Improved therapeutic response by distinct timing of multiple heat treatments during interstitial radiation in the Dunning R3327 prostate tumor model

PURPOSE: As a continuation of a previous study showing the efficacy of a single local tumor heat treatment (LTH) in combination with interstitial radiation (IR) in the Dunning tumor system R3327 (subline AT1), we evaluated higher doses and/or lower dose rates with an extended time course of IR treatment, which allowed greater flexibility for LTH applications. METHODS: IR was carried out by the insertion of one removable 192Ir seed into the center of a R3327-AT1 tumor, transplanted s.c. into the distal thigh of Copenhagen rats. LTH (43.5 degrees C, for 35 min) varied from one treatment just before IR to multiple applications beginning at 0 h and repeated every 48 h or 72 h. RESULTS: The Dunning subline R3327-AT1 is a very thermoresistant tumor, which did not reveal any thermal response when heated up to 44.5 degrees C for 35 min. IR alone produced a delay in tumor growth, related to dose and dose rates of 18-53 cGy/h. During longer treatment times, a single LTH just before the IR was no more effective than IR alone. Thermoradiotherapy with multiple LTH treatments given at intervals of between 48 h and 72 h resulted in a clear increase in growth delay. Radiosensitization was highest in all dose-rate groups where LTH was applied every 72 h during a complete course of IR. CONCLUSIONS: These results demonstrate the importance of administering a sequence of multiple applications of LTH during continuous low-dose-rate irradiation and they substantiate our earlier findings, with shorter exposure times, where one LTH given every 72 h appeared to be most efficient in the combined treatment of the Dunning rat prostate tumor R3327-AT1.     

Analysis of the oxidative catabolism of retinoic acid in rat Dunning R3327G prostate tumors

Transforming growth factor-beta (TGF-beta) is a pleiotropic regulatory factor of tissue remodeling. Angiogenesis, a prerequisite of tissue repair and tissue expansion, is induced by TGF-beta in vivo, while proliferation and migration of cultured endothelial cells are inhibited by TGF-beta. Indirect mechanisms stimulating angiogenesis and modification of TGF-beta effects by cell-matrix interaction have been postulated to account for this paradigm. Because cellular behavior in tissue remodeling is decisively determined by cell-matrix interactions, which in turn is mediated via integrins, we investigated the effect of TGF-beta on matrix-dependent endothelial cell functions. Integrin expression of human dermal microvascular endothelial cells (HDMEC) was measured by Northern blot and fluorescence-activated cell sorter analysis after TGF-beta treatment and correlated to cell-matrix interactions, which were studied in a colorimetric cell attachment assay as well as the Boyden chamber chemotaxis assay. We found a cell-specific downregulation of integrin expression in HDMEC on the level of mRNA as well as on the cell surface. This effect correlated well with the reduction of integrin-dependent cell adhesion to several matrix proteins, in particular to fibronectin. Moreover, TGF-beta decreased fibronectin-induced chemotaxis of HDMEC. Thus, TGF-beta controls cell-matrix interaction of HDMEC by down-regulation of integrin expression. This effect of TGF-beta reflects direct and cell-specific control mechanisms on microvascular cells that may be critical for the coordinated process of angiogenesis requiring a balance of stimulatory and inhibitory factors.     

Anisotropy of radiance in tissue phantoms and Dunning R3327 rat tumors: radiance measurements with flat cleaved fiber probes

OBJECTIVE: To compare the prevalence of conduct problems (CP) according to level of urbanization and to determine which factors account for the potential difference in prevalence rates. METHOD: Study 1 used a questionnaire survey of a nationally representative sample of 10,462 Norwegian adolescents. Study 2 used a questionnaire survey of a representative sample of 1,346 adolescents living in Oslo. Self-reported CP included most DSM-III-R criteria for conduct disorder. RESULTS: CP rates were similar in all levels of urbanization, except for the only semimetropolitan city in the country, the capital Oslo, which had CP rates twice those of the rest of the country. This increase rate could not be explained by a series of commonly advocated explanations: family structure and parental practices, social network, socioeconomic status, integration in community activities, religious involvement, and race. However, involvement in "soft" drugs and associating with antisocial peers could explain the statistically differential rates. Furthermore, in the Oslo study, adolescents' CP did not vary according to density of population or region within the city. CONCLUSIONS: The results support previous studies showing increased rates of CP in urban areas. However, urbanization must pass a certain threshold before it has this effect. Moreover, the lack of support for commonly advocated explanations for the difference between urban and nonurban areas suggests that investigations specifically addressing potential explanations for this difference should be conducted. The results indicate that the increased rates of substance use in highly urbanized areas may account for the difference in CP rates by prolonging and aggravating CP.     

Cryosurgical ablation of the normal ventral prostate plus adjuvant does not protect Copenhagen rats from Dunning prostatic adenocarcinoma challenge

PURPOSE: We wished to determine if cryosurgical ablation of the normal ventral prostate of Copenhagen rats confers protective immunity against a subsequent challenge with Dunning R3327 MatLyLu prostatic adenocarcinoma. In human melanoma, tumor antigens have been characterized as normal cellular proteins. We reasoned that cryosurgical ablation of the normal prostate along with immunostimulatory adjuvants might release prostatic antigens to the immune system engendering an immune response and rendering rats immune to prostatic cancer cells. MATERIALS AND METHODS: On day 0, Copenhagen rats underwent cryosurgical ablation of the normal ventral prostate, cryosurgery and intraprostatic injection of Complete Freund's Adjuvant (CFA), CFA injection alone, or laparotomy alone. On day 21, animals received a subcutaneous challenge of MatLyLu tumor cells. Tumor dimensions were recorded at regular intervals by a single blinded investigator. RESULTS: Animals receiving cryosurgical ablation of the normal ventral prostate or intraprostatic CFA developed tumors more frequently than animals receiving laparotomy alone and the effect was statistically significant if animals received both cryosurgical ablation of the prostate and intraprostatic CFA (3 experiments, 1 x 10(4) MatLyLu cells), total number with tumors/total number challenged: laparotomy alone 3/17, cryosurgical ablation 7/17, cryosurgery plus CFA 10/16 (p = 0.013 versus laparotomy, Fisher's exact test), CFA alone 9/17. CONCLUSIONS: Cryosurgical ablation of the normal rat ventral prostate and intraprostatic CFA does not protect against and can enhance the tumorigenicity of MatLyLu prostatic cancer cells at distant sites. This could be occurring through specific immunologic effects or non-specific mechanisms induced by cryosurgery and CFA.     

Role of neutrophils and lymphocytes in inhibition of a mouse mammary adenocarcinoma engineered to release IL-2, IL-4, IL-7, IL-10, IFN-alpha, IFN-gamma, and TNF-alpha

Impressive inhibition of tumor growth has been observed after transduction of cytokine genes into tumor cells. Secreted cytokines do not affect the proliferation of a tumor directly but activate a host immune reaction strong enough to overcome its oncogenic capacity. However, the reaction mechanisms activated are difficult to interpret; because these mechanisms have been derived from experiments with different tumors, comparisons are hindered. To compare the reactive mechanisms induced by each cytokine, BALB/c mice were challenged with the parental cells of the syngeneic spontaneous mammary adenocarcinoma TSA, or with TSA cells engineered to release IL2, IL4, IL7, IL10, IFN alpha, IFN gamma, and TNF alpha, and the tumor growth area was studied histologically, ultrastructurally, and immunohistochemically. These observations were integrated with data on the growth and rejection patterns of TSA cells in mice depleted of natural killer (NK) cells, granulocytes, CD4+, or CD8+ lymphocytes. The rejection of TSA-IL2 and TSA-TNF alpha cells was associated with the massive presence of neutrophils, that of TSA-IL4 and TSA-IL7 cells with neutrophils and very small areas of colliquative necrosis, and that of TSA-IFN alpha and TSA-IL10 cells with extensive areas of ischemic-coagulative necrosis and some neutrophils. TSA-IFN gamma cells displayed a delay in growth, but were not rejected. Their growth areas comprised necrotic zones of ischemic necrosis devoid of neutrophils. The selective depletion experiments demonstrated that rejection of engineered TSA cells depends on several leukocyte populations. The weight of each population varied with the secreted cytokine, although neutrophils and CD8+ lymphocytes constantly played the major role. Employment of the same tumor line engineered with the genes of different cytokines showed that each cytokine evokes a distinct reaction and that tumor inhibition results from a complex mechanism in which neutrophils and CD8+ lymphocytes and ischemic necrosis are of primary importance.     

Cytogenetic and ploidy analysis of prostatic adenocarcinoma

The cytogenetic evaluation of prostatic adenocarcinoma has shown no consistent cytogenetic abnormalities. Despite manipulation of culture conditions, the majority of low-stage, untreated prostatic adenocarcinomas show a normal karyotype. We have performed cytogenetic analysis on eight primary prostate adenocarcinomas, using several control measures to increase the probability that any normal karyotype was derived from neoplastic cells rather than accompanying normal cells. Tumors were grown in media that encourages epithelial growth; DNA ploidy studies were performed before and after tissue culture; and immunohistochemical confirmation of the prostatic and epithelial nature of the cells was done following culture. Percentage of tumor on tissue sections adjacent to those submitted for culture was > 75% in all cases. Seven of eight cases were evaluable, and six cases showed no clonal abnormalities and were diploid. One tumor showed a population of tetraploid cells, without structural abnormalities. Three additional tumors showed evidence of tetraploidy by DNA analysis. One case showed nonclonal marker chromosomes and was aneuploid. This patient was pathologic Stage D. We conclude that the majority of prostatic adenocarcinomas at their inception may not show routinely detectable cytogenetic abnormalities. However, tetraploidy may play a role in the evolution of prostatic adenocarcinoma.     

The role of IL-4, IL-10, and TNF-alpha in the immune suppression induced by ultraviolet radiation

The two main catalytic residues Cys25 and His159 of the monomeric cysteine protease papain are located on different walls of a cleft formed by two domains. This topology suggests a possible relationship between relative domain organization and catalytic mechanism. The effect on enzymatic parameters of structural modifications at various locations of the two-domain interface of papain was examined by individual or double replacements by Ala of pairs of interacting residues. Most modifications had no effect on enzyme activity. However, the enzyme's substrate turnover (kcat) decreased following simultaneous alteration of the two most conserved residues, forming an apolar contact located 15 A away from the active site. The pH activity profile of the double mutant was unchanged, indicating a conserved ionization state of the active site thiolate-imidazolium ion pair. This state is strongly dependent on the distance separating the two residues, thus suggesting that the active site geometry has not been significantly altered. Efficient enzymatic activity in papain requires more than a correct active site geometry and is influenced by domain packing properties in a region remote from the active site.     

The contribution of individual variables to Hotelling's T2, Wilks' lambda, and R2

We examine the effect of each variable on the following statistics: the one-sample and two-sample Hotelling's T2, Wilks' lambda for multivariate analysis of variance, and R2 in multiple regression. For T2, the net effect of each variable is an increase in the multivariate statistic, and the particular factors determining the amount of increase are (i) the multiple correlation of the variable with all other variables, and (ii) how well the variable's contribution to falsifying the hypothesis can be linearly predicted from the other variables. The effect of each predictor variable on R2 is similar to the effect of each variable on T2. For Wilks' lambda, each variable induces a decrease, due to (i) the F for that variable alone, and (ii) the change in multiple correlation from within-sample to total-sample.     

Stimulation of IL-2 production and CD2R expression by splenopentin analogs

A multiplicity of peripheral nerve syndromes may develop in patients with diabetes mellitus, the commonest of which is a chronic symmetric sensory polyneuropathy, often associated with autonomic neuropathy. Once established, it is largely irreversible. Acute painful diabetic sensory neuropathy is a separate entity with a favorable prognosis. It now seems likely that chronic inflammatory demyelinating polyneuropathy occurs with greater frequency in diabetic subjects than in the general population and is one explanation for the occurrence of a predominantly motor polyneuropathy. Focal and multifocal peripheral nerve lesions are seen mainly in older diabetic patients and comprise cranial, thoracoabdominal and limb nerve lesions, the last including proximal lower limb diabetic motor neuropathy (diabetic amyotrophy). With this wide array of disorders and the frequency of diabetes, it is important to distinguish those that are directly or indirectly related to diabetes from those that have a coincidental relationship.     

Early IL-2/sIL-2R surge following surgery leads to temporary immune refractoriness

High serum level of immunoreactive but not biologically active IL-2 was detected 1 day after surgery in patients undergoing major operation (abdominal, open-heart), in proportion to the tissue injury caused by surgical trauma. IL-2 values were highest in those patients who underwent open-heart surgery and received blood transfusions. In all patients they declined in the third and fourth post-operative days. Elevated serum levels of soluble IL-2 receptors (sIL-2R) were already present 1 day after operation, and peaked in the third and fifth post-operative days after mitogen triggering. Blood lymphocytes derived from operated patients secreted reduced amounts of both IL-2 and sIL-2R compared with control lymphocytes. The extent and duration of this reduction were also proportional to the tissue trauma and were affected by blood transfusions. Based on these data we suggest that early post-operative systemic immunological activation (appearance of IL-2 in the serum) is followed by elevation of sIL-2R, which then interferes with IL-2-dependent immunity. Blood lymphocytes are probably not involved in the post-operative immunological activation. The trigger for and the site of IL-2/sIL-2R synthesis are not yet clear.     

Thioperamide, a histamine H3-receptor blocker, facilitates vasopressor response to footshocks

The predisposition of the newborns to contract infections diseases is dependent upon the limited efficiency of their immune mechanisms. Congenital infections amount to 5.7% in the research material, and the acquired infections 1.15%. The isolation of the microorganism is the basis for treating infections-the profiles of the pathogenic bacterial in flora were subjected to analysis. Im generalised infections Stafphylococcus epidermidis makes 56.6% and E. Coli accounts for 87.5 of the infections of the urinary system. In our research the late sepsis and pneumonia are more frequently the result of the hospital infection (14.2%) in the cases of congenital infections-pneumonia and the infection of the urinary system (72%). Hematologic indicators such as: leucopenia, thormbocytopenia, I/T are distinct infection markers (those were found in 31% of the cases). The CRP protein shows the lowest values in congenital infections, still monitoring its level is useful for assessing the effectiveness of the undertaken antybacterial treatment. The newborns of male sex (58%) more often prone to infection. Pneumonia is the manifestation pertaining to an organ in 70% of congenital infections, the infection of urinary system amounts to 17.1%.     

Telomeres, senescence and cellular radiation response

Telomeres shield the ends of chromosomes from degradation and end-to-end fusions. They shorten at each cell division and when they reach a critically short length, cells arrest in the G1 phase of the cell cycle and undergo senescence. This effectively limits the proliferative potential of cells. Senescence functions as a tumour suppressor mechanism and appears to contribute to the process of ageing. If senescence is circumvented by tumour viruses, proliferation is re-initiated until cells enter crisis. Activation of telomerase prevents telomere attrition and cells become immortal. Cellular response to ionizing radiation involves induction of cell cycle checkpoint arrests and programmed cell death. Because radiation produces double strand breaks in DNA, which cause telomere-less chromosome ends, radiation response appears to be the result of inappropriate induction of cellular senescence mechanisms.     

Bronchial reactivity to prostaglandins F2alpha, E2, and histamine in different types of asthma

Bronchial reactivity to prostaglandins F2alpha (PGF2alpha), E2 (PGE2) and histamine has been studied in 27 patients with aspirin-sensitive asthma and in 28 asthmatics without this sensitivity. Of the latter group, 13 patients had atopic, 9 infectious, and 6 mixed type of asthma. Atopic patients were characterized by vivid reactivity to low doses of both PGF2alpha and histamine. In patients with infectious asthma significantly higher doses of both PGF2alpha and histamine were necessary to induce bronchoconstriction as compared to atopics. Aspirin-sensitive patients responded quickly with bronchial spasm to similar doses of histamine as atopics, but tolerated significantly higher doses of PGF2alpha. There was no difference in reactivity to PGF2alpha between patients with aspirin sensitivity and those with infectious asthma. 5 and 10 min after administration of 60 microgram PGE2 significantly better improvement in ventilation occurred in aspirin-sensitive patients than in those of either atopic or infectious groups. The results obtained point to differences in bronchial reactivity to prostaglandins and histamine depending on type of asthma and severity of its symptoms.     

Seminal cytokine concentrations (IL-1beta, IL-2, IL-6, sR IL-2, sR IL-6), semen parameters and blood hormonal status in male infertility

Several lines of evidence indicate that cyokines are involved in male fertility. They are secreted by different parts of the male genital tract and may exert effects on steroidogenesis, spermatogenesis and sperm functions. We measured the concentrations of interleukins (IL-beta, IL-2, IL-6) and those of interleukin soluble receptors (sR IL-2, SR IL-6) in semen of fertile subjects (n = 21) and of patients with a range of andrological diseases (n = 119). The seminal concentrations of cytokines were analysed according to semen parameters as well as to the blood hormonal profiles of follicle stimulating hormone, luteinizing hormone and testosterone. An increase of IL-1beta was observed in the group of patients with infertility. No difference was found between the different subgroups defined on the basis of progressive motility, percentage of abnormal forms and diagnosis of infection. The seminal cytokine concentrations were independent of the blood hormonal status. Our data suggest that the determination of interleukins (-1beta, -2 and -6) or interleukin soluble receptors (sR IL-2, sR IL-6) in human spermatozoa does not provide convenient information in male routine infertility work-up.     

Rapid-fractionation preoperative chemoradiation, pancreaticoduodenectomy, and intraoperative radiation therapy for resectable pancreatic adenocarcinoma

PURPOSE: To evaluate the toxicities, radiographic and pathologic responses, and event-free outcomes with combined modality treatment that involves preoperative rapid-fractionation chemoradiation, pancreaticoduodenectomy, and electron-beam intraoperative radiation therapy (EB-IORT) for patients with resectable pancreatic adenocarcinoma. PATIENTS AND METHODS: Patients with radiographically resectable localized adenocarcinoma of the pancreatic head were entered onto a preoperative protocol that consisted of a 2-week course of fluorouracil (5-FU) 300 mg/m2 daily 5 days per week and concomitant rapid-fractionation radiation 30 Gy, 3 Gy daily 5 days per week. Radiographic restaging was performed 4 weeks after chemoradiation, and patients with localized disease underwent pancreaticoduodenectomy with EB-IORT 10 to 15 Gy. RESULTS: Thirty-five patients were entered onto the study and completed chemoradiation, 34 (97%) as outpatients. Three patients (9%) experienced grade 3 nausea and vomiting; no other grade 3 or 4 toxicities were observed. Of the 27 patients taken to surgery, 20 patients (74%) underwent pancreaticoduodenectomy with EB-IORT. All patients had a less than grade III pathologic response to preoperative chemoradiation. At a median follow-up of 37 months, the 3-year survival rate in patients who underwent combined modality therapy was 23%. CONCLUSION: Combined modality treatment with preoperative rapid-fractionation chemoradiation, pancreaticoduodenectomy, and EB-IORT is associated with minimal toxicity and excellent locoregional control. This represents one approach to maximize the proportion of patients who receive all components of combined modality therapy and avoids the toxicity of pancreaticoduodenectomy in patients found to have metastatic disease at the time of restaging.     

The response of lymph node metastases of testicular teratoma to radiation therapy

The age-dependent changes in reducible collagen cross-links, Schiff bases and their precursors were followed for human bone and articular cartilage throughout the whole life span from 3 to 89 years of age. With aging, the reduced Schiff bases dihydroxylysinonorleucine remarkable decreased, wheras hexitollysine peaks increased significantly in both collagens. Dihydroxynorleucine, the precursor of the cross-link, was found to increase slightly with age. But the changes in hydroxylysinonorleucine and lysinonorleucine were comparatively small. These changes were discussed in relation to the increased insolubility observed in both collagens at higher age.     

Increased response to antigen and histamine release in smaller sensitized canine bronchi

Presynaptic alpha 2-autoreceptors in mouse atria were characterized in terms of the alpha 2A, alpha 2B, alpha 2C and alpha 2D subtypes. Segments of the atria were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. The affinity of up to 16 antagonists for the autoreceptors was assessed as (1) pEC30% values. i.e. concentrations that increased previously autoinhibited release of 3H-noradrenaline (120 pulses, 3 Hz) by 30%, and (2) pKd values against the release-inhibiting effect of 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) under conditions of no or little autoinhibition (2 trains of 20 pulses, 50 Hz, train interval 120 s). The pKd values correlated well with the pEC30% values (r = 0.98; P < 0.001; slope of regression line 0.93), indicating that UK 14,304 and released noradrenaline modulated the release of noradrenaline through pharmacologically identical receptors. Comparison with antagonist affinities for (1) prototypic native alpha 2 radioligand binding sites, (2) radioligand binding sites in COS cells transfected with alpha 2 subtype genes, and (3) previously classified presynaptic alpha 2-autoreceptors-all taken from the literature-indicated that the mouse atrial autoreceptors corresponded to the alpha 2D subtype. For example, the pKd values at mouse atrial auto-receptors correlated closely with pKd values at native alpha 2D binding sites in the bovine pineal gland (r = 0.96; P < 0.001); with pKd values at alpha 2D binding sites in COS cells transfected with the rat alpha 2D gene (r = 0.85; P < 0.01); and with pKd values at guinea-pig cerebral and atrial and mouse cerebral alpha 2D-autoreceptors (r = 0.96-0.98; P < 0.001). The antagonist pKd values at mouse atrial autoreceptors correlated less with pKd values at alpha 2A, alpha 2B and alpha 2C sites. It is concluded that the presynaptic alpha 2-autoreceptors in mouse atria are alpha 2D. This identification supports the hypothesis that at least the majority of alpha 2-autoreceptors belong to the alpha 2A/D pair of orthologous alpha 2-adrenoceptors.