Implementation of Static Telecytology Applications for Accreditation Purposes According to ISO 15189

Telecytology can be defined as the process of diagnostic cytology performed on digital images. The process of diagnostic cytology performed on digital images can be used for teaching, professional examinations, obtaining expert opinions on severe cases, and routine diagnosis of the entire laboratory workload. Ηowever, little information exists about its possible use for proficiency testing purposes. The existing studies have found a high (90%–95%) concordance between telecytological and glass slide diagnoses, but they refer to a small number of cases. Thus, the purpose of the chapter is to examine the feasibility of developing a proficiency testing scheme for cytology labs wishing to be accredited according to ISO 15189:2012 by using low-cost telemedical applications.

Pathogenesis and methods of diagnosis of chronic kidney disease in cats: retrospective analysis (1981–2007)

The aim: to conduct a retrospective analysis of literature sources on the pathogenesis and methods of diagnosis of chronic kidney disease in cats. Materials and methods. The research was conducted by the method of scientific literature open-source analysis: PubMed, Elsevier, electronic resources of the National Library named after V.I. Vernadsky (1981–2007). Results. Chronic kidney disease is a common reason for cat owners to go to veterinary clinics. The term “chronic kidney disease” has a broader meaning than the more limited and not very specific name – chronic renal failure; it is also used to indicate the preazotemic stage of the disease. Chronic kidney disease is characterized by a gradual deterioration of the clinical condition of animals due to progressive decline in renal function. An idea of the pathogenesis and methods of diagnosis of chronic kidney disease in the period from 1981 to 2007 is presented. Conclusions. According to the results of retrospective analysis of literature sources for the period from 1981 to 2007, the basis was identified aspects of the pathogenesis of chronic kidney disease in domestic cats, which have not lost relevance today. The main link during chronic kidney disease in cats is the development of hyperazotemia and, as a consequence, endogenous intoxication of the body, which develops gradually and leads to the death of the animal. The morphological basis of chronic kidney disease in cats is the development of diffuse nephrosclerosis, which is reflected in the results of clinical, biochemical and instrumental studies. According to biochemical analysis of blood, in cats recorded an increase in urea and creatinine, the results of clinical studies of urine showed a decrease in its relative density, as well as the development of proteinuria, the appearance of erythrocytes and cylinders. According to the results of hematological research, anemic syndrome develops due to decreased erythropoietin synthesis. With age in cats, ultrasound examination of the kidneys reveals a decrease in their volume due to uniform sclerosis of the parenchyma: it is determined by its thinning and increased echogenicity due to the accumulation of connective tissue components, which is a sign of nephrosclerosis. Although kidney biopsy is the most informative method of diagnosing chronic kidney disease, it has many contraindications, which does not allow its use in the routine diagnosis of nephropathy in domestic cats. its thinning and increase in echogenicity due to the accumulation of connective tissue components, which is a sign of nephrosclerosis, is determined. Although kidney biopsy is the most informative method of diagnosing chronic kidney disease, it has many contraindications, which does not allow its use in the routine diagnosis of nephropathy in domestic cats. Its thinning and increase in echogenicity due to the accumulation of connective tissue components, which is a sign of nephrosclerosis, is determined

GC-rich repeat expansions: associated disorders and mechanisms

Noncoding repeat expansions are a well-known cause of genetic disorders mainly affecting the central nervous system. Missed by most standard technologies used in routine diagnosis, pathogenic noncoding repeat expansions have to be searched for using specific techniques such as repeat-primed PCR or specific bioinformatics tools applied to genome data, such as ExpansionHunter. In this review, we focus on GC-rich repeat expansions, which represent at least one third of all noncoding repeat expansions described so far. GC-rich expansions are mainly located in regulatory regions (promoter, 5′ untranslated region, first intron) of genes and can lead to either a toxic gain-of-function mediated by RNA toxicity and/or repeat-associated non-AUG (RAN) translation, or a loss-of-function of the associated gene, depending on their size and their methylation status. We herein review the clinical and molecular characteristics of disorders associated with these difficult-to-detect expansions.

A Comprehensive Analysis of the Genetic Diversity of Plasmodium falciparum Histidine-Rich Protein 2 (PfHRP2) in the Brazilian Amazon

Early diagnosis and treatment are fundamental to the control and elimination of malaria. In many endemic areas, routine diagnosis is primarily performed microscopically, although rapid diagnostic tests (RDTs) provide a useful point-of-care tool. Most of the commercially available RDTs detect histidine-rich protein 2 (HRP2) of Plasmodium falciparum in the blood of infected individuals. Nonetheless, parasite isolates lacking the pfhrp2 gene are relatively frequent in some endemic regions, thereby hampering the diagnosis of malaria using HRP2-based RDTs. To track the efficacy of RDTs in areas of the Brazilian Amazon, we assessed pfhrp2 deletions in 132 P. falciparum samples collected from four malaria-endemic states in Brazil. Our findings show low to moderate levels of pfhrp2 deletion in different regions of the Brazilian Amazon. Overall, during the period covered by this study (2002-2020), we found that 10% of the P. falciparum isolates were characterized by a pfhrp2 deletion. Notably, however, the presence of pfhrp2-negative isolates has not been translated into a reduction in RDT efficacy, which in part may be explained by the presence of polyclonal infections. A further important finding was the discrepancy in the proportion of pfhrp2 deletions detected using two assessed protocols (conventional PCR versus nested PCR), which reinforces the need to perform a carefully planned laboratory workflow to assess gene deletion. This is the first study to perform a comprehensive analysis of PfHRP2 sequence diversity in Brazilian isolates of P. falciparum. We identified 10 PfHRP2 sequence patterns, which were found to be exclusive of each of the assessed regions. Despite the small number of PfHRP2 sequences available from South America, we found that the PfHRP2 sequences identified in Brazil and neighboring French Guiana show similar sequence patterns. Our findings highlight the importance of continuously monitoring the occurrence and spread of parasites with pfrhp2 deletions, while also taking into account the limitations of PCR-based testing methods associated with accuracy and the complexity of infections.

Atypical Pupil Reactions in Brain Dead Patients

Background: During routine diagnosis of brain death, changes in pupil diameter in response to the stimulation of peripheral nerves are sometimes observed. For example, pupillary dilation after diagnosed brain death is described in the literature as the ciliospinal reflex. However, pupil constriction creates diagnostic doubts. Objective: The pupillometric analysis of pupil response to stimulation of the cervicothoracic spinal cord in patients with diagnosed brain death. Methods: Instrumental tests to confirm the arrest of cerebral circulation were performed in 30 adult subjects (mean age 53.5 years, range 26–75 years) with diagnosed brain death. In addition, a pupillometer was used to measure the change in pupil diameter in response to neck flexion. Intervention: Flexion of the neck and measuring the response in change of the pupil with the use of the pupillometer. Results: The change in the pupil was observed in the examined group of patients. Difference in pupil size ≥ 0.2 mm was observed in 14 cases (46%). In five cases (17%), pupil constriction was found (from 0.2 to 0.7 mm). Measurement error was +/− 0.1 mm. Conclusions: Both pupillary constriction and dilatation may occur due to a ciliospinal reflex in patients with brain death. This phenomenon needs further research in order to establish its pathophysiology.

Validation of grading of non-invasive urothelial carcinoma by digital pathology for routine diagnosis

Background Pathological grading of non-invasive urothelial carcinoma has a direct impact upon management. This study evaluates the reproducibility of grading these tumours on glass slides and digital pathology. Methods Forty eight non-invasive urothelial bladder carcinomas were graded by three uropathologists on glass and on a digital platform using the 1973 WHO and 2004 ISUP/WHO systems. Results Consensus grades for glass and digital grading gave Cohen’s kappa scores of 0.78 (2004) and 0.82 (1973). Of 142 decisions made on the key therapeutic borderline of low grade versus high grade urothelial carcinoma (2004) by the three pathologists, 85% were in agreement. For the 1973 grading system, agreement overall was 90%. Conclusions Agreement on grading on glass slide and digital screen assessment is similar or in some cases improved, suggesting at least non-inferiority of DP for grading of non-invasive urothelial carcinoma.

Contribution of lung ultrasound in diagnosis of community-acquired pneumonia in the emergency department: a prospective multicentre study

Lung ultrasound (LUS) can help clinicians make a timely diagnosis of community-acquired pneumonia (CAP).ObjectivesTo assess if LUS can improve diagnosis and antibiotic initiation in emergency department (ED) patients with suspected CAP.DesignA prospective observational study.SettingsFour EDs.ParticipantsThe study included 150 patients older than 18 years with a clinical suspicion of CAP, of which 2 were subsequently excluded (incorrect identification), leaving 148 patients (70 women and 78 men, average age 72±18 years). Exclusion criteria included a life-threatening condition with do-not-resuscitate-order or patient requiring immediate intensive care.InterventionsAfter routine diagnostic procedure (clinical, radiological and laboratory tests), the attending emergency physician established a clinical CAP probability according to a four-level Likert scale (definite, probable, possible and excluded). An LUS was then performed, and another CAP probability was established based on the ultrasound result. An adjudication committee composed of three independent experts established the final CAP probability at hospital discharge.Primary and secondary outcome measuresPrimary objective was to assess concordance rate of CAP diagnostic probabilities between routine diagnosis procedure or LUS and the final probability of the adjudication committee. Secondary objectives were to assess changes in CAP probability induced by LUS, and changes in antibiotic treatment initiation.ResultsOverall, 27% (95% CI 20 to 35) of the routine procedure CAP classifications and 77% (95% CI 71 to 84) of the LUS CAP classifications were concordant with the adjudication committee classifications. Cohen’s kappa coefficients between routine diagnosis procedure and LUS, according to adjudication committee, were 0.07 (95% CI 0.04 to 0.11) and 0.61 (95% CI 0.55 to 0.66), respectively. The modified probabilities for the diagnosis of CAP after LUS resulted in changes in antibiotic prescriptions in 32% (95% CI 25 to 40) of the cases.ConclusionIn our study, LUS was a powerful tool to improve CAP diagnosis in the ED, reducing diagnostic uncertainty from 73% to 14%.Trial registration numberNCT03411824.

Reflected Light Spectrometry and AI-Based Data Analysis for Detection of Rapid Chicken Eggshell Change Caused by Mycoplasma Synoviae

Mycoplasma synoviae (MS) is a pathogen that causes economic losses in the poultry industry. It can be transmitted, amongst others, via the respiratory tract and spread relatively quickly. As such, MS infections are mainly controlled by maintaining MS-free breeder flocks. Routine diagnosis for the detection of MS may be based on serological, culture, and molecular tests. Here, we propose an optical solution where AI-based analysis of spectral data obtained from the light reflected from the eggshells is used to determine whether they originate from healthy or Mycoplasma synoviae-infected hens. The wavelengths proposed for spectral MS detection are limited to those of VIS and NIR DPSS lasers, which are freely accessible on market. The results are satisfactory: for white eggshells, the F-score is over 95% for five different combinations of wavelengths (using eight or nine wavelengths); for brown eggshells, the F-score is above 85%, also for five different combinations of 6–9 wavelengths.

Detection of Wheat Lipid Transfer Protein (Tri a 14) Sensitization: Comparison between Currently Available Diagnostic Tools

<b><i>Background:</i></b> Wheat lipid transfer protein (LTP; Tri a 14) and ω5-gliadin have been described as major allergens in wheat allergy (WA) and relevant in wheat-induced anaphylaxis, frequently associated with cofactors. <b><i>Objective:</i></b> The objective of this study was to compare tools currently available in routine diagnosis to detect Tri a 14 sensitization, its clinical relevance, and cosensitization to ω5-gliadin and other LTPs. <b><i>Methods:</i></b> One hundred eighteen adults sensitized to rTri a 14 by ImmunoCAP^® (cutoff ≥0.1 kU_A/L) identified among 210 LTP allergic patients were included. We evaluated (1) wheat skin prick test (SPT), (2) specific IgE (sIgE) to wheat, rTri a 14, rTri a 19, peach, apple, walnut, hazelnut, and peanut LTPs using ImmunoCAP^® and microarray ImmunoCAP®ISAC (cutoff ≥0.3I SU), and (3) wheat-related symptoms. <b><i>Results:</i></b> Wheat SPT and sIgE were positive in 31% and 85% of subjects, respectively. rTri a 14 by microarray was detected in 25%. Eight percent showed cosensitization to ω5-gliadin. Thirty percent referred symptoms (gastrointestinal [13%], urticaria [11%], and anaphylaxis [8%]). Cofactors (45%) were significantly associated with systemic reactions. <b><i>Conclusion:</i></b> WA due to Tri a 14 is frequently related with systemic reactions and because are frequently related to cofactors, the culprit may not be suspected. Together with the poor performance to identify Tri a 14 sensitization of the current routine diagnostic tools based on the analysis of whole wheat extract, such as wheat SPT or sIgE, there is a high risk that WA may be overlooked. Thus, when WA is suspected, sIgE Tri a 14 assessment is recommended, together with wheat and ω5-gliadin, preferably in the singleplex format, and carefully evaluated considering ≥0.1 kUA/L as a cutoff.

Implementation of Mini-FLOTAC in Routine Diagnosis of Coccidia and Helminth Infections in Domestic and Exotic Birds

Mini-FLOTAC (MF) has recently been proposed for the fecal quantification of gastrointestinal (GI) parasites in birds due to its higher sensitivity and precision in comparison with the McMaster method. The current research aimed to test the use of MF in routine diagnosis of coccidia and helminth infections in several domestic and exotic bird collections in Portugal. Between July 2020 and April 2021, a total of 142 fecal samples from organic layers, peacocks and ratites were collected in four Portuguese bird collections and processed using MF and fecal cultures to identify and calculate GI parasite shedding and prevalence. The McMaster method was also used to compare the shedding levels obtained for both quantitative techniques. MF’s relative sensitivity and specificity were also assessed, using McMaster as the reference technique. The implementation of MF resulted in an average Eimeria spp. shedding higher in peacocks from bird collection 2 (502 OPG), followed by peacocks from collection 1 (107 OPG) and organic layers (24 OPG) and peacocks from collection 3 (9 OPG). Peacocks were also positive for Capillaria spp., Trichostrongylus tenuis and Strongyloides pavonis, whereas ostriches and emus were infected by L. douglassii. The MF protocol for exotic animals and the McMaster method did not differ significantly for each parasitic agent and bird species, and MF achieved relative sensitivities and specificities higher than 70% for Galliform Eimeria spp., peacock helminths and ratites’ L. douglassii infections. Higher L. douglassii EPG values were identified using the MF protocol for exotic species (2 g of feces/38 mL of sucrose solution), followed by McMaster 2/28, MF 5/45 and MF 2/18. The use of MF allowed for obtaining different intestinal parasitic populations in several bird species and locations, and MF 2/38 is globally proposed as the most suitable protocol for bird fecal samples as an alternative to the McMaster method in the diagnosis of avian intestinal parasitic infections.