To Poach or Not to Poach an Endangered Species: Elucidating the Economic and Social Drivers Behind Illegal Sea Turtle Hunting in Baja California Sur,Mexico

Despite complete legal protection, improvements in infrastructure, and market conditions that provide easier access to other protein sources, illegal poaching of sea turtles for consumption in Baja California Sur (BCS), Mexico remains a major threat to their recovery. Few studies have focused on understanding the economic and social drivers behind this activity, which is fundamental to determining best practices for discouraging it. From June 2007 to April 2008 we conducted eight in-depth, semi-structured interviews with sea turtle poachers at five coastal communities in BCS to determine the drivers influencing them. The most prevalent reasons for illegal poaching were direct economic benefits, lack of law enforcement and ease of escape from or bribery of authorities, and strong family tradition. Our results suggest that to reduce illegal poaching it will be necessary to better enforce existing environmental laws, reduce social acceptance of sea turtle hunting throughout the region, educate fishers on the ecological importance of sea turtles, and show fishers direct economic benefits from non-consumptive use of sea turtles, such as ecotourism.     

Diagnosis of mitochondrial disorders applying massive pyrosequencing

Mitochondrial disorders are a frequent cause of neurological disability affecting children and adults. Traditionally, molecular diagnosis of mitochondrial diseases was mostly accomplished by the use of Sanger sequencing and PCR–RFLP. However, there are particular drawbacks associated with the use of these methods. Recent multidisciplinary advances have led to new sequencing methods that may overcome these limitations. Our goal was to explore the use of a next generation sequencing platform in the molecular diagnosis of mitochondrial diseases reporting our findings in adult patients that present with a clinical-pathological diagnosis of a mitochondrial encephalomyopathy. Complete genomic sequences of mitochondrial DNA were obtained by 454 massive pyrosequencing from blood samples. The analysis of these sequences allowed us to identify two diagnostic pathogenic mutations and 74 homoplasmic polymorphisms, useful for obtaining high-resolution mitochondrial haplogroups. In summary, molecular diagnosis of mitochondrial disorders could be efficiently done from readily accessible samples, such as blood, with the use of a new sequencing platform.     

Early evidence of lymphoproliferative disorder: post-transplant monitoring of Epstein-Barr infection in adult and pediatric patients

Transplant patients are at high risk of post-transplant lymphoproliferative disorder (PTLD). A strong correlation between Epstein-Barr virus (EBV) and PTLD is observed in pediatric patients with primary infection after transplant. Because many patients have responded to reversal of immunosuppressive therapy, an early identification of EBV is essential for the reduction of immunosuppression and/or introduction of antiviral therapy to prevent PTLD. Polymerase chain reaction (PCR) is a specific and sensitive method to identify EBV DNA in blood. The aim of our study was to establish a protocol for monitoring EBV infection in transplanted patients for early identification those at high risk of PTLD. Viral presence in peripheral blood leukocytes (PBL) and serum samples was revealed by Nested PCR; positive specimens were quantified with Real Time PCR (RT-PCR). DNA in PBL was observed in 12 cases and 6 showed EBV in sera. Quantitative analysis showed a wide range of EBV DNA copies in leukocytes that were higher than in sera. Two patients displayed high viral load values in both PBL and sera associated with clinical evidence of PTLD. Our data suggest that the study of the EBV load represents an essential approach in the diagnosis of PTLD and the analysis of serum samples could provide useful information in the post-transplant monitoring of high-risk patients.     

The crystal structure of UMP kinase from Bacillus anthracis (BA1797) reveals an allosteric nucleotide-binding site

Uridine monophosphate (UMP) kinase is a conserved enzyme that catalyzes the ATP-driven conversion of uridylate monophosphate into uridylate diphosphate, an essential metabolic step. In prokaryotes, the enzyme exists as a homohexamer that is regulated by various metabolites. Whereas the enzymatic mechanism of UMP kinase (UK) is well-characterized, the molecular basis of its regulation remains poorly understood. Here we report the crystal structure of UK from Bacillus anthracis (BA1797) in complex with ATP at 2.82 Å resolution. It reveals that the cofactor, in addition to binding in the active sites, also interacts with separate binding pockets located near the center of the hexameric structure. The existence of such an allosteric binding site had been predicted by biochemical studies, but it was not identified in previous crystal structures of prokaryotic UKs. We show that this putative allosteric pocket is conserved across different bacterial species, suggesting that it is a feature common to bacterial UKs, and we present a structural model for the allosteric regulation of this enzyme.     

Atomic snapshots of an RNA packaging motor reveal conformational changes linking ATP hydrolysis to RNA translocation

Many viruses package their genome into preformed capsids using packaging motors powered by the hydrolysis of ATP. The hexameric ATPase P4 of dsRNA bacteriophage phi12, located at the vertices of the icosahedral capsid, is such a packaging motor. We have captured crystallographic structures of P4 for all the key points along the catalytic pathway, including apo, substrate analog bound, and product bound. Substrate and product binding have been observed as both binary complexes and ternary complexes with divalent cations. These structures reveal large movements of the putative RNA binding loop, which are coupled with nucleotide binding and hydrolysis, indicating how ATP hydrolysis drives RNA translocation through cooperative conformational changes. Two distinct conformations of bound nucleotide triphosphate suggest how hydrolysis is activated by RNA binding. This provides a model for chemomechanical coupling for a prototype of the large family of hexameric helicases and oligonucleotide translocating enzymes.     

alpha-Glycerylphosphorylethanolamine rescues astrocytes from mitochondrial impairment and oxidative stress induced by amyloid beta-peptides

The present work shows that alpha-glycerylphosphorylethanolamine (alpha-GPE) is effective in recovering astrocytes from mitochondrial membrane integrity and potential derangement and cellular oxidative stress that occur under amyloid beta-peptides-induced reactive gliosis.alpha-Glycerylphosphorylethanolamine (alpha-GPE), a new compound with nootropic properties, known to improve in vivo the learning and memory processes, has been tested for its protective properties on an in vitro model of degeneration. Rat primary astrocytic cultures treated with two amyloid-derived peptides, Abeta((1-40)) and Abeta(3(pE)-42), showed a marked reduction of the mitochondrial redox activity and membrane potential, together with an increase of oxidative species production. Plasma membrane lipid peroxidation (LPO) as well as generation of peroxides is greatly increased under Abeta-peptides toxicity. These features, typical of the reactive gliosis that accompanies neuronal degeneration, were readily recovered by pretreatment with alpha-GPE. alpha-GPE, likely improving the fluidity of cell membrane, has the potential to recover astrocytes from the general redox derangement induced by different amyloid fragments and possibly to protect from inflammation, gliosis and neurodegeneration. This is the first evidence of an antioxidant effect of the ethanolamine derivative on a rat model of chronic gliosis.     

Phage display for the production of human monoclonal antibodies against human pathogens

In the last decade an increasing number of antibodies have made their way from the research benchtops into the clinics and many more are currently under clinical trial. Among monoclonal antibody-producing techniques, phage-display is undoubtedly the most effective and versatile. Cloning of the entire humoral repertoire derived from an infected patients into a phage display vector allows not only the simple generation of monoclonal antibodies of desired specificity, but also the molecular dissection of the antibody response itself. Generation of large panels of human monoclonal antibodies against human pathogens could open new perspectives in understanding the interplay between the infectious agent and the infected host providing tools for the prevention and the therapy of human communicable diseases. In this paper the basic principles of the phage-display approach as well as its most recent applications are reviewed.     

A retrospective hormonal and immunohistochemical evaluation of 47 acromegalic patients: prognostic value of preoperative plasma prolactin.

This study was performed to investigate the correlations between preoperative prolactin (PRL) plasma values, immunohistochemical picture and the clinical course in growth hormone (GH) secreting pituitary adenomas. In 47 patients (19 males and 28 females; mean age 40 years; range 13 - 70 years), we measured GH, IGF-1 and prolactin plasma values both before and after transsphenoidal surgery, and basal IGF-1 and GH after an oral glucose tolerance test (OGTT) during four years of follow-up. We considered those patients as "controlled" who presented an undetectable growth hormone after OGTT (GH < 1 microg/l), IGF-I plasma values in the normal range, matched for age and sex, and no clinical activity or neuroradiological recurrence after a four-year follow-up. We considered patients as "poorly controlled" who still showed elevated GH and IGF-I plasma levels, uninhibited GH after OGTT (GH > 1 microg/l), presence of clinical activity and/or radiological signs of adenoma recurrence, even if a reduction of tumor size had been demonstrated. RESULTS: Controlled patients (n = 22) exhibited mean preoperative PRL levels (+/- SEM) lower than the group of poorly controlled (n = 25) ones (21.40 +/- 5.51 vs. 38.44 +/- 5.16 microg/l; p < 0.03). From 3 to 12 months after surgery, postoperative PRL levels were also lower in the controlled patients compared to the poorly controlled ones (8.31 +/- 1.20 vs. 25.32 +/- 3.20 microg/l; p < 0.0001). Eighty percent (20/25) of poorly controlled patients showed both PRL and GH positivity after immunostaining. Only 3/22 (13.6 %) of controlled patients showed the same double positivity. In conclusion, preoperative hyperprolactinemia identifies a group of acromegalic patients at elevated risk of disease persistence after surgery. We hypothesize that most of these high-risk patients may have more aggressive mixed GH-PRL secreting adenomas.