Oncogenic CARMA1 couples NF-κB and ß-catenin signaling in diffuse large B-cell lymphomas.

Constitutive activation of the antiapoptotic nuclear factor-κB (NF-κB) signaling pathway is a hallmark of the activated B-cell-like (ABC) subtype of diffuse large B-cell lymphomas (DLBCL). Recurrent oncogenic mutations are found in the scaffold protein CARMA1 (CARD11) that connects B-cell receptor (BCR) signaling to the canonical NF-κB pathway. We asked how far additional downstream processes are activated and contribute to the oncogenic potential of DLBCL-derived CARMA1 mutants. To this end, we expressed oncogenic CARMA1 in the NF-κB negative DLBCL lymphoma cell line BJAB. By a proteomic approach we identified recruitment of ß-catenin and its destruction complex consisting of APC, AXIN1, CK1α and GSK3ß to oncogenic CARMA1. Recruitment of the ß-catenin destruction complex was independent of CARMA1-BCL10-MALT1 complex formation or constitutive NF-κB activation and promoted the stabilization of ß-catenin. The ß-catenin destruction complex was also recruited to CARMA1 in ABC DLBCL cell lines, which coincided with elevated ß-catenin expression. In line, ß-catenin was frequently detected in non-GCB DLBCL biopsies that rely on chronic BCR signaling. Increased ß-catenin amounts alone were not sufficient to induce classical WNT target gene signatures, but could augment TCF/LEF-dependent transcriptional activation in response to WNT signaling. In conjunction with NF-κB, ß-catenin enhanced expression of immunosuppressive interleukin-10 and suppressed antitumoral CCL3, indicating that ß-catenin can induce a favorable tumor microenvironment. Thus, parallel activation of NF-κB and ß-catenin signaling by gain-of-function mutations in CARMA1 augments WNT stimulation and is required for regulating the expression of distinct NF-κB target genes to trigger cell-intrinsic and extrinsic processes that promote DLBCL lymphomagenesis.

Anti-CD22 and anti-CD79B antibody drug conjugates are active in different molecular diffuse large B-cell lymphoma subtypes.

Antibody drug conjugates (ADCs), in which cytotoxic drugs are linked to antibodies targeting antigens on tumor cells, represent promising novel agents for the treatment of malignant lymphomas. Pinatuzumab vedotin is an anti-CD22 ADC and polatuzumab vedotin an anti-CD79B ADC that are both linked to the microtubule-disrupting agent monomethyl auristatin E (MMAE). In the present study, we analyzed the activity of these agents in different molecular subtypes of diffuse large B-cell lymphoma (DLBCL) both in vitro and in early clinical trials. Both anti-CD22-MMAE and anti-CD79B-MMAE were highly active and induced cell death in the vast majority of activated B-cell-like (ABC) and germinal center B-cell-like (GCB) DLBCL cell lines. Similarly, both agents induced cytotoxicity in models with and without mutations in the signaling molecule CD79B. In line with these observations, relapsed and refractory DLBCL patients of both subtypes responded to these agents. Importantly, a strong correlation between CD22 and CD79B expression in vitro and in vivo was not detectable, indicating that patients should not be excluded from anti-CD22-MMAE or anti-CD79B-MMAE treatment because of low target expression. In summary, these studies suggest that pinatuzumab vedotin and polatuzumab vedotin are active agents for the treatment of patients with different subtypes of DLBCL.

Some common errors of experimental design, interpretation and inference in agreement studies.

We signal and discuss common methodological errors in agreement studies and the use of kappa indices, as found in publications in the medical and behavioural sciences. Our analysis is based on a proposed statistical model that is in line with the typical models employed in metrology and measurement theory. A first cluster of errors is related to nonrandom sampling, which results in a potentially substantial bias in the estimated agreement. Second, when class prevalences are strongly nonuniform, the use of the kappa index becomes precarious, as its large partial derivatives result in typically large standard errors of the estimates. In addition, the index reflects rather one-sidedly in such cases the consistency of the most prevalent class, or the class prevalences themselves. A final cluster of errors concerns interpretation pitfalls, which may lead to incorrect conclusions based on agreement studies. These interpretation issues are clarified on the basis of the proposed statistical modelling. The signalled errors are illustrated from actual studies published in prestigious journals. The analysis results in a number of guidelines and recommendations for agreement studies, including the recommendation to use alternatives to the kappa index in certain situations.

Cell adhesion strength is controlled by intermolecular spacing of adhesion receptors.

Spatial patterning of biochemical cues on the micro- and nanometer scale controls numerous cellular processes such as spreading, adhesion, migration, and proliferation. Using force microscopy we show that the lateral spacing of individual integrin receptor-ligand bonds determines the strength of cell adhesion. For spacings > or = 90 nm, focal contact formation was inhibited and the detachment forces as well as the stiffness of the cell body were significantly decreased compared to spacings < or = 50 nm. Analyzing cell detachment at the subcellular level revealed that rupture forces of focal contacts increase with loading rate as predicted by a theoretical model for adhesion clusters. Furthermore, we show that the weak link between the intra- and extracellular space is at the intracellular side of a focal contact. Our results show that cells can amplify small differences in adhesive cues to large differences in cell adhesion strength.

Impact of receptor-ligand distance on adhesion cluster stability.

Cells in multicellular organisms adhere to the extracellular matrix through two-dimensional clusters spanning a size range from very few to thousands of adhesion bonds. For many common receptor-ligand systems, the ligands are tethered to a surface via polymeric spacers with finite binding range, thus adhesion cluster stability crucially depends on receptor-ligand distance. We introduce a one-step master equation which incorporates the effect of cooperative binding through a finite number of polymeric ligand tethers. We also derive Fokker-Planck and mean field equations as continuum limits of the master equation. Polymers are modeled either as harmonic springs or as worm-like chains. In both cases, we find bistability between bound and unbound states for intermediate values of receptor-ligand distance and calculate the corresponding switching times. For small cluster sizes, stochastic effects destabilize the clusters at large separation, as shown by a detailed analysis of the stochastic potential resulting from the Fokker-Planck equation.

Stability of adhesion clusters under constant force.

We solve the stochastic equations for a cluster of parallel bonds with shared constant loading, rebinding, and the completely dissociated state as an absorbing boundary. In the small force regime, cluster lifetime grows only logarithmically with bond number for weak rebinding, but exponentially for strong rebinding. Therefore rebinding is essential to ensure physiological lifetimes. The number of bonds decays exponentially with time for most cases, but in the intermediate force regime a small increase in loading can lead to much faster decay. This effect might be used by cell-matrix adhesions to induce signaling events through cytoskeletal loading.

Tap water burn prevention: the effect of legislation.

Five years after a 1983 Washington State law required new water heaters to be preset at 49 degrees C (120 degrees F), 77% of homes (84% of homes with postlaw and 70% of homes with prelaw water heaters) had tap water temperatures of less than 54 degrees C. In 1977, 80% of homes had tap water temperatures greater than 54 degrees C. Mean temperature in 1988 was 50 degrees C compared with 61 degrees C in 1977. Both changes were significant compared with 1977 temperatures. Few people increased their heater temperature after installation. Eighteen burn victims were identified from July 1979 through May 1988, for an average admission rate of 2.4 per year, compared with 5.5 per year in the 1970s. Compared with the 1970s, total body surface area burned, mortality, grafting, scarring, and length of hospital stay were all reduced, while the likelihood that scald burns were the result of child abuse increased to 50%. Both education campaigns and legislation have resulted in significantly safer water temperatures. Further, this change seems to have resulted in a reduction in frequency, morbidity, and mortality of tap water burn injuries in children. Lower water heater settings proved acceptable to the consumer.

[Factors influencing the result of kidney transplantation with special reference to the HLA system in conventional immunosuppression]. / Einflussfaktoren auf das Nierentransplantationsergebnis unter besonderer Berücksichtigung des HLA-Systems bei konventioneller Immunsuppression.

Among 331 recipients of cadaveric renal allografts, transplanted from 1976 to 1986 at the Rostock Transplant Center the graft survival rates have been analyzed. All patients have been treated by conventional immunosuppressive therapy using azathioprine and prednisolone. A relation between graft survival and immunological factors was found: the better the HLA match, the better the transplant results. The necessity of a restrictive transfusion regime was stressed. The higher the number of pretransplant transfusions, the higher is the panel reactivity with following lower graft survival. Cold ischemic time was without influence on graft survival. The introduction of cyclosporin may improve the graft survival rate as known of 10-20%.

[Retrograde ejaculation from the urologic viewpoint and the current status of therapeutic possibilities]. / Die retrograde Ejakulation aus urologischer Sicht und der gegenwärtige Stand der therapeutischen Möglichkeiten.

The successful treatment of ejaculatory disturbances is an unresolved problem even today, especially in the patient group of young men after retroperitoneal lymph node dissection because of testicular cancer. A promising therapeutic measure is the treatment with alpha-sympathicomimetics as Pryleugan and Gutron. Pryleugan is recommended as a first- line treatment, whereas Gutron is indicated after failure of this initial therapy. After unsuccessful medical treatment a procedure of sperm production, conservation and insemination is presented. A close cooperation between urologists and gynecologists is necessary for higher therapeutic success rates in ejaculatory disturbances.

Electron microscopic results on the ventral prostate of the rat after CdCl2 administration. A contribution towards etiology of the cancer of the prostate.

The prostate epithelium of rats which received repeated cadmium chloride (CdCl2) injections showed a gradual disruption of structural differentiation. Electron microscopy studies revealed severe changes in the ultrastructure affecting all epithelial cell organelles, particularly the nuclei, rough endoplasmatic reticulum, the golgi apparatus, and mitochondria. The cells infiltrating the stroma contained secretory vacuoles and their nuclear evaginations on the invasion front showed similarities to ultrastructural pathological changes in man. Examinations of the ventral prostate following oral CdCl2 administration revealed changes ranging in severity up to dysplasia, but there was no evidence of carcinoma.

[Relations between atypical hyperplasia, dysplasia and prostatic cancer and the significance of its appearance for the clinician]. / Beziehungen zwischen atypischer Hyperplasie, Dysplasie und Prostatakarzinom und die Bedeutung ihres Auftretens für den Kliniker.

In 89 preparations of patients with a carcinoma of the prostate and 100 preparations of patients with a hyperplasia of the prostate (adenoma of the neck of the bladder) the appearance of primarily atypical hyperplasias was investigated. In these cases in the carcinoma group in the uniform as well as in the pluriform carcinomas a coincidence of carcinoma and atypical hyperplasia of 55% was found. The coincidence is particularly high in adenocarcinomas and in carcinomas with dominating adenoid part, respectively, i. e. in more mature glandular structures. Among the various histological types of the atypical hyperplasia the microglandular/papillary clear-cellular form is dominating with 82%. In the group of the benign hyperplasia of the prostate gland a coincidence of the atypical hyperplasia of 33% was found. Though this rate, in comparison to other investigators, seems to be relatively high, it is still significantly lower than in the carcinoma. From this follows that the atypical hyperplasia, particularly its severest forms and dysplastic deformations from other hyperplasias (postatrophic hyperplasia, basal cell hyperplasia, atrophy and metaplasia of the prostate) now as ever are suspected that these cases in question are precancerous lesions. For the clinically working urologist the task is derived in such histopathological diagnoses by clinical and bioptical follow-up controls to find an already existing, but still unrecognized, or developing carcinoma of the prostate gland. In no case the diagnosis may cause surplus therapeutic consequences.

[Dielectric behavior of isolated dog kidneys following storage preservation and subsequent autotransplantation]. / Das dielektrische Verhalten isolierter Hundenieren nach Lagerungskonservierung und anschliessender Autotransplantation.

A measurement of the vitality of kidney transplants before the implantation is not yet possible, however, criteria for the proof of anoxic and hypothermic lesions on isolate kidneys are known. With the investigation means of the passive-electric behaviour over the determination of the DK-course and the dispersion of epsilon' an original evaluation of the vitality is possible. With the help of the measurement of epsilon' in frequencies between 0.5 MHz and 3 MHz we establish the proportion of intact cell membranes and mitochondria. The modul M3 explains the distribution of relaxing structures. Our hitherto performed experiments have shown that by means of the observation of the two sizes a judgment of the vitality seems to be possible. Examinations on bastard dogs of either sex were performed. For the supplementation of the dielectric measurements in certain time intervals tissue samples of the preserved kidneys were investigated electron-microscopically.

Carcinogenic effects of cadmium on the prostate of the rat.

The carcinogenic effects of cadmium were tested in an animal experiment to check the results of epidemiological studies showing that the incidence of prostatic carcinoma is above average in workers industrially exposed to high cadmium levels. For this purpose 1.9 mg CdCl2 (1.2 mg Cd/kg body weight) was injected into the right ventral prostatic lobe of 12-month-old male inbred rats of the Okamoto-toki strain. Simple hyperplasia was found in 38, atypical hyperplasia in 29, atypical hyperplasia with severe dysplasia in 11 and invasive prostatic carcinoma in 5 of 100 animals 270 days after the injection. Various mechanisms are discussed in connection with the carcinogenic action of cadmium.

[Radical prostatovesiculectomy in the treatment of prostate cancer. Indications, operative technic and personal results]. / Die radikale Prostatovesikulektomie in der Behandlung des Prostatakarzinoms. Indikation, Operationstechnik und eigene Ergebnisse.

Cancer of the prostate is the most common form of carcinoma in males of advanced age. If treated at an early stage, it can be healed by prostatovesiculectomy. Rectal prophylactic examination of all males aged 45 years or more should therefore be performed on a larger scale in order to detect these carcinomas as early as possible. Radical operation is indicated in males younger than 70 years old from stage T1 up to stages T3 and M0. Criteria vary in regard of the N-categories. The surgical technique practiced in the perineal and retropubic approach to prostatovesiculectomy is described, advantages and disadvantages are dealt with. No complete urinary incontinence could be observed in the 20 patients who underwent this operation at our hospital. Only 1 patient suffers from severe stress-induced incontinence, all others experiencing no difficulty in this respect. In 2 cases rectal lesions occurred during the operation. Postsurgical strictures occurring in the region of the urethra-bladder anastomosis in 2 other patients could be eliminated by internal urethrotomy. Impotence is the most common complication following this operation.

[Microsurgical blood vessel reconstruction in kidney transplantation]. / Mikrochirurgische Gefässrekonstruktion bei Nierentransplantationen).

Of 214 donor kidney transplants, 14.5% exhibited multiple arterial supply, 2.8%, injuries to vessels, and 1.8%, severance of the polar vessel. Microsurgical procedures were used on 14 of these kidneys; 11 functioned immediately, so that the patients could be discharged. Techniques of anastomosis using an operating microscope or magnifying spectacles are described.

[Therapeutic attitude of kidney transplantation patients--initial results of a psychological test study]. / Zur therapeutischen Haltung von Nierentransplantationspatienten--erste Ergebnisse einer testpsychologischen Studie.

After kidney transplantation the patient is subject to psychological stress due to problem situations. To date these problems have described only in very general clinical terms. Initial results of a psychological study on 26 kidney-transplant patients allow us to recognize factors which influence the patients' attitude to the disease and therapy. Such factors are: success of therapy, emotional tension, attitude towards the doctor and the self-confidence of the patient. The doctor-patient relation corresponds to the attitude towards therapy only in the immediate post-operative period and in patients with a negative attitude towards therapy. In the assessment of the doctor-patient relation it is only of medium importance whether the patient likes the doctor. The Study is being continued.