Analytical sensitivity and stability of DNA methylation testing in stool samples for colorectal cancer detection

Background

Stool-based molecular tests hold large potential for improving colorectal cancer screening. Here, we investigated the analytical sensitivity of a DNA methylation assay on partial stool samples, and estimated the DNA degradation in stool over time. In addition, we explored the detection of DNA methylation in fecal immunochemical test (FIT) fluid.

Materials and Methods

Partial stool samples of colonoscopy-negative individuals were homogenized with stool homogenization buffer, spiked with different numbers of HCT116 colon cancer cells and kept at room temperature for 0, 24, 48, 72 and 144?h before DNA isolation. Analytical sensitivity was determined by the lowest number of cells that yielded positive test results by DNA methylation or mutation analysis. DNA methylation in FIT fluid was measured in 11 CRC patients and 20 control subjects.

Results

The analytical sensitivity for detecting DNA methylation was 3000 cells per gram stool, compared to 60000 cells per gram stool for detection of DNA mutations in the same stool samples. No degradation up to 72?h was noted when a conservation buffer was used. DNA methylation was detected in 4/11 CRC FIT samples and in none of the 20 control FIT samples.

Conclusions

Methylation based stool DNA testing showed a high analytical sensitivity for tumor DNA in partial stool samples, which was hardly influenced by DNA degradation over time, provided an adequate buffer was used. The feasibility of detecting DNA methylation in FIT fluid demonstrates the opportunity to combine testing for occult blood with DNA methylation in the same collection device.     

Intrapartum Rupture of the Uterus Diagnosed by Ultrasound

Background

Spontaneous uterine rupture is a life threatening obstetrical emergency encountered infrequently in the emergency department. Its diagnosis is often missed or delayed, leading to maternal and fetal mortality.

Method

We present a case of ruptured uterus diagnosed by ultrasound in a 33-year-old gravid female with two previous cesarean sections.

Objective

To show the role of ultrasound in uterine rupture

Result

Ultrasound demonstrates uterine laceration and intra-abdominal dislocation of placenta and foetus

Conclusion

Because of the severity of the complication and the great variation of symptoms in connection with it, the authors encourage the use of ultrasound screening in the detection of this rare, but often catastrophic complication.     

Neurolytic Blocks of the Sympathetic Axis for the Treatment of Visceral Pain in Cancer

Pain due to cancer is frequently visceral, and neurolysis of the sympathetic axis has been shown to be an effective and safe method for treating this visceral pain. Several studies have documented the efficacy of neurolytic blocks both by a reduction in the intensity of pain and by a decrease in opioid consumption. Neurolysis of the sympathetic axis should be incorporated into the pain specialist's arsenal as an adjuvant to oral pharmacologic therapy.     

A prospective,randomized, pragmatic,health outcomes trial evaluating the incorporation of hylan G-F 20 into the treatment paradigm for patients with knee osteoarthritis (Part 2 of 2): economic results

OBJECTIVE: Viscosupplementation with hylan G-F 20 has recently become registered for treatment of patients with osteoarthritis (OA) of the knee in most parts of the world. The cost effectiveness and cost utility of this new therapeutic modality were determined as part of a Canadian prospective, randomized, 1-year, open-label, multicentered trial. DESIGN: A total of 255 patients were randomized to 'Appropriate care with hylan G-F 20' (AC+H) or 'Appropriate care without hylan G-F 20' (AC). Costs (1999 Canadian dollars) were collected from the societal viewpoint and included all costs related to OA of the knee and OA in all joints. Patients completed a number of outcomes questionnaires including the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the Health Utilities Index Mark 3 (HUI3). Data were collected at clinic visits (baseline, 12 months) and by telephone (1, 2, 4, 6, 8, 10, and 12 months). RESULTS: The AC+H group over the year had higher costs ($2125-$1415=$710, P< 0.05), more patients improved (69%-40%=29%,P =0.0001), greater increases in HUI3 (0.13-0.03=0.10, P< 0.0001) and increased quality-adjusted life years (QALYs) (0.071, P< 0.05). The incremental cost-effectiveness ratio was $2505/patient improved. The incremental cost-utility ratio was $10000/QALY gained. Sensitivity analyses and a second cost perspective gave similar results. CONCLUSION: The cost-utility ratio is below the suggested Canadian adoption threshold. The results provide strong evidence for adoption of treatment with hylan G-F 20 in the patients and settings studied in the trial.     

Oral infectious diseases: a potential risk factor for HIV virus recrudescence?

As the highly active antiretroviral therapy (HAART) has transitioned human immunodeficiency virus (HIV) infection into a 'chronic disease' management strategy, there is growing evidence that infection with non-HIV pathogens in HIV+ patients may have important public health implications in undermining HAART success and acquired immunodeficiency syndrome progression. Several bacterial and host cell products during infections with non-HIV pathogens have shown the capacity to regulate HIV replication in latently infected cells. A high prevalence of oral infections caused by bacteria, viruses and fungi has been described in HIV+ patients, including periodontal disease. The oral cavity appears to be a site of HIV pathogenesis and potential reservoir for the disease as HIV RNA and DNA forms are present in saliva as well as in gingival crevicular fluid, and oral epithelial cells are susceptible to either cell free or cell-associated HIV infection. The clinical and biological bases of potential associations between chronic oral inflammatory disorders, such as periodontal disease, and exacerbation of HIV viraemia have received little attention. This review attempts to evaluate the current understanding of HIV reactivation as a result of co-infection and/or inflammation induced by non-HIV pathogens in HIV-infected patients, and presents a hypothetic model about the potential role of periodontitis as a global oral infection that potentially contributes to HIV recrudescence.     

Heartbeat evoked potentials mirror altered body perception in depressed patients

Objective

Awareness of stimuli originating inside of the body (interoceptive awareness) is thought to have an impact on psychopathology. The aim of the present study was to analyze whether heartbeat perception accuracy is reduced in depressed patients. Furthermore, we investigated whether putative differences are reflected in heartbeat-evoked potentials.

Method

We assessed the heartbeat perception score in 16 depressed patients and in matched healthy controls. A 63-channel EEG was recorded while participants counted pseudo-randomly presented target tones or heartbeats during a fixed number of cardiac cycles. ECG R-waves served as the trigger for EEG averaging. The cardiac-field artifact was minimized using independent component analysis and current–source density.

Results

Behaviorally, the depressed sample showed less accurate heartbeat perception in comparison to the control group (p = .011). The two groups also demonstrated psychophysiological differences, showing that heartbeat-evoked potentials were significantly reduced in depressed patients.

Conclusions

Our results suggest that heartbeat evoked potentials are objective markers of altered bodily awareness. Reduced interoception during depression may be linked to alexithymia, as well as to both decreased capacity for decision-making and for cognitive processing.

Significance

It may be helpful to practice interoceptive awareness to improve depressive symptoms, for example by practicing meditation.     

Anterior cruciate ligament allograft transplantation for intraarticular ligamentous reconstruction

Summary A multiplicity of surgical operations have been developed in an attempt to achieve satisfactory function after anterior cruciate ligament (ACL) repair. None of these procedures have been able to reproduce the fiber organization anatomy of attachment site, vascularity, or function of the ACL. Twenty-nine foxhounds received a deep-frozen bone-ACL-bone allograft and a ligament augmentation device (LAD). Biomechanical, microvascular, and histological changes were evaluated 3, 6, and 12 months following implantation. The maximum loads of the allograft/LADs were 34.3% (387.2N) after 3 months, 49.3% (556.6N) after 6 months, and 61.1% (698.8N) after a year. The maximum load was 69.1% (780 N). In general, after 6 months the allografts showed normal collagen orientation. The allografts demonstrated no evidence of infection or immune reaction. No bone ingrowth into the LAD was observed. Polarized light microscopy and periodic acid-schiff staining showed that the new bone-ligament substance interface had intact fiber orientation at the area of the ligament insertion. Microvascular examination using the Spalteholtz technique revealed revascularization and the importance of an infrapatellar fat pad for the nourishment of ACL allografts.