Are the common sterilization methods completely effective for our in-house 3D printed biomodels and surgical guides?

INTRODUCTION: In-hospital 3D printing is being implemented in orthopaedic departments worldwide, being used for additive manufacturing of fracture models (or even surgical guides) which are sterilized and used in the operating room. However, to save time and material, prints are nearly hollow, while 3D printers are placed in non-sterile rooms. The aim of our study is to evaluate whether common sterilization methods can sterilize the inside of the pieces, which would be of utmost importance in case a model breaks during a surgical intervention. MATERIAL AND METHOD: A total of 24 cylinders were designed and printed with a 3D printer in Polylactic Acid (PLA) with an infill density of 12%. Manufacturing was paused when 60% of the print was reached and 20 of the cylinders were inoculated with 0.4 mL of a suspension of S epidermidis ATTCC 1228 in saline solution at turbidity 1 McFarland. Printing was resumed, being all the pieces completely sealed with the inoculum inside. Posteriorly, 4 groups were made according to the chosen sterilization method: Ethylene Oxide (EtO), Gas Plasma, Steam Heat or non-sterilized (positive control). Each group included 5 contaminated cylinders and 1 non-contaminated cylinder as a negative control. After sterilization, the inside of the cylinders was cultured during 7 days. RESULTS: We observed bacterial growth of just a few Forming Colony Units (FCU) in 4 out of 5 positive controls and in 2 out of 5 contaminated cylinders sterilized with Gas Plasma. We could not assess any bacterial growth in any of the EtO or Steam Heat samples or in any of the negative controls. Pieces sterilized under Steam Heat resulted completely deformed. CONCLUSIONS: High temperatures reached during the procedure of additive manufacturing can decrease the bacterial load of the biomodels. However, there is a potential risk of contamination during the procedure. We recommend sterilization with EtO for in-hospital 3D-printed PLA hollow biomodels or guides. Otherwise, in case of using Gas Plasma, an infill of 100% should be applied.

A dietary intervention with functional foods reduces metabolic endotoxaemia and attenuates biochemical abnormalities by modifying faecal microbiota in people with type 2 diabetes.

AIM: To study the effects of a functional food-based dietary intervention on faecal microbiota and biochemical parameters in patients with type 2 diabetes (T2D). MATERIALS AND METHODS: This placebo-controlled, randomized, double-blind study included 81 patients with T2D divided into two 3-month treatment groups: one following a reduced-energy diet with a dietary portfolio (DP) comprising high-fibre, polyphenol-rich and vegetable-protein functional foods; the other taking a placebo (P). The primary outcome was the effect of the DP on faecal microbiota. Secondary endpoints were biochemical parameters, lipopolysaccharide, branched-chain amino acids, trimethylamine N-oxide, glycosylated haemoglobin (HbA1c) and free fatty acids (FFAs). RESULTS: Patients with T2D exhibited intestinal dysbiosis characterized by an increase in Prevotella copri. Dietary intervention with functional foods significantly modified faecal microbiota compared with P by increasing alpha diversity and modifying the abundance of specific bacteria, independently of antidiabetic drugs. There was a decrease in P. copri and increases in Faecalibacterium prausnitzii and Akkermansia muciniphila, two bacterial species known to have anti-inflammatory effects. The DP group also exhibited significant reductions in areas under the curve for glucose, total and LDL cholesterol, FFAs, HbA1c (P< 0.05), triglycerides and CRP, and an increase in antioxidant activity (P< 0.01) vs. the P group. CONCLUSION: Long-term adherence to a high-fibre, polyphenol-enriched and vegetable-protein-based diet provides benefits for the composition of faecal microbiota, and may offer potential therapies for improvement of glycaemic control, dyslipidaemia and inflammation.

Length-scale dependency of biomimetic hard-soft composites.

Biomimetic composites are usually made by combining hard and soft phases using, for example, multi-material additive manufacturing (AM). Like other fabrication methods, AM techniques are limited by the resolution of the device, hence, setting a minimum length scale. The effects of this length scale on the performance of hard-soft composites are not well understood. Here, we studied how this length scale affects the fracture toughness behavior of single-edge notched specimens made using random, semi-random, and ordered arrangements of the hard and soft phases with five different ratios of hard to soft phases. Increase in the length scale (40 to 960 µm) was found to cause a four-fold drop in the fracture toughness. The effects of the length scale were also modulated by the arrangement and volumetric ratio of both phases. A decreased size of the crack tip plastic zone, a crack path going through the soft phase, and highly strained areas far from the crack tip were the main mechanisms explaining the drop of the fracture toughness with the length scale.

Pollen Types Used by the Native Stingless Bee, Tetragonisca angustula (Latreille), in an Amazon-Chiquitano Transitional Forest of Bolivia.

Samples of corbicular and stored pollen gathered by Tetragonisca angustula (Latreille, 1811) in an Amazon-Chiquitano transitional forest during the dry season were analyzed. The pollen spectrum was established as well as the dynamics of the relationship between the stingless bee and the surrounding flora. Pollen samples obtained from three rational hives were subjected to acetolysis and 55 pollen types were identified, the most frequent being from Anadenanthera (Fabaceae), Chenopodiaceae, and Dydimopanax (Araliaceae). Significant differences in pollen families used between hives along the months of collection were found in stored pollen and non-significant differences in corbicular pollen. Mean values of alpha diversity (H') showed T. angustula as a generalist while beta diversity qualitatively showed that pollen composition was similar between two hives both of which differed from the third hive. Pollen types in corbicular and stored pollen were in general related with richness of flowering plants in the foraging area; the pollen offer was less diverse than the pollen collected, and a trend to prefer the collection of pollen from the most abundant flowering species could be discerned. T. angustula showed polylectic feeding habits and a capacity to adapt to food availability.

Avatars and virtual agents - relationship interfaces for the elderly.

In the Digital Era, the authors witness a change in the relationship between the patient and the care-giver or Health Maintenance Organization's providing the health services. Another fact is the use of various technologies to increase the effectiveness and quality of health services across all primary and secondary users. These technologies range from telemedicine systems, decision making tools, online and self-services applications and virtual agents; all providing information and assistance. The common thread between all these digital implementations, is they all require human machine interfaces. These interfaces must be interactive, user friendly and inviting, to create user involvement and cooperation incentives. The challenge is to design interfaces which will best fit the target users and enable smooth interaction especially, for the elderly users. Avatars and Virtual Agents are one of the interfaces used for both home care monitoring and companionship. They are also inherently multimodal in nature and allow an intimate relation between the elderly users and the Avatar. This study discusses the need and nature of these relationship models, the challenges of designing for the elderly. The study proposes key features for the design and evaluation in the area of assistive applications using Avatar and Virtual agents for the elderly users.

Transcriptome Analysis of Targeted Mouse Mutations Reveals the Topography of Local Changes in Gene Expression.

The unintended consequences of gene targeting in mouse models have not been thoroughly studied and a more systematic analysis is needed to understand the frequency and characteristics of off-target effects. Using RNA-seq, we evaluated targeted and neighboring gene expression in tissues from 44 homozygous mutants compared with C57BL/6N control mice. Two allele types were evaluated: 15 targeted trap mutations (TRAP); and 29 deletion alleles (DEL), usually a deletion between the translational start and the 3' UTR. Both targeting strategies insert a bacterial beta-galactosidase reporter (LacZ) and a neomycin resistance selection cassette. Evaluating transcription of genes in +/- 500 kb of flanking DNA around the targeted gene, we found up-regulated genes more frequently around DEL compared with TRAP alleles, however the frequency of alleles with local down-regulated genes flanking DEL and TRAP targets was similar. Down-regulated genes around both DEL and TRAP targets were found at a higher frequency than expected from a genome-wide survey. However, only around DEL targets were up-regulated genes found with a significantly higher frequency compared with genome-wide sampling. Transcriptome analysis confirms targeting in 97% of DEL alleles, but in only 47% of TRAP alleles probably due to non-functional splice variants, and some splicing around the gene trap. Local effects on gene expression are likely due to a number of factors including compensatory regulation, loss or disruption of intragenic regulatory elements, the exogenous promoter in the neo selection cassette, removal of insulating DNA in the DEL mutants, and local silencing due to disruption of normal chromatin organization or presence of exogenous DNA. An understanding of local position effects is important for understanding and interpreting any phenotype attributed to targeted gene mutations, or to spontaneous indels.

Reporter Gene Silencing in Targeted Mouse Mutants Is Associated with Promoter CpG Island Methylation.

Targeted mutations in mouse disrupt local chromatin structure and may lead to unanticipated local effects. We evaluated targeted gene promoter silencing in a group of six mutants carrying the tm1a Knockout Mouse Project allele containing both a LacZ reporter gene driven by the native promoter and a neo selection cassette. Messenger RNA levels of the reporter gene and targeted gene were assessed by qRT-PCR, and methylation of the promoter CpG islands and LacZ coding sequence were evaluated by sequencing of bisulfite-treated DNA. Mutants were stratified by LacZ staining into presumed Silenced and Expressed reporter genes. Silenced mutants had reduced relative quantities LacZ mRNA and greater CpG Island methylation compared with the Expressed mutant group. Within the silenced group, LacZ coding sequence methylation was significantly and positively correlated with CpG Island methylation, while promoter CpG methylation was only weakly correlated with LacZ gene mRNA. The results support the conclusion that there is promoter silencing in a subset of mutants carrying the tm1a allele. The features of targeted genes which promote local silencing when targeted remain unknown.

Optimal medical, rehabilitation and behavioral management in the setting of failed back surgery syndrome.

INTRODUCTION: Failed back surgery syndrome (FBSS) constitutes a constellation of symptoms grouped together and attributed to prior surgical intervention. Clinicians often poorly understand the heterogeneity of this condition combined with the etiological factors responsible for its development. Therefore, it would follow that an algorithmic treatment approach to patients diagnosed with this syndrome might pose challenges. The clinical work-up of the patient involves history, examination and appropriate diagnostic imaging as well as behavioral assessment. MATERIALS AND METHODS: We sought to conduct a narrative review of the available literature focused on the medical, rehabilitative and behavioral treatment of FBSS. To that end, we conducted a literature search using PubMed (through March 2013). We focused on studies published over the last 20years. Only English language articles were included. Search terms included "failed back surgery syndrome", "FBSS", "failed back syndrome", and "post-laminectomy syndrome". Studies included in our review focused on randomized controlled trials (RCTs), prospective cohort studies, and case series (retrospective and prospective). Studies were organized by intervention (e.g. medical management, physical therapy and rehabilitation, and psychosocial) and presented to emphasize the quality of evidence (e.g. RCTs, prospective studies, etc.). CONCLUSION: Overall, the literature provides very limited guidance on the comprehensive management of patients suffering from FBSS. There are rehabilitative interventions and behavioral protocols that demonstrate promise. Pathways based on medication management remain difficult to clearly define.

Productive HIV-1 infection of human cervical tissue ex vivo is associated with the secretory phase of the menstrual cycle.

Cervical tissue explants (CTEs) from 22 HIV-1 seronegative women were exposed to R5 HIV-1 ex vivo. Eight CTEs were productively infected in terms of HIV-1 p24Gag release in culture supernatants, whereas 14 were not. Nonetheless, both accumulation of HIV-1gag DNA and of p24Gag(+) CD4(+) T cells and macrophages occurred in both productive and, at lower levels, in nonproductive CTEs. Nonproductive CTEs differed from productive CTEs for higher secretion of C-C motif chemokine ligand 3 (CCL3) and CCL5. A post-hoc analysis revealed that all productive CTEs were established from women in their secretory phase of the menstrual cycle, whereas nonproductive CTEs were derived from women either in their secretory (28%) or proliferative (36%) menstrual cycle phases or with an atrophic endometrium (36%). Thus, our results support the epidemiological observation that sexual HIV-1 transmission from males to women as well as from women to men is more efficient during their secretory phase of the menstrual cycle.

Characterization and distribution of tomato yellow margin leaf curl virus, a begomovirus from Venezuela.

A begomovirus causing mottling and leaf deformation in tomato from the State of Mérida was cloned and sequenced. The virus has a bipartite genome comprised of a DNA-A (2,572 nucleotides) and a DNA-B (2,543 nucleotides) with a genome organization typical of New World begomoviruses. Both components share a common region of 115 nucleotides with 98 % sequence identity. Phylogenetic analysis indicated that while no virus sequences were closely related, the A component was distantly related to those of two other tomato-infecting viruses, tomato leaf deformation virus and Merremia mosaic virus; and the DNA-B, to those of pepper huasteco yellow vein virus and Rhynchosia golden mosaic Yucatan virus. The DNA-A and DNA-B sequences were submitted to GenBank (accession no. AY508993 and AY508994, respectively) and later accepted by the International Committee on Taxonomy of Viruses as the genome of a member of a unique virus species with the name Tomato yellow margin leaf curl virus (TYMLCV). Tomato (Solanum lycopersicum L. 'Fl. Lanai') plants inoculated with cloned TYMLCV DNA-A and DNA-B became systemically infected and showed chlorotic margins and leaf curling. The distribution of TYMLCV in tomato-producing states in Venezuela was determined by nucleic acid spot hybridization analysis of 334 tomato leaf samples collected from ten states using a TYMLCV-specific probe and confirmed by PCR and sequencing of the PCR fragment. TYMLCV was detected in samples from the states of Aragua, Guárico, and Mérida, suggesting that TYMLCV is widely distributed in Venezuela.

Piracetam-induced changes on the brainstem auditory response in anesthetized juvenile rhesus monkeys (Macaca mulatta). Report of two clinical cases.

BACKGROUND: We describe two clinical cases and examine the effects of piracetam on the brainstem auditory response in infantile female rhesus monkeys (Macaca mulatta). RESULTS: We found that the interwave intervals show a greater reduction in a 3-year-old rhesus monkey compared to a 1-year-old rhesus monkey. DISCUSSION: In this report, we discuss the significance of these observations.

The selective inhibition of the D1 dopamine receptor results in an increase of metabolized dopamine in the rat striatum.

Our aim was to study the specific role of the postsynaptic D(1) receptors on dopaminergic response and analyze the metabolized dopamine (DA) in the rat striatum. We used male Wistar rats to evaluate the effects of different doses of a D(1) agonist (SKF-38393) and a D(1) antagonist (SCH-23390), and their co-administration. The levels of DA and L-3, 4-dihydroxyphenylacetic acid (DOPAC) were measured using high performance liquid chromatography. The systemic injection of SKF-38393 alone at 1, 5 and 10 mg/kg did not alter the DA and DOPAC levels or the DOPAC/DA ratio. In contrast, injection of SCH-23390 alone at 0.25, 0.5 and 1 mg/kg significantly increased the DA and DOPAC levels, as well as the DOPAC/DA ratio, compared with the respective control groups. The co-administration of SCH-23390+SKF-38393 did not alter the DA or DOPAC levels, but it did significantly inhibit the SCH-23390-induced increase of the DA and DOPAC levels. The SCH-23390+SKF-38393 and the SCH-23390-only groups showed an increase in the DOPAC/DA ratio. The co-administration of SCH-23390+PARGYLINE significantly decreased the DOPAC levels and the DOPAC/DA ratio compared with the control and SCH-23390 groups. Taken together, our results showed that selective inhibition with SCH-23390 produced an increase in metabolized DA via striatal monoamine oxidase. These findings also contribute to the understanding of the role of postsynaptic D(1) receptors in the long-loop negative feedback system in the rat striatum.

[Paradigm of negative feedback via long-loop in the striatal dopamine release modulation in the rat]. / Paradigma de retroalimentacion negativa via circuito largo en la modulacion de la liberacion de dopamina en el estriado dorsal de la rata.

INTRODUCTION: The basal ganglia include the striatum, globus pallidus, the substantia nigra pars compacta and pars reticulata. The striatum receives afferent input from the substantia nigra pars compacta. The principal neurons of the striatum are medium spiny neurons, that express high levels of D1 and D2 receptors. AIMS: This review deals about the aspects underlying to the negative feedback via long-loop in the striatal dopamine release modulation in the rat. Also, the motor function in dopamine receptor knock-out mice is discussed. DEVELOPMENT AND CONCLUSIONS: The intrastriatal infusion and systemic injection of dopamine receptor agonists and antagonists may regulate the striatal dopamine release and induce changes in motor function. Disruption of the D1 and D2 gene shown that the motor function is controlled by D1 and D2 receptors. The study of the long-loop negative feedback may contribute to our understanding in the physiology and dysfunction of basal ganglia.

Detection of hantaviruses in Brazilian rodents by SYBR-Green-based real-time RT-PCR.

Current knowledge of the pathogenic hantavirus indicates that wild rodents are its primary natural reservoir. Specific primers to detect the presence of viral genomes were developed using an SYBR-Green-based real-time RT-PCR protocol. One hundred sixty-four rodents native to the Atlantic Forest biome were captured in São Paulo State, Brazil, and their tissues were tested. The presence of hantavirus RNA was detected in sixteen rodents: three specimens of Akodon montensis, three of Akodon cursor, two of Necromys lasiurus, one of Juliomys sp., one of Thaptomys nigrita, five of Oligoryzomys nigripes, and one of Oryzomys sp. This SYBR Green real-time RT-PCR method for detection of hantavirus may be useful for surveying hantaviruses in Brazil.

[Health status and life style in Castel Malnome (Rome, I-II cent. A. D.)]. / Attività lavorative e condizioni di vita della comunità di Castel Malnome (Roma, I-II sec. D.C.).

The necropolis of Castel Malnome, chronologically framed between the I and II century AD, is located in the vicinity of Ponte Galeria (Rome), nearby the via Portuense. The excavation of the funerary site has allowed the collection of 292 inhumations, referred to the lower social classes and for the most part adult males. Regarding the funerary ritual, only the 42.8% of the graves had a tiles cover, while about one third provided grave goods. The field analysis shows that almost all the burials are primaries, and is not possible to highlight a main position of the inhumated individuals. The laboratory analysis, till today carried out on 100 individuals, shows a high degree of skeletal robustness and of occupational markers due to heavy work load (inflammation, muscle-skeletal trauma, fractures, osteoarthritis, enthesopathies). The recording of oral pathologies and aspecific stresses, in order to obtain information about the health status of the population, shows a very high frequency of caries mainly related to poor dental hygiene, and of enamel hypoplasia.

Differences and similarities between arrhythmogenic right ventricular cardiomyopathy and athlete's heart adaptations.

BACKGROUND: Regular intensive physical activity is associated with non-pathological changes in cardiac morphology. Differential diagnosis with arrhythmogenic right ventricular cardiomyopathy (ARVC) constitutes a frequent problem, especially in athletes showing ventricular arrhythmias with left bundle branch block morphology. AIM OF THE STUDY: To assess the different clinical and non-invasive instrumental features of the subjects affected by ARVC and by athletes. METHODS: Three groups of subjects (40 ARVC patients, 40 athletes and 40 controls, mean age 27 (9) years) were examined with family and personal history, physical examination, 12-lead ECG, 24-h ECG, signal-averaged ECG and 2-D and Doppler echocardiography. RESULTS: 12-Lead ECG was abnormal in 62% of ARVC patients versus 7.5% of athletes and 2.5% of controls (p<0.0001). Ventricular arrhythmias and late potentials were present in 70% and 55% of ARVC subjects, respectively (vs 5% of athletes and 7.5% of controls, p<0.0001). Left ventricular parietal wall thickness and left ventricular end-diastolic diameters were significantly higher in athletes. Both athletes and ARVC patients presented a right ventricular (RV) enlargement compared with controls. Moreover, RV outflow tract, measured on parasternal long axis and at the level of aortic root, was significantly larger in ARVC patients (33.6 (4.7) mm vs 29.1 (3.4) mm and 35.6 (6.8) mm vs 30.1 (2.9) mm; p<0.0001), and RV fractional shortening and ejection fraction were significantly lower in ARVC patients compared with athletes (40 (7.9)% vs 44 (10)%; p=0.05 and 52.9 (8)% vs 59.9 (4.5)%; p<0.0001). A thickened moderator band was found to be present in similar percentage in ARVC patients and athletes. CONCLUSIONS: An accurate clinical and instrumental non-invasive evaluation including echocardiography as imaging technique allows to distinguish RV alterations typical of ARVC from those detected in athletes as a consequence of intensive physical activity.

[Evaluation of the motor behavior in rats with cortical ablation]. / Caracterizacion de la conducta motora en ratas con ablacion cortical.

INTRODUCTION: The cortical ablation has been used as an experimental model in order to study the basic mechanisms of functional recovery. However, there is not data concerning to the injury effects on the motor and somatosensorial behavioral manifestations that allow us to categorize such sequels as a hemiplegic model. MATERIALS AND METHODS: We used 35 male Wistar rats (280-300 g) allocated in two groups: control (n = 17) and brain injured by cortical ablation (n = 18). Previously trained, basal recordings of the footprint and motor and somatosensorial assessment were performed in the rats before surgery. The behavioral tests were performed again 6 hours after surgery and the spontaneous ambulatory activity was also evaluated. RESULTS AND CONCLUSIONS: It was observed a decrease in the stride's length and an increase in the stride's angle and in the motor deficit, while the somatosensorial assessment and spontaneous ambulatory activity were not affected. These findings are discussed in function of the motor features of the hemiparetic sequels in humans.