From big flow cytometry datasets to smart diagnostic strategies: The EuroFlow approach.

The rise in the analytical speed of mutiparameter flow cytometers made possible by the introduction of digital instruments, has brought up the possibility to manage progressively higher number of parameters simultaneously on significantly greater numbers of individual cells. This has led to an exponential increase in the complexity and volume of flow cytometry data generated about cells present in individual samples evaluated in a single measurement. This increase demands for new developments in flow cytometry data analysis, graphical representation, and visualization and interpretation tools to address the new big data challenges, i.e. processing data files of ≥10-25 parameters per cell in samples with >5-10 million cells (= up to 250 million data points per cell sample) obtained in a few minutes. Here, we present a comprehensive review of some of the tools developed by the EuroFlow consortium for processing flow cytometric big data files in diagnostic laboratories, particularly focused on automated EuroFlow approaches for: i) identification of all cell populations coexisting in a sample (automated gating); ii) smart classification of aberrant cell populations in routine diagnostics; iii) automated reporting; together with iv) new tools developed to visualize n-dimensional data in 2-dimensional plots to support expert-guided automated data analysis. The concept of using reference data bases implemented into software programs, in combination with multivariate statistical analysis pioneered by EuroFlow, provides an innovative, highly efficient and fast approach for diagnostic screening, classification and monitoring of patients with distinct hematological and immune disorders, as well as other diseases.

Automated database-guided expert-supervised orientation for immunophenotypic diagnosis and classification of acute leukemia.

Precise classification of acute leukemia (AL) is crucial for adequate treatment. EuroFlow has previously designed an AL orientation tube (ALOT) to guide towards the relevant classification panel (T-cell acute lymphoblastic leukemia (T-ALL), B-cell precursor (BCP)-ALL and/or acute myeloid leukemia (AML)) and final diagnosis. Now we built a reference database with 656 typical AL samples (145 T-ALL, 377 BCP-ALL, 134 AML), processed and analyzed via standardized protocols. Using principal component analysis (PCA)-based plots and automated classification algorithms for direct comparison of single-cells from individual patients against the database, another 783 cases were subsequently evaluated. Depending on the database-guided results, patients were categorized as: (i) typical T, B or Myeloid without or; (ii) with a transitional component to another lineage; (iii) atypical; or (iv) mixed-lineage. Using this automated algorithm, in 781/783 cases (99.7%) the right panel was selected, and data comparable to the final WHO-diagnosis was already provided in >93% of cases (85% T-ALL, 97% BCP-ALL, 95% AML and 87% mixed-phenotype AL patients), even without data on the full-characterization panels. Our results show that database-guided analysis facilitates standardized interpretation of ALOT results and allows accurate selection of the relevant classification panels, hence providing a solid basis for designing future WHO AL classifications.

Contemporary wars and their contributions to vascular injury management.

INTRODUCTION: Man's inhumanity for man still knows no boundaries, as we continue as a species as a whole to engage in war. According to Kohn's Dictionary of Wars [1], of over 3,700 years of recorded history, there have been a total of 3,010 wars. One is hard pressed to actually find a period of time in which here has not been an active conflict in the globe. The world has experienced two world wars: WWI (1914-1918) and WWII (1939-1945). The total number of military casualties in WWI was over 37 million, while WWII so far, has been the deadliest military conflict in history with over 60 million people killed accounting for slightly over 2.5% of the world's population. MATERIAL AND METHODS: The purpose of this study is to review contemporary wars and their contributions to vascular injury management. It is precisely wartime contributions that have led to the more precise identification and management of these injuries resulting in countless lives and extremities saved. However, surgeons dealing with vascular injuries have faced a tough and arduous road. Their journey was initiated by surgical mavericks which undaunted, pressed on against all odds guided by William Stewart Halsted's classic statement in 1912: "One of the chief fascinations in surgery is the management of wounded vessels." CONCLUSION: Contemporary wars of the XX-XXI centuries gave birth, defined and advanced the field of vascular injury management.

Somnofluoroscopy: cineradiographic observation of obstructive sleep apnea.

Systematic cineradiographic observation of the upper airway with simultaneous polysomnography, termed somnofluoroscopy, was performed for a relatively large sample of obstructive sleep apnea patients. These observations suggest that (a) the site of upper airway occlusion may be oropharyngeal, hypopharyngeal, or both; (b) upper airway dynamics appear to be consistent within a given patient and variable among patients; and (c) a passive mechanism results in occlusion rather than active muscular contraction. Somnofluoroscopy is a relatively simple procedure that may be helpful in the selection of treatment for individual sleep apnea patients.

Partial left ventriculectomy to treat end-stage heart disease.

BACKGROUND: It is reasoned that reducing left ventricular diameter (Laplace's law) in patients with dilated cardiomyopathy, will improve ventricular function. METHODS: Partial left ventriculectomy was performed in 120 patients with end-stage dilated cardiomyopathies of varying causes. Most patients were in New York Heart Association functional class IV. The procedure consisted of removal of a wedge of left ventricular muscle from the apex to the base of the heart. Depending on the distance between the two papillary muscles, the mitral valve apparatus was either preserved, repaired, or replaced with a tissue prosthesis. RESULTS: The 30-day mortality was 22% and the 2-year survival was 55%. Although 10% of surviving patients showed no improvement in New York Heart Association functional class, most of the surviving patients were in either class I (57%) or II (33.3%), and the others were in class III and IV. CONCLUSIONS: Partial left ventriculectomy can be used to treat end-stage dilated cardiomyopathy. Further studies and a longer follow-up period are needed to fully assess the effects of this procedure.

Multimedia and virtual reality techniques in the treatment of male erectile disorders.

The present study is based on the observation that computer-simulated reality applied by virtual reality (VR) methods may offer a new means of treating male erectile disorders. The experimental design was based on the theory of psychological development, supported by multimedia acoustic experience and clinical tests. The method involved the use of virtual reality equipment and specially designed CD-ROM programmes. Excluding 15% drop-outs, the success rate was 82% for male erectile disorders due to psychological factors and 84%, excluding 17% drop-outs, for combined factor disorders. Psychotherapy with VR seems to hasten the healing process and reduce drop-outs, suggesting that this method opens or consolidates new or rarely used brain pathways, facilitating the flow of new mnemonic associations that promote the satisfaction of natural drives.

Environmental assessment of polycyclic aromatic hydrocarbons (PAHs) in surface sediments of the Santander Bay, Northern Spain.

Samples of intertidal surface sediments (0-2 cm) were collected in 17 stations of the Santander Bay, Cantabric Sea, Northern Spain. The concentrations of polycyclic aromatic hydrocarbons (PAHs), 16, were analysed by HPLC and MS detection. Surface sediments show a good linear correlation among the parameters of the experimental organic matter evaluation, where total carbon (TC) and loss on ignition (LOI) are approximately 2.5 and 5 times total organic carbon (TOC). A wide range of TOC from 0.08% to 4.1%, and a broad distribution of the sum of sigma16PAHs, from 0.02 to 344.6 microg/g d.w., which can be correlated by an exponential equation to the TOC, has been identified. A qualitative relationship may be established between the industrial input along the rivers and the concentration of sigma6PAHs in the sediments of the estuaries: Boo estuary (8404-4631 microg/g OC), Solia-San Salvador estuaries (305-113 microg/g OC) and Cubas estuary (31-32 microg/g OC). This work shows a dramatic change in the spatial distribution in the concentration of PAHs of intertidal surface sediments. The left edge of the Bay has the main traffic around the city and the major source of PAHs is from combustion processes and estuarine inputs, leading to medium values of PAHs in the sediments; the right edge of the Bay has much lesser anthropogenic activities leading to lower values of PAHs in sediments. The distribution of individual PAHs in sediments varies widely depending on their structure and molecular weight; the 4-6 ring aromatics predominate in polluted sediments due to their higher persistence. The isomer ratio does not allow any clear identification of the PAHs origin. Environmental evaluation according to Dutch guidelines and consensus sediment quality guidelines based on ecotoxicological data leads to the same conclusion, sediments in the Santander Bay show a very different environmental quality depending on the spatial position from heavily polluted/medium effects to non-polluted/below threshold effects. These results indicate that local sources of PAHs, especially estuary discharges, lead to very different qualities of sediments in coastal zones, where traffic and industrial activities take place.

Birth of normal kids after microinjection of pronuclear embryos in a transgenic goat (Capra hircus) production program in Brazil.

This pilot project was designed to determine if normal kids could be produced after microinjection in pronuclear embryos and subsequent transfer to recipients in a transgenic goat program in Brazil. Twelve donors of the Saanen breed and 17 recipients of an undefined breed were used. The estrus of both donors and recipients was synchronized by a standard progestagen treatment and superovulation obtained by six pFSH injections. Donors in estrus were mated with fertile Saanen bucks. Zygotes were recovered surgically by flushing oviducts. The recovered zygotes with visible pronuclei were microinjected with 500 to 1000 copies of the human G-CSF gene. Two or four embryos were surgically transferred into the oviducts of recipients. One recipient became pregnant and two kids were born. No transgenic goat was identified after PCR analysis. Even though transgenic goats were not obtained, this experiment establishes the basis of a synchronization and superovulation regimen for use in goats raised in Brazil, for the purpose of collecting and manipulating the pronuclear embryos. This project also showed that microinjected one-cell goat embryos can survive to produce live young following surgical transfer.

Environmental assessment of lubricants before and after wire drawing process.

Iron wire drawing processes involve the use of solid lubricants made of powdered raw materials, which lead to industrial wastes after being used. These wastes, based on stearates, have a negative effect on the environment. This study deals with the environmental assessment of some lubricants before and after the wire drawing process in a Spanish factory. The parameters evaluated for this study have been total organic carbon (TOC), mobility of zinc and lead, and ecotoxicity (EC(50)). Results show that wastes have more ecotoxicity than the original lubricants due to the content of metals that lubricants pick up from the wire, as pickling, patenting and galvanising take part in the manufacture. The capture of metallic particles leads to a reduction of TOC and an increase in ecotoxicity.

Pseudopregnancy in Saanen goats (Capra hircus) raised in Northeast Brazil.

The prevalence of pseudopregnancy over 44 months was investigated in 23 Saanen goats raised in Northeast Brazil during continuous oestrous cycling (cyclic group) or after synchronization of oestrus (synchronized group). The goats were monitored by ultrasonography and their plasma progesterone profile. The overall prevalence of pseudopregnancy was 30.4% (7/23). In the cyclic group, 28.6% (4/14) of goats showed pseudopregnancy, while in the synchronized group the prevalence was 33.3% (3/9). There was no significant difference between the groups (p>0.05). The mean (+/- SD) length of pseudopregnancy, as shown by the progesterone profile, was 121.6 +/- 33.5 days, ranging from 70 to 155 days. The study defined the prevalence of pseudopregnancy in Saanen goats raised in Northeast Brazil for the first time. This finding identified a major problem for this breed, as without treatment such animals remain unproductive until the spontaneous resolution of the condition. More research seems desirable to ascertain the prevalence of this condition in other breeds in this region of Brazil.

A VR based therapy for the treatment of impotence and premature ejaculation.

The use of psycho-dynamic psychotherapy integrating virtual reality (VR) dealt with in this study on the treatment of erection dysfunctions and premature ejaculation started several years ago, after having seen the scarce results we obtained using exclusively a psycho-dynamic approach (accompanied by pre-recorded sound and music). Considering the particular way that full-immersion VR involves the subject who experiences it, we hypothesized that better results could be obtained during therapy for these sexual disorders and in particular regarding the nature of erection dysfunction, commonly referred to as impotence "a persistent or recurrent inability to attain, or to maintain until completion of the sexual activity, an adequate erection." The plan for therapy consisted of 12 hour-long sessions over a 25-week period, and the methods involved the use of a VR helmet, joystick and miniature television screens that projected specially-designed CD-ROM programs on psychological development.

Penetrating carotid artery: uncommon complex and lethal injuries.

Carotid arterial injuries are the most difficult and certainly the most immediately life-threatening injuries found in penetrating neck trauma. Their propensity to bleed actively and potentially occludes the airway and makes surgical intervention very challenging. Their potential for causing fatal neurological outcomes demands that trauma surgeons exercise excellent judgment in the approach to their definitive management. The purpose of this article is to review the diagnosis and management of these injuries.

Subclavian vessel injuries: difficult anatomy and difficult territory.

INTRODUCTION: Thoracic and thoracic related vascular injuries represent complex challenges to the trauma surgeon. Subclavian vessel injuries, in particular, are uncommon and highly lethal. Regardless of the mechanism, such injuries can result in significant morbidity and mortality. MATERIALS AND METHODS: Systematic review of the literature, with emphasis on the diagnosis, treatment and outcomes of these injuries, incorporating the authors' experience. CONCLUSIONS: These injuries are associated with significant morbidity and mortality. Patients who survive transport are subject to potentially debilitating injury and possibly death. Management of these injuries varies, depending on hemodynamic stability, mechanism of injury, and associated injuries. Despite significant advancements, mortality due to subclavian vessel injury remains high.

Climacteric complaints, female identity, and sexual dysfunctions.

Forty early menopausal women seeking relief from sexual symptoms within a long-term marital relationship and 40 matched women seeking relief of climacteric complaints completed questionnaires concerning three subject: vasomotor and psychosocial symptoms, sexual dysfunctions, and female identity. Results showed that women with sexual dysfunctions were more likely to suffer from vasomotor and psychosocial complaints and their feminine identity was based mainly on ideals of motherhood and beauty. In addition, sexual desire disorders were present significantly in those women with higher psychosocial symptoms, while sexual arousal disorders were particularly evident in women suffering more vasomotor symptoms.

[The salivary glands of Philodryas patagoniensis Girard, 1857 (Serpentes, Colubridae). A morphological, morphometric and histological study]. / Les glandes salivaires de Philodryas patagoniensis Girard, 1857 (Serpentes Colubridae). Etude morphologique, morphométrique et histochimique.

Morphological, morphometrical and histochemical studies of the cell types in the salivary glands of Philodryas patagoniensis have been performed. It is concluded: 1) the acini of supra, infralabial and premaxillary glands are formed by mucous and mucoserous cells; the Duvernoy's gland by seromucous cells; 2) mucous cells show neutral and sulphated mucosubstances and sialic acid; mucoserous cells show neutral mucosubstance, sialic acid and protein radicals; seromucous cells of Duvernoy's gland show neutral mucosubstance and protein radicals. The acinar area, height of tubule and duct cells, and nuclear volume of acinar, tubule and duct cells were evaluated morphometrically.

Control of adipose tissue lipolysis in ectotherm vertebrates.

Lipolytic activity of fish (Hoplias malabaricus), toad (Bufo paracnemis), and snake (Philodryas patagoniensis) adipose tissue was investigated in vivo and in vitro. Catecholamines or glucagon did not affect the release of free fatty acids (FFA) by incubated fish and toad adipose tissue. Catecholamines also failed to activate snake adipose tissue lipolysis, which even decreased in the presence of epinephrine. However, glucagon stimulated both the lipolytic activity of reptilian tissue in vitro and the mobilization of FFA to plasma when administered to snakes in vivo. The release of FFA from incubated fish, amphibian, and reptilian adipose tissue increased markedly in the presence of cAMP or xanthine derivatives, inhibitors of phosphodiesterase. Forskolin or fluoride, activators of specific components of the adenylate cyclase system, strongly stimulated toad adipose tissue lipolysis. The data suggest that adipocyte triacylglycerol lipase of ectotherm vertebrates is activated by a cAMP-mediated phosphorylation and that the organization of the membrane-bound adenylate cyclase system is similar to that of mammals.