Somatic mosaicism of a greater than 1.7-Mb deletion of genomic DNA involving the entire NF1 gene as verified by FISH: further evidence for a contiguous gene syndrome in 17q11.2.

We report on a third case with neurofibromatosis type 1 (NF1) due to mosaicism for a gross deletion in 17q11.2 covering the entire NF1 gene. The deletion was suspected in Giemsa banded chromosomes and was confirmed by fluorescence in situ hybridization using the cosmids CO919 from the 5' region, GO2121 from the central, H10410 from the 3' region of the NF1 gene, and the 1.7-Mb YAC 947G11 spanning the entire 350-kb genomic DNA of the NF1 gene. The deletion was present in 33% of peripheral blood lymphocytes and 58% of fibroblasts. The clinical manifestations in this 6-year-old male patient were especially severe and extended beyond the typical features of NF1. The patient also displayed facial anomalies, severe and early-onset psychomotor retardation, seizures, spasticity, and microcephaly. These features differ from other large-deletion NF1 patients, even nonmosaic cases. The complex phenotype could be explained by the involvement of coding sequences flanking the NF1 gene, thus supporting the existence of a contiguous gene syndrome in 17q11.2.

Comparison of HER2/neu status assessed by quantitative polymerase chain reaction and immunohistochemistry.

We prospectively evaluated a series of 254 breast cancers by quantitative polymerase chain reaction (PCR) and immunohistochemistry using 3 antibodies: HercepTest, CB11, and TAB250. DNA was extracted from a 10-micron tumor section for PCR, and 4-micron serial sections were taken from the same block for immunohistochemistry. The immunohistochemical results were scored using a semiquantitative immunohistochemical system. A positive tumor by immunohistochemistry had a score of 5 or more. The manufacturer's recommended scoring system was used for the HercepTest. Tumors were positive for gene amplification if the ratio of the HER2/neu gene to control gene after normalization was 2 or more. Of 254 cases, 61 showed gene amplification. For immunohistochemistry, 23% of tumors were positive with CB11, 27% with TAB250, and 37% with the HercepTest. Results for each antibody were compared with PCR results. The overall concordance for the HercepTest was 82%, which was significantly lower than that for CB11 (88%) or TAB250 (87%). The specificity for the HercepTest was 80% compared with 90% for TAB250 and 93% for CB11, while the positive predictive value for the HercepTest was 57% compared with 71% and 76% for TAB250 and CB11, respectively.

The importance of routine HIV testing in the incarcerated population: the Rhode Island experience.

Routine HIV testing in the correctional setting offered to all inmates at entry has played an important role in the diagnosis of HIV in Rhode Island. Diagnosis and treatment of HIV in prisons can further public health goals of HIV control, prevention, and education. Routine HIV testing can be incorporated into primary and secondary prevention programs in correctional facilities. In Rhode Island, where HIV testing is routine at entry into the correctional facility, approximately one third of all persons who test positive are identified in the correctional facility. The proportion of males and females testing positive in the correctional facility versus those testing positive in other facilities has shown a gradual decrease, with positive female HIV tests declining more substantially in recent years. Specific groups, such as males, African Americans, and injection drug users continue to be more likely diagnosed in the state correctional facility than in other testing sites. These differences may reflect barriers to health care access that other community initiatives have failed to address.

Penetration of the colon by a ventriculo-peritoneal drain resulting in an intra-cerebral abscess.

A male child was born with internal hydrocephalus due to aqueductal stenosis requiring a ventriculo-peritoneal shunt on the first postnatal day. Subsequently, the hydrocephalus did not subside but increased. Autopsy at the age of 15 months disclosed the peritoneal tip of the catheter located inside the transverse colon, the cerebral tip of the catheter within a huge abscess of the right cerebral hemisphere. Penetration of the intestine by a ventriculo-peritoneal catheter seems to be a rare event, occasionally resulting in early death due ascending infection.

[Tuberous sclerosis with megalocornea and coloboma of the iris (author's transl)]. / Tuberöse Sklerose mit Megalokornea und Iriskolobom

A mentally and physically retarded 4 1/2 year-old boy with epileptic seizures showed a megalocornea on both sides, a coloboma of the iris in the right eye and a white area at the temporal side of the disc in the left eye. At first a coloboma of the disc was suspected. By further controls at the age of 8 years a typical two diopters elevated nodular opaque white mass was seen in place of the white area in the left eye, in addition two flat tumours were also seen. In the right eye with coloboma of the iris there was also a flat area. Radiographically the right kidney showed two ureters with flat calyces. At the age of 8 years symmetrical face naevi occurred only under atropine medication, and showed at the age of 10 years the typical picture of Pringle's tumours.

Mucolipidosis IV: novel mutation and diverse ultrastructural spectrum in the skin.

Mucolipidosis IV, a severe neurologic and ophthalmologic progressive disorder has a clinical range of onset between early childhood and adolescence entailing clinically severe, moderate, and mild forms, all of them majorly affecting Ashkenazi Jewish patients in an autosomal-recessive fashion owing to mutations in the MCOLN1 gene which encodes a transmembrane protein called mucolipin 1. We report on one of two affected siblings, the older brother having died of ML IV at the age of 33 years, the younger recently at the age of 37 years. Biopsied skin disclosed several types of lysosomal residual bodies, membrane-bound vacuoles, avacuolar lamellar bodies resembling membraneous cytoplasmic bodies, and a diverse spectrum of lipopigments which include curvilinear and fingerprint profiles. Contrary to earlier reports, disease-specific lysosomal residual bodies could not be identified in circulating lymphocytes of our patient. Mutation analysis revealed a homozygous novel mutation of a 34 bp deletion and 3 bp insertion in exon 2 of the MCOLN1 gene, perhaps the reason for this unusual clinical and morphological phenotype.

Preceptor workshops: a collaborative model.

Educational programs for nurses are becoming increasingly reliant on clinicians in hospitals and other health care agencies to act as preceptors for their students. Often there is no formal preparation for this role, nor are rewards provided by the educational institutions. This paper describes a collaborative effort between a university and a college program to prepare and reward nurses who work as preceptors with students in their settings.

[Possibilities for false-negative findings in trisomy 21 screening with FISH]. / Möglichkeit falsch-negativer Befunde beim Trisomie 21-Screening mittels FISH.

In approximately 5% of individuals with Down syndrome aneuploidy results from a chromosomal rearrangement. FISH analysis on chromosome metaphases and interphase nuclei of 5 individuals with Down syndrome carrying different types of chromosome 21 translocations demonstrated the diagnostic efficiency of this method. By use of different commercially available chromosome 21 specific probes we were able to show that only the cosmid probe specific for the Down syndrome critical region (DCR) in 22qll gave reliable results for interphase analysis of trisomy 21, while the use of chromosome 21 centromere- or of painting probes carry a high risk of a false-negative diagnosis in translocation trisomy 21.

The role of HER2/neu overexpression/amplification in the progression of ductal carcinoma in situ to invasive carcinoma of the breast.

HER2/neu overexpression/amplification is seen more frequently in ductal carcinoma in situ, particularly high-grade ductal carcinoma in situ (50-60%), than in invasive ductal carcinoma of the breast (25-30%). To date, however, the role of HER2/neu in the progression of in situ to invasive disease has not been clarified. Two hundred fifty-one breast tumors were retrieved from the pathology files at Mount Sinai Hospital. These included 91 cases of ductal carcinoma in situ, 136 cases of invasive ductal carcinomas with associated ductal carcinoma in situ, and 24 cases of pure invasive carcinomas. All cases were reviewed and stained with two monoclonal antibodies to HER2/neu (CB11 and TAB250). Immunohistochemical staining was recorded using a semiquantitative scoring system (1). Representative cases were also investigated using fluorescence in situ hybridization. HER2/neu protein overexpression (defined as immunohistochemical staining with score of >or=5) was seen in 34% of cases of pure ductal carcinoma in situ, 17% of invasive carcinomas with associated ductal carcinoma in situ, and 12.5% of pure invasive carcinomas (P =.01). Sixty percent of cases of high-grade ductal carcinoma in situ showed HER2/neu protein overexpression, versus 29% of high-grade invasive carcinomas with associated ductal carcinoma in situ and 22% of high-grade pure invasive ductal carcinomas (P =.02). The concordance between the immunohistochemical staining in the in situ and invasive components of individual tumors was 90%. Thirty-three cases were also evaluated by fluorescence in situ hybridization and showed concordance between the immunohistochemical results and the degree of gene amplification in 91% of cases, whereas 3 of 33 cases showed HER2/neu gene amplification (HER2/CEP17 = 2.3-3.7) by fluorescence in situ hybridization in the absence of positive immunohistochemical staining. One case showed HER2/neu gene amplification in the associated ductal carcinoma in situ (HER2/CEP17 ratio = 6.5), with no evidence of gene amplification in the invasive tumor (HER2/CEP17 ratio = 1.14). Multiple genetic events are required for the development of an invasive phenotype. The findings from this study suggest that the genetic event of HER2/neu gene amplification/protein overexpression may not play a key role in the progression of ductal carcinoma in situ to invasive carcinoma and that other molecular alterations may be more important in the initiation of invasion in ductal carcinoma of the breast.

Effects of chronic administration of ethanol on platelets from rabbits with diet-induced hypercholesterolemia. Unchanged characteristics and responses to ADP but reduction of enhanced thrombin-induced, TxA2-independent platelet responses.

To investigate the effect on platelet function of the interaction between dietary cholesterol and moderate, chronic doses of ethanol, hypercholesterolemia was induced in rabbits by 8 weeks of administration of a chow diet with added (0.25% wt/wt) cholesterol; during the eighth week, a moderate amount of ethanol (6% in drinking water) was given. Blood alcohol levels were not detectable in ethanol-treated rabbits at the time of exsanguination. Ethanol did not affect plasma cholesterol levels or the cholesterol to phospholipid molar ratio in platelets. Platelet membrane fluidity, which decreased with cholesterol feeding, was not altered further by administration of ethanol. The overall fatty acid composition of platelet phospholipids was not affected by either cholesterol feeding or chronic ethanol intake. Responses of washed platelets stimulated with either ADP or thrombin were studied to determine whether ethanol administration modified platelet functions in hypercholesterolemia. Primary ADP-induced aggregation was not affected by cholesterol feeding or chronic ethanol intake, but thrombin-induced aggregation and secretion of [14C]serotonin from prelabeled platelets, which were enhanced by cholesterol feeding, were diminished by administration of ethanol to hypercholesterolemic rabbits. This reduction in thrombin-induced responses was also observed with aspirin-treated platelets, which cannot form thromboxane A2. Thus, chronic short-term administration of a moderate amount of ethanol inhibited the enhanced responses of platelets from rabbits with diet-induced hypercholesterolemia, via a thrombin-induced, thromboxane A2-independent pathway.

Enhanced collagen-induced responses of platelets from rabbits with diet-induced hypercholesterolemia are due to increased sensitivity to TxA2. Response inhibition by chronic ethanol administration in hypercholesterolemia is due to reduced TxA2 formation.

The effects of dietary cholesterol and chronic administration of moderate amounts of ethanol on collagen-induced platelet responses were investigated. Three groups of rabbits were fed the following diets for 8 weeks: a normal chow diet, a cholesterol-enriched (0.25% wt/wt) chow diet, and a cholesterol-enriched chow diet plus 6% ethanol in the drinking water for the final week of the dietary period. Cholesterol feeding enhanced collagen-induced responses-aggregation, secretion of [14C]serotonin from prelabeled platelets, and thromboxane formation--of suspensions of washed platelets, and chronic ethanol treatment significantly reduced these enhanced responses. These effects are mediated by thromboxane A2 (TxA2) rather than ADP. Experiments with collagen-stimulated platelets in which feedback amplification of TxA2 was blocked with the prostaglandin H2/TxA2 receptor blocker BM 13.177 and experiments with aspirin-treated platelets stimulated with the stable TxA2 mimetic U46619 showed that cholesterol feeding enhanced platelet sensitivity to TxA2 rather than formation of TxA2 by platelets that had interacted with collagen. Without BM 13.177 or aspirin, TxA2 increased the amount of TxA2 formed by feedback amplification. In contrast, decreased responsiveness to collagen by platelets from cholesterol-fed rabbits given ethanol was due to inhibition of TxA2 formation rather than reduced sensitivity to TxA2. Platelets from cholesterol-fed rabbits given ethanol did not develop tolerance to the acute inhibitory effects of ethanol. Our results indicate that administration of moderate amounts of ethanol to cholesterol-fed rabbits inhibits enhanced collagen-induced responses of platelets by a TxA2-dependent pathway that involves reduction of TxA2 formation rather than reduction of platelet responses to TxA2.

[Activity of the creatine kinase isoenzyme CK-BB in the serum of neonates as an indicator of perinatal damage to the central nervous system]. / Die Aktivität des Creatinkinase (EC 2.7.3.2)-Isoenzyms CK-BB im Serum Neugeborener als Indikator einer frühkindlichen Schädigung des zentralen Nervensystems.

By means of the immunoprecipitation method significantly higher activities of creatine kinase BB isoenzyme were measured in the sera of neonates with CNS symptoms than in the sera of healthy or sick neonates without CNS symptoms. The activity of CK-BB inversely correlated with the one-minute Apgar score. These results suggest a leakage of CK-BB from the damaged CNS tissue into the blood circulation. Determination of CK-BB might be helpful in the assessment of perinatal brain damage.

The activity of letrozole in patients with advanced or recurrent endometrial cancer and correlation with biological markers--a study of the National Cancer Institute of Canada Clinical Trials Group.

A multicenter phase II trial was conducted to define the activity of letrozole in postmenopausal women with recurrent or advanced endometrial carcinoma, who had no more than one prior line of progestins and never had chemotherapy (except adjuvant). Archival paraffin-embedded tumor samples were retrieved to determine the expression level of estrogen (ER) and progesterone receptor (PgR), p53, HER-2, bcl-2 and PTEN protein, and phosphorylation status of protein kinase B (PKB/Akt). Thirty-two eligible patients were treated with letrozole at 2.5 mg daily continuously, of whom 10 (31%) had prior progestins. Of the 28 patients evaluated for response, one complete and two partial responses were noted; overall response was 9.4% (95% confidence interval 2-25%). Eleven patients had stable disease for a median duration of 6.7 months (range 3.7-19.3 months). Amongst 22 patients who had tumor blocks available, the proportion showing positive expression of the following markers includes: PgR (86%), ER (86%), PTEN (82%), phosphorylated PKB/Akt (59%), bcl-2 (45%), p53 (32%), and HER-2 (0%). None of these markers correlated with response to letrozole or disease progression. In conclusion, letrozole is well tolerated but has little overall activity in this cohort of women with endometrial cancer.

[Prenatal diagnosis of alpha-1-antitrypsin phenotype. Case record and prognosis in severe alpha-antitrypsin deficiency Pi ZZ (author's transl)]. / Pränataler Ausschluss eines Alpha-1-antitrypsinmangels. Kasuistik und Prognose bei Phänotyp Pi Z.

A mother had a child with cirrhosis of the liver and alpha-1-antitrypsin deficiency. In a subsequent pregnancy the fetal phenotype Pi MZ was detected by isoelectrofocusing in the amniotic fluid. Quantitative assay of alpha-1-antitrypsin gave results in the normal range. Umbilical vein blood analysis confirmed the antenatal findings. In this case it has been possible to rule out the disease before birth. In this context the clinical importance of alpha-1-antitrypsin deficiency is stressed, its frequency in the European and North-American population and the prognosis with phenotype Pi Z.