The drop plate method as an alternative for Azospirillum spp viable cell enumeration within the consensus protocol of the REDCAI network

Abstract Quality evaluation of commercial inoculants is essential to warrant an adequate cropresponse to inoculation within a biosecurity framework. In this sense, this work is aimed at standardizing and validating the drop plate method for the enumeration of Azospirillum viable cellsas an alternative to the spread plate technique, which is currently proposed in the consensusprotocol of the REDCAI network. Between 14 and 25 private and public laboratories partici-pated in three independent trials. We obtained consistent and robust results that allowed toconfirm that both techniques are equivalent, concluding that the drop plate method is an alternative enumeration technique that is adequate to be included in the abovementioned consensusprotocol.

Resumen La evaluación de la calidad de los inoculantes comerciales es fundamental para garantizar una adecuada respuesta de los cultivos a la inoculación dentro de un marco de bioseguridad. En este sentido, el objetivo de este trabajo fue la estandarización y validación de la técnica de la microgota para la cuantificación de Azospirillum como metodología alternativa a la técnica de siembra en superficie, propuesta actualmente en el protocolo consenso de la Red de Calidad de Inoculantes, REDCAI. Entre 14 y 25 laboratorios, tanto privados como públicos, participaron de tres ensayos independientes. A partir de ellos se obtuvieron resultados reproducibles y robustos que permiten confirmar que ambas técnicas son equivalentes y concluir que la técnica de recuento por la microgota es una alternativa adecuada para ser incluida dentro del mencionado protocolo consenso.

The drop plate method as an alternative for Azospirillum spp. viable cell enumeration within the consensus protocol of the REDCAI network.

Quality evaluation of commercial inoculants is essential to warrant an adequate crop response to inoculation within a biosecurity framework. In this sense, this work is aimed at standardizing and validating the drop plate method for the enumeration of Azospirillum viable cells as an alternative to the spread plate technique, which is currently proposed in the consensus protocol of the REDCAI network. Between 14 and 25 private and public laboratories participated in three independent trials. We obtained consistent and robust results that allowed to confirm that both techniques are equivalent, concluding that the drop plate method is an alternative enumeration technique that is adequate to be included in the abovementioned consensus protocol.

Experiencias de vida y soporte percibido por las enfermeras de las unidades de hemodiálisis hospitalaria durante la pandemia de COVID-19 en España / Life experiences and support perceived by nurses in hospital haemodialysis units during the COVID-19 pandemic in Spain

OBJETIVO: Profundizar en el conocimiento sobre las experiencias de vida y el soporte percibido por las enfermeras/os que atendieron a pacientes con enfermedad de COVID-19 en tratamiento con hemodiálisis hospitalaria durante los meses de mayor prevalencia de la pandemia en España. MATERIAL Y MÉTODO: Estudio cualitativo fenomenológico. El grupo participante fue de diez enfermeras/o de hospitales públicos de España que habían dializado a pacientes con COVID-19. La recolección de los datos se realizó mediante entrevistas semiestructuradas a través del programa Skype©, hasta conseguir la saturación de las unidades de significado. El análisis se hizo mediante el método de Colaizzi-7 pasos. RESULTADOS: Del análisis de los discursos emergieron cuatro dimensiones asociadas a diferentes subcategorías: desconocimiento sobre la enfermedad, sufrimiento del paciente, soporte percibido y capacidad de afrontamiento de los profesionales. CONCLUSIONES: En las enfermeras han coexistido emociones positivas y negativas. En la etapa álgida del brote epidémico las vivencias de las enfermeras de hemodiálisis fueron provocadas por el desconocimiento, por no saber actuar adecuadamente y por el miedo al contagio, agravado por la falta de medios de protección. Las vivencias positivas fueron el crecimiento personal, el apoyo del equipo y de la familia. También cabe destacar el control racional de la situación, debido a que en las unidades de diálisis hay gran experiencia en el control de la trasmisión de enfermedades infecciosas

AIM: To deepen the knowledge about life experiences and support perceived by nurses who attended in-hospital patients with COVID-19 disease on haemodialysis during the months with the highest prevalence of the pandemic in Spain. MATERIAL AND METHOD: Qualitative phenomenological study. The participating group was ten nurses from public hospitals in Spain who had dialyzed patients with COVID-19. Data collection was performed using semi-structured interviews through Skype© program, until the units of meaning were saturated. The analysis was done using the Colaizzi's seven-step method. RESULTS: Four dimensions emerged from the speech analysis associated with different subcategories: ignorance about the disease, patient suffering, perceived support and professionals' ability to cope. CONCLUSIONS: Positive and negative emotions have coexisted in nurses. In the peak stage of the epidemic outbreak, the experiences of hemodialysis nurses were caused by ignorance, not knowing how to act appropriately, and fear of infection, aggravated by the lack of means of protection. The positive experiences were personal growth, the support of the team and the family. It is also worth noting the rational control of the situation, because in the dialysis units there is great experience in controlling the transmission of infectious diseases

Evaluation of an eye-pointer interaction device for human-computer interaction.

Advances in eye-tracking technology have led to better human-computer interaction, and involve controlling a computer without any kind of physical contact. This research describes the transformation of a commercial eye-tracker for use as an alternative peripheral device in human-computer interactions, implementing a pointer that only needs the eye movements of a user facing a computer screen, thus replacing the need to control the software by hand movements. The experiment was performed with 30 test individuals who used the prototype with a set of educational videogames. The results show that, although most of the test subjects would prefer a mouse to control the pointer, the prototype tested has an empirical precision similar to that of the mouse, either when trying to control its movements or when attempting to click on a point of the screen.

Osteochondral autograft and hemiarthroplasty for bilateral locked posterior dislocation of the shoulder.

Posterior shoulder dislocations are rare and account for less than 4% of all shoulder dislocations, with only 6.6% of these proving to be bilateral. More importantly, the majority of posterior shoulder dislocations continue to represent a diagnostic trap for the unwary physician who first sees patients with this condition. This report describes the treatment of a locked bilateral posterior dislocation of the shoulder with an associated impression fracture of both articular surfaces of the humeral heads due to an epileptic seizure. The osteochondral autograft obtained from the left humeral head was sutured to the dominant (right) side and the hemiarthroplasty was performed on the contralateral non-dominant shoulder. Posterior bilateral dislocation of the shoulder is still a diagnostic challenge. In the chronic setting, bilateral hemiarthroplasty should be postponed as long as possible since posterior bilateral dislocation is usually experienced by middle-aged active males.

Functional and quality-of-life results of displaced and nondisplaced proximal humeral fractures treated conservatively.

OBJECTIVES: Functional and quality-of-life outcomes of conservatively treated proximal humeral fractures. DESIGN: Prospective study. SETTING: University orthopedic department at a hospital. PATIENTS/PARTICIPANTS: Seventy consecutive patients between the ages of 60 and 85 years. INTERVENTION: Conservative treatment. MAIN OUTCOME MEASUREMENTS: Functional outcome measured according to the Constant score, quality of life assessed using EuroQol-5D, and fracture pattern analyzed with x-ray and computed tomography scan. RESULTS: : All fractures consolidated uneventfully with no loss of reduction in either group. Four-part fractures obtained the worst functional results (33.66) followed by three-part fractures (54.64) and finally two-part fractures (65.88 and 71). Mild pain was expected in three- and four-part fractures, whereas two-part fractures achieved near complete pain relief. Nondisplaced fractures obtained a final Constant score of 73.58 and displaced fractures a score of 59.41 with significant differences in all Constant score items with the exception of external rotation. Although patients older than 75 years scored lower (54.63) than those younger than 75 years (70.83), there was no difference in the quality-of-life perception. CONCLUSION: Conservative treatment of proximal humeral fractures in those patients older than age 75 years provides good pain relief with limited functional outcome. Despite limited functional outcome, this appears to have no effect on the quality-of-life perception in the population studied. Four-part fractures present the worst results and treatment options may need to be discussed with the patient to adjust treatment to patient expectations.

Clinical results after anterior mini-open approach for femoroacetabular impingement in early degenerative stage.

We describe our modified anterior mini-invasive technique and results obtained in our first consecutive 117 cases in 105 patients. To assess the influence of preoperative Tönnis degenerative stage, cases were divided into group A (Tönnis 0, 32 hips), B (Tönnis 1, 61 hips), and C (Tönnis 2, 24 hips). The clinical score Dexeus combined score (DCS) was used preoperatively, 3 months, 6 months, and every year after operation. At 3-month follow-up, impingement test results improved significantly in 30 hips of group A (94%; p<0.001) and in 58 of group B (95%; p<0.001), whereas in group C, improvement was observed in only 14 cases (58%; p>0.05). No statistical difference was observed at 3-year visit in all groups. Merle d'Aubigné-Postel and WOMAC scores improved significantly in group A (p<0.001) and B (p<0.001) after 1 year and remained unchanged at subsequent yearly follow-ups. For group C, clinical outcomes scores did not show any significant improvement overall (p>0.05). We concluded that anterior mini-invasive technique is an effective method to treat femoroacetabular impingement, and results are highly influenced by preoperative degenerative state, especially in stage Tönnis 2. Therefore, it seems to be a reasonable early surgical treatment in symptomatic patients.

Morphology of the scapula relative to the reverse shoulder prosthesis.

PURPOSE: To analyse the morphology of the scapula relative to the reverse shoulder prosthesis. METHODS: Scapulas of 46 women and 27 men aged 16 to 84 (mean, 53) years with proximal humeral fractures (n=52) or recurrent antero-inferior instability (n=21) were assessed using 3-dimensional computed tomography (CT). For comparison, 108 cadaveric scapulas with unknown epidemiology were assessed using a goniometer and a caliper. The length of the glenoid neck, the angle between the glenoid surface and the upper posterior column of the scapula, and the angles between the major craneocaudal glenoid axis and (1) the base of the coracoid process and (2) the upper posterior column of the scapula were assessed. RESULTS: The length of the glenoid neck was classified into short and long. In the respective CT and cadaveric groups, the anterior glenoid neck was short in 42% and 18% of scapulas and long in 58% and 82%, whereas the posterior glenoid neck was short in 34% and 60% of scapulas and long in 66% and 40%. The angle between the glenoid surface and the upper posterior column of the scapula was classified into types I (50-52 degrees) and II (62-64 degrees). In the respective CT and cadaveric groups, 61% and 71% of scapulas were type I and 39% and 29% were type II. All differences between groups were significant. The mean angles between the major craneocaudal glenoid axis and (1) the base of the coracoid process and (2) the upper posterior column of the scapula were 18 and 8 degrees, respectively. CONCLUSION: Because of variations in scapular morphology, individualised adjustment is needed for reverse shoulder prostheses. Three-dimensional CT is valuable in preoperative planning.

Cryopreservation does not alter the ultrastructure of the meniscus.

Fresh frozen menisci have recently been shown to have an altered meniscal ultrastructure. The cause might be a deterioration of its permeability due to collagen net disarray. The purpose of this study was to evaluate the cryopreserved meniscus in terms of ultrastructure and cellularity. Ten fresh human lateral menisci were harvested. Collagen architecture was evaluated with transmission electron microscopy. The Collagen Meniscal Architecture scoring system was used to assess the degree of meniscal disarray. Cell population, was also evaluated. The fibril collagen diameters of those menisci which had been previously cryopreserved showed an average size in the longitudinal section of 12.6 +/- 1.3 nm, whereas it was 13.4 +/- 2.2 nm in the menisci used as controls (n.s.). In the transverse section, the cryopreserved menisci averaged 15.5 +/- 2.4 and 16.7 +/- 3.5 nm in the controls (n.s.). The study group scored 4.8 points +/- 1.7, whereas the control group did so at 4.1 +/- 1.3 (n.s.). The percentage of cell survival after the cryopreservation ranged from 4 to 54. The fibril diameters and degree of disarray showed a similar distribution in both groups. The results suggest that meniscal cryopreservation does not alter the meniscal ultrastructure. Therefore, an allograft stored in that way would not alter its biomechanical properties, although its cellular viability is highly unpredictable.

Assessment of radiolucent lines in cemented shoulder hemi-arthroplasties: study of concordance and reproducibility.

We report on the concordance and reproducibility of the evaluation of radiolucent lines in the humeral component of shoulder arthroplasty. Thirty-two shoulder prostheses were assessed independently, on two occasions, by five observers. The level of inter- and intra-observer agreement was calculated using the kappa statistic. Intra-observer agreement: the overall kappa values ranged from 0 to 0.6, meaning poor, fair and moderate agreement levels. Inter-observer agreement: when the anteroposterior (AP) views were analysed, the values obtained for the bone-cement interface ranged from 0.290 to 0.539, meaning a poor-to-moderate agreement. For the cement-implant interface, the values ranged from 0.064 to 0.684, meaning a poor-to-good agreement. When radiolucent lines of the humeral component were analysed, inter-observer agreement proved to be as low as that obtained when total hip or knee components were analysed. Intra-observer agreement showed better results.

Cadaveric and three-dimensional computed tomography study of the morphology of the scapula with reference to reversed shoulder prosthesis.

PURPOSE: The purpose of this study is to analyze the morphology of the scapula with reference to the glenoid component implantation in reversed shoulder prosthesis, in order to improve primary fixation of the component. METHODS: Seventy-three 3-dimensional computed tomography of the scapula and 108 scapular dry specimens were analyzed to determine the anterior and posterior length of the glenoid neck, the angle between the glenoid surface and the upper posterior column of the scapula and the angle between the major craneo-caudal glenoid axis and the base of the coracoid process and the upper posterior column. RESULTS: The anterior and posterior length of glenoid neck was classified into two groups named "short-neck" and "long-neck" with significant differences between them. The angle between the glenoid surface and the upper posterior column of the scapula was also classified into two different types: type I (mean 50 degrees-52 degrees ) and type II (mean 62.50 degrees-64 degrees ), with significant differences between them (p < 0.001). The angle between the major craneo-caudal glenoid axis and the base of the coracoid process averaged 18,25 degrees while the angle with the upper posterior column of the scapula averaged 8 degrees . CONCLUSION: Scapular morphological variability advices for individual adjustments of glenoid component implantation in reversed total shoulder prosthesis. Three-dimensional computed tomography of the scapula constitutes an important tool when planning reversed prostheses implantation.

The pectoralis major tendon as a reference for restoring humeral length and retroversion with hemiarthroplasty for fracture.

The success of anatomic reattachment of the tuberosities in proximal humeral fractures, treated with hemiarthroplasty, correlates with functional results. The purpose of this study was to determine the value of the upper edge of the pectoralis major insertion (PMI) as a landmark to establish the proper height and version of hemiarthroplasty implanted for proximal humeral fractures. Twenty cadaveric humeri were studied by CT scan to analyze the relationship between the PMI and humeral height and retroversion. The mean distance from the PMI to the tangent to the humeral head was 5.64 cm. The mean distance of the PMI to the posterior fin of the prosthesis was 1.06 cm. The mean angle between the PMI and the posterior fin of the prosthesis was 24.65 degrees. The upper edge of the pectoralis major insertion constitutes a reproducible reference point to restore proper humeral height and retroversion in hemiarthroplasty for proximal humeral fracture.

Freezing causes changes in the meniscus collagen net: a new ultrastructural meniscus disarray scale.

Alterations in meniscal permeability leading to nutritional deficit have been suggested as a cause of shrinkage in meniscal transplantation. The purpose of this study was to ascertain how freezing, one of the most common procedures used to preserve meniscal allografts, alters the collagen's architecture. Twenty-six fresh human external menisci were analyzed with transmission electron microscopy. Thirteen of them were previously frozen at -80 degrees C while the rest were used as controls. A new scale of the collagen meniscal architecture was proposed according to the collagen's periodicity and degree of disruption, loss of banding, degree of collagen packing, fibril size variability and its intrafibrilar oedema. Each meniscus was scored from 0 to 7. Subsequently they were classified in grades ranging from a normal state (grade I; 0-2 points) to severe disarray (grade III; 5-7 points). The fibril collagen diameters of those menisci which had been previously frozen showed an average size in the longitudinal section of 14.26 nm, whereas it was 17.28 nm in the menisci used as controls (p=0.019). In the transverse section, the frozen menisci averaged 13.14 and 16.93 nm in the controls (p=0.003). Samples of the 13 previously frozen menisci were classified as grade III in 61.54% of the cases. In the control group, all the menisci were classified either as grade I or II. The frozen menisci averaged 4.85 points, whereas the control group did so at 2.46 (p<0.001). The fibril diameters in frozen menisci showed a thinner diameter and had a higher degree of disarray. Therefore, the results suggest that the freezing process alters the menisci's collagen net. This could partially explain the pathological changes found in shrunken menisci after transplantation.

Promoter 2 -1025 T/C polymorphism in the RUNX2 gene is associated with femoral neck bmd in Spanish postmenopausal women.

Stimulation of bone formation is a key therapeutic target in osteoporosis. Runx2 is a runt domain transcription factor essential to osteoblast differentiation, bone remodeling, and fracture healing. Runx2 knockout mice exhibit a complete lack of ossification, while overexpression of this gene in transgenic mice results in an osteoporotic phenotype. Thus, RUNX2 is a good candidate for the genetic determination of osteoporosis. In this association study, the effects of the -330 G/T polymorphism in promoter 1 and the -1025 T/C polymorphism (rs7771980) in promoter 2 of RUNX2 were tested in relation to lumbar spine (LS) and femoral neck (FN) bone mineral density (BMD) in a cohort of 821 Spanish postmenopausal women. The minor allele frequencies for the two polymorphisms were 0.15 and 0.07, respectively. The two polymorphisms, located more than 90 kb apart, were not in linkage disequilibrium (D' = 0.27, r (2) = 0.028). In an ANCOVA test adjusting by weight, height, age, and years since menopause, the -330 G/T polymorphism was not associated with any of the phenotypes analyzed, while we found the -1025 T/C polymorphism to be associated with FN BMD (p = 0.001). In particular, individuals carrying the TC genotype had higher mean adjusted FN BMD values than those bearing the TT genotype. Our results highlight the importance of this RUNX2 promoter 2 polymorphism in FN BMD determination.

A comparison of risk between the lateral decubitus and the beach-chair position when establishing an anteroinferior shoulder portal: a cadaveric study.

PURPOSE: The purpose of this study was to assess, using a technique that minimally distorts the normal anatomy, the risk of injury when establishing a 5 o'clock shoulder portal in the lateral decubitus versus beach-chair position. METHODS: The anteroinferior portal was simulated with Kirschner wires (K-w) drilled orthogonally at the 5 o'clock position in 13 fresh frozen human cadaveric shoulders. The neighboring neurovascular structures were identified through an anteroinferior window made in the inferior glenohumeral ligament. Their relations to the K-w and surrounding structures were recorded in both positions. RESULTS: The median distance from the musculocutaneous nerve to the K-w was shorter in the lateral decubitus position than in the beach chair position (13.16 mm v 20.49 mm, P = .011). The cephalic vein was closer to the portal in the beach-chair position than in the lateral decubitus position (median 8.48 mm v 9.93 mm, P = .039). The axillary nerve was closer to the K-w in the lateral decubitus position than in the beach-chair position (median 21.15 mm v 25.54 mm, P = .03). No differences in the distances from the K-w to the subscapular and anterior circumflex arteries were found when comparing both positions. The mean percentage of subscapular muscle height from its superior border to the K-w was 53.03%. CONCLUSIONS: This study showed the risk of injury establishing a transubscapular portal in either position. The musculocutaneous nerve and the cephalic vein are the most prone to injury. In general, the beach-chair position proved to be safer. CLINICAL RELEVANCE: Inserting anchor devices orthogonally would permit stronger fixation but presents the risk of damaging neurovascular structures. This study focused on showing the neurovascular risk of performing full orthogonal insertion. Considering the good results reported with the usual superior-anterior portals, we do not recommend performing a transubscapular portal in routine shoulder arthroscopy.

Micropropagation of photinia employing rhizobacteria to promote root development.

An alternative protocol was developed for in vitro propagation of photinia (Photinia x fraseri Dress), an ornamental shrub, using the plant growth-promoting rhizobacteria (PGPR) Azospirillum brasilense and Azotobacter chroococcum during rhizogenesis. Shoot tips from four-year-old mature plants, cut in spring and summer, were used as initial explants. They were cultured on Murashige-Skoog (MS) medium with Gamborg's vitamins, N(6)-benzyladenine (BA: 11.1 microM) and gibberellic acid (GA(3): 1.3 microM), obtaining 63% of established explants. The highest shoot length (22.9 mm) and multiplication rate (4.3) was achieved by cultivating for four weeks in the same basal medium supplemented with 4.4 microM BA. Both auxin induction and bacterial inoculation were used for rooting. Elongated shoots were treated with two concentrations of indole-3-butyric acid (IBA: 4.9 or 49.2 microM) during 6 days for auxin induction. Then, the shoots were transferred to an auxin-free medium and inoculated with A. brasilense Cd, Sp7 or A. chroococcum (local strain). Bacterial inoculation induced earlier rooting of photinia shoots. A. brasilense Cd with 49.2 microM IBA pulse showed a significant increase (P

The influence of the acromial coverage index in rotator cuff tears.

Several intrinsic and extrinsic factors have been advocated in the pathogenesis of rotator cuff tears, but it is still unclear whether the origin of the tear is related to tendon degeneration itself or induced by several morphologic changes. The purpose of this study is to determine the relationship between the acromial coverage of the humeral head and the presence of a cuff tear. We evaluated 148 shoulders, including 45 that underwent surgical rotator cuff repair (group I), 26 with documented rotator cuff tears treated conservatively (group II), and 77 with no cuff pathology as a control group (group III). The mean acromial coverage index was 0.68 in group I, 0.72 in group II, and 0.59 in group III, giving a highly significant difference (P < .0001) between the control group and both cuff tear groups. Patients with a cuff tear have a significantly higher acromial coverage index than the control group.

Innervation patterns of the inferior glenohumeral ligament: anatomical and biomechanical relevance.

Although the Inferior Glenohumeral Ligament (IGHL) has a well known mechanical and proprioceptive relevance in shoulder stability, the interrelation of the ligament's anatomical disposition/innervation has not actually been described previously. The purpose of the study was to determine the IGHL innervation patterns and relate them to dislocation. Forty-five embalmed and 16 fresh-frozen human cadaveric shoulders were studied. Masson's Trichrome staining detailed the intraligamentous nerve fiber arrangements. The effect on the articular nerves of an anteroinferior dislocation of the shoulder joint and the position of 60 degrees abduction and 45 degrees external rotation was studied dynamically. The axillary nerve provided IGHL innervation in 95.08% of the cases. We saw two distinct innervation patterns originating from the axillary nerve. In Type 1, one or two collaterals diverged later from the main trunk to enter the ligament. Type 2 showed innervation to the ligament provided by the posterior branch for three to four neural branches. In both cases, these branches enter the ligament near the glenoid rim and at the 7 o'clock position (right shoulder). The radial nerve (Type 3 innervation pattern) provided IGHL innervation in 3.28% of the cases. Microscopic analysis revealed wavy intraligamentous neural branches. The articular branches relaxed and separated from the capsule at the apprehension position and stayed intact after dislocation. These results showed a special predisposition to avoid possible denervation and suggested that the neural arch probably remains unaffected after most dislocations. Knowledge of the neural anatomy of the shoulder will clearly help in avoiding its injury in surgical procedures.

Proceso de atención de enfermería a un paciente con una Insuficiencia Renal Crónica descompensada y con Patologia Psicosocial Asociada

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