Virulent Pseudorabies Virus Infection Induces a Specific and Lethal Systemic Inflammatory Response in Mice.

Pseudorabies virus (PRV) is an alphaherpesvirus that infects the peripheral nervous system (PNS). The natural host of PRV is the swine, but it can infect most mammals, including cattle, rodents, and dogs. In these nonnatural hosts, PRV always causes a severe acute and lethal neuropathy called the "mad itch," which is uncommon in swine. Thus far, the pathophysiological and immunological processes leading to the development of the neuropathic itch and the death of the animal are unclear. Using a footpad inoculation model, we established that mice inoculated with PRV-Becker (virulent strain) develop a severe pruritus in the foot and become moribund at 82 h postinoculation (hpi). We found necrosis and inflammation with a massive neutrophil infiltration only in the footpad and dorsal root ganglia (DRGs) by hematoxylin and eosin staining. PRV load was detected in the foot, PNS, and central nervous system tissues by quantitative reverse transcription-PCR. Infected mice had elevated plasma levels of proinflammatory cytokines (interleukin-6 [IL-6] and granulocyte colony-stimulating factor [G-CSF]) and chemokines (Gro-1 and monocyte chemoattractant protein 1). Significant IL-6 and G-CSF levels were detected in several tissues at 82 hpi. High plasma levels of C-reactive protein confirmed the acute inflammatory response to PRV-Becker infection. Moreover, mice inoculated with PRV-Bartha (attenuated, live vaccine strain) did not develop pruritus at 82 hpi. PRV-Bartha also replicated in the PNS, and the infection spread further in the brain than PRV-Becker. PRV-Bartha infection did not induce the specific and lethal systemic inflammatory response seen with PRV-Becker. Overall, we demonstrated the importance of inflammation in the clinical outcome of PRV infection in mice and provide new insights into the process of PRV-induced neuroinflammation.IMPORTANCE Pseudorabies virus (PRV) is an alphaherpesvirus related to human pathogens such as herpes simplex virus 1 and varicella-zoster virus (VZV). The natural host of PRV is the swine, but it can infect most mammals. In susceptible animals other than pigs, PRV infection always causes a characteristic lethal pruritus known as the "mad itch." The role of the immune response in the clinical outcome of PRV infection is still poorly understood. Here, we show that a systemic host inflammatory response is responsible for the severe pruritus and acute death of mice infected with virulent PRV-Becker but not mice infected with attenuated strain PRV-Bartha. In addition, we identified IL-6 and G-CSF as two main cytokines that play crucial roles in the regulation of this process. Our findings give new insights into neuroinflammatory diseases and strengthen further the similarities between VZV and PRV infections at the level of innate immunity.

Pseudorabies Virus US3 Protein Kinase Protects Infected Cells from NK Cell-Mediated Lysis via Increased Binding of the Inhibitory NK Cell Receptor CD300a.

UNLABELLED: Several reports have indicated that natural killer (NK) cells are of particular importance in the innate response against herpesvirus infections. As a consequence, herpesviruses have developed diverse mechanisms for evading NK cells, although few such mechanisms have been identified for the largest herpesvirus subfamily, the alphaherpesviruses. The antiviral activity of NK cells is regulated by a complex array of interactions between activating/inhibitory receptors on the NK cell surface and the corresponding ligands on the surfaces of virus-infected cells. Here we report that the US3 protein kinase of the alphaherpesvirus pseudorabies virus (PRV) displays previously uncharacterized immune evasion properties: it triggers the binding of the inhibitory NK cell receptor CD300a to the surface of the infected cell, thereby providing increased CD300a-mediated protection of infected cells against NK cell-mediated lysis. US3-mediated CD300a binding was found to depend on aminophospholipid ligands of CD300a and on group I p21-activated kinases. These data identify a novel alphaherpesvirus strategy for evading NK cells and demonstrate, for the first time, a role for CD300a in regulating NK cell activity upon contact with virus-infected target cells. IMPORTANCE: Herpesviruses have developed fascinating mechanisms to evade elimination by key elements of the host immune system, contributing to their ability to cause lifelong infections with recurrent reactivation events. Natural killer (NK) cells are central in the innate antiviral response. Here we report that the US3 protein kinase of the alphaherpesvirus pseudorabies virus displays a previously uncharacterized capacity for evasion of NK cells. Expression of US3 protects infected cells from NK cell-mediated lysis via increased binding of the inhibitory NK cell receptor CD300a. We show that this US3-mediated increase in CD300a binding depends on aminophospholipids and on cellular p21-activated kinases (PAKs). The identification of this novel NK cell evasion strategy may contribute to the design of improved herpesvirus vaccines and may also have significance for other PAK- and CD300a-modulating viruses and cancer cells.

Inter-laboratory evaluation of the ISO standard 11063 "Soil quality - Method to directly extract DNA from soil samples".

Extracting DNA directly from micro-organisms living in soil is a crucial step for the molecular analysis of soil microbial communities. However, the use of a plethora of different soil DNA extraction protocols, each with its own bias, makes accurate data comparison difficult. To overcome this problem, a method for soil DNA extraction was proposed to the International Organization for Standardization (ISO) in 2006. This method was evaluated by 13 independent European laboratories actively participating in national and international ring tests. The reproducibility of the standardized method for molecular analyses was evaluated by comparing the amount of DNA extracted, as well as the abundance and genetic structure of the total bacterial community in the DNA extracted from 12 different soils by the 13 laboratories. High quality DNA was successfully extracted from all 12 soils, despite different physical and chemical characteristics and a range of origins from arable soils, through forests to industrial sites. Quantification of the 16S rRNA gene abundances by real time PCR and analysis of the total bacterial community structure by automated ribosomal intergenic spacer analysis (A-RISA) showed acceptable to good levels of reproducibility. Based on the results of both ring-tests, the method was unanimously approved by the ISO as an international standard method and the normative protocol will now be disseminated within the scientific community. Standardization of a soil DNA extraction method will improve data comparison, facilitating our understanding of soil microbial diversity and soil quality monitoring.

Long-term safety and efficacy of a once-daily formulation of alfuzosin 10 mg in patients with symptomatic benign prostatic hyperplasia: open-label extension study.

OBJECTIVES: To evaluate the long-term safety and efficacy of a new, once-daily (o.d.) prolonged-release formulation of the clinically uroselective alpha1-blocker, alfuzosin, in patients with symptomatic benign prostatic hyperplasia (BPH). METHODS: This is a 9-month open-label extension of a 3-month double-blind, placebo-controlled evaluation of alfuzosin 10 mg o.d. and standard alfuzosin 2.5 mg, three times daily (t.i.d.), administered without dose titration in both cases. A total of 311 patients continued in the extension phase and all received alfuzosin 10 mg o.d. Efficacy was evaluated in all patients enrolled in the extension phase (n = 311). Safety was assessed in all patients exposed to alfuzosin, whether in the double-blind or extension phase (n = 360). RESULTS: Mean international prostate symptom score (IPSS) improved significantly, from 17.1 to 9.3 (P < 0.0001), and mean peak flow rate (PFR) (assessed at through plasma levels) increased significantly, from 9.1 to 11.3 ml/s (P < 0.0001), between baseline (i.e. beginning of the double-blind phase) and the endpoint of the extension phase. Quality of life (QOL) index also improved significantly, from 3.3 to 2.1 (P < 0.0001). Alfuzosin was well tolerated, with only 16 of 360 patients (4.4%) reporting adverse events potentially related to alpha-blockade (mainly dizziness). Ejaculation disorders were infrequent (0.6%) and did not show a relationship to treatment. The incidence of asymptomatic orthostatic hypotension was low (2.8%), and no age effect was identified. CONCLUSIONS: Alfuzosin 10 mg o.d. provides effective relief from BPH, and clinical benefits are maintained up to 12 months. This study also demonstrates the satisfactory long-term safety of this formulation, and its safe use even in at-risk populations.

Efficacy and safety of a new prolonged release formulation of alfuzosin 10 mg once daily versus alfuzosin 2.5 mg thrice daily and placebo in patients with symptomatic benign prostatic hyperplasia. ALFORTI Study Group.

OBJECTIVES: To assess the efficacy and safety of a new prolonged release formulation of the uroselective alpha(1)-blocker alfuzosin for a once-daily dosing regimen in patients with lower urinary tract symptoms (LUTS) suggestive of symptomatic benign prostatic hyperplasia (BPH). METHODS: After a 1-month run-in period, 447 patients were randomly allocated in a double-blind placebo-controlled study to receive alfuzosin 10 mg once daily (n = 143), alfuzosin 2.5 mg thrice daily (n = 150) or placebo (n = 154) for 3 months. At inclusion, 46% of the randomised population had concomitant cardiovascular disease and 30% received an antihypertensive treatment. Uroflowmetry was performed close to trough plasma concentration of alfuzosin once daily to demonstrate the 24-hour coverage with this formulation. RESULTS: Both alfuzosin formulations significantly improved urinary symptoms versus placebo assessed using the International Prostate Symptom Score (alfuzosin 10 mg once daily: -6.9; alfuzosin 2.5 mg thrice daily: -6.4; placebo: -4.9, p = 0.005). Peak flow rate increased significantly with alfuzosin 10 mg once daily (+2.3 ml/s, p = 0.03 vs. placebo) and with alfuzosin 2.5 mg thrice daily (+3.2ml/s, p<0.0001 vs. placebo) compared to placebo (+1.4 ml/s). Overall both formulations of alfuzosin were well tolerated in comparison with placebo. In addition, vasodilatory adverse events appeared to be less frequent with the once daily than the thrice daily formulation (6.3 vs. 9.4%, respectively). No first-day effect was reported with alfuzosin once daily and the effect on blood pressure did not differ from those observed in placebo, both in normotensive and hypertensive patients. No specific sexual dysfunction including ejaculation disorder was reported in the alfuzosin 10 mg once-daily group. CONCLUSION: The new once-daily formulation of alfuzosin administered at a dose of 10 mg daily is an effective 24-hour treatment of LUTS associated with BPH. Alfuzosin is as effective as the immediate formulation and shows a better cardiovascular safety. The better safety profile enables the same dose to be used in all patients, providing the patients with the benefits of a once-daily administration.

[The septic kidney: primary nephrectomy or conservative surgery? A clinical study of 110 cases (author's transl)]. / Die septische Niere: Nephrektomie oder Organerhaltung? Eine klinische Studie an 110 Fällen.

The "Septic Kidney" usually originates from infected hydronephrosis. The clinical appearance is characterized by poor general condition, in particular shock, severe flank pain, high fever with chills, leucocytosis and often azotemia. The pathogenesis and definition are discussed on the basis of 110 cases. The various therapeutic modalities such as primary nephrectomy, conservative surgery or endoscopic instrumentation are compared and the indications defined. Conservative procedures are preferred over primary nephrectomy.

Spontaneous phasic activity of the detrusor: a cause of uninhibited contractions in unstable bladders?

Uninhibited detrusor contractions of neurogenic origin have been described repeatedly. Since a spontaneous myogenic activity of the detrusor has been demonstrated in isolated muscle strips, it seems reasonable that an increase of this spontaneous contraction activity may induce bladder instability of pure myogenic origin. 30 patients separated into two groups were investigated. All patients suffered from involuntary contractions of the detrusor, accompanied by loss of urine. Nifedipin was applied as a selective Ca-antagonistic drug to block the phasic detrusor activity. In our experiment, involuntary detrusor contractions could not be suppressed, and also bladder capacity could not be increased by this drug. This result leads to the assumption that the spontaneous phasic activity of the detrusor does not induce any involuntary bladder contraction.

Paranephric abscess: a changing concept.

The aetiology of the paranephric abscess has been greatly modified since the introductiion of antibiotics. Its frequency has decreased but its prognosis has not been improved. 20 cases of paranephric abscess have been studied retrospectively. Its main cause is no longer the haematogenic staphylococcus infection but primary kidney infection. The disease is difficult to diagnose as the evolution is slow and the symptoms unspecific. The main symptoms are: abdominal pain, feeling of physical prostation, subfebrility, acute pain in the flank and significant increase in blood sedimentation rate. The infection is due in most cases to a silent kidney or a kidney stone with pyonephrosis. Accompaying diabetes mellitus was often observed.

[Pseudotumor of the renal pelvis during nonspecific inflammatory granuloma (author's transl)]. / Pseudotumor des Nierenbeckens durch ein unspezifisch-entzündliches Granulom.

Differential diagnosis of tumors of the renal pelvis cannot be assured preoperatively in every case. A 69-year-old man is reported who had an inflammatory granuloma of the renal pelvis after perforation of an unspecific abscess. The granuloma appeared radiologically like an advanced tumor of the renal pelvis.

Effects of beta-adrenergic stimulating and blocking agents on the dynamics of the human bladder outlet.

The alpha-adrenergic innervation of the functional urethra is a well-known fact, while beta-adrenergic influence is rather unknown until now. We studied the influence of beta-stimulating and beta-blocking agents on the human urethra by the urethral pressure profile (UPP). A decrease of the UPP under orciprenaline sulfate and an increase under propranolol could be mentioned.

[Vesicoureterorenal reflux: an inherited disease? (author's transl)]. / Der vesiko-uretero-renale Reflux: ein vererbbares Leiden

A family case history is presented with an increased incidence of vesicorenal reflux: four sons had a reflux while a daughter and the parents were healthy in this respect. From the literature and on own studies the possibility of reflux as an inherited disease is discussed. A new finding is the coincidence of reflux and the histocompatibility-antigen HLA-A2.