Zirconium catalyzed synthesis of 2-arylidene Indan-1,3-diones and evaluation of their inhibitory activity against NS2B-NS3 WNV protease.

A simple and efficient Knoevenagel procedure for the synthesis of 2-arylidene indan-1,3-diones is herein reported. These compounds were prepared via ZrOCl2·8H2O catalyzed reactions of indan-1,3-dione with several aromatic aldehydes and using water as the solvent. The 2-arylidene indan-1,3-diones were obtained with 53%-95% yield within 10-45 min. The synthesized compounds were evaluated as inhibitors of the NS2B-NS3 protease of West Nile Virus (WNV). It was found that hydroxylated derivatives impaired enzyme activity with varying degrees of effectiveness. The most active hydroxylated derivatives, namely 2-(4-hydroxybenzylidene)-1H-indene-1,3(2H)-dione (14) and 2-(3,4-dihydroxybenzylidene)-1H-indene-1,3(2H)-dione (17), were characterized as noncompetitive enzymes inhibitors, with IC50 values of 11 µmol L-1 and 3 µmol L-1, respectively. Docking and electrostatic potential surfaces investigations provided insight on the possible binding mode of the most active compounds within an allosteric site.

Secretome derived from different cell lines in bovine embryo production in vitro.

The present study investigated the effects of conditioned medium (CM), composed of microvesicles (MVs) and soluble factors present in the supernatant (SN), of bovine endometrial and amniotic cells on embryo quality and rate of blastocyst production. Presumptive zygotes were randomly assigned on Days 1, 3 and 5 after fertilisation to synthetic oviducal fluid with amino acids (SOFaa; control) or to SOFaa supplemented with either 20% endometrial or amniotic CM, 20% SN or 100×106MVsmL-1. Embryos were evaluated on Day 7. For groups supplemented with MVs derived from either endometrial or amniotic cells on Day 1 of culture, blastocysts had developed, but at a lower rate than in the control group. Blastocysts had developed in all groups in which endometrial or amniotic cell-derived CM or MVs were added on Day 3 of culture, but the rate of blastocyst development was significantly lower in both CM groups than in the MVs groups. The addition of all secretome fractions (CM, MVs and SN) derived from either bovine endometrial or amniotic cells on Day 5 of culture resulted in blastocyst production, but only amniotic MVs resulted in a blastocyst production rate comparable to that in the control group. Supplementation of SOFaa on Day 5 resulted in a qualitatively higher number of inner cell mass cells compared with the control group only for the amniotic CM and MVs groups. At day 7, these data were confirmed by RT-qPCR evaluation of genes (Bcl-2-associated X protein (BAX) and glutathione peroxidase 1 (GPX1) involved in apoptosis and protection against reactive oxygen species. In conclusion, of the different secretome fractions tested, only amniotic MVs added to SOFaa resulted in better outcomes than in the control group.

Retinal detachment and convexity intracranial meningioma: an uncommon association.

CASE REPORT: A 70-years-old woman presented with a one week history of progressive loss of vision in the right eye (RE) diagnosed as retinal detachment from 7 to 1 o' clock with retinal break at the 10.30 o' clock associated. Gadolinium enhanced magnetic resonance imaging (MRI) scan of the brain and orbits with fat suppression showed a convexity meningioma. Scleral buckling with a segmental sponge, subretinal fluid drainage and cryopexy were performed with detachment repair. CONCLUSION: In our patient with retinal detachment and convexity meningioma without significant neurological symptoms, performing ocular surgery and close follow up care seemed to be the optimal treatment option.

Spontaneous in-the-bag intraocular lens luxation into the vitreous cavity: last-stage complication of pseudoexfoliative syndrome after phacoemulsification.

PURPOSE: We describe 8 cases of late spontaneous in-the-bag intraocular lens (IOL) luxation into the vitreous cavity, which occurred at the Department of Ophthalmology and Neurosurgery of the University of Siena between January and December 2006. METHODS: In this interventional case series, the medical records of all patients with posterior luxation of in-the-bag IOLs - who had undergone a pars plana vitrectomy with IOL removal and scleral fixation IOL implantation between January and December 2007 at the Department of Ophthalmology and Neurosurgery of Siena, Italy - were retrospectively reviewed. RESULTS: The final post- operative visual acuity was 20/30 or better in 6 patients, while myopic macular degeneration and total retinal detachment limited visual acuity in the remaining 2 patients. CONCLUSION: The high prevalence of pseudoexfoliation (PEX) in the patients who had been operated for cataract phacoemulsification in our department could explain the occurrence of 8 posterior luxations of in-the-bag IOLs in only 1 year. Our study suggests that for the next years we will expect an increase in occurrence of spontaneous in-the- bag IOL luxations in the vitreous cavity. This condition could represent the last stage of PEX syndrome.

The DRPLA CAG repeats in an Italian population sample: evaluation of the polymorphism for forensic applications.

The DRPLA CAG repeats polymorphism has been studied in an Italian population sample. PCR amplification, manual PAGE and silver staining were employed. A total of 16 different alleles, spanning the range from 5 to 21 CAG triplettes, was observed. The heterozygosity was 0.81 and no significant deviation from Hardy-Weinberg equilibrium was found 81 meioses from parentage testing were also analyzed and a Mendelian pattern of inheritance was observed in all cases. In addition, we could successfully type DRPLA locus in some forensic specimens, 1 ng of DNA allowing clear definition of alleles. The authors conclude that the DRPLA CAG repeats analysis may be useful for forensic applications.

Role of computer-assisted analysis of the corneal endothelium in vitreoretinal surgery with intraocular silicone oil injection: a technical report.

The innermost lining of the cornea consists of a single layer of cells called the endothelium. Despite its name, the endothelium of the cornea differs considerably from the vascular endothelium, both functionally and morphologically. The corneal endothelium plays a fundamental role in maintaining the transparency of the corneal membrane, as the result of both its function as a barrier against penetration of the aqueous humor in the parenchyma and its ability to remove water from the stroma (usually referred to as the endothelial "pump" function). Any abnormality in the corneal endothelium causes, first, the impairment of its function as a barrier and pump due to the loss of stromal anti-turgor mechanisms, followed by edema and possible development into keratopathy. The specular microscope is an instrument which makes it possible to see the endothelial "mosaic" in the reflected image of the posterior corneal surface. A large variety of clinical specular microscopes is presently available, both contact and non-contact, which allow, for easy and rapid photography of the corneal endothelium "in vivo". In the present case, we used a non-contact computerized specular microscope to analyze the corneal endothelium in a group of patients affected by retinal detachment who needed to undergo vitreoretinal surgery with immission of silicone oil into the vitreal chamber.

Treatment of high-grade, high-stage prostate cancer with estramustine phosphate or diethylstilbestrol. A double-blind study. The SPCG-1 Study Group. Scandinavian Prostate Cancer Group.

Between 1984 and 1989, 197 patients with T1-4, NX, M1, G2-3 or G3 prostate cancer were randomized to treatment with 560 mg estramustine phosphate (EMP, Estracyt, Emcyt) or 3 mg diethylstilbestrol (DES) per day in a double blind study with stratification on presence or absence of cancer pain at start. A total of 194 patients were evaluated for efficacy of therapy. Time to progression (p = 0.054), to treatment failure (p = 0.036), cancer-specific survival (p = 0.068) as well as overall survival (p = 0.021) were longer in the DES group. There were more patients with prognostic parameters indicating bad prognosis in the EMP group. This trial was designed to study whether EMP had better effect than DES as the primary treatment of high-grade, disseminated prostate cancer. The results did not confirm this hypothesis. On the contrary, treatment with DES had relatively good effect on this very aggressive form of prostate cancer.

Non-conventional ultrasonography (power spectrum analysis) in the management of retinoblastoma.

The differential diagnosis among the diseases that may simulate retinoblastoma is supported by several techniques; however, none of them can give an undeniable answer. In the authors' opinion the tissue characterization by power spectrum analysis of the radiofrequency ultrasound data may play an important role in the backscattered signal spectrum while being sensitive to the spectral shift trend. In order to evaluate the patterns of regression of quiescence of retinoblastoma foci after conservative treatment the authors try to create a model of tissue characterization that provides information not available in conventional A & B scan ultrasonography about cell type, vascularization and necrosis.

The prognostic significance of nuclear DNA content in prostatic carcinoma.

The prognostic significance of ploidy level was studied in prostatic carcinoma and compared with the prognostic significance of morphologic grade and clinical stage. A nonselected group of 145 patients was studied in whom prostatic carcinoma was diagnosed by fine-needle aspiration biopsy at the Karolinska Hospital in 1966. All patients had endocrine therapy and were observed for 23 years or until death. Ploidy level was determined from cytophotometric measurements of Feulgen-stained tumor cell nuclei. The original May-Grünwald-Giemsa-stained cytologic slides, on which the cancer diagnoses were based in 1966, were destained and subsequently Feulgen stained for cytophotometric analysis. From the Feulgen-DNA data, the tumors were classified as near-diploid, near-tetraploid, and highly aneuploid, variants D-type, T-type, and A-type, respectively. The A-type tumors progressed rapidly, and 96% of the patients with this type died of the tumor within 5 years and all patients with this type died within 7 years. D-type and T-type tumors progressed much more slowly. None of the patients with these types of tumors died of the tumor disease within the first 5 years after diagnosis. As many as 12% of the patients (crude survival rate; corresponding to a relative survival rate of 43%) were still alive 15 years after diagnosis. According to multivariate Cox regression analysis, ploidy compared with grade and tumor stage was the strongest predictor of survival.

Uptake of radiocesium by mushrooms.

After the Chernobyl accident in April 1986 the 134Cs, 137Cs and 40K activity of mushrooms of different genera and their corresponding soils from woods located in the province of Umbria (Italy) was determined. The results indicated: a temporally increasing trend for 137Cs up to a "limit value" depending on local fallout in 1986; different cesium concentrations in various mushroom genera; and uptake of potassium from soil that did not appear to compete with cesium uptake. No reliable correlation can be inferred between the cesium content of the soil and those of mushrooms.

Squamous metaplasia in hormonally treated prostatic cancer. Significance during follow-up.

A prospective study of 59 hormonally treated prostatic cancer cases was undertaken during follow-up with sequential transrectal palpation and fine-needle aspiration. At initiation of therapy patients' ages ranged from forty-eight to eighty-two years (median, 68 years). The usual follow-up interval was 6 +/- 3 months, and follow-up periods ranged from six months to one hundred twenty months (median 48 months). The cytologic findings were categorized under four cytologic response types and palpation findings under five response grades. Four different degrees of squamous metaplasia (1+, 2+, 3+, 4+) were observed in smears, depending on semiquantitative determination of squamous metaplastic cells in relation to the total amount of benign and malignant epithelial cells in the smear. There were 341 follow-up observations in which both transrectal fine-needle aspiration cytology and palpation were done. In 306 of these, cytologic findings were found to be adequate. Comparison of squamous metaplasia with cytologic response types revealed a highly significant difference between the benign state and recurrence/frank malignancy. This was also true when frequency of squamous metaplasia was compared with palpatory response grades. It was found that squamous metaplasia can be a valuable adjunct to other cytomorphologic changes such as shrinkage of tumor cell size and decrease in size of nucleoli or its disappearance, in determining response to hormonal therapy.

DNA as a prognostic marker in advanced high-grade prostatic cancer. A preliminary report. SPCG-I study.

This is a presentation of some preliminary data from SPCG-I, a multicenter study started in 1984 by the Scandinavian Prostatic Cancer Group. It is a randomized double-blind study comparing estramustine phosphate and diethylstilbestrol in the primary treatment of 195 patients with T1-4, NX, M1, G2-3 prostatic cancer. The code is not yet broken. This presentation describes the impact of the pretreatment parameters performance status, pain, tumor burden, grade and DNA-ploidy of the prostate tumor, on time to progression and overall survival. DNA studies have so far only been completed in 66 of the 195 patients. For the whole group of 195 patients, pain (p less than 0.004) and tumor grade (p less than 0.02) had the most significant impact on time to progression, and performance status (p less than 0.01) and grade (p less than 0.03) on overall survival. In the small group of 66 patients where the DNA pattern of the primary tumor was evaluated, no parameter had any significant correlation to time to progression and overall survival. This study is still continuing.

Indomethacin modulation of monocyte cytokine release following pelvic irradiation for cancer.

Pelvic irradiation for urogenital cancer reduced monocyte release of tumour necrosis factor alpha (TNF-alpha). Addition of indomethacin to monocyte cultures increased TNF-alpha production after but not before irradiation. E. coli lipopolysaccharide (LPS) increased TNF-alpha release before as well as after radiation therapy and addition of indomethacin to LPS-stimulated monocytes further increased TNF-alpha production following radiotherapy. Spontaneous interleukin-1 (IL-1) release was increased in the cancer patients and was not significantly affected by radiation therapy. LPS increased IL-1 release before as well as after irradiation, but indomethacin did not further change IL-1 secretion. These findings suggest that prostaglandins differentially regulate TNF-alpha and IL-1 release. Administration of cyclo-oxygenase inhibitors during radiation therapy might increase TNF-alpha release in vivo and thereby enhance the host defence against tumours.

Improved early detection of recurrence in prostatic carcinoma following hormonal therapy. Combined use of palpation and fine needle aspiration cytology.

Fifty-nine hormonally treated prostatic carcinoma patients were prospectively followed by rectal examination and fine needle aspiration cytology at six-month intervals for periods ranging from six to one hundred twenty months (median follow-up 48 mos). The cytologic findings and clinical impressions were divided into four or five categories, respectively, ranging from benign to clearly malignant. Cytologic material and clinical descriptions suitable for evaluation were available for 306 follow-up examinations. Of these, 209 were cytologically benign (including 191 without and 18 with clinical evidence of malignancy). The remaining 97 examinations showed cytologic evidence of malignancy of which 50 were clinically apparent (51.5%). While the correlation of clinical and cytologic findings was good (91.4%) in cases with no evidence of disease activity, our findings indicate that many recurrences which are occult to clinical examination may be detected by cytologic examination.

Adjuvant bestatin (Ubenimex) treatment following full-dose local irradiation for bladder carcinoma.

The clinical value of adjuvant bestatin (Ubenimex) immunotherapy has been examined in a group of patients with urinary bladder cancer. Patients with non-metastatic transitional cell carcinoma of the bladder, scheduled for full-dose local irradiation therapy (64 Gy), were randomly allocated to adjuvant oral bestatin treatment (30 mg daily for at least 1 year), starting at completion of irradiation, or no bestatin. The trial included 194 evaluable patients with a follow-up period of 1.5-9.5 years. The overall survival of the two groups of patients did not differ statistically significantly (97 patients in each). Subgrouping of the patient material gave no evidence that the clinical efficacy of bestatin is related to sex, tumor category or malignancy grade.

Recombinant leukocyte interferon alpha-2a and medroxyprogesterone in advanced renal cell carcinoma. A randomized trial.

In a randomized study of advanced renal cell carcinoma 60 patients were allocated to treatment with either recombinant interferon alpha-2a or medroxyprogesterone acetate. Correlation between the dose of interferon alpha-2a and plasma-concentration indicated linear kinetics. Survival was similar in the two treatment groups. Only one complete and one partial response were seen in the interferon group and only one complete response in the medroxyprogesterone group, indicating a low therapeutic potential of both interferon and medroxyprogesterone. Interferon influenced the serum liver enzyme levels; increased transaminases were seen in 17 patients treated with interferon but in only four patients in the medroxyprogesterone group. Two patients had very high serum liver-enzyme levels concomitant with intolerable tiredness, in both patients the symptoms disappeared and the enzymes normalized after discontinuation of the interferon treatment. Antibodies to interferon developed frequently in patients receiving high dose oligomeric interferon therapy but rarely in patients receiving low dose monomeric interferon treatment.

Determination of false negative rate in fine-needle aspiration of prostate. Study of sequential aspirations in 30 untreated carcinoma patients.

Despite several studies comparing the diagnostic accuracy of core needle biopsy and fine-needle aspiration (FNA) in the diagnosis of prostatic adenocarcinoma, no clear picture of the false negative rate for FNA is available. We studied 101 follow-up FNA evaluations in 30 patients with untreated well-differentiated carcinoma. The patients were followed for a median of two years and seven months and had from two to seven FNA follow-up evaluations (median = 2.5 evaluations per patient). If three inadequate evaluations are omitted, the combined rate of positive plus suspicious FNA biopsies was 92.9 percent (88.8% plus 4.1%, respectively). The residual false negative rate was 7.1 percent. The clinical features and follow-up data are described. A semiquantitative definition of "suspicious for malignancy" is suggested. We conclude that in experienced hands. FNA is a highly efficacious method for the diagnosis of prostatic adenocarcinoma.

Circulating LAK-1 cells and autologous mixed lymphocyte reaction in patients with hepatocellular carcinoma.

Autologous mixed lymphocyte reaction (AMLR) and phenotypical composition of circulating lymphocytes obtained from nine subjects with untreated hepatocellular carcinoma (HCC); three HCC patients locally treated by means of intraarterial chemoembolization; six subjects with gut-derived carcinoma (GDC); nine chronic active liver disease patients (CALD), and 14 normal controls have been evaluated, using monoclonal antibodies (MAB) against CD5, CD8, CD4 glycoproteins and anti-LAK-1 molecule, a novel 120-KD surface antigen that is present on the membrane of large granular lymphocytes, by means of classical indirect immunofluorescence technique. Autologous mixed lymphocyte reaction was significantly reduced in all patients, along with CD4-positive cells that are the responder cells in this reaction. A relative increase in the percentage of LAK cells was also observed in neoplastic patients with a normalization after local treatment of the HCC. In the light of promising results obtained by Rosenberg et al. by adoptive transfer of LAK cells activated with Interleukin-2 (IL2) in various cancers, our results support a similar therapeutic trial in HCC patients.

Rectal palpation and transrectal fine needle aspiration of the prostate in the monitoring of prostate cancer: a study of 59 patients during treatment with estramustine phosphate or estrogens.

Fifty-nine hormonally treated prostatic carcinoma patients were prospectively followed by rectal palpation and fine needle aspiration cytology (FNAC) at 6 month intervals for periods ranging from 6 to 120 months (median follow-up, 48 months). The cytologic impressions and palpatory findings were divided into four categories, respectively, ranging from benign to clearly malignant. Cytologic material and palpatory descriptions suitable for evaluation were available for 306 follow-up examinations, with a mean number of follow-up examinations per patient of five (range, 1-13). Tumor relapse was noted in 26 patients and was diagnosed at the same time by FNAC and palpation in six patients. In 16 of the patients tumor progression was first noted by cytology, and in four patients relapse was first detected by rectal palpation. Different patterns of cytologic and palpatory findings during the development of remission and progression of the tumors and drawbacks of the methods are discussed.