Minimally invasive blood sampling method for genetic studies on Gopherus tortoises

Método de extracción de sangre mínimamente invasivo para estudios genéticos en tortugas terrestres del género Gopherus La obtención de muestras de tejido de buena calidad es la primera dificultad en cualquier estudio molecular. Esto es especialmente cierto en los estudios de manejo y conservación de la fauna silvestre. En el caso de las tortugas terrestres, la fuente más habitual de ADN son las muestras de sangre obtenidas principalmente de las venas braquial y yugular por contención química, o de individuos conscientes mediante métodos de manipulación y sujeción que pueden causar estrés en el animal. Se requiere una cantidad mínima de sangre para los ensayos del PCR. A continuación, presentamos una técnica mínimamente invasiva que ha resultado eficaz para extraer pequeñas cantidades de sangre apropiadas para realizar análisis genéticos. Además, las muestras obtenidas producen una amplificación de ADN mejor que otras fuentes celulares, como las células epiteliales cloacales. Después de dos años de aplicación en tortugas terrestres silvestres, esta técnica ha demostrado ser inofensiva. Sugerimos que el muestreo de pequeñas cantidades de sangre con esta técnica podría ser útil para otro tipo de análisis, como el seguimiento fisiológico y médico.Obtaining good quality tissue samples is the first hurdle in any molecular study. This is especially true for studies involving management and conservation of wild fauna. In the case of tortoises, the most common sources of DNA are blood samples. However, only a minimal amount of blood is required for PCR assays. Samples are obtained mainly from the brachial and jugular vein after restraining the animal chemically; or from conscious individuals by severe handling methods and clamping. Herein, we present a minimally invasive technique that has proven effective for extracting small quantities of blood, suitable for genetic analyses. Furthermore, the samples obtained yielded better DNA amplification than other cell sources, such as cloacal epithelium cells. After two years of use on wild tortoises, this technique has shown to be harmless. We suggest that sampling a small amount of blood could also be useful for other types of analyses, such as physiologic and medical monitoring.Método de extracción de sangre mínimamente invasivo para estudios genéticos en tortugas terrestres del género Gopherus La obtención de muestras de tejido de buena calidad es la primera dificultad en cualquier estudio molecular. Esto es especialmente cierto en los estudios de manejo y conservación de la fauna silvestre. En el caso de las tortugas terrestres, la fuente más habitual de ADN son las muestras de sangre obtenidas principalmente de las venas braquial y yugular por contención química, o de individuos conscientes mediante métodos de manipulación y sujeción que pueden causar estrés en el animal. Se requiere una cantidad mínima de sangre para los ensayos del PCR. A continuación, presentamos una técnica mínimamente invasiva que ha resultado eficaz para extraer pequeñas cantidades de sangre apropiadas para realizar análisis genéticos. Además, las muestras obtenidas producen una amplificación de ADN mejor que otras fuentes celulares, como las células epiteliales cloacales. Después de dos años de aplicación en tortugas terrestres silvestres, esta técnica ha demostrado ser inofensiva. Sugerimos que el muestreo de pequeñas cantidades de sangre con esta técnica podría ser útil para otro tipo de análisis, como el seguimiento fisiológico y médico

Methodology and Neuromarkers for Cetaceans’ Brains

Cetacean brain sampling may be an arduous task due to the difficulty of collecting and histologically preparing such rare and large specimens. Thus, one of the main challenges of working with cetaceans’ brains is to establish a valid methodology for an optimal manipulation and fixation of the brain tissue, which allows the samples to be viable for neuroanatomical and neuropathological studies. With this in view, we validated a methodology in order to preserve the quality of such large brains (neuroanatomy/neuropathology) and at the same time to obtain fresh brain samples for toxicological, virological, and microbiological analysis (neuropathology). A fixation protocol adapted to brains, of equal or even three times the size of human brains, was studied and tested. Finally, we investigated the usefulness of a panel of 20 antibodies (neuromarkers) associated with the normal structure and function of the brain, pathogens, age-related, and/or functional variations. The sampling protocol and some of the 20 neuromarkers have been thought to explore neurodegenerative diseases in these long-lived animals. To conclude, many of the typical measures used to evaluate neuropathological changes do not tell us if meaningful cellular changes have occurred. Having a wide panel of antibodies and histochemical techniques available allows for delving into the specific behavior of the neuronal population of the brain nuclei and to get a “fingerprint” of their real status

The role of primary pharmacological therapy in acromegaly

BACKGROUND AND OBJECTIVES: Primary pharmacological therapy may be the only viable treatment option for many patients with acromegaly, especially those presenting with advanced disease with large inoperable tumors. Long-acting somatostatin analogs are currently the first-line treatment of choice in this setting, where they provide biochemical control and reduce tumor size in a significant proportion of patients. We herein present a brief overview of the role of primary pharmacological therapy in the treatment of acromegaly within the context of Latin America and support this with a representative case study. CASE DESCRIPTION: A 20 year old male presented with clinical and biochemical evidence of acromegaly. The glucose-suppressed growth hormone (GH) was 5.3 μg/L, his insulin-like growth factor-1(IGF-1) was 3.5 times the ULN and serum prolactin greater than 4,000 μg/L. Pituitary MRI revealed a large and invasive mass, extending superiorly into the optic chiasm and laterally into the left cavernous sinus. He was treated with a combination of octreotide and cabergoline with remarkable clinical improvement, normalization of GH and IGF-1 values and striking shrinkage of the adenoma. CONCLUSION: This case illustrates how effective the pharmacological therapy of acromegaly can be and yet at the same time, raises several important issues such as the need for life-long treatment with costly medications such as the somatostatin analogs. Access to these agents may be limited in regions where resources are restricted and clinicians face challenges in order to make the most efficient use of available options

Speciation in the Emerald Toucanet (*Aulacorhynchus prasinus*) complex

We analyzed genetic variation in the Emerald Toucanet (Aulacorhynchus prasinus), a species complex that ranges primarily along the montane forests of southern and eastern Mexico south to Bolivia. Segments of three mitochondrial DNA genes (cytochrome b, ND2, and ND3) were sequenced for a total of 1,159 base pairs. Using maximum parsimony, maximum likelihood, and Bayesian analysis,we found a set of seven differentiated populations that correspond to clear geographic breaks throughout the highlands of the Neotropics. These genetically distinct populations also correspond with the geographic breaks found in previous analyses of morphological data. Molecular evidence suggests species treatment for four of the Central American clades and three South American clades. Received 19 June 2006, accepted 28 January 2007

Mucosa-associated lymphoid tissue lymphoma of the appendix stump: a case report

Primary lymphomas of the appendix occur in 0.015% of all gastrointestinal lymphomas. The most common manifestation is acute appendicitis secondary to luminal obstruction. The most common is immunophenotype B low-grade non-Hodgkin lymphoma. A 53-year-old male, with a previous three-week surgical history due to acute appendicitis, histopathological report of acute appendicitis and lymphoid hyperplasia. Later, he was admitted, with abdominal pain in the right iliac fossa, an abdominal ultrasound and simple abdominal tomography were performed, with suspicion of residual abscess. Surgical intervention is decided, observing paracecal tumor in the emergency site of the appendix, the tumor is removed. Pathological study that reports an appendicular base infiltrated by mucosa-associated lymphoma. The diagnosis of appendicular tumors is mostly, intraoperatively incidental. It is necessary to have the diagnostic possibility when performing an appendectomy, since it changes the prognosis and treatment of the patient

Pancreatic window and open necrosectomy; a surgical alternative for walled-off pancreatic necrosis in a second level hospital in Mexico

Walled-off pancreatic necrosis is defined as a necrotic collection with a defined wall, which generally occurs in 15% of patients in the fourth week after acute pancreatitis. Actually, open surgery is reserved for selected cases, with minimally invasive treatments such as image-assisted percutaneous drainage or endoscopic ultrasound being the procedures of choice. However, in developing countries the open approach continues to be an effective therapeutic alternative. We present the case of a 47-year-old male patient with no significant history who developed severe acute pancreatitis secondary to hypertriglyceridemia and who later developed walled-off pancreatic necrosis as a late complication. As a treatment, a debridement of the necrotic tissue with marsupialization was performed using the bradley III technique, secondary to the procedure, a pancreatic fistula was developed. After 8 weeks of hospitalization, in which he had a favourable response to surgical treatment, with spontaneous closure of the fistula without complications. Surgical management of late complications of acute pancreatitis remains controversial. Although minimally invasive procedures are the first option nowadays, in developing countries, open necrosectomy remains a good option for the treatment of these types of complications

Apotransferrin-induced recovery after hypoxic/ischaemic injury on myelination

We have previously demonstrated that aTf (apotransferrin) accelerates maturation of OLs (oligodendrocytes) in vitro as well as in vivo. The purpose of this study is to determine whether aTf plays a functional role in a model of H/I (hypoxia/ischaemia) in the neonatal brain. Twenty-four hours after H/I insult, neonatal rats were intracranially injected with aTf and the effects of this treatment were evaluated in the CC (corpus callosum) as well as the SVZ (subventricular zone) at different time points. Similar to previous studies, the H/I event produced severe demyelination in the CC. Demyelination was accompanied by microglial activation, astrogliosis and iron deposition. Ferritin levels increased together with lipid peroxidation and apoptotic cell death. Histological examination after the H/I event in brain tissue of aTf-treated animals (H/I aTF) revealed a great number of mature OLs repopulating the CC compared with saline-treated animals (H/I S). ApoTf treatment induced a gradual increase in MBP (myelin basic protein) and myelin lipid staining in the CC reaching normal levels after 15 days. Furthermore, significant increase in the number of OPCs (oligodendroglial progenitor cells) was found in the SVZ of aTf-treated brains compared with H/I S. Specifically, there was a rise in cells positive for OPC markers, i.e. PDGFRα and SHH+ cells, with a decrease in cleaved-caspase-3+ cells compared with H/I S. Additionally, neurospheres from aTf-treated rats were bigger in size and produced more O4/MBP+ cells. Our findings indicate a role for aTf as a potential inducer of OLs in neonatal rat brain in acute demyelination caused by H/I and a contribution to the differentiation/maturation of OLs and survival/migration of SVZ progenitors after demyelination in vivo