Stable knockdown of CREB, HIF-1 and HIF-2 by replication-competent retroviruses abrogates the responses to hypoxia in hepatocellular carcinoma.

The fast proliferation of tumor cells develops faster than the vasculature, resulting, in most malignant tumors, in generation of hypoxic regions. Hypoxia renders solid tumors resistant to radiation and chemotherapeutics while providing opportunities for tumor-selective therapies targeting tumor hypoxia. Here we exploit two properties of tumors: propagation of tumor cells and ongoing generation of hypoxic regions to construct a system that preferentially leads to the death of tumor cells and thus hinders tumor growth. We constructed murine leukemia virus replication-competent (RCR) viruses that infect only propagating cells. These viruses express small hairpin RNAs (shRNAs) targeting cyclic AMP-response-element binding protein (CREB), hypoxia-inducible factors 1 (HIF)-1 or HIF-2 individually or all three together (X3). These viruses efficiently infected in vitro human hepatocellular carcinoma (HepG2 and FLC4) cells and established persistence of the virus and knocked down the expression of the regulators of the hypoxia-responding genes. Knockdown of either HIF-1 or CREB or both in hypoxia reduced the expression of hypoxia-response elements- and CRE-mediated gene expression, diminished cell proliferation and increased caspase-3 activity. We did not detect any significant effect of the efficiently knocked down HIF-2 on any of the functions tested in vitro. Moreover, severe combined immunodeficiency mice implanted subcutaneously with HepG2 stably infected with recombinant RCRs showed reduction of tumor growth and vascular endothelial growth factor expression, and no hypoxia-guided neovascularization. Combined treatment (RCRs+doxorubicin) improved efficacy in the context of in vitro hypoxia and in vivo (with either vACE-CREB or vACE-X3). This synergistic effect may lead to an improved efficacy and safety profile of the treatment that may result in fewer side effects.

Bipolar versus monopolar transurethral resection of the prostate for benign prostatic hyperplasia: safe in patients with high surgical risk.

Here, we compared the effects of bipolar and monopolar transurethral resection of the prostate (B-TURP, M-TURP) for treating elderly patients (≥75 years) with benign prostatic hyperplasia(BPH) who had internal comorbidities. Eligible BPH patients were aged ≥75 years and had at least one internal comorbidity. In this open-label, prospective trial, patients were assigned to B-TURP (n = 75) and M-TURP (n = 88) groups. Data on prostate volume (PV), urination, and time during perioperative period were compared; data associated with urination and complications at one year postoperatively were also compared. Finally, follow-up data were available for 68 and 81 patients in the B-TURP and M-TURP group, respectively. No deaths were recorded. Intraoperative bleeding was lower and irrigation time, indwelling catheter time, and hospital stay were shorter in the B-TURP group than in the M-TURP group (p < 0.001). No difference was observed with respect to operation time (p = 0.058). At one year after the operation, differences with respect to urination and complications were not significant. In conclusion, Short-term efficacy of B-TURP or M-TURP was satisfactory for elderly patients with BPH who had internal comorbidities. Besides, B-TURP is a more sensible choice because it has a lower prevalence of adverse effects.

A diffusion tensor magnetic resonance imaging study of corpus callosum from adult patients with migraine complicated with depressive/anxious disorder.

OBJECTIVE: The aim of this study was to investigate the possible microstructural abnormalities of the corpus callosum (CC) in adult patients with migraine without aura complicated with depressive/anxious disorder. BACKGROUND: Emotional disorders, especially depression and anxiety, are with relatively higher incidence in migraine population. However, the mechanism of migraine complicated with depressive/anxious disorder remains unclear. METHODS: Diffusion tensor magnetic resonance imaging was carried out in 12 adult patients with simple migraine (without aura and without depressive/anxious disorder) (S-M group), 12 adult patients with complicated migraine (without aura but complicated with depressive/anxious disorder) (Co-M group), and 12 age- and sex-matched healthy subjects (Control group). Fractional anisotropy (FA) and apparent diffusion coefficient were measured at genu, body, and splenium of the CC, respectively. RESULTS: There were significant differences in FA values at all locations of the CC among the 3 groups. The FA values from both the SM and Co-M groups were significantly lower than the control (P < .05 and P < .01, respectively). The FA values from Co-M group were significantly lower than the SM group (P < .01). The apparent diffusion coefficient values of the above regions had no significant differences among these groups (P > .05). There were negative correlations between FA value of genu of the CC and disease course as well as FA value of genu and body of the CC and headache frequency (P < .05). Negative correlations were also found between FA values at all locations of the CC and Hamilton anxiety and Hamilton depression scores (both P < .05). CONCLUSIONS: There might be an integrity change of neurofibrotic microstructures existing as a possible neuroanatomical basis in the CC of migraine patients complicated with depressive/anxious disorder.

Ruthenium-106 plaque brachytherapy for thick posterior uveal melanomas.

AIMS: Ruthenium-106 brachytherapy is an effective method for treating small to medium uveal melanomas. The purpose of this study was to examine its effectiveness and safety in the management of thick posterior uveal melanoma (apical height >/=8.0 mm) and to compare it with enucleation. METHODS: 126 consecutive patients with thick uveal melanoma were included. 63 patients treated with Ruthenium-106 brachytherapy were compared with 63 patients treated with enucleation. The main outcome measures were visual acuity, eye retention, local recurrence, metastases, all-cause mortality and melanoma-related mortality. RESULTS: Patients treated with brachytherapy were significantly younger and had significantly smaller tumours, compared with patients treated with enucleation. Tumour thickness in the brachytherapy group was 9.3 (SD 0.9) mm compared with 12.2 (1.9) mm in the enucleation group. The 5- and 10-year melanoma-related mortality was 20.5% and 46.2% for brachytherapy patients and 28.1% and 44.0% for enucleation patients (p = 0.6 and p = 0.9). When comparing 15 brachytherapy with 15 matched enucleation patients, the 5-year melanoma-related mortalities were similar, 28.6% and 33.3% respectively (p = 0.7). Complications associated with brachytherapy included tumour regrowth (n = 15), scleral melt (n = 3), neovascular glaucoma (n = 5) and vitreous haemorrhage (n = 3). In the brachytherapy group, no significant difference in survival was noted between patients who did and did not develop local recurrence (p = 0.9). Of the eyes that were initially treated with brachytherapy, 71.4% were saved from enucleation. Of these, 70.8% had a final visual acuity of 20/200 or better. CONCLUSIONS: Ruthenium-106 brachytherapy is an alternative to enucleation in some thick posterior uveal melanomas.

Long-term survival of uveal melanoma patients after surgery for liver metastases.

AIMS: To evaluate the posthepatectomy survival of uveal melanoma patients with liver metastases. METHODS: Data were collected from the files in the Departments of Ophthalmology, General Surgery and Oncology, for uveal melanoma patients who were seen in the Ocular Oncology Clinic at the Hadassah Medical Center from 1988 to 2007. The main outcome was posthepatectomy survival. Statistical analysis was performed using JMP statistical software. RESULTS: Of the 558 patients, 74 (13%) developed metastases after a median of 35.0 months from the initial diagnosis. Thirty-five patients underwent hepatectomy. These patients had similar clinical characteristics as those who did not undergo hepatectomy. The median survival time from the detection of metastasis was 3.7-fold higher in the operated patients in comparison with the non-operated patients. Posthepatectomy survival of patients who were found in surgery to have 1-5 metastatic nodules was 3.1 times longer than those with six or more lesions. The hepatectomies of 13 patients resulted in complete resection of the hepatic metastases with clean histological margins (R0). These patients survived 1.9 times longer than those with residual disease (R1/R2). CONCLUSION: It is possible to extend significantly the life expectancy of uveal melanoma patients who develop isolated hepatic metastases by complete resection of the lesions.

Retinal nerve fibre layer imaging compared with histological measurements in a human eye.

PURPOSE: A feasibility study comparing retinal nerve fibre layer (RNFL) thickness values obtained with imaging devices against RNFL thickness measurements obtained histologically in a human eye. DESIGN: A single patient scheduled for orbital exenteration, who still possessed a healthy functioning eye. METHODS: Before surgery, the eye was imaged using optical coherence tomography (OCT) and scanning laser polarimetry (SLP). After orbital exenteration, the globe was sectioned, and 100 equidistant RNFL thickness measurements were obtained for each of four concentric rings centred on the optic disc, with diameters of 3.0, 3.5, 4.0, and 4.5 mm. RESULTS: RNFL thickness was found to be inversely related to the distance from the centre of the optic disc along each radial meridian. Peripapillary RNFL thickness was found comparable for histology, OCT, and SLP. RNFL thickness measured histologically confirmed a 'double hump' pattern, peaking at the superior and inferior poles. Histologically derived RNFL thickness (microm), at 3.0 and 3.5 mm diameter ring ranged between 30-135 and 25-115 respectively. In comparison, the 3.0 mm diameter GDx data ranged between 25 and 100, and the 3.4 mm diameter OCT data between 40 and 175. CONCLUSIONS: Imaging data appear qualitatively similar when compared to the histologically derived data. Quantitative differences may be partly due to scaling differences and histological artefacts. The histological analysis approach demonstrated in this study can potentially serve to validate imaging-derived data, as well as help improve our understanding of RNFL loss in glaucoma.

Lesions of the caruncle: a description of 42 cases and a review of the literature.

Lesions of the caruncle are uncommon. On account of the histological composition of the caruncle, which includes, in addition to conjunctiva, hair follicles, sebaceous glands, sweat glands, and accessory lacrimal tissue, the caruncle may develop lesions that may be similar to those found in the skin, conjunctiva, or lacrimal gland. Clinical preoperative diagnosis is very difficult and reached only in about half of the cases. The vast majority of lesions of the caruncle are benign, mainly nevi. Reported malignant lesions are very rare but can be potentially fatal. Although malignancy is clinically overestimated, any suspected malignant lesion should be excised and examined histopathologically by an experienced pathologist. This study presents the clinical and histological data of 42 consecutive caruncular lesions processed at our laboratory and reviews previously reported cases of caruncular lesions.

Intravitreal methotrexate for treating vitreoretinal lymphoma: 10 years of experience.

AIM: To describe our experience in treating vitreoretinal involvement of primary central nervous system lymphoma, by intravitreal injections of methotrexate (MTX). METHODS: Patients with suspected intraocular lymphoma underwent a diagnostic vitrectomy. Samples were sent for cytology, genetic evaluation and for interleukin level measurements. Treatment protocol included injection of 400 microg/0.1 ml MTX intravitreally twice weekly for 4 weeks, once weekly for 8 weeks, and then once monthly for 9 months, for a total of 25 injections. Data were collected from the patients' records and included, inter alia, response to intravitreal MTX measured by time to disappearance of vitreal cells and retinal infiltrates, changes in visual acuity, and clinical recurrence rate. RESULTS: In the past 10 years we have treated 44 eyes of 26 patients; seven patients had monocular involvement, and 19 binocular. Six patients were initially diagnosed as having a non-responsive uveitis, and 16 with either CNS or systemic lymphoma with later involvement of the eye. Four patients had systemic lymphoma; one of them was found to have CNS lymphoma after the ocular involvement. Three patients had T cell lymphoma, and the rest had B cell lymphoma. Clinical remission was reached after 6.4 (3.4) (2-16) injections of MTX (mean (SD) (range)), with 95% of the eyes needing 13 injections or less to be cleared of malignant cells. None of the patients had an intraocular recurrence. Among the side effects, the most common was corneal epitheliopathy, which usually appeared after the third injection and began to subside when the intervals between injections increased. CONCLUSIONS: Vitreoretinal involvement of lymphoma can be controlled effectively and without serious adverse reactions by intravitreal MTX injections. The treatment protocol described herein has resulted in no intraocular recurrence so far and has had bearable side effects. The accumulating clinical results bring us to propose the consideration of this protocol as a good first-line treatment option for intraocular lymphoma.

Maculopathy in patients with primary CNS lymphoma treated with chemotherapy in conjunction with blood-brain barrier disruption.

AIM: To determine the incidence and characteristics of maculopathy associated with blood-brain barrier disruption (BBBD) which is used in treating primary central nervous system lymphoma (PCNSL). METHODS: Files of all 56 patients with PCNSL, with or without intraocular lymphoma (IOL), treated at Hadassah University Hospital during the years 1997-2007 were reviewed. Data on 46 patients for whom we had documentation of ocular examination were studied. Those who were alive at the time of the data collection were invited for further evaluation of the presence or absence of maculopathy. The patients were divided into four groups according to treatment protocol. Group 1: systemic intravenous chemotherapy; Group 2: systemic intravenous chemotherapy and intravitreal methotrexate (MTX); Group 3: systemic intra-arterial (IA) chemotherapy and BBBD; Group 4: systemic IA chemotherapy, BBBD, and intravitreal MTX. RESULTS: Of the 23 patients of Groups 1 and 2 who were not treated by BBBD, none developed maculopathy. Of those 23 patients who were treated by BBBD, 12 of 17 (70.5%) in Group 3 and three of six (50%) in Group 4, for a total of 15 of 23 (65.2%) developed maculopathy. The maculopathy did not significantly affect visual acuity in any of them. CONCLUSIONS: BBBD may cause maculopathy in almost two-thirds of patients treated for PCNSL, without affecting significantly the visual acuity. Intravitreal injection of MTX, according to the protocol which we apply, is not associated with maculopathy.

Demonstrating circulation in vasculogenic mimicry patterns of uveal melanoma by confocal indocyanine green angiography.

PURPOSE: Vasculogenic mimicry patterns, formed by highly invasive melanoma cells, connect to endothelial cell-lined blood vessels and contain fluid in vitroand in vivo. This study was designed to determine if fluid leaks into vasculogenic mimicry patterns without circulation, or if fluid circulates in and clears from these patterns. METHODS: Indocyanine green (ICG) laser scanning confocal angiography (Heidelberg Retinal Angiograph (HRA); Heidelberg Engineering, Heidelberg, Germany) was performed on nine patients with posterior choroidal melanoma in an institutional setting. Blood was drawn before the ICG injection and from the contralateral arm of the ICG injection site and 1 min after the injection. Outcome measures include time to first filling of retinal vessels and vasculogenic mimicry patterns and the time at which no fluorescence could be detected by the HRA instrument. After fluorescence was no longer detected in vessels or patterns, the tubes containing the patient's blood was imaged by the Heidelberg HRA. RESULTS: Looping vasculogenic mimicry patterns were detected focally in five patients within 30 s after injection and were detectable up to 12 min post-injection. Blood drawn before ICG injection did not autofluoresce but ICG-containing blood pooled in the tube continued to fluoresce at 1-month post-injection. CONCLUSIONS: Vasculogenic mimicry patterns are not part of the endothelial cell-lined vascular system and fluid enters these patterns through leakage. The rapid infusion of ICG into these patterns after injection and the disappearance of fluorescence detectable by the Heidelberg HRA suggest that fluid circulates in these patterns and does not accumulate as a stagnant pool.