Comprehensive cancer predisposition testing within the prospective MASTER trial identifies hereditary cancer patients and supports treatment decisions for rare cancers.

BACKGROUND: Germline variant evaluation in precision oncology opens new paths toward the identification of patients with genetic tumor risk syndromes and the exploration of therapeutic relevance. Here, we present the results of germline variant analysis and their clinical implications in a precision oncology study for patients with predominantly rare cancers. PATIENTS AND METHODS: Matched tumor and control genome/exome and RNA sequencing was carried out for 1485 patients with rare cancers (79%) and/or young adults (77% younger than 51 years) in the National Center for Tumor Diseases/German Cancer Consortium (NCT/DKTK) Molecularly Aided Stratification for Tumor Eradication Research (MASTER) trial, a German multicenter, prospective, observational precision oncology study. Clinical and therapeutic relevance of prospective pathogenic germline variant (PGV) evaluation was analyzed and compared to other precision oncology studies. RESULTS: Ten percent of patients (n = 157) harbored PGVs in 35 genes associated with autosomal dominant cancer predisposition, whereof up to 75% were unknown before study participation. Another 5% of patients (n = 75) were heterozygous carriers for recessive genetic tumor risk syndromes. Particularly, high PGV yields were found in patients with gastrointestinal stromal tumors (GISTs) (28%, n = 11/40), and more specifically in wild-type GISTs (50%, n = 10/20), leiomyosarcomas (21%, n = 19/89), and hepatopancreaticobiliary cancers (16%, n = 16/97). Forty-five percent of PGVs (n = 100/221) supported treatment recommendations, and its implementation led to a clinical benefit in 40% of patients (n = 10/25). A comparison of different precision oncology studies revealed variable PGV yields and considerable differences in germline variant analysis workflows. We therefore propose a detailed workflow for germline variant evaluation. CONCLUSIONS: Genetic germline testing in patients with rare cancers can identify the very first patient in a hereditary cancer family and can lead to clinical benefit in a broad range of entities. Its routine implementation in precision oncology accompanied by the harmonization of germline variant evaluation workflows will increase clinical benefit and boost research.

Activation of murine macrophages and lymphocytes by Ureaplasma diversum.

Ureaplasma diversum is a pathogen in the bovine reproductive tract. The objective of the research was to study interactions with macrophages and lymphocytes which might elucidate aspects of pathogenetic mechanisms of this organism. We studied the activation of murine macrophages of C3H/HeN (LPS-responder) and C3H/HeJ (LPS-low-responder) genotype for TNF-alpha, IL-6, IL-1 and nitric oxide production and blastogenic response of C3H/HeJ splenocytes after Ureaplasma diversum stimulation. Live and heat-killed U. diversum induced TNF-alpha, IL-6 and IL-1 in peritoneal macrophage cultures of both C3H/HeN and C3H/HeJ mice in a dose dependent manner. Interferon-gamma modulated the cytokine production, by increasing the production of TNF-alpha, IL-6 and nitric oxide, but IL-1 secretion was only enhanced in C3H/HeJ macrophages stimulated by live ureaplasmas. Supernatant of U. diversum sonicate was mitogenic for murine spleen lymphocytes. The blastogenic response was dose dependent, and stimulation with both U. diversum and Concanavalin A seemed to have an additive effect. These results suggest that U. diversum, similar to other mycoplasmas, activates murine macrophages and lymphoid cells. The studies should be repeated with bovine cells in order to elucidate pathogenetic aspects of inflammation in cattle caused by U. diversum.

Functional interaction between local anaesthetics and calcium antagonists in guineapig myocardium: 1. Cardiodepressant effects in isolated organs.

Both local anaesthetics and calcium antagonists depress cardiac function. Therefore, we have studied the interaction of these compounds in the isolated myocardium of guineapigs. The negative inotropic effect of various local anaesthetics was investigated in left atria in the absence or presence of nitrendipine 10(-7) mol litre-1 (n = 7-8 in each group). In addition, the effect of bupivacaine was studied in the presence of several calcium antagonists. The factor by which the negative inotropic potency (EC50) of local anaesthetics was enhanced significantly in the presence of nitrendipine varied from mean 1.2 (SD 0.2) (benzocaine) to 3.0 (0.6) (bupivacaine). The EC50 of bupivacaine was lowered by all calcium antagonists. The potentiation factor varied from 1.4 (0.4) (verapamil) to 3.3 (0.6) (nifedipine). The effects of benzocaine (n = 12) and bupivacaine (n = 11) on the working heart were assessed either alone or after pretreatment with nifedipine 10(-8) mol litre-1. The effects of benzocaine on contractility remained unaltered in the presence of nifedipine, whereas the negative inotropic effect of bupivacaine increased significantly (for example, a 20% reduction in left ventricular maximum dP/dt occurred with bupivacaine 1.75 (0.16) x 10(-6) mol litre-1 alone compared with 1.1 (0.22) x 10(-6) mol litre-1 when combined with nifedipine). We conclude that the effects of some local anaesthetics, for example bupivacaine, on cardiac contractility are enhanced in the presence of calcium antagonists. The relevance of this interaction in patients remains to be determined.

The influence of casting methods on the surface microhardness and crystalline formation of aluminum-copper alloys.

The influence of air/gas, oxygen/gas, and electric casting methods on the surface microhardness and crystalline formation of three aluminum-copper alloys was verified. The inclusion, wax elimination, and casting of the pattern were accomplished by using conventional techniques for thermal expansion investment. After metalographic polishing, the surface hardness analysis was verified with a penetrator. The penetrations were transformed in Knoop hardness with the conversion table, and computed by variance analysis. The test sample surfaces were then etched with a reactive solution composed of equal parts of hydrogen peroxide and ammonium hydroxide. The crystalline grains precipitated by solution were photographed with Kodak film 32 ASA. The results demonstrated that the surface microhardness of the alloys was modified according to the heat sources used, and the crystalline grain disposition was also adversely influenced by the heat sources, with the exception of Goldent alloy.

An epizootic of Mycoplasma ovipneumoniae infection in captive Dall's sheep (Ovis dalli dalli).

In late spring of 1986, 10 of 23 Dall's sheep (Ovis dalli dalli) at the Metropolitan Toronto Zoo were moved to a new exhibit, where all developed severe respiratory signs refractory to anthelmintic and antibiotic therapy. In July, two animals died with chronic active bronch-pneumonia, and a third was euthanized because of pneumonia several months later. Bacteria were not isolated from the lungs of the first, steptococci and Pasteurella hemolytica were isolated from the other two, respectively; Mycoplasma ovipneumoniae was isolated from both. Pulmonary lesions in all three sheep were consistent with Mycoplasma sp. infection. Nasal swabs of the remaining animals yielded no consistent bacterial isolates; however, four of eight sheep were positive for M. ovipneumoniae. Viral cultures yielded an as yet unidentified herpesvirus. Sheep in the original and new herds had no serologic titers to parainfluenza-3, equine viral rhinopneumonitis, or infectious bovine rhinotracheitis, and had variable titers against bovine respiratory syncytial virus. No titers against M. ovipneumoniae were present in 13 sheep still in the original exhibit, but titers varied from 1:32 to 1:256 in eight pneumonic sheep. Sera taken from three sheep before or early in the outbreak were all negative for antibody to M. ovipneumoniae. Two of the affected Dall's sheep had been in contact with domestic sheep in the winter of 1985-1986, and M. ovipneumoniae was subsequently cultured from the domestic flock. Exposure to a new pathogen, and environmental and social stress in a new exhibit may have resulted in this severe disease in Dall's sheep.

Effect of sample freezing on the isolation of Mycoplasma spp. from the clitoral fossa of the mare.

The growth of Mycoplasma equigenitalium and Mycoplasma subdolum from specimens collected from the clitoral fossa of each of four Standardbred mares was not diminished by freezing of the specimens in liquid nitrogen (-196 degrees C) for up to 30 days when compared to samples cultured immediately.

Recovery of Mycoplasma spp. from the Reproductive Tract of the Mare during the Estrous Cycle.

The sites in the genital tract from which mycoplasmas could be recovered at various stages of the estrous cycle were studied in five Standardbred mares naturally infected with Mycoplasma. Mycoplasma equigenitalium and Mycoplasma subdolum were most frequently isolated from the clitoral fossa as compared to the vagina, cervix, and uterus. The lowest isolation prevalence was observed in the uterus. The recovery of Mycoplasma spp. from the clitoral fossa did not differ at any stage of the estrous cycle; however, recovery from the vagina, cervix, and uterus was variable during the cycle and more organisms were recovered on the day of ovulation than at any other time. From these results it was concluded that the clitoral fossa is the most likely "ecological niche" for Mycoplasma spp. in the mare. Ureaplasmas were not isolated.

Cellular inflammatory response in the lungs of calves exposed to bovine viral diarrhea virus, Mycoplasma bovis, and Pasteurella haemolytica.

Three, 5, or 7 days after inoculation with bovine viral diarrhea (BVD) virus (n = 12) or Mycoplasma bovis (n = 12), groups of calves were exposed to aerosols of Pasteurella haemolytica and were euthanatized 4 hours later. Histologic lesions in the lungs and the ratios of neutrophils to alveolar macrophages, collected by bronchoalveolar lavage, were compared with those of clinically healthy calves (n = 8) and calves inoculated with BVD virus only (n = 4), M bovis only (n = 4), or P haemolytica only (n = 2). Inoculation with BVD virus or M bovis did not have a significant (P greater than 0.05) effect on the neutrophil/macrophage ratio in the bronchoalveolar lavage. Aerosol exposure to P haemolytica induced a marked and significant (P less than 0.01) change in the neutrophil/macrophage ratio (from less than 1:9 to greater than 9:1). The reversed neutrophil/macrophage ratio in calves exposed to P haemolytica correlated well with the histologic changes in which small bronchi and bronchioles were plugged with purulent exudate. Inoculation with BVD virus did not induce gross or microscopic lesions in the lungs. Inoculation with M bovis resulted in a severe peribronchial lymphoid hyperplasia with mild exudation of neutrophils and macrophages into the cranioventral parts of the lungs.

Mycoplasma felis as a cause of pleuritis in horses.

Mycoplasma felis was the only organism recovered from the thoracic cavity of a horse with pleuritis. Large numbers of mildly degenerative neutrophils were in the pleural fluid. The horse developed a serologic response to M felis and recovered during hospitalization. Experimentally, a pony was inoculated in the thoracic cavity with a pure culture of the M felis isolate suspended in the pony's serum. A control pony was inoculated with serum only. Within 48 hours, the principal pony developed fever, increased respiratory rate, pleural effusion, and signs of pain. A highly cellular exudate with nondegenerative neutrophils and large numbers of M felis was recovered from the thoracic cavity. The control pony remained normal. The principal pony developed an antibody response to M felis. The control pony did not. Fourteen days after inoculation, both ponies were euthanatized. Necropsy revealed pleural inflammation in the principal pony. Pleural lesions were not found in the control pony.

An explanation for the water distribution in a hygroscopic expansion technique.

The water distribution in a controlled water addition technique was investigated using a Liquid Scintillation Counter. The mix water arrangement varied in different sections of the investment mass. The water distribution seemed to be the result of the influence of added water and a sprue base.

Phase identification of dental amalgam using transmission electron microscopy.

Dental amalgam specimens were polished and afterward etched by an acid iodine solution. The replicas were obtained by the vacuum coating technique. The electron micrographs indicated that it was possible to identify the silver-mercury (gamma1) and tin-mercury (gamma2) intermetallic compounds formed during amalgam setting.