Multidisciplinary consensus statement on the clinical management of patients with stage III non-small cell lung cancer.

Stage III non-small cell lung cancer (NSCLC) is a very heterogeneous disease that encompasses patients with resected, potentially resectable and unresectable tumours. To improve the prognostic capacity of the TNM classification, it has been agreed to divide stage III into sub-stages IIIA, IIIB and IIIC that have very different 5-year survival rates (36, 26 and 13%, respectively). Currently, it is considered that both staging and optimal treatment of stage III NSCLC requires the joint work of a multidisciplinary team of expert physicians within the tumour committee. To improve the care of patients with stage III NSCLC, different scientific societies involved in the diagnosis and treatment of this disease have agreed to issue a series of recommendations that can contribute to homogenise the management of this disease, and ultimately to improve patient care.

Sacral neuromodulation for fecal incontinence in Latin America: initial results of a multicenter study.

BACKGROUND: Sacral neuromodulation (SNM) is a widely used therapeutic option for fecal incontinence (FI). Larger series are mainly from Western countries, while few reports address the results of SNM in less developed or less wealthy countries. The aim of the present study was to evaluate the efficacy of SNM in patients with FI in Latin America. METHODS: A retrospective study was conducted on patients with FI who had SNM between 2009 and 2016 at 15 specialized colorectal surgery centers in Latin America. Main outcomes measures were functional outcomes, postoperative complications, requirement of revisional surgery, and requirement of device removal. All patients had failed conservative management and had clinical assessment including recording of the validated Cleveland Clinic Florida Fecal Incontinence Score (CCF-FIS) and, when available, anal manometry and endoanal ultrasound. Patients were followed up for a median of 36.7 (1-84) months. RESULTS: One hundred and thirty-one patients [119 females, median age of 62.2 (range 19-87) years] were included. The most common etiology of FI was obstetric injury (n = 60; 45.8%). After successful test lead implantation, the stimulator was permanently placed in 129 patients (98.5%). One patient failed to respond in the test phase and one patient did not proceed to permanent implantation for insurance reasons. Nineteen patients (14.7%) had 19 complications including infection (n = 5, 3.8%), persistent implant site pain (n = 5, 3.8%), generator/lead dislodgment (n = 5, 3.8%), malfunctioning device (n = 3, 2.3%), and hematoma (n = 1, 0.7%). Reimplantation after the first and second stages was necessary in 2 (1.5%) and 3 patients (2.3%), respectively. The device removal rate was 2.2%. At a median follow-up of 36.7 (range 1-84) months, the CCF-FIS significantly improved from a preoperative baseline of 15.9 ± 2.98 to 5.2 ± 3.92 (95%CI: 15.46 vs 4.43; p < 0.0001). Overall, 90% of patients rated their improvement as "significant". CONCLUSIONS: Sacral nerve stimulation for FI is safe and efficient, even in less wealthy or less developed countries.

Dendritic Immunotherapy Improvement for an Optimal Control Murine Model.

Therapeutic protocols in immunotherapy are usually proposed following the intuition and experience of the therapist. In order to deduce such protocols mathematical modeling, optimal control and simulations are used instead of the therapist's experience. Clinical efficacy of dendritic cell (DC) vaccines to cancer treatment is still unclear, since dendritic cells face several obstacles in the host environment, such as immunosuppression and poor transference to the lymph nodes reducing the vaccine effect. In view of that, we have created a mathematical murine model to measure the effects of dendritic cell injections admitting such obstacles. In addition, the model considers a therapy given by bolus injections of small duration as opposed to a continual dose. Doses timing defines the therapeutic protocols, which in turn are improved to minimize the tumor mass by an optimal control algorithm. We intend to supplement therapist's experience and intuition in the protocol's implementation. Experimental results made on mice infected with melanoma with and without therapy agree with the model. It is shown that the dendritic cells' percentage that manages to reach the lymph nodes has a crucial impact on the therapy outcome. This suggests that efforts in finding better methods to deliver DC vaccines should be pursued.

HMG20A is required for SNAI1-mediated epithelial to mesenchymal transition.

HMG20A is a high mobility group (HMG) domain containing protein homologous to HMG20B, a core subunit of the Lys-specific demethylase 1/REST co-repressor 1 (LSD1-CoREST) histone demethylase complex. Here, we show that HMG20A can replace HMG20B and, therefore, they are mutually exclusive subunits of the complex. Both proteins interact through a coiled-coil domain with BHC80, another subunit of the LSD1-CoREST complex. To investigate the functional differences between the two proteins, we performed transcriptomic analysis of HMG20A- and HMG20B-depleted cells. Analysis of the misregulated genes in HMG20A-knockdown cells evidenced a high proportion of genes related to the epithelial-to-mesenchymal transition (EMT) process. EMT occurs during embryonic development or during the course of malignant cancer progression and consists in the dynamic and reversible transitions between epithelial and mesenchymal phenotypes. We show that HMG20A together with LSD1 are required for SNAI1-dependent repression of epithelial genes and for (transforming growth factor ß) TGF-ß-triggered EMT. Importantly, HMG20A-depleted cells displayed reduced binding of LSD1 to epithelial gene promoters and increased methylation of lysine 4 of histone H3, suggesting a role of HMG20A in recruiting or in stabilizing the complex at the chromatin. SNAI1 and the TGF-ß-related transcription factor SMAD4 were found to be associated with the LSD1-CoREST complex containing HMG20A. Furthermore, we show that HMG20A-depleted cells displayed reduced motility and invasion activity. Finally, we show that expression of HMG20A correlates positively with mesenchymal markers and negatively with epithelial markers in human tumor samples. Taken together, our data demonstrate that HMG20A is essential for the mesenchymal phenotype.

Cytoplasmic interaction of the tumour suppressor protein hSNF5 with dynamin-2 controls endocytosis.

Human SNF5 (hSNF5; INI1, SMARCB1 or BAF47) is a component of the human SWI/SNF chromatin remodelling complex and a tumour suppressor mutated in rhabdoid tumours. It also associates with the integrase of the human immunodeficiency virus (HIV)-1. We show by fluorescence loss in photobleaching that hSNF5 is constantly shuttling between the nucleus and the cytoplasm, raising the question of what the role of hSNF5 is in the cytoplasm. Here, we demonstrate that hSNF5 directly interacts with the GTPase dynamin-2 (DNM2) in the cytoplasm. DNM2 is a large GTPase involved in endocytosis and vesicle dynamics, which has been related to HIV-1 internalization. We show that hSNF5 colocalizes with DNM2 in endocytic vesicles. Depletion of hSNF5, but not of other components of the SWI/SNF complex, destabilizes DNM2 and impairs DNM2-dependent endocytosis. Furthermore, we show that hSNF5 inhibits assembly-stimulated DNM2 GTPase activity but not basal GTPase activity in vitro. Altogether, these results indicate that hSNF5 affects both the stability and the activity of DNM2, uncovering an unexpected role of hSNF5 in modulating endocytosis, and open new perspectives in understanding the role of hSNF5 in tumour genesis.

The chromatin remodeling factor CHD8 interacts with elongating RNA polymerase II and controls expression of the cyclin E2 gene.

CHD8 is a chromatin remodeling ATPase of the SNF2 family. We found that depletion of CHD8 impairs cell proliferation. In order to identify CHD8 target genes, we performed a transcriptomic analysis of CHD8-depleted cells, finding out that CHD8 controls the expression of cyclin E2 (CCNE2) and thymidylate synthetase (TYMS), two genes expressed in the G1/S transition of the cell cycle. CHD8 was also able to co-activate the CCNE2 promoter in transient transfection experiments. Chromatin immunoprecipitation experiments demonstrated that CHD8 binds directly to the 5' region of both CCNE2 and TYMS genes. Interestingly, both RNA polymerase II (RNAPII) and CHD8 bind constitutively to the 5' promoter-proximal region of CCNE2, regardless of the cell-cycle phase and, therefore, of the expression of CCNE2. The tandem chromodomains of CHD8 bind in vitro specifically to histone H3 di-methylated at lysine 4. However, CHD8 depletion does not affect the methylation levels of this residue. We also show that CHD8 associates with the elongating form of RNAPII, which is phosphorylated in its carboxy-terminal domain (CTD). Furthermore, CHD8-depleted cells are hypersensitive to drugs that inhibit RNAPII phosphorylation at serine 2, suggesting that CHD8 is required for an early step of the RNAPII transcription cycle.

Hepatitis C in Puerto Rico: a time for public health action

Studies investigating the seroprevalence of HCV infection have been carried out in diverse populations, showing an estimated worldwide prevalence of 3%. A seroprevalence survey conducted among randomly selected non-institutionalized adults aged 21-64 years in San Juan, Puerto Rico in 2001-2002 revealed that 6.3% were positive for HCV antibodies. These data suggest that Puerto Ricans are burdened with a significantly greater prevalence of HCV infection compared to the general United States population aged 20-69 years (0.9%-4.3%). This article illustrates data from different sources that taken together establish the need to start addressing HCV infection in Puerto Rico with prompt and decisive public health actions. Some of these include (1) establish hepatitis C prevention as a priority for state and municipal public health authorities, (2) raise awareness and educate target populations about HCV transmission and prevention, (3) increase clinician awareness of the HCV reporting system and the epidemiology and management of hepatitis C, (4) increase availability of diagnosis and treatment facilities, (5) increase access to effective drug treatment services, and (6) develop appropriate control measures to help reduce continued transmission in correctional settings.

A nucleosome interaction module is required for normal function of Arabidopsis thaliana BRAHMA.

The BRAHMA (BRM) gene encodes the SNF2-type ATPase of the putative Arabidopsis thaliana SWI/SNF chromatin remodelling complex. This family of ATPases is characterized by the presence of a conserved catalytic domain and an arrangement of auxiliary domains, whose functions in the remodelling activity remains unclear. Here, we characterize, at the molecular and functional level, the carboxy-terminal part of Arabidopsis BRM. We have found three DNA-binding regions that bind various free DNA and nucleosomal probes with different specificity. One of these regions contains an AT-hook motif. The carboxy terminus also contains a bromodomain able to bind histones H3 and H4. We propose that this array of domains constitute a nucleosome interaction module that helps BRM to interact with its substrate. We also characterize an Arabidopsis mutant that expresses a BRM protein lacking the last 454 amino acid residues (BRM-DeltaC), encompassing the bromodomain and two of the three DNA-binding activities identified. This mutant displays an intermediate phenotype between those of the wild-type and a null allele mutant, suggesting that the nucleosome interaction module is required for the normal function of BRM but it is not essential for the remodelling activity of BRM-containing SWI/SNF complexes.

Correlates of early sexual activity among Hispanic children in middle adolescence

OBJECTIVE: We analyzed early sexual activity among Hispanic 14 to 15-year-old adolescents residing in a poor neighborhood in Puerto Rico. METHODS: Information from a sample of 325 adolescents was collected from a randomized sample of community households. Logistic regression analysis was used to identify the variables that help explained adolescents' sexual behavior. RESULTS: Adolescents whose parents reported poor communication and poor parent control were more likely to engage in early sexual activity that those peers that did not report this type of family relationship. Adolescents who reported poor parent bonding and lack of discipline were more likely to engage in early sexual relationships. CONCLUSIONS: Intervention and prevention programs need to be aware and address the role of the Hispanic culture in gender differences in early sexual activity in adolescence. If sexual norms related to gender role are changing in Puerto Rico, is a question that needs to be answered in future research.

Risk and protective factors associated with youth violence among secondary school students in a nationally representative sample in Puerto Rico

During the past decade, youth violence has received increasing attention as a major public health issue in Puerto Rico as well as in the United States. This study sought to identify risk and protective factors of youth violence in a representative sample of school adolescents in Puerto Rico. Risk and protective factors were grouped into five domains: individual, family, peer group, school and community. From a total of 2,385 participants, 10.7 reported at least one violent behavior and 3.4 reported two or more violent behaviors. In multiple regression analysis the risk factors identified were male gender, junior grade students, having a favorable attitude towards antisocial behavior, use of ecstasy, involvement with antisocial peers and reporting antisocial parents. Participation in family decisions was the only protective factor associated with violence. Findings from this study could have important implications for the development of preventive programs for the adolescent population in Puerto Rico

Mortality among Hispanic drug users in Puerto Rico

This paper assesses mortality rate for a cohort of drug users in Puerto Rico compared with that of the Island's general population, examining causes of death and estimating relative risk of death. Date and cause of death were obtained from death certificates during 1998. Vital status was confirmed through contact with subjects, family, and friends. HIV/AIDS was the major cause of death (47.7%), followed by homicide (14.6%), and accidental poisoning (6.3%). Females had higher relative risk of death than males in all age categories. Not living with a sex partner and not receiving drug treatment were related to higher mortality due to HIV/AIDS. Drug injection was the only variable explaining relative risk of death due to overdose. Puerto Rico needs to continue developing programs to prevent HIV/AIDS among drug users. Special attention should be given to young women, who appear to be in greatest need of programs to prevent early mortality.

The glucocorticoid receptor interacting protein 1 (GRIP1) localizes in discrete nuclear foci that associate with ND10 bodies and are enriched in components of the 26S proteasome.

The glucocorticoid receptor interacting protein-1 (GRIP1) is a member of the steroid receptor coactivator (SRC) family of transcriptional regulators. Green fluorescent protein (GFP) fusions were made to full-length GRIP1, and a series of GRIP1 mutants lacking the defined regulatory regions and the intracellular distribution of these proteins was studied in HeLa cells. The distribution of GRIP1 was complex, ranging from diffuse nucleoplasmic to discrete intranuclear foci. Formation of these foci was dependent on the C-terminal region of GRIP1, which contains the two characterized transcriptional activation domains, AD1 and AD2. A subpopulation of GRIP1 foci associate with ND10s, small nuclear bodies that contain several proteins including PML, SP100, DAXX, and CREB-binding protein (CBP). Association with the ND10s is dependent on the AD1 of GRIP1, a region of the protein previously described as a CBP-interacting domain. The GRIP1 foci are enriched in components of the 26S proteasome, including the core 20S proteasome, PA28alpha, and ubiquitin. In addition, the irreversible proteasome inhibitor lactacystin induced an increase in the total fluorescence intensity of the GFP-GRIP1 expressing cells, demonstrating that GRIP1 is degraded by the proteasome. These findings suggest the intriguing possibility that degradation of GRIP1 by the 26S proteasome may be a key component of its regulation.

PML and COP1--two proteins with much in common.

The recent discovery that the RING-finger domain is involved in mediating ubiquitin transfer from ubiquitin-conjugating enzymes to substrates have highlighted the importance of protein degradation through the ubiquitin-proteasome pathway in the regulation of different cellular processes. Two RING-finger-containing proteins, the promyelocytic leukemia protein (PML) from mammals and the constitutive photomorphogenic protein (COP1) from plants, show conspicuous similarities in their cellular distribution, dynamics and structure, indicating that they share a related function. Comparison of these two proteins suggests that they are involved in regulating the targeting of nuclear proteins to specific nuclear compartments for degradation through the ubiquitin-proteasome pathway.

Genital diseases in the peruvian dusky dolphin (Lagenorhynchus obscurus).

Ovarian cysts and one ovarian tumour, uterine tumours, vaginal calculi, abscesses of the broad ligament or undetermined testicular lesions were observed in 25 out of 502 female and male dusky dolphins (Lagenorhynchus obscurus) caught off Peru in 1985-1987 or 1993-1994. Tentative or definitive diagnoses included Graafian follicle cysts, luteinized cyst, ovarian parasitic granulomatous inflammation, dysgerminoma, leiomyoma, fibroleiomyoma and chronic fibrino-suppurative inflammation of the broad ligament. All lesions described represented first reports for L. obscurus, and the diagnosis of dysgerminoma was the first in a cetacean. It is also the first time that trematode eggs have been reported in the ovaries of cetaceans and that a vaginal calculus has been encountered in a sexually immature cetacean. The finding of struvite as a major component in two vaginal calculi suggested an infectious aetiology. Of 11 mature females with ovarian tumour or cysts or uterine tumours, only one (9.1%) was pregnant, i.esignificantly less than the expected pregnancy rate (53.3% in a random sample of Peruvian dusky dolphins). Several females with ovarian or uterine lesions and males with aberrant testes were large animals. It is possible that some of these lesions were associated with normal senescence of the reproductive system.

NtcA represses transcription of gifA and gifB, genes that encode inhibitors of glutamine synthetase type I from Synechocystis sp. PCC 6803.

Synechocystis sp. PCC 6803 glutamine synthetase type I (GS) activity is controlled by direct interaction with two inactivating factors (IF7 and IF17). IF7 and IF17 are homologous polypeptides encoded by the gifA and gifB genes respectively. We investigated the transcriptional regulation of these genes. Expression of both genes is maximum in the presence of ammonium, when GS is inactivated. Nitrogen starvation attenuates the ammonium-mediated induction of gifA and gifB as well as the ammonium-mediated inactivation of GS. Putative binding sites for the transcription factor NtcA were identified at -7.5 and -30.5 bp upstream of gifB and gifA transcription start points respectively. Synechocystis NtcA protein binding to both promoters was demonstrated by gel electrophoresis mobility shift assays. Constitutive high expression levels of both genes were found in a Synechocystis NtcA non-segregated mutant (SNC1), which showed a fourfold reduction in the ntcA expression. These experiments indicate a repressive role for NtcA on the transcription of gifA and gifB genes. Our results demonstrate that NtcA plays a central role in GS regulation in cyanobacteria, stimulating transcription of the glnA gene (GS structural gene) and suppressing transcription of the GS inactivating factor genes gifA and gifB.

Glutamine synthetase inactivation by protein-protein interaction.

Glutamine synthetase (GS; EC 6.3.1.2) is the pivotal enzyme of nitrogen metabolism in prokaryotes. Control of bacterial GS activity by reversible adenylylation has provided one of the classical paradigms of signal transduction by cyclic cascades. By contrast, in the present work we show that cyanobacterial GS is controlled by a different mechanism that involves the interaction of two inhibitory polypeptides with the enzyme. Both inactivating factors (IFs), named IF7 and IF17, are required in vivo for complete GS inactivation. Inactive GS-IF7 and GS-IF17 complexes were reconstituted in vitro by using Escherichia coli-expressed purified proteins. Our data suggest that control of GS activity is exerted by regulating the levels of IF7 and IF17.

ras transformation is associated with decreased expression of the brm/SNF2alpha ATPase from the mammalian SWI-SNF complex.

The brm and BRG-1 proteins are mutually exclusive subunits of the mammalian SWI-SNF complex. Within this complex, they provide the ATPase activity necessary for transcriptional regulation by nucleosome disruption. Both proteins were recently found to interact with the p105Rb tumor suppressor gene product, suggesting a role for the mammalian SWI-SNF complex in the control of cell growth. We show here that the expression of brm, but not BRG-1, is negatively regulated by mitogenic stimulation, and that growth arrest of mouse fibroblasts leads to increased accumulation of the brm protein. The expression of this protein is also down-regulated upon transformation by the ras oncogene. Re-introduction of brm into ras transformed cells leads to partial reversion of the transformed phenotype by a mechanism that depends on the ATPase domain of the protein. Our data suggest that increased levels of brm protein favour the withdrawal of the cell from the cycle whereas decreased expression of the brm gene may facilitate cellular transformation by various oncogenes.

Components of the human SWI/SNF complex are enriched in active chromatin and are associated with the nuclear matrix.

Biochemical and genetic evidence suggest that the SWI/SNF complex is involved in the remodeling of chromatin during gene activation. We have used antibodies specific against three human subunits of this complex to study its subnuclear localization, as well as its potential association with active chromatin and the nuclear skeleton. Immunofluorescence studies revealed a punctate nuclear labeling pattern that was excluded from the nucleoli and from regions of condensed chromatin. Dual labeling failed to reveal significant colocalization of BRG1 or hBRM proteins with RNA polymerase II or with nuclear speckles involved in splicing. Chromatin fractionation experiments showed that both soluble and insoluble active chromatin are enriched in the hSWI/SNF proteins as compared with bulk chromatin. hSWI/SNF proteins were also found to be associated with the nuclear matrix or nuclear scaffold, suggesting that a fraction of the hSWI/SNF complex could be involved in the chromatin organization properties associated with matrix attachment regions.

Risk factors for shooting gallery use among drug injectors in Puerto Rico

Needle sharing is one of the principal risk behaviors leading to HIV transmission among injection drug users (IDUs). Shooting galleries, a social context where IDUs rent, share, and borrow needles, are locations usually found near drug markets. This study, which interviewed 1,700 IDUs from May 1989 to June 1990, assesses sociodemographic characteristics and HIV risk behaviors among shooting gallery users in Puerto Rico. Multivariate analyses showed that shooting gallery use is associated with speedball (a concoction of heroin and cocaine) injection, income from illegal activities and previous drug treatment. Shooting gallery users were more likely to rent, share, and borrow needles, and less likely to always use bleach and water to clean needles. Strategies to reduce shooting gallery use among drug injectors are discussed

Mycobacterium tuberculosis infection among crack and injection drug users in San Juan, Puerto Rico

This study aims to determine the prevalence of Mycobacterium tuberculosis infection and its association with HIV and other health risk factors among drug users. A sample of 716 IDUs and crack users were enrolled from community sites. Consenting subjects were tested for HIV serum antibody status. Drug users with an unknown PPD status were administered a PPD skin test and an anergy panel of three antigens (Candida, mumps and tetanus). Overall, 68 (10.3 percent) were reactive to the PPD skin test, 240 (34.7 percent) were HIV positive and 195 (29.5 percent) showed cutaneous anergy. Participants infected with tuberculosis (TB) were more likely to be HIV seropositive, and to have a history of incarceration and residential drug treatment than those not infected. In addition, TB infection was more prevalent among intravenous drug users (IDUs) and shooting gallery managers. These findings suggest that drug users should be considered at high risk for TB and HIV infection. Innovative programs to monitor both infections among drug users are needed to arrest what can become a dual epidemic of HIV and TB in the near future