Androgen-regulated miR-32 targets BTG2 and is overexpressed in castration-resistant prostate cancer.

The androgen receptor (AR) signaling pathway is involved in the emergence of castration-resistant prostate cancer (CRPC). Here, we identified several androgen-regulated microRNAs (miRNAs) that may contribute to the development of CRPC. Seven miRNAs, miR-21, miR-32, miR-99a, miR-99b, miR-148a, miR-221 and miR-590-5p, were found to be differentially expressed in CRPC compared with benign prostate hyperplasia (BPH) according to microarray analyses. Significant growth advantage for LNCaP cells transfected with pre-miR-32 and pre-miR-148a was found. miR-32 was demonstrated to reduce apoptosis, whereas miR-148a enhanced proliferation. Androgen regulation of miR-32 and miR-148a was confirmed by androgen stimulation of the LNCaP cells followed by expression analyses. The AR-binding sites in proximity of these miRNAs were demonstrated with chromatin immunoprecipitation (ChIP). To identify target genes for the miRNAs, mRNA microarray analyses were performed with LNCaP cells transfected with pre-miR-32 and pre-miR-148a. Expression of BTG2 and PIK3IP1 was reduced in the cells transfected with pre-miR-32 and pre-miR-148a, respectively. Also, the protein expression was reduced according to western blot analysis. BTG2 and PIK3IP1 were confirmed to be targets by 3'UTR-luciferase assays. Finally, immunostainings showed a statistically significant (P<0.0001) reduction of BTG2 protein in CRPCs compared with untreated prostate cancer (PC). The lack of BTG2 staining was also associated (P<0.01) with a short progression-free time in patients who underwent prostatectomy. In conclusion, androgen-regulated miR-32 is overexpressed in CRPC, leading to reduced expression of BTG2. Thus, miR-32 is a potential marker for aggressive disease and is a putative drug target in PC.

Diagnostic and prognostic signatures from the small non-coding RNA transcriptome in prostate cancer.

Prostate cancer (PCa) is the most frequent male malignancy and the second most common cause of cancer-related death in Western countries. Current clinical and pathological methods are limited in the prediction of postoperative outcome. It is becoming increasingly evident that small non-coding RNA (ncRNA) species are associated with the development and progression of this malignancy. To assess the diversity and abundance of small ncRNAs in PCa, we analyzed the composition of the entire small transcriptome by Illumina/Solexa deep sequencing. We further analyzed the microRNA (miRNA) expression signatures of 102 fresh-frozen patient samples during PCa progression by miRNA microarrays. Both platforms were cross-validated by quantitative reverse transcriptase-PCR. Besides the altered expression of several miRNAs, our deep sequencing analyses revealed strong differential expression of small nucleolar RNAs (snoRNAs) and transfer RNAs (tRNAs). From microarray analysis, we derived a miRNA diagnostic classifier that accurately distinguishes normal from cancer samples. Furthermore, we were able to construct a PCa prognostic predictor that independently forecasts postoperative outcome. Importantly, the majority of miRNAs included in the predictor also exhibit high sequence counts and concordant differential expression in Illumina PCa samples, supported by quantitative reverse transcriptase-PCR. Our findings provide miRNA expression signatures that may serve as an accurate tool for the diagnosis and prognosis of PCa.

Association of different HLA antigens with various toxic effects of gold salts in rheumatoid arthritis.

An association of gold induced proteinuria with HLA-D(R)3 has been reported. To investigate other possible relationships between gold toxicity and HLA antigens we studied 85 patients with rheumatoid arthritis (RA) divided into four subgroups: patients with gold induced interstitial pneumonitis, mucocutaneous lesions, proteinuria, and patients without gold toxicity. The HLA frequencies in patient groups and 283 healthy controls were compared in different pairwise combinations. Gold induced pneumonitis was associated with HLA-B40 and Dw1. An association between gold induced proteinuria and HLA-Dw3 was also seen. The increased prevalence of Dw4 in RA was observed only in the control patient group without gold induced side effects. The frequencies of HLA-B7 and Dw2 were decreased in all patient groups compared with the control population. These results further support the view of the heterogeneity of RA as manifested by the unique HLA associations with resistance and susceptibility to gold induced side effects characterising different subgroups.

Rheumatoid arthritis with pleural effusion includes a subgroup with autoimmune features and HLA-B8, Dw3 association.

Twenty-eight patients with rheumatoid pleural effusion were investigated to examine the frequency of HLA antigens as compared with 56 rheumatoid arthritis (RA) patients without this intrathoracic manifestation of RA and with 283 healthy controls. HLA-B8 was strongly associated with the presence of pleural effusion (PE) in RA patients. A high prevalence (71%) of B8/Dw3 was found among male RA patients of the PE group in whom the joint disease had begun at an age over 50 years and who also had besides pleuritis other intrathoracic manifestations of RA associated with high rheumatoid factor titres and low complement (C4) levels in sera. Actually, the HLA-B8 association was not seen in the rest of the PE group. The finding may be related to the heterogeneity of RA, a male subgroup of the disease being characterized by multiple intrathoracic manifestations and genetically associated with the large group of autoimmune disorders, such as SLE, characterized by high prevalences of HLA-B8 and D(R)3.

Xanthoma tendinosum in a normolipemic ectodermal dysplasia patient.

A female patient with congenital ectodermal dysplasia is described, who developed xanthoma tendinosum in her hands simulating rheumatoid arthritis. The serum lipids of this patient were normal. Both ectodermal dysplasia and xanthoma tendinosum in a normolipemic patient are rather uncommon and to our knowledge have not been previously reported in one and the same patient.

Keratodermia blenorrhagica, arthritis, and polymyositis with cardiopulmonary complications.

A patient is described who had polymyositis with arthritis, keratodermia blenorrhagica, pulmonary fibrosis, and cardiac failure with a right bundle branch block. The cutaneous lesions on his palms and soles, considered to be specific for Reiter's syndrome, pointed to an overlapping of polymyositis with features of this syndrome. Findings typical of myositis were present. In addition a muscle biopsy showed sarcolemmal and endomysial deposits of IgG and IgM, and the serum level of IgM was elevated. Because skin lesions appeared at the height of symptoms in other organs, the possibility is raised that both might stem from a common immune process.

Concentration of carprofen in the serum and the synovial fluid in rheumatoid arthritis patients.

Concentrations of carprofen in the serum and the synovial fluid were determined in 13 patients with classical or confirmed rheumatoid arthritis. Four hours after the oral administration of 100 mg of carprofen, as one capsule, concentrations in the synovial fluid were found to be between 3.5 micrograms/ml and 9.7 micrograms/ml, with a mean value (+/- SD) of 6.2 +/- 1.4 micrograms/ml, and concentrations in the serum were between 4.7 micrograms/ml and 14.8 micrograms/ml, with a mean value of 9.8 +/- 2.9 micrograms/ml. The carprofen concentrations in synovial fluid varied from 36.3 to 103.0% of those in serum, with a mean value of 65.0 +/- 14.4%. There is a highly significant correlation between carprofen concentrations in the serum and the synovial fluid.

A double blind crossover study of carprofen and indomethacin in patients with rheumatoid arthritis.

Twenty-eight patients with definite or classic rheumatoid arthritis were studied in a four-month double blind crossover trial. Patients were randomly divided into two groups, and received either carprofen, 300 mg/day, or indomethacin, 75 mg/day, for a period of two months, and then the other medication for a further two months. The classic procedures and measurements for evaluation of activity of rheumatoid arthritis were used. Laboratory tests were also carried out to assess toxicity. Both drugs were effective for the long term treatment of rheumatoid arthritis, and there were no statistically significant differences between the carprofen and indomethacin periods, or between groups. Although the incidence of adverse reactions was similar for both drugs, cutaneous symptoms were more frequent with carprofen than with indomethacin, whereas gastrointestinal and central nervous system reactions were found more often with indomethacin.

Treatment of finger joints with local steroids. A double-blind study.

The study was performed in 24 patients all having rheumatoid arthritis affecting the finger joints. Randomly, and in a double-blind fashion, the inflamed interphalangeal and proximal interphalangeal joints of one hand were injected with either triamcinolone hexacetoide (TH) or methylprednisolone (MP) and those of the other hand vice versa. A total of 120 affected swollen finger joints were injected--59 joints with TH and 61 with MP. The duration of effect and the possible side effects were followed up for a period of 6 months. All injections produced clinically significant effects. There were no significant differences between the two treatment groups at the start of the treatment, but after 6 months the results in the TH group were significantly better. However, there were also more joints with skin and soft tissue atrophy in this group than in the MP group.

The effect of rheumatoid synovial fluid macrophages on DNA, glycosaminoglycan and collagen synthesis by synovial fibroblasts.

The effects of soluble factors secreted by peripheral blood monocytes and rheumatoid synovial fluid macrophages were tested on human synovial fibroblast cultures. Both monocytes and macrophages liberated factors which reduced DNA synthesis (3H-thymidine incorporation) by synovial fibroblasts. Monocyte and macrophage factors stimulated hyaluronic acid synthesis. The activation obtained with rheumatoid synovial macrophages was considerably greater than that with monocytes. Foetal bovine serum was found to have a clear stimulatory effect on the synthesis of collagen and other proteins by fibroblasts. The effects of monocyte and macrophage factors on protein synthesis in synovial fibroblasts were small: collagen synthesis was slightly increased relative to other extracellular proteins.

Healing of erosions in rheumatoid arthritis.

In rheumatoid arthritis (RA) such early radiographic changes as soft tissue swelling and osteoporosis may well be reversible. Narrowing of the joint space and bone erosion are late changes generally considered to be non-reversible. Our observation suggest, however, that healing of bone erosions, especially in joints in which inflammation had disappeared, is more common than is generally believed. We describe three patients with classical RA in whom serial radiographs showed that the bone erosion in one or more of the joints of their hands or feet had healed.

Triamcinolone hexacetonide in the treatment of therapy-refractory gonitis.

Thirty-seven patients suffering from sustained gonitis with recurrent hydrops, not satisfactorily controlled by systemic treatment, participated in the study. Twenty-three patients had bilateral and fourteen patients had unilateral gonitis. controls were obtained through the use of the contra-lateral knee joint in the twenty-three patients with bilateral gonitis. A total of 60 knees were randomized into three groups of 20 knees and received either 30 mg triamcinolone hexacetonide (TH), 60 mg methylprednisolone (MP) or 9 mg betamethasone (BM). The clinical parameters evaluated were: recurrence of hydrops, pain, local tenderness, range of joint movement and joint circumference. The period of time elapsing between the injection and the need for a new injection or other treatment was used as the criterion for the duration of effect. Relief lasting for an average of 176 days was obtained in the 20 knees treated with local injections of TH, compared to 125 days for MP and 79 days for BM. No general or local adverse effects were noted in the patients studied. The observed long duration of effect and the absence of adverse effects justify the use of TH in cases of therapy-refractory gonitis.

Indoprofen versus indomethacin and placebo. A double-blind three-way cross-over trial in rheumatoid arthritis.

Thirty in-patients suffering from classical or definite R.A. were given indoprofen 200 mg q.i.d., indomethacin 25 mg q.i.d. and placebo, according to a 6-line completely balanced block design. Each patient received the three treatments for ten days each. Patients were examined at baseline and after each treatment cycle. Both active drugs were significantly better than placebo on relief of pain, Ritchie index, and number of active, painful and swollen joints, morning stiffness; only indoprofen significantly improved grip strength. No significant differences were found between the two active drugs. The three treatments were well tolerated.

Synovial fluid lipids in rheumatoid arthritis.

Synovial fluid from the inflamed knee joints of 30 patients with rheumatoid arthritis (RA) contained 2.75 +/- 0.81 mmol/l cholesterol (51% of the corresponding mean serum level) and 0.32 +/- 0.13 mmol/l triglycerides (35% of the corresponding mean serum level). The ratio of synovial fluid triglycerides to cholesterol indicates that in RA the concentration of very low density lipoproteins (VLDL) is lower in synovial fluid than in serum. In a few patients, however, the "relative triglyceride concentration" in synovial fluid (i.e. synovial fluid tg/chol:serum tg/chol) was greater than 1. This concentration did not correlate, moreover, with the clinical activity of the disease.

Pulmonary infiltrates associated with gold therapy. Report of a case.

A case report is presented concerning diffuse pulmonary infiltrates due to sodium aurothiomalate therapy in a 67-year-old woman suffering from rheumatoid arthritis. After the gold treatment was stopped, prednisolone therapy produced complete remission of the pulmonary reaction.

Reversible renal side effects of intra-articular osmic acid injection.

In 101 patients treated for knee joint hydrops with intra-articular osmic acid injection, an increased incidence of proteinuria and microscopic haematuria was noted on the first day after injection. This increase subsided rapidly and had disappeared 3 days after the injection. Three patients showed transient glycosuria. No signs of permanent renal damage appeared in any patient.