Fetal t(5p;21q) misdiagnosed as monosomy 21: a plea for in situ hybridization studies.

We report a case of 45,XY,-5,-21,+der (5) t(5;21) (p13 or p14;q11.2 or q21) that was prenatally misdiagnosed as complete monosomy 21 and terminated at 24 weeks of gestation. Subsequent fluorescence in situ hybridization studies with a chromosome 21 painting probe documented the cryptic unbalanced translocation.

Duplication (6q) syndrome diagnosed in utero.

We report on a case of partial duplication 6q detected ultrasonographically. The clinical picture noted in utero is consistent with the adult phenotype previously reported in the literature.

Inconsistencies in pedigree symbols in human genetics publications: a need for standardization.

To determine consistency in usage of pedigree symbols by genetics professionals, we reviewed pedigrees printed in 10 human genetic and medical journals and 24 medical genetics textbooks. We found no consistent symbolization for common situations such as pregnancy, spontaneous abortion, death, or test results. Inconsistency in pedigree design can create difficulties in the interpretation of family studies and detract from the pedigree's basic strength of simple and accurate communication of medical information. We recommend the development of standard pedigree symbols, and their incorporation into genetic publications, professional genetics training programs, pedigree software programs, and genetic board examinations.

Enhancement of chemotactic migration by the local anesthetic tetracaine.

The local amine anesthetic tetracaine added to a suspension of guinea pig or human neutrophilic granulocytes inhibited their random migration in Boyden chambers, but increased their chemotactic migration towards the chemotactic tripeptide f-Met-Leu-Phe, complement-activated normal guinea pig serum, and the eosinophil chemotactic factor ECF. Tetracaine not only increased the distance migrated by the leading cells, it also caused more cells to leave the upper filter surface and to migrate into the filter. The effect required the presence of the drug; cells preincubated with tetracaine and washed did not differ from control cells. It is suggested that tetracaine specifically enhanced a mechanism operative in a cell's response to a concentration gradient of a chemotactic factor.

Generation of chemotactic activity in normal guinea pig serum by treatment with acid.

Chemotactic activity was generated in normal guinea pig serum following titration of the pH to 4 by the addition of hydrochloric acid. The process was time and temperature dependent. The chemotactic material was stable when kept at pH 4, but decayed within one or two days when the pH was raised to neutrality. Fractionation of the acid-treated serum by Sephacryl S-200 gel filtration revealed two peaks of activity, one eluting with molecules larger than bovine serum albumin, the other in the 13,000 MW range. After heating the serum at 56 degrees C for 60 min no chemotactic activity could be generated by acid, while EDTA, EGTA, hydrazine and the enzyme inhibitors benzamidine, EACA, PMSF, and pepstatin, added to the serum prior to the addition of acid, had no effect. The results suggest that chemotactic factors, remarkably similar to the products of complement activation, could be generated in guinea pig serum by a mechanism differing from the classical or the alternative complement activation pathways.

Adherence and migration of guinea pig granulocytes after enzyme treatment of the cell surface.

Glycogen-induced polymorphonuclear granulocytes (PMN) from the peritoneal cavity of guinea pigs were examined (1) for their adherence to nylon fibers in the absence and presence of the adherence-enhancing chemotactic peptide formyl-methionyl-leucyl-phenylalanine (f-MLP), (2) for their random migration through the filter of a Boyden chamber and (3) for their chemotactic migration towards f-MLP. The cells were analyzed before and after treatment with the enzymes neuraminidase, papain and trypsin. PMN adhesiveness was increased by neuraminidase digestion but reduced by treatment with the proteolytic enzymes. Neuraminidase and trypsin had no effect on cell migration, while papain reduced random migration without affecting f-MLP-induced chemotaxis. The data suggest that the type of adherence measured by the nylon fiber method differs from the temporary attachment of cells migrating through a chemotaxis filter towards an attracting substance.

The effects of bovine serum albumin and the chemotactic peptide formyl-methionyl-leucyl-phenylalanine on the adherence of guinea pig polymorphonuclear leukocytes to nylon fiber columns.

Glycogen-induced guinea pig polymorphonuclear leukocytes were passed over nylon fiber columns and adherent and nonadherent fractions were quantitated by determination of hexosaminidase activity after lysis with Triton. Bovine serum albumin reduced the number of adherent cells at 37 degrees C as well as 4 degrees C, after fixation of the cells with glutaraldehyde, and in the absence and presence of EDTA. The chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine increased the adherent cell fraction at 37 degrees C, but failed to do so at 4 degrees C, after glutaraldehyde fixation and in the presence of 10 mmol/EDTA. Since metabolic inhibitors had no effect on the rate of adherence it is suggested that increased cell deformability is responsible for chemotactic factor-induced increased cell attachment to nylon fibers.

Functional properties of a novel neutrophil chemotactic factor derived from human monocytes.

Neutrophilic granulocytes (PMN) are attracted to sites of inflammation by chemotactic factors, the most potent of which are the complement split product C5a, the leukotriene B4 and the bacterial chemotactic factor-related tripeptide formyl-methionyl-leucyl-phenylalanine (FMLP). In addition to inducing directed migration, these agents increase the adherence of PMN to synthetic surfaces and endothelial cells; some stimulate an oxidative burst and the production of reactive oxygen derivatives, and they may be involved in the release of granule constituents. Here, we describe studies on the activities stimulated by a novel monocyte-derived chemotaxin (MOC). Human MOC attracted human PMN, but not monocytes or eosinophils. Like all chemotactic agents, it increased the adherence of PMN on nylon fibers. In contrast to other chemotactic factors it did not stimulate the release of superoxide anion regardless whether the cells were in suspension or adherent on nylon fibers. There was no release of the primary granule enzyme glucosaminidase or the secondary granule component vitamin B12-binding protein in the absence or presence of cytochalasin B. The results suggest that MOC is a unique chemotactic agent with properties different from the most potent chemotactic factors C5a, LTB4 and FMLP. The delayed release from macrophages suggests its involvement in protracted and chronic inflammatory reactions.

The effect of a sulfonated shale oil extract (Ichthyol) on the migration of human neutrophilic granulocytes in vitro.

The sulfonated shale oil extract, Ichthyol, was studied for its effect on the migration of human neutrophilic granulocytes by the Boyden chamber technique. When presented to the cells in a concentration gradient, Ichthyol induced a directed migration. There was little or no chemokinetic effect of Ichthyol when added to the cell compartment of the Boyden chamber. The chemotactic migration towards the tripeptides, formyl-methionyl-leucyl-phenylalanine and formyl-norleucyl-leucyl-phenylalanine, towards the arachidonic acid-derived eosinophil chemotactic factor released from neutrophils by the ionophore A 23187, and towards complement-derived chemotactic activity of normal human serum was inhibited or abrogated by Ichthyol. The Ichthyol effect on f-MLP chemotaxis could be partly overcome by excessive f-MLP concentrations. It was reversible when Ichthyol-incubated cells were washed and resuspended in regular buffer. It is suggested that various substances contained in Ichthyol interacted with either the chemotactic factors or the cell membrane or both and thus blocked cell stimulation. The results could help to explain the cell accumulation and abscess formation observed with Ichthyol in inflammatory skin lesions and the anti-inflammatory properties of the drug.

Inhibitory effect of sulfonated shale oils (ammonium bituminosulfonate) on the stimulation of neutrophilic granulocytes by the chemotactic tripeptide f-Met-Leu-Phe.

Sulfonated shale oils (ammonium bituminosulfonate, ichthammol, Ichthyol), shown previously to induce the directed migration of human neutrophils in Boyden chambers and to inhibit the directed migration towards the chemotactic factors C5a, LTB4, and f-Met-Leu-Phe, were studied for their effect on other neutrophil functions, which are stimulated by chemotactic factors. Like other chemotactic factors ammonium bituminosulfonate increased the adherence of neutrophils to nylon fibers, but it did not induce the release of the primary granule enzyme glucosaminidase from cytochalasin B-treated cells and it did not stimulate the production of oxygen radicals as measured by lucigenin-dependent chemiluminescence if studied under nontoxic conditions. When added together with the chemotactic tripeptide f-Met-Leu-Phe, ammonium bituminosulfonate inhibited adherence augmentation, enzyme release, and oxygen-radical production induced by the chemotactic factor. The results indicate that ammonium bituminosulfonate not only inhibited chemotactic migration but the whole spectrum of neutrophil functions induced by a chemotactic factor.

Production of superoxide anion and hydrogen peroxide by human neutrophilic granulocytes during chemotactic migration towards f-Met-Leu-Phe, C5a, leukotriene B4, monocyte-derived chemotaxin/IL-8 and platelet-activating factor.

During the chemotactic migration of human neutrophilic granulocytes towards the chemotactic factors f-Met-Leu-Phe, C5a, leukotriene B4 (LTB4), monocyte-derived chemotaxin (MOC/IL-8) and platelet-activating factor (PAF) in Boyden chambers, the production of superoxide anion and hydrogen peroxide was measured by superoxide dismutase-inhibitable cytochrome C reduction and oxidation of p-OH-phenylacetic acid, respectively. With the exception of 10(-6) M PAF, none of the factors at optimal chemotactic concentrations induced the production of O2- or H2O2 in amounts significantly different from neutrophilic granulocytes migrating at random. At 20-50 times the optimal chemotactic concentration some O2- and H2O2 production was observed with f-Met-Leu-Phe, C5a and LTB4, but not with MOC/IL-8. Superoxide dismutase, catalase or a combination of the two added to both compartments of the Boyden chambers did not affect the random or chemotactic migration towards any of the chemotactic factors. The results suggest that chemotactic migration and the production of reactive oxygen metabolites by human neutrophilic granulocytes are unrelated events.

Differential effects of nylon fibre adherence on the production of superoxide anion by human polymorphonuclear neutrophilic granulocytes stimulated with chemoattractants, ionophore A23187 and phorbol myristate acetate.

Human polymorphonuclear neutrophilic granulocytes were made adherent by passing them over protein-coated nylon fibre columns and compared with suspended cells for their production of superoxide anion as measured by cytochrome C reduction. The cells were stimulated with chemotactic factors, the ionophore A 23187, and the tumour promoter phorbol myristate acetate. There was no increased O2-. production by adherent cells in the absence of a stimulus. Adherent cells produced considerably higher amounts of superoxide than suspended cells when stimulated with formyl-methionyl-leucyl-phenylalanine, ionophore A 23187, C5a, C5adesArg, and the platelet activating factor 1-o-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine. In contrast, stimulation with phorbol myristate acetate did not result in higher superoxide release from adherent than from suspended cells, and leukotriene B4 and a mononuclear cell-derived chemotaxin did not stimulate either cell to release significant amounts of superoxide. It is suggested that the augmented production of oxygen radicals with certain stimuli contributes to inflammatory symptoms in situations involving adherent granulocytes.

Adherence-induced enhancement of the oxidative burst of human neutrophilic granulocytes: effects of the surface coat and of divalent cations.

Neutrophilic granulocytes are capable of adhering to biological or artificial surfaces. In addition to specific interactions between adherence molecules on the cell surface and corresponding structures on other cells or matrix proteins like fibronection or collagen there appeared to be non-specific attachment as well. Adherence augmentation induced by stimulation with chemotactic factors or cytokines was an active process which did not proceed at 4 degrees C and after removal of divalent cations from the medium. Adherence acted as a priming stimulus increasing the amount of superoxide anion and hydrogen peroxide produced in response to stimulation by certain chemotactic factors and cytokines. Adherence priming like priming by GM-CSF was strongly suppressed by chelation of intracellular Ca2+ ions. The two priming mechanisms differed, however, in their requirement for divalent cations in the external medium: whereas Mg2+ suppressed GM-CSF priming, it synergised with Ca2+ in the adherence-augmented oxidative burst.

Stimulation of human neutrophilic granulocytes by two monocyte-derived cytokines.

Human monocytes upon stimulation with bacterial lipopolysaccharide release two cytokines which modulate the functions of neutrophilic granulocytes (PMN), a monocyte-derived chemotaxin (MOC) and tumor necrosis factor (TNF). Both cytokines stimulated the adherence of PMN on protein-coated nylon-fibers. Whereas MOC is one of the four most potent chemoattractants known, TNF was a most powerful inhibitor of PMN chemotactic migration towards several chemotactic factors including MOC. Neither cytokine stimulated the release of superoxide anion (O2-) or hydrogen peroxide (H2O2) from PMN in suspension. However, TNF, but not MOC, caused the release of considerable amounts of H2O2 and O2- from PMN attached to nylon fibers. The two cytokines have similar effects on the adherence, opposing effects on chemotactic migration and different effects on the oxidative burst of PMN.

Modulation of human neutrophilic granulocyte functions by recombinant human tumor necrosis factor and recombinant human lymphotoxin. Promotion of adherence, inhibition of chemotactic migration and superoxide anion release from adherent cells.

Recombinant human tumor necrosis factor (TNF) and recombinant human lymphotoxin (LT) were analyzed for their effects on inflammation-related functions of human polymorphonuclear neutrophilic granulocytes (PMN) in vitro, TNF at a concentration of 10 U/ml (corresponding to 10(-11) mol/l) enhanced PMN adherence to nylon fibres. It strongly inhibited the chemotactic migration of PMN in the Boyden chamber assay towards the chemotactic tripeptide formyl-methionyl-leucyl-phenylalanine (FMLP), C5a, LTB4 and a monocyte-derived chemotaxin (MOC) without affecting random migration and without being chemotactic itself. It did not stimulate superoxide anion (-O2.) production of PMN in suspension. However, it induced considerable -O2. release from PMN that had become adherent on nylon fibres. All these effects were abrogated by prior incubation of the cytokine with polyclonal and monoclonal antibodies against TNF. LT concentrations of 1,000 U/ml or higher were required to observe a moderate inhibition of chemotactic migration towards the above chemotactic factors and to elicit some -O2. production from nylon fibre-adherent PMN. LT did not increase the adherence of PMN to nylon fibres and it was not chemotactic. The results indicate that TNF is a potent modulator of PMN functions.

Assessment of ferrocytochrome C oxidation by hydrogen peroxide.

The reduction of ferricytochrome C is commonly employed for the quantitation of O2-.H2O2 arising from the dismutation of O2- is capable of oxidizing ferrocytochrome C. In order to assess whether this may interfere with O2- quantitation, the amount of H2O2 required for the oxidation of ferrocytochrome C was determined. While H2O2 concentrations below 10(-5) M were ineffective, one half of the reduced cytochrome was oxidized by 5 x 10(-5) M H2O2 within 15 min. H2O2 in the concentration range at which ferrocytochrome C is oxidized is generated upon interaction of hypoxanthine with xanthine oxidase and upon stimulation of human polymorphonuclear neutrophilic granulocytes by phorbol myristate acetate or the phagocytosis of opsonized zymosan. It is suggested that O2- quantitation by cytochrome C reduction is routinely performed in the presence of catalase.

Novel neutrophil chemotactic factor derived from human peripheral blood mononuclear leucocytes.

Human mononuclear leucocytes isolated from the peripheral blood by centrifugation on Ficoll-Hypaque cushions and adherent on plastic petri dishes, produced a chemotactic factor that attracted human neutrophilic granulocytes to the same extent as did optimal concentrations of the complement split product C5a and the leukotriene B4. The active component eluted from a Sephadex G-50 gel filtration column as a single peak with an apparent molecular weight of 10,000. The chemotactic activity was resistant to reductive cleavage of disulfide bonds and heating at 100 degrees C for 30 min but was lost when reduction and heating were combined. Digestion with a proteolytic enzyme eliminated the attractive potential. The data suggest that this is a novel chemotactic peptide. It is conceivable that it has been seen previously and was mistaken for a lymphokine or interleukin 1.

Inhibition of chemotactic migration of human neutrophilic granulocytes by recombinant human granulocyte-macrophage colony-stimulating factor.

Human recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) was analysed for effects on the migration of human neutrophilic granulocytes by the Boyden chamber assay. At concentrations ranging from 0.1 to 10,000 U/ml (or 10(-12) to 10-mol/l) GM-CSF had neither chemokinetic nor chemotactic activity. When added to the cells in the upper compartment of the chamber GM-CSF dose-dependently inhibited the chemotactic migration towards the tripeptide f-Met-Leu-Phe and the complement split product C5a. Chemotaxis towards f-Met-Leu-Phe was inhibited more efficiently by GM-CSF than C5a-induced migration.

Sonography of placental abnormalities and oligohydramnios in women with elevated alpha-fetoprotein levels: comparison with control subjects.

To evaluate the relationship of placental and amniotic fluid findings to elevated maternal serum alpha-fetoprotein (MS-AFP) levels, we compared sonograms made between 18 and 24 weeks gestational age in 76 women with elevated MS-AFP levels with sonograms of a control group. Patients with fetal malformations, incorrect dates, twins, or lack of follow-up were excluded. Overall, 27 (36%) of 76 patients with elevated MS-AFP levels had placental or amniotic fluid abnormalities compared with only three (3%) of 87 control subjects. Significant differences (p less than .01) were noted in the frequency of periplacental hemorrhage (9% vs 0%), intraplacental sonolucencies greater than or equal to 1.5 cm in diameter (18% vs 3%) and moderate or severe oligohydramnios (17% vs 0%). More patients with elevated MS-AFP levels had placenta previa (4%) or placental thickness greater than or equal to 3.5 cm (12%) than did those in the control group (1% and 5%, respectively), although these differences did not reach statistical significance. Seven (26%) of the 27 patients had more than one abnormality. We conclude that placental and/or amniotic fluid abnormalities are frequently shown on sonograms in women who are examined because of elevated MS-AFP levels.

The "genetic sonogram": comparison of the index scoring system with the age-adjusted US risk assessment.

PURPOSE: To compare two ultrasonographic (US) methods for prenatal detection of fetal Down syndrome. MATERIALS AND METHODS: Genetic amniocentesis was successfully performed in 3,303 consecutive women with high-risk pregnancies (mean gestational age, 17.1 weeks). All patients underwent a complete "genetic US" examination prospectively. Risk was assessed by using (a) various modifications of the index scoring system (ISS) and (b) the age-adjusted US risk assessment (AAURA). RESULTS: The prevalence of Down syndrome in this population was 1.6% (53 of 3,303). By using a threshold of at least 2 points to detect trisomy 21, the best ISS had a sensitivity of 45.3%, false-positive rate of 4.9%, likelihood ratio of 9.3, and positive predictive value in the high-risk population in this study of 13.3%. Lowering the threshold to 1 point increased the sensitivity to 60.4% but increased the false-positive rate to 15.8%. Adding points for age increased the sensitivity to 67.9% but increased the false-positive rate to 24.3%. Results of using AAURA to detect trisomy 21 were nearly identical, with a sensitivity of 43.4% and false-positive rate of 4.9% at a 1 in 36 risk threshold and a sensitivity of 69.8% and false-positive rate of 26.1% at a 1 in 200 threshold. Trisomies 18 and 13 were detected with sensitivities of 80.0% and 100.0%, respectively, with either system. CONCLUSION: The modified ISS and AAURA are equivalent in screening for Down syndrome, with detection of approximately half of all trisomy 21 fetuses at a 5% false-positive rate.