Identification of OAT1/OAT3 as Contributors to Cisplatin Toxicity.

Cisplatin is among the most widely used anticancer drugs and known to cause a dose-limiting nephrotoxicity, which is partially dependent on the renal uptake carrier OCT2. We here report a previously unrecognized, OCT2-independent pathway of cisplatin-induced renal injury that is mediated by the organic anion transporters OAT1 and OAT3. Using transporter-deficient mouse models, we found that this mechanism regulates renal uptake of a mercapturic acid metabolite of cisplatin that acts as a precursor of a potent nephrotoxin. The function of these two transport systems can be simultaneously inhibited by the tyrosine kinase inhibitor nilotinib through noncompetitive mechanisms, without compromising the anticancer properties of cisplatin. Collectively, our findings reveal a novel pathway that explains the fundamental basis of cisplatin-induced nephrotoxicity, with potential implications for its therapeutic management.

Genome-Wide Study Links PNPLA3 Variant With Elevated Hepatic Transaminase After Acute Lymphoblastic Leukemia Therapy.

Remission induction therapy for acute lymphoblastic leukemia (ALL) includes medications that may cause hepatotoxicity, including asparaginase. We used a genome-wide association study to identify loci associated with elevated alanine transaminase (ALT) levels after induction therapy in children with ALL enrolled on St. Jude Children's Research Hospital (SJCRH) protocols. Germline DNA was genotyped using arrays and exome sequencing. Adjusting for age, body mass index, ancestry, asparaginase preparation, and dosage, the PNPLA3 rs738409 (C>G) I148M variant, previously associated with fatty liver disease risk, had the strongest genetic association with ALT (P = 2.5 × 10-8 ). The PNPLA3 rs738409 variant explained 3.8% of the variability in ALT, and partly explained race-related differences in ALT. The PNPLA3 rs738409 association was replicated in an independent cohort of 2,285 patients treated on Children's Oncology Group protocol AALL0232 (P = 0.024). This is an example of a pharmacogenetic variant overlapping with a disease risk variant.

Tumor-derived inducible heat-shock protein 70 (HSP70) is an essential component of anti-tumor immunity.

The anti-apoptotic function and tumor-associated expression of heat-shock protein 70 (HSP70) is consistent with HSP70 functioning as a survival factor to promote tumorigenesis. However, its immunomodulatory activities to induce anti-tumor immunity predict the suppression of tumor growth. Using the Hsp70.1/3(-/-)(Hsp70(-/-)) mouse model, we observed that tumor-derived HSP70 was neither required for cellular transformation nor for in vivo tumor growth. Hsp70(-/-) murine embryonic fibroblasts (MEFs) were transformed by E1A/Ras and generated tumors in immunodeficient hosts as efficiently as wild-type (WT) transformants. Comparison of Bcr-Abl-mediated transformation of WT and Hsp70(-/-) bone marrow and progression of B-cell leukemogenesis in vivo revealed no differences in disease onset or survival rates, and Eµ-Myc-driven lymphoma in Hsp70(-/-) mice was phenotypically indistinguishable from that in WT Eµ-Myc mice. However, Hsp70(-/-) E1A/Ras MEFs generated significantly larger tumors than their WT counterparts in C57BL/6 J immune-competent hosts. Concurrent with this was a reduction in intra-tumoral infiltration of innate and adaptive immune cells, including macrophages and CD8(+) T cells. Evaluation of several potential mechanisms revealed an HSP70-chemokine-like activity to promote cellular migration. These observations support a role for tumor-derived HSP70 in facilitating anti-tumor immunity to limit tumor growth and highlight the potential consequences of anti-HSP70 therapy as an efficacious anti-cancer strategy.

Is gait variability reliable? An assessment of spatio-temporal parameters of gait variability during continuous overground walking.

The assessment of gait variability has become an important indicator for quantifying motor performance. However, the use of treadmills is known to influence the temporal rhythm of gait, while non-continuous (i.e. stop-start) overground walking alters gait variability, leading to erroneous results. Through establishing the "8-walk", an overground walking protocol that allows the collection of a high number of consecutive gait cycles, the aim of this study was to determine the conditions under which gait variability can be assessed reliably. Twelve healthy subjects performed continuous barefoot walking at their preferred speed in a path shaped as an "8". Kinematic data of the dominant foot was collected while subjects walked along the straight 10 m sections of the 8-walk during sessions on two different days. Mean spatio-temporal parameters of gait and gait variability were computed for 10, 20, 30, 40, 50 and 60 consecutive cycles. All mean parameters of gait showed excellent reliability (ICC: 0.88-0.98) with only 10 cycles included in the analysis. However, the reliability of spatial and temporal parameters of gait variability improved with increasing number of cycles (ICC: 0.60-0.90) but levelled-off after 50 consecutive cycles, revealing an inter-day test-retest variability of ≈ 13%. To reliably assess gait variability and evaluate human motor performance, we propose the collection of at least 50 cycles and the use of an 8-walk protocol, which avoids the limitations of treadmill and non-consecutive walking protocols.

Microsporogenesis in Paspalum conspersum Schrad. (Virgata group) with different ploidy levels.

Knowledge about the cytology and reproductive behavior of a species is indispensable for hybridization programs. This is especially true for species belonging to the genus Paspalum, among which apomixis and a wide range of ploidy levels are frequently found. Paspalum conspersum Schrad. is a robust and warm-season perennial bunchgrass native to South America. Previous studies have indicated that both tetraploid and hexaploid races exist in this species; however, only information related to tetraploids has been applied to another taxon. In this study, a cytological investigation in two Brazilian accessions collected in different regions revealed tetraploidy in the accession BRA-012823 (2n = 4x = 40), with chromosome pairing in bivalents and normal meiosis and tetrad formation, and pentaploidy (2n = 5x = 50) in the accession BRA-022748, which presented total asynapsis. In this latter accession, 50 univalents could be scored at diakinesis. After alignment at the metaphase plate, sister chromatids segregated to the poles. Only one meiotic division (equational) occurred, and after cytokinesis, 100% of the dyads that formed had 2n microspores. The meiotic behavior during microsporogenesis, which showed 10 delayed univalents to reach the metaphase plate, suggests that this accession is a recent natural hybrid constituted by a parental genome with 40 chromosomes and another with 10 chromosomes. The potential usage of these accessions in Paspalum breeding has been discussed.

Genetic deletion of caspase-2 accelerates MMTV/c-neu-driven mammary carcinogenesis in mice.

Despite being the most evolutionarily conserved of the mammalian caspases, little is understood about the cellular function of caspase-2 in normal tissues or what role caspase-2 may have in the progression of human disease. It has been reported that deletion of the caspase-2 gene (Casp2), accelerates Eµ-myc lymphomagenesis in mice, and thus caspase-2 may act as a tumor suppressor in hematological malignancies. Here, we sought to extend these findings to epithelial cancers by examining the potential role of caspase-2 as a tumor suppressor in the mouse mammary carcinogenesis model; MMTV/c-neu. The rate of tumor acquisition was significantly higher in multiparous Casp2(-/-)/MMTV mice compared with Casp2(+/+)/MMTV and Casp2(+/-)/MMTV mice. Cells from Casp2(-/-)/MMTV tumors were often multinucleated and displayed bizarre mitoses and karyomegaly, while cells from Casp2(+/+)/MMTV and Casp2(+/-)/MMTV tumors never displayed this phenotype. Tumors from Casp2(-/-)/MMTV animals had a significantly higher mitotic index than tumors from Casp2(+/+)/MMTV and Casp2(+/-)/MMTV animals. Cell cycle analysis of Casp2(-/-) E1A/Ras-transformed mouse embryonic fibroblasts (MEF) also indicated a higher proliferative rate in the absence of caspase-2. In vitro assays further illustrated that MEF had increased genomic instability in the absence of caspase-2. This appears to be due to disruption of the p53 pathway because we observed a concomitant decrease in the induction of the p53 target genes, Pidd, p21 and Mdm2. Thus caspase-2 may function as a tumor suppressor, in part, through regulation of cell division and genomic stability.

The novel carbohydrate tumor antigen C2-O-sLe x is upregulated in canine gastric carcinomas.

Sialyl Lewis x-modified core 2 branched O-glycans (C2-O-sLe(x)) on human leukocytes mediate much higher-affinity adhesion to selectins on activated vascular endothelium than does sialyl Lewis x on other structures. In some canine and human carcinomas, high expression of sLe(x)-decorated carbohydrates has been associated with metastasis and, in humans, a poor prognosis, but detection in canine gastric carcinomas is unreported. The authors hypothesized that these carbohydrates are highly expressed in more malignant types of canine gastric carcinomas, they promote metastasis, and they are associated with a poorer prognosis for dogs. The objectives were to determine the presence and importance of C2-O-sLe(x) expression in canine gastric carcinomas. Routine histological sections of 16 canine gastric carcinomas were categorized on the basis of 3 classification schemes: World Health Organization, Lauren, and Goseki. Serial sections were stained with antibodies directed against C2-O-sLe(x) (CHO-131 monoclonal antibody), cytokeratin (Lu-5 monoclonal antibody), and stains to detect neutral and acid mucins (periodic acid-Schiff and alcian blue). Whereas normal gastric mucosal epithelial cells were negative for C2-O-sLe(x), 56% of the tumors examined were positive for C2-O-sLe(x). Importantly, the majority of more poorly differentiated tumor types had more numerous and larger intensely stained areas of C2-O-sLe(x) expression compared with moderate to well-differentiated tumor types. Signet ring-type carcinomas had markedly higher distribution and intensity of periodic acid-Schiff and alcian blue staining than did tubular carcinomas. These findings suggest that C2-O-sLe(x) is a tumor-associated antigen that may play a role in the invasiveness and metastatic potential of certain types of canine gastric carcinomas.

[Introduction of the new DRG-based reimbursement system in German hospitals--a difficult operation? Experiences and possible solutions from the viewpoint of trauma surgery]. / Die Einführung des neuen DRG-basierten Entgeltsystems in Deutschen Krankenhäusern--Eine schwierige Operation? Erfahrungen und Lösungsansätze aus der Sicht der Unfallchirurgie.

Until recently the costs of patients in German hospitals have mainly been calculated according to the length of hospital stay. However, in December 1999, a dramatic change was announced in the social laws in the "Gesundheitsreform 2000." Beginning in 2003 a prospective payment system based on the classification of the "Australian Refined DRGs (AR-DRG)" will be introduced. DRGs are already used in quite a lot of industrialized countries and basically are "per case" payment systems that group patients with homogeneous average costs based on the diagnoses and procedures performed in the hospital. When preparing for this new system, the clinician has a lot of additional tasks. Besides correctly documenting all clinical findings with the ICD-10 Diagnoses and the German OPS301 procedure codes, a knowledge of the economical impact of the clinical decision is absolutely crucial. The most important task is the optimizing of all clinical treatment processes (e.g. by the introduction of clinical pathways), because only hospitals that can do highly efficient treatment will be able to survive in the upcoming competitive situation. In the Krankenhaus München--Schwabing DRGs have been used as a benchmarking tool since 1997. Based on valuable experience, many direct measures to optimize efficiency have been taken. Especially in patients with multiple trauma, it became evident that the use of efficient and standardized treatment can bring economic gain without loss of quality.

Multiple lesion transluminal coronary angioplasty in single and multivessel coronary artery disease: acute outcome and long-term effect.

Multiple lesion transluminal coronary angioplasty was performed in 428 patients. Angioplasty was attempted in 1,047 lesions (2.4/patient), with an angiographic success achieved in 94%: 2 lesions were attempted in 74%, 3 in 21%, 4 in 5% and 5 or more in 1% of cases. A clinical success was achieved in 404 (94%) of the patients: 95% with and 93% without prior surgery and in 94% of those with single vessel disease and 94% of those with multivessel disease. Significant complications occurred in 17 patients (4.0%): 11 (2.5%) had a transmural infarction, 9 (2.1%) required urgent surgery and 6 (1.4%) died. An apparent lesion recurrence occurred in 106 (26%) of 404 patients with 81 of 89 patients (91%) having a successful second angioplasty. A second apparent lesion recurrence occurred in 15 patients (19%), with 13 of the 15 patients having a successful third angioplasty. A sustained clinical improvement (mean follow-up period 28.3 +/- 16 months) was obtained in 208 (83%) of 250 patients with successful angioplasty. The cumulative probability of survival at 51 months was 93% in these 250 patients. Survival was adversely affected by the presence of prior bypass surgery (no prior surgery 97% versus prior surgery 81%; p less than 0.05). These data suggest that multiple lesion angioplasty can be successfully performed with a good success rate, an acceptable incidence of complications and a reasonable expectation of satisfactory long-term clinical improvement.

Percutaneous transluminal valvuloplasty in calcific aortic stenosis: the double balloon technique.

The double balloon technique for aortic valvuloplasty using a combined brachial and femoral artery approach with readily available angioplasty equipment is described. Online hemodynamic assessment of the patient during the procedure was provided by continuous, reflective spectorphotometric monitoring of the pulmonary artery mixed venous oxygen saturation and the arterial pressure in the femoral sheath. Successful valvuloplasty was achieved in 10 patients with the transaortic peak-to-peak systolic gradient reduced from 77.6 +/- 28 to 27.8 +/- 15 mmHg (P less than 0.001), the mean gradient from 62.0 +/- 24 to 26.0 +/- 15 mmHg (P less than 0.0007), and the aortic valve area increased from 0.56 +/- 0.3 to 1.03 +/- 0.6 cm2 (P less than 0.05). No significant complications were encountered. The double balloon technique for aortic valvuloplasty is an acceptable alternative method to the single balloon technique.

Percutaneous transluminal coronary angioplasty in patients over the age of 70 years.

Percutaneous transluminal coronary angioplasty (PTCA) was attempted in 109 patients over the age of 70 years (mean age 75.9). The patients' clinical characteristics showed 55% with multivessel coronary disease, 86% with significant angina pectoris (Class II to IV), 16% with prior bypass surgery, and 2.7% with left ventricular dysfunction. Angioplasty data showed the mean percent diameter stenosis was reduced from 83 +/- 12% to 15 +/- 16%. One lesion was dilated in 67%, two lesions in 30%, and three lesions in 3% of the patients. A successful dilatation was achieved in 138/148 lesions (89%) with 90/109 patients (83%) clinically improved at hospital discharge. Significant complications were encountered in six patients (5.5%): three transmural infarctions (2.8%), two mortalities (1.8%), and one emergency surgical procedure (0.9%). Follow-up data (greater than or equal to 1 year) are available in 77 patients: 68 patients (92%) had an improved anginal status [49 patients (65%) had no angina]. During a mean follow-up of 23 +/- 10 months, there were two myocardial infarctions and three deaths. A clinically apparent recurrence occurred in 14 patients (17%). Repeat angioplasty was successfully performed in 13/14 patients. Bypass surgery was performed during the follow-up in three patients (two of whom had had a recurrence after a second successful angioplasty; one patient died during elective surgery). Transluminal coronary angioplasty can be performed in patients over age 70 with a good success rate, an acceptable complication rate, a relatively low clinically apparent recurrence rate, and should be considered as a therapeutic modality and alternative for the selected geriatric patient.

Coronary angioplasty in patients who undergo multiple vessel angioplasties, and in patients who have multivessel coronary disease.

Percutaneous transluminal coronary angioplasty (PTCA) was performed in patients who underwent multiple vessel angioplasties (MVA) and in patients with multivessel coronary disease (MCD). There were 235 MVA patients who underwent 255 procedures during which 600 lesions were dilated. Two lesions were dilated in 75% of cases, and three lesions in 16% of cases. 93% of the lesions were successfully dilated and 91% of the patients were clinically improved. Complications encountered included a 1.6% mortality, a 3.5% incidence of transmural infarction, and a 2.4% emergency surgery rate. Significant complications occurred in 13 patients (4.6%). A clinically apparent restenosis occurred in 51 patients (24%) with a repeat PTCA performed in 25/26 patients (96%), elective surgery in 14 patients, and the remainder were treated medically. 82% of patients remain clinically improved more than one year after a successful PTCA. There were 331 MCD patients who underwent 387 procedures during which 662 lesions were dilated. One lesion was dilated in 47% of cases, two lesions in 39% of cases, three lesions in 9% of cases, and four or more lesions in 5% of cases. 91% of the lesions were successfully dilated, and 90% of the patients were clinically improved. Complications encountered included a 1.6% mortality, a 3.1% incidence of transmural infarction, and a 3.9% emergency surgery rate. Significant complications were encountered in 35 patients (9%). A clinically apparent restenosis occurred in 46 patients (21%) with a repeat PTCA performed in 29/31 patients (94%), elective surgery in nine patients, and the remainder were treated medically. 86% of patients remain clinically improved more than one year after a successful PTCA. PTCA can be performed in carefully selected patients undergoing MVA, and in MCD patients with a good success rate, a low complication rate, and a satisfactory long term benefit. However, a large clinical trial would be most beneficial in establishing PTCA's role in the treatment of extensive coronary disease.

Complex coronary angioplasty in patients with prior coronary artery bypass surgery, in situations utilizing multiple coronary angioplasties, and in coronary occlusions.

Coronary artery bypass surgery not only provides symptomatic relief but also may prolong life in a significant percentage of patients. Repeat bypass surgery succeeds in providing symptomatic relief in a lesser number of patients, but it is associated with a higher morbidity and mortality than primary operations. Angioplasty, an interventional, therapeutic catheter technique, is applicable to a large number of patients who have undergone bypass surgery and who are sufficiently symptomatic to require consideration of another revascularization procedure. The gratifying results of successful lesion dilatation coupled with clinical improvement of the patient, the acceptable mortality and morbidity statistics, and the long-term symptomatic relief are comparable to those for repeat coronary bypass graft surgery. In addition, technologic advances in angioplasty equipment, as well as more knowledgeable interventionists, will enable more lesions to be successfully reached, traversed, and dilated with, it is hoped, a lower morbidity and mortality. We would estimate that 30 to 50 per cent of those patients requiring repeat revascularization operations today can undergo an angioplasty procedure with at least comparable clinical results and better morbidity and mortality statistics than those achieved with repeat bypass surgery. Selected patients underwent transluminal coronary angioplasty of varying combinations of arterial and/or vein graft stenoses. A multiple dilatation procedure was defined as successful when dilatation was achieved in all lesions in which it was attempted or when the considered-critical stenosis was dilated successfully and the patient was clinically improved. Angioplasty was successful in 93 per cent of all lesions in which it was attempted, and these successful dilatations produced a clinical improvement in 92 per cent of the patients. No complication whatsoever was experienced in 81 per cent of cases. The complications encountered included a 1.3 per cent mortality rate, a myocardial infarction rate of 6.9 per cent (3.0 per cent per lesion attempted), and an emergency surgery rate of 2.6 per cent (1.1 per cent per lesion attempted). Follow-up data show that a sustained clinical improvement was obtained in 96.8 per cent of the patients in whom the procedure was successful (with or without a repeat angioplasty). These data indicate that multiple coronary angioplasties can be performed during the same procedure, that clinical improvement can be achieved, and that the complication rate is acceptable.(ABSTRACT TRUNCATED AT 400 WORDS)

In-hospital mortality rate in the National Heart, Lung, and Blood Institute Percutaneous Transluminal Coronary Angioplasty Registry.

Twenty-nine patients died among the first 3,079 patients enrolled in the NHLBI PTCA Registry. The overall morality rate was 0.9%; the mortality rate was 0.8% in patients with 1-vessel CAD, 1.0% in those with multivessel CAD (excluding left main CAD), and 3.8% in those with left main CAD (p less than 0.01). The in-hospital morality rate was significantly higher among women (p less than 0.01), in patients older than 60 years, in patients with previous CABG (p less than 0.01), the presence of left main CAD, (p less than 0.01), in patients who required dilatation of a vein graft stenosis (p less than 0.05), and in patients who had had angina for longer than 6 months (p less than 0.01).

Percutaneous transluminal coronary angioplasty in patients with prior coronary artery bypass grafting.

Percutaneous transluminal coronary angioplasty (PTCA) has been used to treat patients with prior coronary artery bypass grafting who have stenosis of a saphenous vein graft and/or a native artery. During 53 months, 61 patients underwent 105 angioplasty attempts. Eighty lesions (76%) were successfully dilated in 46 of 61 patients (75%). Success in a patient was determined by a greater than or equal to 20% decrease in the percent diameter stenoses coupled with an improved clinical response; 52 patients had one prior CABG and nine patients had two or more prior CABGs. Multivessel disease was present in 56 patients (92%). A vein graft stenosis was successfully dilated in 26 of 33 cases (79%)--19 of 25 (76%) at an anastomotic site and seven of eight (88%) in the graft body. An arterial stenosis was successfully dilated in 37 of 52 cases (71%)--18 of 22 (82%) in the left anterior descending, 13 of 22 (59%) in the circumflex, 21 of 26 (81%) in the right coronary, and two (100%) in the left main coronary artery. There is no statistically significant difference in the incidence of success in dilating a vein graft or native artery. Complications included: one emergency CABG (1.6%), three myocardial infarctions (4.9%), and two deaths (3.3%). There were 15 unsuccessful PTCAs: Ten patients had elective CABG, one had emergency CABG, two received medical treatment, and two died. Forty-six patients are being followed-up: Twenty-eight (61%) continue to do clinically well, seven (15%) had another PTCA and remain well, and 10 (16%) had elective CABG because of restenosis and/or disease progression. There was one late death and one late myocardial infarction. Thus, 35 patients (57%) had continued clinical success without the need for repeat CABG; 89% had no angina or improved angina, and 90% had improved exercise treadmill results. PTCA is technically feasible in selected patients with prior CABG and can achieve a clinical response with an acceptable complication rate when compared to repeat CABG.

Employment and recreation patterns in patients treated by percutaneous transluminal coronary angioplasty: a multicenter study.

Employment and recreational patterns were analyzed in 279 patients who underwent percutaneous transluminal coronary angioplasty (PTCA) for treatment of symptomatic coronary artery disease. PTCA was successful in 180 patients (65%). When it was unsuccessful, coronary artery bypass graft surgery was usually performed (80%). Return-to-work rates were high irrespective of the outcome of PTCA. Of patients employed full-time or part-time before treatment, 98.5% of those who had successful PTCA alone and 97% of those whose PTCA was unsuccessful but who underwent uncomplicated coronary artery bypass surgery maintained or improved their work status. In a subgroup of men who had been employed in occupations requiring physical labor, 85% of the men whose PTCA was successful returned to work, compared with 68% of those whose PTCA was unsuccessful. The interval from attempted PTCA to return to work was significantly shorter in the successfully treated group; in patients with successful PTCA, the median time to return to work was 14 days, compared with 60 days in patients in whom PTCA was unsuccessful (p less than 0.001). During follow-up, patients with successful PTCA had less angina and were more active in recreational activities than patients who required alternative treatments.

Psychosocial and medical effects of vasectomy in a sample of health plan subscribers.

The effects of vasectomy on psychosocial adjustment assessed by interviews, and on physical health assessed by physicians and other medical personnel, were examined by comparing 33 vasectomized men with a matched group of 33 non-vasectomized men in a pre-paid health plan. Consistent with favorable self-reports in earlier studies, the vasectomized men experienced no greater marital, job, or general living stress than the non-vasectomized men. In fact the psychosocial adjustment of the vasectomized men appeared superior to that of the non-vasectomized men, perhaps because of reduced anxiety about unwanted pregnancy. The hypothesis advanced by some investigators that vasectomized men exaggerate their masculinity in an overcompensating maneuver to reduce a perceived threat to their masculinity was not supported; instead, men volunteering for vasectomy exhibited as many masculine traits prior to the operation as after it. There was no evidence that vasectomy led to either impairment or enhancement of medical health.

PIP: 33 vasectomized men were compared with 33 nonvasectomized men who were members of the health plan at the Kaiser Foundation Hospital, Portland, Oregon. The 1st study used data from a structured interview to examine psychosocial adjustment, masculinity, and rigidity. Little difference was found between the 2 groups except that vasectomized men reporter greater marital happiness and better health and appetite. This may be due to lack of worry about pregnancy. Health plan records were examined for illness as part of the 2nd study. Both groups had similar treatment records and vasectomized men did not have more ailments with strong psychological components than nonvasectomized.