors of virulence. One of the reassortants also contained the 1918 nonstructural (NS1) protein, an inhibitor of the host interferon response. Among these viruses, HPAI H5N1 was the most virulent. Within 24 h, the H5N1 virus produced severe bronchiolar and alveolar lesions. Notably, the H5N1 virus targeted type II pneumocytes throughout the 7-day infection, and induced the most dramatic and sustained expression of type I interferons and inflammatory and innate immune genes, as measured by genomic and protein assays. The H5N1 infection also resulted in prolonged margination of circulating T lymphocytes and notable apoptosis of activated dendritic cells in the lungs and draining lymph nodes early during infection. While both 1918 reassortant viruses also were highly pathogenic, the H5N1 virus was exceptional for the extent of tissue damage, cytokinemia, and interference with immune regulatory mechanisms, which may help explain the extreme virulence of HPAI viruses in humans.

Aberrant beta-catenin signaling in tuberous sclerosis.

The pathology associated with tuberous sclerosis complex (TSC) shows diverse phenotypes that suggest abnormal signaling of multiple pathways. Besides the negative regulatory role of the TSC1/TSC2 proteins on mTOR, we have reported an effect on beta-catenin signaling at the level of the degradation complex in vitro. The TSC1/TSC2 complex associates with GSK3 and Axin and promotes beta-catenin degradation to inhibit Wnt-stimulated TCF/LEF-dependent transcription. Here, we show that beta-catenin and its effectors, cyclin D1 and connexin 43, were up-regulated in TSC-related angiomyolipomas and lymphangioleiomyomatosis. This was supported by the failure of three disease-causing TSC2 missense mutants to inhibit Wnt signaling. Further, the interaction between TSC1/TSC2 and components of the beta-catenin degradation complex was dependent on Wnt stimulation such that binding of tuberin to GSK3 and Axin was reduced in the presence of Wnt whereas the tuberin-Dishevelled interaction was increased. GSK3 activity played a role in regulating the assembly/stability of the degradation complex. Inhibition of GSK3 by lithium chloride reduced its association with TSC1 whereas disruption of GSK3-phosphorylation sites in TSC1 reduced interaction between TSC2 and TSC1. Collectively, our data provide further evidence that beta-catenin signaling plays a role in TSC pathogenesis in vivo and suggest a novel role of GSK3 in modulating the TSC1/TSC2 complex through TSC1 phosphorylation.

The effect of seated exercise on fatigue and quality of life in women with advanced breast cancer.

PURPOSE/OBJECTIVES: To examine the effects of a seated exercise program on fatigue and quality of life (QOL) in women with metastatic breast cancer. DESIGN: Randomized, controlled, longitudinal trial. SETTING: Outpatient clinic of a comprehensive cancer center. SAMPLE: Convenience sample of 38 women who were beginning outpatient chemotherapy. METHODS: Subjects were randomized to a control or intervention group; the intervention was performance of a seated exercise program using home videotape three times per week for four cycles of chemotherapy. All subjects completed the Functional Assessment of Chronic Illness Therapy Fatigue Version IV (FACIT F) at baseline and at the time of the next three cycles. Subjects were asked to document the frequency, duration, and intensity of all exercise participation on monthly calendars. MAIN RESEARCH VARIABLES: Exercise, fatigue, and QOL. FINDINGS: 32 subjects, 16 per group, completed the study follow-up. With a mixed modeling approach, total FACIT F scores for the entire sample declined at a significant rate (p = 0.003) beginning with cycle 3 but at a slower rate for the experimental group (p = 0.02). Fatigue scores indicated less increase and physical well-being subscale scores showed less decline for the experimental group (p = 0.008 and p = 0.02, respectively). CONCLUSIONS: Women with advanced breast cancer randomized to the seated exercise intervention had a slower decline in total and physical well-being and less increase in fatigue scores starting with the third cycle of chemotherapy. IMPLICATIONS FOR NURSING: Seated exercise may be a feasible exercise program for women with advanced cancer for controlling fatigue and improving physical well-being.

Improved detection of cystic fibrosis mutations in infertility patients with DNA sequence analysis.

BACKGROUND: Accurate determination of mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene is critical for genetic counselling and treatment of obstructive azoospermia. Of concern is that detection rates with routine CFTR mutation panels vary widely depending on patient ancestry; and such panels have limited value for azoospermic patients, who are more likely to carry rare mutations. An alternative approach offers comprehensive, CFTR mutation analysis by a DNA sequence method. We investigated whether this method could improve CFTR detection rates in men with obstructive azoospermia in a prospective study of men with obstructive azoospermia and their partners who were referred for genetic counselling and testing at one of two institutions. METHODS: Sixteen patients with congenital absence of the vas deferens (CAVD, n = 14) or idiopathic obstructive azoospermia (n = 2) were studied. DNA from all patients was analysed for mutations by the DNA sequence method. In addition to this method, six men underwent CFTR analysis by a common 25 or 31 mutation panel coupled with poly T analysis. In 10 subjects, common mutation panel findings were inferred from DNA sequence method results. RESULTS: Overall, 12/16 (75%) azoospermic patients had one or more CFTR mutations and/or 5T alleles, including 12 mutations in 10 patients (two compound heterozygotes) and seven 5T alleles in six patients (one homozygote). The sequence method detected all mutations and three variants of unknown significance. By comparison, the common mutation panels detected only 3/12 mutations (25%) and 0/3 variants. CONCLUSION: The DNA sequence method detects more CFTR mutations than common mutation panels. Given the serious, clinical consequences of transmitting such mutations, this study underscores the importance of accurate, CFTR mutation detection in men with obstructive azoospermia and their partners.