Differentiation and secretory activities of cultured human placental cytotrophoblast.

Ultrastructural, autoradiographic, immunofluorescent and biochemical techniques were used to characterize primary cultures of term placental cytotrophoblast in order to gain insight into the differentiation and secretory capacities of the cellular component of human trophoblast. Trypsin treatment of placental villi allowed isolation of a predominantly cytotrophoblast cell population that maintained viability up to 13 weeks in monolayer culture. Autoradiographic studies of tritiated thymidine incorporation identified a smaller diameter mononucleated cell population that was mitotically active and developed into larger diameter mononucleated cells and into multinucleated cells during culture. Ultrastructurally, cultured cells formed desmosomes, had an extensive network of cytoplasmic microfilaments and contained the organelles for hormone synthesis and secretion. These cells secreted steroid hormones, secreted Schwangerschafts protein I, actively incorporated tritiated glycoprotein precursors and expressed surface immunoreactivity for the beta-subunit of human chorionic gonadotrophin (hCG). However, medium concentrations of hCG and human placental lactogen dropped rapidly to undetectable levels after 14 days in primary culture. Cells grown beyond confluence differentiated into 1 to 2 mm structures with a villus-like histology. Our studies indicate that cytotrophoblast can secrete steroids, cytotrophoblast differentiation occurs in vitro in the absence of maternal tissues, hCG synthesis occurs in cultured cytotrophoblast and medium concentrations of placental protein hormones are not the best indicators of cell viability for cultures of cytotrophoblast.

Active immunity and T-cell populations in pigs intraperitoneally inoculated with baculovirus-expressed transmissible gastroenteritis virus structural proteins.

The intraperitoneal inoculation of pigs with baculovirus-expressed transmissible gastroenteritis virus (TGEV) structural proteins (S, N, M) in conjunction with thermolabile Escherichia coli mutant toxin (LT-R192G) in incomplete Freund's adjuvant (IFA) was tested in an attempt to elicit active immunity to TGEV in gut-associated lymphoid tissues (GALT). Four groups of 63 (1-5-week-old) suckling, TGEV-seronegative pigs were used to assess the efficacy of the recombinant protein vaccine (group 3) in comparison with sham (group 1), commercial vaccine (group 2), and virulent TGEV Miller-strain-inoculated pigs (group 4). The TGEV-specific mucosal and systemic immune responses were measured after in vivo and in vitro stimulation with TGEV-antigens. The major T-cell subset distribution was analyzed in vivo and in vitro after stimulation of mononuclear cells with TGEV (from mesenteric lymph nodes of group 3 inoculated with TGEV-recombinant proteins). Induction of active immunity was assessed by challenge of pigs with virulent TGEV at 27 days of age. Baculovirus-expressed TGEV proteins coadministered with LT-R192G in IFA induced mesenteric lymph node immune responses associated with IgA-antibodies to TGEV and partial protection against TGEV-challenge. The high titers of serum IgG- and virus-neutralizing-antibodies to TGEV in group 3 pigs most likely reflected the dose of TGEV S-protein administered. At the day of TGEV-challenge, the in vitro stimulation of mononuclear cells from the mesenteric lymph nodes of group 3 pigs with inactivated TGEV resulted in an increase in double positive (CD4+CD8+), natural killer (CD2+CD4-CD8+dim) and cytotoxic (CD2+CD4-CD8+bright) T-cell phenotypes, accompanied by increased expression of interleukin-2 receptor and a decrease of the null (CD2-CD4-CD8-/SW6+) cell phenotype.

The influence of portoenterostomy with stoma on morbidity in pediatric patients with biliary atresia undergoing orthotopic liver transplantation.

A portoenterostomy (PE) procedure for extrahepatic biliary atresia (EHBA) is sometimes performed with a stoma in an attempt to reduce the incidence of acute cholangitis. The purpose of this study was to determine if the presence of a stoma increased the complication rate of patients undergoing orthotopic liver transplantation (OLT) for EHBA. The medical records of 42 consecutive patients with EHBA who underwent primary OLT between October 1988 and October 1991 were retrospectively reviewed. Three patients were excluded, since their grafts were lost within 3 days of OLT. The remaining 39 patients were divided into three groups: no PE (n = 7), PE without stoma (n = 23), and PE with stoma (n = 9). The mean age of the whole group was 19.62 +/- 24.37 months, with a range of 5 to 132 months. Mean weight was 9.62 kg, with a range of 4.2 to 41 kg. Survival at 3 and 12 months as well as number of retransplantations were similar among the three groups. However, at the time of OLT increased morbidity was observed, consisting of increased operative time and number of reoperations, whether or not the stoma had been closed prior to OLT.

Identification of cultured pleural fluid cells by a direct immunofluorescence technique.

A preliminary study demonstrated intracytoplasmic polyvalent immunoglobulin in human pleural fluid cells using a direct immunofluorescence technique. Of 33 pleural-fluid cell samples tested, 31 contained cells that initially stained positive for polyvalent immunoglobulin. The two negative specimens were from malignant effusions. Eleven of the specimens were cultured for seven days; nine of the eleven cultured specimens tested negative for polyvalent immunoglobulin after four days in culture. The remaining two specimens showed a marked decrease in positivity. After adding cell-free pleural fluid to the cultures on day four, the nine negative samples became positive again by day seven; the two slightly positive cases became strongly positive. The significance of these findings is discussed.

Laparoscopically guided blood salvage and autotransfusion in splenic trauma: a case report.

Salvage of intraperitoneal blood with autotransfusion is a well-accepted practice. Laparoscopic examination is gaining popularity and holds diagnostic promise for the evaluation of trauma patients. We describe herein the successful combination of these techniques in a patient who sustained blunt abdominal trauma, facilitating splenic salvage, autotransfusion, and avoidance of laparotomy.

Comparison of a lesion-inducing isolate and a non-lesional isolate of Candida albicans in an immunosuppressed rat model of oral candidiasis.

Two distinct strain-related patterns of organism-host interaction on dorsal tongue of immunocompetent rats have been identified for Candida albicans: some isolates induce mucosal lesions, while other isolates penetrate the keratin layer but do not produce a lesion. This study examined the behavior of each of the two types of isolates in a cyclosporin-immunosuppressed rat model. Groups B (normal) and D (cyclosporin) were orally inoculated with a lesion-inducing isolate of C. albicans, while a non-lesional isolate was given to Groups A (normal) and C (cyclosporin). A typical dorsal tongue lesion developed in 4/18 rats in Group B and in 13/16 in Group D (P = 0.00267). No significant difference in infection rate between the normal and cyclosporin-treated animals was seen for the non-lesional isolate. The lack of a host inflammatory response associated with the non-lesional isolate may represent an ecologic advantage for the organism.