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Biomedical subjects

U Raju

Publications and source records attributed to U Raju.

At least 37 records · Page 2Linked to original sources

Breast-gut connection: origin of chenodeoxycholic acid in breast cyst fluid.

The notion that a breast-gut connection might modulate the microenvironment of breast tissue was supported by the finding that breast cyst fluid contains bile acids that are characteristically found in the intestines. To establish that the gut, rather than circulating steroid precursors, is the source of bile acids in breast cyst fluid, we gave two patients deuterium-labelled chenodeoxycholic acid (three 200 mg doses by mouth), starting 9 days before aspiration of breast cysts. The chenodeoxycholic acid concentration of seven samples of aspirated cyst fluid ranged from 42 to 94 mumol/L. The corresponding serum concentrations of chenodeoxycholic acid on the same day were 0.8 and 2.9 mumol/L, of which the labelled compound comprised 13.0% (0.38 mumol/L) and 28.2% (0.23 mumol/L). The deuterated chenodeoxycholic acid concentrations in cyst fluid were 0.79 and 1.26 mumol/L in two samples from patient 1 and 3.22 mumol/L in patient 2; these values are equivalent to 11-17% of the serum concentrations [corrected]. This study shows that intestinal bile acids rapidly gain access to cyst fluid. Further studies should investigate the mechanisms that govern the exchange processes and the maintenance of the high cyst fluid to plasma concentration gradients, and the biological half-lives of individual constituents.

Adult↗

Noninvasive breast cancer.

Noninvasive breast cancer comprises two distinct entities, lobular carcinoma in situ (LCIS) and ductal carcinoma in situ (DCIS). The increased use of screening mammography has caused more cases of noninvasive breast cancer to be discovered. However, the increased detection of LCIS and DCIS has posed problems in patient care for the radiologist, pathologist, surgeon, radiation oncologist, and medical oncologist. The authors review the past history of LCIS and DCIS, as well as the diagnostic challenges and therapeutic considerations for the multidisciplinary breast cancer team.

Breast↗

The histologic spectrum of apocrine breast proliferations: a comparative study of morphology and DNA content by image analysis.

Apocrine features occurring in sclerosing adenosis (apocrine adenosis), atypical ductal hyperplasia (ADH), and non-comedo ductal carcinoma in situ (DCIS) often add to diagnostic difficulty. We have evaluated the usefulness of DNA content determined by image analysis in apocrine metaplasia and hyperplasia, apocrine adenosis, ADH, DCIS, lobular carcinoma in situ, invasive ductal carcinoma, and infiltrating lobular carcinoma as a potential diagnostic aid in some of these problematic breast lesions with apocrine features. Infiltrating ductal carcinoma and DCIS were further subdivided into high- or low-grade category based on nuclear features. Microscopic fields containing 63 lesions were identified in slides from breast excisions. From each selected area 100 cells in corresponding fields in paired Feulgen-stained sections were digitized for computerized ploidy analysis with lymphocyte nuclei in the same slides serving as internal diploid controls. Aneuploidy was assessed using combined DNA index and modified Auer histogram criteria for DNA content abnormalities. There was strong association between the assessment of nuclear grade and ploidy (Fisher's exact test, P < .00001). All but one of the benign and low-grade malignant lesions (97%) were in the diploid range (six of seven apocrine metaplasia cases, three of three apocrine adenosis cases, 14 of 14 ADH cases, 10 of 10 low-grade DCIS cases, two of two lobular carcinoma in situ cases, and two of two infiltrating lobular carcinoma cases). In contrast, 24 of 25 (96%) of the high-grade malignant lesions were aneuploid (10 of 10 DCIS cases and 14 of 15 infiltrating ductal carcinoma cases). We conclude that DNA ploidy status does not offer additional diagnostic information to light microscopy in distinguishing among benign apocrine proliferations, ADH, and low-grade DCIS since these proliferations share a diploid range DNA content.

Breast Neoplasms↗

Identification of an annexin-like protein and its possible role in the Aplysia eye circadian system.

Light and serotonin regulate the phase of the circadian rhythm of the isolated eye of Aplysia. To screen for possible protein components of the eye circadian oscillator, we identified a number of proteins whose synthesis was altered in opposite ways by light and serotonin. The cellular function of one of these proteins was investigated by obtaining a partial amino acid sequence of it and by examining its immunoreactivity. A 38-amino acid sequence was obtained from a 40-kDa (isoelectric point 5.6) protein. A greater than 60% amino acid identity existed between this sequence and sequences of a family of calcium/phospholipid-binding proteins called annexins. Furthermore, the 40-kDa protein reacted with antibodies generated against a conserved amino acid sequence of annexins and with antibodies raised against human annexin I. The identification of the 40-kDa, light- and serotonin-regulated protein as an annexin led us to hypothesize that arachidonic acid metabolism plays a role in the Aplysia eye circadian system. To test this hypothesis, we examined the ability of an inhibitor of the arachidonic acid metabolic pathway to perturb the eye rhythm. Pulse treatments of isolated eyes with a lipoxygenase inhibitor, nordihydroguaiaretic acid, phase shifted the rhythm. The phase-shifting ability of nordihydroguaiaretic acid suggests that arachidonic acid and some of its metabolites may play a role in the eye circadian system. The results of our studies raise the possibility that links may exist between the 40-kDa annexin-like protein, arachidonic acid metabolism, and the circadian oscillator.

Acetophenones↗

Signet ring variant of lobular carcinoma of the breast: a clinicopathologic and immunohistochemical study.

Signet ring carcinoma of the breast often metastasizes to gastrointestinal tract and female genital tract. We report clinicopathologic features of 10 breast carcinomas with signet ring features, five of which had unusual metastatic patterns. The primary breast tumor in all these cases was lobular carcinoma. Although signet ring cells were prominent in metastatic sites, the primary tumor lacked signet ring cells in two cases. A linitis plastica-like presentation and presence of signet ring cells in gastric metastases raised a strong possibility of primary gastric carcinoma in three cases. The monoclonal antibody to gross cystic disease fluid protein (GCDFP-15) was positive in signet ring cell-rich areas in the primary breast tumor (8/10) and/or in the metastases in all cases. For comparison we studied GCDFP-15 immunoreactivity in 10 infiltrating lobular and 10 infiltrating ductal breast carcinomas with no obvious signet ring cells, and in 14 signet ring carcinomas from other sites (10 gastric, 2 prostatic, 2 colonic). The gastric, colonic, and one prostatic signet ring carcinoma were nonreactive. One prostatic signet ring carcinoma exhibited focal but unequivocal positivity with GCDFP-15. The cases of this report reinforce the concept that signet ring carcinoma of the breast is usually a variant of lobular carcinoma and not a distinct entity. Signet ring cell predominance in metastases, even in the absence of signet ring cells in the primary tumor, attest to the morpho-functional heterogeneity of lobular carcinoma. GCDFP-15 is a sensitive marker for signet ring breast carcinoma and a very useful adjunct tool in the diagnosis of metastatic signet ring carcinoma of mammary origin.

Adult↗

Esterase activity in human breast cyst fluid: associations with steroid sulfates and cations.

Human breast cyst fluid (BCF) contains an esterase that on the basis of electrophoretic mobility and response to inhibitors differs from those found in the plasma. From a total of 384 BCF samples analyzed for esterase using p-nitrophenyl hexanoate as substrate, 149 (39%) showed significant activity. The samples had been analyzed for the concentrations of the sulfates of estrone, estriol, dehydroepiandrosterone, as well as the potassium and sodium cations (K+/Na+). The data were submitted to statistical analysis using the Spearman rank order test. The esterase-positive samples exhibited a significant positive association with each of the steroid sulfates and the K+/Na+ ratios. Except for protein concentration, there was no significant correlation between the esterase-positive and esterase-negative cysts. These observations may have physiological significance in that high K+/Na+ ratio cysts have been related to the histological status of the cyst.

Biomarkers↗

Relationship between the concentrations of estriol sulfate and estrone sulfate in human breast cyst fluid.

Estriol-3-sulfate (E3S) is present in human breast cyst fluid (BCF) in median levels of 8.7-10.4 nmol/L, yet is barely detectable in the serum (less than 0.034 nmol/L). The source of this huge concentration of E3S is unknown. It may accumulate from blood by active transport or be synthesized and concentrated within the cyst. Since estrone sulfate (E1S) and its possible precursor, dehydroepiandrosterone sulfate (DHEAS) are elevated in BCF, E3S may originate via 16 alpha-hydroxylation of E1S. The present study examined the correlations between the levels of DHEAS and E1S with those of E3S in BCF. The sodium and potassium ions were also quantified and related to the steroid concentrations. By linear regression analysis of log-normalized data there was a highly significant correlation between the concentrations of E1S and E3S (n = 355, r = 0.690, P less than 0.001) and between DHEAS and E3S (n = 361, r = 0.577, P less than 0.001). The BCF were classified according to their K/Na ion ratios: type 1, greater than 1.0, type II, less than 0.25, and type III, 0.25-1.0. By Student's t test, the concentrations of E3S differed between each BCF Type (P less than 0.002). This was also true for E1S and DHEAS. Type 1 cysts were associated with the highest estrogen sulfate levels and type II with the lowest levels. The possible physiological importance of this observation resides in reports that the BCF type expressing the highest steroid concentrations has been related to an aporcine-like epithelial lining of the cyst wall and a somewhat higher risk for developing breast cancer. The results suggest that E3S in BCF may originate from E1S, but alternate mechanisms are not precluded.

Aryl Hydrocarbon Hydroxylases↗

Steroid and cation correlations in human breast cyst fluid: preliminary findings.

Gross cystic disease (GCD) of the breast imparts a minimal risk for the development of breast cancer. Relative to serum, breast cyst fluid (BCF) in patients with GCD is highly concentrated in androgens, estrogens, certain enzymes, and bioactive polypeptides. In addition, the cations, sodium and potassium, vary markedly and inversely in BCF. We have focused on the levels of estriol-3-sulfate (E3S), which are several orders of magnitude greater in BCF than in blood. In this preliminary study, about 55 specimens of BCF were analyzed for E3S, its possible precursors, estrone sulfate (E1S) and dehydroisoandrosterone sulfate (DHAS), and the cations, sodium and potassium. The data were analyzed statistically by linear regression analysis. E3S correlated directly with both E1S and DHAS (p less than 0.002), and inversely with the Na/K ratio (p less than 0.003). Low Na/K ratio has been associated with secretory processes in cyst epithelium. The data suggest that E3S may originate via 16 alpha-hydroxylation of estrogen in the cyst and that elevated E3S levels may be indicative of a secretory epithelium. This is part of an ongoing prospective study involving 400 subjects with GCD to see whether hormonal and enzymic profiles can be related to cancer risk.

Aryl Hydrocarbon Hydroxylases↗

Alteration of the phase and period of a circadian oscillator by a reversible transcription inhibitor.

A function for transcription in the mechanism of a circadian oscillator was investigated with the reversible transcription inhibitor 5,6-dichloro-1-beta-D- ribobenzimidazole (DRB). Two-hour treatments with DRB shifted the phase of the circadian rhythm of the isolated eye of Aplysia, and continuous treatments of DRB lengthened the free running period of this rhythm. Camptothecin, an inhibitor of transcription that is structurally unrelated to DRB, had similar effects on the circadian rhythm. These results suggest that transcription may be part of the circadian oscillating mechanism.

Animals↗

A comparative immunohistochemical study of peritoneal and ovarian serous tumors, and mesotheliomas.

The distinction between serous neoplasms of the peritoneum in women and conventional mesothelioma can be difficult. In order to determine any significant immunohistochemical differences, formalin-fixed, paraffin-embedded sections of 10 peritoneal serous tumors (PST), 10 ovarian serous tumors (OST), and 10 epithelial mesotheliomas were evaluated with a panel of 10 antibodies directed against carcinoembryonic antigen (CEA: polyclonal, monoclonal), high molecular weight keratin (34 beta E12), low molecular weight keratin (35 beta H11), Leu-M1, TAG-72 (monoclonal antibody B72.3), human milk fat globulin (HMFG-2), vimentin, placental alkaline phosphatase (PLAP), and S-100 protein. The antibodies CEA, Leu-M1, and B72.3 had the most discriminatory value in differentiating serous tumors from mesothelioma. Eighty-five percent of PSTs and OSTs (17 of 20) were positive with CEA, Leu-M1, and/or B72.3. None of the mesotheliomas stained for CEA or Leu-M1; three mesotheliomas had very focal positivity with B72.3 (1% or less). Vimentin, PLAP, HMGF-2, keratin, and S-100 had no significant discriminatory value. Epithelial mucin was present in 80% of serous tumors, while the mesotheliomas lacked epithelial mucin. Leu-M1, CEA, and/or B72.3 positivity in a peritoneal tumor supports a diagnosis of serous tumor. However, since some PST do not stain for any of the three antibodies and the focal nature of positive reactions in some cases may be difficult to interpret, exclusion of mesotheliomas is enhanced by the use of mucin stains.

Adult↗

Epitheliosis of the breast. An immunohistochemical characterization and comparison to malignant intraductal proliferations of the breast.

Epitheliosis is a benign intraluminal proliferation in the breast ducts and lobules that needs to be distinguished from ductal carcinoma in situ (DCIS). The histogenesis and differentiation of cells comprising epitheliosis have been the subject of some controversy. We evaluated the expression of a high-molecular-weight keratin (34 beta E12), muscle-specific actin (HHF-35), and S-100 protein immunoreactivity in formalin-fixed sections of the breast with epitheliosis and DCIS. In 28 of 30 cases of epitheliosis, there was strong HMW keratin immunoreactivity in the streaming sheetlike intraluminal proliferations. In contrast, 35 of 40 cases of DCIS (nonpapillary and papillary) were nonreactive for HMW keratin; the other five were weakly reactive. Furthermore, in 10 cases of DCIS, some ducts had isolated or small aggregates of HMW keratin-positive benign cells on the luminal aspects of the neoplastic proliferation that were reminiscent of a pagetoid pattern. Muscle actin-stained sections were analyzed to assess myoepithelial (ME) cell participation in epitheliosis. Muscle actin-positive ME cells were present at the periphery of the involved ducts but were absent or rare within epitheliosis. The distribution of ME cells--i.e., at the periphery of the spaces involved--was similar in DCIS and epitheliosis. S-100 protein was weakly but relatively consistently expressed by epitheliosis, but all cases of DCIS were negative. Six cases of atypical ductal hyperplasia included in the study were negative for HMW keratin, muscle actin, and S-100 protein. The immunohistochemical profile of epitheliosis indicates that it is primarily an epithelial proliferation with strong HMW keratin and weak S-100 protein expression but without ME cell participation. The distinct differences in HMW keratin expression of epitheliosis and intraductal carcinoma appear to reflect a consistent antigenic difference in these two biologically distinct forms of proliferation.

Actins↗

Involvement of proteins in light resetting ocular circadian oscillators of Aplysia.

The effect of inhibitors of protein synthesis on the phase shifting action of light was investigated. Anisomycin and cycloheximide appeared to block advance phase shifts produced by light. This result suggested that light might phase shift by changing the synthesis of some proteins. Examining proteins separated by two-dimensional gel electrophoresis, we found that incorporation of amino acids into 11 proteins was changed during a 6-h light pulse. Nine of these 11 proteins were affected by light in a phase-dependent manner. Elevated extracellular potassium and 8-bromo-guanosine 3',5'-cyclic monophosphate (cGMP), two treatments that mimic effects of light on the rhythm, also changed amino acid incorporation into a number of proteins. All of the five proteins affected by 8-bromo-cGMP were also affected in the same manner by light. Three proteins were affected similarly by elevated potassium, light, and 8-bromo-cGMP. Exposure of eyes to label at different times after light treatment showed that the effects of light on some proteins were long lasting. In addition, some proteins were not affected during light but were affected only several hours after light. Some of the eye proteins affected by light were also altered by serotonin (5-HT), another phase-shifting agent. The proteins affected by light, elevated potassium, 8-bromo-cGMP, and 5-HT are candidates for components of the circadian system either as an element of the entrainment pathway or the oscillator mechanism.

Amino Acids↗

Bile acids in human breast cyst fluid: the identification of lithocholic acid.

Breast cyst fluid (BCF) aspirated from 12 women with fibrocystic disease of the breast and sera obtained simultaneously were analyzed for bile acids. Analysis was performed by gas-liquid chromatography of the acetoxy methyl esters of the bile acids prepared after alkaline hydrolysis of the bile salts. An internal standard served to correct for methodological losses. Low levels of bile acids were found in serum samples, precluding overt hepatobiliary complications. Deoxycholic acid (17-160 mumol/L), chenodeoxycholic acid (18-305 mumol/L), and cholic acid (3-119 mumol/L) were detected in 11 of 12 samples of BCF. In 2 cases, chosen at random, the identities of the bile acids were verified by mass spectrometry. Lithocholic acid (9-23 mumol/L), a reported cocarcinogen, was detected in 6 of the 12 samples of BCF. This is the first report of the presence of lithocholic acid in BCF with confirmation by Mass spectrometry. There was no correlation between the levels of individual bile acids and those of potassium ion, Na+/K+, estriol-3-sulfate, or 16 alpha-hydroxyandrogen sulfates that had been quantified previously in these samples. There was borderline correlation between concentrations of total bile acids and K+ (P less than 0.06) and Na+/K+ (P less than 0.07). Yet to be elucidated are the mechanism of accumulation of bile acids in BCF and whether levels of particular bile acids in BCF may serve to identify that small subset of women with fibrocystic disease at risk for developing breast cancer.

Adult↗

Myoepithelial cells in the differential diagnosis of complex benign and malignant breast lesions: an immunohistochemical study.

The distinction between sclerosing adenosis, radial scars, noninvasive carcinomas occurring in sclerosing adenosis, and invasive carcinoma can be difficult. The identification of a myoepithelial (ME) cell layer is helpful in establishing a diagnosis of complex benign breast proliferation as well as intraepithelial neoplasia in sclerosing adenosis. We reviewed pathologic material from patients with tubular carcinoma (23) and complex breast proliferations (28), including sclerosing adenosis (12), radial scars (9), sclerosing adenosis with intraepithelial neoplasia (5), and sclerosing adenosis with atypical apocrine metaplasia (2). Immunoperoxidase stains on formalin-fixed, paraffin-embedded tissue using a muscle actin-specific antibody of clone HHF35 and high molecular weight cytokeratin of clone 34 beta E12 (HMW keratin) were performed to identify myoepithelial cells. Muscle actin was uniformly reliable in staining ME cells, as well as other actin-containing cells such as myofibroblasts and vascular smooth muscle. HMW keratin was less reliable, being poorly sensitive and less specific than muscle actin for labeling of ME cells. ME cells were readily identified at the periphery of ductules in all complex benign breast lesions. The presence of ME cells distinguished intraepithelial neoplasia involving sclerosing adenosis from invasive carcinomas. Well differentiated invasive carcinoma forming tubular structures lacked a ME cell layer.

Actins↗

ECG changes in asphyxia neonatorum.

Twenty five asphyxiated neonates had ECG changes consistent with degree of asphyxia. Equivocal changes were found in mild asphyxia and changes suggestive of myocardial infarction were seen with severe asphyxia. In most cases, the changes reverted to normal within two weeks signifying great ability of the neonatal heart to withstand hypoxic insult. Four babies with severe asphyxia having ECG changes suggestive of acute myocardial infarction expired within 48 hours of birth.

Asphyxia Neonatorum↗

The concentration of 16x-hydroxy androgens in serum and cyst fluid of women with gross cystic disease of the breast.

The concentrations of 16 alpha-hydroxydehydroepi-androsterone sulfate (16 alpha-OHDHAS) and androst-5-ene-3 beta, 16 alpha-, 17 beta-triol-3-sulfate (A-TriolS) were measured in the plasma and breast cyst fluid (BCF) of women with gross cystic disease of the breast. In the 19 BCF samples analyzed, the 16 alpha-OHDHAS and A-TriolS concentrations ranged from 15 to 1130 ng/mL, and 12 to 871 ng/mL, respectively. However the concentrations of these steroids in the sera of these women were lower (15-179 ng/mL, 8-80 ng/mL, respectively). Estriol-3-sulfate (E3-3S) concentrations in the BCF samples ranged from barely detectable (0.2 ng/mL) to 3 micrograms/mL. In BCF or serum a positive linear correlation was observed in the concentration of 16 alpha-OHDHAS and A-TriolS (p less than 0.001 and 0.05, respectively). However, in the same patients no statistical significance was observed in the BCF vs serum concentrations of these two steroids. When the specimens from this and previous studies were combined, positive correlation was found between potassium ion concentration and E3-3S or 16 alpha-OHDHAS. The origin of the high concentration of E3-3S is still obscure. Although no linear correlation between 16 alpha-OHDHAS and E3-3S was observed, the possibility of a precursor-product relationship between the two is not elimnated.

Androgens↗

Primary papillary serous neoplasia of the peritoneum: a clinicopathologic and ultrastructural study of eight cases.

The well-documented but rare primary papillary serous peritoneal tumors are difficult problems for the pathologist and the clinician. Because of their unusual location, these tumors are often classified as mesothelioma or advanced ovarian carcinoma. In this study, we report the clinicopathologic features of eight primary peritoneal serous papillary tumors and compare their histologic and ultrastructural features to 12 serous ovarian tumors and 16 epithelial mesotheliomas (two peritoneal and 14 pleural). The eight peritoneal serous papillary tumors occurred in women aged 19 to 75 years; two were serous tumors of low malignant potential (borderline) and six were serous carcinomas. The tumors were located in the mesosalpinx, left pelvis, omentum, and/or surface of the ovary. The two patients with borderline neoplasms had long disease-free survival (11 years and 20 years), while three of the four patients with carcinoma with more than 1 year of follow-up died of disease. The peritoneal serous papillary tumors were morphologically identical to serous ovarian tumors of equivalent grade. Well-differentiated papillary structures with distinct fibrovascular cores and one or several layers of columnar, crowded cells, dense overlapping nuclei with a long axis perpendicular to the surface of the papillary cores, and numerous psammoma bodies were features of the peritoneal and ovarian serous tumors. In contrast, the tubulo-alveolar, solid, or poorly defined papillary structures lined by well-spaced polygonal to cuboidal cells with abundant cytoplasm, absence of nuclear polarity, and infrequent psammoma bodies characterized the mesotheliomas. Epithelial mucin and carcinoembryonic antigen (CEA) immunoreactivity, when present, supported a diagnosis of serous tumor in these generally mucin-poor and CEA-negative neoplasms. Ultrastructurally, the cells of serous tumors had slender, straight microvilli of variable length interspersed with or without cilia, while the nonciliated cells of mesothelioma had long, exuberant, wavy microvilli. The morphologic and clinical features of the peritoneal papillary serous tumors are distinctive enough to warrant their separation from mesotheliomas.

Adult↗