Organ-Specific Expression of IL-1 Receptor Results in Severe Liver Injury in Type I Interferon Receptor Deficient Mice.

Upon treatment with polyinosinic:polycytidylic acid [poly(I:C)], an artificial double-stranded RNA, type I interferon receptor-deficient (IFNAR-/-) mice develop severe liver injury seen by enhanced alanine aminotransferase (ALT) activity in the serum that is not observed in their wildtype (WT) counterparts. Recently, we showed that liver injury is mediated by an imbalanced expression of interleukin (IL)-1ß and its receptor antagonist (IL1-RA) in the absence of type I IFN. Here we show that despite comparable expression levels of IL-1ß in livers and spleens, spleens of poly(I:C)-treated IFNAR-/- mice show no signs of injury. In vitro analyses of hepatocytes and splenocytes revealed that poly(I:C) had no direct toxic effect on hepatocytes. Furthermore, expression levels of cytokines involved in other models for liver damage or protection such as interferon (IFN)-γ, transforming growth factor (TGF)-ß, IL-6, IL-10, IL-17, and IL-22 were comparable for both organs in WT and IFNAR-/- mice upon treatment. Moreover, flow cytometric analyses showed that the composition of different immune cells in livers and spleens were not altered upon injection of poly(I:C). Finally, we demonstrated that the receptor binding IL-1ß, IL1R1, is specifically expressed in livers but not spleens of WT and IFNAR-/- mice. Accordingly, mice double-deficient for IFNAR and IL1R1 developed no liver injury upon poly(I:C) treatment and showed ALT activities comparable to those of WT mice. Collectively, liver injury is mediated by the organ-specific expression of IL1R1 in the liver.

Twin Reversed Arterial Perfusion Sequence (TRAPS): An Illustrative Series of 13 Cases.

Twin reversed-arterial-perfusion sequence (TRAPS) is a rare and severe complication of monochorionic twin pregnancies. It usually occurs in the setting of monochorionic placentation, when the heart of a normal appearing twin serves as the pump for one or more dysmorphic twins whose head, thoracic organs, and upper extremities do not fully develop or do not develop at all and thus lack cardiac activity. Anomalous vascular placental architecture causes a shift in arterial flow towards the acardiac twin(s). The exact physiopathologic mechanisms that lead to this devastating phenomenon are not well known. We reviewed the maternal history and the surgical pathology reports of the fetuses and placentas of 13 different cases of TRAPS that were collected in a 23-year study period at a single institution. Herein we summarize the characteristic findings and illustrate specific mechanical feto-placental circulation issues that appear to be instrumental in the development of TRAPS.

Representative Sinusoids for Hepatic Four-Scale Pharmacokinetics Simulations.

The mammalian liver plays a key role for metabolism and detoxification of xenobiotics in the body. The corresponding biochemical processes are typically subject to spatial variations at different length scales. Zonal enzyme expression along sinusoids leads to zonated metabolization already in the healthy state. Pathological states of the liver may involve liver cells affected in a zonated manner or heterogeneously across the whole organ. This spatial heterogeneity, however, cannot be described by most computational models which usually consider the liver as a homogeneous, well-stirred organ. The goal of this article is to present a methodology to extend whole-body pharmacokinetics models by a detailed liver model, combining different modeling approaches from the literature. This approach results in an integrated four-scale model, from single cells via sinusoids and the organ to the whole organism, capable of mechanistically representing metabolization inhomogeneity in livers at different spatial scales. Moreover, the model shows circulatory mixing effects due to a delayed recirculation through the surrounding organism. To show that this approach is generally applicable for different physiological processes, we show three applications as proofs of concept, covering a range of species, compounds, and diseased states: clearance of midazolam in steatotic human livers, clearance of caffeine in mouse livers regenerating from necrosis, and a parameter study on the impact of different cell entities on insulin uptake in mouse livers. The examples illustrate how variations only discernible at the local scale influence substance distribution in the plasma at the whole-body level. In particular, our results show that simultaneously considering variations at all relevant spatial scales may be necessary to understand their impact on observations at the organism scale.

Possible Genetic Origin of Limb-Body Wall Complex.

Limb body wall complex (LBWC) is characterized by multiple severe congenital malformations including an abdominal and/or thoracic wall defect covered by amnion, a short or absent umbilical cord with the placenta almost attached to the anterior fetal wall, intestinal malrotation, scoliosis, and lower extremity anomalies. There is no consensus about the etiology of LBWC and many cases with abnormal facial cleft do not meet the requirements for the true complex. We describe a series of four patients with LBWC and other malformations in an attempt to explain their etiology. There are several reports of fetuses with LBWC and absent gallbladder and one of our patients also had polysplenia. Absent gallbladder and polysplenia are associated with laterality genes including HOX, bFGF, transforming growth factor beta/activins/BMP4, WNT 1-8, and SHH. We postulate that this severe malformation may be due to abnormal genes involved in laterality and caudal development.

Bilateral renal dysplasia, nephroblastomatosis, and bronchial stenosis. A new syndrome?

Bilateral nephroblastomatosis (NB) is an uncommon renal anomaly characterized by multiple confluent nephrogenic rests scattered through both kidneys, with only a limited number of cases reported in the medical literature. Some of these children may have associated either Perlman or Beckwith-Wiedemann syndrome and others do not demonstrate syndromic features. We report a full-term boy with anteverted nose, bilateral bronchial stenosis due to lack of cartilage, bilateral obstructive renal dysplasia and NB with glomeruloid features. The infant had visceromegaly, but neither gigantism nor hemihypertrophy. Immunohistochemistry for PAX2 (Paired box gene-2) and WT-1 (Wilms Tumor 1) were strongly positive in the areas of NB. GLEPP-1 (Glomerular Epithelial Protein) did not stain the areas of NB with a glomeruloid appearance, but was positive in the renal glomeruli as expected. We found neither associated bronchial stenosis nor the histology of NB resembling giant glomeruli in any of the reported cases of NB.

Quantification of sickle cells in the peripheral smear as a marker of disease severity.

Blinded readers examined peripheral smears of 108 children with steady sickle cell (SC) disease and controls by counting ten 100 × microscope fields and calculating percent of irreversible and reversible SC from total red cell population SC index (SCI). SCI was correlated to disease severity, and transfusion, hydroxyurea, or neither. Controls had a mean of 0.28% SC (range 0-0.64). Children with hemoglobin SS had a mean SCI of 5.12% ± 5.37 (range 0-30). SCI increased 0.33% with each increasing year (p < 0.0001). Patients with SCI >0.64 were 3.32 times as likely to experience clinical complications (p = 0.0124). Although blood transfusions and hydroxyurea decreased percent of SC, 72% treated patients had SCI >0.64, correlating with persistent sickling. This standardized method quantifies SC in peripheral smears. Percent of SC increased with age and correlated with disease severity, especially hemolytic complications, providing readily available information with minimal or no extra cost.

Bridging the gaps between the histopathological and demographic risk factors of preterm birth in a unique Miami inner-city population.

We aim to identify the link between placental histological findings and obstetric reports to determine possible risk factors of spontaneous preterm birth (SPTB). We prospectively ascertained birth records and outcomes from all deliveries in our hospital in 1 year. Records were used to determine and stratify for either full-term or preterm [spontaneous or indicated (I)] deliveries. We analyzed for risk factor association using χ(2) tests and common odds ratio estimates (SPSS v21.0). Our cohort totaled 6088 deliveries: 236 IPTB, 43 SPTB, and 5809 term births. Largely Hispanic, we determined race, parity, prenatal care access, preeclampsia, gestational diabetes, and BMI to be highly associated with SPTB (p < 0.01). Histologically, placentas of women with SPTB were twice as likely to have chronic villitis. We found that chronic villitis is associated with SPTB. Results of this study can be used in increasing the understanding of SPTB.

Autoimmune hepatitis and primary sclerosing cholangitis in children and adolescents.

UNLABELLED: Clinical presentation and histopathology of autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC) overlap syndrome (OS) are similar, but their management is different. We conducted a pediatric retrospective cross-sectional study of 34 patients with AIH and PSC. AIH had female predominance (74%) and was lower in PSC (45%). There was a trend toward higher frequency of blacks in PSC/OS (55%) compared to Caucasians (36%) and Hispanics (9%), but not race differences in AIH. Inflammatory bowel disease (IBD) was present in 75% of PSC/OS. Plasma cells were not specific for AIH (found in 42% of PSC). Concentric fibrosis was not reliable for PSC as was found in 46% of AIH. CONCLUSION: A combination of clinical history, laboratory tests, imaging studies and liver biopsy are required to confirm and properly treat AIH and PSC. Liver biopsy should be used to grade severity and disease progression, but cannot be used alone to diagnose these conditions.

Evaluación de los medios de enseñanza utilizados para el aprendizaje de la morfofisiología del sistema osteomioarticular / Assessment of the use of teaching aids to teach the morphology and physiology of Osteomyoarticular System

Introducción: los medios de enseñanza utilizados en el proceso de enseñanza-aprendizaje de la morfofisiología deben ser perfeccionados para consolidar los conocimientos de las ciencias básicas en la carrera de Medicina. Objetivo: evaluar la utilización de los medios, en el proceso enseñanza-aprendizaje de la morfofisiología del sistema osteomioarticular en el primer año de la carrera de Medicina, de la Facultad de Ciencias Médicas Dr."Ernesto Che Guevara de la Serna", Pinar del Río. Material y métodos: se utilizaron los métodos empíricos, teóricos y procedimientos de la estadística descriptiva. Fueron considerados como universo los profesores que imparten o han impartido la propia asignatura (n=6), así como los profesores asesores de la disciplina(n=4) y estudiantes de primer año de la carrera de Medicina del curso 2010-2011, que recibieron la asignatura Morfofisiología Humana II (n=161). Resultados: se encontró que no existe una correspondencia entre la organización de los contenidos ubicados en el CD y los que están incluidos actualmente en el programa de la asignatura, criterios de expertos, profesores y estudiantes, con relación a las dificultades que existen con los medios, lo que se comprobó en la generalidad de las respuestas. Conclusiones: los medios de enseñanza utilizados para el aprendizaje de la morfofisiología del sistema osteomioarticular no responden a la necesidad de un aprendizaje interactivo, donde se construya el conocimiento, y el estudiante pueda desarrollar sus propias estrategias de aprendizaje, por lo que se hace necesario perfeccionar los mismos.

Introduction: the use of teaching aids on the teaching-learning process of morphology and physiology subjects should be improved to strengthen knowledge of basic sciences in Medical Studies. Objective: to assess the use of teaching aids on the teaching-learning process of morphology and physiology of the osteomyoarticular system on 1st academic year of medical studies at "Dr. Ernesto Guevara de la Serna" Medical School. Pinar del Rio. Material and Methods: empirical and theoretical methods were used along with descriptive statistics procedures. Professors of the subjects were taken as a target group (n=6), as well as the consultants of the subject (n=4) and students of the 1st academic year of medical studies receiving the subject of Human Morphology and Physiology II (n=161) during 2010-2011. Results: no correspondence was found regarding the organization of contents on CDs and the ones currently included in the syllabus of the subject, criteria of experts, professors and students in relation to the difficulties with the teaching aids, which was verified in the majority of the responses. Conclusions: the teaching aids used to teach the morphology and physiology of osteomyoarticular system give no responses to an interactive learning, where knowledge must be built and for the students to develop their own learning strategies, that is why it is necessary to better the teaching aids.

Evaluación de los medios de enseñanza utilizados para el aprendizaje de la morfofisiología del sistema osteomioarticular / Assessment of the use of teaching aids to teach the morphology and physiology of Osteomyoarticular System

Los medios de enseñanza utilizados en el proceso de enseñanza-aprendizaje de la morfofisiología deben ser perfeccionados para consolidar los conocimientos de las ciencias básicas en la carrera de Medicina. El objetivo fue evaluar la utilización de los medios, en el proceso enseñanza-aprendizaje de la morfofisiología del sistema osteomioarticular en el primer año de la carrera de Medicina, de la Facultad de Ciencias Médicas Dr. Ernesto Che Guevara de la Serna, Pinar del Río. Se utilizaron los métodos empíricos, teóricos y procedimientos de la estadística descriptiva. Fueron considerados como universo los profesores que imparten o han impartido la propia asignatura (n=6), así como los profesores asesores de la disciplina(n=4) y estudiantes de primer año de la carrera de Medicina del curso 2010-2011, que recibieron la asignatura Morfofisiología Humana II (n=161). Se encontró que no existe una correspondencia entre la organización de los contenidos ubicados en el CD y los que están incluidos actualmente en el programa de la asignatura, criterios de expertos, profesores y estudiantes, con relación a las dificultades que existen con los medios, lo que se comprobó en la generalidad de las respuestas. Los medios de enseñanza utilizados para el aprendizaje de la morfofisiología del sistema osteomioarticular no responden a la necesidad de un aprendizaje interactivo, donde se construya el conocimiento, y el estudiante pueda desarrollar sus propias estrategias de aprendizaje, por lo que se hace necesario perfeccionar los mismos (AU)

The use of teaching aids on the teaching-learning process of morphology and physiology subjects should be improved to strengthen knowledge of basic sciences in Medical Studies. The objective was to assess the use of teaching aids on the teaching-learning process of morphology and physiology of the osteomyoarticular system on 1st academic year of medical studies at "Dr. Ernesto Guevara de la Serna" Medical School. Pinar del Rio. Empirical and theoretical methods were used along with descriptive statistics procedures. Professors of the subjects were taken as a target group (n=6), as well as the consultants of the subject (n=4) and students of the 1st academic year of medical studies receiving the subject of Human Morphology and Physiology II (n=161) during 2010-2011. No correspondence was found regarding the organization of contents on CDs and the ones currently included in the syllabus of the subject, criteria of experts, professors and students in relation to the difficulties with the teaching aids, which was verified in the majority of the responses. The teaching aids used to teach the morphology and physiology of osteomyoarticular system give no responses to an interactive learning, where knowledge must be built and for the students to develop their own learning strategies, that is why it is necessary to better the teaching aids (AU)

Transgenic overexpression of insulin receptor substrate 1 in hepatocytes enhances hepatocellular proliferation in young mice only.

AIM: The insulin receptor substrate-1 (IRS-1) is a multisite docking protein which plays a central role in the signal transduction of growth factors such as insulin and insulin-like growth factors (IGF-1 and IGF-2). It is found to be frequently overexpressed in human hepatocellular carcinoma (HCC). METHODS: To study IRS-1 overexpression in hepatocytes in vivo, transgenic mice overexpressing IRS-1 exclusively in hepatocytes were created, showing enhanced hepatocyte proliferation in young animals. In the present study, the phenotype of IRS-1 transgenic animals was characterized over a period of two years. The livers of transgenic and control mice were analyzed for IRS-1 expression and phosphorylation, activation of the downstream mitogen-activated protein kinase (MAPK) cascade and phosphatidylinositol 3' kinase (PI3'K) and macroscopical and histological abnormalities. RESULTS: The enhanced hepatocyte proliferation observed in young IRS-1 transgenic animals was no longer detectable in adult mice. Despite constitutive overexpression and phosphorylation of IRS-1, MAPK- and IRS-1-associated PI3'K activity were significantly reduced in older transgenic mice. Furthermore, no premalignant lesions or HCC were detected in IRS-1 transgenic animals up to the age of 24 months. CONCLUSIONS: Therefore, additional mechanisms such as enhanced growth factor expression or impaired negative feedback control mechanisms may augment IRS-1 overexpression in human hepatocarcinogenesis.

Immunotherapy of murine hepatocellular carcinoma by alpha-fetoprotein DNA vaccination combined with adenovirus-mediated chemokine and cytokine expression.

Most hepatocellular carcinomas (HCCs) express oncofetal alpha-fetoprotein (AFP). We and others have demonstrated efficient tumor control mediated by cellular immune responses in mice bearing subcutaneous tumors derived from the AFP-expressing murine HCC cell line Hepa 1-6 by DNA vaccination against AFP. In the present study, we examined AFP DNA vaccination in the AFP-expressing primary murine HCC model BW7756. In this model AFP DNA vaccination resulted in only minimal lymphocytic infiltration and failed to control tumor growth. To augment the AFP-specific cellular immune response, intratumoral expression of chemokine IP-10 (interferon-inducible protein-10) and the proinflammatory cytokine interleukin (IL)-12 by adenoviral vectors (AdmIL-12 and AdmIP-10) was analyzed. Intratumoral injection of AdmIL-12 and AdmIP-10 resulted in transient tumor regressions, without prolongation of animal survival. By contrast, AFP DNA vaccination followed by intratumoral injection of AdmIL-12 and AdmIP-10 resulted in tumor regression in all animals and in prolongation of animal survival; in 25% of animals the tumors became undetectable. This study demonstrates for the first time that activation of effector cells against a tumor antigen induced by the combination of DNA vaccination and intratumoral chemokine and cytokine expression is superior to the respective treatment strategies alone. This effect may be mediated by attraction of activated effector cells to the tumor tissue.

The fraction of perforin-expressing HIV-specific CD8 T cells is a marker for disease progression in HIV infection.

OBJECTIVE: Perforin is an important component of the death machinery of cytotoxic T cells (CTL). To evaluate functional differences between HIV- and cytomegalovirus (CMV)-specific CTL of coinfected patients, the frequencies of the respective perforin-expressing T cells were analysed in a rapid whole blood assay. METHODS: Whole blood of HIV- and CMV-infected individuals was specifically stimulated by HIV-1 Pr55(gag) or complete CMV antigen, and activation-induced intracellular cytokine and perforin expression in CD8 T cells was analysed by flow cytometry. RESULTS: Perforin-expressing HIV-1- and CMV-specific CD8 T cells can be quantified simultaneously. Within a patient, the frequency of such HIV-specific CD8 T cells in peripheral blood was lower than the frequency of the respective CMV-specific cells. The number of the perforin-expressing HIV-specific CD8 T cells inversely correlated with the peripheral blood CD4 T cell count. CONCLUSIONS: The differential fractions of perforin-expressing virus-specific CD8 T cells in HIV and CMV double infection might be caused by differences in priming and trafficking to or from replication sites. However, without knowing the underlying mechanism, the fraction of perforin-expressing HIV-specific CD8 T cells provides another surrogate marker for disease progression.