Preservation of splenic immune functions by female sex hormones after trauma-hemorrhage

OBJECTIVES: Immune functions are markedly depressed in males but not in proestrus females after trauma-hemorrhage. Nonetheless, it is unclear what role sex steroids play in the maintenance of immune function in females after trauma-hemorrhage. DESIGN: Prospective, controlled animal study. SETTING: University research laboratory. SUBJECTS: Eight-week-old female CBA/J mice. INTERVENTIONS: Mice underwent sham-ovariectomy or ovariectomy. Two weeks thereafter, ovariectomized and proestrus sham-ovariectomized mice were subjected to laparotomy (i.e., soft tissue trauma) and hemorrhagic shock (35 +/- 5 mm Hg for 90 mins, resuscitated) or sham operation. Splenocyte proliferation and interleukin-2, interleukin-3, and interferon-gamma release were determined at 2 hrs after trauma-hemorrhage. MEASUREMENTS AND MAIN RESULTS: These immune functional capacities were maintained in proestrus sham-ovariectomized mice after trauma-hemorrhage, whereas they were suppressed in ovariectomized mice subjected to trauma-hemorrhage. 17beta-Estradiol in vitro had no effect on splenocyte functions in proestrus sham-ovariectomized females; however, addition of 17beta-estradiol to splenocytes from ovariectomized females subjected to trauma-hemorrhage normalized immune functional capacities. CONCLUSIONS: These findings suggest that elevated circulating 17beta-estradiol in proestrus females plays a direct role in the maintenance of immunocompetence after trauma-hemorrhage.     

Distinct ongoing Ig heavy chain rearrangement processes in childhood B- precursor acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) is thought to arise from the clonal expansion of a single transformed precursor cell. However, an oligoclonal Ig heavy chain (IgH) rearrangement pattern has been observed in 30% of ALL patients and was shown to be the result of ongoing rearrangement events. The extent and nature of these ongoing rearrangement processes in individual patients has so far remained obscure. We performed a detailed analysis of leukemic VHDJH rearrangements in three children with B-precursor ALL at diagnosis and one B-lymphoid blast crisis of a child with Ph+ chronic myeloid leukemia at diagnosis and relapse. The children were selected because they presented with multiple IgH rearrangements on Southern blot analysis. Polymerase chain reaction analysis of leukemic cells from two B-precursor ALL patients showed exclusively two groups of related sequences resulting from VH gene replacement events. Most VH gene replacements involved 3' located acceptor VH genes. Analysis of cells from the other B-precursor ALL patient showed exclusively related sequences as a result of VH gene joinings to a pre-existing DJH rearrangement. In the B-lymphoid blast crisis, a single germline precursor cell had generated multiple unrelated rearrangements and additional groups of related rearrangements resulting from VH to DJH joinings. Direct proof for the VH to DJH joining mechanism was obtained by amplification of the expected preexisting DJH rearrangements. Our findings suggest that the pattern of ongoing rearrangements in an individual patient reflects the IgH rearrangement status of the precursor cell at the time of malignant transformation. Sequence analysis of VHDJH rearrangements at diagnosis may therefore allow a prediction of the reliability of complementarity determining region 3 probes for the detection of minimal residual disease.     

Cellular and molecular contributions of the cardiac neural crest to cardiovascular development

Normal development of the heart and great arteries requires participation of the cardiac neural crest. Ectomesenchymal cells from this area of the neural crest migrate to pharyngeal arches 3, 4, and 6, where they support development of the aortic arch arteries. Cells continue migration from the pharyngeal arches to specific sites in the outflow tract. Removal of the neural crest results in two types of malformations: outflow septation defects and alignment defects. The genesis of these two types of defects is by two different mechanisms. Outflow septation is disturbed when a critical number of cells does not reach the outflow tract. Alignment is altered by an as yet unknown secondary mechanism that is transmitted upstream to the heart from the pharyngeal arches. Aortic arch artery and ventricular development as well as hemodynamic parameters are abnormal from an early age. Some possible molecular mechanisms involved in specifying neural crest for participation in heart development are discussed.     

Rapid Rh D genotyping by polymerase chain reaction-based amplification of DNA

Rh (rhesus) D is the dominant antigen of the Rh blood group system. Recent advances in characterization of the nucleotide sequence of the cDNA(s) encoding the Rh D polypeptide allow the determination of the Rh D genotype at the DNA level. This can be of help in cases in which red blood cells are not available for phenotyping, eg, when in concerns a fetus. We have tested three independent DNA typing methods based on the polymerase chain reaction (PCR) for their suitability to determine the Rh D genotype. DNA derived from peripheral blood mononuclear cells from 234 Rh-phenotyped healthy donors (178 Rh D positive and 56 Rh D negative) was used in the PCR. The Rh D genotypes, as determined with a method based on the allele-specific amplification of the 3' noncoding region of the Rh D gene described by Bennett et al (N Engl J Med 329:607, 1993), were not concordant with the serologically established phenotypes in all cases. We have encountered 5 discrepant results, ie, 3 false-positive and 2 false-negative (a father and child). Rh D genotyping with the second method was performed by PCR amplification of exon 7 of the D gene with allele-specific primers. In all donors phenotyped as D positive tested so far (n = 178), the results of molecular genotyping with this method were concordant with the serologic results, whereas a false-positive result was obtained in one of the D-negative donors (also false-positive in the first method). Complete agreement was found between genotypes determined in the third method, based on a 600-bp deletion in intron 4 of the Rh D gene described by Arce et al (Blood 82:651, 1993), and serologically determined phenotypes. The Rh blood group system is complex, and unknown polymorphisms at the DNA level are expected to exist. Therefore, although genotypes determined by the method of Arce et al were in agreement with serotypes, it cannot yet be regarded as the golden standard. More experience with this or other methods is still needed.     

A phase I trial of intrapleural recombinant human interferon α (rHuIFNα2b) in patients with malignant pleural effusions

The use of intrapleural sclerosing agents to control reaccumulation of pleural fluid in patients with malignant effusions has been widely investigated. A phase I trial of intrapleural recombinant human interferon (rHuIFN2b) was initiated to determine the toxicity and maximal tolerated dose in this group of patients. rHuIFN2b was instilled as a single dose following chest tube (15/16) or percutaneous (1/16) drainage of cytologically proven malignant effusions. Doses of rHuIFN2b were escalated from 25×10⁶ to 200×10⁶ U/m² in cohorts of three to four patients. Toxicity was mild to moderate, and included chills, fever and chest pain, and resembled that produced by systemic administration of rHuIFN2b. Dose-limiting toxicity occurred at 200×10⁶ U/m² and consisted of hepatic enzyme elevations and renal failure. Partial control of the effusions was noted in two patients, with two additional patients having stable disease. Phase II trials of rHuIFN2b should utilize up to 150×10⁶ U/m² for intrapleural instillation.Abbreviation IFN interferon - MPE malignant pleural effusions Supported in part by a grant from the Schering Corporation, Kenilworth, N.J.     

Respiratory burst oxidase activation can be dissociated from phosphatidylinositol bisphosphate degradation in a cell-free system from human neutrophils

Activation of the respiratory burst oxidase in cell-free preparations from 32P-labeled neutrophils was compared with changes in levels of radioactively labeled phosphoinositides in the same preparations. With membrane particles, treatment with sodium dodecyl sulfate (SDS) in the presence of cytosol led to activation of the oxidase without an alteration in levels of labeled phosphatidylinositol 4,5-bisphosphate (PIP2) or phosphatidylinositol 4-phosphate (PIP). Conversely, solubilization of the membrane particles with deoxycholate resulted in loss of nearly 98% of the radioactive PIP2 without activation of the oxidase. In this solubilized preparation, the oxidase could subsequently be fully activated by SDS in the presence of cytosol, even though the labeled PIP2 was almost totally depleted. Two PIP2-derived second messengers, diacylglycerol and inositol 1,4,5-trisphosphate, as well as the protein kinase C activator phorbol myristate acetate (PMA), failed to activate the oxidase. These results suggest that in a cell- free preparation from human neutrophils, detergent-mediated activation of the respiratory burst oxidase is independent of changes in the levels of phosphoinositides or phosphoinositide-derived second messengers.     

Advanced maternal age and fetal growth inhibition in triplets

OBJECTIVE: To determine whether advanced maternal age is associated withfetal growth inhibition in triplets. STUDY DESIGN: We conducted a retrospective cohort study on triplet live births in the United States from 1995 through 1998. The outcomes of fetal growth inhibition measured were low birth weight, very low birth weight, preterm birth, very preterm birth and smallnessfor gestational age. We generated adjusted ORs after taking into account intracluster correlations using the generalized estimating equation framework. RESULTS: As compared to women of younger maternal age (20-29), mature (30-39) and older women (> or =40 years) with triplet gestations tended to have a lower likelihood offetal growth inhibition. Mean birth weight and mean gestational age at delivery increased with increasing maternal age in a dose-dependent pattern (p for trend < 0.0001). As compared to triplets born to younger mothers, those of older women were less likely to have low birth weight (OR=0.51, 95% CI=0.37-0.69) or very low birth weight (OR = 0.58, 95% CI = 0.47-0.72) or to be preterm (OR = 0.39, 95% CI = 0.27-0.56) or very preterm (OR = 0.67, 95% CI = 0.55-0.80). The riskfor small-for-gestational-age infants was comparable. CONCLUSION: Older maternal age is associated with morefavorable triplet fetal growth parameters, although the exact mechanisms of this paradox remain poorly understood.     

Outcomes of Stage I/II vulvar cancer patients after negative superficial inguinal lymphadenectomy

OBJECTIVES: To investigate the patterns of recurrence associated with superficial inguinal lymphadenectomy (SupIL) and vulvectomy for patients with Stage I/II vulvar cancer. METHODS: A retrospective chart review identified patients from 1990-2001 with Stage I/II vulvar cancer that underwent SupIL and vulvectomy. Survival was analyzed using the Kaplan-Meier method with Fisher Exact and Chi-square tests for comparisons between groups. RESULTS: 65 patients with Stage I/II vulvar cancer with a pathologically negative SupIL were identified (30 Stage I, 35 Stage II). Three patients recurred in the inguinal region, (4.6%) and 11 patients (16.9%) recurred on the vulva. Two of the 11 patients died of disease, six patients are alive without evidence of disease after additional therapy. Five-year disease-free survival and overall survival were 66% and 97%, respectively. Risk of recurrence was not associated with smoking status, stage, or margin status. CONCLUSIONS: SupIL and vulvectomy for Stage I/II vulvar cancer have a low recurrence rate in the inguinal region when nodes are negative. The local recurrence rate (17%) is acceptable, and overall survival is good using this conservative approach.     

Cyclic AMP in normal and sympathetically aneural chick hearts during development

Basal levels of cyclic adenosine monophosphate (cAMP) were measured in embryonic chick hearts at various times during development. Basal cAMP was highest on incubation day 5 and decreased throughout the remaining incubation period. Cyclic AMP could not be stimulated above basal level by intravenous in ovo administration of isoproterenol or tyramine on incubation day 5; however, there was a decrease in cAMP 2 mins after intravenous injection which was identical to the decrease in cAMP in controls injected with saline. Heart rate decreased following intravenous injection of saline on incubation day 5, but a similar decrease was not observed following intravenous isoproterenol injection. Functional sympathetic innervation of the heart does not occur until incubation day 16, and this fact is responsible for the insensitivity to tyramine stimulation on incubation day 5. Although the level of cAMP could not be stimulated above basal level on incubation day 5, beta-antagonists caused a decrease in the level of cAMP, with no decrease in heart rate. These observations indicate that the beta-receptor is coupled to adenylate cyclase on incubation day 5 but it is questionable whether adenylate cyclase is effectively coupled to heart rate. Isoproterenol and tyramine caused a significant elevation in cardiac cAMP and heart rate on incubation day 17 following intravenous in ovo injection. Hearts made sympathetically aneural by removal of premigratory neural crest responded to isoproterenol but not tyramine on incubation day 17 which demonstrates that the morphologically aneural hearts are also functionally aneural.(ABSTRACT TRUNCATED AT 250 WORDS)