On the Reliability of Suction Measurements for Skin Characterization.

in vivo skin characterization methods were shown to be useful in the detection of microstructural alterations of the dermis due to skin diseases. Specifically, the diagnostic potential of skin suction has been widely explored, yet measurement uncertainties prevented so far its application in clinical assessment. In this work, we analyze specific factors influencing the reliability of suction measurements. We recently proposed a novel suction device, called Nimble, addressing the limitations of existing instruments, and applied it in clinical trials quantifying mechanical differences between healthy skin and scars. Measurements were performed with the commercial device Cutometer and with the new device. A set of new suction measurements was carried out on scar tissue and healthy skin, and FE-based inverse analysis was applied to determine corresponding parameters of a hyperelastic-viscoelastic material model. FE simulations were used to rationalize differences between suction protocols and to analyze specific factors influencing the measurement procedure. Tissue stiffness obtained from Cutometer measurements was significantly higher compared to the one from Nimble measurements, which was shown to be associated with the higher deformation levels in the Cutometer and the nonlinear mechanical response of skin. The effect of the contact force exerted on skin during suction measurements was quantified, along with an analysis of the effectiveness of a corresponding correction procedure. Parametric studies demonstrated the inherently higher sensitivity of displacement- over load-controlled suction measurements, thus rationalizing the superior ability of the Nimble to distinguish between tissues.

Reporting of Safety Events during Anti-VEGF Treatment: Pharmacovigilance in a Noninterventional Trial.

AIM: The prospective, noninterventional OCEAN study assessed the safety of intravitreal ranibizumab injections for treatment of neovascular age-related macular degeneration, diabetic macular edema, and retinal vein occlusion under real-world conditions in Germany. METHODS: Adults receiving ≥1 ranibizumab (0.5 mg) injections were recruited by 369 ophthalmologists and followed for 24 months. Information on adverse events (AEs) was reported by the treating physician or detected by the data management team. Collected information included observed AE, AE start and end date, intensity, causal relationship, outcome, severity, suspected drug, and actions taken. RESULTS: 2,687 AEs were reported for 1,176 of the 5,781 patients who had received a total of 32,621 injections: 27.4% nonserious AEs, 30.3% serious AEs, 27.3% nonserious adverse drug reactions (ADRs), and 15.0% serious ADRs. Most patients reported no AEs (79.7%) or only 1 AE (10.3%). AEs were most commonly reported in the Medical Dictionary for Regulatory Activities (MedDRA) System Organ Class (SOC) Eye disorders (9.4% of patients) and General disorders and administration site conditions (5.8%). The most frequent AEs by MedDRA preferred term (PT) were visual acuity reduced (3.5% of patients), intraocular pressure increased (2.5%), and drug ineffective (2.1%). AEs occurred most frequently after 3 or 4 injections (1,129 of 2,687 AEs). The proportion of AEs in the SOC Eye disorders decreased slightly with increasing number of injections, from 39.8% of events after 1 or 2 injections to 29.1% after 5 or more injections. Rates of the most frequently reported PTs did not show any consistent increase with increasing number of injections. A decrease in overall AE rates was observed over the study course. CONCLUSIONS: The results did not raise any new safety concerns for ranibizumab. The findings allow conclusions to be drawn on how pharmacovigilance data can be collected even more effectively in real-world studies to facilitate discussion on benefit-risk ratio.

Two-year-old children but not domestic dogs understand communicative intentions without language, gestures, or gaze.

Infants can see someone pointing to one of two buckets and infer that the toy they are seeking is hidden inside. Great apes do not succeed in this task, but, surprisingly, domestic dogs do. However, whether children and dogs understand these communicative acts in the same way is not yet known. To test this possibility, an experimenter did not point, look, or extend any part of her body towards either bucket, but instead lifted and shook one via a centrally pulled rope. She did this either intentionally or accidentally, and did or did not address her act to the subject using ostensive cues. Young 2-year-old children but not dogs understood the experimenter's act in intentional conditions. While ostensive pulling of the rope made no difference to children's success, it actually hindered dogs' performance. We conclude that while human children may be capable of inferring communicative intent from a wide variety actions, so long as these actions are performed intentionally, dogs are likely to be less flexible in this respect. Their understanding of communicative intention may be more dependent upon bodily markers of communicative intent, including gaze, orientation, extended limbs, and vocalizations. This may be because humans have come under selective pressure to develop skills for communicating with absent interlocutors - where bodily co-presence is not possible.

The ubiquitin ligase Praja1 reduces NRAGE expression and inhibits neuronal differentiation of PC12 cells.

Evidence suggests that regulated ubiquitination of proteins plays a critical role in the development and plasticity of the central nervous system. We have previously identified the ubiquitin ligase Praja1 as a gene product induced during fear memory consolidation. However, the neuronal function of this enzyme still needs to be clarified. Here, we investigate its involvement in the nerve growth factor (NGF)-induced differentiation of rat pheochromocytoma (PC12) cells. Praja1 co-localizes with cytoskeleton components and the neurotrophin receptor interacting MAGE homologue (NRAGE). We observed an enhanced expression of Praja1 after 3 days of NGF treatment and a suppression of neurite formation upon Praja1 overexpression in stably transfected PC12 cell lines, which was associated with a proteasome-dependent reduction of NRAGE levels. Our data suggest that Praja1, through ubiquitination and degradation of NRAGE, inhibits neuronal differentiation. The two murine isoforms, Praja1.1 and Praja1.2, appear to be functionally homologous in this respect.

In utero transmission of porcine torque teno viruses.

Sera and selected tissue homogenates collected from gnotobiotic swine never exposed to the environment or other swine tissues were tested for the presence of porcine torque teno virus (TTV) DNAs by nested and non-nested polymerase chain reactions (PCR) using primers specific for the untranslated region of porcine genogroups (g) 1 and 2. Twenty-three of 105 (21.9%) gnotobiotic piglets were g1- and/or g2-TTV DNA positive. Twenty-three of 27 (85.2%) sow sera, collected at the time of Caesarian derivation of the litters contained either or both TTV genogroup DNAs. These data demonstrate that porcine TTV may be transmitted to piglets by the in utero route and that the incidence of fetal infection is high.

Gene expression of the human Torque Teno Virus isolate P/1C1.

Torque Teno Virus (TTV) has been assigned to the floating genus Anellovirus. TTV ssDNA genomes have a size of 3.6 to 3.8 kb and display up to 30% nucleotide diversity. The pathogenic potential of TTV is under investigation. To address a putative link of pathogenicity with the observed sequence variations, the transcription profile of P/1C1 (genogroup 1) isolated from a patient diseased with a non A-G hepatitis was analysed. Four mRNAs were identified, which encoded the seven proteins ORF1, ORF1/1, ORF1/2, ORF2, ORF2/2, ORF3 and ORF4. Expression of the ORF1 protein and its splice variant ORF1/1 in cell culture was detected by an ORF1-specific antiserum. Analysis of N-terminal tagged P/1C1-encoded proteins revealed that ORF1, ORF1/1 and ORF1/2 were localised in the nucleoli, ORF3 and ORF4 resided in the nucleoplasm, ORF2/2 appeared either in the nucleoli or the whole nucleus while ORF2 was the only protein seen in the cytoplasm.

Percutaneous continuous aortic flow augmentation for cardiac recovery in a chronic heart failure patient with peripheral vascular disease.

Peripheral vascular disease is an obstacle to the use of continuous aortic flow augmentation (CAFA). The authors used CAFA in a patient with a 50% stenosis of the left iliac artery. Five hours after initiating therapy, flow rates dropped from 1.47 L/min to 0.2 L/min, possibly due to obstruction around the inflow cannula near the site of the iliac artery stenosis. Flow was stabilized by adequate fluid infusion and successfully restored by slightly withdrawing the tip of the inflow catheter. This finding suggests that peripheral vascular disease is a relative-not an absolute-contraindication for CAFA, but requires close monitoring of flow during CAFA therapy.

Molecular biology of Porcine circovirus: analyses of gene expression and viral replication.

The rep gene of Porcine circovirus type 1 directs the synthesis of two proteins. The full-length protein Rep is 312 amino acids in size, the spliced variant Rep' is truncated (168 aa) and exon 2 is frame-shifted. Replication of PCV1 DNA depends on synthesis of both proteins. Rep and Rep' bind in vitro to double-stranded DNA fragments comprising part of the origin of replication of PCV1, but the minimal binding sites of the two proteins are distinct. Rep protein represses the promoter of the rep gene by binding to the two inner hexamers H1 and H2. Although Rep' binds to the same sequence, it does not influence Prep. Twelve hours after PCV1 infection, similar amounts of rep and rep' were detected by real-time PCR, but later on, the ratio of the two transcripts varied. Both proteins are co-localised in the nucleus and formation of homo- and heteromeric complexes has been observed. When a replication assay was performed, in which Rep and Rep' protein of PCV1 was used to replicate the origin of PCV1 and PCV2, the rep gene products were found to initiate replication at both origins of replication.

New reporter gene-based replication assay reveals exchangeability of replication factors of porcine circovirus types 1 and 2.

Two types of porcine circovirus (PCV), which differ in their pathogenicity, are known. PCV type 2 (PCV2) is the etiological agent of postweaning multisystemic wasting syndrome in swine, while PCV1 has not yet been linked to a disease. Corroborating earlier observations in PCV1, transcript mapping revealed that the rep gene of PCV2 encodes two products, the full-length protein Rep and the spliced version Rep' and that the simultaneous expression of Rep and Rep' proteins is essential for initiation of replication of PCV2. The interchangeability of the replication factors of PCV1 and PCV2 was examined. The rep gene products of PCV2 were not only able to bind the PCV2 origin but also the origin of PCV1 and vice versa. To investigate the competence of the Rep/Rep' proteins to initiate replication at the heterologous origin, a new replication assay was developed. It measures the expression of a luc reporter gene present on a plasmid carrying the origin of the investigated replicon. Replication is initiated by expression of the appendant replicase from a second plasmid and results in replication of the origin plasmid coupled with an increase in the Luc activity. Using this method to compare replication of PCV1 and PCV2 in cell culture, it was shown that the Rep/Rep' protein of PCV2 initiated replication at the origin of PCV1, as did the reciprocal combination. Our results indicate that the cis- and trans-acting replication factors of the two viruses are functionally exchangeable.

Epiphyll deterrence to the leafcutter ant Atta cephalotes.

Epiphyll growth on leaves of the grapefruit Citrus paradisi and the understory cyclanth Cyclanthus bipartitus repelled the fungus-growing, leafcutter ant Atta cephalotes from harvesting leaves of these tropical plants. Experimental removal of epiphylls from leaves resulted in 2-3 times more herbivore damage by leafcutter ants as compared to matched leaves with epiphylls. Because of the protection from herbivore damage, host plants may derive a partial fitness benefit from association with epiphylls.