Reassessment of the utility of fetal umbilical vein diameter in the management of isoimmunization

High-resolution ultrasound has been recently introduced in the management of many conditions in which the fetus is at risk. In the detection of severe erythroblastosis fetalis, sonographic evaluation of fetal anatomic changes is being used in association with amniotic fluid spectral analysis to assess the degree of the hemolytic process. In 1981 it was reported that sonographically detected umbilical vein dilatation, resulting from hepatic congestion, could be used as a sign of impending fetal compromise. We analyzed data obtained from 47 patients in a total of 76 examinations, including sonographic measurement of umbilical vein diameter, associated ultrasound findings, results of amniotic fluid spectral analyses, and neonatal outcome. The collected data, divided in two groups according to the results of amniotic fluid spectral analyses (less than high zone II and greater than or equal to high zone II), showed that the sonographic measurement of umbilical vein diameter does not differ significantly between the two groups (p greater than 0.05). Therefore, the present and relatively large series demonstrates that dimensions of the umbilical vein cannot be used as a reliable predictor of worsening fetal disease.     

Experimental reproduction of the runting-stunting syndrome of broiler chickens

A model for the reproduction of the runting-stunting syndrome (RSS) of broiler chickens is described. In this model, groups of at least 90 day-old broiler chickens were inoculated (per os) with various tissue homo-genates or virus preparations. During the first week post-inoculation, birds were examined for the development of histopathological changes in their intestines. At day 14 post-inoculation, the remaining birds were weighed and tested for elevations in plasma amylase activity and examined for the development of pancreatic atrophy. Bacteria-free intestinal and pancreatic homogenates from chickens of different ages, taken from flocks which developed RSS, regularly induced a lower mean live-weight in treated birds. Of these, only intestinal homogenates prepared from 5-day-old birds induced intestinal lesions, lowered mean live-weight and increased the incidence of both elevated plasma amylase activity and pancreatic atrophy. These changes were more marked in birds exposed to short periods of sub-optimal temperatures during the first week post-inoculation. An ultracentri-fuged pellet prepared from this intestinal homogehate, was also found to induce an increased incidence of pancreatic atrophy in treated birds. These studies suggest that the causative agent(s) of RSS is an as yet unidentified virus, and that the effects of this infection are greater in birds subjected to stress, such as sub-optimal temperature exposure, within the first week of hatch.     

Molecular cytogenetic abnormalities in patients with concurrent chronic lymphocytic leukemia and multiple myeloma shown by interphase fluorescence in situ hybridization: evidence of distinct clonal origin

The coexistence of chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) is rare, and there is no consensus about the clonal relationship of the two disorders when they occur in the same individual. We investigated chromosomal abnormalities in two patients with concurrent CLL and MM using interphase fluorescence in situ hybridization (FISH) with a panel of region-specific DNA probes. In patient 1, the clonal plasma cells harbored IgH translocations (14q32); however, FISH with probes for the four most frequent IgH partner genes in MM (CCND1, FGFR3, MAF, and MYC) did not detect translocations involving any of them. The CLL cells were characterized by deletions of 13q14, 11q23, and 17p13, as well as trisomy 12, none of which were found in the MM cells. In patient 2, deletions of 13q14 and 17p13 were detected in CLL cells, but no cytogenetic abnormalities were found in the MM cells. Both patients had relapsed MM following chemotherapy and had autologous stem-cell transplant, whereas their CLL has been stable and not requiring treatment. Our results show that the cytogenetic profiles differ between CLL and MM within the same patients, and provide evidence for two distinct malignant clones in both patients.     

Use of a T cell-specific monoclonal antibody, T10B9, in a novel allogeneic stem cell transplantation protocol for hematologic malignancy high-risk patients.

To reduce the toxicity of traditional conditioning regimens for allogeneic stem cell transplantation (allo-SCT), we used single-agent chemotherapy conditioning with either busulfan (total cumulative dose, 16 mg/kg) or melphalan (200 to 240 mg/m 2 ), followed by the anti-T cell-specific monoclonal antibody T10B9 (MEDI-500) daily for 3 days. T cell-replete SCT was performed from HLA-identical sibling donors. Acute graft-versus-host disease (aGVHD) prophylaxis consisted of 7 additional days of T10B9 and delayed onset of cyclosporine (ie, on day +4 or +5). Twenty-six high-risk hematologic malignancy patients were entered onto this study. All 24 patients who survived longer than 8 days engrafted, although 1 patient experienced late graft failure. Deaths occurred in 21 of 26 patients because of infection (n = 7), progression/recurrence of primary disease (n = 6), aGVHD (n = 4), regimen-related toxicity (n = 1), and other causes (n = 3). Five of these patients are enjoying disease-free survival with a median survival of 1193 days after allo-SCT. The conditioning regimen induced modulation of surface expression of CD3 (but not CD4 or CD8) and was associated with decreasing tumor necrosis factor-alpha (but not interleukin-6) serum levels. In conclusion, single-agent chemotherapy conditioning with T10B9 produced durable engraftment and long-term survival in some patients who would not have qualified for a traditional allo-SCT.     

IL‐27p28 is essential for parent‐to‐F1 acute graft‐versus‐host disease

Acute graft versus host disease (aGVHD) remains a life‐threatening complication of bone marrow transplantation. Here we show that IL‐27, a member of the IL‐12 cytokine family, plays an essential role in a parent‐to‐F1 murine aGVHD model, using B6 mice as parents and B6D2 mice as F1 recipients. IL‐27 is transiently detectable in the serum of B6D2 recipients of B6 spleen cells, with a peak at day 10. Treatment with anti‐IL‐27p28 mAb MM27.7B1 (αp28Ab), at the time of and six days after B6 cell transfer, blocked GVHD. Protection was associated with host cell survival and undiminished engraftment of donor cells, lack of host B‐cell depletion, increased Th2‐type immunoglobulin production, a decrease in serum IFN‐γ, a drop in anti‐H‐2D^d cytotoxic T lymphocyte activity and an increase in Foxp3⁺ T cells. We therefore conclude that IL‐27 plays a critical role in the parent‐to‐F1 model of aGVHD and that blocking IL‐27 could have therapeutic relevance.     

Detection of chromosomes and estimation of aneuploidy in human spermatozoa using fluorescence in-situ hybridization

The development and application of fluorescence in-situ hybridization (FISH) has opened the way for comprehensive studies on numerical chromosome abnormalities in human spermatozoa. FISH can be rapidly applied to large numbers of spermatozoa and thus overcomes the major limitation of karyotyping spermatozoa after penetration of zona-free hamster oocytes. The simultaneous hybridization of two or more chromosome-specific probes to spermatozoa and subsequent detection of the bound probes using different fluorescent detection systems enables two or more chromosomes to be localized simultaneously in the same spermatozoon and provides a technique for undertaking reasonable estimates of aneuploidy. The most commonly used probes are those which bind to the centromeric region of specific chromosomes. Most studies to date have concentrated on estimating aneuploidy in spermatozoa from normospermic men, although reports are beginning to appear on aneuploidy in spermatozoa from subfertile and infertile men. Multi- probe FISH studies have generally reported disomy (hyperhaploidy) estimates of 0.05-0.2% per chromosome. There is preliminary evidence that some chromosomes such as X, Y and 21 are predisposed towards higher rates of non-disjunction during spermatogenesis. There are also suggestions of inter-donor variability in aneuploidy frequencies for specific chromosomes, although this requires confirmation in larger studies. While FISH is clearly a powerful technique that has many applications in reproductive medicine, it must also be realized that it does have limitations and the technology itself is still evolving and has yet to be fully validated on spermatozoa.      

Identification of MEN1 gene mutations in sporadic carcinoid tumors of the lung

Lung carcinoids occur sporadically and rarely in association with multiple endocrine neoplasia type 1 (MEN1). There are no well defined genetic abnormalities known to occur in these tumors. We studied 11 sporadic lung carcinoids for loss of heterozygosity (LOH) at the locus of the MEN1 gene on chromosome 11q13, and for mutations of the MEN1 gene using dideoxy fingerprinting. Additionally, a lung carcinoid from a MEN1 patient was studied. In four of 11 (36%) sporadic tumors, both copies of the MEN1 gene were inactivated. All four tumors showed the presence of a MEN1 gene mutation and loss of the other allele. Observed mutations included a 1 bp insertion, a 1 bp deletion, a 13 bp deletion and a single nucleotide substitution affecting a donor splice site. Each mutation predicts truncation or potentially complete loss of menin. The remaining seven tumors showed neither the presence of a MEN1 gene mutation nor 11q13 LOH. The tumor from the MEN1 patient showed LOH at chromosome 11q13 and a complex germline MEN1 gene mutation. The data implicate the MEN1 gene in the pathogenesis of sporadic lung carcinoids, representing the first defined genetic alteration in these tumors.      

De novo duplication of MECP2 in a girl with mental retardation and no obvious dysmorphic features

Makrythanasis P, Moix I, Gimelli S, Fluss J, Aliferis K, Antonarakis SE, Morris MA, Béna F, Bottani A. De novo duplication of MECP2 in a girl with mental retardation and no obvious dysmorphic features. Loss‐of‐function mutations of MECP2 are responsible for Rett syndrome (RTT), an X‐linked neurodevelopmental disorder affecting mainly girls. The availability of MECP2 testing has led to the identification of such mutations in girls with atypical RTT features and the recognition of milder forms. Furthermore, duplication of the entire gene has recently been described in boys with mental retardation and recurrent infections. We describe a girl with a heterozygous de novo MECP2 duplication. The patient, at the age of 19, has mental retardation with no autistic features. She is friendly but gets frequently anxious. She has neither dysmorphic features nor malformations. Her motor development was delayed with walking at 20 months. Speech is fluid with good pronunciation but is simple and repetitive. Diagnosis was made after single‐strand conformation analysis (SSCA) and multiplex ligation‐dependent probe amplification (MLPA) analysis of MECP2. Array comparative genomic hybridization (aCGH) analysis showed a duplication of 29 kb including MECP2 and part of IRAK1. Fluorescent in situ hybridization (FISH) has revealed that the duplicated region is inserted near the telomere of the short arm of chromosome 10. X‐chromosome inactivation in leukocyte DNA was not skewed. We conclude that it is likely that this MECP2 duplication is responsible for the mental retardation in this patient. This case broadens the phenotypic spectrum of MECP2 abnormalities with consequent implication in diagnosis and genetic counselling of girls with non‐syndromic mental retardation.