Mutation profiles in early-stage lung squamous cell carcinoma with clinical follow-up and correlation with markers of immune function.

Background: Lung squamous cell carcinoma (LUSC) accounts for 20­30% of non-small cell lung cancers (NSCLCs). There are limited treatment strategies for LUSC in part due to our inadequate understanding of the molecular underpinnings of the disease. We performed whole-exome sequencing (WES) and comprehensive immune profiling of a unique set of clinically annotated early-stage LUSCs to increase our understanding of the pathobiology of this malignancy. Methods: Matched pairs of surgically resected stage I-III LUSCs and normal lung tissues (n = 108) were analyzed by WES. Immunohistochemistry and image analysis-based profiling of 10 immune markers were done on a subset of LUSCs (n = 91). Associations among mutations, immune markers and clinicopathological variables were statistically examined using analysis of variance and Fisher's exact test. Cox proportional hazards regression models were used for statistical analysis of clinical outcome. Results: This early-stage LUSC cohort displayed an average of 209 exonic mutations per tumor. Fourteen genes exhibited significant enrichment for somatic mutation: TP53, MLL2, PIK3CA, NFE2L2, CDH8, KEAP1, PTEN, ADCY8, PTPRT, CALCR, GRM8, FBXW7, RB1 and CDKN2A. Among mutated genes associated with poor recurrence-free survival, MLL2 mutations predicted poor prognosis in both TP53 mutant and wild-type LUSCs. We also found that in treated patients, FBXW7 and KEAP1 mutations were associated with poor response to adjuvant therapy, particularly in TP53-mutant tumors. Analysis of mutations with immune markers revealed that ADCY8 and PIK3CA mutations were associated with markedly decreased tumoral PD-L1 expression, LUSCs with PIK3CA mutations exhibited elevated CD45ro levels and CDKN2A-mutant tumors displayed an up-regulated immune response. Conclusion(s): Our findings pinpoint mutated genes that may impact clinical outcome as well as personalized strategies for targeted immunotherapies in early-stage LUSC.

Whole-exome sequencing and immune profiling of early-stage lung adenocarcinoma with fully annotated clinical follow-up.

Background: Lung adenocarcinomas (LUADs) lead to the majority of deaths attributable to lung cancer. We performed whole-exome sequencing (WES) and immune profiling analyses of a unique set of clinically annotated early-stage LUADs to better understand the pathogenesis of this disease and identify clinically relevant molecular markers. Methods: We performed WES of 108 paired stage I-III LUADs and normal lung tissues using the Illumina HiSeq 2000 platform. Ten immune markers (PD-L1, PD-1, CD3, CD4, CD8, CD45ro, CD57, CD68, FOXP3 and Granzyme B) were profiled by imaging-based immunohistochemistry (IHC) in a subset of LUADs (n = 92). Associations among mutations, immune markers and clinicopathological variables were analyzed using ANOVA and Fisher's exact test. Cox proportional hazards regression models were used for multivariate analysis of clinical outcome. Results: LUADs in this cohort exhibited an average of 243 coding mutations. We identified 28 genes with significant enrichment for mutation. SETD2-mutated LUADs exhibited relatively poor recurrence- free survival (RFS) and mutations in STK11 and ATM were associated with poor RFS among KRAS-mutant tumors. EGFR, KEAP1 and PIK3CA mutations were predictive of poor response to adjuvant therapy. Immune marker analysis revealed that LUADs in smokers and with relatively high mutation burdens exhibited increased levels of immune markers. Analysis of immunophenotypes revealed that LUADs with STK11 mutations exhibited relatively low levels of infiltrating CD4+/CD8+ T-cells indicative of a muted immune response. Tumoral PD-L1 was significantly elevated in TP53 mutant LUADs whereas PIK3CA mutant LUADs exhibited markedly down-regulated PD-L1 expression. LUADs with TP53 or KEAP1 mutations displayed relatively increased CD57 and Granzyme B levels indicative of augmented natural killer (NK) cell infiltration. Conclusion(s): Our study highlights molecular and immune phenotypes that warrant further analysis for their roles in clinical outcomes and personalized immune-based therapy of LUAD.

Evaluation of take-home exposure and risk associated with the handling of clothing contaminated with chrysotile asbestos.

The potential for para-occupational (or take-home) exposures from contaminated clothing has been recognized for the past 60 years. To better characterize the take-home asbestos exposure pathway, a study was performed to measure the relationship between airborne chrysotile concentrations in the workplace, the contamination of work clothing, and take-home exposures and risks. The study included air sampling during two activities: (1) contamination of work clothing by airborne chrysotile (i.e., loading the clothing), and (2) handling and shaking out of the clothes. The clothes were contaminated at three different target airborne chrysotile concentrations (0-0.1 fibers per cubic centimeter [f/cc], 1-2 f/cc, and 2-4 f/cc; two events each for 31-43 minutes; six events total). Arithmetic mean concentrations for the three target loading levels were 0.01 f/cc, 1.65 f/cc, and 2.84 f/cc (National Institute of Occupational Health and Safety [NIOSH] 7402). Following the loading events, six matched 30-minute clothes-handling and shake-out events were conducted, each including 15 minutes of active handling (15-minute means; 0.014-0.097 f/cc) and 15 additional minutes of no handling (30-minute means; 0.006-0.063 f/cc). Percentages of personal clothes-handling TWAs relative to clothes-loading TWAs were calculated for event pairs to characterize exposure potential during daily versus weekly clothes-handling activity. Airborne concentrations for the clothes handler were 0.2-1.4% (eight-hour TWA or daily ratio) and 0.03-0.27% (40-hour TWA or weekly ratio) of loading TWAs. Cumulative chrysotile doses for clothes handling at airborne concentrations tested were estimated to be consistent with lifetime cumulative chrysotile doses associated with ambient air exposure (range for take-home or ambient doses: 0.00044-0.105 f/cc year).

An evidence-based analysis of epidemiologic associations between lymphatic and hematopoietic cancers and occupational exposure to gasoline.

The presence of benzene in motor gasoline has been a health concern for potential increased risk of acute myelogenous leukemia and perhaps other lymphatic/hematopoietic cancers for approximately 40 years. Because of the widespread and increasing use of gasoline by consumers and the high exposure potential of occupational cohorts, a thorough understanding of this issue is important. The current study utilizes an evidence-based approach to examine whether or not the available epidemiologic studies demonstrate a strong and consistent association between occupational exposure to gasoline and lymphatic/hematopoietic cancers. Among 67 epidemiologic studies initially identified, 54 were ranked according to specific criteria relating to the relevance and robustness of each study for answering the research question. The 30 highest-ranked studies were sorted into three tiers of evidence and were analyzed for strength, specificity, consistency, temporality, dose-response trends and coherence. Meta statistics were also calculated for each general and specific lymphatic/hematopoietic cancer category with adequate data. The evidence-based analysis did not confirm any strong and consistent association between occupational exposure to gasoline and lymphatic/hematopoietic cancers based on the epidemiologic studies available to date. These epidemiologic findings, combined with the evidence showing relatively low occupational benzene vapor exposures associated with gasoline formulations during the last three decades, suggest that current motor gasoline formulations are not associated with increased lymphatic/hematopoietic cancer risks related to benzene.

Characterization of formaldehyde exposure resulting from the use of four professional hair straightening products.

An exposure simulation study was conducted to characterize potential formaldehyde exposures of salon workers and clients during keratin hair smoothing treatments. Four different hair treatment brands (Brazilian Blowout, Coppola, Global Keratin, and La Brasiliana) were applied to separate human hair wigs mounted on mannequin heads. Short-term (6-16 min) and long-term (41-371 min) personal and area samples (at distances of 0.5 to 3.0 m from the source) were collected during each treatment for the 1-day simulation. A total of 88 personal, area, and clearance samples were collected. Results were analyzed based on task sampling (blow-dry, flat-iron), treatment sampling (per hair product), and time-weighted averages (per hair treatment, four consecutive treatments). Real-time monitoring of tracer gas levels, for determining the air exchange rate, and formaldehyde levels were logged throughout the simulation. Bulk samples of each hair treatment were collected to identify and quantify formaldehyde and other chemical components that may degrade to formaldehyde under excessive heat. Mean airborne concentrations of formaldehyde ranged from 0.08-3.47 ppm during blow-dry and 0.08-1.05 ppm during flat-iron. During each treatment, the mean airborne concentrations ranged from 0.02-1.19 ppm throughout different zones of the salon. Estimated 8-hr time-weighted averages for one treatment per day ranged from 0.02 ppm for La Brasiliana to 0.08-0.16 ppm for Brazilian Blowout. For four treatments per day, means ranged from 0.04-0.05 ppm for La Brasiliana to 0.44-0.75 ppm for Brazilian Blowout. Using all four products in one day resulted in estimated 8-hr time-weighted averages ranging from 0.17-0.29 ppm. Results from bulk sampling reported formaldehyde concentrations of 11.5% in Brazilian Blowout, 8.3% in Global Keratin, 3% in Coppola, and 0% in La Brasiliana. Other products that degrade into formaldehyde were detected in Global Keratin, Coppola, and La Brasiliana. The results of this study show that professional hair smoothing treatments--even those labeled "formaldehyde-free"--have the potential to produce formaldehyde concentrations that meet or exceed current occupational exposure limits.

Airborne asbestos concentrations associated with heavy equipment brake removal.

Asbestos-containing brake linings were used in heavy-duty construction equipment such as tractors, backhoes, and bulldozers prior to the 1980s. While several published studies have evaluated exposures to mechanics during brake repair work, most have focused on automobiles and light trucks, not on heavy agricultural or construction vehicles. The purpose of this study is to characterize the airborne concentration of asbestos to workers and bystanders from brake wear debris during brake removal from 12 loader/backhoes and tractors manufactured between 1960 and 1980. Asbestos content in brake lining (average 20% chrysotile by polarized light microscopy) and brake wear debris [average 0.49% chrysotile by transmission electron microscopy (TEM)] was also quantified. Breathing zone samples on the lapel of mechanics (n = 44) and area samples at bystander (n = 34), remote (n = 22), and ambient (n = 12) locations were collected during 12 brake changes and analyzed using phase contrast microscopy (PCM) [National Institute for Occupational Safety and Health (NIOSH) 7400] and TEM (NIOSH 7402). In addition, the fiber distribution by size and morphology was evaluated according to the International Organization for Standardization method for asbestos. Applying the ratio of asbestos fibers:total fibers (including non-asbestos) as determined by TEM to the PCM results, the average airborne chrysotile concentrations (PCM equivalent) were 0.024 f/cc for the mechanic and 0.009 f/cc for persons standing 1.2-3.1 m from the activity during the period of exposure ( approximately 0.5 to 1 h). Considering the time involved in the activity, and assuming three brake jobs per shift, these results would convert to an average 8-h time-weighted average of 0.009 f/cc for a mechanic and 0.006 f/cc for a bystander. The results indicate that (i) the airborne concentrations for worker and bystander samples were significantly less than the current occupational exposure limit of 0.1 f/cc; (ii) approximately 2% of respirable fibers were >20 microm in length; and (iii) approximately 95% of chrysotile in the brake linings degraded in the friction process. The industrial hygiene data presented here should be useful for conducting retrospective and current exposure assessments of individuals, as well as hazard assessments of work activities that involve repairing and replacing asbestos-containing brakes in heavy construction equipment.

Historical analysis of airborne beryllium concentrations at a copper beryllium machining facility (1964-2000).

Copper beryllium alloys are the most commonly used form of beryllium; however, there have been few studies assessing occupational exposure in facilities that worked exclusively with this alloy versus those where pure metal or beryllium oxide may also have been present. In this paper, we evaluated the airborne beryllium concentrations at a machining plant using historical industrial hygiene samples collected between 1964 and 2000. With the exception of a few projects conducted in the 1960s, it is believed that >95% of the operations used copper beryllium alloy exclusively. Long-term (>120 min) and short-term (<120 min) personal and area samples were collected during a variety of activities including machining of copper beryllium-containing parts, as well as finishing operations (e.g., deburring and polishing) and decontamination of machinery. A total of 580 beryllium air samples were analyzed (311 personal and 269 area samples). The average concentration based on area samples (1964-2000) was 0.021 microg m(-3) (SD 0.17 microg m(-3); range 0.00012-2.5 microg m(-3)); 68.8% were below the analytical limit of detection (LOD). The average airborne beryllium concentration, based on all personal samples available from 1964 through the end of 2000 (n = 311), was 0.026 microg m(-3) (SD 0.059 microg m(-3); range 0.019-0.8 microg m(-3)); 97.4% were below the LOD. Personal samples collected from machinists (n = 78) had an average airborne concentration of 0.021 microg m(-3) (SD 0.014 microg m(-3); range 0.019-0.14 microg m(-3)); 97.4% were below the LOD. Airborne concentrations were consistently below the Occupational Safety and Health Administration permissible exposure limit for beryllium (2 microg m(-3)). Overall, the data indicate that for machining operations involving copper beryllium, the airborne concentrations for >95% of the samples were below the contemporaneous occupational exposure limits or the 1999 Department of Energy action level of 0.2 microg m(-3) and, in most cases, were below the LOD.

Non-small-cell lung cancer dimensions: CT-pathological correlation and interobserver variation.

The aim of the study was to identify the most accurate CT window level setting for the measurement of non-small-cell lung cancer to optimise CT planning for radiotherapy treatment. 27 patients who underwent resection for non-small-cell lung cancer in a single institution were studied. The maximal superior-inferior, anteroposterior and mediolateral dimensions of the resected tumours were measured by a consultant pathologist. Two radiologists made corresponding measurements using pre-operative CT scans independently of each other and of the pathologist's findings. The measurements were obtained using four different CT window settings. The mean pathological size of the superior-inferior tumours, the anteroposterior tumours and the mediolateral tumours was 32 mm, 28 mm and 25 mm, respectively. A total of 648 CT measurements were taken, of which 321 were within +/-5 mm of the pathological size (49.5%). There was significant interobserver variability between the two radiologists. There was poor correlation between the pathological and radiological measurements of tumour size. Significant interobserver variability was noted between the two radiologists and no window setting could be identified as being superior in accurately assessing the tumour size.

Motor development and gender effects on stretch-shortening cycle performance.

A substantial part of human movement such as jumping, hopping, leaping and other bounding movements are improved by making a counter-movement. These activities are often described as stretch shortening cycle (SSC) movements. The aim of this study was to determine whether the SSC affects performance in vertical jump in children to the same extent as it does in adults. Comparisons were made between counter-movement jump (CMJ) and squat jump (SJ) performance in children and adults. The ratio of take-off velocity between jumps was used to measure performance of the SSC. Two groups of subjects comprising of 22 adults and 20 children performed three CMJ's and three SJ's from a force platform. Impulse, take-off velocity and power were obtained by numerical integration of the force n time traces. Performance was calculated from the velocity at take off. Both groups jumped significantly higher in the CMJ but there was a higher degree of variability in the performance of the children. The results indicated that children could utilise a SSC to enhance jumping performance. Variability in the take-off velocities in children, particularly in the SJ suggests the children performed this jump non-optimally.

Expression, purification, and bioassay of human stanniocalcin from baculovirus-infected insect cells and recombinant CHO cells.

Stanniocalcin is a calcium- and phosphate-regulating glycoprotein hormone that was first described in fish where it functions in preventing hypercalcemia. Human cDNA clones encoding the homolog of stanniocalcin have been recently isolated. In this study, the full-length cDNA coding for human stanniocalcin (hSTC) was cloned into both baculovirus and CHO expression vectors. Recombinant hSTC was then produced efficiently from both baculovirus-infected insect cells and CHO cells in large-scale bioreactors. Purification protocols were developed and used to purify recombinant hSTC from both sources in four chromatography steps. The hSTCs from both expression systems were secreted as glycosylated proteins and as disulfide-linked homodimers. The results from glycosylation studies indicated that stanniocalcin from both sources contained N-linked oligosaccharides but no O-linked sugars. In an in vivo bioassay based on the inhibition of gill calcium transport in fishes, the baculovirus and CHO-expressed protein showed biological activity which is dose dependent. The inhibitory effects of hSTC produced from both systems were essentially equipotent in fishes, despite the differences in glycosylation. Consequently, the precise role of the carbohydrate moiety in recombinant hSTC remains to be determined.

Seizure disorders and pregnancy.

Care of a pregnant woman with seizure disorder is a challenge for the maternal-child nurse. Epilepsy affects 0.5% of all pregnant women. These women face possible risks of increased seizure frequency, reduced anti-epileptic drug levels, and, although rare, life-threatening status epilepticus. Their neonates are at risk for congenital malformations and possible bleeding disorders. Comprehensive nursing care based on a sound understanding of medical, pharmacologic, and nursing principles is crucial for optimal maternal/neonatal outcomes. Specific nursing implications and interventions during the preconceptual, prenatal, intrapartal, and postpartal periods are presented.

Relationship between cyclic variation of fetal heart rate patterns and cord pH in preterm gestations.

The presence or absence of cyclic variation (cyclicity) of fetal heart rate patterns was prospectively investigated in fetuses between 25 and 32 weeks' gestation. All fetuses were delivered by cesarean section before the onset of labor. The relationship between the presence or absence of fetal heart rate cyclicity and fetal acidosis, as determined by cord pH measurements, was investigated. The sensitivity, specificity, positive predictive value, and negative predictive value of this method in predicting fetal acidosis were: 100, 90, 50, and 100%, respectively. All fetuses with reactive nonstress tests also had fetal heart rate cyclicity present and none were acidotic at birth. The majority of fetuses (68.8%) with nonreactive nonstress tests had fetal heart rate cyclicity present and none were acidotic; fetuses with nonreactive nonstress tests and absence of fetal heart rate cyclicity were acidotic in 50% of the cases. These preliminary data suggest that the presence or absence of fetal heart rate cyclicity may help to select the healthy preterm fetuses with nonreactive nonstress testing who are in good condition and therefore in no need for further testing.

The relationships among the fetal biophysical profile, umbilical cord pH, and Apgar scores.

The relationships among the fetal biophysical profile, umbilical cord blood pH, and Apgar scores were investigated in 124 patients undergoing cesarean section before the onset of labor. Based on a standard for the diagnosis of fetal acidosis of an umbilical cord arterial pH less than 7.20, the sensitivity, specificity, and positive and negative predictive values of the fetal biophysical profile score were 90%, 96%, 82%, and 98%, respectively. When the combination of a nonreactive nonstress test and absent fetal breathing was used as the "abnormal test," the sensitivity, specificity, and positive and negative predictive values were 100%, 92%, 71%, and 100%, respectively. The efficacy of the fetal biophysical profile to indicate fetal acidosis was found to be superior to the 1- and 5-minute Apgar scores in sensitivity and positive predictive value. These data suggest that the biophysical profile is very accurate in the identification of the fetus with acidemia. The first manifestations of fetal acidosis are nonreactive nonstress testing and loss of fetal breathing; in advanced acidemia fetal movements and fetal tone are compromised. A new protocol of antepartum fetal evaluation is suggested based on individual biophysical components rather than the score alone.

The relationship between fetal biophysical profile and cord pH in patients undergoing cesarean section before the onset of labor.

In a prospective study of 124 patients undergoing cesarean section before the onset of labor, the fetal biophysical profile was found to have a significant relationship with umbilical cord blood pH (artery and vein). Using cord arterial pH less than 7.20 as a standard for the diagnosis of fetal acidosis, the sensitivity, specificity, and positive and negative predictive values of the fetal biophysical profile score were 90, 96, 82, and 98%, respectively. When the combination of nonreactive nonstress test and absent fetal breathing was used as the "abnormal test," the sensitivity, specificity, and positive and negative predictive values were 100, 92, 71, and 100%, respectively. The first manifestations of fetal acidosis are nonreactive nonstress testing and loss of fetal breathing; in advanced acidemia, fetal movements and fetal tone are compromised. A protocol of antepartum fetal evaluation is suggested based upon the individual biophysical components rather than the score alone.

Group B streptococcus: the pregnant woman and her neonate.

Within the past 10 years, group B streptococcal disease has emerged as a major causative agent in neonatal morbidity and mortality. A brief history of this pathogen and its epidemiologic factors regarding maternal colonization and carriage is presented. Group B streptococcal disease in the neonate and concurrent treatment and prevention modalities also are presented. A case study is included. Suggestions for nursing assessment, intervention, and future considerations are offered.