Pharmacologic Tumor PDL1 Depletion with Cefepime or Ceftazidime Promotes DNA Damage and Sensitivity to DNA-Damaging Agents.

The interaction between tumor surface-expressed PDL1 and immune cell PD1 for the evasion of antitumor immunity is well established and is targeted by FDA-approved anti-PDL1 and anti-PD1 antibodies. Nonetheless, recent studies highlight the immunopathogenicity of tumor-intrinsic PDL1 signals that can contribute to the resistance to targeted small molecules, cytotoxic chemotherapy, and αPD1 immunotherapy. As genetic PDL1 depletion is not currently clinically tractable, we screened FDA-approved drugs to identify those that significantly deplete tumor PDL1. Among the candidates, we identified the ß-lactam cephalosporin antibiotic cefepime as a tumor PDL1-depleting drug (PDD) that increases tumor DNA damage and sensitivity to DNA-damaging agents in vitro in distinct aggressive mouse and human cancer lines, including glioblastoma multiforme, ovarian cancer, bladder cancer, and melanoma. Cefepime reduced tumor PDL1 post-translationally through ubiquitination, improved DNA-damaging-agent treatment efficacy in vivo in immune-deficient and -proficient mice, activated immunogenic tumor STING signals, and phenocopied specific genetic PDL1 depletion effects. The ß-lactam ring and its antibiotic properties did not appear contributory to PDL1 depletion or to these treatment effects, and the related cephalosporin ceftazidime produced similar effects. Our findings highlight the rapidly translated potential for PDDs to inhibit tumor-intrinsic PDL1 signals and improve DNA-damaging agents and immunotherapy efficacy.

Cutaneous Manifestation as Initial Presentation of Metastatic Breast Cancer: A Systematic Review.

Metastatic breast cancer initially may present with cutaneous lesions. The goal of this systematic review was to evaluate available reports where the initial discovery of primary breast cancer occurred through the diagnosis of metastatic cutaneous lesions. We aimed to better understand these cases and the role of dermatologists in their diagnosis. A review of the literature for case reports and retrospective studies was conducted using the following databases: MEDLINE/PubMed, EMBASE, Cochrane library, CINAHL, and EBSCO. The PRISMA guidelines were utilized. Studies were included if they reported a cutaneous metastasis of a primary breast cancer in females. Studies were excluded if skin metastasis occurred in a patient with a history of breast cancer. Thirty-six publications were identified. Among these, 27 were case reports, and 9 were retrospective reviews. An enhanced understanding of how these cutaneous metastases present may be of clinical benefit to physicians, particularly dermatologists.

Neuronal nitric oxide synthase mediates insulin- and oxidative stress-induced glucose uptake in skeletal muscle myotubes.

Previously published studies strongly suggested that insulin- and exercise-induced skeletal muscle glucose uptake require nitric oxide (NO) production. However, the signal transduction mechanisms by which insulin and contraction regulated NO production and subsequent glucose transport are not known. In the present study, we utilized the myotube cell lines treated with insulin or hydrogen peroxide, the latter to mimic contraction-induced oxidative stress, to characterize these mechanisms. We found that insulin stimulation of neuronal nitric oxide synthase (nNOS) phosphorylation, NO production, and GLUT4 translocation were all significantly reduced by inhibition of either nNOS or Akt2. Hydrogen peroxide (H2O2) induced phosphorylation of nNOS at the same residue as did insulin, and also stimulated NO production and GLUT4 translocation. nNOS inhibition prevented H2O2-induced GLUT4 translocation. AMP activated protein kinase (AMPK) inhibition prevented H2O2 activation and phosphorylation of nNOS, leading to reduced NO production and significantly attenuated GLUT4 translocation. We conclude that nNOS phosphorylation and subsequently increased NO production are required for both insulin- and H2O2-stimulated glucose transport. Although the two stimuli result in phosphorylation of the same residue on nNOS, they do so through distinct protein kinases. Thus, insulin and H2O2-activated signaling pathways converge on nNOS, which is a common mediator of glucose uptake in both pathways. However, the fact that different kinases are utilized provides a basis for the use of exercise to activate glucose transport in the face of insulin resistance.

Effect of Obesity on Perioperative Outcomes After Robotic-Assisted Pulmonary Lobectomy.

OBJECTIVE: We investigated whether higher body mass index (BMI) affects perioperative and postoperative outcomes after robotic-assisted video-thoracoscopic pulmonary lobectomy. METHODS: We retrospectively studied all patients who underwent robotic-assisted pulmonary lobectomy by one surgeon between September 2010 and January 2015. Patients were grouped according to the World Health Organization's definition of obesity, with "obese" being defined as BMI >30.0 kg/m2. Perioperative outcomes, including intraoperative estimated blood loss (EBL) and postoperative complication rates, were compared. RESULTS: Over 53 months, 287 patients underwent robotic-assisted pulmonary lobectomy, with 7 patients categorized as "underweight," 94 patients categorized as "normal weight," 106 patients categorized as "overweight," and 80 patients categorized as "obese." Because of the relatively low sample size, "underweight" patients were excluded from this study, leaving a total cohort of 280 patients. There was no significant difference in intraoperative complication rates, conversion rates, perioperative outcomes, or postoperative complication rates among the 3 groups, except for lower risk of prolonged air leaks ≥7 days and higher risk of pneumonia in patients with obesity. CONCLUSIONS: Patients with obesity do not have increased risk of intraoperative or postoperative complications, except for pneumonia, compared with "normal weight" and "overweight" patients. Robotic-assisted pulmonary lobectomy is safe and effective for patients with high BMI.

Effect of advanced age on peri-operative outcomes after robotic-assisted pulmonary lobectomy: Retrospective analysis of 287 consecutive cases.

OBJECTIVES: We investigated whether advanced age affects peri-operative outcomes after robotic-assisted pulmonary lobectomies. MATERIALS AND METHODS: We retrospectively analyzed patients who underwent robotic-assisted lobectomy by one surgeon over a 5-year period. Rates of postoperative complications were compared according to age group. Other outcomes, such as intraoperative complications, hospital length of stay (LOS), and in-hospital mortality, were also compared. RESULTS: A total of 287 patients were included (mean age 67.1yr). Group A had 65 patients of advanced age≥75yr (range 75-87yr; 37 men, 28 women); Group B had 222 patients aged <75yr (range 29-74yr; 95 men, 127 women). Group A had 10/65 (15.4%) patients with robotic-related intraoperative complications, compared to 10/222 (4.5%) for Group B (p=0.002), with the most frequent intraoperative complications being bleeding from a pulmonary vessel (10.8% vs. 4.5%, p=0.06), bronchial injury (3.1% vs. 0.9%, p=0.18), and injury to the phrenic or recurrent laryngeal nerve (1.5% vs. 0.4%, p=0.33). There were 28/65 (43.1%) patients in Group A with postoperative complications compared to 76/222 (34.2%) in Group B (p=0.19). While operative times were similar (p=0.42), Group A had longer median hospital LOS of 6±0.9days compared to 4±0.3days for Group B (p=0.02). CONCLUSION: While younger patients have lower risk of robotic-related intraoperative complications and shorter hospital LOS, elderly patients do not have increased overall or emergent conversion rates to open lobectomy, overall postoperative complications rates, or in-house mortality compared to younger patients. Thus, robotic-assisted pulmonary lobectomy is feasible and relatively safe for patients of advanced age.

Efficacy of lymph node dissection during robotic-assisted lobectomy for non-small cell lung cancer: retrospective review of 159 consecutive cases.

BACKGROUND: We investigated whether robotic-assisted surgery improves mediastinal lymph node dissection (MLND). METHODS: We analyzed patients (pts) who underwent robotic-assisted video-assisted thoracoscopic surgery (R-VATS) lobectomy for non-small cell lung cancer (NSCLC) over 36 months. Perioperative outcomes, tumor histology, numbers, locations, and status of all lymph nodes (LNs), and TNM (tumor, nodal, and metastasis) stage changes were analyzed. RESULTS: One hundred fifty-nine pts had mean tumor size 3.3±0.2 cm, most commonly being adenocarcinoma. Assessment of ≥3 N2 stations occurred in 156 (98.1%) pts, with 141 (88.7%) pts having >3 N2 stations reported. Mean total N1 + N2 stations assessed was 5.6±0.1 stations, including mean 4.1±0.1 N2 stations assessed. Mean N2 LNs reported was 7.2±0.3 LNs, and mean total N1 + N2 LNs reported was 13.4±0.4 LNs. There were 118 (74.2%) clinical stage-I pts versus 96 (60.4%) pathologic stage-I pts. Overall, 48 (30.2%) pts were upstaged, including 13 pts with cN0-pN1, 13 pts with cN0-pN2, 4 pts with cN1-pN2, and 18 pts with changes in T. CONCLUSIONS: R-VATS lobectomy is safe and results in perioperative outcomes comparable to those reported for conventional VATS. R-VATS MLND is effective at detecting occult metastatic disease during lobectomy for NSCLC.

Surgical outcomes associated with postoperative atrial fibrillation after robotic-assisted pulmonary lobectomy: retrospective review of 208 consecutive cases.

BACKGROUND: In this study, we sought to investigate the effect of post-operative atrial fibrillation (POAF) after robotic-assisted video-thoracoscopic pulmonary lobectomy on comorbid postoperative complications, chest tube duration, and hospital length of stay (LOS). METHODS: We retrospectively analyzed prospectively collected data from 208 consecutive patients who underwent robotic-assisted pulmonary lobectomy by one surgeon for known or suspected lung cancer. Postoperatively, 39 (18.8%) of these patients experienced POAF during their hospital stay. The occurrence of postoperative complications other than POAF, chest tube duration, and hospital LOS were analyzed in patients with POAF and without POAF. Statistical significance (P≤0.05) was determined by unpaired Student's t-test or by Chi-square test. RESULTS: Of patients with POAF, 46% also had other concurrent postoperative complications, while only 31% of patients without POAF experienced complications. The average number of postoperative complications experienced by patients with POAF was significantly higher than that experienced by those without POAF (0.9 vs. 0.4, P<0.05). Median chest tube duration in POAF patients (6 days) was significantly higher than in patients without POAF (4 days). A similar result was also seen with hospital LOS, with the median hospital LOS of 8 days in POAF patients being significantly longer than in those without POAF, whose median hospital LOS was 4 days. No other significant difference was detected between the two groups of patients. CONCLUSIONS: This study demonstrated the association between the incidence of POAF and a more complicated hospital course. Further studies are needed to determine whether confounders were involved in this association.

Domestic cats seropositive for Felis catus gammaherpesvirus 1 are often qPCR negative.

Felis catus gammaherpesvirus 1 (FcaGHV1) is a newly described virus that infects domestic cats. To identify FcaGHV1 antigens, we developed an immunofluorescent antibody assay by expressing FcaGHV1 open reading frames (ORFs) in feline cells and incubating fixed cells with sera from FcaGHV1-positive cats. Of the seven ORFs tested, ORF52 and ORF38 had the strongest, most consistent antibody responses. We used recombinant ORF52 and ORF38 proteins to develop two FcaGHV1 ELISAs. These assays were used to detect reactivity in cats previously tested by qPCR for FcaGHV1 in blood cell DNA. Results indicated 32%FcaGHV1seroprevalence, compared to 15%qPCR-evaluated prevalence (n=133);all but one qPCR positive animal was seropositive. ELISA results confirmed infection risk factors previously identified by qPCR: geographic location, male sex, and adult age. These data suggest that FcaGHV1is a common infection of domestic cats that has a seropositive but often qPCR negative state characteristic of herpesviral latency.

Effect of small body habitus on peri-operative outcomes after robotic-assisted pulmonary lobectomy: retrospective analysis of 208 consecutive cases.

BACKGROUND: Patients with smaller body surface area (BSA) have smaller pleural cavities, which limit visualization and instrument mobility during video-assisted thoracoscopic surgery (VATS). We investigated the effects of BSA on outcomes with robotic-assisted VATS lobectomy. METHODS: We analyzed 208 consecutive patients who underwent robotic-assisted lobectomy over 34 months. Patients were separated into group A (BSA ≤1.65 m(2)) and group B (BSA >1.65 m(2)). Operative times, estimated blood loss (EBL), conversions to thoracotomy, complications, hospital length of stay (LOS), and in-hospital mortality were compared. RESULTS: Group A had 40 patients (BSA 1.25-1.65 m(2)), and group B had 168 patients (BSA 1.66-2.86 m(2)). Median skin-to-skin operative times [± standard error of the mean (SEM)] were 169±16 min for group A and 176±6 min for group B (P=0.34). Group A had median EBL of 150±96 mL compared to 200±24 mL for group B (P=0.37). Overall conversion rate to thoracotomy was 8/40 (20.0%) in group A versus 12/168 (7.1%) in group B (P=0.03); while emergent conversion for bleeding was 2/40 (5.0%) in group A versus 5/168 (3.0%) in group B (P=0.62). Postoperative complications occurred in 12/40 (30.0%) in group A, compared to 66/168 (39.3%) in group B (P=0.28). Patients from both groups had median hospital LOS of 5 days (P=0.68) and had similar in-hospital mortality. CONCLUSIONS: Patients with BSA ≤1.65 m(2) have similar perioperative outcomes and complication risks as patients with larger BSA. Patients with BSA ≤1.65 m(2) have a higher overall conversion rate to thoracotomy, but similar conversion rate for bleeding as patients with larger BSA. Robotic-assisted pulmonary lobectomy is feasible and safe in patients with small body habitus.

Closing the gap on causal processes of infection risk from cross-sectional data: structural equation models to understand infection and co-infection.

BACKGROUND: Epidemiological studies of disease exposure risk are frequently based on observational, cross-sectional data, and use statistical approaches as crucial tools for formalising causal processes and making predictions of exposure risks. However, an acknowledged limitation of traditional models is that the inferred relationships are correlational, cannot easily distinguish direct from indirect determinants of disease risk, and are often considerable simplifications of complex interrelationships. This may be particularly important when attempting to infer causality in patterns of co-infection through pathogen-facilitation. METHODS: We describe analyses of cross-sectional data using structural equation models (SEMs), a contemporary advancement on traditional regression approaches, based on our study system of feline gammaherpesvirus (FcaGHV1) in domestic cats. RESULTS: SEMs strongly supported a latent (host phenotype) variable associated with FcaGHV1 exposure and co-infection risk, suggesting these individuals are simply more likely to become infected with multiple pathogens. However, indications of pathogen-covariance (potential facilitation) were also variably detected: potentially among FcaGHV1, Bartonella spp and Mycoplasma spp. CONCLUSIONS: Our models suggest multiple exposures are primarily driven by host phenotypic traits, such as aggressive male phenotypes, and secondarily by pathogen-pathogen interactions. The results of this study demonstrate the application of SEMs to understanding epidemiological processes using observational data, and could be used more widely as a complementary tool to understand complex cross-sectional information in a wide variety of disciplines.

Felis catus gammaherpesvirus 1; a widely endemic potential pathogen of domestic cats.

Felis catus gammaherpesvirus 1 (FcaGHV1), recently discovered in the USA, was detected in domestic cats in Australia (11.4%, 95% confidence interval 5.9-19.1, n=110) and Singapore (9.6%, 95% confidence interval 5.9-14.6, n=176) using qPCR. FcaGHV1 qPCR positive cats were 2.8 times more likely to be sick than healthy. Risk factors for FcaGHV1 detection included being male, increasing age and coinfection with pathogenic retroviruses, feline immunodeficiency virus (FIV) or feline leukaemia virus. FcaGHV1 DNA was detected in multiple tissues from infected cats with consistently high virus loads in the small intestine. FcaGHV1 viral load was significantly higher in FIV-infected cats compared with matched controls, mimicking increased Epstein-Barr virus loads in human immunodeficiency virus-infected humans. FcaGHV1 is endemic in distant geographic regions and is associated with being sick and with coinfections. Horizontal transmission of FcaGHV1 is supported, with biting being a plausible route. A pathogenic role for FcaGHV1 in domestic cats is supported.

Novel gammaherpesviruses in North American domestic cats, bobcats, and pumas: identification, prevalence, and risk factors.

UNLABELLED: Gammaherpesviruses (GHVs) are a diverse and rapidly expanding group of viruses associated with a variety of disease conditions in humans and animals. To identify felid GHVs, we screened domestic cat (Felis catus), bobcat (Lynx rufus), and puma (Puma concolor) blood cell DNA samples from California, Colorado, and Florida using a degenerate pan-GHV PCR. Additional pan-GHV and long-distance PCRs were used to sequence a contiguous 3.4-kb region of each putative virus species, including partial glycoprotein B and DNA polymerase genes. We identified three novel GHVs, each present predominantly in one felid species: Felis catus GHV 1 (FcaGHV1) in domestic cats, Lynx rufus GHV 1 (LruGHV1) in bobcats, and Puma concolor GHV 1 (PcoGHV1) in pumas. To estimate infection prevalence, we developed real-time quantitative PCR assays for each virus and screened additional DNA samples from all three species (n = 282). FcaGHV1 was detected in 16% of domestic cats across all study sites. LruGHV1 was detected in 47% of bobcats and 13% of pumas across all study sites, suggesting relatively common interspecific transmission. PcoGHV1 was detected in 6% of pumas, all from a specific region of Southern California. The risk of infection for each host varied with geographic location. Age was a positive risk factor for bobcat LruGHV1 infection, and age and being male were risk factors for domestic cat FcaGHV1 infection. Further characterization of these viruses may have significant health implications for domestic cats and may aid studies of free-ranging felid ecology. IMPORTANCE: Gammaherpesviruses (GHVs) establish lifelong infection in many animal species and can cause cancer and other diseases in humans and animals. In this study, we identified the DNA sequences of three GHVs present in the blood of domestic cats (Felis catus), bobcats (Lynx rufus), and pumas (Puma concolor; also known as mountain lions, cougars, and panthers). We found that these viruses were closely related to, but distinct from, other known GHVs of animals and represent the first GHVs identified to be native to these feline species. We developed techniques to rapidly and specifically detect the DNA of these viruses in feline blood and found that the domestic cat and bobcat viruses were widespread across the United States. In contrast, puma virus was found only in a specific region of Southern California. Surprisingly, the bobcat virus was also detected in some pumas, suggesting relatively common virus transmission between these species. Adult domestic cats and bobcats were at greater risk for infection than juveniles. Male domestic cats were at greater risk for infection than females. This study identifies three new viruses that are widespread in three feline species, indicates risk factors for infection that may relate to the route of infection, and demonstrates cross-species transmission between bobcats and pumas. These newly identified viruses may have important effects on feline health and ecology.

Stress and sociocultural factors related to health status among US-Mexico border farmworkers.

This study examines factors relating to farmworkers' health status from sociocultural factors, including stress embedded within their work and community contexts. A cross-sectional household survey of farmworkers (N = 299) included social-demographics, immigration status descriptors, and a social-ecologically grounded, community-responsive, stress assessment. Outcomes included three standard US national surveillance measures of poor mental, physical, and self-rated health (SRH). Logistic regression models showed that higher levels of stress were significantly associated (Ps < .001) with increased risk for poor mental health and poor physical health considering all variables. Stress was not associated with SRH. Regarding two of the three outcomes, mental health and physical health, stress added explanatory power as expected. For poor SRH, a known marker for mortality risk and quite high in the sample at 38%, only age was significantly associated. Clinical and systems-level health promotion strategies may be required to mitigate these stressors in border-residing farmworkers.

Minimally invasive mediastinal staging of non-small-cell lung cancer: emphasis on ultrasonography-guided fine-needle aspiration.

BACKGROUND: Mediastinal staging in patients with non-small-cell lung cancer (NSCLC) is crucial in dictating surgical vs nonsurgical treatment. Cervical mediastinoscopy is the "gold standard" in mediastinal staging but is invasive and limited in assessing the posterior subcarinal, lower mediastinal, and hilar lymph nodes. Less invasive approaches to NSCLC staging have become more widely available. METHODS: This article reviews several of these techniques, including noninvasive mediastinal staging of NSCLC, endobronchial ultrasound (EBUS) and fine-needle aspiration (FNA), endoscopic ultrasound (EUS) and FNA, and the combination of EBUS/EUS. RESULTS: Noninvasive mediastinal staging with computed tomography and positron-emission tomography scans has significant false-negative and false-positive rates and requires lymph node tissue confirmation. FNA techniques, with guidance by EBUS and EUS, have become more widely available. The combination of EBUS-FNA and EUS-FNA of mediastinal lymph nodes can be a viable alternative to surgical mediastinal staging. Current barriers to the dissemination of these techniques include initial cost of equipment, lack of access to rapid on-site cytology, and the time required to obtain sufficient skills to duplicate published results. CONCLUSIONS: Within the last decade, these approaches to NSCLC staging have become more widely available. Continued study into these noninvasive techniques is warranted.

Neuronal nitric oxide synthase is phosphorylated in response to insulin stimulation in skeletal muscle.

Type 2 Diabetes (T2DM) is the seventh leading cause of death in the United States, and is quickly becoming a global pandemic. T2DM results from reduced insulin sensitivity coupled with a relative failure of insulin secretion. Reduced insulin sensitivity has been associated with reduced nitric oxide synthase (NOS) activity and impaired glucose uptake in T2DM skeletal muscle. Upon insulin stimulation, NO synthesis increases in normal adult skeletal muscle, whereas no such increase is observed in T2DM adults. Endothelial NOS is activated by phosphorylation in the C-terminal tail in response to insulin. Neuronal NOS (nNOS), the primary NOS isoform in skeletal muscle, contains a homologous phosphorylation site, raising the possibility that nNOS, too, may undergo an activating phosphorylation event upon insulin treatment. Yet it remains unknown if or how nNOS is regulated by insulin in skeletal muscle. Data shown herein indicate that nNOS is phosphorylated in response to insulin in skeletal muscle and that this phosphorylation event occurs rapidly in C2C12 myotubes, resulting in increased NO production. In vivo phosphorylation of nNOS was also observed in response to insulin in mouse skeletal muscle. These results indicate, for the first time, that nNOS is phosphorylated in skeletal muscle in response to insulin and in association with increased NO production.

Persons with Parkinson's disease exhibit decreased neuromuscular complexity during gait.

OBJECTIVE: Individual muscle activation patterns may be controlled by motor modules constructed by the central nervous system to simplify motor control. This study compared modular control of gait between persons with Parkinson's disease (PD) and neurologically-healthy older adults (HOA) and investigated relationships between modular organization and gait parameters in persons with PD. METHODS: Fifteen persons with idiopathic PD and fourteen HOA participated. Electromyographic recordings were made from eight leg muscles bilaterally while participants walked at their preferred walking speed for 10 min on an instrumented treadmill. Non-negative matrix factorization techniques decomposed the electromyographic signals, identifying the number and nature of modules accounting for 95% of variability in muscle activations during treadmill walking. RESULTS: Generally, fewer modules were required to reconstruct muscle activation patterns during treadmill walking in PD compared to HOA (p < .05). Control of knee flexor and ankle plantar flexor musculature was simplified in PD. Activation timing was altered in PD while muscle weightings were unaffected. Simplified neuromuscular control was related to decreased walking speed in PD. CONCLUSION: Neuromuscular control of gait is simplified in PD and may contribute to gait deficits in this population. SIGNIFICANCE: Future studies of locomotor rehabilitation in PD should consider neuromuscular complexity to maximize intervention effectiveness.