Determining Accessibility of Anterior Cervicothoracic Spine Based on Age and Gender: Radiographic Analysis of Computed Tomography Scans.

Objective The approachability of the cervicothoracic region anteriorly based on age and gender, and the possibility of anatomic variances in different geographic populations have not been previously investigated. The aim of the present work was to perform a radiographic analysis of Brazilian patients to assess anterior approachability of the cervicothoracic junction based on age and gender. Methods Retrospective radiographic analysis of 300 computed tomography scans. Patients were separated based on age and gender. The radiographic parameters studied were: horizontal level above the sternum (HLS), vertebral body angle (VBA), intervertebral disc line (IDL), and intervertebral disc line angulation (IDLA). Results The most frequent HLS and IDL were T2 (34.3%) and C7-T1 (46%) respectively. Vertebral body angleand IDLA had average values of 18 ± 8.94 and 19 ± 7.9 degrees, respectively. Males had higher values in both IDLA ( p = 0.003) and VBA ( p = 0.02). Older groups had higher values in both IDLA ( p = 0.01) and VBA ( p = 0.001). No differences were observed in HLS between gender ( p = 0.3) or age groups ( p = 0.79). No differences were seen in IDL between gender groups ( p = 0.3); however, the older group had a more caudal level than the younger groups ( p = 0.12). Conclusions Compared to other populations, our sample had a more cephalad IDL and HLS. Vertebral body angle and IDLA were higher in males and higher angles for VBA and IDLA were shown for older groups. Intervertebral disc line was more caudal with aging.

Determining Accessibility of Anterior Cervicothoracic Spine Based on Age and Gender: Radiographic Analysis of Computed Tomography Scans / Determinação da capacidade de acesso anterior à coluna cervicotorácica conforme idade e gênero: Análise radiográfica de imagens de tomografia computadorizada

Abstract Objective The approachability of the cervicothoracic region anteriorly based on age and gender, and the possibility of anatomic variances in different geographic populations have not been previously investigated. The aim of the present work was to perform a radiographic analysis of Brazilian patients to assess anterior approachability of the cervicothoracic junction based on age and gender. Methods Retrospective radiographic analysis of 300 computed tomography scans. Patients were separated based on age and gender. The radiographic parameters studied were: horizontal level above the sternum (HLS), vertebral body angle (VBA), intervertebral disc line (IDL), and intervertebral disc line angulation (IDLA). Results The most frequent HLS and IDL were T2 (34.3%) and C7-T1 (46%) respectively. Vertebral body angleand IDLA had average values of 18 ± 8.94 and 19 ± 7.9 degrees, respectively. Males had higher values in both IDLA (p= 0.003) and VBA (p= 0.02). Older groups had higher values in both IDLA (p= 0.01) and VBA (p= 0.001). No differences were observed in HLS between gender (p= 0.3) or age groups (p= 0.79). No differences were seen in IDL between gender groups (p= 0.3); however, the older group had a more caudal level than the younger groups (p= 0.12). ConclusionsCompared to other populations, our sample had a more cephalad IDL and HLS. Vertebral body angle and IDLA were higher in males and higher angles for VBA and IDLA were shown for older groups. Intervertebral disc line was more caudal with aging.

Resumo Objetivo A capacidade de acesso anterior à região cervicotorácica com base na idade e gênero do paciente e a possibilidade de variações anatômicas em diferentes populações geográficas ainda não foram investigadas. O objetivo deste trabalho foi realizar uma análise radiográfica de pacientes brasileiros para avaliar a acessibilidade anterior da junção cervicotorácica conforme idade e gênero. Métodos Análise radiográfica retrospectiva de 300 tomografias computadorizadas. Os pacientes foram separados por idade e gênero. Os parâmetros radiográficos estudados foram: nível horizontal acima do esterno (HLS, na sigla em inglês), angulação do corpo vertebral (VBA, na sigla em inglês), linha do disco intervertebral (IDL, na sigla em inglês) e angulação da linha do disco intervertebral (IDLA, na sigla em inglês). Resultados Os HLS e IDL mais frequentes foram T2 (34,3%) e C7-T1 (46%), respectivamente. Os valores médios de VBA e IDLA foram de 18 ± 8,94 e 19 ± 7,9 graus, respectivamente. Os homens apresentaram valores maiores de IDLA (p= 0,003) e VBA (p= 0,02). Os grupos de maior idade apresentaram valores maiores de IDLA (p= 0,01) e VBA (p= 0,001). Não houve diferenças de HLS entre os gêneros masculino e feminino (p= 0,3) ou faixas etárias (p= 0,79). Não foram observadas diferenças na IDL entre os gêneros masculino e feminino (p= 0,3); entretanto, o grupo mais velho apresentou nível mais caudal do que os grupos mais jovens (p= 0,12). ConclusõesEm comparação a outras populações, nossa amostra apresentou IDL e HLS mais cefálicos. AVBA e a IDLA foram maiores no gênero masculino, enquanto VBA e IDLA foram maiores em grupos mais velhos. A IDL era mais caudal em pacientes idosos.

An invasive mussel is in trouble: How do glyphosate, 2,4-D and its mixture affect Limnoperna fortunei's survival?

Contamination and biological invasions are important factors that alter the functioning of freshwater systems. We carried out two experiments involving daily measurements of dissolved oxygen (DO) and Limnoperna fortunei mortality: 1) the impact of DO on mussel survival mediated by glyphosate, 2,4-D and their mixture was analysed in a 22-day indoor experiment (IE) under controlled conditions using microcosms with L. fortunei, with and without air supply; and 2) the effect of each herbicide and their mixture on mussel accumulated mortality was compared in a 18-day outdoor experiment (OE) using mesocosms without air supply, with and without L. fortunei. Results showed that glyphosate, alone or mixed affected L. fortunei survival both directly and indirectly. In IE we observed direct toxicity of glyphosate in treatments with air supply, with accumulated mortality of 20.0% for glyphosate and 10.0% for the mixture. In OE, L. fortunei deepened the changes in the patterns of DO fluctuations driven by the herbicides, which led to hypoxia in the system. The accumulated mortality was 46.7, 8.6 and 48.2% for glyphosate, 2,4-D and the mixture, respectively. This study contributes to the understanding of the mechanisms that control the invasion of L. fortunei in freshwater systems influenced by agrochemicals.

Invasive species versus pollutants: Potential of Limnoperna fortunei to degrade glyphosate-based commercial formulations.

The intensive use of glyphosate in industrial agriculture may lead to freshwater contamination, encouraging studies of its toxic effect on non-target aquatic organisms. Glyphosate-based commercial formulations contain adjuvants, making them even more toxic than the active ingredient (a.i.) itself. The golden mussel Limnoperna fortunei is a freshwater invasive species which has been found to increase glyphosate dissipation in water and to accelerate eutrophication. The aim of this study is to evaluate the capability of L. fortunei to reduce the concentration of glyphosate in two commercial formulations, Roundup Max® and Glifosato Atanor®. Results were compared with the decay of the a.i. alone and in presence of mussels. Evasive response and toxicity tests were performed in a first set of trials to analyze the response of L. fortunei exposed to Roundup Max® and Glifosato Atanor®. Subsequently, we conducted a 21-day degradation experiment in 2.6-L microcosms applying the following treatments: 6 mg L-1 of technical-grade glyphosate (G), Glifosato Atanor® (A), Roundup Max® (R), 20 mussels in dechlorinated tap water (M), and the combination of mussels and herbicide either in the technical-grade (MG) or formulated form (MA and MR) (all by triplicate). Samples were collected at days 0, 1, 7, 14 and 21. No significant differences in glyphosate decay were found between treatments with mussels (MG: 2.03 ± 0.40 mg L-1; MA: 1.60 ± 0.32 mg L-1; MR: 1.81 ± 0.21 mg L-1), between glyphosate as a.i. and the commercial formulations, and between the commercial formulations, suggesting that the adjuvants did not affect the degrading potential of L. fortunei. In addition to the acceleration of glyphosate dissipation in water, there was an increase in the concentration of dissolved nutrients in water (N-NH4+ and P-PO43-) even higher than that caused by the filtering activity of the mussels, probably resulting from stress or from the degradation of glyphosate and adjuvants. We believe that a larger bioavailability of these nutrients due to glyphosate metabolization mediated by mussels would accelerate eutrophication processes in natural water bodies. The approach used here, where L. fortunei was exposed to two commercial formulations actually used in agricultural practices, sheds light on the potential impact of glyphosate decay on water bodies invaded by this species.

Reliable skeletal maturity assessment for an AIS patient cohort: external validation of the proximal humerus ossification system (PHOS) and relevant learning methodology.

STUDY DESIGN: Validation of classification system. OBJECTIVES: To externally validate the Proximal Humerus Ossification System (PHOS) as a reliable skeletal maturity scoring system and to assess the learning curve associated with teaching the procedure to individuals of varying levels of experience. BACKGROUND: Assessment of skeletal maturity is essential for treatment decisions in Adolescent Idiopathic Scoliosis (AIS). PHOS is a five-stage system that uses the proximal humeral physis in assessing skeletal maturity and has been shown to reliably grade skeletal age leading up to and beyond peak growth age (PGA). This system is advantageous in the AIS patient, as it is often captured in standard scoliosis films. METHODS: A medical student, an orthopedic surgery resident (PGY-2), spine fellow, and experienced scoliosis surgeon in his 25th year in practice were given a three-slide PHOS learning module. Each participant rated 100 X-rays on two separate occasions, separated by 1 week. Intra- and inter-observer reliability, as well as cross-institutional reliability, were calculated using intraclass correlation coefficients (ICC) with 95% confidence intervals [CI95]. RESULTS: Average intra-observer reliability ICC between scoring sessions was 0.94 [0.92, 0.96] and inter-observer reliability by level of training were 0.94 [0.91, 0.96], 0.93 [0.9, 0.95], 0.94 [0.91, 0.96], 0.96 [0.94, 0.97] for the medical student, PGY-2, fellow, and attending, respectively. Reliability across institutions was 0.99 [0.98, 0.99]. Combined rating observations (n = 400) showed 82% exact matches, as well as 17% and 1% mismatches by 1 and 2 stages, respectively. Similar to the PHOS developers, we found PHOS stage 3 to occur immediately after PGA. CONCLUSION: PHOS is easily learned and employed by raters with varying levels of training. It comprises a five-stage system to reliably measure bone age leading up to PGA and thereafter. This new system relies on visualization of the proximal humerus, which is readily available on standard scoliosis X-rays. LEVEL OF EVIDENCE: Level III.

Impact of interaction between Limnoperna fortunei and Roundup Max® on freshwater phytoplankton: An in situ approach in Salto Grande reservoir (Argentina).

The joint impact of the glyphosate-based commercial formulation Roundup Max® and the invasive mussel Limnoperna fortunei on phytoplankton and water quality was assessed in Salto Grande reservoir, a scenario were both stressors coexist. We performed an in situ mesocosm approach, through a 7-day experiment using 400-L enclosures. The following treatments were applied by triplicate: addition of 250 mussels (M); addition of 5 mg L-1 of active ingredient (a.i.) in Roundup Max® (R); addition of 250 mussels and 5 mg L-1 of a.i. in Roundup Max® (MR), and controls, without any addition (C). R showed higher total phosphorus (TP) and ammonium nitrogen (NNH4+) concentrations due to the herbicide input, and a significant increase in algal abundance, biovolume and chlorophyll a levels (Chl-a). In M mussels grazed on phytoplankton, which resulted in subsequent phosphates (SRP) release. A decrease in species diversity was observed in R and M with respect to C. In MR, there were higher TP and NNH4+ concentrations, a decrease in biovolume, an antagonistic effect on Chl-a and a synergistic effect on phytoplankton abundance. Species diversity and evenness showed a significant decrease due to the explosive growth of a small and opportunistic Chlorophyta, Spermatozopsis exsultans. The dominance of this species may be due to negative selectivity for S. exsultans and/or release of potential competitors by L. fortunei, and to the input of nutrients by Roundup Max® and/or removal of competitors by its toxicity.

Biochemical responses of the golden mussel Limnoperna fortunei under dietary glyphosate exposure.

The aim of this study was to analyze the biochemical alterations in the golden mussel Limnoperna fortunei under dietary glyphosate exposure. Mussels were fed during 4 weeks with the green algae Scenedesmus vacuolatus previously exposed to a commercial formulation of glyphosate (6 mg L-1 active principle) with the addition of alkyl aryl polyglycol ether surfactant. After 1, 7, 14, 21 and 28 days of dietary exposure, glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), acetylcholinesterase (AChE), carboxylesterases (CES) and alkaline phosphatase (ALP) activities, glutathione (GSH) content and damage to lipids and proteins levels were analyzed. A significant increase (72%) in the GST activity and a significant decrease (26%) in the CES activity in the mussels fed on glyphosate exposed algae for 28 days were observed. The ALP activity was significantly increased at 21 and 28 days of dietary exposure (48% and 72%, respectively). GSH content and CAT, SOD and AchE activities did not show any differences between the exposed and non exposed bivalves. No oxidative damage to lipids and proteins, measured as TBARS and carbonyl content respectively, was observed in response to glyphosate dietary exposure. The decrease in the CES activity and the increases in GST and ALP activities observed in L. fortunei indicate that dietary exposure to glyphosate provokes metabolic alterations, related with detoxification mechanisms.

Effect of glyphosate acid on biochemical markers of periphyton exposed in outdoor mesocosms in the presence and absence of the mussel Limnoperna fortunei.

Glyphosate is currently the most widely used herbicide in agricultural production. It generally enters aquatic ecosystems through surface water runoff and aerial drift. We evaluated the effect of glyphosate acid on biochemical parameters of periphyton exposed to concentrations of 1, 3, and 6 mg/L in outdoor mesocosms in the presence and absence of the mussel Limnoperna fortunei. Periphyton ash-free dry weight, chlorophyll a content, carotene/chlorophyll a ratio, lipid peroxidation levels, and superoxide dismutase and catalase activities were determined at days 0, 1, 7, 14, and 26 of the experimental period. Ash-free dry weight was similar between control and glyphosate-treated periphyton in the absence of L. fortunei. The latter had significantly lower carotene to chlorophyll a ratios and enzyme activities, and higher lipid peroxidation levels and chlorophyll a content than the former. These results show an adverse effect of glyphosate on the metabolism of periphyton community organisms, possibly inducing oxidative stress. On the contrary, no differences were observed in any of these variables between control and glyphosate-treated periphyton in the presence of L. fortunei. Mussels probably attenuated the herbicide effects by contributing to glyphosate dissipation. The results also demonstrate that biochemical markers provide useful information that may warn of herbicide impact on periphyton communities. Environ Toxicol Chem 2017;36:1775-1784. © 2016 SETAC.

Impact of multiple anthropogenic stressors on freshwater: how do glyphosate and the invasive mussel Limnoperna fortunei affect microbial communities and water quality?

The study of the joint effect of multiple anthropogenic stressors is important because the emerging consequences are often unpredictable on the basis of knowledge of single effects. We explored the joint impact of glyphosate and the invasive golden mussel Limnoperna fortunei on freshwater phytoplankton, bacterioplankton and periphyton, and on the physical and chemical properties of the water. We manipulated both stressors simultaneously in a 25-day experiment using outdoor mesocosms; we assayed technical-grade glyphosate acid at four concentrations: 0, 1, 3 and 6 mg gly L(−1) under scenarios with and without mussels. The addition of the glyphosate significantly increased total phosphorus according to the concentration used; the high clearance rate of L. fortunei significantly decreased phytoplanktonic abundance leading to low values of turbidity. The mussel significantly stimulated the development of filamentous green algae (metaphyton). Interestingly, the combined effect revealed that L. fortunei accelerated the dissipation of glyphosate, which showed a 4-fold decrease in its half-life; this promoted the rapid bioavailability of glyphosate-derived phosphorus in the water. The interaction had a synergistic effect on soluble reactive phosphorus concentrations and was directly dependent on the concentration of glyphosate. A synergistic effect was also observed on bacterioplankton, water turbidity and metaphyton, thus inducing enhanced and rapid eutrophication. The ability of mussels to reduce glyphosate in water may be valued as positive, but our results allow us to predict that the invasion of Limnoperna fortunei in natural freshwater systems contaminated by glyphosate will accelerate the negative impact of the herbicide associated with eutrophication.

Impact of the invasive mussel Limnoperna fortunei on glyphosate concentration in water.

The use of glyphosate has increased dramatically during the past years around the world. Microbial communities are altered when glyphosate reaches water bodies. The freshwater golden mussel Limnoperna fortunei is an invasive species that has rapidly dispersed since it was introduced in Argentina two decades ago. Mussels alter aquatic conditions through their filtrating activity by increasing water clarity and nutrient recycling. We aim to evaluate the potential capacity of the golden mussel to reduce glyphosate concentration in water, in laboratory conditions. Firstly, the evasive response of mussels to glyphosate (10, 20, and 40 mg l⁻¹) was evaluated and a toxicity test was carried out for these concentrations. A three-week experiment was then performed to assess glyphosate variation under mussel presence for two mussel sizes. Finally, mussels' role on glyphosate concentration was evaluated considering different mussel parts (living organisms and empty shells) through another three-week experiment. Laboratory experiments were performed in triplicate using 2-l microcosms. An initial glyphosate concentration between 16 and 19 mg l⁻¹ was used, and when mussels or valvae were added, 20 organisms per aquaria were used. Samples were obtained at days 0, 1, 2, 4, 8, 14, and 21. Glyphosate decreased by 40% under large mussel presence in both experiments, and was reduced by 25% in empty shell treatments. We believe that part of the herbicide that disappears from the water column is adsorbed in valvae surface, while another proportion is being mineralized by microbial communities in shells' biofilm. The mechanisms by which living mussels increase glyphosate dissipation would be degradation, possibly mediated by bacteria associated to mussel's metabolism. Glyphosate half-life depended on mussel and valvae presence and varied with mussel size. L. fortunei presence (either alive or as empty valvae) alters glyphosate concentration in water. We provide preliminary observations from laboratory experiments, with strong potential ecological consequences, about two stressors that could be acting jointly on the environment.

A fibronectin scaffold approach to bispecific inhibitors of epidermal growth factor receptor and insulin-like growth factor-I receptor.

Engineered domains of human fibronectin (Adnectins™) were used to generate a bispecific Adnectin targeting epidermal growth factor receptor (EGFR) and insulin-like growth factor-I receptor (IGF-IR), two transmembrane receptors that mediate proliferative and survival cell signaling in cancer. Single-domain Adnectins that specifically bind EGFR or IGF-IR were generated using mRNA display with a library containing as many as 10 ( 13) Adnectin variants. mRNA display was also used to optimize lead Adnectin affinities, resulting in clones that inhibited EGFR phosphorylation at 7 to 38 nM compared to 2.6 µM for the parental clone. Individual, optimized, Adnectins specific for blocking either EGFR or IGF-IR signaling were engineered into a single protein (EI-Tandem Adnectin). The EI-Tandems inhibited phosphorylation of EGFR and IGF-IR, induced receptor degradation, and inhibited down-stream cell signaling and proliferation of human cancer cell lines (A431, H292, BxPC3 and RH41) with IC 50 values ranging from 0.1 to 113 nM. Although Adnectins bound to EGFR at a site distinct from those of anti-EGFR antibodies cetuximab, panitumumab and nimotuzumab, like the antibodies, the anti-EGFR Adnectins blocked the binding of EGF to EGFR. PEGylated EI-Tandem inhibited the growth of both EGFR and IGF-IR driven human tumor xenografts, induced degradation of EGFR, and reduced EGFR phosphorylation in tumors. These results demonstrate efficient engineering of bispecific Adnectins with high potency and desired specificity. The bispecificity may improve biological activity compared to monospecific biologics as tumor growth is driven by multiple growth factors. Our results illustrate a technological advancement for constructing multi-specific biologics in cancer therapy.