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.

Randomised clinical trial: the efficacy and safety of propionyl-L-carnitine therapy in patients with ulcerative colitis receiving stable oral treatment.

BACKGROUND: Ulcerative colitis (UC) is characterised by impaired fatty-acid oxidation; l-carnitine has a key role in fatty-acid metabolism and short-chain fatty acids such as butyrate and propionate are important energy source for intestinal epithelial cells. AIM: To evaluate efficacy and safety of colon-release propionyl-L-carnitine (PLC) in patients with mild-to-moderate UC receiving stable oral aminosalicylate or thiopurine therapy. METHODS: In a multicentre, phase II, double-blind, parallel-group trial, patients were randomised to receive PLC 1 g/day, PLC 2 g/day or placebo. Main inclusion criteria were as follows: age 18-75; disease activity index (DAI) score 3-10 inclusive, be under oral stable treatment with aminosalicylate or thiopurine. The primary endpoint was clinical/endoscopic response, defined as a decrease in DAI score ≥ 3 points or remission, defined as a DAI score ≤ 2 with no individual sub-score > 1. RESULTS: Of 121 patients who were randomised, 57 of 79 (72%) patients receiving PLC (combined 1 g and 2 g cohort) had a clinical/endoscopic response vs. 20 of 40 (50%) receiving placebo (P = 0.02). Specifically, in PLC 1 g/day group, 30 of 40 (75%) patients had clinical/endoscopic response (P = 0.02 vs. placebo) and 27 of 39 (69%) in the PLC 2 g/day group (P = 0.08 vs. placebo). Rates of remission were 22/40 (55%), 19/39 (49%), 14/40 (35%) in the PLC 1 g, PLC 2 g, and placebo groups, respectively. PLC had a similar safety profile to placebo; the most common adverse events were gastrointestinal. CONCLUSION: Propionyl-L-carnitine 1 g/day should be investigated further as a co-treatment for mild-to-moderate ulcerative colitis (NCT-01026857).

Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells.

Although new culture conditions enable homogeneous and long-term propagation of radial glia-like neural stem (NS) cells in monolayer and serum-free conditions, the efficiency of the conversion of NS cells into terminally differentiated, functionally mature neurons is relatively limited and poorly characterized. We demonstrate that NS cells derived from adult mouse subventricular zone robustly develop properties of mature neurons when exposed to an optimized neuronal differentiation protocol. A high degree of cell viability was preserved. At 22 days in vitro, most cells (65%) were microtubule-associated protein 2(+) and coexpressed gamma-aminobutyric acid (GABA), GAD67, calbindin and parvalbumin. Nearly all neurons exhibited sodium, potassium and calcium currents, and 70% of them fired action potentials. These neurons expressed functional GABA(A) receptors, whereas activable kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-D-aspartic acid receptors were present in approximately 80, 30 and 2% of cells, respectively. Antigenic and functional properties were efficiently and reliably reproduced across experiments and cell passages (up to 68). This is the first report showing a consistent and reproducible generation of large amounts of neurons from long-term passaged adult neural stem cells. Remarkably, the neuronal progeny carries a defined set of antigenic, biochemical and functional characteristics that make this system suitable for studies of NS cell biology as well as for genetic and chemical screenings.

[Vacuum-assisted breast biopsy for diagnosis of non-palpable lesions: experience with 226 cases]. / Biopsia mammaria "vacuum-assisted" per la diagnosi di lesioni non palpabili: descrizione dei primi 226 casi.

Vacuum-assisted breast biopsy (VABB) is now available for non-palpable lesions. The present study describes the results obtained from 226 consecutive VABBs performed at "L. Sacco" Hospital, Milan, from November 2005 to July 2007 (198 stereotactic and 28 ultrasonographic procedures). Adequate tissue samples for histopathological evaluation were obtained in 225 cases (99.6%). The diagnoses were as follows: 9 normal tissues (4%), 97 benign (43%), 25 "probably benign" (11%), 4 "suspicious for malignancy" (2%) and 90 malignant (40%, 53 in situ and 37 infiltrating carcinoma). Of the 90 malignant cases, 38 (42.2%) underwent subsequent surgical excision in our Unit; 84.2% (32/38) had concordant histopathological findings. In conclusion, VABB is an accurate and safe technique for diagnosis of non-palpable lesions, and in experienced hands avoids unnecessary surgical procedures.

Sonohysterographic endometrial sampling and hysteroscopic endometrial biopsy: a comparative study.

OBJECTIVES: To compare the quantity and quality of endometrial tissue sampled at saline contrast sonohysterography (SCSH) with that obtained by directed endometrial biopsy by operative hysteroscopy in patients with diffusely thickened and/or inhomogeneous endometrium at SCSH. A secondary aim was a comparison of the extent of procedure-related pain. METHODS: One hundred and twenty-eight patients with diffusely thickened (> 4 mm) and/or inhomogeneous endometrium at SCSH were prospectively recruited. Endometrial sampling was performed at the end of SCSH using the same 4.7-mm intrauterine catheter that had been used for saline instillation. These samples were compared to directed endometrial biopsies obtained with the guidance of an office 5-mm hysteroscope. After hysteroscopy, an extended guided curettage was performed under general anesthesia, providing specimens that were considered the gold standard for histological diagnosis. Endometrial specimen area (mm(2)), histologic concordance and procedure related pain (10-cm VAS) were compared for the two techniques. RESULTS: The median age of 88 pre- and of 40 post-menopausal patients was 41 (interquartile range, 34-48) years and 57 (interquartile range, 52-67) years, respectively. The median area of endometrial specimen obtained by SCSH was 25.1 (interquartile range, 12.4-52.3) mm(2) and was not significantly different from that obtained by hysteroscopy (16.9 (interquartile range, 10.0-52.7) mm(2)). The K values of the two different techniques for typical hyperplasia (n = 61) and for premalignant and malignant lesions (n = 26) were 0.91 and 0.94, respectively. Procedure-related pain was not significantly different between pre- and postmenopausal patients for both sampling techniques. CONCLUSIONS: SCSH with sampling proved to be as good as and as tolerable as hysteroscopic biopsy in cases with diffusely thickened and/or inhomogeneous endometrium. Both these imaging and biopsy techniques should be considered a reliable outpatient procedure in the management of patients with abnormal uterine bleeding. Published by John Wiley & Sons, Ltd.

Small bowel stenosis in Crohn's disease: clinical, biochemical and ultrasonographic evaluation of histological features.

AIM: To establish whether intestinal ultrasound, clinical or biochemical indices of activity can assess histological features of ileal stenosis in Crohn's disease. METHODS: In 43 patients undergoing surgery for a single ileal stenosis, clinical and biochemical parameters, as well as intestinal ultrasound, were assessed prior to surgery. The echo pattern of thickened bowel segments at the site of stenosis was classified as hypoechoic, stratified or mixed (segments with/without stratification). During surgery, stenoses were identified, resected and then histologically examined using standardized criteria. RESULTS: Clinical and biochemical indices of activity showed an overall weak positive correlation with histological inflammatory parameters and a negative correlation with fibrosis. The intestinal ultrasound echo pattern at the stenosis site was stratified in 25 patients, hypoechoic in 14 and mixed in four. Stenoses characterized by a stratified echo pattern showed a significantly higher degree of fibrosis, those characterized by hypoechoic echo pattern showed a higher degree of inflammation, while stenoses with a mixed echo pattern showed high degrees of both fibrosis and inflammation. CONCLUSION: Ultrasound and, to a lesser degree, clinical and laboratory indices discriminate between inflammatory and fibrotic ileal stenoses complicating Crohn's disease, thus allowing appropriate medical and/or surgical treatment to be defined.

Evolution of renal pathology in Fabry disease.

UNLABELLED: Fabry disease is a rare lysosomal storage disorder which results from deficient activity of the enzyme alpha-galactosidase A. The resultant deposition and progressive accumulation of glycosphingolipids in all types of body tissue leads to severe clinical manifestations involving the heart, CNS and kidney. Renal manifestations are observed relatively early in the course of the disease, and progression to end-stage renal failure is common in hemizygous males in the third to fifth decades of life. Renal biopsy specimens reveal evidence of diffuse intracytoplasmic glycosphingolipid accumulation, mainly affecting podocytes and epithelial cells of distal tubules, which are strikingly enlarged and vacuolated. On electron microscopy the deposits appear as typical osmiophilic inclusion bodies in the cytoplasm of all kinds of renal cells, and show a characteristic 'onion skin' or 'zebra' appearance. These pathological features are also evident in heterozygous females. Deposits occur before the development of renal impairment. As patients age, the disease progresses in cells throughout the kidney, and is associated with increasing glycosphingolipid accumulation. CONCLUSION: The age-related evolution of renal pathology in Fabry disease is closely correlated with progressive intracellular deposition of glycosphingolipid and ultimately leads to end-stage renal failure.

[Histopathological differential diagnosis in inflammatory bowel diseases]. / Diagnosi differenziali istopatologiche delle malattie infiammatorie intestinali.

In front of the suspicious diagnosis of an inflammatory bowel disease (IBD), the pathologist must have adequate and complete clinical, anamnestic, instrumental informations and, if possible, the previous histopathologic examinations. This is necessary because: the diagnosis of IBD is made with exclusion criteria, different pathologic entities may have similar macroscopic and microscopic findings and the characteristic lesions are often present in little number. The authors consider in this paper the problem of the differential diagnosis of IBD.

Six-drug sequential chemotherapy in non small cell lung cancer. A North Milan Group Study.

Forty-five patients with inoperable non small cell lung carcinoma were treated according to a sequential polychemotherapeutic regimen with cisplatin-vinblastine (A), cyclophosphamide-etoposide (B), and adriamycin-vincristine (C). Patients were evaluated every two cycles. Ten patients (22.2%) showed a partial response with a mean duration of 20 weeks, and mean survival of 50.8 weeks. It is remarkable that, among them, 6 patients (13.3%) lived over 12 months and three (6.6%) over 18 months. The mean survival for all patients was 35.7 weeks. Toxicity was acceptable and reversible.

Pharmacokinetic study of intravenous and oral idarubicin in cancer patients.

Idarubicin (4-demethoxydaunorubicin) is a new anthracycline analogue which lacks the methoxyl group at the C-4 position of the aglycone moiety. The present study was undertaken to investigate the pharmacokinetics and bioavailability of idarubicin in man. The drug was administered at 3-week intervals to six patients by both intravenous and oral routes. Doses used were 13-15 mg/m2 intravenous and 45 mg/m2 p.o. Plasma levels of unchanged idarubicin and of its metabolite idarubicinal were assayed by high performance liquid chromatography (HPLC). After intravenous administration the plasma levels of the unchanged drug declined very rapidly reaching the sensitivity limits of the analytical method (1-2 ng/ml) 24 h after dosing. Plasma levels of idarubicinal reached a peak of about 10 ng/ml within two hours then decreased very slowly with a plasma t1/2 of about 2.5 days. After the oral dose of 45 mg/m2, the plasma level patterns of both parent compound and the idarubicinal were roughly similar to those after 15 mg/m2 intravenous except for the obvious difference linked to the absorption of idarubicin. The absorption of oral idarubicin was rapid and, in terms of area under curve of the metabolite, the availability after oral administration can be estimated as about 30% of the dose. The urine findings reflected the plasma situation. The metabolite levels were much higher and longer lasting than those of the parent compound. Urinary recovery after intravenous (16% of the dose in four days) and oral administration (approximately 5% of the dose) confirmed the 30% absorption estimated on the basis of plasma levels.(ABSTRACT TRUNCATED AT 250 WORDS)

VP16-213 and cyclophosphamide in non oat cell bronchogenic carcinoma.

Thirty-two patients with by non oat cell bronchogenic carcinoma were admitted to a protocol including Cyclophosphamide (CTX) 1,000 mg/m2 i.v. day 1, VP16-213 200 mg/m2 p.o. day 1-3, every 3 weeks. Partial remissions were seen in 2 of 27 evaluable patients; 16 of 27 showed no change. Mean survival was 36.4 weeks, median survival was 38 weeks.

[Adrenal cortex extract in the supportive therapy of neoplasm patients]. / Un estratto di corteccia surrenale nella terapia di supporto dei pazienti neoplastici.

A high-title (2000 UCD) adrenal cortex extract was used in the support management of 30 neoplastic patients. Evaluation was based on performance status (PS) according to Karnofsky and symptom incidence before and after the treatment. PS improved in 30%. There was a marked reduction in asthenia and anorexia, with significant changes in headache, nausea, and vomiting. Blood pressure, heart rate, and the blood chemistry picture were not altered.

[Automatic method for determination of serum antistreptolysin]. / Metodo automatico di dosaggio delle antistreptolisine sieriche

The AA. describe a method of determination of serum antistreptolysin employing "Technicon" AutoAnalyzer, this method permitting the execution of 40 determinations in an hour, with a time of analysis of about 16 minutes. The procedure of analysis is very easy, with a very high selectivity of values; a variation coefficient of 5,3% and an accuracy index of 4,4% were found. Since the amount of serum employed is 50 mul this method appears to be particularly suitable in paediatry and in serological screening.