Retinoic acid-binding protein in human breast cancer and dysplasia.

Seventy-five specimens of human breast tissue were checked for the presence of cellular retinoic acid-binding protein (cRABP). Fifty-two percent of the primary carcinomas and 43% of the dysplastic breast lesions (stage MII) contained detectable amounts of crabp, whereas no cRABP was found in normal tissue. Sucrose gradient centrifugation and electrophoresis on agarose were used for analysis of the presence of cRABP. The cRABP of human origin (normal uterus and neoplastic mammary tissue) differed in its mobility in agarose electrophoresis from that of rat testis cRABP.

Quantitative estimation of cellular retinoic acid-binding protein activity in normal, dysplastic, and neoplastic human breast tissue.

A technique for reproduction and quantitative determination of human cellular retinoic acid-binding protein (CRABP) activity in breast tissue specimens is described. A multiphasic polyacrylamide disc gel electrophoresis system (operative at pH 10.2) was adapted for this purpose. This technique allows, after incubation with tritiated retinoic acid (RA) overnight, the separation of the specific CRABP activity from the nonspecific serum-originated binding activity and from the free RA. Previous purification of the tissue cytosols is therefore not necessary. The same assay method was also used for the determination of the molecular weight (Ferguson plot, m.w. 13,000) and the dissociation constant Kd (2.5 x 10(-7) M) of mammary CRABP. The activity in tissue cytosol, stored at -70 degrees, was found to be stable for at least 3 months. Results from 88 breast tissue specimens of different pathological degree are presented. CRABP activity was found in all tissue categories with progressively increasing amounts from normal tissue to breast cancer. The activity in the cancer tissues (14.85 +/- 12.05 pmol RA bound per mg soluble protein: N = 27) was significantly different (p less than 0.001) from the activity determined in tissue with simple dysplasia without epithelial proliferation [4.3 +/- 2.2 (S.D.) pmol RA bound per mg protein; N = 30]. It is possible that in the cases where high amounts of CRABP activity are found in dysplastic and preneoplastic tissue, a high risk for breast cancer development exists. Therefore, CRABP is tentatively proposed as a dedifferentiation and/or proliferation marker.

Interstitial hyperthermia and high dose rate brachytherapy in the treatment of anal cancer: a phase I/II study.

PURPOSE: The rate of local failure is sufficiently high following sphincter conserving surgery and radiation therapy for advanced anal cancers to warrant investigation of improved local treatment techniques. This Phase I/II study was undertaken to investigate the site-specific toxicities and response of Stage II and III anal cancers to interstitial thermoradiotherapy using a hot water interstitial system. METHODS AND MATERIALS: Between September 1988 and March 1991, 14 patients with primary carcinomas of the anal canal, UICC Stage T2-3, N0-1, M0, were treated with split-course external beam irradiation to the pelvis (30 Gy + 20 Gy) and 1 or 2 interstitial Iridium-192 high dose rate (Ir-192 HDR) implants (6-8 Gy each) immediately followed by interstitial hyperthermia (HT). Patients with tumor diameters > 3 cm were scheduled to receive chemotherapy consisting of 2 courses of 5-fluorouracil and mitomycin C given concomitantly with external beam radiation. Interstitial hyperthermia was induced by circulating warm water through the needles that were implanted to hold the Ir-192 source. The treatment goal was to achieve and maintain a temperature of 42.5 degrees C over a time period of 40 min. A 3-point thermocouple probe inserted into one or two additional needles was used for thermometry. The temperatures were recorded by manual mapping along these needles at steps of 0.5 or 1 cm. RESULTS: A total of 20 Ir-192 HDR-HT implants were performed in 14 patients. All but two patients completed the external beam irradiation; five patients received concomitant chemotherapy. Analysis of thermal parameters showed that minimum intratumoral temperatures (Tmin) of 42 degrees C, 42.5 degrees C, 43 degrees C, and 44 degrees C were achieved in 64%, 37.5%, 14%, and 7% of patients, respectively. Intratumoral mean Tmin, mean average, and mean maximum temperatures for these patients were 41.7 degrees C, 42.4 degrees C, and 43.4 degrees C, respectively. Brachytherapy and HT were well tolerated. Clinical complete responses (cCR) were obtained in 11/14 (78.5%) patients, complete histopathological responses (pCR) in 10/14 (71%). Only one patient with pCR recurred and succumbed to her disease. Patients with persistent disease (1 minimal and 3 partial responders, including 1 cCR) underwent abdominal-perineal resection but subsequently died from local-regional recurrence. One patient with pCR died from unrelated causes. Median survival for all patients from onset of radiation to death or last follow-up is 26 months. Eight patients are alive disease-free after a follow-up ranging from 16-44 months (median: 30, mean: 30 months). Treatment complications were limited to two patients who developed persistent ulcers. Sphincter function was maintained in 50% of patients. CONCLUSION: This study demonstrates that interstitial warm water hyperthermia in combination with brachytherapy for anal carcinomas is feasible and did not add to complications when compared to studies employing external beam irradiation and brachytherapy alone. The thermal parameters obtained by the warm water system compare favorably to those reported by others using radiofrequency and microwave systems.

Experience with split-course external beam irradiation +/- chemotherapy and integrated Ir-192 high-dose-rate brachytherapy in the treatment of primary carcinomas of the anal canal.

PURPOSE: The effect of the treatment of anal cancer by performing a high-dose-rate (HDR) brachytherapy boost during a short split between the external beam radiotherapy series (EBR) +/- chemotherapy was investigated. METHODS AND MATERIALS: Thirty-nine patients with anal canal cancers, stages T1-T4 N0-2 M0, were treated with split-course EBR (50-50.4 Gy) and a Iridium 192 ((192)Ir-) HDR boost (6 Gy) performed during the 1-2-week split. Patients who failed to achieve a complete tumor response received additional brachytherapy. Chemotherapy with 5-fluorouracil and mitomycin C was offered to patients with tumors > 3 cm and employed concomitantly on days 1-5 and day 1, respectively, of each EBR series. RESULTS: Follow-up ranged from 3 to 140 months (median 31). Median treatment duration was 56 days. The 3-year (5-year) actuarial rates of locoregional control (LRC) and disease-specific survival (DSS) were 81% (76%) and 80% (76%), respectively. The crude rate of anal preservation was 77% overall, and 97% in patients in whom LRC was achieved. Uncompromised anal function was recorded in 93% of these patients. The actuarial 3-year (5-year) rate of colostomy-free survival (CFS) was 78% (73%). There was a statistically significant difference in LRC and DSS according to stage, tumor size, and nodal status. Complications requiring surgical intervention occurred in 7.6% of patients. CONCLUSION: The integration of the HDR boost in a split-course EBR regimen +/- chemotherapy resulted in excellent sphincter function without an increase of severe complications and with rates of LRC, DSS, and CFS, which compare favorably with those reported in the literature.

Comparative serological reactivity of Taenia crassiceps, Taenia solium and Taenia saginata metacestode neutral glycolipids to infection serum from Taenia crassiceps-infected mice.

A comparative survey was undertaken of the neutral fraction glycolipids from the metacestodes of 3 taeniid species, Taenia crassiceps, Taenia solium and Taenia saginata, to determine their chemical and serological staining patterns on separation by thin-layer chromatography. The orcinol-positive patterns of T. solium and T. saginata metacestodes exhibited a closer superficial resemblance to each other than to T. crassiceps or T. saginata adults. A comparison of component migration properties against standards of known structure indicated the main oligosaccharide chains to be mono-, di-, tri- and tetrasaccharides; however, in T. solium this was extended to at least a heptasaccharide. The multiple banding characteristic of each component is a consequence of lipid moiety heterogeneity. Serologically, the patterns of the 3 taeniid species neutral fraction glycolipids showed virtually the same immunological reactivity towards mouse normal serum, infection serum and a monospecific, polyclonal antibody directed against the trisaccharide component of T. crassiceps. The latter antibody was isolated from mouse infection serum by affinity chromatography on a column of glycolipid-bound octyl-Sepharose CL-4B. Immunochemically, the major common epitope expressed by the neutral fraction glycolipids of the 3 taeniid species is the same or very similar to the glycosphingolipid, neogalatriaosyl ceramide derived from the marine mollusc Turbo cornutus (Gal(beta 1-6) Gal(beta 1-6) Gal(beta 1-1)Cer). Host tissue neutral fraction glycolipids, porcine muscle and bovine muscle, as well as human spleen, were not immunoreactive.

Intraoperative radiation therapy in advanced thyroid cancer.

An intraoperative radiation therapy (IORT)-protocol was designed for poorly differentiated non-anaplastic thyroid carcinoma. Out of 155 cases of differentiated thyroid tumours, 12 showed marked vascular/capsular invasion. Five entered the study (three primarily local invasive tumours, two local recurring). IORT was administered after tumour surgery (4-10 GY) and combined with post-operative percutaneous irradiation. The tumour control rate in the thyroid bed was achieved in all five patients, 1/5 developed mediastinal nodes and 1/5 with primary mediastinal tumour extension showed tumour progression. No specific complications occurred.

Endocavitary Ir-192 radiation and laser treatment for palliation of obstructive rectal cancer.

Endoscopic laser therapy (ELT) either alone or combined with endocavitary Ir-192 radiation is performed for advanced, inoperable rectal cancer and when patients are ineligible for surgery due to severe concomitant medical illness. During the period from January 1984 to January 1997 we treated 81 patients (51 males, 30 females). Sixty-seven patients had ELT only using a ND-Yag Laser system. Twenty-five patients (average age: 80.5 years) were ineligible for surgery (Group I). Forty-two patients (74.1 years) had an advanced locally inoperable tumour (Group II). Fourteen patients (76.5 years) underwent a combined therapeutic regime with endocavitary Ir-192 afterloading following ELT (Group III). Adequate desobliteration was achieved in 100% (groups I and III) and 97% (group II) of the patients. The average interval to aftertreatment was 8.4 weeks in group I and 9.4 weeks in group II, compared to 11.5 weeks in group III. Serious complications (perianal abscess, rectovaginal fistula) occurred in 3.7%, minor complications (laser-induced bleedings, unclear fever) in 12.3%. All laser-induced bleedings could be dealt with using laser therapy. The frequency of treatment was governed by tumour mass and the patient's survival. The results suggest that additional endocavitary radiation significantly prolongs the maintenance of normal bowel function compared with laser therapy alone.

Evaluating intraoperative radiation therapy (IORT) and external beam radiation therapy (EBRT) in non-small cell lung cancer (NSCLC). Five years experience.

A pilot study on intraoperative radiation therapy (IORT) combined with external beam radiation therapy (EBRT) in nonresectable non-small cell lung cancer (NSCLC) was performed in 31 patients (mean age: 66.2 years, range: 51-80; 10 anatomically and functionally, 21 functionally, nonresectable; 20 squamous-cell, 11 adenocarcinoma). The tumor was exposed by lateral thoracotomy and a staging lymph node dissection was performed (final staging 7 T1, 16 T2, 8 T3; 11 nodal positive). Ten to 20 Gy IORT (energy: 7-20 MeV electrons) were delivered to the tumor. Unilateral continuous positive airway pressure ventilation of the diseased lung was used to reduce the amount of healthy lung tissue in the IORT port and to minimize the ventilatory movement. Secondary collimation and direct shielding of radio-sensitive structures within the IORT port by aluminium sheets were used to further reduce collateral damage. Four weeks after IORT, 46 Gy EBRT (2 Gy/day 5 times a week; 8-23 MeV photons) were administered to the mediastinum and to the tumor-bearing area on an outpatient basis. In nodal positive cases the mediastinal dose was increased to 56 Gy. Twenty-three patients were evaluable. In 13 complete, in 8 partial (50-97% regression) and in 2 minor response has been achieved. Five patients experienced a recurrence (local only: 2; local and distant: 1; distant only: 2). Twelve patients died of underlying cardio-respiratory disorders within 6 to 25 months after IORT; 7 died of cancer. The overall 5-year survival rate including the incidental deaths is 14.7%. The recurrence-free survival rate is 53.2%.

Inhibition of in vitro and in vivo mast cell degranulation by Taenia crassiceps metacestodes in vitro incubation products.

In vitro released products of T. crassiceps metacestodes (TcIP) harvested from the peritoneal cavity of NMRI mice were tested for inhibitory effects on the in vitro degranulation of peritoneal mast cells (MCs) of normal mice (NMRI) and rats (Wistar) and on the in vivo degranulation of rat (Wistar) skin MCs (PCA-assay). In vitro degranulation was elicited chemically (compound 48/80, polymyxin B or the bee venom peptide, mellitin). In vivo degranulation was triggered immunologically (anaphylactic systems ovalbumin/anti-ovalbumin or Fasciola hepatica crude fluke extract antigen/serum of fluke-infected rats (Wistar]. In vitro degranulation of murine peritoneal MCs or the in vitro histamine release of rat peritoneal MCs normally induced chemically was significantly inhibited when the MCs were preincubated with the TcIP or with serum of T. crassiceps-infected NMRI mice from day 35 post infection and thereafter. In vitro degranulation of peritoneal MCs of infected mice was strongly inhibited beginning on day 10 after infection. Also in vivo degranulation of the IgE-sensitized rat skin MCs was significantly reduced by intradermal injection of the TcIP before (6, 3 and 1 h) antigen challenge and by preinjection (1 h) of serum from infected mice (day 80 p.i.)-The inhibitory effect was also demonstrated after immunoadsorption of mouse serum proteins naturally contaminating the TcIP. Heating (100 degrees C/15 min), even in the presence of 0.25 M HCl, did not suppress the inhibitory activity.

Experimental dicrocoeliasis: the humoral immune response of golden hamsters and rabbits to primary infection with Dicrocoelium dendriticum.

The results presented deal with the humoral immune response of golden hamsters to primary experimental infection with D. dendriticum. The development of serum antibodies has been comparatively investigated with three hamster groups (n = 43) harbouring different burdens of adult flukes. The mean numbers of parasites were 11, 30, or 130 per animal. Serum antibody response was studied during an observation period of at least 331 and up to 496 days postinfection. For antibody detection the sensitivities of precipitation test (PTs) (double diffusion test, immuno- and counterimmunoelectrophoresis), of the indirect haemagglutination test (IHAT), the complement fixation test (CFT), and the enzyme linked immuno sorbent assay (ELISA) were compared using aqueous crude fluke antigen and crude egg antigen. CFT and ELISA were most sensitive for the early detection of initial response. Thereafter all the tests employed revealed increasing antibody titres, which in general remained at constant levels and persisted until the end of the observation period with the exception of CF-antibodies. In general fluke antigen was found to be more sensitive than egg antigen. However, in CFT this antigen occasionally has been associated with unspecific inhibition of haemolysis. Comparison of the results shows that ELISA using crude fluke antigen gave the most realistic picture of the actual fluke burden. Also preliminary results on the precipitin response of rabbits (n = 3) after primary experimental exposure to different numbers of metacercariae (500, 1,000, and 3,000 per animal respectively) are reported. Employing the above mentioned PTs a persisting antibody response could be demonstrated only after exposure to at least 3,000 infective larvae. The initial response was found on day 63, the observation period was 550 days.

Purification of an antigen from the Taenia crassiceps metacestode vesicular fluid highly sensitive in detecting IgG-antibodies (ELISA) in calves with experimental cysticercosis.

This study reports on the purification of an antigen predominant within the vesicular fluid (VF) of T. crassiceps metacestodes and shown to share identity with the major vesicular fluid protein of the T. saginata larval stage. Purification was achieved by gel filtration of the VF on Sephacryl S-300 superfine, followed by ion exchange HPLC. The antigen represents a single polypeptide chain (Mr appr. 37,000) with carbohydrate moieties without affinity to ConA. Isoelectric focusing of the electrophoretical and immunological homogeneous antigen resulted in five bands focusing at pH 4.0, 4.2, 4.4, 4.6 and 4.8, respectively. In ELISA, the purified antigen detected serum IgG-antibodies in all 21-35 weeks old calves (n = 10) with experimental cysticercosis (70 to 6,000 viable larvae recovered). When compared to T. saginata metacestode VF the antigen was the better reagent for discriminating between infected and non-infected animals. As shown by immunodiffusion and ELISA the antigen is also common to the T. saginata adult stage and obviously to other taeniid metacestodes where it is accumulated in the VF or hydatid fluid.

Comparison of the in vitro translation capacity of Taenia crassiceps metacestode mRNA prepared by the phenol and cesium chloride method.

Total RNA of T. crassiceps metacestodes harvested from male and female NMRI mice was prepared by both the phenol extraction technique and cesium chloride (CsCl) gradient centrifugation. mRNA was selected by oligo (dT)-cellulose affinity chromatography and used as the template for the in vitro translation of parasite polypeptides in a cell-free rabbit reticulocyte lysate. The template activity of the mRNA obtained after CsCl preparation was clearly higher, as shown by the amount of 35S-methionine incorporated into the translation products and by fluorographed SDS-PAGE of the synthesized labelled polypeptides. SDS-PAGE fluorographs of antigens encoded by the mRNA prepared by CsCl centrifugation and selected by immunoprecipitation using purified IgG antibodies of T. crassiceps-infected mice (day 80 postinfection) exhibited seven labelled polypeptides of about 65, 46, 45, 42, 34, 29 kDa and a predominant 20-kDa antigen. The latter polypeptide was the only one recognized by the antibodies amongst the in vitro translation products directed by mRNA prepared by the phenol method.

Retrospective conversion: a survey of UNYOC/MLA members.

The Upstate New York and Ontario MLA surveyed its members to determine which libraries are conducting retrospective conversion projects and their reasons and methods. All academic and medical school libraries that responded are converting, but only 16 of 39 hospital libraries and 14 of 23 other libraries have such plans. The majority of respondents intend to do the work themselves instead of contracting with a vendor. They highly recommend that a written plan be drawn up before proceeding with retrospective conversion.

Immunoelectrophoretic analyses of antigens shared by the vesicular fluid and cyst wall of Taenia crassiceps and Taenia saginata metacestodes.

In this study, the antigenic mosaic of the vesicular fluid (VF) and hydrosoluble cyst-wall extract (CWE) of T. crassiceps (Tc; harvested from mice) and T. saginata (Ts) metacestodes was analyzed by combined precipitation maps developed in single and bidimensional immunoelectrophoresis against the respective rabbit antiserum. Host-serum proteins demonstrated by immunodiffusion within TcVF (albumin, transferrin, IgG, and another five noncharacterized proteins), TcCWE (albumin, IgG, and six additional unknown proteins), TsVF (albumin) and TsCWE (albumin and IgG) were removed by immunoaffinity chromatography prior to immunoelectrophoretic analysis. Neither TcVF nor TcCWE contained demonstrable amounts of mouse IgM and IgA. In TcVF a total of 18 and in TcCWE a total of 36 parasitic antigens were recognized by the corresponding antiserum. In the case of TsVF and TsCWE, antiserum to the crude extract of T. saginata larvae developed a total of 26 and 30 precipitates, respectively. Examination of the precipitation maps developed by the respective heterologous antiserum (vice-versa testing) showed that both TcVF and TsVF contained ten antigens sharing identity. For TcCWE and TsCWE, nearly the same number of shared antigens (20 and 18, respectively) could be demonstrated. For screening of IgG antibodies against T. saginata metacestodes from heavily and moderately infected calves (n = 6) by ELISA, VF and CWE antigens of both Taenia species were found to be potent reagents; TsVF was the most sensitive antigen.

Distortion of grating prisms, and their use in radial velocity determinations of astronomical objects with slitless field spectrographs.

The basic theory of off-axis distortion and dispersion errors of grating-prisms is given. it is used to determine steller radial velocities according to the reversion method with slitless grating-prism field spectrographs.

Serodiagnosis of human onchocerciasis: evaluation of sensitivity and specificity of a purified Litomosoides carinii adult worm antigen.

Sensitivity and specificity of a Litomosoides carinii macrofilariae aqueous crude extract and a purified antigen which was isolated by preparative flat bed electrofocusing, were evaluated for the immunodiagnosis of human onchocerciasis in 3 serological tests: Double diffusion test (DT), latex agglutination test (LAT) and ELISA. Testing sera from proven cases of onchocerciasis (n = 28), loiasis (n = 4) filariasis bancrofti (n = 4), and other non filarial helminth infections (n = 29) in all tests the purified antigen was clearly more sensitive and specific. Testing onchocerciasis sera highest sensitivity was found by ELISA (92.9%) followed by latex agglutination test (89.3%) and double diffusion test (82.1%). Specificity controls of the purified antigen with sera from patients with helminth infections others than filariases led to 0% (DT), 6.9% (LAT) and 10.3% (ELISA) false positive reactions. When the 3 tests were used in combination in a way that at least two positive reactions in different tests were regarded as necessary to declare a serum as positive, sensitivity increased to 100% and unspecific reactions were found only testing one serum (3.4%) originating from a case of cystic echinococcosis.

[Immunization against Capillaria hepatica: the effects of primary infections, x-irradiated stages, non-embryonated eggs, and soluble egg extracts (author's transl)]. / Immunisierung gegen Capillaria hepatica: Die Wirkung von Erstinfektionen, röntgenattenuierten Stadien, nicht embryonierten Eiern und löslichen Eiextrakten.

The influence of primary infections with embryonated infective eggs or with X-irradiated infective eggs, and of non-embryonated eggs, and egg homogenate extracts on challenge infections with Capillaria hepatica was investigated. The worm reproductivity was significantly suppressed in a sublethal challenge infection given 11 days after a primary infection of Mastomys natalensis with 50, 150, 400, and 800 eggs per animal. The administration of 600 X-irradiated (2.2 Krd) embryonated eggs 36 days before challenge as well as an intraperitoneal injection of non-embryonated eggs 12, 10, 8, 6, 4, and 2 days before challenge (simulating the egg production of a normal infection) also reduced significantly the egg production of a weak (50 eggs/ animal) infection. No effect was observed on a moderate challenge (300 eggs/animal). The effect was not markedly enhanced by the repeated administration of X-irradiated eggs or by the combination of X-irradiated infective eggs and non-embryonated eggs. Immunization of mice with soluble egg extracts resulted in significant reduction of egg production determined 60 days after challenge. Two hundred and thirty eggs of C. hepatica/g body weight proved to be a lethal infection dose for M. natalensis. The animals died between 20 and 35 days after infection. After single infections with 50, 150, 400, or 800 eggs per animal the mortality of Mastomys challenged 36 or 52 days later was reduced to 0--30%. Using X-irradiated embryonated eggs for immunization only repeated administration led to protection in 70 to 80% of the animals. About 40% of the animals could be protected by the intraperitoneal injection of non-embryonated eggs. If death occurred it was delayed. The combination of X-irradiated stages and eggs did not enhance the protection.

Taenia crassiceps metacestode vesicular fluid antigens shared with the Taenia solium larval stage and reactive with serum antibodies from patients with neurocysticercosis.

After removal of host (mouse) serum proteins (albumin, transferrin, IgG and another five unidentified proteins) by immunoaffinity chromatography, the vesicular fluid of T. crassiceps metacestodes (TcVF) was immunoelectrophoretically examined for antigens recognized by rabbit antiserum to aqueous crude extract of T. solium cysticerci. Based on a precipitate pattern developed in electroimmunodiffusion, nine cross-reactive antigens could be demonstrated. In the ELISA, TcVF was shown to be a potent antigen for the demonstration of IgG antibodies in the sera of Mexican patients (n = 14) with confirmed neurocysticercosis (mean E490 values +/- SD: 0.39 +/- 0.38) although it was less sensitive when compared to T. solium VF (0.95 +/- 0.54). Sensitivity was much higher using cross-reactive TcVF antigens selected by immunoaffinity chromatography with rabbit IgG antibodies to larval T. solium crude extract (0.87 +/- 0.57). SDS-PAGE fluorograph of cross-reactive, radioiodinated TcVF protein antigens and selected by antibodies of individual neurocysticercosis sera (n = 13), exhibited six to nine bands depending on the serum tested. Altogether ten 125I-labelled proteins (Mr range from about 20,000 to 200,000) were recognized by neurocysticerosis antibodies. Four proteins (Mr about 22,000, 25,000, 32,000 and 45,000) were detected by all sera.