Plasma level of LDL-cholesterol at diagnosis is a predictor factor of breast tumor progression.

BACKGROUND: Among women, breast cancer (BC) is the leading cancer and the most common cause of cancer-related death between 30 and 69 years. Although lifestyle and diet are considered to have a role in global BC incidence pattern, the specific influence of dyslipidemia in BC onset and progression is not yet completely understood. METHODS: Fasting lipid profile (total cholesterol, LDL-C, HDL-C, and triglycerides) was prospectively assessed in 244 women with BC who were enrolled according to pre-set inclusion criteria: diagnosis of non-hereditary invasive ductal carcinoma; selection for surgery as first treatment, and no history of treatment with lipid-lowering or anti-diabetic drugs in the previous year. Pathological and clinical follow-up data were recorded for further inclusion in the statistical analysis. RESULTS: Univariate associations show that BC patients with higher levels of LDL-C at diagnosis have tumors that are larger, with higher differentiation grade, higher proliferative rate (assessed by Ki67 immunostaining), are more frequently Her2-neu positive and are diagnosed in more advanced stages. Cox regression model for disease-free survival (DFS), adjusted to tumor T and N stages of TNM classification, and immunohistochemical subtypes, revealed that high LDL-C at diagnosis is associated with poor DFS. At 25 months of follow up, DFS is 12% higher in BC patients within the third LDL-C tertile compared to those in the first tertile. CONCLUSIONS: This is a prospective study where LDL-C levels, at diagnosis, emerge as a prognostic factor; and this parameter can be useful in the identification and follow-up of high-risk groups. Our results further support a possible role for systemic cholesterol in BC progression and show that cholesterol metabolism may be an important therapeutic target in BC patients.

Partial characterization of cytoplasmic compartments involved in the endocytic process of Tritrichomonas foetus.

The endocytic pathway of Tritrichomonas foetus, a parasitic protozoan of cattle, was studied using (a) vital dyes, such as Lucifer yellow, neutral red and acridine orange, (b) cationized ferritin, (c) gold-labeled lactoferrin and lectins: HPA, UEA, PNA and LPA, and (d) DAMP (3-(2,4-dinitroanilino) 3' amino-N-methyldipropylamine). Light and confocal laser microscopy as well as transmission electron microscopy were used in this study. Assays were monitored by fluorescence and electron microscopy after exposing the parasites to different conditions. Cells that were incubated at 15 degrees C or 20 degrees C with gold-labeled lactoferrin and processed for electron microscopy show that of 15 degrees C this ligand is found only in an early endosomal compartment and at 20 degrees C it is found in late endosomes but not in lysosomes. Immunocytochemical data from cryosections using DAMP as a pH probe show that T. foetus has acidic compartments, with a pH range of 5.2 to 6.6, with variable morphology, localization and size. Lectin-binding sites and anionic sites were also internalized and appear to be associated with membranes lining the vacuoles. Images of patching and shedding of these sites were also observed when HPA and UEA were used.

The DNA puff BhB10-1 gene encodes a glycine-rich protein secreted by the late stage larval salivary glands of Bradysia hygida.

We present the molecular characterization of a gene of Bradysia hygida DNA puff B10 whose temporal expression in the salivary gland correlates with the puff expansion. The transcription unit of this gene, named BhB10-1, was mapped in a 2-kb EcoRI genomic fragment that is amplified in the salivary gland of late fourth instar larvae. Its 1.3-kb transcript undergoes poly-A tail shortening during development, indicating that post-transcriptional controls as well as transcription activation are involved in the temporal regulation of the BhB10-1 gene. Analysis of the deduced amino acid sequence from the cDNA indicates that the BhB10-1 protein is a glycine-rich secretory protein. A BhB10-1-fusion protein expressed in bacteria was used to raise polyclonal antibodies. Using an immunopurified antibody, we identified the product of the DNA puff BhB10-1 gene as a 23-kDa polypeptide that is produced mainly by the salivary gland regions S1 and S3 and is present in the saliva of late larvae. This is the first direct identification of a protein encoded by a DNA puff amplified gene.

RAPADILINO syndrome.

We report on a boy with severe radial hypoplasia, absent thumbs and patellae, short stature, persistent diarrhea, slender nose and normal intelligence as another example of the RAPADILINO syndrome.

CODAS syndrome: a new distinct MCA/MR syndrome with radiological changes of spondyloepiphyseal dysplasia. Another case report.

We report on a young girl with psychomotor delay, cataracts, abnormally shaped teeth, malformed ears, and radiological findings of spondylo-epiphyseal dysplasia. The clinical picture resembles the CODAS syndrome described by Shebib et al. [Am J Med Genet 40: 88-93, 1991].

Absent tibiae--polydactyly--triphalangeal thumbs with fibular dimelia: variable expression of the Werner (McKusick 188770) syndrome?

We report on a family in which the autosomal dominant Werner syndrome (WS) (MIM# 188770) affects ten members in three generations. Besides the absent tibiae the propositus had duplication of the fibulae. Possible pathogenetic mechanism is discussed.

Tetrasomy 9p caused by idic (9) (pter----q13----pter).

Cytogenetic investigation on a malformed male infant showed an extra chromosome similar to chromosome 9 in all metaphases studied. GTG, CBG, and G-11 staining suggested that the extra chromosome was an abnormal 9, permitting the identification of the chromosome constitution as 47,XY,+idic (9) (pter----q13----pter).

Autosomal dominant osteosclerosis type Stanescu: the third family.

We describe a family with Stanescu osteosclerosis. The propositus and his mother were short and had cortical sclerosis of the long bones, deficient facial sinus development, cranial bone malformations, and normal intelligence. To the best of our knowledge, only two such families have been described previously. The autosomal dominant pattern of inheritance of this skeletal dysplasia is reinforced, as there are many other reportedly affected relatives, including the maternal grandfather, uncles, and aunts of the propositus. The findings of wormian bones and calcification of the falx, not previously described, may be added to the phenotype.

Limb deficiency with or without Möbius sequence in seven Brazilian children associated with misoprostol use in the first trimester of pregnancy.

Misoprostol, a synthetic analog of prostaglandin, has been widely used in Brazil as an abortifacient. Abortion is illegal in Brazil. An uncertain number of these abortion attempts are unsuccessful and the pregnancy continues. We report on 7 patients whose mothers attempted to abort using this drug in the first trimester of gestation without success. The 7 patients presented with limb defects and in 4 of them a diagnosis of Möbius sequence was made.

A dual role of 20-hydroxyecdysone in the control of protein synthesis related to DNA puff activity in the anterior region of Bradysia hygida (Diptera, Sciaridae) salivary gland.

During the last 30 h of the larval stage, the salivary glands of Bradysia hygida show the amplification of some genes, resulting in the formation of two successive groups of DNA puffs, which direct the synthesis of two different sets of polypeptides. Incubation of anterior (S1) salivary gland regions, at age E7, beginning of first group of DNA puffs activity, in culture medium for 2 to 10 h results in a decrease in the synthesis of the polypeptides characteristic of this period. However, during subsequent incubation (from E7 to E7+12 h-24 h), when the second group of DNA puffs is active, S1 regions were able to synthesize some polypeptides characteristic of this period. The role of 20-OH ecdysone was studied, in vitro and in vivo, during these two periods of protein synthesis in S1 regions. The presence of the hormone was shown to be necessary to maintain, in vitro, the synthesis of the first set of polypeptides and was strongly inhibitory, in vitro and in vivo, to the synthesis of the second set of polypeptides. Thus, it is likely that the activity of the two distinct groups of DNA puffs is under opposite 20-OH-ecdysone control mechanisms.

Non-goblet cell population of Barrett's esophagus: an immunohistochemical demonstration of intestinal differentiation.

Barrett's esophagus develops with the following 2 distinct types of lining mucosa: with and without specialized intestinal metaplasia (SIM). Goblet cells found only in SIM areas identify an intestinal phenotype, recognized as the histological hallmark diagnosing Barrett's metaplasia, and selecting high-risk patients for endoscopic surveillance. The columnar non-goblet cells are the major component of the heterogeneous Barrett's metaplastic cell population and are present in areas either with or without SIM. Their significance in the differentiation of columnar-lined esophagus, and their relationship to malignancy, is still unclear. This immunohistochemical study used two markers of enterocytic differentiation, to explore the intestinal phenotype of the non-goblet cell population of Barrett's epithelium and Barrett's-associated adenocarcinoma cells. Sucrase-isomaltase (SI) and dipeptidilpeptidase IV (DPP) immunoexpression was assessed in paraffin-embedded samples of 12 surgical specimens containing Barrett's esophageal mucosa in association with adenocarcinoma/high grade dysplasia. Ileal mucosa and mucosa from normal gastric and esophageal segments of the surgical specimen were used as positive and negative controls, respectively. SI and DPP were expressed by the neoplastic cells and the columnar non-goblet, being negative in goblet cells. The localization of the enzymes was predominantly apical for SI and cytoplasmatic for DPP. There was immunoreactivity for SI in 58.3% of the carcinomas and in 66.6% of Barrett's mucosa, with equal frequency in areas with and without SIM. DPP was identified in 66.6% of the carcinomas, in 50% of the cases of Barrett's metaplasia with SIM, and in 75% of those without SIM. The columnar non-goblet cell components of Barrett's metaplasia contain small intestine enzymes in the areas either with or without SIM, which suggests that they identify an "incomplete form" of intestinal metaplasia. The demonstration that the two enzymes, SI and DPP, are produced by the columnar non-goblet cell metaplastic population and by the neoplastic cells of the associated adenocarcinoma, indicates that, in addition to the goblet cells, the non-goblet elements may also be involved in the malignant transformation of Barrett's esophagus.

The control of BhB10-1 gene expression in the salivary gland of Bradysia hygida (Diptera, Sciaridae) is disrupted in vivo by a delayed effect of cycloheximide.

BhB10-1 is an amplified gene present in DNA puff B10. This gene is very active in the salivary gland regions S1 and S3 at the end of the larval development. Two transcripts of this gene, 1.3 and 1.1 kb in size, were detected. A secretory protein, SP23, is the product of BhB10-1. In this work, we present evidence supporting the hypothesis that a biphasic process of mRNA degradation is an important component in the control of BhB10-1 gene expression. The 1.3 kb transcript, by a process of poly(A) tail shortening, is converted to the inactive transcript of 1.1 kb which is detected during and after the period of SP23 expression. Cycloheximide in very low concentration, if applied at a proper time, can disrupt this process leading to extended periods of 1.3 kb RNA detection and SP23 synthesis. A tentative model is proposed to explain this phenomenon.

Does the new TNM classification (1997) improve prognostic stratification in gastric cancer submitted to R0 surgery?

AIMS: To determine the eventual advantage of the new 1997 TNM as prognosis predictor for gastric cancer patients submitted to an R0 resection and to compare it with two other lymph-node involvement classifications, the 1990 TNM and the Okusa system. METHODS: From January 1980 to December 1995, an R0 resection was performed as primary therapy in 275 cases of gastric cancer. These operations consisted of a total or sub-total gastrectomy and of a D2 type lymph-node dissection. Tumour classification was performed according to 1990 and 1997 TNM systems, and to the Okusa lymph node classification. The statistical methods used to evaluate prognostic value were: Kaplan-Meier survival estimates; the log-rank test for univariate analysis; and Cox's model for multivariate analysis. RESULTS: The 1990 TNM showed the best stratification power in univariate analysis. In multivariate analysis, the Okusa classification was identified as the best prognostic index (P<0.01). The 1997 TNM showed worse stratification capability than the two other systems. CONCLUSIONS: In the present series, the new TNM (1997) did not improve the prognostic stratification of lymph-node involvement. An adequate and universal system for lymph-node stratification is necessary and further validation of these classifications is needed.

Immunoscintigraphy with 99mTc-labelled anti-CEA monoclonal antibody in colorectal carcinoma.

With the introduction of immunoscintigraphy (IS) with 99mTc-labelled anti-CEA monoclonal antibodies (MoAb), a clinical relevant method in nuclear medicine can be expected in the diagnosis and follow up of colorectal cancers. We performed IS (whole body, planar and SPECT) with a 99mTc-labelled intact anti-CEA MoAb (BW 431/26) in 18 patients with primary colorectal carcinoma, metastases or suspicious recurrences from colorectal carcinoma. The results of anti-CEA IS, serum CEA and Ca 19-9 levels were evaluated. Immunoscintigraphy yielded an overall sensitivity of 70.0%, 37.5% for primary tumors, 75.0% for recurrences and 100% for distant metastases. Serum CEA levels were elevated in 10 out of 18 patients (sensitivity 55.5%) and Ca 19-9 were elevated in eight out of 18 patients (sensitivity 44.4%). In the group of patients with metastases, CEA had a sensitivity of 100% and Ca 19-9, of 83.3%. From this prospective study, we can conclude that IS with 99mTC-BW 431/26 is a reliable tool in the post-operative follow-up study of patients with colorectal carcinoma, namely in the detection of distant metastases.

Enterocytic columnar non-goblet cells of Barrett's esophagus--an immunohistochemical demonstration of association with malignant evolution.

Barrett's epithelium (BE), the esophageal columnar-lining with intestinal differentiation, is a premalignant condition predisposing to adenocarcinoma. Columnar cells are the prevalent element of BE, but the hallmark of intestinal differentiation is the goblet population that defines the specialised columnar epithelium (SCE). We have demonstrated that columnar cells adjacent to Barrett's adenocarcinoma (BA) exhibit enterocytic features in areas with and without SCE. Nevertheless, the relationship between malignancy and the presence of these elements is not established. To investigate whether intestinal differentiated cells, other than goblet cells, are associated to neoplasia we compared the prevalence of enterocytic features in columnar elements with and without associated BA through the use of sucrase-isomaltase (SI) immunoreactivity in 31 columnar esophageal segments (CLES) and 12 BA. In metaplasia, SI was only expressed at the columnar cells. Apical staining was exclusive of CLES with SCE. SI was present at the cytoplasm in 22.2% of CLES without SCE. Apical SI occurred in BE with and without carcinoma, similarly in areas with and without SCE (p = 0.11 and p = 0.50, respectively). In areas with SCE, columnar cells with apical SI were more frequent in cases of BE adjacent to carcinoma than in cases without neoplasia but the difference did not reach significance (p = 0.053). In areas without SCE, apical SI was significantly (p = 0.01) more frequent in cases with carcinoma. Apical SI was equally found in neoplastic as in metaplastic areas, with and without SCE, (p = 0.07 and p = 0.40, respectively). In conclusion this study on the frequency of SI on CLES with and without neoplasia demonstrated that additionally to SCE, metaplastic enterocytic cells are also associated with malignancy. It also confirmed that the presence of intestinal features are underestimated if only goblet elements are used for its identification, reinforcing the utility of the immunohistochemical recognition of enterocytic characteristics for establishing the diagnosis of BE.

A 28-fold increase in secretory protein synthesis is associated with DNA puff activity in the salivary gland of Bradysia hygida (Diptera, Sciaridae).

When the first group of DNA puffs is active in the salivary gland regions S1 and S3 of Bradysia hygida larvae, there is a large increase in the production and secretion of new salivary proteins demonstrable by [3H]-Leu incorporation. The present study shows that protein separation by SDS-PAGE and detection by fluorography demonstrated that these polypeptides range in molecular mass from about 23 to 100 kDa. Furthermore, these proteins were synthesized mainly in the S1 and S3 salivary gland regions where the DNA puffs C7, C5, C4 and B10 are conspicuous, while in the S2 region protein synthesis was very low. Others have shown that the extent of amplification for DNA sequences that code for mRNA in the DNA puffs C4 and B10 was about 22 and 10 times, respectively. The present data for this group of DNA puffs are consistent with the proposition that gene amplification is necessary to provide some cells with additional gene copies for the production of massive amounts of proteins within a short period of time.

Cloning of a B10 DNA puff sequence developmentally amplified and expressed in the salivary gland of Bradysia hygida.

A recombinant clone carrying a 2-kb fragment was isolated from a mini-library of the B10 DNA puff of Bradysia hygida. This fragment was amplified in the salivary gland during the period of DNA puff formation. Amplification started when DNA puff anlage was formed and continued to increase, reaching a maximum of about 10-fold 28 h later. Northern blot hybridization experiments showed that this 2-kb fragment was complementary to two RNA species of about 1.3 kb and 1.1 kb, which are developmentally regulated in the salivary gland. Maximum amounts of these messages were present when the B10 puff is fully expanded.

[Abdominal versus thoracic approach for myotomy in esophageal achalasia]. / Via abdominal versus torácica para a miotomia por acalásia do esófago.

Miotomy is the most common operation for the treatment of achalasia of the esophagus. The most important complication of this operation is gastro-esophageal reflux, and controversy exists in the choice of a thoracic or abdominal approach to this operation. From 1974 until 1988, our group performed 45 miotomies for achalasia of the esophagus. Follow-up was obtained in 82% of the patients. The thoracic approach was used in 21 cases (tor), and 24 patients were operated through an abdominal approach (abd). All the miotomies had an anti-reflux procedure associated. Post-operative evaluation consisted of: clinical score determination; endoscopy with biopsy; manometry; 24 Phmetry. The clinical score revealed that 47% of the patients in the group tor were asymptomatic, and in the group abd this rate was 53%. Absence of macroscopic esophagitis was registered in 75% of the patients in the group tor, and in 91% in the group abd. Microscopic esophagitis was observed in 42% of the cases in the group tor, and in 45% of the group abd. Manometry revealed a low pressure in the inferior esophageal sphincter in 90% of the cases of the group tor, and in 80% of the group abd. Pathological gastro-esophageal reflux, detected by 24 h Phmetry, was present in 29% of the group tor, and in 36% of the group abd. The results in these two groups were very similar, uniformly favourable, and without evidence of superiority of either operation. The choice of a thoracic, or abdominal approach should depend on associated factors such as patient age, respiratory disease, etc.

[Palliative surgery in carcinoma of the stomach. Retrospective study of 112 consecutive cases]. / Cirurgia paliativa por carcinoma do estômago. Estudo retrospectivo de 112 casos consecutivos.

From January 1980 until October 1991 we operated 112 patients with Gastric cancer in which Surgery was considered palliative by intra-operative criteria or by pathological analysis of the resected specimen. Locally irresectable tumour was found in 24.1% of the cases, peritoneal metastases (mets.) in 21.4%, liver mets. in 17%, lymphatic mets. in 16.1%, and other mets. in 21.4%. Resections were possible in 57 patients (50.8%), with a mortality rate of 10.5%, which was similar to the mortality in the non-resection group (7%) p = 0.2. Median survival for the entire group was 7 months, the non-resection group had a median survival of 4 months, and the resection group of 18 months (p = 6.480 e-0.7). Locally advanced tumors had a better outcome than the metastatic group (p = 0.05), but no difference was observed between patients with liver or peritoneal mets. Patients in stage 3 and 4 of the disease had a different prognosis (p = 0.03), and the resection group within each stage fared better.

[The curative surgery of periampullary tumors. The results of 48 resections]. / Cirurgia de intenção curativa nos tumores peri-ampulares. Resultados de 48 ressecções consecutivas.

Periampullary tumors form a clinical entity with common symptoms, similar therapeutic options, unsatisfactory resectability rates and unfavorable prognosis. From April 1970 until March 1994, one hundred and twenty-seven patients with periampullary carcinoma were operated by our surgical team. In 48 of these patients, a resection for cure was performed (38%). Resectability rates varied according to the origin of these tumors, i.e., pancreas-20%, ampulla-76%; distal bile duct-71%, periampullary duodenum-88%. Pancreatic tumors showed a different resectability rate from the other periampullary carcinomas (p = 0.04). Forty-two of these patients had a pancreatoduodenectomy and in the remaining 6 cases a total pancreatectomy was performed. Fifteen patients had major post-operative morbidity (31%) and 8 of these cases died in-hospital (17%). Follow-up data was available in 81% of the patients, survival estimates were calculated according to the Kaplan-Meier method and survival comparisons were made with the Log-rank test. Median survival for resected pancreatic carcinoma was 6 months and for resected tumors of the ampulla 37 months. In this group of patients, pancreatic tumors showed a different survival rate from the remaining periampullary tumors (Log-rank-p = 0.002). This work evidences the need to improve management of periampullary tumors, particularly in-hospital mortality and long-term survival. To achieve these goals, patients with periampullary tumors should be treated in specialized centers and research to improve local and systemic control of this disease should be pursued.