Structural, immunohistochemical and molecular features of placentas and placental sites after in vitro fertilization with donor eggs (surrogate motherhood).

OBJECTIVE: to identify structural, immunohistochemical and molecular features of placentas and placental sites afterin vitro fertilization (IVF) with donor eggs (surrogate motherhood). STUDY DESIGN: morphological and immunohistochemical studies were performed on placental material obtained after delivery by caesarean section. The study included 26 women patients whose pregnancy resulted from IVF with a donor egg (IVF-SM group). The comparison group included 13 women patients whose pregnancy occurred after IVF with their own eggs (IVF-OE). Immunohistochemistry of biopsy material was performed using mouse antibodies to total cytokeratin (clone AE1/AE3) and murine antibodies to HLA-DR (clone TAL.1B5). Molecular studies were performed on DNA samples isolated from venous blood. HLA-DNA-TEH reagent kits and polymerase chain reaction were used for genotyping the main human histocompatibility complex class II (DQA1, DQB1 and DRB1). RESULTS: Histological examination of placenta in IVF-SM group showed a high incidence of central ischemic infarctions (69% of cases), dissociated cotyledon development (61%), pathological villus immaturity (46%) and massive perivillous fibrin deposition (73%). This group also had a pronounced lymphoplasmacytic deciduitis, which was 2 times higher than in the control group, and an expressed inflammatory process in the placental sites. Remodeling of the spiral arteries was incomplete in more than 40% of cases, and 30% of spiral arteries had no gestational changes. In comparison group, a complete gestational adjustment was found in more than 90% of spiral arteries. A focal lymphohistiocytic infiltration in perivascular regions, and a decrease in the number of multinucleated cells as compared with the control were also observed. For seven female surrogate mothers and their children, allelic polymorphisms of genes of HLA II class were studied. CONCLUSION: Placental material of women from IVF-SM group is characterized by complex immune response in sites of tight contact between maternal and fetal tissues. The immune pathogenesis is associated with an increase in the number of HLA-DR positive cells, defects in remodeling of the spiral arteries, development of areas of chronic inflammation in perivascular regions, and a decrease in the number of multinucleated cells. Genetic incompatibility between alleles of HLA II genes can be a molecular predictor of impaired immune tolerance.

[Myocarditis and fibroplastic endocarditis in Wegener's granulomatosis].

The paper describes an autopsy case of Wegener's granulomatosis with a rare variant of a cardiac lesion--the concurrence of aortic valvular disease (aortic regurgitation), fibroplastic endocarditis, and complete transverse blockade, followed by pacemaker implantation. The direct cause of death in the female patient was multiple organ dysfunction: progressive chronic heart failure, adult respiratory distress syndrome, acute hepatorenal failure, and pancreonecrosis. The data available in the Russian and foreign literature on investigations dealing with cardiac pathology in patients with Wegener's granulomatosis are analyzed.

Differential regulation of synapsin phosphorylation by monocular deprivation in juveniles and adults.

The rodent visual cortex retains significant ocular dominance plasticity beyond the traditional postnatal critical period. However, the intracellular mechanisms that underlie the cortical response to monocular deprivation are predicted to be different in juveniles and adults. Here we show monocular deprivation in adult, but not juvenile rats, induced an increase in the phosphorylation of the prominent presynaptic effecter protein synapsin at two key sites known to regulate synapsin function. Monocular deprivation in adults induced an increase in synapsin phosphorylation at the PKA consensus site (site 1) and the CaMKII consensus site (site 3) in the visual cortex ipsilateral to the deprived eye, which is dominated by non-deprived eye input. The increase in synapsin phosphorylation was observed in total cortical homogenate, but not synaptoneurosomes, suggesting that the pool of synapsin targeted by monocular deprivation in adults does not co-fractionate with excitatory synapses. Phosphorylation of sites 1 and 3 stimulates the release of synaptic vesicles from a reserve pool and increases in the probability of evoked neurotransmitter release, which may contribute to the strengthening of the non-deprived input characteristic of ocular dominance plasticity in adults.

[The comparison of laparoscopic and open lymphodissection for gastric cancer].

Lymphodissection for gastric cancer was performed in 174 patients. Among 116 patients, operated traditionally, 58% had cancer stages I and II. 58 patients operated laparoscopically revealed cancer stages I and II in 55.1%. The mean number of removed lymph nodes was 28.9+/-1.2 and was higher after laparoscopic operation (p=0.652). The duration of the laparoscopic lymphodissection operation was slightly shorter. Blood loss was significantly less during the laparoscopic procedure. The mean survival time for patients operated laparoscopically and traditionally was 30 and 26.6 months respectively.

Dissociaton of conditioned eye and limb responses in the cerebellar interpositus.

Thompson and colleagues have demonstrated that the lateral interpositus nucleus of the cerebellum is the essential locus for the classical conditioning of the somatic eyeblink response. Preliminary studies reported that lesioning the cerebellar interpositus nucleus ipsilateral to the side of training also appears to abolish conditioned limb flexion responses. Previous studies have suggested that the interpositus nucleus is somatotopically organized with the eye being represented laterally and the hindlimb medially. Presently, we employed a double dissociation paradigm to examine the effects of muscimol (a GABA(A) agonist) injections on eyeblink versus limb flexion conditioned responses in the ipsilateral cerebellar interpositus nucleus of New Zealand white rabbits. For eyeblink conditioning, the conditioned stimulus (CS) was a 14-V lamp bulb and the unconditioned stimulus (US) was a 3-psi corneal airpuff to the left eye. For limb flexion conditioning, the CS was a 1-kHz, 85-95 dB SPL tone and the US was a 3- to 5-mA shock to the upper left hindlimb. Upon training on both responses to a 60-100% criterion, the rabbits were then tested on eyeblink and limb flexion responses after injections of muscimol (0.1-0.3 mul of a 0.01- to 1.0-M solution) into either the lateral (eyeblink) or medial (limb flexion) interpositus nucleus. We have been able to successfully decrease or abolish the percent conditioned responses (CRs) of both the eyeblink and limb flexion conditioning selectively without affecting the other. These results thus lend further support for the notion of the existence of a somatotopic map in the interpositus nucleus for learning.

Comparative analysis of CD1a, S-100, CD83, and CD11c human dendritic cells in normal, premalignant, and malignant tissues.

A number of antibodies that recognize human dendritic cells (DC) have been identified. The main aim of this study was to compare and contrast different antigen retrieval techniques using both enzymatic and non-enzymatic treatments in order to determine the expression and distribution of several DC markers on formalin-fixed, paraffin-embedded tissues. Normal human lung, oral epithelial hyperplasia lesions, oral squamous cell carcinoma, and prostate adenocarcinoma tissues were evaluated using a panel of DC specific antibodies. The results of immunohistochemical staining for CD83, CD1a, CD11c, and S-100 DC markers were compared following the different antigen retrieval approaches. The overall best results for the analysis of tumor-associated DC were obtained with the enzymatic methods. Protease XXIV digestion was determined to be essential for detection of S-100 and CD11c positive DC, whereas trypsin and pepsin were required for the recognition of CD1a and CD83 expressing tumor-associated DC. These results could be easily adapted for routine practice and should be useful for characterization of the DC system in cancer patients for both diagnostic and prognostic purposes. In addition, standardized procedures for evaluating different subpopulations of tumor-associated DC should bring new insights in understanding of DC-tumor cell interaction.

Estenosis de uretra anterior complejas. Resolución quirúrgica / Complex anterior urethral strictures. Surgical treatment

Objetivo: Evaluar la respuesta terapéutica quirúrgica en pacientes con estenosis de uretra bulbar(EUB) adyacentes al piso perineal, coincidentemente con una estenosis de la uretra péndula(EUP). Material y métodos: Se estudió una población de 15 pacientes con estas características. Se definieron dos grupos según la técnica empleada en la resoluciónde la EUB: Grupo a) 13 pacientes requirieron la realización de una Resección Anastomosis Primaria (RAP). Grupo b) en 2 casos se emplearon técnicas en dos tiempos. La EUP fue resuelta con un colgajo fasciocutáneo longitudinal de piel de pene a lo Orandi o transversal de prepucio. El seguimiento promedio de toda la población fue 4,3 años. Resultados: El 33,33 por ciento de los pacientes tuvieron que ser reintervenidos quirúrgicamente: Grupo a) fueron reintervenidos el 23 por ciento y en el grupo b) el 100 por ciento. Topografía de las recurrencias: Grupo a) en todos los casos involucraba la EUB, Grupo b) se registraron estenosis a nivel de la EUB en un caso y de la boca proximal del Orandi en otro. La plástica que más recurrencia registró fue la RAP. Los tiempos de recurrencia fueron bajos con una media de 3,48 meses (2 a 9 meses). Conclusión: a) La recurrencia observada fue elevada, especialmente en las RAP. b) La recurrencia suele ser temprana y suele requerir resolución quirúrgica, siendo improbable que opciones mínimamente invasivas sean la opción a considerar

Estenosis de uretra anterior complejas. Resolución quirúrgica / Complex anterior urethral strictures. Surgical treatment

Objetivo: Evaluar la respuesta terapéutica quirúrgica en pacientes con estenosis de uretra bulbar(EUB) adyacentes al piso perineal, coincidentemente con una estenosis de la uretra péndula(EUP). Material y métodos: Se estudió una población de 15 pacientes con estas características. Se definieron dos grupos según la técnica empleada en la resoluciónde la EUB: Grupo a) 13 pacientes requirieron la realización de una Resección Anastomosis Primaria (RAP). Grupo b) en 2 casos se emplearon técnicas en dos tiempos. La EUP fue resuelta con un colgajo fasciocutáneo longitudinal de piel de pene a lo Orandi o transversal de prepucio. El seguimiento promedio de toda la población fue 4,3 años. Resultados: El 33,33 por ciento de los pacientes tuvieron que ser reintervenidos quirúrgicamente: Grupo a) fueron reintervenidos el 23 por ciento y en el grupo b) el 100 por ciento. Topografía de las recurrencias: Grupo a) en todos los casos involucraba la EUB, Grupo b) se registraron estenosis a nivel de la EUB en un caso y de la boca proximal del Orandi en otro. La plástica que más recurrencia registró fue la RAP. Los tiempos de recurrencia fueron bajos con una media de 3,48 meses (2 a 9 meses). Conclusión: a) La recurrencia observada fue elevada, especialmente en las RAP. b) La recurrencia suele ser temprana y suele requerir resolución quirúrgica, siendo improbable que opciones mínimamente invasivas sean la opción a considerar(AU)

Uretroplastia con injerto de mucosa bucal en el tratamiento de la estenosis de uretra anterior / Urethroplasty with buccal mucosa graft in the treatment of anterior urethral stricture

Introducción: Si bien los colgajos de piel genital utilizados para el tratamiento de la estenosis de uretra anterior han probado su efectividad a lo largo del tiempo, el empleo de injertos de mucosa bucal, ha reclutado nuevos adeptos, con resultados comparables. Material y método: Fueron tratados 33 pacientes con injertos de mucosa bucal con un seguimiento promedio de 14,7 meses. La estenosis se localizaba en la uretra bulbar (EUB) en 26 casos, en la uretra péndula (EUP) en 6 pacientes y presentaba extensión a uretra bulbar y peneana (EUBP) en 1 caso. El injerto de mucosa bucal se utilizó en forma de Anastomosis ampliada o Ampliación uretral de acuerdo con la topografía de la estenosis y al grado de espongiofbrosis. El lecho donante lo constituyó la cara bucal del carrillo. Los defectos uretrales a tratar oscilaron entre los 3 y 7 cm de longitud. Resultados: Si se toma en cuenta la recurrencia en el sitio de la plástica original la recurrencia observada fue 3/33 (9,1 por ciento). La incidencia de recurrencia obtenida, en el corto plazo, es comparable a otras series que describen el uso de injertos de mucosa bucal y a la recurrencia observada en colgajos de piel genital, en el corto plazo.

Uretroplastia con injerto de mucosa bucal en el tratamiento de la estenosis de uretra anterior / Urethroplasty with buccal mucosa graft in the treatment of anterior urethral stricture

Introducción: Si bien los colgajos de piel genital utilizados para el tratamiento de la estenosis de uretra anterior han probado su efectividad a lo largo del tiempo, el empleo de injertos de mucosa bucal, ha reclutado nuevos adeptos, con resultados comparables. Material y método: Fueron tratados 33 pacientes con injertos de mucosa bucal con un seguimiento promedio de 14,7 meses. La estenosis se localizaba en la uretra bulbar (EUB) en 26 casos, en la uretra péndula (EUP) en 6 pacientes y presentaba extensión a uretra bulbar y peneana (EUBP) en 1 caso. El injerto de mucosa bucal se utilizó en forma de Anastomosis ampliada o Ampliación uretral de acuerdo con la topografía de la estenosis y al grado de espongiofbrosis. El lecho donante lo constituyó la cara bucal del carrillo. Los defectos uretrales a tratar oscilaron entre los 3 y 7 cm de longitud. Resultados: Si se toma en cuenta la recurrencia en el sitio de la plástica original la recurrencia observada fue 3/33 (9,1 por ciento). La incidencia de recurrencia obtenida, en el corto plazo, es comparable a otras series que describen el uso de injertos de mucosa bucal y a la recurrencia observada en colgajos de piel genital, en el corto plazo. (AU)

Transcriptional activation of TRADD mediates p53-independent radiation-induced apoptosis of glioma cells.

Survival of patients with Glioblastoma Multiforme (GM), a highly malignant brain tumor, remains poor despite concerted efforts to improve therapy. The median survival of patients with GM has remained approximately 1 year regardless of the therapeutic approach. Since radiation therapy is the most effective adjuvant therapy for GM and nearly half of GM tumors harbor p53 mutations, we sought to identify genes that mediate p53-independent apoptosis of GM cells in response to ionizing radiation. Using broad-scale gene expression analysis we found that following radiation treatment, TRADD expression was induced in a uniquely radiosensitive GM cell line but not in radioresistant GM cell lines. TRADD over-expression killed GM cells and activated NF-kappa B. We found that blocking the TRADD-mediated pathway using a dominant-negative mutant of FADD (FADD-DN) enhanced radiation resistance of GM cells, as reflected in both susceptibility to apoptosis and clonogenic survival following irradiation. Conversely, stable expression of exogenous TRADD enhanced radiation-induced apoptosis of GM cell lines, reflecting the biological significance of TRADD regulation in p53-independent apoptosis. These findings generate interest in utilizing TRADD in gene therapy for GM tumors, particularly in light of its dual function of directly inducing rapid apoptosis and sensitizing GM cells to standard anti-neoplastic therapy.

p27Kip1 is required for PTEN-induced G1 growth arrest.

The tumor suppressor PTEN is one of the most commonly inactivated genes in human cancer. Glioblastoma multiforme cells harboring mutant PTEN have abnormally high levels of 3' phosphoinositides and elevated protein kinase B activity. Expression of wild-type PTEN in glioma cells, containing endogenous mutant PTEN, reduces 3' phosphoinositides levels, inhibits PKB activity, and induces G1 cell cycle arrest. We investigated the mechanism of the PTEN-induced growth arrest in glioma cell lines. Expression of PTEN is associated with increased expression of p27Kip1, decreased expression of cyclins A and D3, inhibition of cdk2 activity, and dephosphorylation of pRb. Inactivation of p53, by the human papilloma virus E6 oncoprotein, does not prevent PTEN-induced G1 arrest, implying that p53 is not required for G1 arrest. In contrast, p27Kip1 antisense oligonucleotides abrogated the growth arrest induced by PTEN. Furthermore, blocking p27Kip1 expression prevented the PTEN-induced reduction of cyclin-dependent kinase 2 activity, indicating that p27Kip1 functions upstream of cyclin-dependent kinase 2 in the PTEN regulatory cascade. These results implicate p27Kip1 as a critical mediator of PTEN-induced G1 arrest.

Functional and clinical outcomes of limb-sparing therapy for pediatric extremity sarcomas.

PURPOSE: To determine the clinical and functional outcomes of children undergoing limb-sparing therapy for extremity sarcomas. METHODS AND MATERIALS: We retrospectively reviewed 30 patients, age < or = 21 years, who were treated between l979 and l998 with external beam radiotherapy as a component of limb-sparing therapy for primary sarcomas of the extremity at UCSF. Included were patients for whom complete follow-up and functional outcome assessments were available. We assessed the patterns of failure, overall survival, disease-free survival, local control, and limb function. RESULTS: At a median follow-up of 3 years, 12 of the 30 patients recurred: 3 locally, 8 distantly, and 1 with synchronous local and distant disease as site of first progression. Eighteen patients were alive with no evidence of disease. The median overall survival was 10 years, with a median disease-free survival of 8 years. Functional outcome assessment revealed 15 patients retained excellent, 12 good, 1 fair, and 2 poor limb function. CONCLUSION: In pediatric patients receiving limb-sparing therapy, 90% maintained excellent or good limb function without compromising survival, demonstrating the validity of limb preservation in children with extremity sarcomas.

The role of magnetic resonance cholangiography in the management of children and young adults after liver transplantation.

We reviewed the results of 50 magnetic resonance (MR) cholangiograms to evaluate their usefulness in directing clinical management in young patients after liver transplantation (LTx). Thirty-two patients underwent 50 MR cholangiograms on a 1.5-T unit. Studies were performed from 1 week to 16 yr after LTx. Indications included biochemical abnormalities with (n = 19) or without (n = 16) biopsy evidence for chronic rejection, sepsis (n = 14), and intractable ascites (n = 1). Original interpretations were compared to laboratory and ultrasound findings, and clinical outcome. Of 19 studies performed on 14 patients with biopsy evidence of chronic rejection, 16 were abnormal on MR (but only one was abnormal on ultrasound), resulting in corrective surgery (n = 1), re-Tx (n = 1), and endoscopic dilatation (n = 1). Of 16 studies on 16 patients with biochemical abnormalities without evidence of chronic rejection on biopsy, 14 were abnormal on MR (but only five of 13 on ultrasound), leading to corrective surgery (n = 3) and re-listing for Tx (n = 3). Thirteen of 14 studies on six patients with sepsis were abnormal on MR (five of nine were abnormal on ultrasound), identifying surgically correctable strictures (n = 2), and leading to re-Tx (n = 1) and percutaneous biliary drainage procedures (n = 2). The one patient with ascites had a normal study. We advocate usage of MR cholangiography for the detection of biliary complications after LTx, particularly in those patients who present with biochemical abnormalities that are not easily explained by acute cellular rejection or viral infection and in those with biliary sepsis.

Intraoperative radiation therapy for high-risk pediatric neuroblastoma.

PURPOSE: To evaluate the efficacy of intraoperative radiation therapy (IORT) in the treatment of high-risk pediatric neuroblastoma. METHODS AND MATERIALS: Between 1986 and 1998, 23 children received IORT for pediatric neuroblastoma. Electron beam energies ranged from 4 MeV to 16 MeV and median dose was 10 Gy (7-16 Gy). RESULTS: Twenty-one of 23 patients were classified as high-risk. A gross total resection (GTR) was achieved in 18 patients, of whom 6 experienced disease recurrence, 2 of which included a locoregional relapse as a component of failure. Fourteen of 18 patients receiving IORT after a GTR are disease-free survivors. A second subset of 5 patients had a subtotal resection (STR), with gross residual disease remaining after surgery. All 5 patients recurred locally, and all died of their disease. IORT was extremely well-tolerated in our cohort. Surgical resection and IORT resulted in the narrowing of the abdominal aorta and an atrophic kidney in 1 patient. CONCLUSIONS: For high-risk neuroblastoma patients, IORT as the only radiotherapy to the primary, produced excellent local control after a GTR. However, IORT as the sole radiotherapy to the primary was inadequate for patients with extensive adenopathy or an STR. In this setting, we are exploring the use of IORT as a boost in conjunction with external beam radiation therapy.

Loss of PTEN facilitates HIF-1-mediated gene expression.

In glioblastoma-derived cell lines, PTEN does not significantly alter apoptotic sensitivity or cause complete inhibition of DNA synthesis. However, in these cell lines PTEN regulates hypoxia- and IGF-1-induced angiogenic gene expression by regulating Akt activation of HIF-1 activity. Restoration of wild-type PTEN to glioblastoma cell lines lacking functional PTEN ablates hypoxia and IGF-1 induction of HIF-1-regulated genes. In addition, Akt activation leads to HIF-1alpha stabilization, whereas PTEN attenuates hypoxia-mediated HIF-1alpha stabilization. We propose that loss of PTEN during malignant progression contributes to tumor expansion through the deregulation of Akt activity and HIF-1-regulated gene expression.

Overexpression of E2F1 in glioma-derived cell lines induces a p53-independent apoptosis that is further enhanced by ionizing radiation.

Glioma cell lines show variable responses to radiation in a manner influenced by their p53 status. Irradiation of glioma cell lines does not generally induce apoptosis. When wild-type p53 is present, these cells undergo a G1 arrest that is closely associated with increased radiosensitivity as measured by clonogenic survival. Previously, others have shown that dysregulated overexpression of E2F1 induces apoptosis in cell lines with either functional or inactivated p53. We found that regardless of p53 status, apoptosis induced by overexpression of E2F1 in glioma cell lines was further enhanced by treatment with ionizing radiation. BAX induction did not follow E2F1 overexpression or irradiation in the glioma cell lines tested. Thus, the apoptotic response of glioma-derived cells to irradiation can be enhanced by E2F1 by a mechanism that does not involve the induction of BAX.

MR cholangiography in children and young adults with biliary disease.

OBJECTIVE: Our objective was to describe the MR cholangiography findings for young patients with suspected biliary disease who underwent half-Fourier acquisition fast spin-echo technique with respiratory triggering. SUBJECTS AND METHODS: Twenty-eight MR cholangiography studies were performed in 22 patients on a 1.5-T MR unit. Ten of these 22 patients had undergone liver transplantation. RESULTS: MR cholangiography revealed abnormalities of both the extrahepatic and the intrahepatic major and minor bile duct systems, despite the small diameter of the duct system in this group of patients. Four patterns of biliary disease were shown: global dilatation of extrahepatic or intrahepatic ducts (n = 7); segmental, uniform dilatation of central or peripheral intrahepatic ducts (n = 9); segmental, nonuniform dilatation of central or peripheral intrahepatic ducts (n = 2); and fusiform ectasia with segmental, irregular intrahepatic dilatation and bile lakes (n = 2). The findings of eight studies were interpreted as normal. The four patterns of abnormalities were correlated with the results from percutaneous transhepatic cholangiography, T-tube cholangiography, and liver biopsy and with clinical and surgical information, as available. CONCLUSION: MR cholangiography is a noninvasive technique for evaluation of biliary disease. The improved resolution afforded by respiratory triggering permits evaluation of both major and minor bile ducts, even in young, uncooperative subjects. Four patterns of abnormalities were prospectively identified, correlated with other information, and used to direct clinical treatment.