Site Of Penetration Research Articles

Margin details matter: The prognostic significance of pseudocapsule invasion at the site of involved margin in prostatectomy specimens

BackgroundAn involved surgical margin at prostatectomy has long been associated with elevated risk of prostate cancer recurrence; however, not all patients with an involved margin will relapse, and thus details of the involved margin may provide an opportunity for risk subset stratification. The present investigation seeks to determine whether a difference exists in recurrence rates when the margin involvement is at a site of prostate pseudocapsule invasion vs. within the prostate parenchyma proper. MethodsPatients were retrospectively identified for inclusion by clinically localized disease and prostate-specific antigen (PSA) level of<30ng/ml at diagnosis, managed with prostatectomy alone and identified to have involvement of surgical margin(s). Exclusion criteria were: pT3b or pN1 disease, immediate/nonsalvage postoperative radiation or hormone therapy, or insufficient follow-up (<12mo). Pathology slides were reviewed by a pathologist blinded to outcome, for determination of pseudocapsule invasion at a site of margin involvement. Disease recurrence was defined as PSA level of≥0.2ng/ml and rising, per contemporary guidelines. Kaplan-Meier method was used for construction of disease control estimate confidence intervals; Cox Proportional Hazards Model was used to compare disease control across groups. ResultsBetween 2003 and 2010, 155 patients were identified for inclusion in the present study. The median age was 61 years, and all had clinical stage T1 and T2 disease (75% T1c). At diagnosis, the Gleason score was 6, 7, and 8–9 for 103 (66%), 42 (27%), and 10 (6%) patients, respectively, with median PSA level of 5.6ng/ml (85%≤10). For 149 patients with reviewable margin site data, 51 (34%) demonstrated involvement within or beyond the pseudocapsule. At a median follow-up of 68 months (range: 13–137), 62 patients had experienced PSA relapse. The estimated 5-year PSA relapse rates for patients with an involved margin at the site of pseudocapsule invasion vs. prostate parenchyma were 49% vs. 34%, respectively (P = 0.017; hazard ratio = 1.853). ConclusionsEarly PSA relapse rates are high for patients with involved surgical margin(s) without seminal vesicle or node involvement at prostatectomy; however, for patients who are followed without immediate adjuvant therapy, presence of tumor cells at the margin in a site of pseudocapsule invasion or penetration confers a higher risk of recurrence.

Comparison of cell death and accumulation of reactive oxygen species in wheat lines with or without Yr36 responding to Puccinia striiformis f. sp. tritici under low and high temperatures at seedling and adult-plant stages

Yr36 is an important gene conferring resistance to stripe rust of wheat caused by Puccinia striiformis f. sp. tritici (Pst). To determine if the Yr36 resistance is correlated to reactive oxygen species (ROS) burst and cell death, wheat near-isogenic lines with (UC1041 + Yr36) and without (UC1041) the gene were histologically characterized for response to Pst infection. Yr36 conferred stripe rust resistance at both seedling and adult-plant stages when the gene line was tested with Pst race CYR29 at a high-temperature (HT) cycle (12 °C at night and 33 °C during the day). At the HT cycle, the growth of secondary hyphae was obviously suppressed in both seedlings and adult plants of UC1041 + Yr36 compared with those of UC1041. The percentages of infection sites with necrotic host cells in UC1041 + Yr36 were significantly higher than UC1041 60 hours after inoculation (hai) at both seedling and adult-plant stages. Mesophyll cell death in the inoculated UC1041 + Yr36 leaves at the HT cycle was stronger than at a low-temperature (LT) cycle (12 °C at night and 18 °C during the day). At the HT cycle, the level of ROS burst started increasing in the inoculated leaves of UC1041 + Yr36 when Pst hyphae started differentiating and extending, and simultaneously, the number of penetration sites with hypersensitive cell death was also increasing. The results indicate that Yr36 product affects the ROS accumulation and cell death of the host in interaction of wheat with Pst.

Cases of cutaneous anthrax in eastern Turkey: The reports of three cases

Anthrax is an acute disease caused by the bacterium Bacillus anthracis. This bacteria can form dormant endospores. When spores are inhaled, ingested, or come into contact with a skin lesion on a host, they may become reactivated multiply and rapidly. B. anthracis bacterial spores are soil-borne. Because of their long lifespan, spores are present globally and remain at the burial sites of animals killed by anthrax for many decades. Diseased animals can spread anthrax to humans, either by direct contact or by consumption of a diseased animal’s flesh. The most frequent clinical type of anthrax is cutaneous anthrax. It presents as a boil-like skin lesion that eventually forms an ulcer with a black center (eschar). Cutaneous anthrax form often within the site of spore penetration between two and five days after exposure. Until the 20 th century, anthrax infections killed thousands of animals and people worldwide each year. Animal vaccination programs and antibiotic therapy were decreased the number of deaths. But, anthrax is still a problem in less developed countries. It has been reported that the incidence of disease has decreased in Turkey. However, we present here in three cases of cutaneous anthrax admitted to the emergency department of our hospital within a week.

Two kinds of ocular trauma score for paediatric traumatic cataract in penetrating eye injuries

PurposeTo compare the ocular trauma score (OTS) and the paediatric penetrating ocular trauma score (POTS) as prognostic model for visual outcome in paediatric traumatic cataract cases after penetrating eye injuries. MethodsAll children younger than 16 years with unilateral traumatic cataract following penetrating trauma between 2007 and 2012 were retrospectively reviewed. Univariate chi-square analysis was conducted to identify the variables associated with profound visual loss. The area under the receiver-operating characteristic curves (AUROC) was used to assess the predictive ability of the two models. ResultsThe study group comprised 65 boys and 37 girls. The variables associated with profound visual loss were: a relative afferent papillary defect (RAPD) (P<0.001), poor initial vision (P=0.01), vitreous haemorrhage (P<0.001), retinal detachment (P<0.001), posterior penetrating site (P<0.001), hyphema (P<0.001), no intraocular len implantation (P<0.001) and endophthalmitis (P=0.001). OTS could not be calculated in 21 patients (20.6%) without clinical data on initial visual acuity and RAPD. For the patients with complete clinical data, POTS was similar to OTS in predicting poor vision (AUROC 0.904 vs 0.924) and in predicting good vision (AUROC 0.766 vs 0.736). For all the samples, POTS was a robust predictor of poor vision (AUROC 0.910) and had a moderate ability to predict good vision (AUROC 0.764). ConclusionOTS has high ability to predict visual outcome for paediatric traumatic cataract following penetrating ocular trauma. POTS is also a reliable prognostic model for very young child without initial vision or RAPD, but is only for penetrating eye injuries.

ENHANCED DISEASE RESISTANCE4 associates with CLATHRIN HEAVY CHAIN2 and modulates plant immunity by regulating relocation of EDR1 in Arabidopsis.

Obligate biotrophs, such as the powdery mildew pathogens, deliver effectors to the host cell and obtain nutrients from the infection site. The interface between the plant host and the biotrophic pathogen thus represents a major battleground for plant-pathogen interactions. Increasing evidence shows that cellular trafficking plays an important role in plant immunity. Here, we report that Arabidopsis thaliana ENHANCED DISEASE RESISTANCE4 (EDR4) plays a negative role in resistance to powdery mildew and that the enhanced disease resistance in edr4 mutants requires salicylic acid signaling. EDR4 mainly localizes to the plasma membrane and endosomal compartments. Genetic analyses show that EDR4 and EDR1 function in the same genetic pathway. EDR1 and EDR4 accumulate at the penetration site of powdery mildew infection, and EDR4 physically interacts with EDR1, recruiting EDR1 to the fungal penetration site. In addition, EDR4 interacts with CLATHRIN HEAVY CHAIN2 (CHC2), and edr4 mutants show reduced endocytosis rates. Taken together, our data indicate that EDR4 associates with CHC2 and modulates plant immunity by regulating the relocation of EDR1 in Arabidopsis.

The β‐amylase genes: negative regulators of disease resistance for Verticillium dahliae

Little is known about the role of plant primary metabolism in defence against pathogens. The present study is the first investigation published that examines the role of β‐amylase (BAM) genes upon fungal, Verticillium dahliae, infection. The responses of Arabidopsis thaliana plants impaired in BAM1, BAM2, BAM3, BAM4 genes, along with double, triple and quadruple mutants of those genes, were used to explore the involvement of BAM in the host plant–V. dahliae interaction. Less severe symptoms were recorded in bam mutants compared to wild type. Real‐time quantitative PCR (qPCR) revealed that the decrease in symptom severity shown in bam plants was correlated with reductions in the growth of the pathogen in the plants. Confocal microscopy of the most and least susceptible bam mutants and the wildtype plants showed that there were no differences between them in the number of attached conidia and penetration sites on the roots. BAM1, BAM2 and BAM3 expression was altered upon V. dahliae infection in the aerial tissues of the wild type. Analysis by qPCR of the PR1 and PDF1.2 expression in the bam3, bam1234, bam14 and wildtype plants showed that PR1 was up‐regulated in the roots of bam plants upon V. dahliae infection.

Do margin details matter? The prognostic significance of capsule invasion at the site of involved margin in prostatectomy specimens.

131 Background: An involved surgical margin at prostatectomy has long been associated with elevated risk of prostate cancer recurrence; however, not all patients with an involved margin will relapse, and thus details of the involved margin may provide an opportunity for risk subset stratification. The present investigation seeks to determine whether a difference exists in recurrence rates when the margin involvement is at a site of prostate capsule invasion versus within the prostate parenchyma proper. Methods: Patients were retrospectively identified for inclusion by clinically localized disease and PSA &lt;30 at diagnosis, managed with prostatectomy alone and identified to have involvement of surgical margin(s). Exclusion criteria were: pT3b or pN1 disease, immediate/non-salvage post-operative radiation or hormone therapy, or insufficient follow-up (&lt;12 months). Pathology slides were reviewed by a pathologist blinded to outcome, for determination of capsule invasion at a site of margin involvement. Disease recurrence was defined as PSA &gt;0.2 and rising, per contemporary guidelines. Kaplan-Meier method was employed for disease control estimates. Results: Between 2003 and 2010, 155 patients were identified for inclusion in the present study. The median age was 61 years, and all had cT1-2 disease (75% T1c). At diagnosis, the Gleason score was 6, 7, and 8-9 for 103 (66%), 42 (27%), and 10 (6%) patients, respectively, with median PSA 5.6 (85% &lt;10). For 149 patients with reviewable margin site data, 51 (34%) demonstrated involvement within or beyond the capsule. At a median follow-up of 68 months (range 13-137), 62 patients had experienced PSA relapse. The estimated 5-year PSA relapse rates for patients with an involved margin at the site of capsule invasion versus prostate parenchyma were 49% versus 34%, respectively (p=0.015). Conclusions: Early PSA relapse rates for all patients with involved surgical margin(s) but negative seminal vesicles and nodes at prostatectomy are high; however, for patients who are followed without immediate adjuvant therapy, presence of tumor cells at the margin in a site of capsule invasion or penetration confers a higher risk of recurrence.

A nucleotide exchange factor promotes endoplasmic reticulum-to-cytosol membrane penetration of the nonenveloped virus simian virus 40.

The nonenveloped simian polyomavirus (PyV) simian virus 40 (SV40) hijacks the endoplasmic reticulum (ER) quality control machinery to penetrate the ER membrane and reach the cytosol, a critical infection step. During entry, SV40 traffics to the ER, where host-induced conformational changes render the virus hydrophobic. The hydrophobic virus binds and integrates into the ER lipid bilayer to initiate membrane penetration. However, prior to membrane transport, the hydrophobic SV40 recruits the ER-resident Hsp70 BiP, which holds the virus in a transport-competent state until it is ready to cross the ER membrane. Here we probed how BiP disengages from SV40 to enable the virus to penetrate the ER membrane. We found that nucleotide exchange factor (NEF) Grp170 induces nucleotide exchange of BiP and releases SV40 from BiP. Importantly, this reaction promotes SV40 ER-to-cytosol transport and infection. The human BK PyV also relies on Grp170 for successful infection. Interestingly, SV40 mobilizes a pool of Grp170 into discrete puncta in the ER called foci. These foci, postulated to represent the ER membrane penetration site, harbor ER components, including BiP, known to facilitate viral ER-to-cytosol transport. Our results thus identify a nucleotide exchange activity essential for catalyzing the most proximal event before ER membrane penetration of PyVs. PyVs are known to cause debilitating human diseases. During entry, this virus family, including monkey SV40 and human BK PyV, hijacks ER protein quality control machinery to breach the ER membrane and access the cytosol, a decisive infection step. In this study, we pinpointed an ER-resident factor that executes a crucial role in promoting ER-to-cytosol membrane penetration of PyVs. Identifying a host factor that facilitates entry of the PyV family thus provides additional therapeutic targets to combat PyV-induced diseases.

The endoplasmic reticulum membrane J protein C18 executes a distinct role in promoting simian virus 40 membrane penetration.

The nonenveloped simian virus 40 (SV40) hijacks the three endoplasmic reticulum (ER) membrane-bound J proteins B12, B14, and C18 to escape from the ER into the cytosol en route to successful infection. How C18 controls SV40 ER-to-cytosol membrane penetration is the least understood of these processes. We previously found that SV40 triggers B12 and B14 to reorganize into discrete puncta in the ER membrane called foci, structures postulated to represent the cytosol entry site (C. P. Walczak, M. S. Ravindran, T. Inoue, and B. Tsai, PLoS Pathog 10: e1004007, 2014). We now find that SV40 also recruits C18 to the virus-induced B12/B14 foci. Importantly, the C18 foci harbor membrane penetration-competent SV40, further implicating this structure as the membrane penetration site. Consistent with this, a mutant SV40 that cannot penetrate the ER membrane and promote infection fails to induce C18 foci. C18 also regulates the recruitment of B12/B14 into the foci. In contrast to B14, C18's cytosolic Hsc70-binding J domain, but not the lumenal domain, is essential for its targeting to the foci; this J domain likewise is necessary to support SV40 infection. Knockdown-rescue experiments reveal that C18 executes a role that is not redundant with those of B12/B14 during SV40 infection. Collectively, our data illuminate C18's contribution to SV40 ER membrane penetration, strengthening the idea that SV40-triggered foci are critical for cytosol entry. Polyomaviruses (PyVs) cause devastating human diseases, particularly in immunocompromised patients. As this virus family continues to be a significant human pathogen, clarifying the molecular basis of their cellular entry pathway remains a high priority. To infect cells, PyV traffics from the cell surface to the ER, where it penetrates the ER membrane to reach the cytosol. In the cytosol, the virus moves to the nucleus to cause infection. ER-to-cytosol membrane penetration is a critical yet mysterious infection step. In this study, we clarify the role of an ER membrane protein called C18 in mobilizing the simian PyV SV40, a PyV archetype, from the ER into the cytosol. Our findings also support the hypothesis that SV40 induces the formation of punctate structures in the ER membrane, called foci, that serve as the portal for cytosol entry of the virus.

Combination of heat treatment and chitosan coating to improve postharvest quality of wolfberry (Lycium barbarum)

SummaryThe synergistic application of hot water dip at 42 °C for 30 min and 1% chitosan coating on differentiation in postharvest quality traits, microstructure as well as microbiological evolution of wolfberry fruits was investigated. Fresh wolfberry fruits were stored at 2 ± 0.5 °C and 90% relative humidity (RH) for 28 days. Results indicated the combination of prestorage heat treatment and chitosan coating maintained higher levels of ascorbic acid, total phenolic contents and antioxidant capacity as well as lower decay, compared with untreated wolfberry fruits. The possible mechanism was that the heat treatment almost sealed open stomata to limit the sites of pathogen penetration into fruits independently, followed by the biofilm formed by chitosan which controlled secondary infections as well as slowed changes in fruit respiration and metabolic activity in wolfberries. The synergistically treated fruit also exhibited a higher acceptability obtained by sensory analysis after cold storage. In this sense, the integrated application of heat treatment and chitosan coating could be regarded as an effective strategy to extend storage life and maintain the postharvest quality of wolfberry fruits.

Third-stage nematode larvae of Contracaecum osculatum from Baltic cod (Gadus morhua) elicit eosinophilic granulomatous reactions when penetrating the stomach mucosa of pigs.

Third-stage larvae of the anisakid nematode Contracaecum osculatum were recovered from livers of Atlantic cod (Gadus morhua) caught in the Baltic Sea (June 2014) and used for experimental infection of two pigs (one male and one female). Each pig received 215 larvae by oral infection (feeding with minced cod liver containing live nematode larvae). Pigs were euthanized after 5 days, necropsied, and subjected to parasitological investigation. A total of 12 larvae were found penetrating the mucosa of the ventricle (7 in the female pig and 5 in the male pig) eliciting a granulomatous reaction at the penetration site. Four non-attached larvae were found in the female pig stomach and one in the male pig. Petechial bleeding was observed at several locations in the ventricular mucosa where larvae were located. Histological examination of the stomach mucosa revealed a massive cellular infiltration (giant cells, lymphocytes, macrophages, granulocytes, and fibroblast like cells) around the penetrating larva. Mononuclear and polymorphonuclear cells containing eosinophilic granulae were particularly prominent in the granulomas. Reactions correspond to reactions in pigs following experimental infection with the human pathogenic anisakid larvae Anisakis sp. and Pseudoterranova sp. which suggests that C. osculatum might have a zoonotic potential as well.

Evidence for Systemic Infection by Puccinia horiana, Causal Agent of Chrysanthemum White Rust, in Chrysanthemum.

Puccinia horiana, causal agent of the disease commonly known as chrysanthemum white rust (CWR), is a quarantine-significant fungal pathogen of chrysanthemum in the United States and indigenous to Asia. The pathogen was believed to have been eradicated in the United States but recently reappeared on several occasions in northeastern United States. The objective of the study presented here was to determine whether P. horiana could systemically infect chrysanthemum plants, thus providing a means of survival through winters. Scanning and transmission electron microscopy revealed the development of P. horiana on the surface and within leaves, stems, or crowns of inoculated chrysanthemum plants artificially exposed to northeastern U.S. winter temperatures. P. horiana penetrated leaves directly through the cuticle and then colonized the mesophyll tissue both inter- and intracellularly. An electron-dense material formed at the interface between fungal and host mesophyll cells, suggesting that the pathogen adhered to the plant cells. P. horiana appeared to penetrate mesophyll cell walls by enzymatic digestion, as indicated by the absence of deformation lines in host cell walls at penetration sites. The fungus was common in vascular tissue within the infected crown, often nearly replacing the entire contents of tracheid cell walls. P. horiana frequently passed from one tracheid cell to an adjacent tracheid cell by penetration either through pit pairs or nonpitted areas of the cell walls. Individual, presumed, fungal cells in mature tracheid cells of the crown and stems arising from infected crowns suggested that the pathogen might have been moving at least partially by means of the transpiration stream. The demonstration that chrysanthemum plants can be systemically infected by P. horiana suggests that additional disease control measures are required to effectively control CWR.

The structural sheath protein of aphids is required for phloem feeding

Aphids produce two types of saliva that mediate their interactions with plants. Watery saliva is secreted during cell penetration and ingestion, whereas gel saliva is secreted during stylet movement through the apoplast where it forms a sheath around the stylet to facilitate penetration and seal puncture sites on cell membranes. In order to study the function of the sheath when aphids interact with plants, we used RNA interference (RNAi) to silence the aphid structural sheath protein (SHP) in the pea aphid Acyrthosiphon pisum. The injection of 50 ng of double stranded RNA completely disrupted sheath formation, as confirmed by scanning electron microscopy. Aphid behavior was monitored using the electrical penetration graph technique, revealing that disrupted sheath formation prevented efficient long-term feeding from sieve tubes, with a silencing effect on reproduction but not survival. We propose that sealing the stylet penetration site in the sieve tube plasma membrane is part of a two-step mechanism to suppress sieve-tube occlusion by preventing calcium influx into the sieve tube lumen. The SHP is present in several aphid species and silencing has a similar impact to aphid-resistant plants, suggesting that SHP is an excellent target for RNAi-mediated pest control.

Attachment, penetration and early host defense mechanisms during the infection of filamentous brown algae by Eurychasma dicksonii.

Eurychasma dicksonii is one of the most common and widespread marine pathogens and attacks a broad spectrum of more than 45 brown algal species. The present study focuses on the mechanism used by the pathogen to attach on the host cell wall and force its way into algal cells. Ultrastructural examination revealed a needle-like structure which develops within the attached spore and extends along its main axis. Particular cell wall modifications are present at the basal part of the spore (adhesorium pad) and guide the needle-like tool to penetrate perpendicularly the host cell wall. The unique injection mechanism is shared with Haptoglossa species which suggests that this is an important characteristic of early diverging oomycetes. Furthermore, the encystment and adhesion mechanism of E. dicksonii shows significant similarities with other oomycetes, some of which are plant pathogens. Staining and immunolabelling techniques showed the deposition of β-1,3-glucans on the host cell wall at the pathogen penetration site, a strategy similar to physical responses previously described only in infected plant cells. It is assumed that the host defense in terms of callose-like deposition is an ancient response to infection.

Stabilized triple procedure for management of coexisting corneal opacity and cataract

We describe a surgical technique combining penetrating keratoplasty and cataract surgery with a secure anterior chamber to manage coexisting corneal opacity and cataract. Lamellar corneal dissection is performed to 80% of the corneal depth to provide sufficient visibility for cataract surgery. Manual small-incision cataract extraction and intraocular lens implantation is performed through a frown-shaped scleral tunnel. Subsequently, 4 small penetrating incisions are made along the edge of the trephination mark at the 3, 6, 9, and 12 o'clock positions. A graft is placed on the ophthalmic viscosurgical device-coated stromal bed, and the 4 penetration sites are sutured. The stromal bed is cut along the trephination groove, the graft is sutured quadrant by quadrant, and the stromal bed is drawn out of the anterior chamber. The surgery is completed with 16 sutures.

Anatomy of the diaphyseal nutrient foramen in the long bones of the pectoral limb of German Shepherds

This study investigated the anatomy of the nutrient foramen (NF) in German Shepherds by recording the number, site, position, and direction of penetration of the nutrient canal (NC) in the humerus, radius, and ulna of 50 individuals. The site index of the nutrient foramen (SI) was calculated as the ratio of the length to the NF site from the proximal end to the greatest length of the bone. The NF diameter was measured using different sized needles. Most humeri had only one NF on the caudal surface, particularly on the lateral supracondylar crest, or distal cranial surface. All radii had one NF, usually on the caudal surface, while most ulnae had one NF located on either the cranial or lateral surfaces. The SI and NF diameters were 58.0~59.5% and 0.73~0.78 mm in the humerus, 30.4~30.9% and 0.74~0.76 mm in the radius, and 29.3~29.8% and 0.67~0.68 mm in the ulna, respectively. With the exception of the relatively proximal NF of the radius, the direction of penetration followed Berard's rule. This study provides novel information on the location and diameter of the NF and direction of the NC in the long bones of the pectoral limb of German Shepherds.

Vascular Supply of the Cerebral Cortex is Specialized for Cell Layers but Not Columns

The vascular supply to layers and columns was compared in macaque primary visual cortex (V1) by labeling red blood cells via their endogenous peroxidase activity. Alternate sections were processed for cytochrome oxidase to reveal "patches" or "blobs," which anchor the interdigitated column systems of striate cortex. More densely populated cell layers received the most profuse blood supply. In the superficial layers the blood supply was organized into microvascular lobules, consisting of a central venule surrounded by arterioles. Each vessel was identified as an arteriole or venule by matching it with the penetration site where it entered the cortex from a parent arteriole or venule in the pial circulation. Although microvascular lobules and cytochrome oxidase patches had a similar periodicity, they bore no mutual relationship. The size and density of penetrating arterioles and venules did not differ between patches and interpatches. The red blood cell labeling in patches and interpatches was equal. Moreover, patches and interpatches were supplied by an anastomotic pial arteriole system, with no segregation of blood supply to the two compartments. Often a focal constriction was present at the origin of pial arterial branches, indicating that local control of cortical perfusion may be accomplished by vascular sphincters.

Role of strawberry volatile organic compounds in the development of Botrytis cinerea infection

Botrytis cinerea, the main pathogen of strawberry, has the ability to remain quiescent in unripe tissue and develop disease symptoms in ripe fruit. As strawberry ripening is characterized by an increase of aroma compounds, the role of volatile emission in the development of infection was investigated. Thirty‐five strawberry volatile organic compounds (VOCs) were tested on B. cinerea in vitro and volatile emission was analysed in strawberry harvested at four ripening stages by headspace solid‐phase microextraction/gas chromatography–mass spectrometry and proton transfer reaction–time of flight–mass spectrometry. The coupling of such data sets made it possible to conclude that key strawberry aroma compounds stimulate B. cinerea conidial germination and some typical wound‐volatiles stimulate pathogen conidial germination or mycelial growth. This study is the first report of fungal stimulation by some VOCs naturally occurring in strawberry: the esters ethyl butanoate, cis‐3‐hexenyl acetate, trans‐2‐hexenyl acetate, methyl butanoate and hexyl butanoate, the furanones furaneol and mesifurane, and the alcohol trans‐2‐hexenol. The results of this work provide advances in understanding the functional role of fruit VOCs and suggest, for the first time, that fruit VOCs may influence the development of B. cinerea from the latent phase and that they could favour the invasive growth of B. cinerea after wounding. In particular, ethyl butanoate and furaneol could signal strawberry ripening, and the green leaf volatiles trans‐2‐hexenol, trans‐2‐hexenyl acetate and cis‐3‐hexenyl acetate could signal the presence of damaged tissues that are easier sites for penetration by B. cinerea.

Use of Pre-Injection Diffusion of Local Anaesthetic as a Means of Reducing Needle Penetration Discomfort.

To determine if pre-injection diffusion of local anaesthetic solution influences the discomfort of needle penetration in the palate. A placebo-controlled, randomised, double-blind split-mouth investigation was conducted. 25 healthy adult volunteers were recruited and each received two needle penetrations in a random order during one visit. The penetration sites were 1 cm from the gingival margin of the first maxillary premolars on each side of the mouth. 30 gauge-13 mm needles which were attached to syringes that contained either 2% lidocaine with 0.125mg/ml epinephrine or physiological saline were used. For each penetration an operator encouraged a drop of solution to appear at the end of the needle and placed this drop with the bevel of the needle flat on the palate for 20 seconds. The discomfort was noted on a 100 mm visual analogue scale with end points marked "No pain" and "Unbearable pain". There was no significant difference in penetration discomfort between solutions, (mean VAS = 26.80±19.36mm for lidocaine and 26.20±18.39mm for saline) however the 2nd penetration was significantly more uncomfortable than the first (mean VAS = 31.00±19.84 mm and 22.00±16.65 mm respectively). Pre-injection diffusion of local anaesthetic solution did not influence the discomfort of needle penetration in the palate.

Development of a New Device for Synovial Biopsies.

Synovial biopsy is a reliable procedure that permits the identification of specific pathologies. Currently available needles for blind synovial biopsy usually consist of 2 components. We designed a novel 1-piece device for blind and minimal invasive synovial biopsy of the knee joint. A convex-shaped trocar with internal sharp plunger at the distal end and a fluid channel was engineered. Synovial biopsy of the suprapatellar recessus of the knee was performed in 8 different cadavers. The intra-articular position of the tip was confirmed by aspiration of prior injected saline fluid. The trocar was levered upward with the open notch of the device facing the anterior wall of the recessus. Then, the punch mechanism was closed and the device removed. Routine histology of the obtained tissue was performed. After the intervention, the joints were prepared for macroscopic inspection of the synovial tissue, including penetration and biopsy sites. Fifteen interventions were performed. In all cases, sufficient synovial tissue was obtained. Mean length of the biopsies was 2.4 mm (range 2.0-4.3 mm) and width was 2.0 mm (range 1.4-2.6 mm). Inside the suprapatellar recessus, the mean distance from the entry site of the device to the biopsy site was 3.8 cm (range 1.1-3.4 cm). Histological analysis confirmed synovial and capsule tissue in all cases. This new device is a potentially useful tool for quick synovial biopsy of the knee in the clinical setting.