[Conventional intubation and laryngeal tube in cervical spine instability : Changes in the width of the dural sac in unfixed human body donors]. / Konventionelle Intubation und Larynxtubus bei Halswirbelsäuleninstabilität : Änderungen der Duralsackweite am unfixierten humanen Körperspender.

BACKGROUND: Airway management in patients with an unstable cervical spine requires a cautious approach if secondary damage is to be prevented but the question regarding the optimum method remains unresolved. The primary aim of the study was to investigate whether there were differences between intubation by conventional Macintosh laryngoscopy and placement of a laryngeal tube (LTS-D) with respect to dural sac compression on an unfixed human cadaver model with unstable injuries of the upper cervical spine. Secondary parameters that could be relevant in patients with unstable spinal injuries were also investigated. MATERIAL AND METHODS: Orotracheal intubation by conventional direct laryngoscopy using a Macintosh blade and placement of a laryngeal tube (LTS-D) were performed in six fresh human cadavers. The dural sac was filled with contrast dye to allow continuous myelography by lateral fluoroscopy. Changes in the width of the dural sac at the cervical segments (C) C0/C1 and the C1/C2 levels as well as secondary parameters (angulation, distraction, intervention time) were assessed in the intact spine as well as in the presence of combined atlanto-occipital dislocation and atlanto-axial instability. The intubation methods were considered independent and examined using the Mann-Whitney U­test. RESULTS: At the C0/C1 level in the intact spine, conventional laryngoscopy caused less reduction of the width of the dural sac than placement of the LTS-D (0.33 mm vs. 0.46 mm, p = 0.035); however, in the presence of combined atlanto-occipital dislocation and atlanto-axial instability, placement of the LTS-D caused less reduction in the width of the dural sac than conventional intubation (1.18 mm vs. 0.68 mm, p = 0.005). At the C1/C2 level no differences were found with respect to changes in the width of the dural sac, neither in the intact spine nor in combined atlanto-occipital dislocation and atlanto-axial instability. Conventional intubation caused more angulation than placement of the LTS-D at both levels measured. Both methods did not cause distraction. The intervention times for placement of the laryngeal tube were shorter. CONCLUSION: In an unfixed human cadaver model with combined atlanto-occipital dislocation and atlanto-axial instability, placement of the LTS-D caused less reduction in the width of the dural sac than conventional intubation at the level of the craniocervical junction. The LTS-D also caused less angulation and could be placed faster. It could therefore also be advantageous over conventional intubation in living patients with an unstable cervical spine.

Minimal prophylactic concentration of dietary zinc compounds in a mouse model of swine dysentery.

Dietary supplementation with 6000 mg of Zn2+/kg of feed has been shown to modify the clinicopathologic expression of Brachyspira hyodysenteriae infection in a laboratory mouse model of swine dysentery. However, this concentration impaired the body weight gain of the mice. The purpose of the present study was to determine a minimal prophylactic concentration of feed-grade zinc compounds that would not affect the growth of mice challenge-exposed with B. hyodysenteriae. A total of 440, 6- to 8-week-old, C3H/HeN mice were allocated randomly to groups and fed either a defined diet or a defined diet containing either 1000, 2000 or 4000 mg/kg ZnO, ZnSO4 or zinc-methionine for 7 days before intragastric inoculation with B. hyodysenteriae. From days 7 to 35 after inoculation, mice in each group were necropsied at weekly intervals for determination of body weight, presence of B. hyodysenteriae in the cecum, and histological assessment of cecal lesions. Only ZnO fed at 2000 mg/kg had a prophylactic effect against B. hyodysenteriae infection without affecting the body weight gain of the mice. The prophylactic effect of Zn2+ against infection with B. hyodysenteriae was also affected by the relative concentration of Fe2+ and Zn2+/Fe2+ ratio of the diet.

Precursor B-cell lymphoblastic lymphoma. A study of nine cases lacking blood and bone marrow involvement and review of the literature.

We describe 9 cases of precursor B-cell lymphoblastic lymphoma (LYL) without evidence of marrow or blood involvement. Four patients had superficial nodal disease, 2 cutaneous involvement, and 1 each ovarian, retroperitoneal, or tonsillar primary tumor. Six patients had limited disease; 3 patients were stage III. Immunophenotyping revealed a terminal deoxynucleotidyl transferase (TdT)-positive, immature B-cell population with variable expression of CD10, CD20, and CD45. All patients are in complete clinical remission (median follow-up, 14 months). A literature review yielded 105 patients with a diagnosis of precursor B-cell LYL based on less than 25% marrow involvement. Of these, 64% were younger than 18 years. Skin, lymph nodes, and bone were the most common sites of disease. Mediastinal involvement was uncommon. TdT, CD19, CD79a, CD10, and HLA-DR were the most frequently expressed antigens, while CD45 and CD20 were expressed in only two thirds of the cases. Cytogenetic analysis showed additional 21q material as a recurring karyotypic abnormality. At a median follow-up of 26 months, 74% of patients were alive; the median survival was 19 months for patients dying of disease. Comparison with precursor B-cell acute lymphoblastic leukemia showed several overlapping features, although distinct differences were identified.

Secondary myelodysplasia with monosomy 7 arising after treatment for acute lymphoblastic leukemia in childhood.

Monosomy 7 is recognized as a characteristic, clonal abnormality associated with acquired myelodysplasia (MDS) or acute myeloid leukemia (AML). It can occur as a late complication of cytotoxic therapy and is usually associated with exposure to alkylating agents or radiation therapy. We report two patients with therapy-related myelodysplasia (t-MDS) associated with monosomy 7 occurring in children after completion of therapy for acute lymphoblastic leukemia (ALL). Both children were noted to have t-MDS with monosomy 7 at the time of cessation of chemotherapy. Neither child had received an alkylating agent or radiation therapy during treatment. One child had a unique dicentric marker chromosome that was shown by fluorescent in situ hybridization to be derived from chromosome 7. This report emphasizes the need to identify and minimize therapy-related side effects without compromising cure rates.

New recurring cytogenetic abnormalities and association of blast cell karyotypes with prognosis in childhood T-cell acute lymphoblastic leukemia: a pediatric oncology group report of 343 cases.

To further define the cytogenetic differences between B-cell lineage (B-lineage) acute lymphoblastic leukemia (ALL) and T-cell lineage ALL (T-ALL) and to determine the prognostic value of cytogenetics in childhood T-ALL, the blast cell karyotypes of 343 cases of pediatric T-ALL, the largest series reported to date, were evaluated. Cytogenetics were performed in a single central laboratory, and the children were treated using a single Pediatric Oncology Group protocol. Clear differences between the karyotypic characteristics of B-lineage ALL and T-ALL were confirmed. This study suggests that there may be survival differences associated with some T-ALL blast cell karyotypes. Better survival is associated with only normal karyotypes and with t(10;14) (translocation of chromosomes 10 and 14); worse survival is associated with the presence of any derivative chromosome. Two new recurring chromosome aberrations previously not reported in T-ALL were found: del(1)(p22) and t(8;12)(q13;p13). Ten aberrations found in this series, which were reported only once previously in T-ALL, can now be considered recurring abnormalities in T-ALL. All 12 of these new recurring aberrations are targets for discovery and characterization of new genes that are important in T-cell development and leukemogenesis.

Gas chromatography/mass spectrometry identification and quantification of isazophos in a famphur pour-on and in bovine tissues after a toxic exposure.

A sample identified as "Warbex pour-on," expected to contain 13.2% famphur, and bovine tissue samples from 2 heifers that died after exhibiting signs of organophosphate intoxication were analyzed by gas chromatography/mass spectrometry (GC/MS). A product formulation problem was suspected because brain cholinesterase activities were depressed in both animals. Electron impact (EI) GC/MS of the pour-on revealed 9.7% famphur and an unidentified peak with approximately 76% of the peak area of the famphur. The unidentified peak showed a molecular ion at m/z 313, with a single Cl isotope cluster. Methane chemical ionization (MeCI) MS confirmed the molecular weight at 313 (1 Cl). A search on the molecular formula C9H17N3O3PSCl yielded a single match, isazophos. EI and MeCI GC/MS of reference isazophos confirmed the identity of the suspect peak. The concentration of isazophos in the pour-on was determined to be 6.0%. Famphur and isazophos were identified by their EI spectra and GC retention times in extracts of liver and brain from the 2 deceased animals. A GC/MS procedure utilizing selected ion monitoring (SIM) was developed for quantification of isazophos in liver, kidney, muscle, and fat of additional affected animals sacrificed at various times after exposure. Isazophos remained in animal tissues for as long as 94 days after topical exposure. Isazophos was present in fetal liver 70 days after exposure of the dam. High levels (6-3,500 ppm) of isazophos and famphur remained on the skin at 39 days postexposure.

Extensive analysis of mosaicism in a case of Turner syndrome: the experience of 287 cytogenetic laboratories. College of American Pathologists/American College of Medical Genetics Cytogenetics Resource Committee.

OBJECTIVE: To assemble and interpret karyotype data provided as part of the College of American Pathologists/American College of Medical Genetics Cytogenetics Proficiency Testing Program. DATA SOURCES, EXTRACTION, AND SYNTHESIS: The Cytogenetics Resource Committee requested data on all cells analyzed in a 1994 whole-blood specimen challenge. In that study, 287 participating laboratories analyzed a total of 14297 cells derived from a sample drawn from an adult donor with Turner syndrome. This individual had previously been found to have mosaicism, including cell lines with X structural anomalies along with monosomy X, making this an excellent challenge for a multicenter cytogenetic survey. RESULTS AND CONCLUSIONS: Analysis of the data from this extensive study revealed mosaicism of up to 10 different sex chromosome complements involving the X chromosome with and without a small ring X or a derivative X chromosome. In the routine cytogenetic analysis performed by the participating laboratories, cell lines observed, in decreasing order of prevalence, included 45,X (n = 8357 cells), 46,X,r(X) (n = 3597), 46,X,der(X)t(X;X) (n = 2237), 46,XX (n = 93), 47,X,r(X),r(X) (n = 5), 47,X,der (X)t(X;X),der(X)t(X;X) (n = 3), 47,XX,r(X) (n = 2), and one observation each of 47,XX,der(X)t(X;X), 47,X,der(X)t (X;X),r(X), and 47,XXX. Our molecular cytogenetic data, as well as detailed analysis of G-banded chromosomes, suggest the nomenclature for these 2 abnormal X chromosomes as r(X)(p11.3q21.3) and der(X)t(X;X)(p11.3;q21.3), and we discuss models for the concomitant formation of these 2 entities. Both the degree of analysis and the extensive mosaicism that was discovered in this study are exceptional, and similar reported cases as well as possible mechanisms for the observed X chromosome instability are reviewed.

Effects of supplementation of organic and inorganic combinations of copper, cobalt, manganese, and zinc above nutrient requirement levels on postpartum two-year-old cows.

The objective of this study was to determine whether a combination of Cu, Co, Mn, and Zn in an organic or inorganic form fed at higher than nutrient recommendations for 2-yr-old cows from calving to breeding would affect pregnancy rate, calving date, calf performance, and cow liver and serum mineral concentrations. Crossbred 2-yr-old cows were used after calving in 1994 (n = 127) and 1995 (n = 109). Cows were blocked by calving date to one of three treatments: 1) no supplemental minerals (CTL), 2) organic minerals (ORG), or 3) inorganic minerals (ING). Minerals were fed for the same daily intake for both organic and inorganic treatments: Cu (125 mg), Co (25 mg), Mn (200 mg), and Zn (360 mg). Cows were individually fed a mineral-protein supplement with grass hay from calving (February-March) to before breeding (May 15). Hay intakes were calculated using chromium oxide boluses to determine fecal output. Fecal excretion of minerals was calculated following trace element analysis of feces. Liver biopsies were obtained before calving, after calving (start of supplementation), at the end of supplementation, and in midsummer. Over 2 yr, more cows did not become pregnant (P < .01) in ORG (11/78) and ING (11/78) treatments than in CTL (0/80) treatments. A treatment x year interaction was found for day of conception. Cows in the ORG group conceived later (P < .01) than cows in the ING or CTL groups in 1994. In 1995, there was no difference (P > .10) in day of conception among groups. Liver Zn and Mn concentrations were not different (P > .10) and Cu concentrations increased (P < .01) for the ORG and ING groups. Cows in the ORG and ING groups had higher (P < .01) concentrations of Cu, Mn, and Zn in the feces than the CTL cows. Trace elements in the feces did not differ for ORG and ING groups. Results indicate that combinations of Cu, Co, Mn, and Zn fed at higher levels than are required reduced reproductive performance.

Case report of a 22-week fetus with 47,XXX karyotype and multiple lower mesodermal defects.

A 22-week stillborn fetus with 47,XXX karyotype had lower mesodermal defects consisting of irregular fusion of the sacral vertebrae, anal agenesis, multicystic dysplasia of a horseshoe kidney, a single umbilical artery, dysplastic ovaries, and uterine hypoplasia. This case provides additional evidence for an association between trisomy X and genitourinary defects including lower mesodermal defects sequence.

Characterization of a breast cancer cell line derived from a germ-line BRCA1 mutation carrier.

A tumor cell line, HCC1937, was established from a primary breast carcinoma from a 24-year-old patient with a germ-line BRCA1 mutation. A corresponding B-lymphoblastoid cell line was established from the patient's peripheral blood lymphocytes. BRCA1 analysis revealed that the tumor cell line is homozygous for the BRCA1 5382insC mutation, whereas the patient's lymphocyte DNA is heterozygous for the same mutation, as are at least two other family members' lymphocyte DNA. The tumor cell line is marked by multiple additional genetic changes including a high degree of aneuploidy, an acquired mutation of TP53 with wild-type allele loss, an acquired homozygous deletion of the PTEN gene, and loss of heterozygosity at multiple loci known to be involved in the pathogenesis of breast cancer. Comparison of the primary tumor with the cell line revealed the same BRCA1 mutation and an identical pattern of allele loss at multiple loci, indicating that the cell line had maintained many of the properties of the original tumor. This breast tumor-derived cell line may provide a useful model system for the study of familial breast cancer pathogenesis and for elucidating BRCA1 function and localization.

The first recurring chromosome translocation in hepatoblastoma: der(4)t(1;4)(q12;q34).

We report four cases of hepatoblastoma with a derivative chromosome 4 from an unbalanced translocation between the long arms of chromosomes 1 and 4, an aberration reported only rarely in isolated cases of other types of neoplasms. The abnormality in three hepatoblastomas was der(4)t(1;4)(q12;q34), whereas the fourth case appeared to have a der(4)t(q25;q32). All had hyperdiploid tumor karyotypes; however, in the case with t(q25;q32), the der(4) was the only abnormality in the stemline. We speculate that the oncogenetic event in our cases may be the loss of a gene on distal 4q or their alteration by juxtaposition to 1q12 heterochromatin.

All patients with the T(11;16)(q23;p13.3) that involves MLL and CBP have treatment-related hematologic disorders.

The involvement of 11q23-balanced translocations in acute leukemia after treatment with drugs that inhibit the function of DNA topoisomerase II (topo II) is being recognized with increasing frequency. We and others have shown that the gene at 11q23 that is involved in all of these treatment-related leukemias is MLL (also called ALL1, Htrx, and HRX). In general, the translocations in these leukemias are the same as those occurring in de novo leukemia [eg, t(9;11), t(11;19), and t(4;11)], with the treatment-related leukemias accounting for no more than 5% to 10% of any particular translocation type. We have cloned the t(11;16)(q23;p13.3) and have shown that it involves MLL and CBP (CREB binding protein). The CBP gene was recently identified as a partner gene in the t(8;16) that occurs in acute myelomonocytic leukemia (AML-M4) de novo and rarely in treatment-related acute myeloid leukemia. We have studied eight t(11;16) patients, all of whom had prior therapy with drugs targetting topo II with fluorescence in situ hybridization (FISH) using a probe for MLL and a cosmid contig covering the CBP gene. Both probes were split in all eight patients and the two derivative (der) chromosomes were each labeled with both probes. Use of an approximately 100-kb PAC located at the breakpoint of chromosome 16 from one patient revealed some variability in the breakpoint because it was on the der(16) in three patients, on the der(11) in another, and split in four others. We assume that the critical fusion gene is 5'MLL/3'CBP. Our series of patients is unusual because three of them presented with a myelodysplastic syndrome (MDS) most similar to chronic myelomonocytic leukemia (CMMoL) and one other had dyserythropoiesis; MDS is rarely seen in 11q23 translocations either de novo or with t-AML. Using FISH and these same probes to analyze the lineage of bone marrow cells from one patient with CMMoL, we showed that all the mature monocytes contained the fusion genes as did some of the granulocytes and erythroblasts; none of the lymphocytes contained the fusion gene. The function of MLL is not well understood, but many domains could target the MLL protein to particular chromatin complexes. CBP is an adapter protein that is involved in regulating transcription. It is also involved in histone acetylation, which is thought to contribute to an increased level of gene expression. The fusion gene could alter the CBP protein such that it is constitutively active; alternatively, it could modify the chromatin-association functions of MLL.

Cytogenetics as an adjunct in establishing a definitive diagnosis of synovial sarcoma by fine-needle aspiration.

BACKGROUND: Synovial sarcomas account for up to 10% of all soft tissue sarcomas and are characterized by a specific chromosomal abnormality, t(X; 18)(p11.2; q11.2), that is observed in both monophasic and biphasic variants. METHODS: A 9-cm suprascapular mass in a 43-year-old man and a 10-cm leg mass in a 45-year-old man were aspirated. Rapid evaluation of air-dried Diff-Quick-stained smears in both cases showed a malignant spindle cell lesion. Cytogenetic material was collected in RPMI 1640 tissue culture medium. Additional alcohol-fixed slides and cell blocks were prepared on both cases. Immunohistochemical studies were performed on paraffin embedded cell block sections in both cases whereas electron microscopic examination was performed in one case. G-banded chromosome preparations from tumor cells were made after short term culture using standard cytogenetic techniques. RESULTS: Evaluation of the smears showed densely cellular and tightly cohesive malignant spindle cells without discernible epithelial differentiation. A differential diagnosis of synovial sarcoma, leiomyosarcoma, malignant schwannoma, and fibrosarcoma was considered. Immunohistochemical stains and electron microscopy were noncharacteristic and noncontributory in establishing a diagnosis. However, cytogenetic results revealed a t(X; 18)(p11.2; q11.2) translocation, thus establishing the diagnosis of synovial sarcoma. Both patients were offered definitive therapy based on fine-needle aspiration (FNA) diagnosis only. CONCLUSIONS: FNA can be used effectively to obtain karyotypes of sarcomas. The specific cytogenetic abnormality associated with synovial sarcoma provides an opportunity to make definitive diagnoses using a combination of FNA and cytogenetics, obviating open surgical biopsy preceding therapy.

Prune-belly syndrome and other anomalies in a stillborn fetus with a ring X chromosome lacking XIST.

Ring X chromosomes that lack the X inactivation center and fail to be inactivated have been implicated as a cause of mental retardation and multiple congenital anomalies. We report on a stillborn fetus with karyotype mos45,X/46,X,r(X) and early urethral obstruction or prune-belly sequence, single umbilical artery, limb deficiency, horseshoe kidney, cardiac hypertrophy, persistent left superior vena cava, and axial skeleton abnormalities. Fluorescent in situ hydridization (FISH) studies confirmed that the ring chromosome is X-derived and demonstrated that it lacks the XIST locus. The findings in this fetus are discussed with regard to the spectrum of phenotypes associated with monosomy X and small ring X chromosomes.

Pilot studies for proficiency testing using fluorescence in situ hybridization with chromosome-specific DNA probes: a College of American Pathologists/American College of Medical Genetics Program.

Fluorescence in situ hybridization using chromosome-specific DNA probes is rapidly becoming part of clinical laboratory practice for certain congenital and neoplastic disorders. Current legislation requires proficiency testing for clinical laboratory studies. To evaluate the efficacy of fluorescence in situ hybridization proficiency testing, we invited 19 representative institutions to participate in three pilot studies. One study used probes for the X and Y chromosomes to evaluate metaphase spreads and interphase nuclei. Another study used probes for bcr and abl to detect bcr/abl fusion in interphase nuclei in chronic myelogenous leukemia. The third study used a D22S75 probe to detect microdeletions in metaphase spreads from a patient with velocardiofacial syndrome. The results of these studies demonstrate that proficiency testing with fluorescence in situ hybridization is attainable using either metaphase or interphase preparations, and that either microscope slides or fixed cell pellets are suitable.

Report of a complex karyotype in recurrent metastatic fibrolamellar hepatocellular carcinoma and a review of hepatocellular carcinoma cytogenetics.

Metastatic fibrolamellar hepatocellular carcinoma (HCC) was detected in the abdominal lymph nodes of an adolescent male after resection of the primary tumor. No dividing cells were isolated from attempted cytogenetic studies of the primary tumor. However, cytogenetic analysis of lymph node metastases detected 9 and 12 months after partial hepatectomy revealed abnormal hypertriploid karyotypes, with a suggestion of clonal evolution: 62-92 < 3n >,XX, -Y, +3, +6, +6, +7, +7, +8, +10, +13, +15, +16, +20, -21, -22, +mar1 x 2, +mar[cp6]/46,XY[8] and 78 < 3n >,XX, -Y,der(1)t(1;1)(p36.1;q21), +4, +6, +6, +7, +7,i(8)(q10), +10, +15, +20, -21, -22, +mar1 x 2, +mar2[3]/46, XY[17], respectively. Karyotypes of this variant of HCC have not been reported previously. The cytogenetics of HCC are reviewed.

Primary body cavity-based AIDS-related lymphomas.

Five patients with advanced AIDS developed a unique type of high grade primary body cavity-based non-Hodgkin's lymphoma (NHL). The lymphomas were exclusively in serous effusions with no detectable mass disease in the body cavities and no lymphadenopathy or organomegaly. All of the lymphomas exhibited virtually identical morphology, which could not be precisely classified, but appeared to bridge features of large cell immunoblastic and anaplastic large cell lymphomas. Immunophenotypically the lymphoma cells lacked expression of any B- or T-lymphocyte antigens, but expressed CD45 and the activation antigens CD30, CD38, CD71, and HLA-DR. Clonally rearranged immunoglobulin heavy chain and kappa light chain genes were identified by Southern blot analysis. Molecular studies also revealed Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) genomes and germline configuration of the c-myc protooncogene. In two cases studied cytogenetically, the lymphoma cells manifested complex chromosome abnormalities. These lymphomas are clinically and biologically unique and found predominantly in patients with advanced AIDS, in many cases with pre-existing Kaposi's sarcoma.

Identification and molecular confirmation of a small chromosome 10q duplication [dir dup(10)(q24.2-->q24.3)] inherited from a mother mosaic for the abnormality.

We describe a family in which two siblings exhibited developmental delay, reduced muscle tone and mild muscle weakness. Cytogenetic evaluation demonstrated that both children had a tandem duplication of a small portion of the long arm of chromosome 10 [46,XX or XY,dir dup(10)(q24.2-->q24.3)], inherited from their clinically normal mother, who was found to be mosaic for the duplicated chromosome 10. Fluorescence in situ hybridization approaches, including total chromosome painting and the use of regional specific cosmid probes, were used to confirm the chromosome 10q origin of the duplicated material. This is the smallest confirmed duplication of this portion of chromosome 10 reported to date.