Cathepsin B inhibition limits bone metastasis in breast cancer

Metastasis to bone is a major cause of morbidity in breast cancer patients, emphasizing the importance of identifying molecular drivers of bone metastasis for new therapeutic targets. The endogenous cysteine cathepsin inhibitor stefin A is a suppressor of breast cancer metastasis to bone that is coexpressed with cathepsin B in bone metastases. In this study, we used the immunocompetent 4T1.2 model of breast cancer which exhibits spontaneous bone metastasis to evaluate the function and therapeutic targeting potential of cathepsin B in this setting of advanced disease. Cathepsin B abundancy in the model mimicked human disease, both at the level of primary tumors and matched spinal metastases. RNA interference-mediated knockdown of cathepsin B in tumor cells reduced collagen I degradation in vitro and bone metastasis in vivo. Similarly, intraperitoneal administration of the highly selective cathepsin B inhibitor CA-074 reduced metastasis in tumor-bearing animals, a reduction that was not reproduced by the broad spectrum cysteine cathepsin inhibitor JPM-OEt. Notably, metastasis suppression by CA-074 was maintained in a late treatment setting, pointing to a role in metastatic outgrowth. Together, our findings established a prometastatic role for cathepsin B in distant metastasis and illustrated the therapeutic benefits of its selective inhibition in vivo.     

Diffusion‐weighted imaging of human carotid artery using 2D single‐shot interleaved multislice inner volume diffusion‐weighted echo planar imaging (2D ss‐IMIV‐DWEPI) at 3T: Diffusion measurement in atherosclerotic plaque

Purpose:

To determine if 2D single‐shot interleaved multislice inner volume diffusion‐weighted echo planar imaging (ss‐IMIV‐DWEPI) can be used to obtain quantitative diffusion measurements that can assist in the identification of plaque components in the cervical carotid artery.

Materials and Methods:

The 2D ss‐DWEPI sequence was combined with interleaved multislice inner volume region localization to obtain diffusion weighted images with 1 mm in‐plane resolution and 2 mm slice thickness. Eleven subjects, six of whom have carotid plaque, were studied with this technique. The apparent diffusion coefficient (ADC) images were calculated using DW images with b = 10 s/mm² and b = 300 s/mm².

Results:

The mean ADC measurement in normal vessel wall of the 11 subjects was 1.28 ± 0.09 × 10^?3 mm²/s. Six of the 11 subjects had carotid plaque and ADC measurements in plaque ranged from 0.29 to 0.87 × 10^?3 mm²/s. Of the 11 common carotid artery walls studied (33 images), at least partial visualization of the wall was obtained in all ADC images, more than 50% visualization in 82% (27/33 images), and full visualization in 18% (6/33 images).

Conclusion:

2D ss‐IMIV‐DWEPI can perform diffusion‐weighted carotid magnetic resonance imaging (MRI) in vivo with reasonably high spatial resolution (1 × 1 × 2 mm³). ADC values of the carotid wall and plaque are consistent with similar values obtained from ex vivo endarterectomy specimens. The spread in ADC values obtained from plaque indicate that this technique could form a basis for plaque component identification in conjunction with other MRI/MRA techniques. J. Magn. Reson. Imaging 2009;30:1068–1077. © 2009 Wiley‐Liss, Inc.

     

The Utility of Pudendal Nerve Terminal Motor Latencies in Idiopathic Incontinence

Purpose Pudendal nerve terminal motor latency testing has been used to test for pudendal neuropathy, but its value remains controversial. We sought to clarify the relationship of pudendal nerve terminal motor latency to sphincter pressure and level of continence in a cohort of patients with intact anal sphincters and normal pelvic floor anatomy. Methods We reviewed 1,404 consecutive patients who were evaluated at our pelvic floor laboratory for fecal incontinence. From this group, 83 patients had intact anal sphincters on ultrasound and did not have internal or external rectal prolapse during defecography. These patients were evaluated by pudendal nerve terminal motor latency testing, a standardized questionnaire, and anorectal manometry, which measured resting and squeeze anal pressures. Incontinence scores were calculated by using the American Medical Systems Fecal Incontinence Score. Values were compared by using the Fisher’s exact test and Wilcoxon’s rank-sum test; and significance was assigned at the P < 0.05 level. Results 1) Using a 2.2-ms threshold, 28 percent of patients had prolonged pudendal nerve terminal motor latency unilaterally and 12 percent bilaterally. 2) At a 2.4-ms threshold, 18 percent of patients had prolonged pudendal nerve terminal motor latency unilaterally and 8 percent bilaterally. 3) Bilaterally prolonged pudendal nerve terminal motor latency was significantly associated with decreased maximum mean resting pressure and increased Fecal Incontinence Score, but not decreased maximum mean squeeze pressure, at both 2.2-ms and 2.4-ms thresholds. 4) Unilaterally prolonged pudendal nerve terminal motor latency was not associated with maximum mean resting pressure, maximum mean squeeze pressure, or fecal incontinence score at either threshold. Conclusions The majority of incontinent patients with intact sphincters have normal pudendal nerve terminal motor latency. Bilaterally but not unilaterally prolonged pudendal nerve terminal motor latency is associated with poorer function and physiology in the incontinent patient with an intact sphincter. Presented at the meeting of The American Society of Colon and Rectal Surgeons, Dallas, Texas, May 8 to 13, 2004. Reprints are not available.     

A prospective study of real-time panfungal PCR for the early diagnosis of invasive fungal infection in haemato-oncology patients

A blinded prospective study was performed to determine whether screening of whole blood using a real-time, panfungal polymerase chain reaction (PCR) technique could predict the development of invasive fungal infection (IFI) in immunocompromised haemato-oncology patients. In all, 78 patients (125 treatment episodes) were screened twice weekly by real-time panfungal PCR using LightCyclertrade mark technology. IFI was documented in 19 treatment episodes (five proven, three probable and 11 possible), and in 12, PCR was sequentially positive. PCR positivity occurred in: 4/5 proven; 2/3 probable; 6/11 possible; and 29/106 with no IFI. In 8/12 with IFI and sequentially positive PCR results, PCR positivity occurred before (median 19.5 days) and in 4/12 (median 10.5 days) after the initiation of empirical antifungal therapy. Based on sequential positive results for proven/probable IFI sensitivity, specificity, positive predictive value and negative predictive value were 75, 70, 15 and 98%, respectively. Real-time panfungal PCR is a sensitive tool for the early diagnosis of IFI in immunocompromised haemato-oncology patients. It may be most useful as a screening method in high-risk patients, either to direct early pre-emptive antifungal therapy or to determine when empirical antifungal therapy can be withheld in patients with antibiotic--resistant neutropenic fever. However, these strategies require further assessment in comparative clinical trials.     

The effect of exercise on the total water space and red blood cell volume of the gracilis muscle of the dog

The effect of increasing intensities of exercise on the water space and the red blood cell volume of the dog gracilis muscle was determined with 125I-labelled iodoantipyrine (n = 8) and 99mTc labelled red blood cells (n = 4). A sodium iodide scintillation probe attached to a two channel multiscaler and printer recorded the 125I activity of the muscle as a measure of the water space and the 99mTc activity as a measure of the red cell space. An electro-magnetic flow probe recorded muscle blood flow: aortic pressure was measured with a catheter in the mid-abdominal aorta. At least 30 min after i.v. administration of iodoantipyrine and red blood cells, the obturator nerve was electrically stimulated to produce muscle contraction at 1 contraction per second for 5 min. Data were recorded prior to exercise, during the 5 min exercise period, and for 10 min after exercise. During the greatest exercise period there was an eightfold increase in muscle blood flow, but only a small increase in [125I]iodoantipyrine activity (2.7 +/- 0.8%, 95% confidence limits) and 99mTc red blood cell activity (3.2 +/- 2.1%). Both values promptly returned to baseline during recovery. These data show that there is a minimal change in the water space and blood volume of the exercising dog gracilis muscle.     

The Karyopherin proteins,Crm1 and Karyopherin β1, are overexpressed in cervical cancer and are critical for cancer cell survival and proliferation

The Karyopherin proteins are involved in nucleo‐cytoplasmic trafficking and are critical for protein and RNA subcellular localization. Recent studies suggest they are important in nuclear envelope component assembly, mitosis and replication. Since these are all critical cellular functions, alterations in the expression of the Karyopherins may have an impact on the biology of cancer cells. In this study, we examined the expression of the Karyopherins, Crm1, Karyopherin β1 (Kpnβ1) and Karyopherin α2 (Kpnα2), in cervical tissue and cell lines. The functional significance of these proteins to cancer cells was investigated using individual siRNAs to inhibit their expression. Microarrays, quantitative RT‐PCR and immunofluorescence revealed significantly higher expression of Crm1, Kpnβ1 and Kpnα2 in cervical cancer compared to normal tissue. Expression levels were similarly elevated in cervical cancer cell lines compared to normal cells, and in transformed epithelial and fibroblast cells. Inhibition of Crm1 and Kpnβ1 in cancer cells significantly reduced cell proliferation, while Kpnα2 inhibition had no effect. Noncancer cells were unaffected by the inhibition of Crm1 and Kpnβ1. The reduction in proliferation of cancer cells was associated with an increase in a subG1 population by cell cycle analysis and Caspase‐3/7 assays revealed increased apoptosis. Crm1 and Kpnβ1 siRNA‐induced apoptosis was accompanied by an increase in the levels of growth inhibitory proteins, p53, p27, p21 and p18. Our results demonstrate that Crm1, Kpnβ1 and Kpnα2 are overexpressed in cervical cancer and that inhibiting the expression of Crm1 and Kpnβ1, not Kpnα2, induces cancer cell death, making Crm1 and Kpnβ1 promising candidates as both biomarkers and potential anticancer therapeutic targets. © 2008 Wiley‐Liss, Inc.     

Disruption of canonical TGFβ-signaling in murine coronary progenitor cells by low level arsenic

Exposure to arsenic results in several types of cancers as well as heart disease. A major contributor to ischemic heart pathologies is coronary artery disease, however the influences by environmental arsenic in this disease process are not known. Similarly, the impact of toxicants on blood vessel formation and function during development has not been studied. During embryogenesis, the epicardium undergoes proliferation, migration, and differentiation into several cardiac cell types including smooth muscle cells which contribute to the coronary vessels. The TGFβ family of ligands and receptors is essential for developmental cardiac epithelial to mesenchymal transition (EMT) and differentiation into coronary smooth muscle cells. In this in vitro study, 18 hour exposure to 1.34 μM arsenite disrupted developmental EMT programming in murine epicardial cells causing a deficit in cardiac mesenchyme. The expression of EMT genes including TGFβ2, TGFβ receptor-3, Snail, and Has-2 are decreased in a dose-dependent manner following exposure to arsenite. TGFβ2 cell signaling is abrogated as detected by decreases in phosphorylated Smad2/3 when cells are exposed to 1.34 μM arsenite. There is also loss of nuclear accumulation pSmad due to arsenite exposure. These observations coincide with a decrease in vimentin positive mesenchymal cells invading three-dimensional collagen gels. However, arsenite does not block TGFβ2 mediated smooth muscle cell differentiation by epicardial cells. Overall these results show that arsenic exposure blocks developmental EMT gene programming in murine coronary progenitor cells by disrupting TGFβ2 signals and Smad activation, and that smooth muscle cell differentiation is refractory to this arsenic toxicity.     

The management of PSA failure after radical radiotherapy for localized prostate cancer.

An asymptomatic rising serum prostate-specific antigen (PSA) level is the most common form of failure after radical radiotherapy for localized prostate cancer, but there is no consensus as to how it should be managed. This review addresses the following three questions concerning men with PSA failure after radiotherapy: (i) what is the course of the disease without further intervention?; (ii) what is the role of radical treatment, such as salvage prostatectomy?; and (iii) should androgen deprivation be started immediately or should it be delayed until clinical progression occurs? An algorithm for the management of PSA failure after radical radiotherapy for localized prostate cancer is proposed.     

Psychological factors in people at ultra-high risk of psychosis: comparisons with non-patients and associations with symptoms

BACKGROUND: There have been recent advances in the ability to identify people at high risk of developing psychosis. This has led to interest in the possibility of preventing the development of psychosis and provides the opportunity to investigate psychological mechanisms that may confer vulnerability to psychosis. METHOD: Fifty-eight patients at ultra-high risk of developing a first episode of psychosis were compared with 56 non-patients matched for age and occupational status on measures of meta-cognition, schizotypal traits, dysfunctional attitudes and distress. RESULTS: Analyses of covariance revealed that people at high risk of developing psychosis scored higher on measures of cognitive vulnerability, including negative meta-cognitive beliefs, beliefs about rejection and criticism from others, and discrepancies in self-perception, schizotypal traits and general mental distress. Correlational analyses revealed that negative meta-cognitive beliefs, dysfunctional attitudes and beliefs about rejection and criticism from others were positively associated with several dimensions of symptomatology in at-risk mental states (ARMS) patients. CONCLUSIONS: Cognitive and personality factors appear to characterize people at high-risk of developing psychosis and are associated with their distressing experiences. The clinical implications of these findings are discussed.