Challenges and potential for RNA nanoparticles (RNPs).

The current research of our group focuses on delivery of oligonucleotides and gene vaccines targeted against infectious disease and cancer. Recently we reported a strategy for associating DNA to gold via protamine delivery enhancing material which here we suggest might be applicable to RNA and to other nanoparticles. An important new modality for such RNA based nanoparticles (RNPs) is the myriad of genes now known to undergo alternative splicing. Here we will review some important issues for the binding, stabilization and delivery of RNA, particularly splice-site switching oligomers (SSOs) via these RNPs in order to achieve selective molecular therapeutic effects and unlock their potential as chemotherapeutic agents.

Indications for penile revascularization and long-term results.

The purpose of this study was to analyse long-term results of penile revascularization using Hauri's method in 124 patients with a mean follow-up of 54 months. Of 176 patients undergoing this procedure, 124 were available for detailed analysis. The patients ranged in age from 22 to 71. A total of 25.8% of the patients (32/124) responded to intracavernous injection (ICI). Postoperatively, 74 patients (59.7%) exhibited spontaneous erections. Patients were classified as 'satisfied' or 'dissatisfied'. In those who were satisfied, a high correlation was found (63/74 = 85%) between graft patency, as judged by ultrasound, and erectile function. The benefit for non-responders to ICI (60/92) was higher than for responders (14/32). Only five of 12 diabetics profited from penile revascularization. A serious complication was glans hyperemia in 9/124 cases (7%). Based on this experience, the following indicators are recommended for case selection: (i) non-responder to ICI; (ii) age less than 55 years; (iii) nondiabetic; (iv) cavernous leakage excluded; (v) stenosis in the internal pudendal artery.

[Microsurgical testicular denervation as therapy option in chronic testalgia]. / Die mikrochirurgische testikulre Denervierung als Therapieoption der chronischen Testalgie.

Chronic testicular pain represents a challenging urological chronic pain syndrome in terms of adequate diagnosis and therapy. Reported success rates of 55-73% and 10-40% of conservative and surgical interventions are extremely low. We report on microsurgical testicular denervation as therapeutic option in patients with chronic testicular pain (CTP). 12 consecutive patients with CTP were included in our study. After complete diagnostic workup and positive response to testicular nerve blockade, all patients underwent surgery: the cremasteric muscle was dissected by electrocautery, the periadventitial layer of the testicular artery was dissected over a length of 2-3 cm. After a median follow-up of 20.6 months (4-62) 11/12 patients (92%) are pain free. None of the patients suffered from intraa- or postoperative complications. Based on our experience microsurgical testicular denervation should be performed in patients with CTP and no underlying organic disease. However, the high success rate of our surgical procedure can only be maintained if the selection of suitable patients is performed very carefully and a specific organic origin of CTP has been excluded prior to surgery.

Novel non-nucleoside inhibitors of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. 4. 2-Substituted dipyridodiazepinones as potent inhibitors of both wild-type and cysteine-181 HIV-1 reverse transcriptase enzymes.

The major cause of viral resistance to the potent human immunodeficiency virus type 1 reverse transcriptase (RT) inhibitor nevirapine is the mutation substituting cysteine for tyrosine-181 in RT (Y181C RT). An evaluation, against Y181C RT, of previously described analogs of nevirapine revealed that the 2-chlorodipyridodiazepinone 16 is an effective inhibitor of this mutant enzyme. The detailed examination of the structure-activity relationship of 2-substituted dipyridodiazepinones presented below shows that combined activity against the wild-type and Y181C enzymes is achieved with aryl substituents at the 2-position of the tricyclic ring system. In addition, the substitution pattern at C-4, N-5, and N-11 of the dipyridodiazepinone ring system optimum for inhibition of both wild-type and Y181C RT is no longer the 4-methyl-11-cyclopropyl substitution preferred against the wild-type enzyme but rather the 5-methyl-11-ethyl (or 11-cyclopropyl) pattern. The more potent 2-substituted dipyridodiazepinones were evaluated against mutant RT enzymes (L100I RT, K103N RT, P236L RT, and E138K RT) that confer resistance to other non-nucleoside RT inhibitors, and compounds 42, 62, and 67, with pyrrolyl, aminophenyl, and aminopyridyl substituents, respectively, at the 2-position, were found to be effective inhibitors of these mutant enzymes also.

Inhibition of human leukocyte elastase (HLE) by N-substituted peptidyl trifluoromethyl ketones.

A series of tripeptides possessing trifluoromethyl or aryl ketone residues at P1 were prepared and evaluated both in vitro and in vivo as potential inhibitors of human leukocyte elastase (HLE). Tripeptides containing non naturally occurring N-substituted glycine residues at the P2-position have been demonstrated to be potent in vitro inhibitors of HLE, with IC50 values in the submicromolar range. Sterically demanding substituents on the P2-nitrogen have no detrimental effect on in vitro potency. The inhibition process presumably acts via hemiketal formation with the active site Ser195 of HLE, and is facilitated by the strongly electron withdrawing trifluoromethyl functionality. Deletion of the amino acid at the P3-subsite region affords inactive compounds. Valine is the preferred residue at the P1-position, whereas the corresponding glycine, alanine, alpha,alpha-dimethylglycine, or phenylalanine analogues are all inactive. The compounds described herein all confer a high degree of in vitro specificity when tested against representative cysteine, aspartyl, metallo, and other serine proteases. One of the most potent in vitro inhibitors is (3RS)-N-[4-[[[(4-chlorophenyl)sulfonyl]amino]carbonyl]phenyl] oxomethyl]-L-valyl-N-(2,3-dihydro-1H-inden-2-yl)glycine N-[3-(1,1,1-trifluoro-4-methyl-2-oxopentyl)]amide (20i; BI-RA-260) (IC50 = 0.084 microM). Compound 20i was also tested in hamsters in an elastase-induced pulmonary hemorrhage (EPH) model. In this model, intratracheal (it.) administration of 20i, 5 min prior to HLE challenge, effectively inhibited hemorrhage in a dose-dependent manner with an ED50 of 4.8 micrograms. The inhibitor 20i, 20 micrograms administered it. 24, 48, and 72 h prior to HLE challenge, exhibits significant inhibition against hemorrhage at all time points (97%, 64% and 49%, respectively). In a 21-day chronic model of emphysema in hamsters, 200 micrograms of HLE administered it. caused an elastase-induced emphysema in the lungs which can be quantitated histologically utilizing image analysis. In this assay, 20i significantly inhibited pulmonary lesions associated with septal destruction and increased alveolar spaces, when dosed at 20 micrograms it. 5 min prior to challenge with HLE.

Novel non-nucleoside inhibitors of HIV-1 reverse transcriptase. 1. Tricyclic pyridobenzo- and dipyridodiazepinones.

Novel pyrido[2,3-b][1,4]benzodiazepinones (I), pyrido[2,3-b][1,5]benzodiazepinones (II), and dipyrido[3,2-b:2',3'-e][1,4]diazepinones (III) were found to inhibit human immunodeficiency virus type 1 (HIV-1) reverse transcriptase in vitro at concentrations as low as 35 nM. In all three series, small substituents (e.g., methyl, ethyl, acetyl) are preferred at the lactam nitrogen, whereas slightly larger alkyl moieties (e.g., ethyl, cyclopropyl) are favored at the other (N-11) diazepinone nitrogen. In general, lipophilic substituents are preferred on the A ring, whereas substitution on the C ring generally reduces potency relative to the corresponding compounds with no substituents on the aromatic rings. Maximum potency is achieved with methyl substitution at the position ortho to the lactam nitrogen atom; however, in this case an unsubstituted lactam nitrogen is preferred. Additional substituents on the A ring can be readily tolerated. The dipyridodiazepinone derivative 11-cyclopropyl-5,11-dihydro-4-methyl-6H-dipyrido[3,2-b:2',3'-e] [1,4]diazepin-6-one (96, nevirapine) is a potent (IC50 = 84 nM) and and selective non-nucleoside inhibitor of HIV-1 reverse transcriptase, and has been chosen for clinical evaluation.

Inhibition of human immunodeficiency virus type 1 (HIV-1) replication by the dipyridodiazepinone BI-RG-587.

The dipyridodiazepinone human immunodeficiency virus type 1 (HIV-1)-specific reverse transcriptase (RT) inhibitor BI-RG-587 was tested for its ability to inhibit HIV-1 replication in both acutely and chronically infected cell lines. The ability of BI-RG-587 to inhibit steps in the virus replicative cycle other than reverse transcription was also assessed. BI-RG-587 was found to be a potent inhibitor of HIV-1 replication in acutely infected cells (50% inhibitory concentration [IC50] = 37.2 nM), and the sensitivity and kinetics of that inhibition was similar to the known RT inhibitor zidovudine (AZT). Even at 100x IC50, BI-RG-587 had no effect on gp120/CD4 interaction, syncytia formation, or envelope glycoprotein processing. In addition, no inhibition of viral replication or protein production was noted in a chronically infected cell line that produces viral products in an RT-independent manner. Finally, no inhibition of acute HIV-2 replication was noted, even with very high (2500x IC50 for HIV-1) concentrations of BI-RG-587. These results demonstrate that BI-RG-587 is a potent inhibitor of HIV-1 replication and that this inhibition occurs at the point of reverse transcription.

Inhibition of HIV-1 replication by a nonnucleoside reverse transcriptase inhibitor.

A series of dipyridodiazepinones have been shown to be potent inhibitors of human immunodeficiency virus-1 (HIV-1) reverse transcriptase (RT). One compound, BI-RG-587, had a Ki of 200 nanomolar for inhibition of HIV-1 RT that was noncompetitive with respect to deoxyguanosine triphosphate. BI-RG-587 was specific for HIV-1 RT, having no effect on feline and simian RT or any mammalian DNA polymerases. BI-RG-587 inhibited HIV-1 replication in vitro as demonstrated by in situ hybridization, inhibition of protein p24 production, and the lack of syncytia formation in cultured human T cell lines and freshly isolated human peripheral blood lymphocytes. Cytotoxicity studies of BI-RG-587 on human cells showed a high therapeutic index (greater than 8000) in culture.

Calf hematoma mimicking thrombophlebitis: sonographic and computed tomographic appearance.

Five patients with calf hematomas presented with signs and symptoms suggesting thrombophlebitis, obscuring the correct diagnosis. Venography showed no venous thrombosis; further diagnostic studies using ultrasound and/or computed tomography provided the correct diagnosis in all patients. Ultrasound showed a hypoechoic mass clearly demarcated from surrounding soft tissue, while computed tomography showed a well-defined mass whose density depended on the age of the hematoma.

Trauma of occult hydronephrotic kidney.

The occult unilateral hydronephrotic kidney is often discovered during the genitourinary evaluation of patients sustaining blunt abdominal trauma. Few cases have been reported documenting the angiographic, computerized tomography (CT), and ultrasound appearances. Two cases are described which demonstrate that relatively minor trauma can precipitate hematuria and hypovolemic shock. Angiography demonstrated the bleeding site in both cases and was utilized in conjunction with other parameters of clinical assessment to plan initial management. CT and ultrasound proved to be useful noninvasive diagnostic parameters for baseline and follow-up studies in patients undergoing conservative management. They accurately demonstrated the degree of hydronephrosis, residual renal parenchymal, and resolving hematoma.

[Cholesterol in the blood and bile]. / Cholesterin in Blut und Galle.

In a prospective study serum and bile cholesterol concentrations were examined in 233 patients with gallstones. The objective of the study was the selection of patients in which conservative dissolution of gallstones might be successful. The average concentration of cholesterol in serum was 227,46 +/- 41,64 mg/100 ml, in gallbladder bile 8,83 +/- 5,66 mg/100 ml. The cholesterol contents of gallstones was 73,47 +/- 19,47 per cent. Statistical relations were found neither between the cholesterol concentrations in serum and bile nor between the cholesterol contents of gallstones and the cholesterol concentrations in serum and bile. The postoperative cholesterol concentration in serum was remarkably diminished as compared with preoperative values in all patients. A statistically significant decrease of serum cholesterol was seen in patients with histologically proven fatty liver disease. Measuring the cholesterol concentration in serum is no appropriate way for the selection of patients with gallstones for a litholytic treatment. It should be evaluated whether a concomitant fatty liver disease is an indication for a therapeutic trial with chenodesoxycholic acid.

[Microbiological and histologic correlations in inflammatory gallbladder disease (author's transl)]. / Beziehungen zwischen bakteriologischen und histologischen Befunden bei entzündlichen Gallenblasenerkrankungen.

In a hospital population of 243 patients undergoing surgery for gallstones, inflammatory alterations were seen in 177 cases. The search for bacteria was positive in 33 cases. Thus, more than 80 per cent of the cholecystitis cases were abacterial. Under the age of 50, micro-organisms were found in 4.5 per cent only, and in 37 per cent of patients above fifty. In 11 cases of acute cholecystits no bacteria were found at all. Antibiotic treatment is justified in high-risk patients.