Winter activity of Ixodes ricinus in a Berlin forest

Nymphs and adults of the hard tick Ixodes ricinus typically quest in Central Europe from March/April to October/early November. The recent trend towards milder winters raises the question whether I. ricinus will gradually extend its seasonal activity and use mild winter periods for questing. The present study followed the host-seeking activity of I. ricinus from early September 2006 to early March 2007 in a Berlin forest. Field-collected larval and nymphal ticks were allowed to take a full blood meal in the laboratory and to moult. The resultant unfed nymphs (n=125) and adults (n=149) were released onto three field plots (1×1 m²) in early September 2006. A second group of field-collected unfed I. ricinus nymphs (n=79) and adults (n=47) were released onto a field plot in early September 2006. Each plot was covered by an approximately 10 cm thick layer of leaf litter (mostly of oak and beech) to provide a humid microclimate refuge and shelter for non-active ticks. Wooden rods (60 cm high) arranged in a 6×8 grid on these field plots were climbed by questing nymphs and adults. Active ticks were observed on each of 33 observation dates. The winter activity revealed by this experiment was confirmed by flagging for ticks on two occasions in January and February 2007 with remarkably high numbers of collected ticks. This appears to be the first time that extended or even continuous winter activity of I. ricinus nymphs and adults has been demonstrated in Central Europe, most probably caused by the extremely mild winter of 2006/2007. The fact that I. ricinus now can be active during the whole winter, a time of the year when these ticks historically have been dormant when the weather is normal, is relevant to the public because people who enter forest areas should be well aware of the risk of getting tick bites and tick-borne infections in mild winter periods.     

The vertical distribution,density and the development of the tick Ixodes ricinus in mountain areas influenced by climate changes (The Krkonoše Mts., Czech Republic)

The altitudinal distribution and density of host-seeking Ixodes ricinus ticks were investigated by flagging in 2002–2006 along two vertical transects in the Krkonoše Mts. Comparison with historical data revealed that the I. ricinus upper distribution limit shifted from the former 700–800 m limit at least up to 1100 m above sea level (a.s.l.) during the last two decades. The tick thus has newly colonised the whole montane belt, covered predominantly with Norway spruce forests. The density of host-seeking nymphs decreased with increasing altitude: In the established mixed forests located at 600–800 m a.s.l., it reached the mean value of 31 nymphs per hour of flagging (with a maximum density of 125 nymphs per hour). The mean density attained 7.1 nymphs per hour (maximum 66) in 870–1020 m and 2 nymphs per hour (maximum 6.3) in the newly colonised areas at 1080–1100 m a.s.l. Regular records of host-seeking I. ricinus larvae and nymphs up to 1020 m a.s.l., together with rather regular records of nymphs and presence of single host-seeking adults up to 1100 m a.s.l. indicated viability of the newly established local tick populations. The present empirical upper limit was experimentally confirmed by the investigation of the developmental capability of eggs, fed larvae and nymphs along a vertical transect (650–1550 m a.s.l.) in 2004–2006. The experimental results demonstrated a strong relationship between tick developmental success and duration on the one hand and altitude on the other. They showed that climatic conditions up to 1100 m a.s.l. are at present suitable for the development of all tick stages. However, it also revealed that ticks were able to complete their development up to at least 1250 m a.s.l. in the unusually warm summer and autumn 2006 (the mean temperature for June–October 2006 exceeded the 1960–1990 mean value by 1.8 °C). The causes of tick expansion could be found when analysing long-term (1961–2005) climatic data from the Krkonoše Mts., which showed a systematic and significant rise of the mean annual air temperature by 1.4 °C at 1000 m a.s.l. The most pronounced temperature increase (mean monthly temperature by 1.5–3.5 °C at 1000 m a.s.l.) was determined in May–August, which is crucial for host finding and development of I. ricinus. Abundance of the newly settled local tick populations at higher altitudes may increase in the future after their closer establishment in local mountain ecosystems.     

Preventive effect of permethrin-impregnated clothing to Ixodes ricinus ticks and associated Borrelia burgdorferi s.l. in Germany

The protective effectiveness of a factory-based, permethrin-impregnated military battle dress uniform (BDU) trouser using a new high-residual, polymer-coating technique has been evaluated by field testing against Ixodes ricinus, a vector of Borrelia burgdorferi s.l. During 36 h exposure of test subjects walking in known tick-infested habitats in the Kühkopf mountain area, Koblenz, Germany, between June and October 2006, 6 I. ricinus were found crawling with a visible excitatory effect on those legs covered with fabric impregnated with 1200 mg permethrin/m², whereas 132 ticks were collected from the negative control legs. B. burgdorferi s.l. infection was detected in 33% (2/6) adult male, 56% (9/16) adult female, 11% (6/56) nymphal, and 0% (0/46) larval I. ricinus sampled from the negative control legs. The few ticks collected from the impregnated fabric tested all negative for B. burgdorferi s.l. The mean tick infestation rate on the negative control legs of test subjects was 3.6±2.7 (mean±SD; range 0–12) per hour exposure. Permethrin-impregnated clothing conveyed a high mean protection rate of 95.5% against questing I. ricinus ticks, making it an excellent tool for the prevention of associated tick-borne diseases.     

Tick-borne encephalitis virus expansion to higher altitudes correlated with climate warming

Since 2002, the expansion of Ixodes ricinus ticks and tick-borne infection agents have been studied in the Krkonoše Mts., Czech Republic. Tick-borne encephalitis virus was detected by means of RT-PCR. In 2003, it was detected in 2 out of 491 ticks at 620 and 710–720 m a.s.l., respectively, and in 3 out of 939 ticks at 600 m a.s.l. at the same locality in 2004. In 2005, tick-borne encephalitis virus was detected in 5 out of 295 ticks at 900–1100 m a.s.l., which is above the formerly known altitudinal limit of I. ricinus distribution. The reason for that could be found in the changing climate. Based on the meteorological data collected in the Krkonoše Mts., 1961–2005, there was a significant increase in the mean annual temperature (1.3–1.4 °C) over that period, namely by 2–3.5 °C in May through August. Thus, with respect to the average vertical temperature gradient in summer of about 0.6 °C/100 m, 2 °C correspond to 300–350 m in altitude, and accordingly 3.5 °C correspond to a shift in altitude of approximately 550–600 m, that being in accordance with environmental conditions of the former I. ricinus altitudinal limit confirmed in the Krkonoše Mts. 20 years ago.     

Sudden increase in tick-borne encephalitis cases in the Czech Republic, 2006

In the year 1993, there was a great increase in the number of tick-borne encephalitis (TBE) cases in the Czech Republic, and this high level has persisted with certain fluctuations. In the period 1965–1992, there were 8690 cases (annual average: 310.4), whereas there were 8674 registered cases in the years 1993–2006 (annual average: 619.6). In 2006, there was an exceptionally sudden increase with 1029 registered TBE cases, i.e., the national incidence was 10/100,000, the highest level recorded so far. It is documented that this situation was significantly influenced by exceptional weather in 2006. During the warm and rainy spring and early summer period (April–June), high host-seeking activity of Ixodes ricinus was recorded and also a high incidence of TBE resulting in a typical spring/summer peak of disease (June/July). There was a sudden break in the weather and a cool August followed (average temperature 0.8 °C below the long-term [30 years] average) with high precipitation (175% above the long-term average). This had a favourable effect on the activity of I. ricinus, which did not decline in August (as it has been typical in other years), and it only gradually decreased thereafter in the warm autumn (September, October, and November average monthly temperatures 3.2, 2.8, and 3.2 °C above the long-term average, respectively). Also the recreational behaviour of people was influenced very much in the peak period of summer vacations and school vacations (TBE typically is a recreational infection in the Czech Republic). August weather resulted in a decline in water sports and directed the majority of holidaymakers to go on a mushroom foray, a traditional national hobby in the Czech Republic. This was also triggered by that year's very successful mushroom season, which was repeatedly highlighted by the media. This resulted in a second (late summer/autumn) peak of the incidence curve of TBE, which even exceeded the spring/summer culmination. Such a type of TBE seasonality had not been observed in the Czech Republic before, resulting in an exceptionally high overall annual incidence.     

Distribution of Rickettsia helvetica in Ixodes ricinus tick populations in Poland

Questing Ixodes ricinus ticks from different regions of Poland were investigated for the presence of spotted fever group (SFG) rickettsiae. A total of 1214 DNA samples of 2813 ticks, including 820 individual adult ticks and 394 pools containing 1993 nymphs, were tested by PCR for a fragment of the rickettsial gltA gene using the primers RpCs.877 and RpCs.1258. Overall, at least 5.5% ticks were found to be positive with the highest prevalence observed in females (10.6%). A sample of 14 positive PCR products was sequenced. Analyzed fragments of 270–370 bp showed 100% similarities to corresponding sequences of Rickettsia helvetica deposited in the GenBank. Results of our investigation confirm the occurrence and wide distribution of R. helvetica in I. ricinus tick populations in Poland. This rickettsia should be added to the list of potentially dangerous pathogens transmitted by ticks in our country.     

Paediatric tick-borne encephalitis (TBE) vaccines: Schedules to optimise protection

Although tick-borne encephalitis (TBE) vaccines have been used successfully for decades, direct comparisons of different immunisation schedules and TBE vaccines are limited. This randomised controlled study compared two paediatric TBE vaccines (Encepur^® Children and FSME-IMMUN^® 0.25 ml Junior), administered according to a conventional (days 0, 28, and 300) or an accelerated (days 0, 14, and 300) schedule; all subjects received Encepur^® Children as the third vaccine dose on day 300. With both vaccines, superior immunogenicity was achieved using the conventional schedule, compared with the accelerated schedule, as assessed at days 42, 300, and 321, by geometric mean titres calculated using the neutralisation test (NT). Overall, the immune response to vaccination, assessed by NT, was greater in children vaccinated with Encepur^® Children than in those receiving FSME-IMMUN^® 0.25 ml Junior, for both the conventional and accelerated schedule. These differences were statistically significant for all post-vaccination evaluation days (42, 300, and 321; p<0.001). In conclusion, the conventional schedule is superior to the accelerated schedule. Furthermore, vaccination with Encepur^® Children offers superior immunogenicity over FSME-IMMUN^® 0.25 ml Junior, using either the conventional or accelerated schedule.     

Toward verified and validated FE simulations of a femur with a cemented hip prosthesis

BackgroundVerified and validated CT-based high-order finite element (FE) methods were developed that predict accurately the mechanical response of patient-specific intact femurs. Here we extend these capabilities to human femurs undergoing a total hip replacement using cemented prostheses.MethodsA fresh-frozen human femur was CT-scanned and thereafter in vitro loaded in a stance position until fracture at the neck. The head and neck were removed and the femur was implanted with a cemented prosthesis. The fixed femur was CT-scanned and loaded through the prosthesis so that strains and displacements were measured. High-order FE models based on the CT scans, mimicking the experiments, were constructed to check the simulations prediction capabilities.ResultsThe FE models were verified and results were compared to the experimental observations. The correlation between the experimental and FE strains and displacements were (R² = 0.97, EXP = 0.96FE + 0.02) for the intact femur and (R² = 0.90, EXP = 0.946FE + 0.0012) for the implanted femur. This is considered a good agreement considering the uncertainties encountered by the heavy distortion embedded in the CT scan of the metallic prosthesis.DiscussionThe patient-specific FE model of the fresh-frozen femur with the cemented metallic prosthesis showed a good correlation to experimental observations, both when considering surface strains, displacements and strains on the prosthesis. The relatively short timescale to generate and analyze such femurs (about 6 h) make these analyses a very attractive tool to be used in clinical practice for optimization prostheses (dimensions, location and configuration), and allow to quantify the stress shielding.     

Characterization of tetracycline and methicillin resistant Staphylococcus aureus strains in a Spanish hospital: Is livestock-contact a risk factor in infections caused by MRSA CC398?

Tetracycline-resistance (Tet^R) has been postulated as a marker of the livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) lineage CC398. Objectives of the study: to determine the spa-types and assigned MLST clonal complexes (CCs) among all 98 MRSA-Tet^R strains recovered during 2011–2012 (from different patients) in a Spanish Hospital, analyzing the possible correlation with livestock-contact of the patients. All 98 strains were assigned to 9 CCs: CC398 (60.2%), CC1 (19.4%), CC5 (12.2%), and other CCs (8.2%). The 98 patients were classified into three groups: (A) contact with livestock-animals (n = 25); (B) no-contact with livestock-animals (n = 42); (C) no information about animal contact (n = 31). A significant higher percentage of CC398 strains was obtained in group A (76%) than in group B (50%) (p < 0.05), being the percentage in group C of 61.3%. Most of MRSA-Tet^R-CC398 strains presented a multi-resistance phenotype, including erythromycin, clindamycin, and ciprofloxacin, and the most prevalent detected genes were tet(M) and erm(C). Three strains presented the phenotype macrolide-susceptibility/lincosamide-resistance and contained the vga(A) gene. MRSA-CC1 strains showed higher percentages of erythromycin/clindamycin resistance (95%/89%) than MRSA-CC398 strains (58%/63%), and this resistance was usually mediated by erm(C) gene. Most of MRSA-CC5 strains showed resistance to ciprofloxacin, tobramycin/kanamycin and erythromycin. None of the strains presented the genes lukF/lukS-PV, tsst-1, eta, etb or etd. All MRSA-CC398 strains lacked the genes of the immune-evasion-cluster, but MRSA-CC1 strains carried these genes (type E). In conclusion, although MRSA CC398 is detected in a significant higher proportion in patients with livestock-contact; its detection in people without this type of contact also indicates its capacity for human-to-human transmission.     

An in vitro study on the maturation of conventional glass ionomer cements and their interface to dentin

The objective of the study was to investigate the influence of long-term storage (up to 1 year) and coating on the variation of micro-mechanical properties of four conventional restorative glass ionomer cements (GICs) within 3.5 mm deep class I cavities. Four commercially available GICs (Riva Self Cure (SDI), ChemFil Rock (Dentsply), Fuji IX Fast and Fuji IX GP Extra/Equia (GC)) were applied to 100 teeth. In each tooth, two similar 3.5 mm deep class I cavities were prepared and filled with the GICs, with and without resin coating. The samples were stored in artificial saliva at 37 °C for 1 week, 1 month, 3 months, 6 months and 1 year. The variation in mechanical properties (indentation modulus (E) and Vickers hardness (HV)) were determined in 100 μm steps starting from the filling surface, through the intermediate layer in between dentine and GIC, and ending 100 μm in dentin. HV and E were strongly influenced by the material (P < 0.05, partial eta-squared η_P² = 0.31 and 0.23) but less by aging duration (P < 0.05, η_P² = 0.02 and 0.12) and resin coating (P < 0.05, η_P² = 0.02 and 0.03). The depth of measurement (0–2 mm) has no influence on HV (P = 0.789). HV shows a gentle increase over the 1 year storage period (P = 0.002). A ～300 μm GIC zone at the areas close to dentin with weaker properties as those measured in dentin or GIC was identified in all fillings, irrespective of the presence of coating, and at all storage periods. The thickness of this zone is more strongly influenced by storage (P < 0.05, η_P² = 0.081) than by material type (P < 0.05, η_P² = 0.056), while coating showed no influence (P = 0.869). Filler morphology and dimension were similar to upper parts of the GIC filling; however, the amount of low cations was higher. We concluded that the development of an intermediate layer in between dentine and GIC with lower mechanical properties might be responsible for the bond quality of GIC to dentine. Moreover, class I GIC restorations are unlikely to feature constant mechanical properties throughout the cavity, regardless of conditions such as aging and coating.     

Essential oils and their components in combating fungal pathogens of animal and human skin

ObjectiveThe most common problems of modern medicine include fungal infections of the skin and its appendages caused by dermatomycetes, yeast-like fungi and moulds. Due to toxicity of pharmacological fungicides and promotion of ecology, natural substances with high antifungal properties are sought. Essential oils and their components show potential in this regard.Material and methodFourteen commercial essential oils were tested for antifungal activity. The study were carried out by agar dilution method against the following fungal species: Microsporum gypseum, Microsporum canis, Trichophyton mentagrophytes, Trichophyton violaceum, Aspergillus niger, Scopulariopsis brevicaulis and (IZ 1) dog skin isolate. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) values were determined for all tested oils and individual components of oils with the highest activity against the tested fungi. GC-MS analysis was performed for the most active oils.ResultsCinnamon, thyme, clove, geranium and manuka oils were most active against the test fungi. The MIC values in the tested oils were in the range of 0.5– > 10 μg/μl and the MFC amounted to 1.25– > 10 μg/μl. Activity of individual components against the tested fungi strains was selective and clearly higher against dermatomycetes fungi and IZ 1 isolate. The strongest effect on dermatomycetes fungi was of cinnamaldehyde, thymol and carvacrol–with the values of 0.039–1.25 μg/μl (MIC) and 0.078–1.25 μg/μl (MFC). In turn, the highest activity against IZ 1 isolate was of cinnamaldehyde, eugenol, carvacrol, geraniol and thymol. Their MIC and MFC was 0.313–1.25 μg/μl.ConclusionThe results prove the applicability of selected oils and their components as alternatives to synthetic agents in combating fungal dermatoses in animals and humans.     

Mechanisms of quinolone resistance in Aeromonas species isolated from humans,water and eels

Mechanisms of resistance were determined in 33 quinolone-resistant isolates of the species Aeromonas hydrophila, Aeromonas caviae, Aeromonas media, Aeromonas salmonicida, Aeromonas popoffii and Aeromonas veronii, recovered from humans, freshwater and eels. The quinolone resistance-determining regions (QRDRs) of gyrA and parC genes were sequenced in these resistant strains, as well as in 8 quinolone-sensitive Aeromonas used as controls. All quinolone-resistant Aeromonas carried point mutations in the gyrA QRDR at codon 83, respectively giving rise to substitutions Ser⁸³ → Ile (32 strains) or Ser⁸³ → Val (1 strain). Almost half of these isolates (48%) carried additional point mutations in the gyrA QRDR at codon 92 and/or in the parC QRDR at codon 80 corresponding to substitutions Leu⁹² → Met and Ser⁸⁰ → Ile. In all cases, MICs of quinolones were determined in the presence and absence of the efflux pump inhibitor phenylalanine-arginine β-naphthylamide (PAβN). Addition of PAβN had no effect on the levels of resistance observed in these isolates. In conclusion, the mechanism of quinolone resistance in the Aeromonas isolates studied was related to mutations in QRDR regions of gyrA and parC genes, with little obvious involvement of pumps inhibited by PAβN.     

Risk factors for blood transfusion in Cesarean section: A systematic review and meta-analysis

ObjectiveThe current study has been conducted to identify the risk factors associated with blood transfusion in women undergoing cesarean section (C-section). A detailed account of the risk factors associated withblood transfusion will ultimately prevent unnecessary crossmatching in hospitals , leading to the conservation of declining blood supplies and resources without subjugating the quality of care.Material and methodsWe performed a rigorous literature search using electronic databases, including PubMed, Cochrane CENTRAL, and Embase, for studies evaluating the risk factors for blood transfusion in C-section published until March 31, 2021. The Newcastle-Ottawa Quality Assessment Scale was deployed to assess the methodologic quality of the included studies. Mean differences (MD) and odds ratios (OR) with 95% confidence intervals were calculated using Review Manager version 5.3.ResultsThe search yielded 1563 records, 22 of which were eligible for inclusion, representing 426,094 women (10,959 in the transfused group and 415,135 in the non-transfused group). Participants in the transfused group had lower mean preoperative hematocrit (MD = ?3.71 [?4.46, ?2.96]; p < 0.00001; I² = 88%). Placenta previa (OR = 9.54 [7.23, 12.59]; p < 0.00001; I² = 88%), placental abruption (OR = 6.77 [5.25, 8.73]; p < 0.00001; I² = 72%), emergency C-section (OR = 1.92 [1.42, 2.60]; p < 0.0001; I² = 75%), general anesthesia (OR = 8.43 [7.90, 9.00]; p < 0.00001; I² = 72%), multiple gestations (OR = 1.60 [1.24, 2.06]; p = 0.0003; I² = 85%), preterm labor (OR = 3.34 [2.75, 4.06]; p < 0.00001; I² = 85%), prolonged labor (OR = 1.68 [1.44, 1.96]; p < 0.00001; I² = 78%), unbooked cases (OR = 2.42 [1.22, 4.80]; p = 0.01; I² = 80%), hypertensive disorders of pregnancy (OR = 1.81 [1.72, 1.90]; p < 0.00001; I² = 71%), and fibroids (OR = 2.32 [1.55, 3.47]; p < 0.0001; I² = 72%) were significantly higher in the transfused group compared to the non-transfused group. Chronic hypertension (OR = 0.67 [0.29, 1.55]; p = 0.36; I² = 90%), maternal age (MD = 0.09 [?0.27, 0.45]; p = 0.62; I² = 50%), maternal body mass index (MD = ?0.14 [?0.81, 0.53]; p = 0.67, I² = 86%), diabetes (OR = 0.93 [0.75, 1.15]; p = 0.51; I² = 52%), and malpresentation (OR = 0.65 [0.38, 1.11]; p = 0.13; I² = 64%) were not significantly associated with an increased risk of blood transfusion in C-section in the two groups.ConclusionPlacenta previa, placental abruption, emergency C-section, booking status, multiple gestations, and preoperative hematocrit were the risk factors most significantly associated with blood transfusion, while a prior C-section did not increase the risk of transfusion.     

Peri-implant reactivity and osteoinductive potential of immobilized rhBMP-2 on titanium carriers

Recombinant human BMP-2 (rhBMP-2) was immobilized non-covalently and covalently as a monolayer on plasma vapour deposited (PVD) porous commercially pure titanium surfaces in amounts of 5–8 μg cm^?2, providing a ca. 10-fold increase vs. previously reported values [37]. Dissociation of the immobilized [¹²⁵I]rhBMP-2 from the surface occurred in a two-phase exponential decay: a first rapid phase (ca. 15% of immobilized BMP-2) with a half-life of 1–2 days and a second slow sustained release phase (ca. 85% of immobilized BMP-2) with a half-life of 40–60 days. Dissociation rate constants of sustained release of k_?1 = 1.3–1.9 × 10^?7 s^?1 were determined, allowing an estimation of the binding constants (K_A) for the adsorbed rhBMP-2 monolayer, to be around 10¹² M^?1. The rhBMP-2-coated surfaces showed a high level of biological activity, as demonstrated by in vitro epifluorescence tests for alkaline phosphatase with MC3T3-E1 cells and in vivo experiments. In vivo osteoinductivity of rhBMP-2-coated implants was investigated in a gap-healing model in the trabecular bone of the distal femur condylus of sheep. Healing occurred without inflammation or capsule formation. The calculated concentration of released rhBMP-2 in the 1 mm gap ranged from 20 to 98 nM – well above the half-maximal response concentration (K_0.5) for inducing alkaline phosphatase in MC3T3-E1 cells. After 4, 9 and 12 weeks the bone density (BD) and bone-to-implant contact (BIC) of the explanted implants were assessed histomorphometrically. Implants with immobilized rhBMP-2 displayed a significant (2- to 4-fold) increase in BD and BIC values vs. negative controls after 4–9 weeks. Integration of implants by trabecular bone was achieved after 4 weeks, indicating a mean “gap-filling rate” of ～250 μm week^?1. Integration of implants by cortical bone was observed after 9 weeks. Control implants without rhBMP-2 were not osseointegrated. This study demonstrates the feasibility of enhancing peri-implant osseointegration and gap bridging by immobilized rhBMP-2 on implant surfaces which may serve as a model for future clinical applications.     

Dermatomycosis: comparison of the performance of calcofluor and potassium hydroxide 30% for the direct examination of skin scrapings and nails

Dermatomycosis is a common problem in clinical practice. A definitive diagnosis should be made before initiation of antifungal therapy. Mycological diagnosis is usually based on direct microscopy using potassium hydroxide and fungal culture. The aim of our study was to compare the performance of potassium hydroxide 30% (KOH) with slight heating and staining with 0.01% calcofluor (Fluorescent brightener 28^®) for the detection of fungi by direct microscopy. We evaluated prospectively 241 samples from patients suspected to have dermatomycosis (159 nail samples, 82 skin scrapings). Mycological culture was chosen as the reference method to assess the performance of each test. Out of the 241 samples, 152 mycological cultures were positive, of which 98 were classified as pathogenic fungi (86 dermatophytes, 11 non-dermatophyte molds, 1 Candida albicans). The sensitivity of direct microscopy was significantly higher with calcofluor than with KOH (respectively, 88% and 72%, P = 0.0116). Specificity was 93% with calcofluor and 88% with KOH (P = NS). Calcofluor showed higher specificity on skin scraping than on nails samples (respectively, 100% and 89%, P = 0.0138) while KOH specificity did not differ as a function of the type of specimen (respectively, 92% and 86%). In conclusion, calcofluor is a more sensitive and specific technique than KOH for the microscopic detection of dermatophytes and non-dermatophyte pathogenic fungi in nails and skin scrapings. The results are easy to read by inexperienced staff.     

Can a 15 m-overground wheelchair sprint be used to assess wheelchair-specific anaerobic work capacity?

ObjectiveTo evaluate whether outcomes based on stopwatch time and power output (PO) over a 15 m-overground wheelchair sprint test can be used to assess wheelchair-specific anaerobic work capacity, by studying their relationship with outcomes on a Wingate-based 30 s-wheelchair ergometer sprint (WAnT).MethodsAble-bodied persons (N = 19, 10 men, aged 18–26 y) performed a 15 m overground sprint test in an instrumented wheelchair and a WAnT. 15 m-outcomes were based on stopwatch time (time and mean velocity over 15 m) and on PO (primary outcome: highest mean unilateral PO over successive 5 s-intervals (P5-15m)). WAnT-outcomes were mean unilateral PO over 30 s and the highest mean unilateral PO over successive 5 s-intervals. Correlation coefficients (Pearson's r) and coefficients of determination (R²) were calculated between 15 m-sprint outcomes and WAnT-outcomes.ResultsTime over 15 m (7.2 s (±1.0)) was weakly related to WAnT-outcomes (r = −0.61 and −0.60, R² = 0.38 and 0.36, p < 0.01), similar to mean velocity over 15 m (2.1 m·s⁻¹ (±0.3), R² = 0.43 and 0.39, p < 0.01). P5-15m (38.1 W (±14.0)) showed a moderate relationship to WAnT-outcomes (r = 0.77 and 0.75, R² = 0.59 and 0.56, p < 0.001).ConclusionsIt seems that outcomes based on stopwatch time over a 15 m-overground sprint cannot be used to assess wheelchair-specific anaerobic work capacity, in contrast to an outcome based on PO (P5-15m). The 15 m-sprint with an instrumented wheel can be implemented in rehabilitation practice and research settings when WAnT equipment is not available, although care is needed when interpreting P5-15m as an outcome of anaerobic work capacity given that it seems more skill-dependent than the WAnT.     

TBX6-associated congenital scoliosis (TACS) as a clinically distinguishable subtype of congenital scoliosis: further evidence supporting the compound inheritance and TBX6 gene dosage model

PurposeTo characterize clinically measurable endophenotypes, implicating the TBX6 compound inheritance model.MethodsPatients with congenital scoliosis (CS) from China(N = 345, cohort 1), Japan (N = 142, cohort 2), and the United States (N = 10, cohort 3) were studied. Clinically measurable endophenotypes were compared according to the TBX6 genotypes. A mouse model for Tbx6 compound inheritance (N = 52) was investigated by micro computed tomography (micro-CT). A clinical diagnostic algorithm (TACScore) was developed to assist in clinical recognition of TBX6-associated CS (TACS).ResultsIn cohort 1, TACS patients (N = 33) were significantly younger at onset than the remaining CS patients (P = 0.02), presented with one or more hemivertebrae/butterfly vertebrae (P = 4.9 × 10^‒8), and exhibited vertebral malformations involving the lower part of the spine (T8–S5, P = 4.4 × 10^‒3); observations were confirmed in two replication cohorts. Simple rib anomalies were prevalent in TACS patients (P = 3.1 × 10^‒7), while intraspinal anomalies were uncommon (P = 7.0 × 10^‒7). A clinically usable TACScore was developed with an area under the curve (AUC) of 0.9 (P = 1.6 × 10^‒15). A Tbx6^{-/mh (mild-hypomorphic)} mouse model supported that a gene dosage effect underlies the TACS phenotype.ConclusionTACS is a clinically distinguishable entity with consistent clinically measurable endophenotypes. The type and distribution of vertebral column abnormalities in TBX6/Tbx6 compound inheritance implicate subtle perturbations in gene dosage as a cause of spine developmental birth defects responsible for about 10% of CS.     

Keratoconjunctivitis sicca of human T cell lymphotropic virus type 1 (HTLV-1) infected individuals is associated with high levels of HTLV-1 proviral load

BackgroundA high HTLV-1 proviral load is found in HTLV-1-associated diseases, mainly HAM/TSP. However, the association between proviral load and keratoconjunctivitis sicca (KCS) has not been well established.AimTo verify the association between KCS and HTLV-1 proviral load.Study design104 HTLV-1 infected patients (51 asymptomatic and 52 with HAM/TSP) from the HTLV reference center in Salvador, Brazil were followed from June 2008 to May 2010. Evaluation of tear secretion was performed by BUT (break-up time), Rose Bengal and Schirmer I tests. The diagnosis of KCS was based upon the presence of symptoms and when at least two of three tests were positive. HTLV-1 proviral load was determined using real-time PCR.ResultsThe prevalence of KCS was 44.2%. KCS was more frequent among HAM/TSP patients (p = 0.022). Patients with KCS had higher proviral load (mean 134,672 ± 150,393 copies/10⁶ PBMC) than patients without the disease (mean 66,880 ± 109,525 copies/10⁶ PBMC) (p = 0.001). HTLV-1 proviral load > 100,000 copies/10⁶ PBMC increased significantly the risk of developing KCS (OR = 4.05 and 95% CI = 1.40–11.76). After age > 45 years and HAM/TSP status were excluded in stepway reward analysis, the variables PVL > 100,000 (OR = 4.77 and 95% CI = 1.83–12.44) still remained statistically significant.ConclusionHTLV-1 proviral loads are higher in patients with KCS and may represent a relevant biological marker of disease.     

The role of doxorubicin in non-viral gene transfer in the lung

Proteasome inhibitors have been shown to increase adeno-associated virus (AAV)-mediated transduction in vitro and in vivo. To assess if proteasome inhibitors also increase lipid-mediated gene transfer with relevance to cystic fibrosis (CF), we first assessed the effects of doxorubicin and N-acetyl-l-leucinyl-l-leucinal-l-norleucinal in non-CF (A549) and CF (CFTE29o-) airway epithelial cell lines. CFTE29o- cells did not show a response to Dox or LLnL; however, gene transfer in A549 cells increased in a dose-related fashion (p < 0.05), up to approximately 20-fold respectively at the optimal dose (no treatment: 9.3 × 10⁴ ± 1.5 × 10³, Dox: 1.6 × 10⁶ ± 2.6 × 10⁵, LLnL: 1.9 × 10⁶ ± 3.2 × 10⁵ RLU/mg protein). As Dox is used clinically in cancer chemotherapy we next assessed the effect of this drug on non-viral lung gene transfer in vivo. CF knockout mice were injected intraperitoneally (IP) with Dox (25–100 mg/kg) immediately before nebulisation with plasmid DNA carrying a luciferase reporter gene under the control of a CMV promoter/enhancer (pCIKLux) complexed to the cationic lipid GL67A. Dox also significantly (p < 0.05) increased expression of a plasmid regulated by an elongation factor 1α promoter (hCEFI) approximately 8-fold. Although administration of Dox before lung gene transfer may not be a clinically viable option, understanding how Dox increases lung gene expression may help to shed light on intracellular bottle-necks to gene transfer, and may help to identify other adjuncts that may be more appropriate for use in man.