CSAP localizes to polyglutamylated microtubules and promotes proper cilia function and zebrafish development

The diverse populations of microtubule polymers in cells are functionally distinguished by different posttranslational modifications, including polyglutamylation. Polyglutamylation is enriched on subsets of microtubules including those found in the centrioles, mitotic spindle, and cilia. However, whether this modification alters intrinsic microtubule dynamics or affects extrinsic associations with specific interacting partners remains to be determined. Here we identify the microtubule-binding protein centriole and spindle-associated protein (CSAP), which colocalizes with polyglutamylated tubulin to centrioles, spindle microtubules, and cilia in human tissue culture cells. Reducing tubulin polyglutamylation prevents CSAP localization to both spindle and cilia microtubules. In zebrafish, CSAP is required for normal brain development and proper left-right asymmetry, defects that are qualitatively similar to those reported previously for depletion of polyglutamylation-conjugating enzymes. We also find that CSAP is required for proper cilia beating. Our work supports a model in which polyglutamylation can target selected microtubule-associated proteins, such as CSAP, to microtubule subpopulations, providing specific functional capabilities to these populations.     

Experience with 125I brachytherapy as a radical treatment for prostate cancer at the Russian X-ray Radiology Research Center

Prostate cancer is one of the most common malignancies among males. 125I brachytherapy is a current, effective, comparatively safe, and easy-to-reproduce treatment. The Russian X-ray Radiology Research Center has implanted 125I microsources in patients with localized and locally advanced prostate cancer since 2003; 689 125I implantations were performed in the past year. Tumor-specific survival after brachytherapy did not differ greatly from that following radical prostatectomy. Thus, brachytherapy is a current high-tech treatment for prostate cancer. This therapy shows fewer adverse postradiation effects than radiation teletherapy.     

Cancer de la prostate : la maladie localisée

Localized prostate cancer is characterized by a tumor confined to the prostate gland at clinical evaluation. Since the onset of PSA screening, the detection of localized prostate cancer has increased. Prognosis factors are clinical stadification, PSA value, PSA doubling time, tumor volume related to needle biopsy pathologic findings (Gleason score, percentage biopsies involved). Treatment depends on tumor prognosis, symptoms and performance status of the patient. Localized prostate cancer can be treated by surgery (radical prostatectomy, high intensity focused ultrasound) or radiotherapy (conformational radiation therapy, brachytherapy). Active follow-up can be proposed to very low risk patients.     

Salvage cryosurgery of the prostate after radiation failure

Radiation therapy is one option for patients with localized prostate cancer. Despite advances in delivering radiation to the prostate gland with therapies such as brachytherapy and/or external beam radiation therapy, urologists will be faced with managing patients with rising prostate-specific antigen values and with positive biopsy results secondary to radiation-recurrent prostate cancer. If the cancer is detected early, salvage therapy can be initiated. Since salvage prostatectomy is associated with significant morbidity, patients are often left with the option of either watchful waiting or temporary palliation with hormone deprivation therapy, with its attendant toxicity. The introduction of third-generation cryotechnology using 17-gauge CryoNeedlestrade mark (Oncura, Inc., Plymouth Meeting, PA) and the recent modifications in the technique of salvage cryosurgery have enabled cryosurgeons to eradicate these tumors safely and with significantly decreased morbidity. Selection and management of patients, as well as the contemporary results of salvage cryosurgery, are discussed in this article.     

An investigation of the effects from a urethral warming system on temperature distributions during cryoablation treatment of the prostate: A phantom study

Introduction of urethral warmers to aid cryosurgery in the prostate has significantly reduced the incidence of urethral sloughing; however, the incidence rate still remains as high as 15%. Furthermore, urethral warmers have been associated with an increase of cancer recurrence rates. Here, we report results from our phantom-based investigation to determine the impact of a urethral warmer on temperature distributions around cryoneedles during cryosurgery. Cryoablation treatments were simulated in a tissue mimicking phantom containing a urethral warming catheter. Four different configurations of cryoneedles relative to urethral warming catheter were investigated. For each configuration, the freeze–thaw cycles were repeated with and without the urethral warming system activated. Temperature histories were recorded at various pre-arranged positions relative to the cryoneedles and urethral warming catheter. In all configurations, the urethral warming system was effective at maintaining sub-lethal temperatures at the simulated surface of the urethra. The warmer action, however, was additionally demonstrated to potentially negatively impact treatment lethality in the target zone by elevating minimal temperatures to sub-lethal levels. In all needle configurations, rates of freezing and thawing were not significantly affected by the use of the urethral warmer. The results indicate that the urethral warming system can protect urethral tissue during cryoablation therapy with cryoneedles placed as close as 5 mm to the surface of the urethra. Using a urethral warming system and placing multiple cryoneedles within 1 cm of each other delivers lethal cooling at least 5 mm from the urethral surface while sparing urethral tissue.     

Salvage I-125 brachytherapy for locally-recurrent prostate cancer after radiotherapy

PurposeRetrospective, single-institution analysis of clinical outcomes and treatment-related toxicity in patients treated with salvage I-125 low-dose rate (LDR) brachytherapy (BT) for locally-recurrent prostate cancer after radiotherapy.Materials and methodsBetween 2008 and 2018, 30 patients with biopsy-confirmed prostate cancer recurrence underwent salvage treatment with I-125 LDR-BT. Of these 30 patients, 14 were previously treated with primary external beam radiotherapy (EBRT; median dose, 73 Gy) and 16 with primary I-125 LDR-BT (145 Gy and 160 Gy in 14 and 2 cases, respectively). At seed implantation, the mean age was 75.8 years, with a median Gleason score of 7 and pre-salvage PSA of <10 ng/mL. Six patients received androgen deprivation therapy for six months after relapse diagnosis. The prescribed salvage I-125 BT dose to the gland was 120−130 Gy, with dose restrictions of Dmax <135% (urethra) and <100% (rectum). Toxicity was evaluated according to the CTCAE scale (v4.0).ResultsAt a median follow-up of 45 months, the biochemical recurrence-free survival rates at 1, 3 and 5 years were 86.7%, 56.7% and 53.3%, respectively. Overall survival at 5 years was 87%. On the multivariate analysis, two variables were significant predictors of recurrence: PSA at relapse and nadir PSA post-salvage. Grade 3 genitourinary toxicity was observed in 5 patients (radiation-induced cystitis in 3 cases and urethral stenosis in 2) and G3 gastrointestinal toxicity in 3 patients (rectal bleeding).ConclusionSalvage therapy with I-125 brachytherapy is a safe and effective treatment option for locally-recurrent prostate cancer in previously-irradiated patients. High pre-salvage PSA and post-salvage nadir PSA values were significantly associated with a worse disease control after salvage I-125 LDR-BT. In well-selected patients, I-125 LDR-BT is comparable to other salvage therapies in terms of disease control and toxicity. However, more research is needed to determine the optimal management of locally-recurrent prostate cancer.     

Acute myeloid leukemia incidence following radiation therapy for localized or locally advanced prostate adenocarcinoma

Introduction: The effect of radiation therapy on acute myeloid leukemia incidence among prostate cancer patients has not been sufficiently elucidated despite evidence that acute myeloid leukemia is a consequence of therapeutic radiation in other primary malignancies. Therefore, we investigated the effect of definitive therapy with radiation therapy (external beam radiation therapy [EBRT] or brachytherapy) on acute myeloid leukemia incidence in a population-based cohort of patients with localized or locally advanced prostate cancer. Methods: We utilized the Surveillance, Epidemiology, and End Results database to identify a cohort of men (n = 168,612) with newly diagnosed prostate adenocarcinoma between January 1988 and December 2003. Cox proportional hazard regression was used to estimate the hazard ratios (HRs) and corresponding 95% confidence intervals (CIs) of acute myeloid leukemia incidence following definitive therapy with EBRT alone, brachytherapy alone, or surgery alone compared to no definitive therapy (i.e. no EBRT, brachytherapy, or surgery). Results: The cohort yielded 184 acute myeloid leukemia cases during 1,064,820 person-years of follow-up after prostate adenocarcinoma diagnosis. Patients treated with EBRT had a higher adjusted relative risk of developing acute myeloid leukemia than patients treated with brachytherapy or surgery when each therapy group was compared to patients who were not treated with definitive therapy (EBRT: HR = 2.05, 95% CI 1.29, 3.26; brachytherapy: HR = 1.22, 95% CI 0.46, 3.22; surgery: HR = 1.24, 95% CI 0.77, 1.98). Conclusions: Our findings suggest that acute myeloid leukemia incidence is a greater concern for patients treated with EBRT than brachytherapy for localized or locally advanced prostate adenocarcinoma.     

Basal cells of H-Dunning tumor are myoepithelial cells. A comparative immunohistochemical and ultrastructural study with male accessory sex glands and mammary gland

Recent immunohistochemical studies have shown that basal cells in human prostatic epithelium are not myoepithelial cells. Since in the literature the Dunning tumor, originally described as a rat prostate carcinoma derived from the dorsolateral prostate of a Copenhagen rat, was reported to have myoepithelial cells, a comparative immunohistochemical and ultrastructural study was performed in the H-, HIF- and AT3-lines of the Dunning tumor, the male accessory sex glands (ventral, dorsal, lateral prostate, coagulating gland, bulbourethral gland) and the mammary gland of both Copenhagen and Wistar rats. Mono- and polyclonal antibodies directed against intermediate filament proteins (cytokeratin, desmin, vimentin) and the contractile proteins (alpha-actin, muscle type specific myosin, tropomyosin) were used along with phalloidin decoration of F-actin. As in the human prostate, none of the rat prostate lobes in either strains did contain basal cells expressing cytokeratin along with alpha-actin, myosin and tropomyosin Cells representing fully differentiated myoepithelial cells, however, were present as anticipated in the mammary gland, the bulbourethral gland and the H-tumor line of the Dunning tumor. This finding is difficult to reconcile with the contention of a prostatic origin of the H-Dunning tumor. Further studies are required to classify the epithelial parental tissue in order to define the true origin of the H-Dunning tumor and the tumor lines derived thereof.     

Selenium level in benign and cancerous prostate

The dietary microelement selenium (Se) has been proposed as a potential chemopreventive agent for prostate cancer. This element is present in various amounts in all tissues. Little information is available on Se level in patients with prostate gland disorders. The levels of Se in prostatic gland of patients with prostate cancer, benign prostate hyperplasia, and healthy controls were examined. The Se level for benign prostate hyperplasia (156±30.6 ng/g) was the same as in the control group (157±26.0 ng/g), but in the gland of prostate cancer patients (182±34.1 ng/g wet weight), the Se level was significantly (p<0.01) higher than in both healthy controls and benign prostate hyperplasia. Thus, the Se level in human healthy controls is lower than in kidney and liver but higher compared with other tissues.     

Basal cells of H-Dunning tumor are myoepithelial cells

Summary Recent immunohistochemical studies have shown that basal cells in human prostatic epithelium are not myoepithelial cells. Since in the literature the Dunning tumor, originally described as a rat prostate carcinoma derived from the dorsolateral prostate of a Copenhagen rat, was reported to have myoepithelial cells, a comparative immunohistochemical and ultrastructural study was performed in the H-, HIF- and AT₃-lines of the Dunning tumor, the male accessory sex glands (ventral, dorsal, lateral prostate, coagulating gland, bulbourethral gland) and the mammary gland of both Copenhagen and Wistar rats. Mono- and polyclonal antibodies directed against intermediate filament proteins (cytokeratin, desmin, vimentin) and the contractile proteins (-actin, muscle type specific myosin, tropomyosin) were used along with phalloidin decoration of F-actin. As in the human prostate, none of the rat prostate lobes in either strain did contain basal cells expressing cytokeratin along with -actin, myosin and tropomyosin. Cells representing fully differentiated myoepithelial cells, however, were present as anticipated in the mammary gland, the bulbourethral gland and the H-tumor line of the Dunning tumor. This finding is difficult to reconcile with the contention of a prostatic origin of the H-Dunning tumor. Further studies are required to classify the epithelial parental tissue in order to define the true origin of the H-Dunning tumor and the tumor lines derived thereof.     

Modern brachytherapy for localized prostate cancers: the northwest hospital (Seattle) experience

Modern ultrasound-guided prostate brachytherapy is rapidly changing the way localized prostate cancer is managed. With routine use of prostate-specific antigen screening, prostate cancer is being diagnosed in younger men, who are understandably concerned about the morbidity of radical treatments that may significantly decrease their quality of life. Numerous studies of prostate brachytherapy have shown the excellent disease control rates achieved while maintaining low levels of urinary and erectile difficulties. This report examines a modern implant method of brachytherapy; describes patient selection for brachytherapy, alone and in combination with external beam therapy; and presents results from a series of men followed for 12 years.     

Troubles sexuels après curiethérapie par implants permanents d’I-125 dans le cancer localisé de prostate

Brachytherapy by permanent implants is an alternative to radical prostatectomy or external beam radiotherapy for good prognosis localized prostate cancer. The advantages of this treatment are effective and precise irradiation, limited to the prostate gland with moderate and transient morbidity. Erectile dysfunction, frequent erection after surgery and external beam radiotherapy, is observed in 6% to 61% of cases in the literature after brachytherapy. This wide range is related to differences in terms of follow-up, definition of sexual disorders, and the measuring instruments used. These erectile disorders occur between 9 and 17 months after treatment and appear to be related to vascular radiation lesions of the erectile bodies close to the prostatic apex (urethral bulb and base of the corpora cavernosa). However, the majority of erectile disorders respond favourably to oral treatments such as yohimbine or sildenafil. Among the various curative treatment options for localized prostate cancer, permanent implant brachytherapy is the treatment ensuring the best preservation of erectile function.     

Permanent prostate brachytherapy: the significance of postimplant dosimetry

Recent developments in imaging science and treatment-planning software allow for accurate postimplant dosimetric assessment in all patients after prostate brachytherapy. This article reviews the available data correlating cancer control and morbidity with dosimetric quantifiers obtained from postimplant dosimetric assessment after prostate brachytherapy. Continued collection of dosimetric data in patients treated with prostate brachytherapy will allow for further refinements in the technique, leading to continued high rates of cure with increasingly lower rates of morbidity.     

Polyglutamylated Tubulin Binding Protein C1orf96/CSAP Is Involved in Microtubule Stabilization in Mitotic Spindles

The centrosome-associated C1orf96/Centriole, Cilia and Spindle-Associated Protein (CSAP) targets polyglutamylated tubulin in mitotic microtubules (MTs). Loss of CSAP causes critical defects in brain development; however, it is unclear how CSAP association with MTs affects mitosis progression. In this study, we explored the molecular mechanisms of the interaction of CSAP with mitotic spindles. Loss of CSAP caused MT instability in mitotic spindles and resulted in mislocalization of Nuclear protein that associates with the Mitotic Apparatus (NuMA), with defective MT dynamics. Thus, CSAP overload in the spindles caused extensive MT stabilization and recruitment of NuMA. Moreover, MT stabilization by CSAP led to high levels of polyglutamylation on MTs. MT depolymerization by cold or nocodazole treatment was inhibited by CSAP binding. Live-cell imaging analysis suggested that CSAP-dependent MT-stabilization led to centrosome-free MT aster formation immediately upon nuclear envelope breakdown without γ-tubulin. We therefore propose that CSAP associates with MTs around centrosomes to stabilize MTs during mitosis, ensuring proper bipolar spindle formation and maintenance.     

Is tissue harmonic ultrasound imaging (THI) of the prostatic urethra and rectum superior to brightness (B) mode imaging? An observer study

Quality ultrasound images are an essential part of prostate brachytherapy procedure. The authors have previously reported that tissue harmonic ultrasound images (THI) are superior to brightness (B) mode for the prostate. The objective of the current study was to compare both imaging modes for visualization of the prostatic urethra and rectum.B and THI mode transrectal ultrasound images were acquired for ten patients. The prostatic urethra and rectal wall were contoured by a radiation oncologist (RO) and five observers on randomly presented images. The contours on one patient were repeated four additional times by four observers. All the images were qualitatively scored using a five-level Likert scale.The values of the Pearson product-moment correlation coefficients showed that the observers were in close agreement with the RO. Two sample paired student t-test showed that the rectum volumes with THI were significantly smaller than B-mode, but no significant difference for urethra. Two-factor analysis of variances showed significant observer variability in defining the rectum and urethra in both imaging modes. Observer consistency of the rectum volumes, estimated by standard deviations as percentages of means was significantly improved for THI. The Likert scale based qualitative assessment supported quantitative observations.The significant improvement in image quality of the prostate (reported previously) and rectum with THI may offer better-quality treatment plans for prostate brachytherapy and potential improvement in local control.     

A novel cadmium induced protein in wheat: characterization and localization in root tissue

A 51-kDa soluble protein was over-expressed in wheat (Triticum aestivum) seedlings by the treatment of seeds before germination with 50 μM CdCl₂ for 48 h and subsequently washed off Cd²⁺. This protein designated as Cd stress associated protein (CSAP), was purified. Polyclonal antibody was raised against CSAP for localizing the protein in root tissue of treated and control seedlings. It was observed that CSAP was located below the plasma membrane and outer periphery of the tonoplast. This unique type of organized localization of CSAP is suggestive of defensive role against metal phytotoxicity. N-terminal analysis of CSAP and expressed sequence tags (EST) database search of wheat sequences suggests that this protein has not been reported earlier in higher plants.     

Age-dependent expression of 5alpha-reductase and androgen receptors mRNA by the canine prostate

The growth of the prostate gland is androgen-dependent. Testosterone is converted to the most potent dihydrotestosterone (DHT) by 5alpha-reductase within the prostate. Androgen interacts with androgen receptors (AR) to regulate normal growth of the prostate and has also been implicated in both the progression of benign prostate hyperplasia and prostate cancer. This study was conducted to compare the mRNA expression of AR and 5alpha-reductase by the prostate gland from three age categories: immature, young-mature and old dogs. Quantitative gene expression was assessed by the real-time PCR and the results were expressed as a relative mRNA expression of the target gene. This study revealed that there was no significant difference in the mRNA expression of the AR gene by the prostate gland of immature, young and old dogs. In contrast, there is a highly significant (P<0.001) down-regulation in 5alpha-reductase gene by the prostate of young and old dogs as compared with immature dogs. However, there is no significant difference in mRNA expression of the 5alpha-reductase gene by the prostate gland from young and old dogs. This differential expression of AR and 5alpha-reductase genes, which are involved in the regulation of androgen effect on prostate gland, might reflect an age-dependent growth requirement of the gland for androgens.     

A genetic screen in Drosophila for regulators of human prostate cancer progression

To uncover the mechanism by which human prostate cancer progresses, we performed a genetic screen for regulators of human prostate cancer progression using the Drosophila accessory gland, a functional homolog of the mammalian prostate. Cell growth and migration of secondary cells in the adult male accessory gland were found to be regulated by paired, N-cadherin, and E-cadherin, which are Drosophila homologues of regulators of human prostate cancer progression. Using this screening system, we also identified three genes that promoted growth and migration of secondary cells in the accessory gland. The human homologues of these candidate genes – MRGBP, CNPY2, and MEP1A – were found to be expressed in human prostate cancer model cells and to promote replication and invasiveness in these cells. These findings suggest that the development of the Drosophila accessory gland and human prostate cancer cell growth and invasion are partly regulated through a common mechanism. The screening system using the Drosophila accessory gland can be a useful tool for uncovering the mechanisms of human prostate cancer progression.