CDK12 promotes tumorigenesis but induces vulnerability to therapies inhibiting folate one-carbon metabolism in breast cancer.

Cyclin-dependent kinase 12 (CDK12) overexpression is implicated in breast cancer, but whether it has a primary or only a cooperative tumorigenic role is unclear. Here, we show that transgenic CDK12 overexpression in the mouse mammary gland per se is sufficient to drive the emergence of multiple and multifocal tumors, while, in cooperation with known oncogenes, it promotes earlier tumor onset and metastasis. Integrative transcriptomic, metabolomic and functional data reveal that hyperactivation of the serine-glycine-one-carbon network is a metabolic hallmark inherent to CDK12-induced tumorigenesis. Consistently, in retrospective patient cohort studies and in patient-derived xenografts, CDK12-overexpressing breast tumors show positive response to methotrexate-based chemotherapy targeting CDK12-induced metabolic alterations, while being intrinsically refractory to other types of chemotherapy. In a retrospective analysis of hormone receptor-negative and lymph node-positive breast cancer patients randomized in an adjuvant phase III trial to 1-year low-dose metronomic methotrexate-based chemotherapy or no maintenance chemotherapy, a high CDK12 status predicts a dramatic reduction in distant metastasis rate in the chemotherapy-treated vs. not-treated arm. Thus, by coupling tumor progression with metabolic reprogramming, CDK12 creates an actionable vulnerability for breast cancer therapy and might represent a suitable companion biomarker for targeted antimetabolite therapies in human breast cancers.

Apheresis treatment of cryoglobulinemic vasculitis: A multicentre cohort study of 159 patients.

OBJECTIVE: To assess the effectiveness of apheresis therapy (AT) in treating the clinical manifestations of patients with complicated cryoglobulinemic vasculitis (CV). METHODS: A retrospective cohort study of 159 CV patients attending 22 Italian Centers who underwent at least one AT session between 2005 and 2015. The response to AT was evaluated on the basis of a defined grading system. RESULTS: Peripheral neuropathy was the most frequent clinical condition leading to AT. Therapeutic plasma exchange was used in 70.4% of cases. The outcome of AT was rated very good in 19 cases, good in 64, partial/transient in 40, and absent/not assessable in 36. Life-threatening CV-related emergencies and renal impairment independently correlated with failure to respond to AT. The independent variables associated with an increased risk of death were age at the time of the first AT session, multi-organ life-threatening CV, the presence of renal impairment and failure to respond to AT. The time-dependent probability of surviving until CV-related death in the second year was 84%, with an AHR in patients with absent/not assessable response to AT of 11.25. CONCLUSION: In this study AT is confirmed to be a safe procedure in patients with CV. Early AT should be considered in patients with severe CV, especially in cases with impending renal involvement, in order to prevent irreversible kidney damage. Although its efficacy in patients with multi-organ failure is limited, AT is the only treatment that can rapidly remove circulating cryoglobulins, and should be considered an emergency treatment.

Distress and job satisfaction after robbery assaults: a longitudinal study.

BACKGROUND: External/intrusive violence at work can result in psychological distress and can be an important risk to employee health and safety. However, the vast majority of workplace violence studies have employed cross-sectional and correlational research, designed to examine immediate reactions after being assaulted at work. AIMS: To explore whether exposure to robbery as a traumatic event may contribute to the onset of typical symptoms of psychological distress (anxiety depression, dysphoria and loss of confidence) and job dissatisfaction over time. METHODS: We collected data by using a two-wave panel design, in which employees working the days of bank robberies, in an Italian bank, filled in a questionnaire between 48h and 1 week after the robbery (T1) and 2 months after the robbery (T2). We performed structural equation models to evaluate the fit of different models to our data. RESULTS: There were 513 participants at T1 (58% women) and 175 (34%) participants at T2 (62% women). There was a simultaneous association in which psychological distress leads to job dissatisfaction both following robbery and 2 months later. CONCLUSIONS: Our findings support a synchronous effects model and suggest that interventions after suffering physical assaults, apart from helping employees to recover their health, should consider restoring their trust and confidence in the organization. This study contributes to understanding the dynamic relationships between a robbery at work and its outcomes over time, by addressing several methodological deficiencies in previous longitudinal studies.

Fibronectin and cryoprecipitation.

Fourteen patients with essential mixed cryoglobulinemia were studied. The relevance of fibronectin (FN) in cryoprecipitation seems to be of relative importance since only two thirds of the cryoprecipitates have a higher cryocrit level when FN is present. On the other hand, the other samples show a lower precipitating activity after plasma heating at 60 degrees C. This reduction may reflect either a structural FN modification by heating or the denaturation of other cryoprecipitating molecules. It is possible, even if not necessary, that at least in vitro FN plays a role in the cryoprecipitate formation.

Efficacy of discontinuous flow centrifugation compared with cascade filtration in Waldenstrom's macroglobulinemia: a pilot study.

In the treatment of Waldenstrom's macroglobulinemia (WM) and lipoprotein or immune complex mediated diseases, centrifugal and membrane plasma separation systems have been successfully employed over the past years. More recently, semiselective double filtration systems have been used in isolated cases. While it is hoped that cascade filtration (CF) will soon become a routine technique, a note of caution came from some investigators because of the lack of any evidence that CF does really deplete patients' plasma of known pathogenic macromolecules. The aim of this study was to test the efficacy of CF in the removal of IgM paraproteins in comparison with discontinuous flow centrifugation (DFC). Eight patients were treated by DFC and seven by CF; there were no significant differences between the two groups in terms of sex, age, clinical severity and IgM plasma concentration. After DFC or CF sessions, IgM concentration diminished by 50% and plasma viscosity by 60%, producing a comparable mean reduction in patients' clinical severity. Our in vivo results thus preliminarily answer the question whether CF is capable of removing known pathogenic macromolecules from patients' plasma, and the technique appears to warrant further investigations.

Biochemical and clinical evaluation of a new cellulose diacetate secondary filter for cascade filtration.

We have performed 24 cascade filtration treatments in 8 patients with hyperviscosity syndrome (2 cases), essential mixed cryoglobulinemia, post-hepatitic cryoglobulinemia, Sjogrens disease, rheumatoid vasculitis, Miller-Fisher syndrome and chronic dysimmune polyneuropathy. New cellulose diacetate filters were employed, giving a satisfactory performance. At 1.5 L plasma treatment, the rejection rate for macromolecular plasma components was close to 90%, whereas albumin recovery was close to 70%. Treatments were clinically effective, confirming that cascade filtration is an alternative to conventional plasma exchange in patients with IgM or immune complex mediated diseases.

A critical approach to therapeutic apheresis in the management of inflammatory dysimmune polyneuropathies.

Data concerning 37 patients with inflammatory dysimmune polyneuropathy treated by discontinuous flow centrifugation, membrane plasma separation and cascade filtration are presented. Plasmapheresis was combined with immunosuppressants in patients with chronic or relapsing neuropathy (8 patients), cryoglobulinemic (6 patients) and paraneoplastic disease (2 patients), whereas 21 patients with acute Guillain-Barré syndrome (GBS) underwent exclusively plasmapheresis. Most patients were treated during the onset or progression of their disease and excellent or satisfactory clinical results were obtained in 18 patients with GBS, 6 with cryoglobulinemia, 2 with paraneoplastic disease and 4 with chronic relapsing polyneuropathy. Prior to therapy, 34 patients had high levels of immune complexes (CIC); this level was clearly reduced by plasmapheresis and clinical results correlated well with this removal. 3 patients with chronic dysimmune polyneuropathy, without any evidence of CIC, were completely unaffected by treatment. The possible role of CIC in demyelinating polyneuropathies is discussed on the basis of information given by cascade filtration treatment of 7 patients.

Plasmapheresis combined with lymphocytapheresis and cytotoxic drugs as a therapeutic modality in neuroimmunological diseases.

Rebound after plasmapheresis is thought to be due to enhanced antibody and/or immune complex production. To prevent rebound a combination of steroids and immunosuppressive drugs has been used: lymphocytapheresis was employed in most patients, for not more than 10 sessions. 50 patients with myasthenia gravis, inflammatory myopathy, chronic dysimmune polyneuropathy and immune complex polyneuropathy have been treated and long-lasting benefits obtained in 35 patients. Rebound effects were observed only when cytotoxic drugs were not given or discontinued too soon. 8 patients who had been treated by plasmapheresis combined with steroids alone, after some recurrences of their disease, were switched to cytotoxic drugs, steroids and lymphocytapheresis combined with plasma exchange: in this group 7 patients eventually gained long lasting remissions. Our clinical experience strongly supports the hypothesis of a synergy between plasmapheresis and immuno-suppressive measures.

Plasmapheresis and cytotoxic drugs for mixed cryoglobulinemia.

Fifteen patients with symptomatic cryoglobulinaemia were subjected to apheretic treatment when acute renal insufficiency, glomerulonephritis, severe generalized vasculitis and polyneuropathy unresponsive to conventional therapy or complications due to steroids, such as vertebral collapse, peptic ulcer and steroid diabetes, had appeared. Treatment was performed by discontinuous flow centrifugation or cascade filtration: when discontinuous flow centrifugation was employed, a mixture of saline, gelatin and fresh frozen plasma was used for replacement. Cytotoxic drugs were administered to patients with lymphoma (4 patients) or chronic active hepatitis (5 patients) and also to patients suffering from essential mixed cryoglobulinaemia. Exchanges were organized into courses of 3 to 5 sessions over 5 to 10 days and employed as a supportive measure. No patient underwent long-term treatment. A complete resolution of kidney damage, skin involvement and neurologic signs was observed when treatment was started early in the course of the disease, whereas unequivocal but moderate improvement was obtained in the case of long-lasting symptoms such as polyneuropathy. Relapses were seen in most patients when cytotoxic drugs had been discontinued abruptly. In 8 patients the solubility of cryoglobulins was studied by a recently developed turbidimetric assay. Following treatment the solubility increased; when solubility decreased, 2 patients of this group had a relapse. On the basis of these preliminary observations it appears that the possibility of predicting relapsing disease or the need of continuing therapy can eventually be achieved.

Cascade filtration: clinical application in 26 patients with immune complex and IgM mediated diseases.

A new technique which allows lymphocytapheresis to be combined with cascade filtration (CF) is described in this paper. This therapeutical approach was applied for the treatment of patients affected by necrotizing vasculitis (1), inflammatory myopathies (5), Cryoglobulinemia (5), immune complex polyneuropathies (7), rheumatoid arthritis (3) and psoriasis (3 patients). Two cases of Waldenstrom's macroglobulinemia were also treated after the onset of the hyperviscosity syndrome. 78 procedures have been performed without any untoward effect. From a clinical point of view all patients had some improvement following treatment, thereby confirming not only the clinical safety of this therapeutical approach but also its effectiveness at least in the management of diseases which usually respond to plasma exchange treatment. Laboratory investigations showed that with CF it is possible to selectively remove IgM, immune complexes, fibrinogen, lipoproteins and high molecular weight plasma components, sparing most albumin and IgG globulins (85 and 71%, respectively).

Evaluation of a new filter for membrane plasma separation (Preliminary observations).

A new plasma filter for membrane plasma separation was used in 10 patients with the following diagnoses: relapsing Guillain-Barré syndrome (one patient), multiple sclerosis (one patient), myasthenia gravis (two patients), cryoglobulinemia (two patients), rheumatoid arthritis (one patient) and psoriasis (three patients). In all patients but one the plasma flow was from 975 to 1,370 ml/h; extremely high levels of cryoglobulins were the cause of filter plugging during a single procedure. The composition of the effluent plasma was not significantly different from the patients' plasma as regards protein electrophoresis and the levels of immune complexes, immunoglobulins and complement. No platelet activation or blood clotting within the processed blood was observed during 20 procedures. No elevation of lysozyme levels or changes in the osmotic fragility of the red blood cells were found, confirming the excellent biocompatibility of these new filters. The clinical results were consistent with the volumes of plasma exchanged and the clinical diagnoses.

A method for the study of cryoglobulin solubilization curves at 37 degrees C. Preliminary studies and application to plasma exchange in cryoglobulinemic syndromes.

A simple method for the study of kinetic solubility curves of cryoglobulins is presented. In its first application to the study of 21 patients with cryoglobulinemia, it was possible to ascertain that clinical condition roughly correlates with decreased solubility whereas no correlation is found with per cent cryocrit. In the group of patients we studied, 6 underwent plasma exchange treatment when glomerulonephritis, acute renal insufficiency, cerebritis and polyneuropathy appeared: in these patients, following 2 to 5 apheretic sessions, solubility increased showing a sort of correlation with clinical benefits determined by treatment. The preliminarity of this study is underlined.

A simple and rapid test for the quantification of osmotic fragility of red blood cells.

We have developed a simple and rapid nephelometric test for the quantification of the osmotic fragility of red blood cells. 10 microliters of whole blood are mixed with hypotonic NaCl solutions under mechanical stirring while the hemolysis curve is continuously recorded for 1 min. In our preliminary studies, this test was reproducible, time-saving, easy to set up and to standardize. No false-positive or false-negative results have been recorded so far. Moreover, it seems that this test can differentiate heterozygous beta-thalassemic patients from patients with sideropenic anemia, which was hardly possible with previously proposed assays.

Plasma-exchange in neurological diseases.

This paper summarizes experience with plasma-exchange therapy for neurological diseases at the Saronno hospital. Most treatments were performed by discontinuous flow centrifugation, but membrane plasma separation and cascade filtration were also employed. Eighty-five patients with demyelinating diseases of the peripheral nervous system (Guillain-Barré syndrome, immune complex polyneuropathies, paraneoplastic polyneuropathies), demyelinating diseases of the central nervous system (multiple sclerosis, subacute sclerosing panencephalitis), dermatopolymyositis and myasthenia gravis have been treated so far. Particular attention is paid to the combination of plasmapheresis with lymphocytapheresis and immunosuppressive drugs. This therapeutic approach appears to bring about dramatic and sustained improvement in most patients with neurological diseases, thus altering their natural course.

Management of immune-mediated and paraproteinemic diseases by membrane plasma separation and cascade filtration.

Membrane plasma separation was utilized in the management of 19 patients with autoimmune, immune complex or paraproteinemic diseases. In these patients 34 procedures were carried out employing a new filter composed of polyvinyl alcohol hollow fibers with a nominal pore size of 0.4 micrometer. Cascade filtration was performed in 23 procedures using a second filter built up with ethylenevinylalcohol copolymer hollow fibers with a nominal pore size of 0.1-0.2 micrometer. Both filters employed showed a very good biocompatibility, and no untoward effect was observed. Using the differential sieving effect of cascade filtration it was possible to eliminate circulating immune complexes and immunoglobulin M without the loss of necessary plasma components.