Search

Transformative results of adeno-associated virus (AAV) gene therapy in patients with spinal muscular atrophy and Leber’s congenital amaurosis led to approval of the first two AAV products in the United States to treat these diseases. These extraordinary results led to a dramatic increase in the number and type of AAV gene-therapy programs. However, the field lacks non-invasive means to assess levels and duration of therapeutic protein function in patients. Here, we describe a new magnetic resonance imaging (MRI) technology for real-time reporting of gene-therapy products in the living animal in the form of an MRI probe that is activated in the presence of therapeutic protein expression. For the first time, we show reliable tracking of enzyme expression after a now in-human clinical trial AAV gene therapy (ClinicalTrials.gov: NTC03952637) encoding lysosomal acid beta-galactosidase (βgal) using a self-immolative βgal-responsive MRI probe. MRI enhancement in AAV-treated enzyme-deficient mice (GLB-1−/−) correlates with βgal activity in central nervous system and peripheral organs after intracranial or intravenous AAV gene therapy, respectively. With >1,800 gene therapies in phase I/II clinical trials (ClinicalTrials.gov), development of a non-invasive method to track gene expression over time in patients is crucial to the future of the gene-therapy field., Graphical abstract, We describe a bio-responsive contrast agent that informs on the biodistribution and durability of gene expression in the lysosomal storage disease, GM1 gangliosidosis. We illustrate the sensitivity of this contrast agent to detect β-galactosidase enzyme in the central nervous system and periphery of an AAV-GLB-1-treated mouse model of GM1.

The GM2 gangliosidoses, Tay-Sachs disease (TSD) and Sandhoff disease (SD), are fatal lysosomal storage disorders caused by mutations in the HEXA and HEXB genes, respectively. These mutations cause dysfunction of the lysosomal enzyme β-N-acetylhexosaminidase A (HexA) and accumulation of GM2 ganglioside (GM2) with ensuing neurodegeneration, and death by 5 years of age. Until recently, the most successful therapy was achieved by intracranial co-delivery of monocistronic adeno-associated viral (AAV) vectors encoding Hex alpha and beta-subunits in animal models of SD. The blood-brain barrier crossing properties of AAV9 enables systemic gene therapy; however, the requirement of co-delivery of two monocistronic AAV vectors to overexpress the heterodimeric HexA protein has prevented the use of this approach. To address this need, we developed multiple AAV constructs encoding simultaneously HEXA and HEXB using AAV9 and AAV-PHP.B and tested their therapeutic efficacy in 4- to 6-week-old SD mice after systemic administration. Survival and biochemical outcomes revealed superiority of the AAV vector design using a bidirectional CBA promoter with equivalent dose-dependent outcomes for both capsids. AAV-treated mice performed normally in tests of motor function, CNS GM2 ganglioside levels were significantly reduced, and survival increased by >4-fold with some animals surviving past 2 years of age.

Eccrine porocarcinoma is a rare malignant tumor that develops in the eccrine glands, appearing as a primary tumor, or by malignant transformation of an eccrine poroma. It is a carcinoma with high metastatic and recurrent potential; it has the same incidence in both sexes, and mainly affects the elderly. Its diagnosis, rather than clinical, is histological, and due to the rarity of the disease, it is a pathological challenge. There are no standardized treatment guidelines for porocarcinoma, but surgical resection with tumor-free margins is considered the basis of treatment, in addition to sentinel node biopsy under risk factors and individualization of each patient. For the metastatic form, chemotherapy and radiotherapy are the treatment of choice. Herein, we present the case of a man with eccrine porocarcinoma with extensive zosteriform skin metastasis and lymph node involvement, treated with chemotherapy and concomitant radiotherapy.

Disparities persist in breast cancer outcomes between Latina survivors and non-Hispanic Whites. Identifying methods to ensure that Latinas participate in and benefit from translational behavioral medicine research is important to reduce disparities. We developed a “Social Network Research Registry” to enhance Latina survivors’ engagement in research and explored the social networks and research/cancer organization participation in this population. We initially recruited 30 Latina breast cancer survivors (“seeds”) from community organizations and identified other survivors through snowball sampling. Guided by Social Network Analysis, we assessed the structural (e.g., size) and functional (e.g., social support) characteristics of the network, willingness to join the registry, prior research participation, involvement in cancer organizations, and interest in different types of research and roles in research. The resulting network size was 98, including 53 women who enrolled in the study and 45 who were listed in the network but did not enroll. All enrolled participants (N = 53) agreed to be part of the registry. We identified 15 participants who occupied strategic positions as hubs and/or bridges. Women who were currently involved in cancer organizations were more likely to have participated in research (70.3% vs. 18.8%); χ(2) (1, 53) = 11.97, p = .001. Most were interested in surveys/interviews (98%), behavioral interventions (96%), and becoming health promoters (79%). The Social Network Research Registry is an acceptable and feasible strategy to engage underrepresented Latina survivors in research. Social network analysis can be useful to identify members who occupy key positions to enhance recruitment and translational efforts.

Ravulizumab is a long-acting C5 inhibitor engineered from eculizumab with increased elimination half-life, allowing an extended dosing interval from two to eight weeks. Here we evaluate the efficacy and safety of ravulizumab in adults with atypical hemolytic uremic syndrome presenting with thrombotic microangiopathy. In this global, phase 3, single arm study in complement inhibitor-naive adults (18 years and older) who fulfilled diagnostic criteria for atypical hemolytic uremic syndrome, enrolled patients received ravulizumab through a 26-week initial evaluation period. The primary endpoint was complete thrombotic microangiopathy response defined as normalization of platelet count and lactate dehydrogenase and 25% or more improvement in serum creatinine. Secondary endpoints included changes in hematologic variables and renal function. Safety was also evaluated. Ravulizumab treatment resulted in an immediate, complete, and sustained C5 inhibition in all patients. Complete thrombotic microangiopathy response was achieved in 53.6% of patients. Normalization of platelet count, lactate dehydrogenase and 25% or more improvement in serum creatinine was achieved in 83.9%, 76.8% and 58.9% of patients, respectively. Improvement in estimated glomerular filtration rate by one or more stage was achieved in 68.1% of patients by day 183. No unexpected adverse events were reported across a safety analysis set of 58 patients. Four deaths occurred (three within one month of study initiation, including one in a patient excluded based on eligibility criteria after the first dose) with none considered treatment-related by the study investigator. Thus, treatment with ravulizumab once every eight weeks resulted in rapidly improved hematologic and renal endpoints with no unexpected adverse events in adults with atypical hemolytic uremic syndrome.

The authors regret that 2 investigators were not included in the 311 Study Group. Members of the 311 Study Group are listed in the Appendix. The authors would like to apologize for any inconvenience caused.

The “westernization” of global eating and lifestyle habits is associated with the growing rate of chronic diseases, mainly cardiovascular diseases, cancer, type 2 diabetes mellitus, and respiratory diseases. The primary prevention approach is to make nutritional and behavioral changes, however, there is another important determinant of our health that only recently has been considered and is the presence of beneficial microorganisms and their products in our gastrointestinal tract. Microorganisms living in our body can alter the fate of food, drugs, hormones, and xenobiotics, and recent studies point to the use of microorganisms that can counteract the harmful effects of certain compounds introduced or produced endogenously in our body. This review considers the effects of the western lifestyle on adiposity, glucose metabolism, oxidative markers and inflammation profile, emphasizes on the studies that have investigated bacterial strains and products of their metabolism that are beneficial under this lifestyle, and examines the screening strategies that recent studies are using to select the most promising probiotic isolates. In addition, we consider the relevance of studying the microbiota of metabolically healthy people under a western lifestyle for the understanding of the key components that delay the development of chronic diseases.

Secreted frizzled related protein (sFRP)-2, a Wnt antagonist, was strongly expressed by both stromal and tumor cells of ameloblastoma. The aim of this study is to evaluate whether sFRP-2 secreted from tumor cells have any direct role in suppressed bone formation or not. A pre-osteoblastic cell line, KUSA/A1 cells, cultured in conditioned medium of an ameloblastoma-derived cell line (AM-1CM) was used in the study. Alkaline phosphatase (ALP) activity, alizarin red staining, mineral quantification and MTS assay was performed. Wnt-canonical pathway is a major pathway for osteoblasts. Antagonists of this pathway, sFRP-1, 2 and 3, were detected by immunohistochemistry and western blot analysis. KUSA/A1 cells cultured in AM-1CM showed high cell proliferation, low ALP activity without mineralized matrix deposition. sFRP-2 was strongly expressed in ameloblastoma tissue and AM-1 cells. After sFRP-2 depletion, the cells showed diffuse mineralization. In this study, it was confirmed that ameloblastoma cells have a major role in decreased bone formation by secreting sFRP-2 in cell culture model. Though, sFRP-2 has great effect on tumor progression, inhibition of sFRP-2's anti-bone formation activity and cell proliferative activity may reduce the invasive property of ameloblastoma and possibility of recurrence rate.

Loss of heterozygosity (LOH) in the ING family members has been shown in head and neck squamous cell carcinoma (HNSCC) except for ING2. Like all the other members of ING family, ING2, which is located at chromosome 4q35.1, is a promising tumor suppressor gene (TSG). In this study, we performed LOH analysis of ING2 in HNSCC and compared it with clinicopathological variables.We performed LOH analysis in DNAs from 80 paired of normal and HNSCC tissues, using a specifically designed microsatellite marker on chromosome 4q35.1, which detects allelic loss of ING2. TP53 mutation analysis and its relationship with ING2 chromosomal deletion were also performed in available 68 of the samples. The correlation between LOH status and clinicopathological characteristics was evaluated by using statistical methods. The overall survival (OS) and disease free survival (DFS) were also determined.LOH was detected in 54.6% (30/55) of the informative samples. Statistical significance was obtained between LOH and tumor (T) stage (P = 0.02), application of radiotherapy and chemotherapy. Positive node status (N) appeared to be the only independent prognostic factor for both OS (P = 0.031) and DFS (P = 0.044).Our study showed allelic loss of 4q35.1 in HNSCC. The high percentage of LOH suggests ING2 as a candidate TSG in HNSCC. High LOH frequency was statistically associated with advanced T stage, suggesting that ING2 LOH might occur in late stages during HNSCC progression.

The Brief Symptom Inventory-18 (BSI-18) is an 18-item symptom checklist used as a brief distress screening in cancer and other medical patients. This study evaluated the validity of the BSI-18 in a sample of 221 adult survivors of childhood cancers ages 18-55 (median = 26). Validity of the BSI-18 was compared to the Symptom Checklist-90-Revised (SCL-90-R). Results indicated the BSI-18 scales had acceptable internal consistency (alpha >0.80) and were highly correlated with the corresponding SCL-90-R subscales (correlations from 0.88 to 0.94). When subjects were classified as case positive (significantly distressed) using the BSI-18 manual case-rule, classification agreement with the SCL-90-R was poor as evidenced by low sensitivity (41.78%). An alternative BSI-18 case-rule previously developed for cancer patients using the General Severity Index (GSI; GSI t-score >or=57) demonstrated better sensitivity (83.54%). ROC analysis indicated the BSI-18 had strong diagnostic utility relative to the SCL-90-R (AUC = 0.98) and several possible GSI cut-off scores were evaluated. The optimal cut-of score was a t-score >or=50 which had a sensitivity of 97.47% and a specificity of 85.21%. Results support use of the BSI-18 with adult survivors of childhood cancer but indicate an alternative case-rule must be used.

Bone and dentin resemble each other in composition and mechanism of formation and mineralization. Preliminary analysis of osteoblastic and odontoblastic cells before using them in tissue engineering is mandatory. Thus, in order to evaluate the process of calcification, we evaluated osteoblastic-like (KUSA/A1cells) and odontoblastic-like (MDPC-23 cells) cells seeded in cell culture and in intraperitoneal diffusion chamber. Our results indicated that KUSA/A1 cells differentiated into osteoblasts-like cells and induced bone tissue inside the chamber. Whereas, MDPC-23 cells showed odontoblastic phenotype without ability to induce dentin formation, suggesting that MDPC-23 cells are special cells, which lost the capacity to induce mineralized dentin matrix after long period of time. This study showed the significance of basic information of calcification process by osteoblastic-like and odontoblastic-like cells before using them in tissue engineering.

BACKGROUND: To validate a new practical Sepsis Severity Score for patients with complicated intra-abdominal infections (cIAIs) including the clinical conditions at the admission (severe sepsis/septic shock), the origin of the cIAIs, the delay in source control, the setting of acquisition and any risk factors such as age and immunosuppression. ----- METHODS: The WISS study (WSES cIAIs Score Study) is a multicenter observational study underwent in 132 medical institutions worldwide during a four-month study period (October 2014-February 2015). Four thousand five hundred thirty-three patients with a mean age of 51.2 years (range 18-99) were enrolled in the WISS study. ----- RESULTS: Univariate analysis has shown that all factors that were previously included in the WSES Sepsis Severity Score were highly statistically significant between those who died and those who survived (p

Cardiovascular disease is a serious public health problem worldwide, occupying the first place among the causes of morbidity and mortality and also with a high potential for disability. According to the World Health Organization (WHO), cardiovascular disease deaths represent 30% of all deaths worldwide. The oil and gas industry is not an exception to this problem, with statistics for an oilfield services company over the last 10 years showing that more than 150 employees died of cardiovascular disease; 95% from heart attack and 5% from stroke. In Colombia it was found in 2012 that a high percentage of the working population of the oilfield services company had one or more risk factors for coronary heart disease: 48.5% for overweight and obesity, 32% for lipid disorders, and 81% for physical inactivity (sedentarism). In consideration that many of the risk factors for cardiovascular disease are modifiable, such as improving one’s lifestyle concerning diet and exercise, a program named Ergoday was structured and implemented to focus on providing an individual diagnosis, intervention, and monitoring of the modifiable risk factors. With an emphasis on weight control, muscle strength, and flexibility in addition to the control of cardiovascular risk factors, the aim of the Ergoday program is to control both cardiovascular risk and musculoskeletal disorders, which were also prevalent in the Colombia workforce. Participants in the program become aware of the risk factors for cardiovascular disease and musculoskeletal disorders (MSD) and the need for physical fitness to improve their quality of life. To achieve long-term success, the intervention includes specific recommendations to reduce or eliminate the risk factors and regular monitoring to assess compliance and impact of the recommendations given.

Four postmortem cases are reported in which the selective serotonin re-uptake inhibitor fluvoxamine was identified. Fluvoxamine was detectable using a standard alkaline drug screen, chromatographed well using a HP-1 column, and did not require derivitization for quantitation. Two of the cases reported were drug intoxications; fluvoxamine was only an incidental finding in the other 2 cases. Central and peripheral blood values are reported, as well as antemortem blood, bile, vitreous fluid, and urine values. No solid organs were obtained in any of the cases. Quantitations were performed using both an analytical standard and a fluvoxamine tablet for the preparation of calibrators. A comparison of quantitative values was made to evaluate the feasibility of using a tablet as the drug source for the preparation of calibrators when a pure reference material is unavailable. Postmortem peripheral blood concentrations ranged from approximately 0.5 mg/L in a case of suicidal shooting to approximately 6 mg/L in a case of drug overdose. Evidence of postmortem redistribution was noted in the only case for which both central and peripheral blood were obtained. Quantitations using an extracted drug tablet for the preparation of calibrators correlated well with quantitations using a pure reference material.

INTRODUCTION Research has consistently shown posttraumatic stress disorder (PTSD) to be associated with impairments in functioning across a number of psychosocial domains [1]. Specifically, PTSD is associated with impairments in occupational and academic functioning [2-6], marital and family functioning [5,7-8], parenting [9-10], and friendships and socializing [11]. Additional studies have shown associations between PTSD and objective indicators of quality of life (QOL) such as homelessness and unemployment [12-13]. Such impairments are common among populations at high risk for PTSD, such as military personnel deployed to combat [3,5-6,8-14]. Research suggests that these impairments are currently affecting many Veterans of the wars in Iraq and Afghanistan (Operations Iraqi Freedom and Enduring Freedom) and are therefore important to identify and treat [15]. In 2010, the Department of Veterans Affairs (VA) and the Department of Defense (DOD) jointly published the revised VA/DOD Clinical Practice Guideline for Management of Posttraumatic Stress [16]. This clinical practice guideline (CPG) provides evidence-based recommendations to clinicians for diagnosing and treating a spectrum of stress-related disorders including combat and operational stress reaction (COSR), acute stress reaction, acute stress disorder (ASD), acute PTSD, and chronic PTSD among servicemembers and Veterans. The VA/DOD CPG recommends a comprehensive assessment of all relevant domains of functioning. It stresses the importance of a thorough assessment of functional impairment for several reasons: (1) to identify individuals who may be at risk for endangering themselves or others during or after deployment as well as after military discharge, (2) to promote accurate diagnosis, (3) to guide treatment planning by clarifying the domains in which the individual is experiencing impairment, and (4) to monitor changes in functioning during and after treatment. This article is meant to complement the CPG recommendations for assessing stress disorder-related functional impairment and will provide information that may further guide clinicians in their assessment efforts. Although the VA/DOD CPG addresses several stress-related disorders, this article will focus mainly on the relation between PTSD and psychosocial functioning because the vast majority of prior research has examined this association. The information provided is based on these empirical findings from the extant literature as well as recent findings from an ongoing study of functional impairment among male and female Veterans. Specifically, we begin our article with a review of some of the recent empirical literature on the association between PTSD and impaired functioning across various psychosocial domains. We then provide specific recommendations on how to perform a comprehensive multimethod assessment of functional impairment and introduce a promising new assessment instrument. The article concludes with insights into several important issues related to assessing functional impairment that we encourage all clinicians to keep in mind when assessing functional impairment among Veterans and Active Duty servicemembers. METHODS We searched the U.S. National Library of Medicine's PubMed, PsycINFO, and PsycARTICLES databases for articles related to PTSD and functioning. We identified studies by searching the databases for references with the phrases "posttraumatic stress disorder" or "PTSD" (n = 10,109 English-language articles) or "functioning" in the title or abstract (n = 83).We reviewed the abstracts for the resulting articles to identify those relevant to our topic, and we also reviewed the references for the most relevant articles to identify additional studies of interest. Because we were unable to provide an exhaustive literature review in this article, we emphasized studies published since 2008 but also included a few earlier articles that were of particular relevance. …

Ameloblastoma is the most frequently encountered odontogenic tumor, characterized by a locally invasive behavior, frequent recurrences, and, although rare, metastatic capacity. Loss or inactivation of tumor suppressor genes (TSGs) allows cells to acquire neoplastic growth. The ING family proteins are tumor suppressors that physically and functionally interact with p53 to perform important roles in apoptosis, DNA repair, cell cycle regulation, and senescence. TP53 genetic alterations were reported to infrequently occur in ameloblastoma. Considering that other TSGs related to TP53 could be altered in this tumor, we focused our study on the ING family genes. We analyzed the loss of heterozygosity (LOH) status of the ING family (ING1-ING5) chromosomal loci in a group of ameloblastomas by microsatellite analysis, and correlated the ING LOH status with clinicopathological characteristics. By using specific microsatellite markers, high frequency of LOH was found at the loci of each ING gene family member (33.3-72.2%). A significant relationship was shown between LOH of D2S 140 (ING5 locus) and solid tumor type (p = 0.02). LOH of ING3MS (ING3 locus) was also high in solid type tumors, showing a near significant association. In addition, a notable tendency toward higher LOH for half of the markers was observed in recurrent cases. LOH of ING family genes appears as a common genetic alteration in solid ameloblastoma. The current study provides interesting novel information regarding the potential prognostic significance of the allelic loss of the ING gene family loci in ameloblastoma tumorigenesis.

Fanconi anemia is a rare autosomal recessive or X-linked genetic disease characterized by progressive bone marrow failure, various developmental anomalies, and cancer predisposition. Fanconi anemia complementation group A (FANCA) gene is one of the 15 disease-causing genes and has been found to be mutated in ∼60% of Fanconi anemia patients. Except for its constituent role in the ubiquitin ligase of the Fanconi anemia core complex, no other biochemical activity has been identified thus far. Using purified protein, we observed that human FANCA has intrinsic affinity to nucleic acids. FANCA binds to both single-stranded and double-stranded DNA; however, its affinity to single-stranded DNA is dramatically higher. FANCA also binds to RNA with an intriguingly higher affinity than its DNA counterpart. A patient-derived FANCA truncation mutant, Q772X, has diminished affinity to both DNA and RNA. On the contrary, the complementing C-terminal fragment of Q772X, C772-1455, retains the differentiated nucleic acids binding activity (RNA>ssDNA>dsDNA), indicating that the nucleic acids binding domain of FANCA is primarily located at its C-terminus. Using DNA and RNA ladders, we determined that the minimum numbers of nucleotides required for FANCA recognition are ∼30 for both DNA and RNA. By testing the affinity between FANCA and a variety of DNA structures, we found that a 5’ flap or tail on DNA facilitates its interaction with FANCA. In addition, we observed that FANCA greatly stimulates the endonuclease activity of flap endonuclease 1 (FEN1), which processes 5’ flap structures. Our study reported that purified human FANCA binds to nucleic acids with strong preference for single-stranded forms. Our finding suggests that FANCA FANCA may participate in maintenance of replication forks by regulating removal of Okazaki fragment during maturation of lagging strand replication. Citation Format: Liangyue Qian, Fenghua Yuan, Paola Rodriguez-Tello, Xinliang Zhao, Limin Song, Gennaro D'Urso, Chaitanya Jain, Yanbin Zhang. FANCA binds to nucleic acids and stimulates endonuclease activity of FEN1. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1286. doi:10.1158/1538-7445.AM2013-1286