Recombinant adenovirus delivery of calreticulin-ESAT-6 produces an antigen-specific immune response but no protection against a Mycobacterium tuberculosis challenge.

Bacillus Calmette-Guerin (BCG) has failed to efficaciously control the worldwide spread of the disease. New vaccine development targets virulence antigens of Mycobacterium tuberculosis that are deleted in Mycobacterium bovis BCG. Immunization with ESAT-6 and CFP10 provides protection against M. tuberculosis in a murine infection model. Further, previous studies have shown that calreticulin increases the cell-mediated immune responses to antigens. Therefore, to test whether calreticulin enhances the immune response against M. tuberculosis antigens, we fused ESAT-6 to calreticulin and constructed a recombinant replication-deficient adenovirus to express the resulting fusion protein (AdCRT-ESAT-6). The adjuvant effect of calreticulin was assayed by measuring cytokine responses specific to ESAT-6. Recombinant adenovirus expressing the fusion protein produced higher levels of interferon-γ and tumour necrosis factor-α in response to ESAT-6. This immune response was not improved by the addition of CFP-10 to the CRT-ESAT-6 fusion protein (AdCRT-ESAT-6-CFP10). Mice immunized with these recombinant adenoviruses did not decrease the mycobacterial burden after low-dose aerosol infection with M. tuberculosis. We conclude that calreticulin can be used as an adjuvant to enhance the immune response against mycobacterial antigens, but it is not enough to protect against tuberculosis.

[Immunofluorescence mapping for diagnosis of congenital epidermolysis bullosa]. / Mapeo por inmunofluorescencia para el diagnóstico de epidermólisis ampollosa congénita.

The tools for diagnosis of epidermolysis bullosa have advanced greatly since Hintner's group introduced antigen mapping as a diagnostic test for this family of genodermatoses. Monoclonal or polyclonal antibodies raised against some of the specific proteins found in the epidermis and basement membrane of the epidermis have allowed 4 types of epidermolysis bullosa de be identified and all variants to be classified. When a newborn baby presents with blisters, many conditions are implicated in the differential diagnosis. Examination under an optical microscope can suggest epidermolysis bullosa, but immunofluorescence mapping and electron microscopy are required for confirmation of the diagnosis and further classification of congenital epidermolysis bullosa. This article explains the importance of immunofluorescence antigen mapping and describes the methods employed for classification and subclassification of epidermolysis bullosa.

Karwinskia humboldtiana (buckthorn) fruit causes central nervous system damage during chronic intoxication in the rat.

Karwinskia humboldtiana fruit (Kh) causes a neurological disorder 3-4 weeks after ingestion, characterized by flaccid, symmetrical, ascending paralysis, similar to the Guillain-Barre syndrome. In this polyneuropathy the lesion (demyelization) in peripheral nerves has been described in several animal species, both in acute and in chronic intoxication. However, no reports exist about the presence of lesions in the Central Nervous System (CNS), in chronic intoxication. We considered it important to evaluate, with histological techniques, the possible presence of lesions in the brain, by using a model of chronic intoxication that reproduces the same stages present in the human intoxication, to better understanding of this pathological process. In our present work we fed the ground Kh fruit to Wistar rats and samples of brain, cerebellum, and pons were embedded in paraffin. Sections were stained with Hematoxylin & Eosin (HE) and special stains for nerve tissue. Histopathological changes were evaluated in the CNS through the different stages of the polyneuropathy and comparison to a control group. With this methodology, we found lesions in the motor pathway. This is the first report about the presence of neuronal damage caused by Kh in the Central Nervous System in chronic intoxication.

Folate levels and N(5),N(10)-methylenetetrahydrofolate reductase genotype (MTHFR) in mothers of offspring with neural tube defects: a case-control study.

BACKGROUND: Neural tube defects (NTDs) have been associated with biochemical factors involved in the conversion of homocysteine to methionine as folate deficiency and the mutation 677T in the N(5),N(10)-methylenetetrahydrofolate reductase gene (MTHFR). METHODS: A case-control study was performed to detect this mutation in 38 unrelated women with NTD deceased products and 31 mothers without antecedents of NTD offspring. All products were born in Nuevo León (northeastern Mexico) during 1997. Erythrocyte and plasmatic folate levels and the genotype of the 677 polymorphism at the MTHFR locus were analyzed in both groups. RESULTS: Although no significant differences were found in mean blood folate levels, the percentage of women in the case group with erythrocyte folate levels <160 ng/mL was significantly higher than in the control group (75 vs. 51.2%, p <0.05). The proportion of women with plasma folate levels <3.5 ng/mL was higher in the case group (16.2 vs. 0%, p <0.01). Genotype analysis demonstrated a significantly higher proportion of 677T homozygous mothers with NTD products (39.6 vs. 9.1%, p <0.05). Allele frequencies for the 677T mutation were 0.55 and 0.36 for cases and controls, respectively. The odds ratio (OR) for having a NTD product was 6.1 (95%, CI 1.56-23.6) for homozygous 677T mothers vs. homozygous 677C and heterozygous mothers. Significantly low levels of erythrocyte folate were found in the 677C homozygous case group and in plasma folate in the 677C/677T heterozygous case mothers. CONCLUSIONS: Our study suggests that folate deficiency and MTHFR unfavorable genotype in mothers are important risk factors for severe NTD phenotype in our population.

[Diagnosis of thrombosis of cavernous sinus in magnetic resonance imaging]. / Diagnóstico de trombosis del seno cavernoso en imagen por resonancia magnética.

The diagnosis of the cavernous sinus thrombosis requires a careful clinical evaluation and appropriate radiological methods. In this report we describe the magnetic resonance imaging findings in a patient with the clinical diagnosis of cavernous sinus disorder and its correlation with orbital phlebography and histopathologic studies.

Secondary malignant transformation of testicular teratomas: case series and literature review.

BACKGROUND: Teratomas are a spectrum of neoplasms that can undergo malignant transformation. In the World Health Organization (WHO) classification of tumors, this entity was classified as «teratoma with somatic-type malignancy¼, was defined as a malignant neoplasm of non-germinal phenotype that originates in a teratoma. MATERIALS AND METHODS: We present a serie of nine cases of testicular teratomas with secondary malignant transformation. From January 1995 to December 2011, we found a total of 306 cases of testicular tumors. Mixed germ cell tumors were the most frequently diagnosed malignancy with 45.7%. RESULTS: Teratoma with secondary malignant transformation, represented 2.9% of all germinal tumors. Five cases originated within a mixed germ cell tumor, two cases from mature teratomas, and two from immature teratomas. The predominante malignant somatic component were sarcomas; two cases of chondrosarcoma, one rhabdomyosarcoma, and one case showing foci of chondrosarcoma and rhabdomyosarcoma. The case of osteosarcoma is notable for its rarity. Two cases showed epithelial malignancy in the form of an adenocarcinoma, and finally, two cases were primitive neuroectodermal tumors. At the time of diagnosis, five patients had metastases. CONCLUSION: The transformation of germ cell tumors to somatic type malignancies is rare. The malignant component can originate from any of the three germ lines. These tumors are resistant to standard chemotherapy for a germ cell tumor and the clinical stage is the most important prognostic factor. At our institution, the malignant component that appeared most frequently was chondrosarcoma.

Disseminated paracoccidioidomycosis in a Southern two-toed sloth (Choloepus didactylus).

A Southern two-toed sloth (Choloepus didactylus), originally acquired from French Guiana, died while maintained in quarantine in a pet store in Monterrey, Mexico. Large yeast cells with multiple buds compatible with Paracoccidioides brasiliensis were observed in disseminated granulomatous lesions in the lungs, liver, spleen and kidney. Transmission electron microscopical examination supported the diagnosis. This is the first report of paracoccidioidomycosis in a two-toed sloth.

Ultrastructure of the seminiferous epithelium of ethyl methanesulphonate-treated mouse.

Ethyl methanesulphonate (EMS) is a mutagenic alkylating agent that induces marked elevations of sperm abnormalities in mice. In this paper, we report the ultrastructural findings on the morphology of the seminiferous epithelium of mice resulting from EMS administration. Eight- to twelve-weeks-old male mice were injected intraperitoneally with EMS at 200 mg kg(-1) body weight daily for five consecutive days. Analysis of smears of epididymis and semi-thin sections of testes revealed that the more suitable specimens for the ultrastructural analysis were tissues of mice killed at the third week, following EMS administration. At this time, the spermatid was the damaged cell type. Abnormalities were mainly observed in the morphology of the nucleus, the acrosome, chromatin distribution and in the arrangement of the cytoplasmic microtubules, and binucleated spermatids were also observed. EMS has the capacity to penetrate the blood-testis barrier, and thus it can damage post-meiotic spermatogenic cells. However, morphological abnormalities could be the consequence of damage exerted on the differentiated spermatogonia stage, the most sensitive spermatogenic cell to the action of chemical agents or drugs. Our findings contribute to elucidate the action mechanism of the damage exerted by EMS administration on the germinal male cells.

Transformación maligna secundaria de teratomas testiculares: serie de casos y revisión de la literatura / Secondary malignant transformation of testicular teratomas: Case series and literature review

Antecedentes: Los teratomas son un espectro de neoplasias que pueden sufrir una transformación maligna. En la clasificación de la Organización Mundial de la Salud (OMS) de los tumores esta entidad fue clasificada como «teratoma con malignidad de tipo somático», se definió como una neoplasia maligna de fenotipo no germinal que se origina en un teratoma. Materiales y métodos: Se presenta una serie de 9 casos de teratomas testiculares con transformación maligna secundaria. Entre enero de 1995 y diciembre de 2011 encontramos un total de 306 casos de tumores testiculares. Los tumores de células germinales mixtas fueron el tumor maligno más frecuentemente diagnosticado con un 45,7%. Resultados: El teratoma con transformación maligna secundaria representó el 2,9% de todos los tumores germinales. Cinco casos se originaron dentro de un tumor de células germinales mixtas, 2 casos de teratomas maduros y 2 de teratomas inmaduros. El componente somático maligno predominante eran los sarcomas; 2 casos de condrosarcoma, uno de rabdomiosarcoma y un caso que muestra focos de condrosarcoma y rabdomiosarcoma. El caso de osteosarcoma se destaca por su rareza. Dos casos mostraron malignidad epitelial en la forma de un adenocarcinoma y, finalmente, 2 casos eran tumores neuroectodérmicos primitivos. En el momento del diagnóstico 5 pacientes tenían metástasis. Conclusión: La transformación de los tumores de células germinales en tumores malignos de tipo somático es poco común. El componente maligno puede proceder de cualquiera de las 3 líneas germinales. Estos tumores son resistentes a la quimioterapia estándar para un tumor de células germinales, y el estadio clínico es el factor pronóstico más importante. En nuestra institución el componente maligno que apareció con mayor frecuencia fue el condrosarcoma

Background: Teratomas are a spectrum of neoplasms that can undergo malignant transformation. In the World Health Organization (WHO) classification of tumors, this entity was classified as «teratoma with somatic-type malignancy», was defined as a malignant neoplasm of non-germinal phenotype that originates in a teratoma. Materials and methods: We present a serie of nine cases of testicular teratomas with secondary malignant transformation. From January 1995 to December 2011, we found a total of 306 cases of testicular tumors. Mixed germ cell tumors were the most frequently diagnosed malignancy with 45.7%. Results: Teratoma with secondary malignant transformation, represented 2.9% of all germinal tumors. Five cases originated within a mixed germ cell tumor, two cases from mature teratomas, and two from immature teratomas. The predominante malignant somatic component were sarcomas; two cases of chondrosarcoma, one rhabdomyosarcoma, and one case showing foci of chondrosarcoma and rhabdomyosarcoma. The case of osteosarcoma is notable for its rarity. Two cases showed epithelial malignancy in the form of an adenocarcinoma, and finally, two cases were primitive neuroectodermal tumors. At the time of diagnosis, five patients had metastases. Conclusion: The transformation of germ cell tumors to somatic type malignancies is rare. The malignant component can originate from any of the three germ lines. These tumors are resistant to standard chemotherapy for a germ cell tumor and the clinical stage is the most important prognostic factor. At our institution, the malignant component that appeared most frequently was chondrosarcoma

Mapeo por inmunofluorescencia para el diagnóstico de epidermólisis ampollosa congénita / Immunofluorescence Mapping for Diagnosis of Congenital Epidermolysis Bullosa

Las herramientas para el diagnóstico en las epidermólisis ampollosas (EA) han tenido un gran avance desde que Hintner et al, introdujeron el mapeo antigénico como prueba diagnóstica en este grupo de genodermatosis. La utilización de anticuerpos monoclonales/policlonales dirigidos contra algunas de las proteínas específicas que conforman la epidermis y la membrana basal epidérmica, han servido para clasificar los 4 tipos de epidermólisis ampollosa y subclasificar todas sus variantes. Ante la presencia de un recién nacido con ampollas surgen diagnósticos diferenciales múltiples, en donde la microscopia de luz orienta el diagnostico de epidermólisis ampollosa. Sin embargo, el mapeo por inmunofluorescencia y la microscopia electrónica permiten confirmar y clasificar a las epidermólisis ampollosas congénitas. En este artículo, se explica la importancia y metodología para desarrollar la técnica de mapeo antigénico por inmunofluorescencia, con el propósito de clasificar y subclasificar las epidermólisis ampollosas (AU)

The tools for diagnosis of epidermolysis bullosa have advanced greatly since Hintner's group introduced antigen mapping as a diagnostic test for this family of genodermatoses. Monoclonal or polyclonal antibodies raised against some of the specific proteins found in the epidermis and basement membrane of the epidermis have allowed 4 types of epidermolysis bullosa de be identified and all variants to be classified. When a newborn baby presents with blisters, many conditions are implicated in the differential diagnosis. Examination under an optical microscope can suggest epidermolysis bullosa, but immunofluorescence mapping and electron microscopy are required for confirmation of the diagnosis and further classification of congenital epidermolysis bullosa. This article explains the importance of immunofluorescence antigen mapping and describes the methods employed for classification and subclassification of epidermolysis bullosa (AU)