Development of a loop-mediated isothermal amplification assay for specific detection of all known subspecies of Clavibacter michiganensis.

AIMS: Clavibacter michiganensis is an important bacterial plant pathogen that causes vast destruction to agriculturally important crops worldwide. Early detection is critical to evaluate disease progression and to implement efficient control measures to avoid serious epidemics. In this study, we developed a sensitive, specific and robust loop-mediated isothermal amplification (LAMP) assay for detection of all known subspecies of C. michiganensis. METHODS AND RESULTS: Whole genome comparative genomics approach was taken to identify a unique and conserved region within all known subspecies of C. michiganensis. Primer specificity was evaluated in silico and with 64 bacterial strains included in inclusivity and exclusivity panels; no false positives or false negatives were detected. Both the sensitivity and spiked assay of the developed LAMP assay was 1 fg of the pathogen DNA per reaction. A 100% accuracy was observed when tested with infected plant samples. CONCLUSIONS: The developed LAMP assay is simple, sensitive, robust and easy to perform using different detection platforms and chemistries. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed LAMP assay can detect all known subspecies of C. michiganensis. The LAMP process can be performed isothermally at 65°C and results can be visually assessed, which makes this technology a promising tool for monitoring the disease progression and for accurate pathogen detection at point-of-care.

Gas1 is present in germinal niches of developing dentate gyrus and cortex.

Gas1 is a pleiotropic protein that inhibits cell growth when overexpressed in tumors but during development, it acts as a co-receptor for sonic hedgehog to promote the proliferation and survival of various growing organs and systems. This protein has been extensively studied during development in the cerebellum. However, in other structures of the central nervous system, information concerning Gas1 is limited to in situ hybridization studies. We investigate the pattern of Gas1 expression during various developmental stages of the cortex and dentate gyrus of the mouse brain. The levels of Gas1 decrease in the developing brain and the protein is mainly found in progenitor cells during the development of the cortex and dentate gyrus.

Colles' fractures: functional treatment in supination.

PURPOSE OF THE STUDY: Abraham Colles classified and described fractures of the distal epiphyseal radius. He recommended the arm should be immobilized in a cast that extends from the base of the fingers to above the elbow, while holding this joint at ninety degrees of flexion the forearm in pronation and the wrist in slight flexion and ulnar deviation. We identified the brachioradialis muscle as the main culprit in the frequently observed loss of reduction of the fracture. Since the brachioradialis is attached to the distal region of the radius and functions as a flexor of the elbow when the forearm is in pronation, its stimulation easily displaces a reduced fracture, particularly if its geometry suggests axial instability. We concluded that post-reduction stabilization in supination was more desirable than in pronation. MATERIAL AND METHODS: Prospective study of one hundred and fifty-six patients suffering from Colles' fractures who were treated with the functional method. Approximately one-half of the fractures were immobilized in pronation and the other half in supination. The median age of the patients was 49 years. After approximately eleven days of immobilization in an above-the-elbow cast that held the forearm in a relaxed attitude of supination and the wrist in slight flexion and ulnar deviation, a new cast or brace was applied. The appliance permitted flexion of the elbow and slightly limited extension. We utilized modified Lindstom criteria to assess radiological results, according to types of fractures and by groups treated in supination and pronation. RESULTS: In the type I and III (non-displaced) fracture series there appeared to be no significant difference in the functional results between the pronation and supination treated groups. In the type II category, in the supinated fractures, there were 9 excellent, 4 good and no fair or poor results. In the pronated group 9 excellent, 8 good and one fair result. The functional results in type IV fractures treated in supination were excellent in 11 instances, good in 7 and fair in 2. In fractures treated in pronation there were 5 excellent, 10 good and 5 fair results. There were no poor results in either group. 85% of type II fractures and 85% of type IV fractures treated in supination had excellent or good results. In the pronation group, 67% had excellent or good results in type II and 40% in type IV classification. In combining the results for all types of braced Colles' fractures, (I-IV) 93% of the supination group and 87% of the pronation group achieved excellent or good functional results. In analyzing overall results regardless of type of fracture or position of immobilization, 90% of the patients had excellent or good results. CONCLUSION: We treated Colles' fractures in supination and compared the results with those obtained when treated in pronation. The results indicated a lower incidence of re-displacement in the supination group. We developed a forearm brace that permits flexion of the elbow, but prevented pronation of the forearm, and limited extension of the elbow in approximately the last fifteen degrees. It permits minimally limited flexion of the wrist but prevents wrist dorsiflexion. It makes impossible any radial deviation. The place of surgery in the management of Colles' fractures should be limited to those fractures that when treated by non-surgical means are not likely to render satisfactory functional and cosmetic results. There is not at this time a consensus as to when to use the surgical approach. The complication rate from the surgery have not clearly identify superiority of one over the other. Nonetheless, the surgical treatment has a definite place in the armamentarium of the orthopaedic surgeon. In a number of situations, it is the treatment of choice.

[Analysis of the echographic parameters described in the study of carpal tunnel syndrome: A systematic review]. / Análisis de los parámetros ecográficos descritos en el estudio del síndrome del túnel del carpo. Una revisión sistemática.

Carpal tunnel syndrome (CTS) is the most common entrapment mononeuropathy; the diagnosis is established by electrodiagnostic tests with until 34% of false positives/negatives. We present the following systematic review which objective is to analyze the most recent literature related to the ultrasound parameters described to study CTS. We selected studies that evaluated ultrasound parameters in patients with clinical suspicion following the Cochrane manual's recommendations. We include systematic reviews, meta-analyses, case-control studies and diagnostic tests, evaluating retrospective studies and bibliographic reviews with proper methodological quality. Articles published between 2005 and 2019. We included eight articles (two systematic reviews/meta-analyses, two case-control studies, one diagnostic test study, two literature reviews, and one retrospective). The parameters analyzed were cross-sectional area, wrist-forearm index, entry-exit index, thinning range, palmar bowing of the flexor retinaculum, and vascularity/mobility. Current evidence allows us to affirm that ultrasound is useful in screening for CTS.

Bacterial septicemia and herpesvirus infection in Antarctic fur seals (Arctocephalus gazella) stranded in the São Paulo coast, Brazil.

In August 2021, two juvenile male Antarctic fur seals (Arctocephalus gazella) stranded in the southeastern Brazilian coast and were referred to rehabilitation centers. The animals presented increased body temperature, prostration, respiratory distress and despite treatment died. A necropsy following a standardized protocol was performed, and formalin-fixed tissues were processed for microscopic examination. Samples were screened for morbillivirus, herpesvirus, and Brucella spp. by molecular analyses (PCR, RT-PCR). Bacteriological culture was performed in samples collected from the lungs, trachea, and lymph nodes of both cases. The main histopathologic findings were of infectious nature, including multifocal necrotizing and fibrinous mixed interstitial pneumonia, bronchiolitis, and bronchitis, with intralesional myriad bacteria associated with vascular fibrinoid necrosis. Pseudomonas aeruginosa was isolated from tracheal and lung swabs of Case 1, and Klebsiella oxytoca was found in nostril swabs, tracheobronchial lymph nodes, and lung of Case 2. Gammaherpesvirus infection was detected in both cases, and the sequences retrieved were classified into the genus Percavirus. All tested samples were PCR-negative for Brucella spp. and morbillivirus. We hypothesize that the deficient immunological status in association with starvation predisposed the reactivation of herpesvirus and secondary bacterial co-infections. To the authors' knowledge, this is the first molecular detection of herpesvirus in an Antarctic pinniped. These findings reinforce that Otariid gammaherpesvirus circulating in the Southern Hemisphere are likely endemic in the Arctocephalus genus. This report contributes to the current knowledge of health aspects affecting wild pinnipeds, especially in the poorly studied Antarctic species.

Reflections on acromio-clavicular dislocations.

A number of surgical and nonsurgical techniques have been used in the recent past for the treatment of this not uncommon traumatic condition. Thus far, no agreement has been reached regarding the method most likely to consistently render good results. Uppermost in determining the treatment has been the degree of displacement of the clavicle and the prevention of possible cosmetically unacceptable complications. Advances in the surgical care of most fractures and dislocations have lead to the current belief and practice among the majority of orthopaedic surgeons that every effort should be made to correct any deviation from the normal produced by the injury. I submit that skilful neglect and acceptance of acromio-clavicular dislocation is an option worth considering. When the dislocation is accepted, the vast majority of patients do well, functionally and aesthetically. Chronic pain is an extremely rare situation; and the resulting deformity, from the cosmetic point of view, an uncommon problem. If this is the case, what is the problem that reconstructive surgery proposes to address?

Analysis of the effects of different surgical approaches on corporotomy localization in inflatable penile implant surgery performed by expert implant surgeons.

Inflatable penile prostheses may be a solution for patients with erectile dysfunction. To our knowledge, no data exist regarding the effect of different surgical approaches used during implantation on the site of the corporotomy. The main purpose of this multicentre study was to investigate the influence of different surgical approaches on the corporotomy site.Data were collected from six expert implant surgeons. Surgical notes were searched for the incision site, proximal, distal and total corporal length measurement, total cylinder length, length of rear tip extenders, surgery time, type of implant, and reservoir placement. The association between the proximal/distal corporal length and the recorded covariates was examined using a linear mixed model.A total of 1757 patients who underwent virgin prosthesis implantation were included in the analysis. Analysis of proximal/distal measurements was performed on 1709 patients. The proximal/distal ratio had a mean of 0.8 ± 0.3 in penoscrotal incisions (n = 391), 0.7 ± 0.2 in infrapubic incisions (n = 832) and 0.7 ± 0.2 in subcoronal (n = 486) incisions. We observed no significant differences in proximal/distal measurements between the highest-volume surgeons.We could not draw a firm conclusion about the difference in corporotomy site between different surgical approaches, but we found no significant difference between the highest-volume surgeons using different techniques.

Classification and identification of organic binding media in artworks by means of Fourier transform infrared spectroscopy and principal component analysis.

Fourier transform infrared spectroscopy is a powerful analytical technique to study organic materials. However, in Cultural Heritage, since the sample under analysis is always a complicated matrix of several materials, data analysis performed through peak-by-peak comparisons of sample spectra with those of standard compounds is a tedious method that does not always provide good results. To overcome this problem, a chemometric model based on principal component analysis was developed to classify and identify organic binding media in artworks. The model allows the differentiation of five families of binders: drying oils, waxes, proteins, gums, and resins, taking into account the absorption bands in two characteristic spectral windows: C-H stretching and carbonyl band. This new methodology was applied in the characterization of binders in three kinds of artworks: papers of historical, archeological, and artistic value, easel paintings, and polychromed stone-based sculptures.

Estimation of the post-mortem interval of human skeletal remains using Raman spectroscopy and chemometrics.

An important demand exists in the field of forensic analysis to objectively determine the post-mortem interval (PMI) when human skeletal remains are discovered. It is widely known that bones undergo different chemical and physical processes after death, mainly due to their interaction with the environment in which they are found, although it is not known exactly what these processes consist of. Multiple techniques have been used so far to follow up these and other post-mortem changes and thus establish the time elapsed since the individual's death, but they present important drawbacks in terms of reliability and accuracy. The aim of this research was to propose an analytical methodology capable of determining the PMI by using non-destructive Raman spectroscopy measurements of human skeletal remains. The recorded Raman spectra provided valuable and potentially useful information from which a multivariate study was performed by means of orthogonal partial least squares regression (OPLSR) in order to correlate the PMI with the detected spectral modifications. A collection of 53 real human skeletal remains with known PMI (15 years ≤ PMI ≤ 87 years) was analysed and used for building and validating the OPLS model. The PMI of 10 out of 14 validation samples could be determined with an accuracy error of less than 30%, demonstrating the adequate predictive performance of the OPLS model even in spite of the large inter-individual variability it handled. This opens up the possibility of applying the OPLS model in combination with non-destructive techniques to the determination of the PMI of human skeletal remains that have been buried in conditions similar or equal to those of cemetery niches and in a geographic location with a Mediterranean climate, which is an important achievement for forensic medicine and anthropology.

Critically ill COVID-19 patients attended by anesthesiologists in northwestern Spain: a multicenter prospective observational study. / Pacientes críticos COVID-19 atendidos por anestesiólogos en el Noroeste de España: estudio multicéntrico, prospectivo, observacional.

BACKGROUND AND OBJECTIVES: There are limited information on outcome, complications and treatments of critically ill COVID-19 patients requiring admission to an intensive care unit (ICU). The aim of this study is to describe the clinical ICU course, treatments used, complications and outcomes, of critically ill COVID-19 patients admitted in seven ICU in Galicia region during the 2020 March-April pandemic peak. METHODS: Between March 21 and April 19, 2020, we evaluated critically ill COVID-19 patients admitted to the ICU of Anesthesia of seven hospitals in Galicia, northwestern Spain. Outcome, complications, and treatments were monitored until May 6, 2020, the final date of follow-up. RESULTS: A total of 97 critically ill COVID-19 patients were included. During ICU stay, mechanical ventilation became necessary in 80 (82.5%) patients, and tracheostomy in 22 (22.7%) patients. Prone position was used frequently in both intubated (67.5%) and awake (27.8%) patients. Medications consisted of antivirals agents (92.7%), corticosteroids (93.8%), tocilizumab (57.7%), and intermediate or high doses of anticoagulants (83.5%). The most frequent complications were ICU-acquired infection (52.6%), thrombosis events (16.5%), and reintubation (9.3%). After a median follow-up of 42 (34-45) days, 15 patients (15.5%) deceased, 73 patients (75.2%) had been discharged from ICU, and nine patients (9.3%) were still in the ICU. CONCLUSIONS: A high proportion of our critically ill COVID-19 patients required mechanical ventilation, prone positioning, antiviral medication, corticosteroids, and anticoagulants. ICU complications were frequent, mainly infections and thrombotic events. We had a relatively low mortality of 15,5%.

Portable Raman monitoring of modern cleaning and consolidation operations of artworks on mineral supports.

Any restoration performed on cultural heritage artworks must guarantee a low impact on the treated surfaces. Although completely risk-free methods do not exist, the use of tailor-made procedures and the continuous monitoring by portable instrumentation is surely one of the best approaches to conduct a modern restoration process. In this work, a portable Raman monitoring, combined sometimes with spectroscopic techniques providing the elemental composition, is the key analysis technique in the three-step restoration protocol proposed: (a) in situ analysis of the surface to be treated (original composition and degradation products/pollutants) and the cleaning agents used as extractants, (b) the thermodynamic study of the species involved in the treatment in order to design a suitable restoration method and (c) application and monitoring of the treatment. Two cleaning operations based on new technologies were studied and applied to two artworks on mineral supports: a wall painting affected by nitrate impact, and a black crusted stone (chalk) altarpiece. Raman bands of nitrate and gypsum, respectively, decreased after the step-by-step operations in each case, which helped restorers to decide when the treatment was concluded, thus avoiding any further damage to the treated surface of the artworks.

Facial Squamous Cell Carcinoma and Abdominal Peripheral Nerve Sheath Tumour with Rhabdomyoblastic Differentiation in a Rough-toothed Dolphin (Steno bredanensis).

We report the pathological features of a facial squamous cell carcinoma (SCC) and an abdominal peripheral nerve sheath tumour (PNST) with rhabdomyoblastic differentiation in an aged free-ranging rough-toothed dolphin (Steno bredanensis). The animal was found stranded dead in poor body condition. On external examination, there was a 25 × 7 × 3 cm extensively ulcerated area on the right maxillary region of the rostrum, involving the oral mucocutaneous junction with prominent nodular edges, severe soft tissue loss and extensive maxillary and premaxillary bone lysis. On abdominal dissection, a 5 × 4 × 3.5 cm pale tan to red, raised mass expanded the inner aspect of the right transverse abdominis muscle. Microscopically, the aggressive facial lesion was an acantholytic SCC with extensive osteolysis; there was no evidence of metastasis in the tissues examined. The abdominal mass had cytohistomorphological features compatible with a localized PNST, including whorling, Antoni A and Antoni B areas and Verocay bodies intermixed with rhabdomyoblastic components, as suggested by phosphotungstic acid haematoxylin stain. This neoplasm was locally infiltrative, yet no metastases were observed in the tissues examined. No immunohistochemical investigations could be performed due to lack of tissue availability. Total DNA from the formalin-fixed and paraffin wax-embedded SCC was extracted and tested by polymerase chain reaction for herpesvirus and papillomavirus genetic material. There was no amplification for either of these genera. Other pathological findings observed in this animal were related to the 'live-stranding stress response'. The severity and extent of the facial SCC likely related to anorexia and poor body condition and might have played a role in the stranding and death of this dolphin. These two tumour subtypes add to the relatively uncommon reports of neoplasia in cetaceans. Specifically, these appear to be the first neoplasia records for rough-toothed dolphins, including the first documentation of a PNST with features compatible with rhabdomyoblastic differentiation in a marine mammal.

Oxidised proteins and superoxide anion production in the diaphragm of severe COPD patients.

In the diaphragms of chronic obstructive pulmonary disease (COPD) patients, the nature of oxidatively modified proteins and superoxide anion production were explored. Diaphragm specimens were obtained through thoracotomy because of localised lung lesions in COPD patients (16 severe and eight moderate) and 10 control subjects. Lung and respiratory muscle functions were evaluated. Oxidised proteins were identified using immunoblotting and mass spectrometry. Protein and activity levels of the identified proteins were determined using immunoblotting and activity assays. Lucigenin-derived chemiluminescence signals in a luminometer were used to determine superoxide anion levels in muscle compartments (mitochondria, membrane and cytosol) using selective inhibitors. In severe COPD patients compared with controls, respiratory muscle function was impaired; creatine kinase, carbonic anhydrase III, actin and myosin were oxidised; myosin carbonylation levels were increased five-fold; creatine kinase content and activity and myosin protein were reduced; superoxide anion levels were increased in both mitochondria and membrane compartments; and the percentage of superoxide anion inhibition achieved by rotenone was significantly greater. In severe COPD patients, oxidation of diaphragm proteins involved in energy production and contractile performance is likely to partially contribute to the documented respiratory muscle dysfunction. Furthermore, generation of the superoxide anion was increased in the diaphragms of these patients.

Raman spectroscopy after accelerated ageing tests to assess the origin of some decayed products found in real historical bricks affected by urban polluted atmospheres.

Bricks, together with stones and mortars, can be considered as one of the most important building materials that constitute our built heritage. Numerous factors which cause several decaying pathologies in bricks can be listed, but it should be emphasised that the most severe and damaging one is the wet and dry deposition of both combustion and greenhouse gases (CO(2), SO(x) and NO(x) mainly). For instance, after the impact of CO(2) and SO(x), the decayed products promoted in bricks are carbonates and sulphates. Once identified in all these kinds of salts in real samples, it is necessary to make sure that the aggressive atmospheric conditions are sufficient to promote the formation of these salts. Therefore, accelerated exposure test are a good alternative in order to simulate the formation of these decayed compounds and to predict the reactions that promote the decaying mechanism. In this work, brick samples manufactured at different firing temperatures following ancient methods were subjected to humidity/dryness, freeze/thaw, CO(2) and SO(2) (KESTERNICH DIN 50018) accelerated ageing tests followed by a Raman spectroscopy screening in order to verify the formation of sulphate and carbonate salts in bricks on accelerated conditions, simulating the damage caused by a polluted atmosphere throughout many years of exposure.

A surgical technique for the prevention of femoral lysis in cement total hip arthroplasty.

Femoral bone lysis in total hip arthroplasty is thought to be primarily due to polyethylene or metal debris arising from the femoral or acetabular components. The debris appears to gradually seep into the cement/ bone interface, eventually generating the chemical reaction that produces lysis.We experimented with a surgical technique that attempts to construct a proximal bony barrier preventing migration of debris. Following the injection of the acrylic cement and the insertion of the femoral component, but prior to complete polymerization of the cement, bone chips are pressed over the cement, in contact with the viable femoral cortex. The bone chips become rigidly fixed; probably regain viability from the femoral cortex, and seal the proximal femur. In this manner, debris cannot travel into the femoral canal. Although we do not have anatomical evidence that a viable bony seal has formed the absence of lysis and bone/cement radiolucent lines over a period of time ranging from three to fourteen years suggests the permanent presence of a physiological barrier. Attempts to identify the permanency of the bony seal by means of CT scans proved inconclusive. Key words: total hips, lysis, femoral lysis, cortical graft.

The Negro River (Ancash-Peru): A unique case of water pollution, three environmental scenarios and an unresolved issue.

This paper is focused on the hydrogeochemical characterization of the Negro River along its course, as well as in the proposal of a functioning model for the contamination processes in order to establish potential cause-effect relationships between water quality, geology (ARD), mining activities (AMD) and the tectonic framework as transmission vector of acidity, metals and sulphates. The scenario shows a heavily-contaminated river compared to the unaffected regional background. By graphical and statistical treatments of physico-chemical data of Negro River and the unaffected values of regional background and other AMD/ARD representative rivers' it is possible to conclude that Antamina Mine, is not the cause of the Negro River contamination, without the need of isotopic tracers, but just through the inexistent concentrations of Cu, Bi and Mo found in the waters. In the proposed contamination model, climatic factors (glacial retreat) activate geological (ARD) processes. The tectonic scenario (faults) intervenes as a transport medium of the contamination flux from the sulphide oxidation surface in upper altitudes until the spring in lower altitudes. At the end, it is concluded that this contamination comes from the recent glacial retreat in areas near the Cordillera Blanca that has left massive amounts of sulphide materials exposed to weathering conditions, oxidizing naturally (ARD processes) and finally contributing to the contamination of the Negro River through faults. In this case, we would face an ARD process in the strict sense, which is the direct oxidation of sulphides outcropping in the upper part of the mountain with the generation of sulphates, the release of hydrogen ions and the consequent generation of acid and the dissolution of the metals. This ARD process would come from the glacial retreat, which, through the faults, transports contaminated water until the spring.

Analytical diagnosis methodology to evaluate nitrate impact on historical building materials.

Nitrate salts have become of greater importance in the decay of materials from historical buildings due to changes in the environment. This work presents an analytical diagnosis methodology to evaluate the impact of nitrate salts in mortars and bricks, combining noninvasive and microdestructive analytical techniques together with chemometric and thermodynamic data analyses. The impact of nitrate salts cannot be well ascertained if other soluble salts are not taken into account. Therefore, the principal results from this work relate to nitrate salts but some results for other kinds of salts are included. Data from Raman microprobe spectroscopy and micro X-ray fluorescence (micro-XRF) are used to characterise the original composition and a first approximation of the nature of the decay compounds, mainly nitrates. The soluble salts are extracted and the anions and cations are quantified by means of ion chromatography with conductimetric detection for anions/cations and inductively coupled plasma mass spectrometry (ICP/MS) for cations. The values obtained allow two different data treatments to be applied. First, chemometric analysis is carried out to search for correlations among anions and cations. Second, thermodynamic modelling with the RUNSALT program is performed to search for environmental conditions of soluble salt formation. All the results are finally used to diagnose the impact of nitrates.

Functional treatment of closed segmental fractures of the tibia.

PURPOSE OF THE STUDY Segmental tibial fractures are commonly believed to be more difficult to manage, requiring surgical treatment. Our experience with forty-eight segmental tibial fractures suggests that these fractures, if closed and with shortening of an acceptable initial degree and a corrected deformity, may be successfully treated with functional braces. MATERIAL AND METHODS Forty-eight closed segmental fractures of the tibia that had initial shortening .12 mm and angular deformity manually corrected to .7 degrees were stabilized in an above-the-knee cast for a median time of 33 days and a mode of 15 days. They were subsequently stabilized in a functional brace that allowed unencumbered motion of all joints. All other segmental fractures outside the established parameters were managed by other methods. RESULTS All fractures healed at a median time of 15.3 weeks. The final shortening was 4.7 millimeters with a mode of 12 millimeters. The maximum shortening was 14 millimeters. Fractures healed with a medial-lateral (M.L) angular deformities ranging zero to 19 degrees, a median of 5.9 degrees and a mode of 3.4 degrees. CONCLUSION The relatively early introduction of weight bearing and the freedom of motion of all joints that the brace permits seem to result in motion at the fracture site, which in turn enhances osteogenesis. As we have previously documented, the initial shortening that closed tibial fractures experience does not increase with the physiological use of the extremity. The final shortening and angulation observed in most of the fractures should not be considered complications, simply inconsequential deviations from the normal. The same should apply to closed segmental fractures.