Expression profiling in progressive stages of fumarate-hydratase deficiency: the contribution of metabolic changes to tumorigenesis

Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is caused by mutations in the Krebs cycle enzyme fumarate hydratase (FH). It has been proposed that "pseudohypoxic" stabilization of hypoxia-inducible factor-α (HIF-α) by fumarate accumulation contributes to tumorigenesis in HLRCC. We hypothesized that an additional direct consequence of FH deficiency is the establishment of a biosynthetic milieu. To investigate this hypothesis, we isolated primary mouse embryonic fibroblast (MEF) lines from Fh1-deficient mice. As predicted, these MEFs upregulated Hif-1α and HIF target genes directly as a result of FH deficiency. In addition, detailed metabolic assessment of these MEFs confirmed their dependence on glycolysis, and an elevated rate of lactate efflux, associated with the upregulation of glycolytic enzymes known to be associated with tumorigenesis. Correspondingly, Fh1-deficient benign murine renal cysts and an advanced human HLRCC-related renal cell carcinoma manifested a prominent and progressive increase in the expression of HIF-α target genes and in genes known to be relevant to tumorigenesis and metastasis. In accord with our hypothesis, in a variety of different FH-deficient tissues, including a novel murine model of Fh1-deficient smooth muscle, we show a striking and progressive upregulation of a tumorigenic metabolic profile, as manifested by increased PKM2 and LDHA protein. Based on the models assessed herein, we infer that that FH deficiency compels cells to adopt an early, reversible, and progressive protumorigenic metabolic milieu that is reminiscent of that driving the Warburg effect. Targets identified in these novel and diverse FH-deficient models represent excellent potential candidates for further mechanistic investigation and therapeutic metabolic manipulation in tumors.     

Using hospital administrative data to evaluate the knowledge-to-action gap in pressure ulcer preventive care

Rationale, aims and objectives  Issues of overuse, underuse and misuse are paramount and lead to avoidable morbidity and mortality. Although evidence-based practice is advocated, the widespread implementation of this kind of practice remains a challenge. This is also the case for evidence-based practice related to the prevention of pressure ulcers, which varies widely in process and outcome in Belgian hospital care. One major obstacle to bridging this knowledge-to-action gap is data availability. We propose using large-scale hospital administrative data combined with the latest evidence-based methods as part of the solution to this problem.
Method  To test our proposal, we applied this approach to pressure ulcer prevention, using an administrative dataset with regard to 6030 patients in 22 Belgian hospitals as a sample of nationally available data. Methods include a systematic review approach, evidence grading, recommendations formulation, algorithm construction, programming of the rule set and application on the database.
Results  We found that Belgian hospitals frequently failed to provide appropriate prevention care. Significant levels of underuse, up to 28.4% in pressure ulcer prevention education and 17.5% in the use of dynamic systems mattresses, were detected. Figures for overuse were mostly not significant. Misuse couldn't be assessed.
Conclusions  These results demonstrate that this approach can indeed be successfully used to bridge the knowledge-to-action gap in medical practice, by implementing an innovative method to assess underuse and overuse in hospital care. The integrative use of administrative data and clinical applications should be replicated in other patient groups, other datasets and other countries.     

Effects of fatty and lean fish intake on blood pressure in subjects with coronary heart disease using multiple medications

BACKGROUND: Intake of fish and long-chain n-3 fatty acids has been of wide interest due to their beneficial effects on cardiovascular risk factors and lower coronary heart disease (CHD) risk. AIM OF THE STUDY: The aim of this pilot study was to examine the effects of fatty fish and lean (white) fish on fatty acid composition of serum lipids and cardiovascular risk factors in subjects with CHD using multiple drugs for this condition. METHODS: The study was an 8-week controlled, parallel intervention. Inclusion criteria were myocardial infarction or unstable ischemic attack, age under 70 years, use of betablockers and presence of sinus rhythm. The subjects were randomized to one of the following groups: 4 meals/week fatty fish (n = 11), 4 meals/week lean fish (n = 12) and control diet including lean meat (n = 10). RESULTS: The mean (+/-SD) of reported fish meals per week was 4.3 +/- 0.4, 4.7 +/- 1.1 and 0.6 +/- 0.4 in the groups, respectively. The proportions of eicosapentaenoic and docosahexaenoic acids in serum lipids increased in the fatty fish group only (P < 0.05). Systolic and diastolic blood pressure levels decreased in the lean fish group (0 vs. 8 week: 3.5 +/- 3.2 and 4.6 +/- 3.6%, respectively, P < 0.05). Serum total triglyceride concentration did not significantly change. HDL cholesterol concentration change differed among groups but without significant post hoc differences. Apolipoprotein A-1 concentration decreased in the control group (0 vs. 8 week, P < 0.05). Coagulation factors, 25-hydroxy vitamin D, and heart rate variability (24 h Holter) did not change among the groups. CONCLUSIONS: Our results suggest that intake of lean fish at least four times per week could reduce blood pressure levels in CHD patients.     

Quantitative Evaluation of Spontaneously and Surgically Repaired Rabbit Articular Cartilage Using Intra-Articular Ultrasound Method in situ

During the last decade, a major effort has been devoted to developing surgical methods for repairing localized articular cartilage lesions. Despite some promising results no ultimate breakthrough in surgical cartilage repair has been achieved. Improvements in repair techniques would benefit from more sensitive and quantitative methods for long-term follow-up of cartilage healing. In this study, the potential of a new ultrasound technique for detecting the compositional and structural changes in articular cartilage after surgery, using recombinant human type II collagen gel and spontaneous repair was, investigated. Rabbit knee joints containing intact (n = 13) and surgically (n = 8) or spontaneously (n = 5) repaired tissue were imaged in situ at 6 months after the operation using a clinical intravascular high-frequency (40 MHz) ultrasound device. Based on the ultrasound raw data, ultrasound reflection coefficient (R), integrated ultrasound reflection coefficient (IRC), apparent integrated backscattering coefficient (AIB) and ultrasound roughness index (URI) were determined for each sample. URI was significantly higher in both repair groups than in intact cartilage (p < 0.05). The reflection parameters (R and IRC) were significantly lower in surgically repaired cartilage (p < 0.05) than in intact cartilage. Furthermore, AIB was significantly higher in surgically repaired cartilage than in intact tissue (p < 0.05). To conclude, the integrity of the rabbit articular cartilage repair could be quantitatively evaluated with the nondestructive ultrasound approach. In addition, clinically valuable qualitative information on the changes in cartilage integration, structure and composition could be extracted from the ultrasound images. In the present study, the structure and properties of repaired tissue were inferior to native tissue at 6 months after the operation. The applied ultrasound device and probes are FDA approved and, thus, applicable for the quantitative in vivo evaluation of human articular cartilage. (E-mail: tviren@hytti.uku.fi)     

Local aldosterone synthesis in the large intestine of mouse: An ex vivo incubation study

ObjectiveTo investigate the regulation of local aldosterone synthesis by physiological stimulants in the murine gut.MethodsMale mice were fed for 14 days with normal, high (1.6%) or low (0.01%) sodium diets. Tissue liver receptor homolog-1 and aldosterone in the colon and caecum were detected using an enzyme-linked immunosorbent assay (ELISA). Released corticosterone and aldosterone in tissue incubation experiments after stimulation with angiotensin II (Ang II) and dibutyryl-cAMP (DBA; the second messenger of adrenocorticotropic hormone) were assayed using an ELISA. Tissue aldosterone synthase (CYP11B2) protein levels were measured using an ELISA and Western blots.ResultsIn incubated colon tissues, aldosterone synthase levels were increased by a low-sodium diet; and by Ang II and DBA in the normal diet group. Release of aldosterone into the incubation buffer was increased from the colon by a low-sodium diet and decreased by a high-sodium diet in parallel with changes in aldosterone synthase levels. In mice fed a normal diet, colon incubation with both Ang II and DBA increased the release of aldosterone as well as its precursor corticosterone.ConclusionLocal aldosterone synthesis in the large intestine is stimulated by a low-sodium diet, dibutyryl-cAMP and Ang II similar to the adrenal glands.     

Formation of dihydrate from carbamazepine anhydrate in aqueous conditions

According to our observations, carbamazepine dihydrate crystals grow by the whisker mechanism (or mechanisms). The habit, the dimensions of the crystals and their rapid growth indicate this. Also the strong orientation in the X-ray diffractogram is in harmony with the whisker theory. The whisker growth explains also the rapid transition from the anhydrous form to the dihydrate one in water.

Dihydrate whiskers which are loosely in the solution as well as ones fastened to the surface of the anhydrous mother crystal has been observed. The oriented crystal structure of carbamazepine dihydrate may be at least one important factor in the growth mechanism of carbamazepine dihydrate whiskers.     

Sublethal damage repair after fractionated irradiation in endometrial cancer cell lines tested with the 96-well plate clonogenic assay

Two long-established and seven newly established endometrial adenocarcinoma cell lines were tested for their capacity to repair sublethal damage after fractionated irradiation. Cell survival was determined with the 96-well plate clonogenic assay based on limiting dilutions. Total radiation doses of 0.75 Gy, 1.25 Gy, 2.50 Gy, 5.00 Gy and 7.50 Gy were used either as a single dose or divided into two or three equal fractions with a 24 h interval. Survival data were fitted to the linear quadratic model, and the area under the survival curve (AUC), equivalent to the mean inactivation dose, was obtained with numerical integration. The amount of sublethal damage repair (SLDR) was expressed as an area-under-the-curve (AUC) ratio comparing survivals from fractionated-dose with those from single-dose experiments. SLDR capacity of the cell lines expressed as an AUC ratio varied between 1.00 and 1.59, and the mean was 1.17. Two highly radiosensitive cell lines were found to be SLDR-deficient, but most of the cell lines studied had some SLDR capacity. We have earlier shown that endometrial cancer cell lines as a group are more radiosensitive than squamous-cell carcinoma (SCC) lines. Data obtained in this study suggest that the capacity for SLDR in these cell lines is rather limited compared with the majority of SCC lines tested. This finding underlines further the high radioresponsiveness of endometrial cancer.Abbreviations SLDR sublethal damage repair - SCC squamous-cell carcinoma - AUC area under the curve This study was financially supported by the Southwestern Division of the Finnish Cancer Society and the Turku University FoundationPresented at the Fourth Biennial Meeting of the International Gynecologic Cancer Society, Aug 29-Sep 2 1993, in, Stockholm, Sweden     

Multi‐parametric MRI characterization of enzymatically degraded articular cartilage

Several laboratory and rotating frame quantitative MRI parameters were evaluated and compared for detection of changes in articular cartilage following selective enzymatic digestion. Bovine osteochondral specimens were subjected to 44 h incubation in control medium or in collagenase or chondroitinase ABC to induce superficial collagen or proteoglycan (glycosaminoglycan) alterations. The samples were scanned at 9.4 T for T₁, T_{1 Gd} (dGEMRIC), T₂, adiabatic T_{1 ρ}, adiabatic T_{2 ρ}, continuous‐wave T_{1 ρ}, T_RAFF2, and T_{1 sat} relaxation times and for magnetization transfer ratio (MTR). For reference, glycosaminoglycan content, collagen fibril orientation and biomechanical properties were determined. Changes primarily in the superficial cartilage were noted after enzymatic degradation. Most of the studied parameters were sensitive to the destruction of collagen network, whereas glycosaminoglycan depletion was detected only by native T₁ and T_{1 Gd} relaxation time constants throughout the tissue and by MTR superficially. T₁, adiabatic T_{1 ρ}, adiabatic T_{2 ρ}, continuous‐wave T_{1 ρ}, and T_{1 sat} correlated significantly with the biomechanical properties while T_{1 Gd} correlated with glycosaminoglycan staining. The findings indicated that most of the studied MRI parameters were sensitive to both glycosaminoglycan content and collagen network integrity, with changes due to enzymatic treatment detected primarily in the superficial tissue. Strong correlation of T₁, adiabatic T_1ρ, adiabatic T_{2 ρ}, continuous‐wave T_{1 ρ}, and T_{1 sat} with the altered biomechanical properties, reflects that these parameters were sensitive to critical functional properties of cartilage. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1111–1120, 2016.     

Measuring cost-effectiveness of secondary health care: feasibility and potential utilization of results

OBJECTIVES: Whether cost-effectiveness of secondary health care can be measured in a simple, yet commensurate way was studied. METHODS: Approximately 4,900 patients' health-related quality of life scores before and after treatment were measured. Used were a combination of quality of life data with diagnostic and financial indicators routinely collected in the hospital. RESULTS: Seventy percent of patients returned the first questionnaire and the informed written consent to participate. Of these patients, 80 percent also returned the second questionnaire sent out 3 to 12 months after treatment, depending on clinical specialty and diagnostic category. The routine of sending out questionnaires could be automated in such a way that data collection required only a limited amount of extra work. Patients were generally satisfied with the fact that the hospital was interested in their well-being also after treatment. No physician offered the chance to participate refused data collection in the patient group he or she was responsible for. The attitudes of the nursing staff were generally positive toward data collection, although it caused some extra work for some of them. The possibility of relating already routinely collected financial performance indicators with a relevant measure of treatment effectiveness, opened prospects for refined analysis of cost-effectiveness of secondary health care. CONCLUSIONS: Routine collection of health-related quality of life data as an indicator of treatment effectiveness is feasible, requires only a small amount of extra work, and is potentially very useful when combined with existing measures of hospital performance.     

Individual quality of life following radical prostatectomy in men with prostate cancer

PURPOSE: The aim of this study was to examine the individual quality of life (QoL) of men following radical prostatectomy for prostate cancer. The following research questions were addressed: (a) What are the most important areas of quality of life for men following radical prostatectomy? (b) How do these men rate their satisfaction in each area and what is the relative importance of each area to their overall quality of life? METHODS: The purposive sample consisted of 11 men with prostate cancer who had undergone a radical prostatectomy 3 to 4 months earlier. QoL was examined using the SEIQoL-DW (Schedule for the Evaluation of Individual QoL: A Direct Weighting Procedure). The data were analyzed by means of qualitative content analysis (five most important QoL areas). FINDINGS: The 11 respondents named a total of 55 QoL areas which they described and labelled. They then rated their current satisfaction in each area, and how important each one was to them. A second analysis of the content was made to identify the main QoL areas. The 55 quality of life areas mentioned by respondents were reduced to the following categories: health, activity, family, relationship with a partner, autonomy, independence, hobby, financial security, and sexuality. Health, family, and relationship with a partner are the thee areas which had the most impact on QoL. Overall, the respondents had a high quality of life value. Impotence and incontinence did not appear to have a very negative impact on quality of life. CONCLUSIONS: SEIQoL-DW was used for the first time in patients with prostate cancer. In a urology department where nurses and patients are confronted daily with the topics of intimacy, sexuality, and sense of embarrassment, more importance should be placed on the topic of sexuality when taking a patient history. Nurses should be trained in communication techniques that enable them to engage patients in a safe and therapeutic dialogue about their sexual concerns related to the diagnosis of prostate cancer. SEIQoL-DW can support the communication with patients.