Corrigendum: Towards automatic pulmonary nodule management in lung cancer screening with deep learning.

This corrects the article DOI: 10.1038/srep46479.

Expansion of spatial and host range of Puumala virus in Sweden: an increasing threat for humans?

Hantaviruses are globally distributed and cause severe human disease. Puumala hantavirus (PUUV) is the most common species in Northern Europe, and the only hantavirus confirmed to circulate in Sweden, restricted to the northern regions of the country. In this study, we aimed to further add to the natural ecology of PUUV in Sweden by investigating prevalence, and spatial and host species infection patterns. Specifically, we wanted to ascertain whether PUUV was present in the natural reservoir, the bank vole (Myodes glareolus) further south than Dalälven river, in south-central Sweden, and whether PUUV can be detected in other rodent species in addition to the natural reservoir. In total, 559 animals were collected at Grimsö (59°43'N; 15°28'E), Sala (59°55'N; 16°36'E) and Bogesund (59°24'N; 18°14'E) in south-central Sweden between May 2013 and November 2014. PUUV ELISA-reactive antibodies were found both in 2013 (22/295) and in 2014 (18/264), and nine samples were confirmed as PUUV-specific by focus reduction neutralization test. Most of the PUUV-specific samples were from the natural host, the bank vole, but also from other rodent hosts, indicating viral spill-over. Finally, we showed that PUUV is present in more highly populated central Sweden.

No evidence for homosubtypic immunity of influenza H3 in Mallards following vaccination in a natural experimental system.

The Mallard (Anas platyrhynchos) is an important reservoir species for influenza A viruses (IAV), and in this host, prevalence and virus diversity are high. Studies have demonstrated the presence of homosubtypic immunity, where individuals are unlikely to be reinfected with the same subtype within an autumn season. Further, evidence for heterosubtypic immunity exists, whereby immune responses specific for one subtype offer partial or complete protection against related HA subtypes. We utilized a natural experimental system to determine whether homo- or heterospecific immunity could be induced following experimental vaccination. Thirty Mallards were vaccinated with an inactivated H3, H6 or a sham vaccine and after seroconversion were exposed to naturally infected wild conspecifics. All ducks were infected within 2 days and had both primary and secondary infections. Overall, there was no observable difference between groups; all individuals were infected with H3 and H10 IAV. At the cessation of the experiment, most individuals had anti-NP antibodies and neutralizing antibodies against H10. Not all individuals had H3 neutralizing antibodies. The isolated H3 IAVs revealed genetic dissimilarity to the H3 vaccine strain, specifically substitutions in the vicinity of the receptor-binding site. There was no evidence of vaccine-induced homosubtypic immunity to H3, a likely result of both a poor H3 immune response in the ducks and H3 immune escape. Likewise, there was no observed heterosubtypic protection related to H6 vaccination. This study highlights the need for experimental approaches to assess how exposure to pathogens and resulting immune processes translates to individual and population disease dynamics.

Priming the immune system of Penaeid shrimp by bacterial HSP70 (DnaK).

This study was conducted to test the effect of DnaK on priming immune responses in Penaeid shrimp. Juvenile-specific pathogen-free (SPF) P. vannamei shrimp were injected with 0.05 µg recombinant DnaK. One hour post-DnaK priming, a non-lethal dose of Vibrio campbellii (10(5) CFU shrimp(-1)) was injected. Other treatments include only DnaK or V. campbellii injection or control with blank inocula. The haemolymph of three shrimp from each treatment was collected at 1.5, 6, 9 and 12 h post-DnaK priming (hpp). It was verified that injection with DnaK and V. campbellii challenge affected the transcription of 3 immune genes, transglutaminase-1 (TGase-1), prophenoloxidase-2 (proPO-2) and endogenous HSP70 (lvHSP70). In P. monodon, shrimp were first injected with DnaK at a dose of 10 µg shrimp(-1) and one hour later with 10(6) CFU of V. harveyi (BB120) shrimp(-1). Shrimp injected with DnaK showed a significant increase in proPO expression compared to the control (P < 0.05). Yet a double injection (DnaK and Vibrio) seemed to cause an antagonistic response at the level of expression, which was not equalled at the level of PO activity. Those results suggest that DnaK is able to modulate immune responses in P. vannamei and P. monodon.

[Calcaneo-fibular ligament surgery for chronic lateral instability of the upper ankle : Broström technique with modification by Wille. Video article]. / Fibulare Bandplastik bei chronischer lateraler Instabilität am oberen Sprunggelenk : Operationstechnik nach Broström in der Modifikation nach Wille. Videobeitrag.

BACKGROUND: An acute ligament rupture of the lateral ligament complex of the ankle joint is treated without surgery. Treatment failure may lead to a chronically unstable situation of the ankle joint, in which case surgery is an effective procedure for stabilizing the ruptured ligaments. Anatomical reconstruction is the best operative technique if the ligament tissue is of good quality. METHOD: In our video we demonstrate a new possibility for the positioning of an anchor to tighten the calcaneo-fibular ligament. Modified Broström repairs are described in the literature in which the calcaneo-fibular ligament is released and reattached to the fibula to tighten it. We present the option to release the ligament at the calcaneus and reattach it using a suture anchor. This offers the advantage of preventing the possible dislocation of the peroneal tendons.

Comparison of active-set method deconvolution and matched-filtering for derivation of an ultrasound transit time spectrum.

The quality of ultrasound computed tomography imaging is primarily determined by the accuracy of ultrasound transit time measurement. A major problem in analysis is the overlap of signals making it difficult to detect the correct transit time. The current standard is to apply a matched-filtering approach to the input and output signals. This study compares the matched-filtering technique with active set deconvolution to derive a transit time spectrum from a coded excitation chirp signal and the measured output signal. The ultrasound wave travels in a direct and a reflected path to the receiver, resulting in an overlap in the recorded output signal. The matched-filtering and deconvolution techniques were applied to determine the transit times associated with the two signal paths. Both techniques were able to detect the two different transit times; while matched-filtering has a better accuracy (0.13 µs versus 0.18 µs standard deviations), deconvolution has a 3.5 times improved side-lobe to main-lobe ratio. A higher side-lobe suppression is important to further improve image fidelity. These results suggest that a future combination of both techniques would provide improved signal detection and hence improved image fidelity.

Effects of exercise and hypoxia on heart rate variability and acute mountain sickness.

Acute mountain sickness (AMS) is a common condition among non-acclimatized individuals ascending to high altitude. Exercise, a characteristic feature of hiking and mountaineering, has been suggested to exacerbate AMS prevalence and to cause modifications of the autonomic nervous system. A reduction of the heart rate variability (HRV) is a common finding during acute hypoxia, however characteristics of HRV during exercise in subjects suffering from AMS are unknown. Therefore, the aim of the present study was to investigate the effects of acute normobaric hypoxia (FiO2=11.0% ≙ 5 500 m) at rest (PHE) and during exercise (AHE) on the cardiac autonomic function and the development of AMS in 20 healthy, male individuals. HRV recordings were performed during normoxia and after 2, 4, 6 and 8 h in hypoxia during PHE and AHE, respectively. AMS was assessed using the Lake Louise Score. During PHE 50% of participants developed AMS and 70% during AHE (p=0.22). The analysis of HRV data showed a significant reduction of total power (TP), high frequency (HF) and low frequency (LF) components and an increase of the LF:HF ratio during PHE, however without further modification during AHE. Exercise did not increase AMS prevalence or severity, but increased "non-gastrointestinal" symptoms including headache, fatigue and dizziness. HRV indices were not related to the overall incidence of AMS or the development of "non-gastrointestinal" symptoms but we detected significant correlations between gastrointestinal complaints and HRV components. Thus, we suggest that the cardiac autonomic modulation during acute normobaric hypoxia does not play an important role in the development of AMS, but seems to be related to gastrointestinal complaints at high altitude. However, the influence of moderate exercise on HRV and AMS is minor, only "non-GI" symptoms seem to be exacerbated when exercise is applied.

Effects of massage under hypoxic conditions on exercise-induced muscle damage and physical strain indices in professional soccer players.

Reports based on experiences from masseurs and players, mostly without any scientific background, suggest that the combination of a classical regeneration method (i.e. massage) with exposure to hypoxia may enhance regeneration in soccer. The aim of this study was to evaluate whether this specific combination could affect blood parameters related to muscle damage and physical strain after a soccer game. Approximately 15 hours after two separate championship games, 10 professional male outfield players of the first Austrian division were exposed to normobaric hypoxia (FiO2 13.5% ∼ 4000m) or normoxia for 1 hour (30 minutes rest followed by 30 min massage) (cross-over design). Creatine kinase (CK), urea and uric acid (UA) were measured 4 days before the first game, and 15 and 63 hours after the two games. Match play increased CK values independently of the intervention. No effect of the massage in combination with hypoxia was seen. A trend was found between Δ UA ([UA] 48 hours after exposure minus [UA] before exposure) in response to hypoxia and SaO2 measured in hypoxia (r=0.612, p=0.06). Results show that massage under hypoxic conditions had no additional positive effect on the measured parameters compared to massage alone. Solely the trend of a relationship for Δ UA and SaO2 might indicate that redox alterations are a potential consequence of hypoxic exposure.

An inexpensive and effective larval diet for Anopheles arabiensis (Diptera: Culicidae): eat like a horse, a bird, or a fish?

A successful sterile insect technique program depends upon mass production of good-quality sterile insects for release into a target area. Specifically, to control Anopheles arabiensis Patton (Diptera: Culicidae) in a pilot area in northern Sudan, 1 million An. arabiensis sterile males per day are needed. To reach this production objective, mosquito mass rearing is indispensable and larval diet, a key parameter for the production of healthy male mosquitoes, needs to be cost-effective. The Koi Floating Blend fish food, previously used at the Food and Agriculture Organization/International Atomic Energy Agency Insect Pest Control Laboratory for the routine rearing of the An. arabiensis colony, is no longer available. The aim of this study was to find a cheap and effective substitute for the discontinued diet. Several candidate powdered diets that are commercially available were tested to determine the best diet. By using mixture experiment principles and response surface methodology, the combination of two components, bovine liver powder and tuna meal, showed the best results in terms of larval survival, developmental, rate and adult size. The addition of a vitamin mixture further improved the diet. These positive production results coupled with the relatively low cost of our blend demonstrated the possibility of its use for mass rearing purpose.

Short-term intermittent hypoxia reduces the severity of acute mountain sickness.

Intermittent hypoxia (IH) is a promising approach to induce acclimatization and hence lower the risk of developing acute mountain sickness (AMS). We hypothesized that a short-term IH protocol in normobaric hypoxia (7 × 1 h to 4500 m) effectively increases the hypoxic ventilatory response (HVR) and reduces the incidence and severity of AMS. Therefore, 26 men (25.5 ± 4.4 years), assigned in a double-blinded fashion to the hypoxia group (HG) or placebo group (PG), spent 8 h at 5300 m before (PRE) and 2 days after cessation of the IH protocol (POST). Measurements included the evaluation of the Lake Louise Score (LLS) and the HVR. The severity of AMS decreased from PRE to POST in the HG (from 6.0 ± 2.7 at PRE to 4.1 ± 2.1 at POST), whereas the LLS in the PG stayed high (from 5.7 ± 2.9 to 5.5 ± 2.8, respectively). The HVR in the HG increased from 0.73 ± 0.4 L/min/% at PRE to 1.10 ± 0.5 L/min/% at POST and did not increase in the PG. The reduction of the LLS was inversely related to the changes in the HVR (r = -0.434), but the AMS incidence was not different between the HG and the PG at POST. In conclusion, short-term IH reduced the severity of AMS development during a subsequent 8-h exposure to normobaric hypoxia.

Effect of poly-ß-hydroxybutyrate on Chinese mitten crab, Eriocheir sinensis, larvae challenged with pathogenic Vibrio anguillarum.

This study investigated the protective effect of poly-ß-hydroxybutyrate (PHB) on Chinese mitten crab, Eriocheir sinensis, zoea larvae challenged with pathogenic Vibrio anguillarum. PHB was delivered to the crab larvae through rotifer and Artemia bioencapsulation. Zoea 3 larvae were challenged with V. anguillarum at a final concentration of 10(5) CFU mL(-1). PHB-enriched rotifers and Artemia nauplii were added to the culture water 24 h prior to, upon and 24 h after challenge. The results confirmed that PHB could enhance the survival and growth of unexposed E. sinensis larvae. Moreover, PHB protected larvae from the pathogen as the larvae fed PHB-enriched live food showed the highest survival and development rate in all challenged groups (P < 0.05). Furthermore, larval performance was the best when PHB was delivered to the larvae 24 h before challenge (P < 0.05). In conclusion, our results indicate that PHB can be used as part of an effective strategy to protect E. sinensis larvae from V. anguillarum resulting in higher survival and better growth, especially when applied before the challenge.

Changes in cardiac autonomic activity during a passive 8 hour acute exposure to 5 500 m normobaric hypoxia are not related to the development of acute mountain sickness.

Alterations in the autonomic nervous system after ascent to high altitude may be related to the development of acute mountain sickness (AMS). So far, the time course of cardiac autonomic modulation in relation to AMS development during the early hours at altitude is not well established. As AMS develops sometimes as early as 1 h and typically within 6 to 10 h at altitude, evaluating this time period provides information on cardiac autonomic responses with regard to AMS development. Prior studies exclusively investigated autonomic modulations in hypobaric hypoxia. Because barometric pressure per se might influence autonomic nervous system activity, the evaluation of cardiac autonomic alterations caused by hypoxia alone might give new insights on the role of the autonomic nervous system in AMS development. To assess the early responses of acute hypoxia on cardiac autonomic modulation and its association to the development of AMS, 48 male subjects were exposed for 8 h to acute normobaric hypoxia (FiO2 11.0%, 5 500 m respectively). Heart rate variability (HRV) was determined by 5-min recordings of successive NN-intervals in normoxia and after 2, 4, 6 and 8 h in hypoxia. Compared with normoxia, acute exposure to hypoxia decreased total power (TP), high frequency (HF) and low frequency (LF) components as well as the standard deviation of all NN intervals (SDNN), the root mean square of differences of successive NN intervals (rMSSD) and the proportion of differences between adjacent NN intervals of more than 50 ms (pNN50). LF:HF ratio, heart rate (HR) and blood lactate (LA) were augmented, indicating an increase in cardiac sympathetic activity. No differences were found between those who developed AMS and those who did not. Our results confirm reduced HRV with a shift towards sympathetic predominance during acute exposure to hypoxia. However, changes in cardiac autonomic modulations are not related to AMS development in acute normobaric hypoxia.

[Tibiocalcaneal arthrodesis using retrograde insertion of a compression nail]. / Die tibiokalkaneare Arthrodese mit retrograd eingebrachtem Kompressionsnagel.

UNLABELLED: OPERATION GOAL: Arthrodesis of the upper and lower ankle joint because of problematic bone positioning or failed arthrodesis. Osteosynthesis procedure using a retrograde compression nail. To achieve stable, fully weight-bearing osteosynthesis for early, pain-free mobilization. INDICATIONS: Rearthrodesis because of failure of the conventional arthrodesis technique and development of osteoarthritis of the lower ankle joint. Painful osteoarthritis of the upper ankle joint because of inadequate perfusion or a major bone defect because of sclerosis or necrosis. Primary arthrodesis because of facture of the lower leg (pilon tibial) with joint involvement and preexisting osteoarthritis. CONTRAINDICATIONS: Acute osteitis/osteomyelitis, sclerosis in the marrow of the distal tibia, malalignment of the distal tibial shaft and local soft tissue inflammation. SURGICAL TECHNIQUE: Preparation of the articular surface of the upper and lower ankle for arthrodesis using a transfibular approach. If necessary, correction of bone defects with iliac crest spongiosa. Stabile osteosynthesis by retrograde insertion of a compression nail. POSTOPERATIVE MANAGEMENT: A split lower leg cast on the 2nd postoperative day, mobilization of the patient with underarm crutches with floor contact for 2 weeks, then with application of a lower leg walking cast for 8 weeks with partial weight-bearing for 4 weeks and full weight-bearing for the last 4 weeks of cast fixation. X-ray controls immediately postoperatively, then after 6 and 12 weeks. RESULTS: From 2006 to 2008, 12 patients (7 men, 5 women; mean age 59 years) with various indications were treated with retrograde insertion of a compression nail. All patients were routinely controlled radiologically and clinically after 2, 4, 8 and 12 weeks. Follow-up was carried out at 6, 12 and 24 months. All arthrodeses showed osseous consolidation 16 weeks postoperatively. Ten patients were able to use full weight-bearing without pain after 12 weeks. Two patients reported experiencing pain after walking for 2 h. In total three complications occurred: one hindfoot healed with varus malalignment; one patient fell, fracturing the lower leg above the nail; one distal locking screw loosened.

Quorum quenching bacteria protect Macrobrachium rosenbergii larvae from Vibrio harveyi infection.

AIMS: In this study, we investigated the effect of N-acyl homoserine lactone-degrading bacterial enrichment cultures (ECs) on larviculture of the giant freshwater prawn Macrobrachium rosenbergii. METHODS AND RESULTS: The larval performance in terms of larval growth, larval survival, larval quality, duration of the larval rearing process and microflora levels in the rearing water as well as inside the prawn gut was investigated. The application of the EC bacteria was performed in two ways: by adding them directly into the larval rearing water and via enriched Artemia nauplii used for larval feeding. The results of the study demonstrated that both ECs that were tested had a similar positive effect on larval survival and larval quality, whereas they did not affect larval growth or the duration of the larval rearing process. CONCLUSIONS: Under normal hatchery conditions, the optimal EC densities were found to be 10(6) CFU ml(-1) for adding into the rearing water and 5 × 10(8) CFU ml(-1) for enrichment of Artemia nauplii used for feeding of the larvae. In the hatchery, the ECs can be grown on waste streams of Artemia hatching. SIGNIFICANCE AND IMPACT OF THE STUDY: Application of this kind of ECs could lead to a more sustainable aquaculture production, by replacing the use of antibiotics to control diseases.

Virulence of white spot syndrome virus (WSSV) isolates may be correlated with the degree of replication in gills of Penaeus vannamei juveniles.

A standardized inoculation model was used in 2 separate experiments to gauge the virulence of 3 white spot syndrome virus (WSSV) isolates from Thailand and Vietnam (WSSV Thai-1, WSSV Thai-2, and WSSV Viet) in Penaeus vannamei juveniles. Mortality patterns (Expt 1) were compared and WSSV-positive cells quantified (Expt 2) in tissues following intramuscular inoculation of shrimp with the most (WSSV Thai-1) and least (WSSV Viet) virulent isolates as determined by Expt 1. The results of Expt 1 demonstrated that mortalities began at 36 h post inoculation (hpi) for both Thai isolate groups and at 36 to 60 hpi for the Viet isolate group. Cumulative mortality reached 100% 96 to 240 h later in shrimp challenged with the WSSV Viet isolate compared to shrimp challenged with the Thai isolates. WSSV infection was verified in all groups by indirect immunofluorescence. In Expt 2, WSSV-infected cells were quantified by immunohistochemical analysis of both dead and time-course sampled shrimp. WSSV-positive cells were detected in tissues of Thai-1 inoculated dead and euthanized shrimp from 24 hpi onwards and from 36 hpi onwards in shrimp injected with the Viet isolate. Significantly more infected cells were found in tissues of dead shrimp inoculated with the Thai-1 than in Viet isolate-inoculated shrimp. In these experiments, substantial differences in virulence were demonstrated between the WSSV isolates. The Vietnamese isolate induced a more chronic disease and mortality pattern than was found for the Thai isolates, possibly because it infected fewer cells. This difference was most pronounced in gills.

A review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus.

Since it first appeared in 1992, white spot syndrome virus (WSSV) has become the most threatening infectious agent in shrimp aquaculture. Within a decade, this pathogen has spread to all the main shrimp farming areas and has caused enormous economic losses amounting to more than seven billion US dollars. At present, biosecurity methods used to exclude pathogens in shrimp farms include disinfecting ponds and water, preventing the entrance of animals that may carry infectious agents and stocking ponds with specific pathogen-free post-larvae. The combination of these practices increases biosecurity in shrimp farming facilities and may contribute to reduce the risk of a WSSV outbreak. Although several control methods have shown some efficacy against WSSV under experimental conditions, no therapeutic products or strategies are available to effectively control WSSV in the field. Furthermore, differences in virulence and clinical outcome of WSSV infections have been reported. The sequencing and characterization of different strains of WSSV has begun to determine aspects of its biology, virulence and pathogenesis. Knowledge on these aspects is critical for developing effective control methods. The aim of this review is to present an update of the knowledge generated so far on different aspects of WSSV organization, morphogenesis, pathology and pathogenesis.

Pathogenesis of a Thai strain of white spot syndrome virus (WSSV) in juvenile, specific pathogen-free Litopenaeus vannamei.

White spot syndrome virus (WSSV) causes disease and mortality in cultured and wild shrimp. A standardized WSSV oral inoculation procedure was used in specific pathogen-free (SPF) Litopenaeus vannamei (also called Penaeus vannamei) to determine the primary sites of replication (portal of entry), to analyze the viral spread and to propose the cause of death. Shrimp were inoculated orally with a low (10(1.5) shrimp infectious dose 50% endpoint [SID50]) or a high (10(4) SID50) dose. Per dose, 6 shrimp were collected at 0, 6, 12, 18, 24, 36, 48 and 60 h post inoculation (hpi). WSSV-infected cells were located in tissues by immunohistochemistry and in hemolymph by indirect immunofluorescence. Cell-free hemolymph was examined for WSSV DNA using 1-step PCR. Tissues and cell-free hemolymph were first positive at 18 hpi (low dose) or at 12 hpi (high dose). With the 2 doses, primary replication was found in cells of the foregut and gills. The antennal gland was an additional primary replication site at the high dose. WSSV-infected cells were found in the hemolymph starting from 36 hpi. At 60 hpi, the percentage of WSSV-infected cells was 36 for the epithelial cells of the foregut and 27 for the epithelial cells of the integument; the number of WSSV-infected cells per mm2 was 98 for the gills, 26 for the antennal gland, 78 for the hematopoietic tissue and 49 for the lymphoid organ. Areas of necrosis were observed in infected tissues starting from 48 hpi (low dose) or 36 hpi (high dose). Since the foregut, gills, antennal gland and integument are essential for the maintenance of shrimp homeostasis, it is likely that WSSV infection leads to death due to their dysfunction.

[Inner amputation of an upper extremity with impaired cerebral perfusion and lethal outcome]. / Innere Amputation einer oberen Extremität mit zerebraler Perfusionsstörung und letalem Ausgang.

An inner amputation of the upper extremity is understood to be a rupture of the arm's vascular nerve bundle as well as the shoulder's and scapular's stabilizing muscles and fibrous joints without damage to the dermal soft tissue sheath. This injury is a rare and grave incident which mostly occurs within the scope of a high-energy trauma and in cases of polytraumatized patients in combination with additional life-threatening injuries. In the literature this is referred to by the terms scapulothoracic dissociation (SD) and closed forequarter amputation--the entity of SD was first described by Oreck et al. in 1984. Traction of the upper extremity leads to the rupture of the m. pectoralis major, m. pectoralis minor, the musculi rhomboidei, the m. levator scapulae, the m. latissimus dorsi as well as the m. trapezius. Furthermore the traction causes acromioclavicular or sternoclavicular bursting of the joints or a displaced clavicle fracture. Among the more than 50 cases depicted until now, 94% of the patients exhibit a neurological (plexus) and 80% a vascular lesion, thus corresponding to a genuine inner amputation. The prognosis for this injury is consistently poor: 10% of the patients die, in 52% an nonfunctional extremity remains, and in 21% a untimely amputation has to be performed. The observed complication of cerebral hypoperfusion caused by increasing pressure in the neck compartment, which ultimately leads to the death of the patient, has, as far as we know, not yet been specified and emphasizes the gravity and the magnitude as well as the necessity of rapid diagnosis and appropriate therapy of this infrequent injury.