Detection of pS2 messenger RNA in gynecological cancers.

Estrogen-inducible pS2 mRNA was previously detected in human cancer cell lines the growth of which was sensitive to estrogen. In the present study, the expression of the pS2 gene was analyzed in 111 gynecological carcinomas. The pS2 message was detected in greatest abundance in 6 primary carcinomas of the ovary (6 of 29), 4 of these being mucinous cystadenocarcinomas. A secondary carcinoma of the ovary, and another of the omentum (1 of 4), also contained detectable levels of pS2 mRNA. Weak pS2 mRNA signals were occasionally observed in endometrial (2 of 55) and cervical carcinomas (2 of 33) as well. There was a poor correlation between estrogen receptor and pS2 mRNA in ovarian carcinomas.

Expression of transforming growth factor-beta1 (TGF-beta1) and urokinase-type plasminogen activator (u-PA) genes during arterial repair in the pig.

OBJECTIVE: The transition from quiescence to proliferation in vitro is accompanied by early expression of proliferation-associated genes encoding products including cytokines and enzymes. We aimed to investigate TGF-beta1, u-PA and PAI-1 gene expressions in relation to proliferation and extracellular matrix (ECM) protein gene expressions in porcine arteries following injury. METHODS: Right iliac arteries of juvenile pigs were de-endothelialised and harvested at fixed times after injury. RNA was then extracted and analysed by Northern blot analysis. RNA transcripts in thickened neointima of arteries were examined by in situ hybridisation using digoxygenin-labelled cDNA probes. RESULTS: TGF-beta1, u-PA and PAI-1 transcripts were rapidly elevated (2-8h) and preceded a peak in histone mRNA at 24h after arterial injury. A second prolonged rise in TGF-beta1 mRNA at 4d coincided with elevated ECM protein gene expression. TGF-beta1 gene expression was detected in neointimal cells lining the arterial lumen at 4wk after injury. CONCLUSIONS: The timings of increases in TGF-beta1, u-PA and PAI-1 mRNAs in injured arteries are consistent with contributions to processes prior to proliferation. The observation of a second protracted elevation in TGF-beta1 expression is supportive of an additional role in stimulation of ECM protein synthesis. Functional specialisation exists within the thickened intima of arteries late in repair.

Detection of mRNA for carbonic anhydrase II in human osteoclast-like cells by in situ hybridization.

Carbonic anhydrase II (CA II) plays an important role during osteoclastic bone resorption. Biochemical investigations of gene expression of CA II, however, have been hampered by difficulty in obtaining sufficient numbers of purified osteoclasts. In this study, we describe a nonradioactive, digoxigenin-labeled cDNA in situ hybridization technique capable of determining the pattern of CA II gene expression in human osteoclast-like cells (OC-like cells) at the single-cell level. The results showed that CA II mRNA was located in the cytoplasm of both imprinted and cultured OC-like cells from a giant cell tumor of bone. On the other hand, no evidence of CA II mRNA was found in either the mononuclear cells (tumor cells) of giant cell tumor of bone or osteosarcoma cells. There is a significant correlation between in situ hybridization and northern blot analysis for CA II mRNA in both the giant cell tumor of bone and the osteosarcoma. Our results also indicated that quantitation of in situ hybridization can be achieved by computed cytophotometry.

Cloning, sequencing, and functional characterization of the rat homologue of receptor activator of NF-kappaB ligand.

A complementary DNA (cDNA) encoding the rat homologue of receptor activator of NF-kappaB ligand/osteoprotegerin ligand/osteoclast differentiation factor/tumor necrosis factor (TNF)-related activation-induced cytokine (RANKL/OPGL/ODF/TRANCE) was cloned and sequenced from tibias of ovariectomized (OVX) rats. The predicted amino acid sequence of rat RANKL (rRANKL) has 84% and 96% identity to that of human and mouse RANKL, respectively, and 35% and 37% similarity to that of human and mouse TNF-related apoptosis-inducing ligand (TRAIL), respectively. RANKL transcripts were expressed abundantly in the thymus and bone tissues of OVX rats. rRANKL has a single hydrophobic region between residues 53 and 69, which is most likely to serve as a transmembrane domain. The long C-terminal region containing beta-sheet-forming sequences of the TNF-like core is considered the extracellular region. Three truncated domains within the TNF-like core region were expressed as glutathione S-transferase (GST) fusion proteins and investigated for their ability to induce osteoclastogenesis. The results showed that GST-rRANKL (aa160-318) containing the full TNF-like core region had the highest capability to induce the formation of osteoclast-like cells from RAW264.7 cells. GST-rRANKL (aa239-318 and aa160-268) had lesser degrees of osteoclast inductivity. Furthermore, the GST-rRANKL (aa160-318) is capable of (1) inducing osteoclast formation from rat spleen cells in the presence of macrophage colony-stimulating factor (M-CSF), (2) stimulating mature rat osteoclast polarization and bone resorption ex vivo, and (3) inducing systemic hypercalcemia in vivo; thus the full TNF-like core region of rRANKL is an important regulator of calcium homeostasis and osteoclastic function.

Prognostic significance of pS2 mRNA in breast cancer.

The oestrogen-inducible pS2 protein has previously been associated with good prognosis for breast cancer patients. In 1987-1988 a series of 145 primary breast cancers were examined for pS2 mRNA using northern blots. On recent examination of mortality data, we were unable to find any association between tumour pS2 positivity and patient survival. One patient in 6 died within 5 years of surgery, regardless of pS2 status. In the oestrogen receptor positive/progesterone receptor positive tumour subgroup of patients, we found no evidence of increased survival for pS2-positive tumours. These results do not support use of pS2 as an indicator of increased survival in an average breast cancer patient population.

The p53 tumor-suppressor gene is overexpressed but not mutated in human atherosclerotic tissue.

Excessive proliferation of smooth muscle cells (SMC) in the arterial intima is thought to be a major contributing factor to the development of atherosclerosis. One of the theories put forward to explain the aberrant growth is that genetic alterations similar to those observed in cancers also occur in the SMC. Since mutations in the p53 tumour suppressor gene and the c-Ki-ras oncogene are the most common genetic alterations observed in a wide variety of tumour types, we searched for evidence of mutation in human atherosclerotic tissue. DNA was extracted from atherosclerotic plaques and screened for p53 and c-Ki-ras gene mutations using polymerase chain reaction-single strand conformation polymorphism analysis. Tissue sections were also examined for overexpression of p53 protein using an immunohistochemical technique. The molecular analysis showed that wild-type p53 and c-Ki-ms gene sequences were present in all 54 specimens examined. Overexpression of p53 protein was found in 61% of samples. Our results do not support the view that genetic alterations similar to those occurring in cancer contribute to the abnormal proliferation of SMC, although we caution that only two cancer-related genes were studied. Epigenetic changes to the gene products could play a role however, as evidenced by overexpression of p53 protein in many of the atherosclerotic specimens.

The reliability of highly elevated CA 19-9 levels.

CA 19-9 is used as a tumour marker of the upper gastrointestinal tract. However, extremely elevated CA 19-9 levels are found also in patients with benign diseases. Cholestasis was present in 97.1% of patients with high elevated CA 19-9, independent of their primary disease. 50% of patients with non-malignant diseases and increased CA 19-9 levels showed liver cirrhosis, cholecystitis, pancreatitis and/or hepatitis. In 8.8% no explanation was found for the extremely high CA 19-9 level. The results provide evidence of different factors influencing the CA 19-9 level.

Gene expression of vascular endothelial growth factor in giant cell tumors of bone.

The production of vascular endothelial growth factors (VEGF), a major cause of neoangiogenesis, is a prerequisite for tumor growth and invasion. VEGF have also been shown to be important for the formation of osteoclasts. Because giant cell tumors of bone (GCT) are frequently hypervascular and have the ability to recruit macrophages and multinucleated osteoclast-like giant cells, we evaluated the levels of VEGF gene transcript in several of these tumors using Northern blot analyses, semiquantitative reverse transcription polymerase chain reaction (RT-PCR), fluorescence in situ hybridization (FISH), and immunohistochemistry. Our results showed that three major isoforms of VEGF (121, 165, and 189) were expressed in all cases of GCT investigated, with isoform 121 transcripts the most abundant. By both FISH and immunohistochemistry, we have shown that VEGF was present in spindle-shaped stromal-like tumor cells, round macrophage-like cells, and osteoclast-like multinucleate giant cells. Moreover, we have shown that the levels of VEGF gene expression but not microvessel density correlated with Enneking's clinical stage of GCT. There were higher levels of VEGF gene expression in stage III GCT than in stage I/II GCT (P < .0357). In conclusion, our results indicate that overexpression of VEGF may be associated with the advanced stage of the neoplasm.

Common infections in the history of cancer patients and controls.

The association between the frequency of manifest infectious diseases and cancer risk was investigated in a case-control study at Heidelberg, FRG. A total of 255 cases with carcinomas of the stomach, colon, rectum, breast, and ovary, as well as 255 population controls and 230 hospital controls were interviewed using a standard questionnaire. Controls were matched to the cases for age, sex, and region of residence at the time of the interview. A history of common colds or gastroenteric influenza prior to the interview was found to be associated with a decreased cancer risk. Thus the odds ratios for "three or more common colds per year (on average)" versus "no common cold within the last 5 years prior to the interview" were 0.18 (95% CI = 0.05-0.69) and 0.23 (95% CI = 0.06-0.89) relative to population controls and hospital controls, respectively. There was no apparent relationship between childhood infections or other diseases reported in the earlier history, and cancer risk. While the findings are supported by previous studies and fit well into the results of other fields of cancer research, a conclusive interpretation and biological explanation cannot yet be given.

Severe abdominal pain and thrombocytopenia--typical symptoms of occult jejunal diverticulum perforation?

Complicated small-bowel diverticula cause abdominal pain, gastrointestinal hemorrhage, small-bowel obstruction, and peritonitis. The present patient, had an occult perforation of a small-bowel diverticulum. There were diverticula throughout the whole small bowel. Preoperatively thrombocytopenia (98,000 thrombocytes/cc), was noted. Without any special treatment, i.e., transfusion, the thrombocyte level increased after surgical treatment to normal levels. Although the incidence of small-bowel diverticula appears to be low (0.1%-2.3%) complications may become life-threatening. The level of thrombocytopenia may reflect the extent of inflammation.

3-Methylcrotonylglycine excretion in 3-hydroxy-3-methylglutaric aciduria.

1. 3-Methylcrotonylglycine was identified in urine from an infant with 3-hydroxy-3-methylglutaric aciduria. 2. The concentration of 3-methylcrotonylglycine in urine was approximately one sixth of that of the other metabolite of 3-methylcrotonyl-CoA, 3-hydroxyisovaleric acid. 3. The presence of both metabolites in the infant's urine indicates an inhibition of 3-methylcrotonyl-CoA carboxylase activity in tissues of the infant.

3-Hydroxy-3-methylglutaric aciduria: deficiency of 3-hydroxy-3-methylglutaryl coenzyme A lyase.

A marked deficiency of 3-hydroxy-3-methylglutaryl coenzyme A lyase activity is present in cultured skin fibroblasts from a baby with 3-hydroxy-3-methylglutaric aciduria.

Simultaneous assessment of fetal lung maturity and sebaceous gland function by gas chromatography.

Lipid profiles were analysed by gas chromatography of underivatised extracts from pellets and supernatant fractions of amniotic fluid (AFP and AFS). Peaks, identified as diglycerides resulting from on column decomposition of phosphoglycerides at high temperature, were obvious in gas chromatograms of AFS. The C32 diglyceride, mainly derived from dipalmityl lecithin, was not a prominent peak in AFS from pregnancies from which babies subsequently developed respiratory distress syndrome. There was evidence of a moderate increase of squalene in AFS and AFP for 10 pregnancies of 36-38 weeks gestation resulting in the birth of small for gestational age babies. The increase was not obvious for 21 out of 22 control pregnancies delivering appropriate for gestational age babies. This demonstrated that we had an assessment of two fetal functions in one chromatographic run, the ratio of C32/C34 diglycerides providing a measure of lung maturity and squalene providing an index of proximity to term or of the presence of intrauterine growth retardation.

Studies on a child suspected of having a dficiency in 3-hydroxy-3-methylglutaryl-Co A lyase.

The urine of a child who presented with hyperammonemia was found to contain elevated levels of 3-hydroxy-3-methylglutaric acid, 3-methylglutaconic acid and 3-methylglutaric acid. An increased excretion of these organic acids has been reported previously in a child with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency. Enzyme studies using cultured fibroblasts from this patient, however, indicated that the 3-hydroxy-3-methylglutaryl-CoA lyase activity was not markedly reduced.

Prenatal diagnosis of X-linked ichthyosis.

Prenatal diagnosis of X-linked ichthyosis in a case of steroid sulfatase deficiency was made at 16 weeks by the demonstration of (1) high levels of dehydroepiandrosterone sulfate in amniotic fluid; (2) gross deficiency of steroid sulfatase activity in cultured amniotic fluid cells; (3) very low estriol concentrations in maternal blood and urine; (4) increased maternal plasma dehydroepiandrosterone sulfate; and (5) a characteristic maternal urinary steroid profile with greatly increased levels of 16 alpha-hydroxydehydroepiandrosterone. The latter method is particularly useful since it requires no invasive procedures for the patient and is very specific.

3-Hydroxy-3-methylglutaric aciduria, combined with 3-methylglutaconic aciduria.

1. A baby with severe metabolic acidosis was found to excrete abnormal amounts of 3-methylcrotonic acid, 3-methylglutaconic acid and 3-hydroxy-3-methylglutaric acid in urine. 2. Several other abnormal constitutents appear to be products of side-reactions and include 3-hydroxy-3-methylbutyric acid and 3-methylglutaric acid. 3. The profile of acids in urine indicates a blockage in the sixth step of leucine catabolism, the cleavage of 3-hydroxy-3-methylglutaryl-CoA to acetoacetic acid and acetyl-CoA.

Hormone-sensitive gene expression in breast tumours.

The expression of genes specifically induced by estrogens (pS2), prolactin (PIP) or progestins (Pg8) was measured in primary breast tumours. A highly augmented pS2 gene expression was evident in 55% of estrogen receptor (ER)+, progesterone receptor (PR)+ tumours but was absent in ER- PR- tumours. There was no clear cut correlation between augmented levels of PIP and Pg8 mRNAs in tumours and ER and PR status. Tumours from premenopausal patients were more likely to contain high levels of Pg8 mRNA (P less than 0.038), whereas tumours from postmenopausal patients tended to have augmented levels of PIP mRNA (P less than 0.053).

Clinician/patient interaction: communicating the benefits and risks of oral contraceptives.

Myths and misperceptions continue to influence women's opinions about oral contraceptives (OC), despite the immense body of evidence regarding OC safety and efficacy. Patient opinions about OC failure rates and health risks are often far from proven fact, and the health benefits of OC are too often unrecognized. Because successful OC use requires an informed patient, effective communication between clinicians and their patients is needed to correct misinformation, relieve unnecessary fears, and increase OC use. A variety of interactive counseling skills and attitudes can improve the process of patient counseling; they are especially crucial for adolescents, who may be reluctant or unable to easily articulate their concerns. Counseling messages can be geared specifically to the interests and concerns of particular age groups: adolescents, young adults, and perimenopausal women. With thoughtful planning and involvement of all members of the healthcare team, clinicians can create time and opportunities for the provision of consistent, appropriate counseling to all candidates for OC use.

The role of vitamin D3 in the aetiology of abdominal aortic aneurysms.

Abdominal aortic aneurysms (AAAs) have traditionally been attributed to atherosclerosis, although there is increasing epidemiological, biochemical and genetic evidence that aneurysmal arterial disease is different from occlusive atherosclerosis. One of the most consistent biochemical findings in the aneurysmal aorta is a significant reduction in elastin protein; the cause, for this remains unclear. There is in vitro evidence that vitamin D3 (1,25 dihydrocholecalciferol) inhibits the production of elastin by smooth muscle cells. On the basis of this observation and the possibility that some subjects may be exposed to excess vitamin D3, the hypothesis that vitamin D3 may be a previously unrecognized aetiological factor in the pathogenesis of AAA is developed.

Improving patient success with oral contraceptives: the importance of counseling.

Oral contraceptives (OCs) are highly effective in protecting against pregnancy if used correctly and consistently. They also offer noncontraceptive health benefits. The prescriber must understand the myriad factors contributing to success with OCs and other birth control methods, including characteristics and habits of successful users, real and perceived fears about adverse effects, and packaging issues. A woman's experiences with previous methods influence later success or failure with OCs.

PIP: Individualized counseling is an essential prerequisite to effective, consistent oral contraceptive (OC) use. Prescribers must understand the many factors that determine OC compliance, including characteristics and habits of individual users, a woman's experience with previous contraceptive methods, real and perceived fears about adverse effects such as menstrual irregularities, and packaging features that can impede or facilitate proper usage. A new OC user should be seen again in 2-3 months to assess her pill-taking habits. Continuing users can benefit from reminders to take the pill at the same time every day, to keep a supply of extra pills and back-up contraception, to use condoms with new partners, and to reread periodically the patient education handouts. Included with this article is a two-page sample patient education handout on OC instructions and information.