Inverse co-expression of EZH2 and acetylated H3K27 in prostatic tissue.

CONTEXT: Enhancer of Zeste 2 (EZH2), a methyltransferase and an upregulated gene is an adverse prognosticator in prostate cancer. It catalyzes histone H3 lysine 27 trimethylation (H3K27me3) leading to repressive chromatin status (heterochromatin). Following demethylation and acetylation of H3 protein (H3K27ac) the result is transcriptionally activated status (euchromatin), a key metastasis facilitator being targeted by ongoing clinical trials, as with palbociclib. Here, we performed the first immunohistochemical study of H3K27ac expression in prostatic tissue and cancer metastasis, and determined a possible correlation with EZH2 expression. METHODS: Tissue microarrays were made and immunohistochemistry was performed for EZH2 and H3K27ac. Slides were scanned and image data utilized a software-assisted, unbiased quantification method. The software captured diaminobenzidine positive regions, and tissue areas. RESULTS: Benign prostate tissue expressed almost no EZH2 but showed strong H3K27-Ac positivity. Tumor was EZH2 positive (p < 0.05 vs. benign) with strongest staining in lymph node metastasis. H3K27-Ac was decreased in tumors, yet paradoxically had stagewise and gradewise progressive increases (both p < 0.05), with the strongest staining in lymph nodes. The overall relationship of EZH2 and H3K27ac was weakly correlated (r = 0.28, p < 0.05). CONCLUSIONS: EZH2 and H3K27ac had an inverse correlation in benign versus (especially) low-grade and low-stage prostate cancers; however, in high-stage and high-grade cancers and metastases, H3K27ac increased significantly. Findings support EZH2 and H3K27ac as targets for cancer prevention in localized or low-grade prostate cancer, but we now note that their inverse relationship becomes uncoupled in advanced prostate cancer.

Small Cell Variant of Metastatic Melanoma: A Mimicker of Lymphoblastic Leukemia/Lymphoma.

It is well-known to pathologists that melanoma is "the great mimicker" and can look like anything. Despite this widespread awareness, the diagnosis remains a continuous challenge, especially when a metastatic melanoma with rare morphology is examined. We report a case of a 64-year-old man with a lung mass and right-sided pleural effusion who underwent video-assisted thoracoscopic surgery for pleural decortication. The history of melanoma was not reported to us. Microscopic examination revealed sheets of small round blue cells infiltrating into the adipose tissue in a lace-like pattern mimicking lymphoblastic lymphoma. Immunohistochemical stains for melanocytic markers, including S-100 protein, Mart-1, and HMB-45, highlighted the neoplastic cells. The tumor was also positive for CD56 and CD117, but negative for pancytokeratin, CD45, cytokeratin 8, TTF-1, WT1, CD34, chromogranin, synaptophysin, and neuron-specific enolase. The findings were most consistent with metastatic small cell melanoma, an uncommon variant of melanoma that closely resembles lymphoblastic lymphoma and other malignant small round blue cell tumors. To our knowledge, we are the first to describe a case of metastatic small cell melanoma to the pleura in an adult. Clinical and histological details are provided with a review of the literature.

Patients' attitude towards travelling for breast services versus waiting longer for local services

Geographical access is a cornerstone of health care provision. However, prolonged waiting for breast clinic appointments in local services results in delayed diagnosis and excessive anxiety for patients. In this study, we present a patient satisfaction survey results of an initiative to offer out-patient clinic appointments for non-urgent patients referred to the breast unit in Letterkenny General Hospital (LGH), Ireland, and exceeded the recommended waiting period of 12 weeks. These patients were offered appointment in University Hospital Galway (UCHG), which is an average travel time of about 3.5-4.5 hours away from LGH. 163 patients out of 336 (48.5%) patients actively waiting more than 12 weeks for appointments in LGH accepted alternative appointments in UCHG. Despite the long travel distance for these patients, 100% of them reported high satisfaction and 97.3% said they would accept further UCHG appointments if a similar situation of prolonged waiting in LGH arises. None of these patients were diagnosed with cancer, and only one had a benign lumpectomy. This study showed that if offered alternative appointments, just under half of the patients would accept. The initiative provides a feasible solution for long waiters, and the survey shows that patients' satisfaction remains high despite long travel.

Sequential combined treatment with allopurinol and benznidazole in the chronic phase of Trypanosoma cruzi infection: a pilot study.

OBJECTIVES: Even though the use of combined drugs has been proved to be effective in other chronic infections, assessment of combined treatment of antiparasitic drugs in human Chagas' disease has not been performed. Herein, a pilot study was conducted to evaluate the tolerance and side effects of a sequential combined treatment of two antiparasitic drugs, allopurinol and benznidazole, in the chronic phase of Trypanosoma cruzi infection. PATIENTS AND METHODS: Changes in total and T. cruzi-specific T and B cells were monitored during a median follow-up of 36 months. Allopurinol was administered for 3 months (600 mg/day) followed by 30 days of benznidazole (5 mg/kg/day) in 11 T. cruzi-infected subjects. RESULTS: The combined sequential treatment of allopurinol and benznidazole was well tolerated. The levels of T. cruzi-specific antibodies significantly decreased after sequential combined treatment, as determined by conventional serology and by a multiplex assay using recombinant proteins. The frequency of T. cruzi-specific interferon-γ-producing T cells significantly increased after allopurinol treatment and decreased to background levels following benznidazole administration in a substantial proportion of subjects evaluated. The levels of total naive (CD45RA + CCR7 + CD62L+) CD4 + and CD8 + T cells were restored after allopurinol administration and maintained after completion of the combined drug protocol, along with a decrease in T cell activation in total peripheral CD4 + and CD8 + T cells. CONCLUSIONS: This pilot study shows that the combination of allopurinol and benznidazole induces significant modifications in T and B cell responses indicative of a reduction in parasite burden, and sustains the feasibility of administration of two antiparasitic drugs in the chronic phase of Chagas' disease.

Papillary cystadenofibroma of epididymis: a case report.

We present the first reported case of papillary cystadenofibroma of the epididymis. The tumor occurred in a 46-year-old man. The mass was 3.7 cm and included a hemorrhagic fluid-filled cyst. Microscopically, stromal-filled papillae were lined by low cuboidal to columnar epithelium. Epithelial cells were reactive for cytokeratin 7, cytokeratins AE1/3, and focally in the apical cytoplasm for CD10. Focal CD10 reactivity was also noted in the stroma. The lesion was negative for alpha-fetoprotein. These findings ruled out other lesions, including metastatic renal cell carcinoma.

Leg pain in the running athlete.

Leg pain is a common complaint among recreational and professional athletes who compete in running sports. Evaluation of the individual with intermittent or constant leg pain should be well organized and inclusive. Duration of the pain, its relation to injury, intensity of the pain, and its pattern are important factors. Additionally, changes in the training regimen, its level, intensity, or duration, or in the nature of the routine are critical components of the assessment. Physical examination can help differentiate bony from soft-tissue etiologies. Studies are dictated by the differential diagnosis but include radiographs in almost all patients and selected use of other modalities. These include bone scans and magnetic resonance imaging for medial tibial stress syndrome and stress fractures and intracompartmental pressure measurements for chronic compartment syndrome. Treatment often requires either rest or a change in training regimen. Surgery for conditions such as chronic compartment syndrome frequently allows a return to preinjury activities.

Androgen ablation therapy for prostate carcinoma suppresses the immunoreactive telomerase subunit hTERT.

BACKGROUND: Telomerase is a ribonucleoprotein complex that protects the ends of chromosomes from degradation. Its catalytic subunit, hTERT, controls its activity. Prior data in prostate carcinoma cases indicated that immunohistochemical hTERT reactivity increases with tumor grade and may be absent in lower grade cases. The effect of complete androgen ablation (CAA) on tumor hTERT expression was uncertain. METHODS: hTERT immunostaining was performed on the cancerous pretreatment biopsy tissue of 30 men who consecutively underwent CAA with bicalutamide and goserelin acetate for 30 days prior to undergoing radical prostatectomy, and on their tumor tissue from radical prostatectomy. As controls, biopsy and prostatectomy samples from 30 untreated men were studied. Nuclear staining was evaluated by two observers, and the change in staining between biopsy and prostatectomy samples was evaluated using the Student t test in both groups. RESULTS: The percent of reactive tumor nuclei in treated men declined from 36.7% to 13.2% (P = 0.0001), and declined from 19.8% to 16.1% in untreated men (P = 0.4). The greater mean hTERT reactivity in the treated men's biopsy specimens was attributed to an increased proportion of higher (Gleason score > or = 7) grade tumors. The decline in hTERT immunostaining remained significant after normalizing it to that of the untreated group (P = 0.002). The original Gleason scores, corresponding declines in the percentage of reactive tumor nuclei, and significance were: Gleason score < or = 6: 11% (P = 0.03); Gleason score of 7: 23% (P < 0.006); and Gleason score > or = 8: 46% (P < 0.005) (from a mean 63% to 17%). CONCLUSIONS: CAA for prostate carcinoma can be considered an antitelomerase therapy. The steepest reduction in telomerase activity was noted in the highest grade tumors.

Prostate cancer overexpresses CD44 variants 7-9 at the messenger RNA and protein level.

BACKGROUND: In prostate cancer, prior data show down-regulated immunohistochemical expression of cell adhesion protein CD44 standard (CD44s) and most variants (CD44v). MATERIALS AND METHODS: Expression of CD44 mRNA was studied by RT-PCR and cDNA sequencing in 19 prostate cancers and 10 benign controls. Immunohistochemical staining was performed with anti-CD44v7/8 in 80 prostatectomy specimens, and 12 were used for in situ hybridization for CD44v7 (exon 12). Western blotting with monoclonal antibody to CD44 standard, v6, v7/8, or v9 was performed using cancerous and benign prostate. RESULTS: Sequencing of RT-PCR products showed that benign tissue and cancer express CD44 standard and v10 mRNA at 482 base pairs (bp). In contrast, cancer tissues also overexpressed 800-1000 bp bands corresponding to v7-9 isoforms. In situ hybridization revealed increased CD44v7 signal in cancer compared to benign acini. Immunostaining for CD44v7/8 was increased, proportional to Gleason grade. By Western blot, cancer and benign tissue disclosed major bands of reactivity at 75-100 kD consistent with intact standard and variant isoforms. Tumors, however, had 6-45 kD bands for CD44 standard, v7/8 and v9, consistent with cleavage products. CONCLUSION: CD44 v7-9 isoform messenger RNA is increased in prostate cancer, and translation yields low molecular weight polypeptides of probable cleavage origin.

Overexpression of NSAID-activated gene product in prostate cancer.

NSAID-activated gene (NAG-1) protein was previously identified by microarray analysis as overexpressed in prostate cancer. We performed immunohistochemistry and Western blotting with rabbit polyclonal antibody to NAG-1. Fifty malignant tissues obtained by prostatectomy and 17 from benign cases were compiled. Cancer tissues included Gleason scores 3-6, 3+4=7, 4+3=7, and 8-10. Cancer and high-grade prostatic intraepithelial neoplasia (PIN) consistently showed moderate to intense cytoplasmic reactivity in 95-100% of epithelium. Staining intensity inversely correlated with preoperative serum prostate-specific antigen (PSA) (p=0.005) and with grade, averaging (on a 0 to 3+ scale) 2.3 +/- 0.6 in the lowest grade group, and 2.0 +/- 0.7, 1.8 +/- 0.5, and 1.5 +/- 0.6 as grade increased (p<0.008). Benign epithelium was nonreactive in 17/17 specimens without concurrent cancer (11 transurethral resection, 2 enucleation, 4 biopsy, p=0.002). Decreased NAG-1 expression in higher grade cancer is consistent with its known antitumorigenic, proapoptotic activities.

Lobular carcinoma of the breast metastatic to the epidural space: a potential mimic of giant cell bone tumor.

We report the case of a 64-year-old woman with a spinal epidural mass. Tissue from a decompression laminectomy disclosed a tumor with numerous osteoclast-like giant cells separated by small, moderately atypical tumor cells. The osteoclast-like giant cells were immunoreactive for vimentin, but negative for epithelial membrane antigen and broad-pectrum cytokeratin. Subsequent breast biopsy revealed a lobular carcinoma of classic type without osteoclast-like giant cells. This is the first reported case in which metastatic breast carcinoma was accompanied by these giant cells but the giant cells were not present at the primary tumor site.

Telomerase reverse transcriptase subunit immunoreactivity: a marker for high-grade prostate carcinoma.

BACKGROUND: Telomerase, a ribonucleoprotein complex that maintains telomeric DNA, has been detected in 67-93% of prostate carcinomas by telomeric repeat-amplification protocol assay (involving polymerase chain reaction). One study used in situ hybridization in nine patients; however, to date, no immunohistochemical results have been published. METHODS: From two hospitals, the authors compiled data on 62 patients who underwent prostatectomy from January 1996 to May 2001. Representative tissue sections were immunostained with a polyclonal antibody to telomerase reverse transcriptase (TERT), the catalytic subunit of telomerase. Staining was evaluated by two observers and was correlated with grade, stage, and biochemical failure. There were 28 sections from low-grade to intermediate-grade tumors (Gleason score, 3-6), 14 sections with a Gleason score of 3 + 4 = 7, 9 sections with a Gleason score of 4 + 3 = 7, and 11 sections from high-grade tumors (Gleason score, 8-10). RESULTS: From low-grade to high-grade tumors, the four groups described above disclosed nuclear reactivity in 64%, 100%, 100%, and 100% of sections, respectively. Mean percentages of 5%, 15%, 40%, and 51% of nuclei were reactive in the respective groups (P < 0.0001) with intratumoral heterogeneity. The percent of reactive tumor nuclei was not correlated with pathologic stage (P = 0.32) or margin status (P = 0.35). The basal cell layer in sections of high-grade prostatic intraepithelial neoplasia (HGPIN) and benign/atrophic acini was reactive; secretory cells were reactive in 13 of 34 HGPIN foci (38%) in 1-20% of nuclei and were never reactive in benign acini. Lymphocytes and skeletal muscle were reactive. Weak, nonspecific, cytoplasmic staining was noted in benign and tumor acini. CONCLUSIONS: Like cytokeratin 34betaE12, nuclear anti-TERT reactivity is a basal cell marker in nonneoplastic prostatic acini. Anti-TERT reactivity is acquired by secretory cells in tumorigenesis, but consistent reactivity is restricted to high-grade carcinoma (Gleason primary pattern >or=4). This histologic evidence suggests that higher grade tumors have maximally activated telomerase and may be most responsive to antitelomerase therapy.

Herpes simplex vegetans: atypical genital herpes infection in a patient with common variable immunodeficiency.

Large papillomatous lesions clinically resembling verrucous carcinoma may be caused by viruses other than human papillomavirus. We report a case of recurrent vegetations covering the entire vulva in a pregnant patient with common variable immunodeficiency. Herpes simplex virus was recovered from these lesions. The patient did not respond to intravenous acyclovir, but her lesions dramatically healed with two courses of intravenous foscarnet. Repeated biopsies may prove necessary in cases such as this to ensure proper diagnoses.

Improved dose localization with dual energy photon irradiation in treatment of lateralized intracranial malignancies.

Dual energy photon irradiation (6 MV and 20 MV) was compared to conventional treatment planning with 6 MV photons in a lateralized intracranial malignancy. Dose volume analysis was performed of both the tumor plus a 2 cm margin (target volume, TV) and normal tissues (NT). Parallel opposed treatment using weightings of 1:1, 1.5:1, and 2:1 were compared for 6 MV photons alone or in combination with 20 MV photons. Uniform treatment of the TV was accomplished within the 60 Gy isodose. Significant differences were observed, however, in NT volumes receiving greater than or equal to 60 Gy and 45-59 Gy. Dual photon energy reduced treatment of NT volumes to greater than or equal to 60 Gy by 13% (177 cm3 vs 204 cm3 in 2:1 weighting) to 70% (147 cm3 vs 498 cm3 in 1:1 weighting) for comparable plans. Dose optimization was also performed for both 6 MV alone or in combination with 20 MV photons. Usual approaches to achieve dose lateralization with conventional isocentric techniques were applied including parallel opposed 6 MV photons ipsilaterally weighted 3.4:1 (POP), and a 110 degrees arc rotational field used to limit treatment to the eye (ARC). Dual energy photon optimized plans included a three beam parallel opposed plan (TOP) and a mixed photon ipsilateral (IPSI) approach. The technique using parallel opposed 20 MV photons and ipsilateral 6 MV photons (TOP) used beam weightings of 1.1 (contralateral 20 MVX): 1.6 (ipsilateral 6 MVX): 1 (ipsilateral 20 MVX) to achieve dose optimization. The ipsilateral approach with 6 MVX and 20 MVX (IPSI) used beam weightings of 1:1.4, respectively. All optimized plans demonstrated a 41% (120 cm3; POP) to 53% (95 cm3; TOP) improvement over parallel opposed 6 MV photons weighted 2:1 (204 cm3) in NT volume receiving greater than or equal to 60 Gy. Comparison of optimized treatment showed the IPSI plan to be superior, treating 12% of NT volume to greater than or equal to 60 Gy and 38% to 45-59 Gy; the 6 MV POP plan resulted in NT volumes of 15% and 51%, respectively, for those dose levels. Dual photon energy irradiation of lateralized intracranial malignancies allows reduction of dose to normal tissue volumes while achieving excellent coverage of the target volume. Treatment planning should be performed in all lateralized intracranial lesions to achieve dose optimization exploiting depth dose characteristics.

The identification of the weak oestrogen equol [7-hydroxy-3-(4'-hydroxyphenyl)chroman] in human urine.

The identification (by gas chromatography-mass spectrometry and n.m.r.) for the first time of the weak oestrogen equol [7-hydroxy-3-(4'-hydroxyphenyl)chroman] in human urine is described. Preliminary results of its quantitative excretion in urine are reported and the potential significance of the occurrence of this compound is discussed.

The definitive identification of the lignans trans-2,3-bis(3-hydroxybenzyl)-gamma-butyrolactone and 2,3-bis(3-hydroxybenzyl)butane-1,4-diol in human and animal urine.

The definitive identification of the first lignans to be found in humans and animals is described. Gas chromatography--mass spectrometry, n.m.r. spectroscopy, i.r. spectroscopy and chemical techniques were employed to establish the structures of two lignans as trans-2,3-bis(3-hydroxybenzyl)-gamma-butyrolactone and 2,3-bis(3-hydroxybenzyl)butane-1,4-diol. Both compounds are essetially racemic. Evidence was also found for several methoxy analogues of these lignans in the vervet monkey.

An improved synthesis of 6beta-hydroxy-5beta-pregnane-3,20-dione.

6beta-Hydroxy-5beta-pregnane-3,20-dione, formerly prepared by hydroboration method, has been obtained in greatly improved yield by a simpler irradiation-hydrogenation procedure.

Effect of pentagastrin infusion on gastroesophageal manometry and reflux status before and after esophagogastrectomy.

The lower esophageal high pressure zone of eight Rhesus monkeys was characterized by manometry and reflux testing. It was found to respond characteristically to pentagastrin infusions. After excision of the lower esophageal segment and tis replacement by a stomach tube, an apparently normal, functional high pressure zone was demonstrable within the substitute.