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This study aimed to evaluate the frequency of sleep and eating problems in children with autism spectrum disorder (ASD), the factors associated with these problems, and the relationship with the risk of developing depression and anxiety in their parents. In the study, 156 children with ASD and their parents were included. The Brief Autism Mealtime Behavior Inventory (BAMBI), the Children's Sleep Habits Questionnaire (CSHQ), the Hospital Anxiety Depression Scale (HADS) were completed by the parents. The Childhood Autism Rating Scale (CARS) was administered by the researchers to evaluate the severity of ASD in children. The mean age of children was 8.08 ± 2.84, and 87.2% of the children were male. The mean score of CARS was 41.25 ± 6.16, the mean score of BAMBI was 45.67 ± 11.87, and the mean score of CSHQ was 51.67 ± 10.17. According to the CSHQ, 83.3% of the children had sleep problems. The response rate above the cut-off point in the HAD-A subscale was 59.6%, whereas it was 67.3% in the HAD-D subscale. There was a positive correlation between CARS and BAMBI, CSHQ, HAD-A, and HAD-D subscales. There was a positive correlation between BAMBI and CHSQ, HAD-A, and HAD-D subscales. There was a positive correlation between the CSHQ, HAD-A, and HAD-D subscales. There was a negative correlation between the age of the child with ASD, food refusal, and resistance to bedtime. As sleeping and eating problems affect a large portion of children diagnosed with ASD and their caregivers, large population-based studies evaluating both these problems must be designed and carried out to understand factors affecting the prevalence, development, and persistence of sleeping and eating problems and to determine interventions to reduce these issues.

Current evidence shows that early interventions in children with autism spectrum disorder (ASD) are more efficient and associated with better long-term outcomes. This also highlights the need for early recognition of children with ASD symptoms by family physicians and referral to child psychiatrists. This study aimed to evaluate how family physicians’ knowledge about ASD is reflected in clinical practice by making a diagnostic evaluation of the cases referred with suspected ASD. The study included 149 cases that family physicians referred to child psychiatrists regarding the assessment for ASD. Diagnostic evaluation of the subjects was carried out using the Diagnostic and Statistical Manual of Mental Disorders, fifth edition (DSM-5) criteria. Childhood Autism Rating Scale (CARS) and Autism Behavior Checklist (ABC) were used in the evaluation process. Denver II developmental screening inventory was used to evaluate all subjects under six. The mean age of all participants was 39.1 (±13.2) months, and in ASD cases, it was 34.6 (±7.8) months. Of all referred cases, 72,5% were diagnosed according to the DSM-5 criteria. The most common diagnoses were global developmental delay (28.9%), language disorder (15.4%), ASD (15.4%), and attention deficit hyperactivity disorder (8.7%), respectively. The most common reason for referral specified on the consultation note was delayed speech, with a rate of 87.2%. In addition, 148 (99.3%) of the symptoms noted on the referral note were clustered within the DSM-5 ASD A criteria and 19 (12.8%) within the ASD B criteria. Developmental delay was more prominent in cases with ASD when compared to those with global developmental delay and language disorder. The most common diagnosis was a global developmental delay in referred cases, and only some of the symptoms seen in ASD from the referral notes of family physicians stood out as the reason for referral. These results underline the necessity of continuing ASD training for family physicians.

Background: Atypical hemolytic syndrome (aHUS) and C3 glomerulopathy (C3G) are complement-mediated rare diseases with excessive activation of the alternative pathway. Data to guide the evaluation of living-donor candidates for aHUS and C3G are very limited. The outcomes of living donors to recipients with aHUS and C3G (Complement disease-living donor group) were compared with a control group to improve our understanding of the clinical course and outcomes of living donation in this context. Methods: Complement disease-living donor group [n= 28; aHUS(53.6%), C3G(46.4%)] and propensity score-matched control-living donor group (n= 28) were retrospectively identified from 4 centers (2003–2021) and followed for major cardiac events (MACE), de novo hypertension, thrombotic microangiopathy (TMA), cancer, death, estimated glomerular filtration rate (eGFR) and proteinuria after donation. Results: None of the donors for recipients with complement-related kidney diseases experienced MACE or TMA whereas two donors in the control group developed MACE (7.1%) after 8 (IQR, 2.6–12.8) years (p= 0.15). New-onset hypertension was similar between complement disease and control donor groups (21.4% vs 25%, respectively, p= 0.75). There were no differences between study groups regarding last eGFR and proteinuria levels (p= 0.11 and p= 0.70, respectively). One related donor for a recipient with complement-related kidney disease developed gastric cancer and another related donor developed a brain tumor and died in the 4th year after donation (2, 7.1% vs none, p= 0.15). No recipient had donor-specific human leukocyte antigen antibodies at the time of transplantation. Median follow-up period of transplant recipients was 5 years (IQR, 3–7). Eleven (39.3%) recipients [aHUS (n = 3) and C3G (n = 8)] lost their allografts during the follow-up period. Causes of allograft loss were chronic antibody-mediated rejection in 6 recipients and recurrence of C3G in 5. Last serum creatinine and last eGFR of the remaining patients on follow up were 1.03 ± 038 mg/dL and 73.2 ± 19.9 m/min/1.73 m2 for aHUS patients and 1.30 ± 0.23 mg/dL and 56.4 ± 5.5 m/min/1.73 m2 for C3G patients. Conclusion: The present study highlights the importance and complexity of living related-donor kidney transplant for patients with complement-related kidney disorders and motivates the need for further research to determine the optimal risk-assessment for living donor candidates to recipients with aHUS and C3G. Graphical abstract:

Insulin is one of the major hormones involved in the control of energy expenditure and carbohydrate, lipid, and protein metabolism. It also regulates a variety of biological processes such as [...], OBJECTIVE--Factors governing microRNA expressions in response to changes of cellular environment are still largely unknown. Our aim was to determine whether insulin, the major hormone controlling whole-body energy homeostasis, is involved in the regulation of microRNA expressions in human skeletal muscle. RESEARCH DESIGN AND METHODS--We carried out comparative microRNA (miRNA) expression profiles in human skeletal muscle biopsies before and after a 3-h euglycemichyperinsulinemic clamp, with TaqMan low-density arrays. Then, using DNA microarrays, we determined the response to insulin of the miRNA putative target genes in order to determine their role in the transcriptional action of insulin. We further characterized the mechanism of action of insulin on two representative miRNAs, miR-1 and miR-133a, in human muscle cells. RESULTS--Insulin downregulated the expressions of 39 distinct miRNAs in human skeletal muscle. Their potential target mRNAs coded for proteins that were mainly involved in insulin signaling and ubiquitination-mediated proteolysis. Bioinformatic analysis suggested that combinations of different downregulated miRNAs worked in concert to regulate gene expressions in response to insulin. We further demonstrated that sterol regulatory element-binding protein (SREBP)-1c and myocyte enhancer factor 2C were involved in the effect of insulin on miR-1 and miR-133a expression. Interestingly, we found an impaired regulation of miRNAs by insulin in the skeletal muscle of type 2 diabetic patients, likely as consequences of altered SREBP-1c activation. CONCLUSIONS--This work demonstrates a new role of insulin in the regulation of miRNAs in human skeletal muscle and suggests a possible implication of these new modulators in insulin resistance.

This study aimed to investigate the emotional and behavioral responses of children with neurodevelopmental disorders and their parents during the recent novel coronavirus disease (COVID-19) pandemic and the associated factors. Our study included 299 children and adolescents with neurodevelopmental disorders and 299 mothers or fathers. Participant groups were as follows: autism spectrum disorder (n = 131, 43.8%); intellectual disability (n = 103, 34.4%); specific learning disorder (n = 46, 15.4%); and communication disorder (n = 19, 6.4%). A data form was created to investigate the emotional, behavioral, sleep problems, and appetite changes of the cases, and it was adapted for online application and conveyed to the parents in the form of emails. Our study indicated that children and adolescents’ emotional problems were 44.5%, behavioral problems 33.4%, sleep problems 65.2%, and appetite problems 32.4%. Irritability (35.5%), hyperactivity (37.8%), stereotyped behavior (29.8%), delayed sleep phase (51.5%) were the most common problems reported by the parents. The frequencies of emotional and behavioral problems in the ASD group were higher than in the other groups (p < .05). In addition, the Hospital Anxiety Depression Scale (HADS) was used to determine the parents’ anxiety and depression symptoms. Mean HADS anxiety and depression scores of parents were 8.5 ± 4.3 and 7.8 ± 4.2, respectively. Our study demonstrated that the COVID-19 pandemic and lockdown negatively affected children with neurodevelopmental disorders and their parents in many areas. It is necessary that these children and their families are evaluated and supported by local professionals, as the ongoing pandemic can exacerbate the current challenges. The findings of our study reflect the early period of the pandemic. For this reason, longitudinal studies about the long-term consequences of the pandemic are needed.

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.ajog.2006.01.067 Byline: Nikki Koklanaris (a), Jerome C. Nwachukwu (b), S. Joseph Huang (c), Seth Guller (c), Ksenia Karpisheva (b), Michael Garabedian (b), Men-Jean Lee (c) Abstract: Trophoblast invasion, which sets the stage for placentation and pregnancy outcome, likely occurs in a hypoxic environment. We used microarray technology in a trophoblast cell line to identify hypoxia-responsive genes that may impact placentation. Author Affiliation: (a) Department of Obstetrics and Gynecology, Division of Maternal-Fetal Medicine (b) Department of Microbiology, New York University School of Medicine, New York, NY (c) Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, New Haven, CT Article History: Received 8 December 2005; Revised 9 January 2006; Accepted 17 January 2006 Article Note: (footnote) Presented at the 26th Annual Meeting of the Society for Maternal Fetal Medicine, January 30-February 4, 2006, Miami, FL.

The clustering of hypertensive disorders due to the alterations in urinary levels of vascular endothelial growth factor (VEGF), tyrosine kinase 1 (sFlt-1) and placental growth factor (PIGF) is examined and women with severe preeclampsia are identified. At the time of clinical manifestation, it is observed that severe preeclampsia (sPE) is associated with increased urinary output of the antiangiogenic factor sFlt-1 and a decreased output of PIGF.

IMPORTANCE: Tumor histological factors that predict immunotherapy response in patients with recurrent or metastatic head and neck squamous cell carcinoma (HNSCC) are not well defined. OBJECTIVE: To investigate the association between tumor grade and immunotherapy response in patients with recurrent or metastatic mucosal HNSCC. DESIGN, SETTING, AND PARTICIPANTS: In this retrospective cohort study, the medical records of 60 patients with recurrent or metastatic mucosal HNSCC treated with immune checkpoint inhibitors at Johns Hopkins Hospital between July 1, 2015, and January 22, 2020, were reviewed. EXPOSURES: High-grade tumors (HGTs) vs low-grade tumors (LGTs) in recurrent or metastatic HNSCC. MAIN OUTCOMES AND MEASURES: Patients were divided into 2 groups: those with LGTs (well differentiated and moderately differentiated) and those with HGTs (poorly differentiated). The main outcome was a clinically beneficial immunotherapy response, defined as complete response or partial response. Univariable and multivariable logistic regressions were conducted to calculate odds ratios for each variable’s association with immunotherapy response. Survival differences were evaluated using Kaplan-Meier survival curves with multivariable Cox proportional hazards regression models. RESULTS: The 60 patients (35 with HGTs and 25 with LGTs) had a mean (SD) age of 64.6 (8.88) years; 51 were male (85%); and 38 were current or former smokers (63%). The oropharynx was the most common primary tumor site both in patients with HGTs (22 of 35; 63%) and those with LGTs (12 of 25; 48%). Bivariate analysis showed the proportion of patients having a beneficial response to immunotherapy was greater for patients with HGTs (12 of 35; 34.3%) than those with LGTs (2 of 25, 8.0%) (difference, 26.3%; 95% CI, 7.3%-45.3%). Upon multivariable analysis, patients with HGTs had 5.35-fold increased odds (95% CI, 1.04-27.37) of having a clinically beneficial response to immunotherapy. Among patients with available tumor genomic profiling data, the mean tumor mutational burden was greater for patients with HGTs (mean [SD], 8.6 [5.4] mut/Mb; n = 8) than patients with LGTs (mean [SD], 3.6 [1.1] mut/Mb; n = 4) (difference = 5.0 mut/Mb; 95% CI −1.4 to 11.4 mut/Mb; Cohen d = 1.2). CONCLUSIONS AND RELEVANCE: In this cohort study, tumor grade was independently associated with immunotherapy response in patients with recurrent or metastatic mucosal HNSCC. These findings highlight the potential role of tumor grade in predicting immunotherapy response in mucosal HNSCC.

Immunoglobulin G (IgG) in patients with the antiphospholipid syndrome appears to inhibit the activity of a powerful anti-blood-clotting protein called annexin V. This could account for the increased incidence of miscarriage among pregnant women with this syndrome. Annexin V levels were lower in placental tissue from a patient with the antiphospholipid syndrome than in a healthy mother. Annexin V production decreased in normal placental tissue treated with IgG from patients with the antiphospholipid syndrome.

IMPORTANCE: Human papillomavirus (HPV)–positive status in patients with oropharyngeal squamous cell carcinoma (OPSCC) is associated with improved survival compared with HPV-negative status. However, it remains controversial whether HPV is associated with improved survival among patients with nonoropharyngeal and cervical squamous cell tumors. OBJECTIVE: To investigate differences in the immunogenomic landscapes of HPV-associated tumors across anatomical sites (the head and neck and the cervix) and their association with survival. DESIGN, SETTING, AND PARTICIPANTS: This cohort study used genomic and transcriptomic data from the Cancer Genome Atlas (TCGA) for 79 patients with OPSCC, 435 with nonoropharyngeal head and neck squamous cell carcinoma (non-OP HNSCC), and 254 with cervical squamous cell carcinoma and/or endocervical adenocarcinoma (CESC) along with matched clinical data from TCGA. The data were analyzed from November 2020 to March 2021. MAIN OUTCOMES AND MEASURES: Positivity for HPV was classified by RNA-sequencing reads aligned with the HPV reference genome. Gene expression profiles, immune cell phenotypes, cytolytic activity scores, and overall survival were compared by HPV tumor status across multiple anatomical sites. RESULTS: The study comprised 768 patients, including 514 (66.9%) with HNSCC (380 male [73.9%]; mean [SD] age, 59.5 [10.8] years) and 254 (33.1%) with CESC (mean [SD] age, 48.7 [14.1] years). Human papillomavirus positivity was associated with a statistically significant improvement in overall survival for patients with OPSCC (adjusted hazard ratio [aHR], 0.06; 95% CI, 0.02-0.17; P &lt; .001) but not for those with non-OP HNSCC (aHR, 0.64; 95% CI, 0.31-1.27; P = .20) or CESC (aHR, 0.50; 95% CI, 0.15-1.67; P = .30). The HPV-positive OPSCCs had increased tumor immune infiltration and immunomodulatory receptor expression compared with HPV-negative OPSCCs. Compared with HPV-positive non-OP HNSCCs, HPV-positive OPSCCs showed greater expression of immune-related metrics including B cells, T cells, CD8+ T cells, T-cell receptor diversity, B-cell receptor diversity, and cytolytic activity scores, independent of tumor variant burden. The immune-related metrics were similar when comparing HPV-positive non-OP HNSCCs and HPV-positive CESCs with their HPV-negative counterparts. The 2-year overall survival rate was significantly higher for patients with HPV-positive OPSCC compared with patients with HPV-negative OPSCC (92.0% [95% CI, 84.8%-99.9%] vs 45.8% [95% CI, 28.3%-74.1%]; HR, 0.10 [95% CI, 0.03-0.30]; P = .009). CONCLUSIONS AND RELEVANCE: In this cohort study, tumor site was associated with the immune landscape and survival among patients with HPV-related tumors despite presumed similar biologic characteristics. These tumor site–related findings provide insight on possible outcomes of HPV positivity for tumors in oropharyngeal and nonoropharyngeal sites and a rationale for the stratification of HPV-associated tumors by site and the subsequent development of strategies targeting immune exclusion in HPV-positive nonoropharyngeal cancer.

Reduction of annexin-V in the placental villi appears a likely mechanism for recurrent miscarriage caused by the formation of placental clots among women with antiphospholipid antibodies (APA). Annexin-V is an anticoagulant produced in the placental villi that bonds to phospholipid proteins in the tips of the placental villi. It is believed to keep the blood in the maternal capillary sinuses fluid. The amount and location of annexin-V were evaluated in placental samples taken from eight placentas each from the following groups: healthy term births, women with APA and recurrent miscarriage, women with miscarriage from other causes, and women undergoing elective abortion. Normal placental villi were also cultured with APA. Placentas from women with APA had much less annexin-V in the villi tips compared with the other groups. It remained present deeper in the villous cells. The same results were seen in normal villi cultured in the presence of APA.

This case report describes a unique transmission of Vibrio vulnificus infection. A 38-year-old woman with recurrent cellulitis and chronic ulcer on her leg developed necrotizing cellulitis and sepsis caused by [...]

A hereditary cancer syndrome is a genetic predisposition to cancer caused by a germline mutation in cancer-related genes. Identifying the disease-causing variant is important for both the patient and relatives at risk in cancer families because this could be a guide in treatment and secondary cancer prevention. In this study, hereditary cancer panel harboring cancer-related genes was performed on MiSeq Illumina NGS system from peripheral blood samples. Sequencing files were fed into a cloud-based data analysis pipeline. Reportable variants were classified according to the American College of Medical Genetics and Genomics guidelines. Three hundred five individuals were included in the study. Different pathogenic/likely pathogenic variants were detected in 75 individuals. The majority of these variants were in the MUTYH, BRCA2, and CHEK2genes. Nine novel pathogenic/likely pathogenic variants were identified in BRCA1, BRCA2, GALNT12, ATM, MLH1, MSH2, APC, and KITgenes. We obtained interesting and novel variants which could be related to hereditary cancer, and this study confirmed that NGS is an indispensable method for the risk assessment in cancer families.

Decidual macrophages are in close contact with trophoblast cells during placenta development, and an appropriate crosstalk between these cellular compartments is crucial for the establishment and maintenance of a healthy pregnancy. During different phases of gestation, macrophages undergo dynamic changes to adjust to the different stages of fetal development. Trophoblast‐secreted factors are considered the main modulators responsible for macrophage differentiation and function. However, the phenotype of these macrophages induced by trophoblast‐secreted factors and the factors responsible for their polarization has not been elucidated. In this study, we characterized the phenotype and function of human trophoblast‐induced macrophages. Using in vitro models, we found that human trophoblast‐educated macrophages were CD14+CD206+CD86−and presented an unusual transcriptional profile in response to TLR4/LPS activation characterized by the expression of type I IFN‐β expression. IFN‐β further enhances the constitutive production of soluble programmed cell death ligand 1 (PD‐L1) from trophoblast cells. PD‐1 blockage inhibited trophoblast‐induced macrophage differentiation. Soluble PD‐L1 (sPD‐L1) was detected in the blood of pregnant women and increased throughout the gestation. Collectively, our data suggest the existence of a regulatory circuit at the maternal fetal interface wherein IFN‐β promotes sPD‐L1 expression/secretion by trophoblast cells, which can then initiate a PD‐L1/PD‐1‐mediated macrophage polarization toward an M2 phenotype, consequently decreasing inflammation. Macrophages then maintain the expression of sPD‐L1 by the trophoblasts through IFN‐β production induced through TLR4 ligation. Trophoblast‐secreted factors resulted in macrophages differentiation, leading to unique phenotypes and differences in Toll Like receptor responses to bacterial products.

In this work, we brought together two existing clinical techniques used in cancer treatment—X-ray radiation and photodynamic therapy (PDT), whose combination termed X-PDT uniquely allows PDT to be therapeutically effective in deep tissue. To this end, we developed mitochondrially targeted biodegradable polymer poly(lactic-co-glycolic acid) nanocarriers incorporating a photosensitizer verteporfin, ultrasmall (2–5 nm) gold nanoparticles as radiation enhancers, and triphenylphosphonium acting as the mitochondrial targeting moiety. The average size of the nanocarriers was about 160 nm. Upon X-ray radiation our nanocarriers generated cytotoxic amounts of singlet oxygen within the mitochondria, triggering the loss of membrane potential and mitochondria-related apoptosis of cancer cells. Our X-PDT strategy effectively controlled tumor growth with only a fraction of radiotherapy dose (4 Gy) and improved the survival rate of a mouse model bearing colorectal cancer cells. In vivodata indicate that our X-PDT treatment is cytoreductive, antiproliferative, and profibrotic. The nanocarriers induce radiosensitization effectively, which makes it possible to amplify the effects of radiation. A radiation dose of 4 Gy combined with our nanocarriers allows equivalent control of tumor growth as 12 Gy of radiation, but with greatly reduced radiation side effects (significant weight loss and resultant death).

Introduction: People experiencing unsheltered homelessness (PEUH) have higher disease burden yet limited access to healthcare. COVID-19 introduced even greater risk for PEUH aged 65+ years with an underlying chronic health condition and were temporarily housed in hotels/motels for Project RoomKey (PRK). This study aimed to characterize a PRK cohort who received primary care from a street medicine program.Methods: This observational case series study included a sample of 35 PRK participants receiving primary care from a street medicine team at a single site from July to September 2020. We used the HOUSED BEDS assessment tool for taking history on PEUH.Results: Participants were 63% male, 40% Hispanic/Latino/a, 40% white, 94% English-speaking, and 73% had chronic health conditions. Assessment revealed: average Homelessness (H) of 4 years; 76% had no prior social service Outreach (O); average Utilization (U) was 4 emergency department visits in prior 6-months; 68% received Salary (S) from government income; Food access or Eat (E) was commonly purchased (29%) or donated (26%); clean water to Drink (D) for 59% of participants; 86% had access to a Bathroom (B); Encampment (E) was varied and 38% reported safety concerns; Daily routine (D) showed 76% could access a telephone, 32% received social support from family; 79% reported past or current Substance use (S). No participants contracted COVID-19 during study period.Conclusions: This study describes health and demographic characteristics of PRK participants in Southern California. Findings inform policies to continue PRK that includes onsite healthcare such as via street medicine.

Background: While Thioredoxin Reductase (TrxR) plays an important role in regulation of the intracellular redox balance and various signalling pathways, Glutathione S-Transferase (GSTs) enzymes belong to the detoxification family that catalyse the conjugation of glutathione with various endogenous and xenobiotic electrophiles. Since TrxR and GSTs are overexpressed in many cancer cells, they have been identified as potential targets to develop chemotherapeutic strategies. Method: The mitochondrial TrxR (TrxR2) enzyme and the cytosolic GST enzyme was purified from rat liver via affinity chromatography. After the purification, the in vitro inhibition effects of some anticancer drugs (cisplatin, calcium folinate, carboplatin, epirubicin hydrochloride, doxorubicin hydrochloride, paclitaxel, etoposide, fluorouracil, and methotrexate) were investigated on both enzymes. Since only methotrexate inhibits both enzymes among all the anticancer drugs, a molecular docking study was performed to determine the binding site and the binding affinity of methotrexate to the enzymes. Results: Firstly, TrxR2 and GST were found to have a specific activity of 0.436, 1765 EU/mg proteins with a yield of 39.20%, 31.28% and 207.6, 3516.6 of purification fold, respectively. While TrxR2 was strongly inhibited by all of the anticancer drugs, GST was not inhibited by any of the anticancer drugs except methotrexate. Conclusion: Both enzymes were inhibited by only methotrexate in rat liver, and methotrexate was well placed in the active sites of both proteins. Therefore, it may be argued that methotrexate may be a more effective anticancer drug than all other drugs used in this study against the multi drug resistance that will occur during chemotherapy.

Upconversion nanoparticles (UCNPs) coated with polyethylenimine (PEI) are popular background-free optical contrast probes and efficient drug and gene delivery agents attracting attention in science, industry, and medicine. Their unique optical properties are especially useful for subsurface nanotheranostics applications, in particular, in skin. However, high cytotoxicity of PEI limits safe use of UCNP@PEI, and this represents a major barrier for clinical translation of UCNP@PEI-based technologies. Our study aims to address this problem by exploring additional surface modifications to UCNP@PEI to create less toxic and functional nanotheranostic materials. We designed and synthesized six types of layered polymer coatings that envelop the original UCNP@PEI surface, five of which reduced the cytotoxicity to human skin keratinocytes under acute (24 h) and subacute (120 h) exposure. In parallel, we examined the photoluminescence spectra and lifetime of the surface-modified UCNP@PEI. To quantify their brightness, we developed original methodology to precisely measure the colloidal concentration to normalize the photoluminescence signal using a nondigesting mass spectrometry protocol. Our results, specified for the individual coatings, show that, despite decreasing the cytotoxicity, the external polymer coatings of UCNP@PEI quench the upconversion photoluminescence in biologically relevant aqueous environments. This trade-off between cytotoxicity and brightness for surface-coated UCNPs emphasizes the need for the combined assessment of the viability of normal cells exposed to the nanoparticles and the photophysical properties of postmodification UCNPs. We present an optimized methodology for rational surface design of UCNP@PEI in biologically relevant conditions, which is essential to facilitate the translation of such nanoparticles to the clinical applications.

IMPORTANCE: Small nodal tumor infiltrates (SNTI; isolated tumor cells and micrometastases) in sentinel lymph nodes and bone marrow micrometastases (BMM) were independently described as prognostic factors in patients with colon cancer. OBJECTIVE: To examine the association between the occurrence of SNTI and BMM as well as their prognostic relevance. DESIGN, SETTING, AND PARTICIPANTS: This prospective study was conducted at 3 university-affiliated institutions in Switzerland between May 2000 and December 2006. Statistical analyses were performed in October 2016. A total of 122 patients with stage I to III colon cancer were included. Follow-up time exceeded 6 years, with no patients lost to follow-up. INTERVENTIONS: Bone marrow aspiration from the iliac crests and in vivo sentinel lymph node mapping were performed during open standard oncological resection. Bone marrow aspirates were stained with the pancytokeratin marker A45-B/B3. All sentinel lymph nodes underwent multilevel sectioning and were stained with hematoxylin-eosin and the pancytokeratin marker AE1/AE3. MAIN OUTCOMES AND MEASURES: Association of SNTI in sentinel lymph nodes and BMM in patients with stage I to III colon cancer and the prognostic effect on disease-free survival (DFS) and overall survival (OS). RESULTS: Of the 122 patients, 63 (51.6%) were female, with a mean (SD) age of 71.2 (11.7) years. Small nodal tumor infiltrates and BMM were found in a total of 21 patients (17.2%) and 46 patients (37.7%), respectively. The occurrence of BMM was not associated with the presence of SNTI by standard correlation (κ, −0.07; 95% CI, −0.29 to 0.14; P = .49) nor by univariate logistic regression analysis (odds ratio, 0.64; 95% CI, 0.22-1.67; P = .37) or multivariate logistic regression analysis (odds ratio, 1.09; 95% CI, 0.34-3.28; P = .88). The presence of SNTI was an independent negative prognostic factor for DFS (hazard ratio [HR], 2.93; 95% CI, 1.24-6.93; P = .02) and OS (HR, 4.04; 95% CI, 1.56-10.45; P = .005), as was BMM (HR, 2.07; 95% CI, 1.06-4.06; P = .04; and HR, 2.68; 95% CI, 1.26-5.70; P = .01; respectively). The combined detection of BMM and SNTI demonstrated the poorest DFS (HR, 6.73; 95% CI, 2.29-19.76; P = .006) and OS (HR, 5.96; 95% CI, 1.66-21.49; P = .03). CONCLUSIONS AND RELEVANCE: This study demonstrates no association between the occurrence of SNTI and BMM in patients with stage I to III colon cancer. However, both SNTI and BMM are independent negative prognostic factors regarding DFS and OS, and the occurrence of both is associated with significantly worse prognosis compared with either one of them. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00826579

The model established at Orillia Soldiers Memorial Hospital involves family physicians as the most responsible physician. They act as “admission gatekeeper” for all unattached patients who are admitted to the psychiatry in-patient unit. A PubMed, EBSCO, OVID Medline, Embase, CINAHL, and Web of Science database review of the last 10 years (2006-2016) was undertaken. A satisfaction survey was undertaken. An intensive literature review found this model to be unique. The model has proved to be extremely efficient and cost-effective.

Discusses the use of various screening methods by correctional agencies to help assess new candidates for correctional positions. History of psychological screening for law enforcement personnel; Effectiveness of specialized psychological screening for new recruits at several prison systems in Australia; Overview of attrition rates and use of sick leave at New South Wales Department of Corrective Services.

The increasing range of surgery in elderly patients reflects the changing demography where in the next 10 years one quarter of the population will be 65 years of age or older. There is presently no consensus concerning the optimal predictive markers for postoperative morbidity and mortality after surgery in older patients with an appreciation that physical frailty is more important than chronological age. In this retrospective analysis, we have compared the impact of age and the calculated preoperative Charlson Comorbidity Index (CCI) on early (30-day) and late (one-year) mortality in a group of patients >75 years of age dividing them into an “older old” cohort (75–84 years of age, Group A) and an “oldest old” group (=85 years of age, Group B). Increased age was associated with a higher death rate after emergency surgery, with late deaths after elective surgery exceeding those after emergency operations. A higher mean CCI was noted in both age groups in early nonsurvivors after both elective and emergency surgery with a more significant effect of the preoperative CCI than chronological age for the prediction of late postoperative death for both groups after elective and emergency operations. Although the CCI was not designed to predict perioperative mortality in surgical cohorts, it correlates with a greater risk than age for perioperative death in the elderly.

Sustained plasma progesterone (P4) levels suggest initiation of human term labor by functional P4 withdrawal, reflecting reduced progesterone receptor (PR) and/or glucocorticoid receptor (GR) expression or activity. The steroid-induced immunophilin cochaperone FKBP51 inhibits PR- and GR-mediated transcription, suggesting a labor-initiating role. Gestational age–matched decidual sections were immunostained for FKBP51 and decidual cell (DC) and interstitial trophoblast (IT) markers, vimentin and cytokeratin, respectively. Term DC cultures were incubated with vehicle (control), estradiol (E2) with or without medroxyprogesterone acetate, dexamethasone (Dex), or Organon 2058. FKBP51 histologic scoring was significantly higher in DC nuclei during labor versus prelabor decidua, whereas FKBP51 immunostaining was undetected in interstitial trophoblasts (P < 0.05). In term DC cultures, E2 + medroxyprogesterone acetate or E2+ Dex enhanced FKBP51 expression (P < 0.01), whereas E2+ Organon 2058 inhibited PR expression (P < 0.05), and E2+ Dex inhibited GR expression (P < 0.05). Unlike term DCs, FKBP51 was undetected in control or Dex-treated cultured third-trimester trophoblasts. Electrophoretic mobility shift assays revealed that FKPB51 overexpression or silencing in cultured DCs altered PR-DNA binding. Increased FKBP51 levels in term DCs during labor complement our prior in situobservations of significantly lower PR in labor versus prelabor DCs. In a milieu of sustained plasma P4 levels, these reciprocal changes will amplify functional P4 withdrawal in DCs via FKBP51-mediated PR resistance coupled with declining PR levels, whereas the lack of FKBP51 expression in interstitial trophoblasts suggests unopposed constitutive GR action.

An 87-year-old man presented with a 3-day history of progressive, diffuse abdominal pain. He denied nausea, vomiting, fever or chills. He had no history of rectal bleeding, diarrhea or constipation, [...]

The Internet is an excellent avenue for businesses to market their products and services. Designing the proper Web site is important step to getting the messages out and making it easy for users to explore the options provided.

Patients with pancreatic adenocarcinoma often present with distant metastatic disease. We aimed to assess whether improvements in survival of clinical trials translated to a population-based level.

The aim of this study was to test two competing theories concerning the trait of urgency: (a) Urgency reflects the tendency to act rashly/impulsively when emotional, or (b) urgency reflects a general reflexive responsivity to emotions that can lead either to rash action or to ill-advised inaction and thus to either impulsive behavior or depression. Following prior findings that fifth-grade urgency predicted sixth-grade impulsive behavior, we tested whether urgency also predicted sixth-grade depression (n= 1,906). After controlling for sex, early pubertal onset, fifth-grade depression, fifth-grade engagement in addictive behaviors, negative affect, positive affect, and other impulsivity-related traits, fifth-grade urgency level did predict higher levels of depression at the end of sixth grade. This finding is consistent with the view that urgency can lead either to rash action or to ill-advised inaction. Urgency may be of transdiagnostic importance, contributing to both internalizing and externalizing dysfunction.

AbstractSchizophrenia is a severe mental illness with neurobiological bases that remain elusive. One hypothesis emphasizes disordered thalamic function. We previously used concurrent single pulse transcranial magnetic stimulation (spTMS) and functional magnetic resonance imaging (fMRI) to show that individuals with schizophrenia have a decreased spTMS-evoked response in the thalamus, and decreased effective connectivity between thalamus and insula and thalamus and superior frontal gyrus. To better understand the factors that may accompany or account for these findings, we investigated, in the same participants, resting state functional connectivity, white matter structural connectivity, and grey matter integrity. Patients with schizophrenia did not differ from healthy control subjects in resting state functional- or white matter structural connectivity, although they did show decreased measures of grey matter integrity in the insula. However, in this region, the spTMS-evoked response did not differ between groups. In a region of the thalamus that also had grey matter intensity abnormalities, although not at a level that survived correction for multiple comparisons, the spTMS-evoked response in patients was deficient. These results suggest that measures of structure and function are not necessarily complementary. Further, given its sensitivity for identifying deficits not evident with traditional imaging methods, these results highlight the utility of spTMS-fMRI, a method that directly and causally probes effective connectivity, as a tool for studying brain-based disorders.

HYPOTHESIS Among patients undergoing urgent surgery for left-sided diverticulitis, those admitted on weekends vs weekdays have higher rates of Hartmann procedure and adverse outcomes. DESIGN Analysis of data from the Nationwide Inpatient Sample between January 2002 and December 2008. Unadjusted and risk-adjusted generalized linear regression models were used. SETTING Academic research. PATIENTS Data on patients undergoing urgent surgery for acute diverticulitis. MAIN OUTCOME MEASURES Rates of Hartmann procedure vs primary anastomosis, complications, length of hospital stay, and total hospital charges. RESULTS In total, 31 832 patients were included; 7066 (22.2%) were admitted on weekends, and 24 766 (77.8%) were admitted on weekdays. The mean (SD) age of patients was 60.8 (15.3) years, and 16 830 (52.9%) were female. A Hartmann procedure was performed in 4580 patients (64.8%) admitted on weekends compared with 13 351 patients (53.9%) admitted on weekdays (risk-adjusted odds ratio [OR], 1.57; P &lt; .001). In risk-adjusted analyses, patients admitted on weekends had significantly higher risk for any postoperative complication (OR, 1.10; P = .005) and nonroutine hospital discharge (OR, 1.33; P &lt; .001) compared with patients admitted on weekdays, as well as a median length of hospital stay that was 0.5 days longer and median total hospital charges that were $3734 higher (P &lt; .001 for both). CONCLUSIONS Patients undergoing urgent surgery for left-sided diverticulitis who are admitted on a weekend have a higher risk for undergoing a Hartmann procedure and worse short-term outcomes compared with patients who are admitted on a weekday. Further research is warranted to investigate possible underlying mechanisms and to develop strategies for reducing this substantial weekend effect.

Abstract: Background: The Nissen and Toupet fundoplications are the most commonly used techniques for surgical treatment of gastroesophageal reflux disease. To date, no population-based trend analysis has been reported examining the choice of procedure and short-term outcomes. This study was designed to analyze trends in the use of Nissen versus Toupet fundoplications, and corresponding short-term outcomes during a 10-year period between 1995 and 2004. Methods: A trend analysis was performed of 873 patients (Toupet: 254 patients, Nissen: 619 patients) prospectively enrolled in the database of the Swiss Association for Laparoscopic and Thoracoscopic Surgery. Results: The frequency of the performed techniques remained stable during the observation period (p value for trend 0.206). The average postoperative and total length of hospital stay both significantly decreased during the 10-year period from 5.6 to 4.0 days and 6.8 to 4.8 days, respectively (both p values for trend <0.001). The average duration of surgery decreased significantly from 141 minutes to 121 minutes (p value for trend <0.001). There was a trend towards less complications in later years (2000–2004) compared to early years (1995–1999, p = 0.058). Conversion rates were significantly lower in later years compared with early years (p = 0.004). Conclusions: This is the first trend analysis in the literature reporting clinical outcomes of 873 prospectively enrolled patients undergoing Nissen and Toupet fundoplications during a 10-year period. The proportion of laparoscopic Nissen versus Toupet fundoplications remained stable over time, indicating that literature reports of the advantages of one procedure over the other had minimal influence on surgeons’ choice of technique. Length of hospital stay, duration of surgery, morbidity, and conversion rate decreased over time, reflecting the learning curve. Clearly, patient outcomes have much improved during the 10-year observation period.

Burnout is a pathologic reaction in response to long-term work-related stress. The aim of this study was 2-fold: first, to assess the prevalence and degree of burnout among surgical residents and surgeons in Switzerland and, second, to identify predictors of burnout in the surgical community. Four hundred five of 618 anonymous questionnaires (65.5%) were returned. Among respondents, 3.7% and 35.1% showed high and moderate degrees of burnout, respectively. Respondents with high and moderate degrees of burnout had higher summary scores of perceived stress (P &lt; .001). In multiple logistic regression analysis, the strongest predictors of burnout were poor interaction with nurses, disturbances due to telephone consultations, and high overall workload. To reduce burnout, new work models should be sought, in addition to decreasing work intensity and workload rather than restricting work hours alone.Arch Surg. 2010;145(10):1013-1016--

HYPOTHESIS The 50-hour workweek limitation for surgical residents in Switzerland has a major effect on surgical training, resident quality of life, and patient care. DESIGN Survey study. SETTING Residencies in Switzerland. PARTICIPANTS Surgical residents and surgical consultants. MAIN OUTCOME MEASURES An anonymous survey was conducted in Switzerland. Of 93 surgical departments contacted, 52 (55.9%) responded; of their 281 surgical residents and 337 surgical consultants, 405 (65.5%) returned a completed survey. RESULTS Residents and consultants indicated a negative effect of the 50-hour workweek limitation on surgical training (62.8% and 77.2%, respectively) and on quality of patient care (43.0% and 70.1%, respectively) (P &lt; .001 for both). Most residents and consultants reported that operative time (76.9% and 73.4%, respectively) and overall operating room experience (73.8% and 84.8%, respectively) were negatively affected by the work hour limitation. Only 8.1% of residents and 4.9% of consultants perceived the work hour limitation as beneficial to surgical training. Conversely, 58.4% of residents and 81.5% of consultants considered that residents' quality of life had improved (P &lt; .001). CONCLUSIONS Most surgical residents and surgical consultants perceive the work hour limitation as having a negative effect on surgical training and on the quality of patient care. Despite somewhat improved resident quality of life, the work hour limitation for surgical residencies in Switzerland appears to be a failure.Arch Surg. 2010;145(6):558-563--

HYPOTHESIS High-volume surgeons and hospitals are more likely to perform laparoscopic procedures than open procedures for diverticular disease as compared with low-volume surgeons and hospitals. DESIGN Real-world analysis. SETTING United States community hospitals. PATIENTS Patients with primary International Classification of Diseases, Ninth Revision diagnosis codes for diverticulosis or diverticulitis and International Classification of Diseases, Ninth Revision procedure codes for laparoscopic or open sigmoidectomy were selected from the 1992 to 2001 Nationwide Inpatient Samples commercially available US databases. MAIN OUTCOME MEASURES The outcome variable was the likelihood of performing laparoscopic vs open sigmoid resection. The primary predictor variable was the annual caseload of sigmoid resections per surgeon and hospital. RESULTS The study population included 55 949 patients who were predominantly white (70.5%) with a mean (SD) age of 60.7 (14.7) years. Unadjusted and risk-adjusted odds ratios of performing laparoscopic sigmoidectomy were significantly higher for high-volume surgeons and high-volume hospitals. In fact, high-volume surgeons were 8.80 times more likely to perform a laparoscopic sigmoid resection compared with low-volume surgeons. Similarly, in high-volume hospitals, patients were 3.02 times more likely to undergo a laparoscopic sigmoid resection compared with patients who underwent surgery in low-volume hospitals. These clinically relevant differences remained statistically significant in subset analyses stratified by age (&lt;65 vs ≥65 years) and time of surgery (elective vs nonelective). CONCLUSIONS The findings of the present investigation based on data from large US nationwide databases provide compelling evidence that high-volume surgeons and hospitals are significantly more likely to perform laparoscopic surgery for diverticular disease compared with low-volume surgeons and hospitals. Based on recent studies showing clear advantages of the laparoscopic technique over the open counterpart, our results should be considered by both patients and physicians.Arch Surg. 2007;142:253-259--

To determine national trends and putative racial and socioeconomic disparities in health care utilization in pediatric patients with pyogenic arthritis over a 13-year period.

Cancer immunotherapy is increasingly accepted as a treatment option for advanced stage disease. The identification of tumour-associated antigens in 1991 has prompted the development of antigen-specific immunotherapeutic strategies for a variety of cancers. Many of them result in some immunological responses in cancer patients; however, clinical results were not observed concomitantly with immunological responses; therefore, further improvements in the field of immunotherapy are urgently needed. Virosomes are lipidic envelopes devoid of genetic information, but which retain the antigenic profile and fusogenic properties from their viral origin. Virosomes are versatile antigen carriers and can be engineered to perform various tasks in cancer immunotherapy. Preclinical data have fostered the development of innovative clinical protocols. Hence, immunopotentiating reconstituted influenza virosomes will be assessed in breast and melanoma immunotherapy, and may contribute to the development of clinically effective cancer vaccines and ultimately improve patient outcomes. The objective of this review is to provide an overview of the potential clinical applications of virosomes as innovative and potentially effective reagents in active specific cancer immunotherapy.

Chorioamnionitis is associated with intense neutrophil infiltration of the decidua. We therefore determined whether chorioamnionitis enhances decidual interleukin-8 (IL-8) expression and examined cytokine-regulated decidual IL-8 expression. Decidua from chorioamnionitis-complicated pregnancies, but not term controls, displayed marked IL-8 immunohistochemical staining and a dense neutrophil infiltrate. Reverse transcriptase-polymerase chain reaction of microdissected decidual cells identified IL-8 mRNA, confirming decidual synthesis of IL-8. Confluent leukocyte-free term decidual cells were primed with 10−8mol/L estradiol (E2) or E2+ 10−7mol/L medroxyprogesterone acetate to mimic the steroidal milieu of pregnancy. Compared with cultures maintained in E2alone, E2+ medroxyprogesterone acetate neither significantly affected IL-8 levels nor altered the response to the cytokines. The addition of 1.0 ng/ml tumor necrosis factor-α (TNF-α) or interleukin-1 β (IL-1β) increased IL-8 secretion levels by 236.6 ± 51.4- and 1062.6 ± 254.3-fold, respectively (n= 8, mean ± SEM, P< 0.05), as measured by enzyme-linked immunosorbent assay. Concentration-response studies revealed that 0.01 ng/ml TNF-α and IL-1β elevated IL-8 output by 10- and 100-fold, respectively. Western blotting confirmed these results, and quantitative reverse transcriptase-polymerase chain reaction demonstrated parallel changes in mRNA levels. In conclusion, IL-8 is strongly expressed in term decidua during chorioamnionitis, and TNF-α and IL-1β enhance IL-8 expression in term decidual cells, suggesting that these cytokines are important regulators of chorioamnionitis-related decidual neutrophil infil-tration.

Longitudinal trends in epidemiology and utilization of total shoulder arthroplasty (TSA) have not been previously reported. We evaluated trends in the distribution of age, race, hospital volume and teaching status, outcomes, and indications for TSA during the last decade.TSA cases (n = 12,758) were extracted from the 1990–2000 Nationwide Inpatient Sample databases. TSA trends were obtained for 3 time periods: 1990–1993 (period I), 1994–1997 (period II), and 1998–2000 (period III).Between 1990 and 2000, there were minor increases in the rate of TSA in most age groups. Ninety‐three percent of the patients undergoing TSA in all 3 time periods were white. An increased proportion of patients were operated on in high volume hospitals in period III as compared with period I. Patients discharged to inpatient rehabilitation facilities after surgery had longer lengths of in‐hospital stays as compared with those discharged home. Osteoarthritis was diagnosed in an increasing proportion of patients undergoing TSA (56.6% in period I versus 70.9% in period III).There was a minor increase in the rate of TSA, and almost no change in use of TSA by nonwhites from 1990 through 2000. Efforts to understand and narrow this apparent underutilization of TSA among nonwhites are required. Further research should determine whether the observed shift of TSA to high volume centers improves surgical outcomes.

Summary: Background. Reports on long-term outcomes after endoscopic axillary lymph node dissection (ALND) of breast cancer patients are still lacking in the medical literature. The objective of this prospective study was to assess the oncological and functional outcomes in breast cancer patients after endoscopic ALND. Methods. Fifty-five breast cancer patients were prospectively enrolled, of whom 52 were available for follow-up with a median of 71.9thinspmonths (range 11–96). The following oncological and functional endpoints were evaluated during follow-up at several time points: occurrence of local, axillary and distant metastases, seroma or infection, shoulder mobility (range of motion), numbness, pain, presence of lymphoedema as well as restriction in activities of daily living. Results. In 52 patients endoscopic ALND of level I and II was successfully performed. Two port-site metastases (2/52, 4%) occurred, one of which in a patient with negative axillary lymph nodes. The same patient suffered from the only axillary recurrence (1/52, 2%). Three patients (3/52, 6%) developed lymphoedema. No other functional adverse events (shoulder mobility, pain, numbness, hypertrophic scar) were noticed at the end of the observation period. Conclusion. The present investigation with long-term follow-up after endoscopic ALND – the first one in the literature – reveals minor morbidity, good functional and cosmetic results. In contrary to conventional surgery, the endoscopic procedure is associated with the occurrence of port-site metastases, not seen in the open approach. Axillary recurrences do not appear more frequently when compared with results after conventional ALND. In the meantime the less invasive sentinel lymph node (SLN) biopsy is the established standard technique in evaluating the axillary lymph node status.

Purpose: Regional anesthesia can be the technique of choice for selected ambulatory surgery procedures, but in spite of its benefits, it has an inherent failure rate even in experienced hands. We examine the efficacy and factors associated with failure of ambulatory regional anesthesia techniques. Methods: This study included 9,342 blocks performed on 7,160 patients at the Duke University Ambulatory Surgery Center. Blocks were classified as interscalene, supraclavicular, axillary, lumbar plexus, femoral, sciatic, ankle, paravertebral, spinal, and other (frequency less than 100). A block was considered surgical if a single attempt at placing the block resulted in a complete sensory, motor, and sympathetic nerve block. Multiple logistic regression analyses were used to assess the risk-adjusted association between patient characteristics and block failure. Results: Paravertebral blocks and those considered in the “other” category had significantly higher failure rates (P&lt; 0.001 ), while spinal and lumbar plexus blocks had lower than average rates of failure (P&lt; 0.001 andP= 0.03, respectively). In multiple logistic regression analyses excluding paravertebral blocks, body mass index (BMI) scores greater than 25 (Pvalues: BMI 25–29: &lt; 0.001; BMI 30–34:P&lt; 0.001; BMI 35:P&lt; 0.001) and ASA physical status IV (P&lt; 0.001 ) were significantly associated with higher block failure rates. Conclusion: High BMI and ASA IV are independent risk factors for block failure in ambulatory surgery patients.

Extracellular matrix (ECM) proteins synthesized by human placental mesenchymal cells (PMCs) provide structural support for the villus. Aberrant expression of ECM proteins by PMCs has been associated with intrauterine growth restriction (IUGR). To provide insight into the mechanisms of ECM protein regulation in the stroma of the placental villus, in the current study, we examined the interaction of glucocorticoid (GC) and transforming growth factor-β (TGFβ) in the modulation of ECM proteins in cultures of PMCs isolated from human term placentas. Initial results obtained by ELISA showed that combined treatment with dexamethasone (DEX) and TGFβ enhanced oncofetal fibronectin (FFN) protein levels in serum-free culture medium severalfold in a dose-dependent manner. Northern blotting and real-time polymerase chain reaction (PCR) analyses revealed a similar enhancement in levels of FN mRNA in cells treated with TGFβ and DEX. Real-time PCR results also revealed that DEX and TGFβ enhanced collagen (Col) I and Col IV expression, but did not affect levels of Col III or laminin, indicative of selective stimulation of ECM proteins. Hypoxic treatment moderately enhanced FFN levels in control cells but not in those treated with DEX and TGFβ. In contrast with the results obtained with PMCs, we noted that DEX treatment suppressed FFN levels in untreated and TGFβ-treated cytotrophoblasts, suggesting that GC and TGFβ modulate FFN expression in placenta in a cell-type-specific manner. We conclude that GC and TGFβ are key regulators of ECM protein synthesis in PMCs, suggesting a role in modulating placental architecture in uncomplicated pregnancies and those associated with aberrant ECM protein expression.

We performed a phase I/II clinical trial in metastatic melanoma patients with an ultraviolet (UV)-inactivated nonreplicating recombinant vaccinia virus enabling the expression, from a single construct, of endoplasmic reticulum-targeted HLA-A0201-restricted Melan-A/MART-127-35, gp100280-288, and tyrosinase1-9 epitopes, together with CD80 and CD86 costimulatory proteins. Corresponding soluble peptides were used to boost responses and granulocyte-macrophage colony-stimulating factor was used as systemic adjuvant. Safety and immunogenicity, as monitored with in vitro-restimulated peripheral blood mononuclear cells by cytotoxic T lymphocyte precursor (CTLp) frequency analysis and tetramer staining, were specifically addressed. Of 20 patients entering the protocol, 2 had to withdraw because of rapidly progressing disease. Immune responses were evaluated in 18 patients (stage III, n = 5; stage IV, n = 13) and increases in specific CTLp frequencies were observed in 15. In 16 patients responsiveness against all 3 antigens could be analyzed: 7 (43%), including all stage III cases, showed evidence of induction of CTLs specific for the three epitopes, and 2 (12%) and 4 (25%), respectively, showed reactivity against two or one tumor-associated antigen. In three stage IV patients no specific CTL reactivity could be induced. Increases in CTLp frequency were detected mostly after viral vaccine injections. However, in a majority of patients final CTLp levels were comparable to initial levels. Tetramer characterization of Melan-A/MART-127-35-specific CTLs during the protocol also suggested preferential expansion after recombinant virus administration. Vector-specific humoral responses, frequently undetectable in stage IV patients, did not appear to prevent tumor-associated antigen-specific CTL induction. Aside from a single occurrence of transient grade 3 leukopenia, no major clinical toxicity was reported. Seventeen of 18 patients completed the 3-month trial (one patient died before the last delayed-type hypersensitivity test). Three displayed regression of individual metastases, seven had stable disease, and progressive disease was observed in seven patients. This is the first report on the administration of a UV-inactivated recombinant vaccinia virus coexpressing five transgenes in cancer patients. The results described here, in terms of safety and immunogenicity, support the use of this reagent in active specific immunotherapy.

Plasminogen activator inhibitor (PAI)-1 plays a key role in the regulation of fibrinolysis and cellular invasion by virtue of suppression of plasminogen activator function. Excessive production of placental PAI-1 has been associated with aberrant periplacental fibrin deposition in pregnancies complicated by pre-eclampsia (PE) and intrauterine growth restriction (IUGR). In the current study we used HTR-8/SVneo cells and primary cultures of cytotrophoblasts as models for study of PAI-1 regulation by transforming growth factor (TGF)-β, and dexamethasone (DEX), a synthetic glucocorticoid (GC). ELISA and Northern blotting assays revealed that DEX treatment significantly enhanced TGF-β effects on PAI-1 protein and mRNA expression in HTR-8/SVneo cells and cytotrophoblasts. These effects were GC-specific in that DEX and cortisol, but not estradiol, progesterone or testosterone, augmented PAI-1 levels in TGF-β-treated cells. Conversely, DEX and TGF-β treatment suppressed PAI-2 levels in HTR-8/SVneo cells and did not affect PAI-2 levels in cytotrophoblasts. PAI-1 promoter assays revealed that TGF-β, but not DEX, enhanced PAI-1 expression in HTR-8/SVneo cells through a transcriptional mechanism. These results suggest that GC and TGF-β may alter fibrinolytic and invasive properties of trophoblasts through their effects on PAI-1 expression.