Clinical Laboratory Analysis Research Articles

Sex Differences in Cardiovascular Markers and BDNF between Cerebral Palsy and Mild Cognitive Impairment

A diverse array of aging clocks, derived from a variety of omics data and clinical biomarkers, have been developed to describe aging and predict age-related disease. As such, these biomarkers are particularly applicable for use in observational studies, basic science and clinical trials focused on tackling biological aging. However, ongoing research suggests significant heterogeneity in aging, with deterioration and disease occurring in different organ systems or functional domains at various rates across individuals. Existing aging clocks only measure heterogeneity in the degree of aging, not in the manner of aging (e.g. different organ systems or functional domains). We hypothesize these unique trajectories exist and that they can be captured using a systems based approach. In our work, using clinical chemistry biomarkers from participants in the Health and Retirement Study (HRS), Framingham Heart study (FHS) and Women’s Health Initiative (WHI) , we modeled unique epigenetic aging trajectories from distinct groups of biological processes (such as Immune function, metabolic function, hepatic function, cardiac function, renal function and more). Interestingly, these biological system specific scores when combined gave an aging clock with superior mortality prediction than any published aging clock. We further validate the system aging scores and aging clock in different clinical studies to show the added advantage of such a measure, such as the fact that people with similar epigenetic age may have very different system scores. Overall, this method introduces the potential for quantitative and multi-dimensional, personalized aging scores that are indicative of an individual’s disease and disorder risk.

Crusted Scabies as a Suitable Disease for Teledermatology: A Study of 2 Cases

Background Teledermatology has been available for several years now, but the COVID-19 pandemic has highlighted its importance, especially in remote communities. Crusted scabies (CS) presents a unique clinical picture that favors telediagnosis. Patients with neurological diseases, as well as homeless, HIV-infected patients and people with impaired immunological function, are at risk. Clusters of CS have been reported in French Guyana, and these were associated with human T-lymphotropic virus infections. CS has also been reported in Aboriginal Australian communities. Objective Teledermatology is especially useful in cases of CS, as it is a disease that affects areas that are in need of medical services. At the same time, CS presents a unique clinical picture. The objective of this presentation is to fuel the clinical suspicion and detection of patients with this debilitating condition. Methods Relatives of patient 1 contacted our clinic for teledermatology appointments. General practitioners from health services sent images of the second patient. Results Case 1 involved an older woman living in a nursing home with Alzheimer disease, which was severe enough to constrain her to bed. We recommended that her relatives (who had sent images) collect skin scrapings in a container. These scrapings were sent to a clinical analysis laboratory where microscopic potassium hydroxide preparation revealed the presence of Sarcoptes mites. Treatment with oral ivermectin and topical permethrin resulted in the complete resolution of the lesions. Case 2 involved a homeless, HIV-positive, 42-year-old male. The images were sent by clinicians from local health services. This patient was also treated with oral ivermectin and permethrin lotion. We recognize that this case would need further diagnostic workup, but it is highly indicative of CS. Conclusions CS is one of the most suited diseases for the practice of teledermatology for widespread, large, hyperkeratotic fissured plaques covered with abundant, silvery scales for which the expression “once seen, never forgotten” is highly applicable. These cases are gratifyingly simple to treat, and patients benefit from rapid clinical improvement. Prompt diagnoses prevent outbreaks of scabies for relatives and medical personnel, since these skin crusts contain large numbers of scabies mites. CS has been increasingly reported but poorly recognized, and it has often been misdiagnosed as psoriasis. Images, such as those shown in this presentation, are unique and are enough to raise strong clinical suspicion. Conflicts of Interest None declared.

569 Advanced heart failure patients: insight clinical characteristics and prognosis

Abstract Aims The prevalence of patients with advanced heart failure (HF) ranges from 1% to 10% of HF population. A recent position statement of Heart Failure Association (HFA) of European Society of Cardiology (ESC), defined advanced HF according four criteria including LV dysfunction, symptoms severity, and HF hospital admission.1 In this study we would like to evaluate clinical and prognostic characteristics of HF patients in advanced stage. Methods and results This is an observational retrospective study enrolling patients with diagnosis of acute heart failure (AHF) de novo or not, admitted to our department from January 2015 to January 2019 within 12 h from emergency department admission. Patients underwent to clinical examination, laboratory analysis (NTproBNP, renal function haemoglobin) and echocardiography. Advanced heart failure patients were defined on the basis of ESC recent criteria.1 Patients were followed for 1 year after hospital discharge for the composite outcome of HF re-hospitalization and cardiovascular death through 1 year. A total of 601 AHF patients were included in this analysis. Median age was 78 (70–83) years, median of left ventricular ejection fraction (LVEF) was 45 (33–55)% and the median of serum levels of NTproBNP was 7851 (3222–17 543) pg/ml. In our sample we found 122 patients who met the criteria of advanced HF. Comparing patients with advanced HF and without we found that advanced HF patients were more frequent affected by not ischaemic cardiomyopathy with respect to patients not advanced (85% vs. 49%; P < 0.001). Moreover, patients not advanced were more frequent de novo HF with respect to advanced ones (54% vs. 34%; P < 0.001). Advanced heart failure patients showed higher values of NTproBNP [12 375 (5346–26 375) vs. 6652 (2552–13 782) pg/ml; P < 0.001] and creatinine [1.20 (0.88–1.99) vs. 1.07 (0.82–1.50) mg/dL; P = 0.002] and lower values of TAPSE [18 (15–20) vs. 16 (13–19) mm; P < 0.001], eGFR [48 (30–66) vs. 52 (38–38) ml/min/m2; P = 0.05], and serum sodium [140 (138–143) vs. 139 (136–141) mEq/l; P = 0.001] with respect to not advanced HF patients. Univariate analysis showed that advanced HF was related to poor prognosis in terms of one year cardiovascular death or HF re-hospitalization [HR: 1.83 (1.32–2.54); P < 0.001] and in terms of in-hospital mortality [HR: 2.53 (1.01–6.53); P = 0.05]. Conclusions Advanced HF patients showed a worse neuro-hormonal and renal pattern compared to not advanced ones. Similarly, these patients experienced a worse right ventricular function and were more prone no adverse events development at 1 year. Further study are mandatory to better manage these patients improving their outcome.

570 Prevalence and prognosis of cardiac cachexia in acute heart failure patients

Abstract Aims Cachexia is characterized by a pathological shift of metabolism towards a catabolic state. The prevalence of cardiac cachexia in heart failure (HF) patients is around 10% and it recognizes a negative prognostic impact. In this study we would like to evaluate prevalence and prognosis of cardiac cachexia in acute heart failure (AHF) patients. Methods and results This is an observational retrospective study enrolling patients with diagnosis of acute heart failure (AHF) de novo or not, admitted to our department from January 2015 to September 2018 within 12 h from emergency department admission. Patients underwent to clinical examination, laboratory analysis and echocardiography. Cardiac cachexia was defined as unintentional weight loss, with or without skeletal muscle wasting, of at least 5% of baseline weight during the previous year. For the diagnosis, three of the following factors are also required: anorexia, fatigue, reduced muscle strength, reduced fat-free mass index, and abnormalities in blood biomarkers (haemoglobin ≤12 g/dl, serum albumin <3.2 g/dl, elevated IL-6, or increased C-reactive protein).1 Patients were followed for 1 year after hospital discharge for the composite outcome of HF re-hospitalization and cardiovascular death through 1 year. A total of 415 AHF patients were included in this analysis. 111 patients met the criteria for the diagnosis of cardiac cachexia. Median age was 78(70–83) years. Patients with cardiac cachexia showed higher age [79 (73–84) vs. 77 (68–82) years; P = 0.005], length of hospital stay [12 (8–15) vs. 9 (6–13) days; P = 0.004], and RDW [14.9 (13.9–16.3) vs. 15.3 (14.3–16.9); P = 0.02] with respect to patients without cachexia. Moreover, patients with cachexia demonstrated reduced eGFR [53 (38–68) vs. 48 (31–60) ml/min/m2; P = 0.03] and TAPSE [18 (15–20) vs. 15 (14–19) mm; P = 0.002] compared to patients without cachexia. No differences were found among groups in terms of NTproBNP. In-hospital mortality was higher in patients with cachexia compared to other patients (6.3% vs. 1.3%; P = 0.005). Univariate Cox regression analysis confirmed the poor prognosis of patients with cachexia at one month [HR: 2.53 (1.24–5.19); P = 0.01], six months [HR: 2.47 (1.61–3.77); P < 0.001] and 1 year [HR: 2.04 (1.40–2.98); P < 0.001]. Conclusions Patients with cardiac cachexia were characterized by renal dysfunction and right ventricle dysfunction. These alterations should act as worsening factors in terms of abdominal venous congestion and subsequent malabsorption. Finally, in our population, cardiac cachexia was related to poor short term and long term outcome as confirmed by recent studies.

Cold-induced phase separation for the simple and reliable extraction of sex hormones for subsequent LC-MS/MS analysis

Sex hormones, including androgens, estrogens, and progestogens, are important biomarkers for various diseases. Quantification of sex hormones is typically conducted by LC-MS/MS. At present, most methods require liquid-liquid extraction or solid phase extraction for sample preparation. However, these pretreatments are prone to compromise LC-MS/MS throughput. To improve on the current standard practices, we investigated cold-induced phase separation for sex hormone extraction. After protein precipitation with acetonitrile and adjusting the solution constitution with water, samples were stored at −30°C for 10 min to generate two distinct phases: an acetonitrile-rich layer on top of a water-rich layer. During this process, the hydrophobic sex hormones spontaneously separate into the upper layer. This simple and reliable cold-induced phase separation-based LC-MS/MS methodology was used here to simultaneously detect estrone, estradiol, estriol, testosterone, androstenedione, dehydroepiandrosterone, progesterone, and 17-hydroxyprogesterone in serum. Validation of this method indicated satisfactory performance, including acceptable linearity, accuracy, precision, and tractability. Compared with the mainstream liquid-liquid extraction-based method, this new method exhibits significant progress in throughput, which shortens the time cost of sample preparation from 90 to 40 min. We propose that this method can be an excellent alternative for sex hormone analysis in routine clinical laboratories.

The influence of social isolation on the incidence of positivity in covid-19 tests in a metropolitan region of São Paulo, Brazil

Introduction: With the arrival of the SARS-CoV-2 (Coronavirus 2 of severe acute respiratory syndrome) pandemic in Brazil, especially in the city of São Paulo, there was a need to apply social isolation policies associated with testing, covering all municipalities. The Clinical Analysis Laboratory of Centro Universitário FMABC was one of the first laboratories to receive certification and qualification to perform RT-PCR (reverse transcriptase reaction followed by polymerase chain reaction) tests in the metropolitan region of São Paulo. Objective: Aim to analyze the influence of adopting social isolation on the incidence of positivity in COVID-19 tests in the metropolitan region of São Paulo, Brazil. Methods: a descriptive study carried out from March to May 2020, epidemiological data were collected from each unit served and organized by the data controllership team of the Clinical Analysis Laboratory of FMABC. Epidemiological, demographic, and laboratory data were extracted from the Matrix® outpatient data management system. Clinically suspected cases and confirmed by laboratory tests (RT-PCR and serological tests) were entered. The tests were divided into serological tests using the RT-PCR molecular test, on samples of nasopharyngeal mucosal scrapings collected with sterile Swab. Results: It were evaluated PCR test and antibody presence (IgA, IgM and IgG) in blood samples of 16.297 patients. 22.718 tests were performed for the diagnosis of COVID-19, both RT-PCR (10.410 tests) and serological tests to detect anti-SARS-CoV-2 antibodies, IgA, IgM and IgG, a total of 16.297 patients were assessed, 63% women and 37% men. It was observed that the social isolation policies adopted during this period contained the massive expansion of contamination, at least while the social isolation rates were above 55%. Conclusion: The data of this study demonstrated the effectiveness of social isolation in containing the positive contamination of SARS-CoV-2 in the metropolitan region of São Paulo, at least for the first three months.

ANALYSIS OF REGISTERED PATIENTS DIAGNOSED WITH GONORRHEA IN BUKHARA REGION AND A METHOD FOR COMPARING THE EFFECTIVENESS OF THE DRUGS USED DEPENDING ON THE CLINICAL COURSE OF THE DISEASE

The research was conducted in 76 patients with gonorrhea aged 14 to 60 years. The diagnosis of gonorrhea was confirmed by clinical, epidemiological and laboratory methods. It has been studied what antibacterial drugs patients take to achieve good results. Based on the obtained results, the indicators were statistically processed. Which antibiotics are significantly more susceptible to gonorrhea have been studied and determined. In the acute, subacute, chronic type of gonorrhea on clinical course, the use of the antibiotic Cefakim + Azithromycin has shown reliable results in clinical course and laboratory analysis.

Anticoagulant Activity of Crude and Phenolic Extracts of Dalbergia ecastaphyllum (L.) Taub. Dried Leaves

Pharmacognosy Research,2021,13,3,121-128.DOI:10.5530/pres.13.3.3Published:June 2021Type:Original ArticleAuthors:Enrico Rogatto Rovetta, Mara Angelina Galvão Magenta, Thaís Moura Gascon, Fernando Luiz Affonso Fonseca, Robson Miranda da Gama, José Armando Jr, and Horácio Dorigan Moya Author(s) affiliations:Enrico Rogatto Rovetta1, Mara Angelina Galvão Magenta2, Thaís Moura Gascon3, Fernando Luiz Affonso Fonseca3, Robson Miranda da Gama1, José Armando Jr1, Horácio Dorigan Moya1,* 1Centro Universitário FMABC, Pharmacy Laboratory, Santo André, São Paulo, BRAZIL. 2Universidade Santa Cecília, Santos, São Paulo, BRAZIL. 3Centro Universitário FMABC, Clinical Analysis Laboratory, Santo André, São Paulo, BRAZIL. 4Centro Universitário FMABC, Analytical Chemistry Laboratory, Santo André, São Paulo, BRAZIL. Abstract:Background: Ischemic diseases are one of the leading causes of death worldwide; therefore, the search for new compounds with safer and more effective anticoagulant and antiplatelet action becomes relevant. Objectives: To evaluate the impact of Dalbergia ecastaphyllum (L.) Taub. dry leaf extract, a species of the Fabaceae family that is commonly found in Brazil, on blood coagulation. Materials and Methods: A statistical design of experiments was carried out with selection of five factors and two levels, resulting in 32 different ethanolic raw extracts. The extracts were resuspended in saline and tested in a pool of human blood. The activated partially thromboplastin time (APTT) and the prothrombin time (PT) were analysed in a Thrombolyzer®. Results: Treatment using ethanol 70%, a drug/solvent ratio of 5%, pH 9 and dynamic maceration for 6 hrs resulted in the best combination for the extraction of total phenols, while ethanol 70%, a drug/solvent ratio of 5% and dynamic maceration for 24 hrs showed the best flavonoid extraction, with the pH being inconclusive. On average, all extracts were capable of increasing both APTT and PT, especially those in lower dilutions (1:5); however, the overall efficacy of the extracts is inconclusive. Conclusion: The species under study showed important results in the blood clotting process in vitro, increasing the mean time of APTT and PT, which suggests that Dalbergia ecastaphyllum (L.) Taub. has some potential as a possible drug in the anticoagulant class. Keywords:Anticoagulant activity, Dalbergia ecastaphyllum (L.) Taub., Flavonoids, Phenolic compounds, Plant extractsView:PDF (903.36 KB)

Pathogenesis of IgA Nephropathy: Current Understanding and Implications for Development of Disease-Specific Treatment.

IgA nephropathy, initially described in 1968 as a kidney disease with glomerular “intercapillary deposits of IgA-IgG”, has no disease-specific treatment and is a common cause of kidney failure. Clinical observations and laboratory analyses suggest that IgA nephropathy is an autoimmune disease wherein the kidneys are damaged as innocent bystanders due to deposition of IgA1-IgG immune complexes from the circulation. A multi-hit hypothesis for the pathogenesis of IgA nephropathy describes four sequential steps in disease development. Specifically, patients with IgA nephropathy have elevated circulating levels of IgA1 with some O-glycans deficient in galactose (galactose-deficient IgA1) and these IgA1 glycoforms are recognized as autoantigens by unique IgG autoantibodies, resulting in formation of circulating immune complexes, some of which deposit in glomeruli and activate mesangial cells to induce kidney injury. This proposed mechanism is supported by observations that (i) glomerular immunodeposits in patients with IgA nephropathy are enriched for galactose-deficient IgA1 glycoforms and the corresponding IgG autoantibodies; (ii) circulatory levels of galactose-deficient IgA1 and IgG autoantibodies predict disease progression; and (iii) pathogenic potential of galactose-deficient IgA1 and IgG autoantibodies was demonstrated in vivo. Thus, a better understanding of the structure–function of these immunoglobulins as autoantibodies and autoantigens will enable development of disease-specific treatments.

Microbiotes of the vagina, cervical canal and uterine cavity in chronic endometritis

The objective: to determine the profile of vaginal, cervical and uterine microbiota in chronic endometritis in patients with failed ART attempts in history. Materials and method. A prospective study and clinical laboratory analysis of vaginal microbiocenosis, cervical canal and uterine cavity in women with chronic endometrite in a history on the basis of MC LLC «ISIDA - IVF», in the period from 2020 to 2021 was carried out. The group consisted of 55 women of reproductive age with chronic endometritis and repeated failed ART attempts, and 40 women of reproductive age without fertility impairment who formed the control group. Amount of research – Vaginal Secretion PH, Vaginal Microbial Estimation of Pap Smear Microscopy, Polymerase Chain Reaction Study (PCR), Culture Study of Vaginal Secretions, Study of microflora of the cervical channel and bioptat of endometrium. Results. In women with infertility, against the background of chronic endometritis in history, as a result of bacterial sowing from the uterine cavity, the predominance of mixed bacterial flora, which is represented by various associations of opportunistic and pathogenic micro-organisms, has been detected. Chronic inflammatory endometry is caused by a persistent viral bacterial infection in the uterine cavity, which differs in species composition from the microflora of the lower genital tract. Conclusions. The characteristic feature of the vaginal flora in chronic endometritis is a high level of dissbiotic changes, combined with high values of the notional pathogenic flora. The microbiotopes of the cervical channel and endometrium in the inflammatory processes of endometry are characterized by the predominance of obligatory-anaerobic micro-organisms in the form of associations with viral infection much more than twice as frequently detected in endocervix as compared to endometrium.

917P Head and neck cancer patients under (chemo-)radiotherapy undergoing nutritional intervention: Results from the prospective randomized HEADNUT-trial

Patients with head and neck squamous cell carcinoma (HNSCC) are often malnourished before starting the primary or adjuvant radiotherapy (RT) or chemoradiotherapy (CRT). Due to therapy related adversary effects of the RT or CRT the number of malnourished patients rises to 88% during ongoing therapy. Malnutrition can have a negative influence on therapycourse and clinical outcome. There are still not enough prospective, randomized studies on the influence of nutritional status on adversary effects of (C-)RT during therapy. From October 2018 to October 2020, the nutritional status of 61 patients with HSNCC, randomized into a control and intervention group (IG), was measured at the beginning, every 2 weeks during and at the end of therapy. The risk of malnutrition was evaluated with established malnutrition screening tools (NRS-2002, MUST, Nutriscore) and 3-day diet diaries, patients underwent clinical examinations, laboratory analyses and bioelectrical impedance analyses. In addition, the IG had individualized nutritional counseling every 2 weeks. Before therapy, the median BMI of all patients was 23.8 (14.5-37.2) kg/m2 and after therapy median BMI fell to 22.9 (16.8-33) kg/m2 (p<0.001). The baseline median fat-free mass index (FFMI) for all patients was 18.1 (14-24.7) kg/m2, which dropped to 17.8 (13.4-21.6) kg/m2 at the ending of therapy (p<0.001). Control group patients lost more weight compared to compliant intervention group participants with a BMI <22 kg/m2(p=0.0015, CI: 0.33-2.95). MUST was the only questionnaire, which showed high sensitivity (86%) and specificity (88%) at baseline in predicting malnutrition. The 2-year overall survival rate was 70% in the control group 79% in the intervention group (log-rank p=0.79). A clear impact on the overall survival rate was observed in pretherapeutic phase angle, posttherapeutic FFMI and albumin level in blood as indicators (log-rank p=0.002, p=0.008, p=0.016). Malnutrition has an adverse impact on patients’ outcome undergoing (chemo-) radiotherapy. Pretherapeutic phase angle, posttherapeutic FFMI and albumin level can be considered as indicators for overall survival.

Food intake, and anthropometrical and biological parameters in adult Tunisians during Ramadan

We evaluated the effects of fasting during Ramadan on nutritional intake and plasma lipoproteins in 20 healthy adults of normal weight. A 5-day food questionnaire was completed for every participant. Clinical investigations, anthropometrical measurements and laboratory analysis were also undertaken. Body weight, blood pressure and blood glucose were not influenced by fasting but there were non-significant modifications in the plasma lipid fractions. The total cholesterol remained unchanged. Total daily energy intake was comparable before, during and after Ramadan despite the decrease in meal frequency during fasting. Thus fasting in Ramadan did not affect dietary intake, clinical, anthropometrical and most biological parameters

Characterization of the dog urine microbiota and its susceptibility to antimicrobials

The urinary tract of dogs is usually considered to be a sterile environment, with the exception of the distal urethra, which is composed of resident microorganisms from the vagina or foreskin. Approximately 14% of dogs can develop urinary tract infection (UTI) during their lifetime, being more common in females and the elderly. The indiscriminate use of antimicrobials in the treatment and prevention of diseases of the urinary tract can contribute to the development of bacterial resistance. The aim of this study was to identify the microbiota involved in disorders of the urinary tract of dogs in the municipality of Formiga, Minas Gerais and to establish the susceptibility profile of these agents against antimicrobials commonly used in the veterinary clinical routine. The samples used were granted by a veterinary clinical analysis laboratory in the municipality of Formiga - MG, with 13 biological samples being made available and submitted to culture and antibiogram. In eight (61.5%) of the 13 samples, it was possible to notice the presence of Staphylococcus spp., followed by E. coli (15.4%), Enterococcus sp. (7.7%), Streptococcus sp. (7.7%) and Proteus sp. (7.7%). Regarding the susceptibility profile to antimicrobials, it was possible to conclude that for amoxicillin + clavulanic acid, with approximately 61.5% of the samples, it was the antimicrobial with the best efficiency.

MM-340: Plasma Cell Neoplasm in the Central Nervous System: A Case Report

Context Plasma cell neoplasms are a heterogeneous group of diseases involving clonal plasma cell production, secreting monoclonal proteins. Usually, these are conditions that occur in the bone marrow, and involvement in the central nervous system (CNS) is a rare event, affecting about 1% of patients and with unfavorable prognosis. Early diagnosis is indispensable for the initiation of appropriate therapy. Objective The present work aims to describe two cases of patients with plasma cell neoplasms with infiltration in the CNS. Design Case report. Setting Case report with emphasis on laboratory diagnosis performed in a private laboratory of clinical analysis. Patients or Other Participants Case 1: Male patient, 58 years old, previous diagnosis of monoclonal gammopathy IgA, previous plasmocytoma in the sternal region, recurrence with orbital and sacral plasmocytoma treated with cyclophosphamide, thalidomide, and dhimhasone, and refused autologous bone marrow transplantation. He was hospitalized for seizure investigation. Case 2: A 51-year-old male patient with a previous diagnosis of MM; after autologous bone marrow transplantation, he sought medical service with a febrile peak 10 months after transplantation. Clinical evaluation and imaging tests showed multiple sparse lytic lesions in the vertebrae and other permeative bodies distributed throughout the skull cap. Presented with headache and visual turbidity. Interventions Case 1: The CSF sample was collected for cytological analysis, which showed 65% of cells with aberrant plasma cells. Immunophenotypic analysis demonstrated the expression of CD38++ CD138+ plasma markers, sIg-kappa+ cytoplasmic light chain restriction and phenotypic aberration. Case 2: Cytologic analysis detected 95% of aberrant plasmocytes and CFS immunophenotyping, presenting 94.4% of lambda monoclonal plasmocytes. Results Case 1: The patient underwent intrathecal treatment; however, there was progressive worsening and death. Case 2: Polychemotherapy was performed, and the patient had a satisfactory clinical response. Conclusions Clinical symptoms related to the involvement of the CNS in plasma cell neoplasms can be confused with symptoms due to hypercalcemia, hyperuremia, and paraproteinemia, making the laboratory analysis of CSF of paramount importance for proper diagnosis. Pathological cells have aberrant cytological characteristics; therefore, confirmation through immunophenotyping is essential, being a sensitive and quick-to-perform test for diagnostic confirmation. Plasma cell neoplasms are a heterogeneous group of diseases involving clonal plasma cell production, secreting monoclonal proteins. Usually, these are conditions that occur in the bone marrow, and involvement in the central nervous system (CNS) is a rare event, affecting about 1% of patients and with unfavorable prognosis. Early diagnosis is indispensable for the initiation of appropriate therapy. The present work aims to describe two cases of patients with plasma cell neoplasms with infiltration in the CNS. Case report. Case report with emphasis on laboratory diagnosis performed in a private laboratory of clinical analysis. Case 1: Male patient, 58 years old, previous diagnosis of monoclonal gammopathy IgA, previous plasmocytoma in the sternal region, recurrence with orbital and sacral plasmocytoma treated with cyclophosphamide, thalidomide, and dhimhasone, and refused autologous bone marrow transplantation. He was hospitalized for seizure investigation. Case 2: A 51-year-old male patient with a previous diagnosis of MM; after autologous bone marrow transplantation, he sought medical service with a febrile peak 10 months after transplantation. Clinical evaluation and imaging tests showed multiple sparse lytic lesions in the vertebrae and other permeative bodies distributed throughout the skull cap. Presented with headache and visual turbidity. Case 1: The CSF sample was collected for cytological analysis, which showed 65% of cells with aberrant plasma cells. Immunophenotypic analysis demonstrated the expression of CD38++ CD138+ plasma markers, sIg-kappa+ cytoplasmic light chain restriction and phenotypic aberration. Case 2: Cytologic analysis detected 95% of aberrant plasmocytes and CFS immunophenotyping, presenting 94.4% of lambda monoclonal plasmocytes. Case 1: The patient underwent intrathecal treatment; however, there was progressive worsening and death. Case 2: Polychemotherapy was performed, and the patient had a satisfactory clinical response. Clinical symptoms related to the involvement of the CNS in plasma cell neoplasms can be confused with symptoms due to hypercalcemia, hyperuremia, and paraproteinemia, making the laboratory analysis of CSF of paramount importance for proper diagnosis. Pathological cells have aberrant cytological characteristics; therefore, confirmation through immunophenotyping is essential, being a sensitive and quick-to-perform test for diagnostic confirmation.

EVALUATION OF A NOVEL LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY BASED METHOD FOR VITAMIN D ANALYSIS IN BLOOD

Objective: To evaluate a novel liquid chromatography tandem mass spectrometry based method for serum 25 hydroxy vitamin D (D2 and D3 metabolites) analysis.&#x0D; Study Design: Cross sectional validation study.&#x0D; Place and Duration of Study: Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology, Pakistan, from Mar 2019 to Mar 2020.&#x0D; Methodology: Samples were extracted and 25 OH vitamin-D was separated by means of chromatography and finally quantified via mass spectrometer. A quadrupole- tandem mass spectrometer with Electron spray Ionization coupled to high performance liquid chromatography was adopted for detection and quantitation of 25-hydroxyvitamin D2 and D3 in serum.&#x0D; Results: Limit of detection (LOD) was at the level of 2.49 ng/ml and limit of quantitation (LOQ) was estimated to be 3.9 ng/ml for both the metabolites. The correlation coefficient was 0.99. For D3 observed recovery was 98% and 97.5% respectively while for D2 the recovery was calculated to be 97% and 98%. Percentage relative standard deviation (RSD) was 0.8% and 1.3% respectively. This method has an advantage of less matrix effects, minimal cross-reactivity with 24, 25 hydroxy vit D and 25, 26 di-hydroxy vitamin D metabolite than the routinely used antibody-based assays.&#x0D; Conclusion: This LC-MS/MS methodology is sensitive, specific and can quantitate Vitamin D2 and D3 quite efficiently. This method may be employed for vitamin D analysis in clinical laboratories.

RELATIONSHIP BETWEEN ORAL MELANOTIC HYPERPIGMENTATION AND SERUM INFLAMMATORY CYTOKINES IN HIV SEROPOSITIVE PATIENTS—A CASE CONTROL STUDY

<h3>Background</h3> Oral melanotic hyperpigmentation in patients with HIV (HIV-OMH) has been attributed to the use of antifungal agents or antiretroviral drugs; alternatively, HIV-induced cytokine dysregulation causing activation of melanogenesis pathway has been well documented. <h3>Objective</h3> To assess the level of cytokine dysregulation as a possible biological etiologic factor for oral melanotic hyperpigmentation in patients newly diagnosed with HIV. <h3>Methods</h3> A case-control study conducted among newly diagnosed HIV seropositive patients yet to commence highly active antiretroviral therapy. Cases were patients with HIV with OMH, and the control group included age- and sex-matched patients with HIV without OMH. Clinical features and laboratory analysis of CD4 count and cytokine levels (IL-6 and tumor necrosis factor alpha) were compared between the OMH and non-OMH groups. <h3>Results</h3> Participants' ages ranged from 22 to 68 years, with a mean of 42.4 ± 10.7 years. The tongue 34 was the most commonly affected site (75.5%), and multiple-site involvement was seen in 9 cases (25.7%). The mean rank value of tumor necrosis factor alpha was slightly higher among cases (36.24 pg/mL) compared to controls (34.76 pg/mL). On the other hand, the mean rank value of IL-6 was lower among cases (33.93 pg/mL) compared to controls (37.07 pg/mL). Thirty-three cases (94.3%) and 23 controls (65.7%) had CD4 count ≤350 cells/mm³ (<i>P</i> = .003). <h3>Conclusions</h3> There was no statistically significant difference in the serum cytokine levels of those with HIV-OMH and those without it. There was a statistically significant relationship between HIV-OMH and severe immunosuppression.

Analytical toxicology service model at the subregional center level for severe acute poisoning

Acute poisoning may necessitate identification of the toxic agent; however, several acutely poisoned patients are treated with minimal laboratory assistance. We investigated whether focused reference to laboratory toxicology tests conducted during a pilot project for a subregional analytical toxicology service influences treatment decisions. Patients with acute poisoning presented to the level 1 regional emergency medical center from May 2018 to April 2019 were initially reviewed. Poison samples were referred to the subregional toxicological analytical service. In total, 253 substance samples were tested among 111 patients during the study. According to the reported drug levels, 3 (1.2%) samples contained lethal doses, 49 (19%) had toxic levels, and 28 (11%) contained detectable levels of a lethal toxin or pesticide. Disagreement between the clinical assessment and laboratory analyses was found for 62 patients (fair kappa = 0.24, 56%), and they often had lower Glasgow Coma Scale, higher severity scores, older age, and less likelihood of receiving gastrointestinal decontamination. The regional analytical toxicology services were helpful for diagnostic planning and therapeutic management of acute poisoning. For seriously poisoned patients with inconsistent histories, it is necessary to reevaluate the classic therapeutic process based on the medical history.

Differential diagnosis of bovine intestinal diseases and their sequelae regarding ultrasonography and other diagnostic tools.

Background and Aim:Intestinal disorders represented a large proportion of abdominal emergencies in bovine practice, and their definite diagnosis was a big challenge. The study described different intestinal disorders in cattle either in the small intestine (SI) or large intestine with their sequelae and peritonitis between SI loops.Materials and Methods:This study involved healthy (n=20) and diseased (n=40) cattle with intestinal disorders. All animals were undergoing clinical examination, laboratory analyses, and ultrasonographic examination.Results:Diseased cattle had monocytic leukocytosis as well as hypoproteinemia and hypoalbuminemia. The SI ileus of either proximal (n=12) or distal (n=15), intussusception (n=3), cecal, and/or colonal dilatation (n=10) were diagnosed by ultrasound and other diagnostic tools. Duodenum intussusception was imaged in cross-section as bull’s eyes lesions. Animals with SI obstructions showed complete cessation (ileus with complete obstruction) or partial reduction of the peristaltic SI movement (ileus with partial obstruction), dilated duodenum (6.5-9.9 cm), and dilated jejunum and/or ileum (4.4-6.8 cm). Ultrasonography diagnosed SI ileus, due to either intestinal obstruction or peritonitis, and detected the ileus site, either proximal or distal. Cecal/colonal dilatation was detected using ultrasonography in which SI was not imaged and the peristaltic movements were completely reduced. The recorded intestinal disorders were associated with other disorders (e.g., liver cirrhosis or peritonitis).Conclusion:Ultrasonography played an important role in the differential diagnosis of intestinal disorders in cattle. Peritonitis between SI loops and cecal and/or colonal dilatation was also diagnosed.

Innovative electrochemical sensor for the precise determination of the new antiviral COVID-19 treatment Favipiravir in the presence of coadministered drugs.

Due the current pandemic of COVID-19, an urgent need is required for serious medical treatments of a huge number of patients. The world health organization (WHO) approved Favipiravir (FAV) as a medication for patients infected with corona virus. In the current study, we report the first simple electrochemical, greatly sensitive sensor using MnO2-rGO nanocomposite for the accurate determination of Favipiravir (FAV). The developed sensor showed a high improvement in the electrochemical oxidation of FAV comparing to the unmodified screen-printed electrode (SPE). The suggested platform constituents and the electrochemical measurements parameters were studied. Under optimal experimental parameters, a current response to the concentration change of FAV was found to be in the linear range of 1.0 × 10-8-5.5 × 10-5 M at pH 7.0 with a limit of detection 0.11 µM and a quantification limit of 0.33 µM. The developed platform was confirmed by the precise analysis of FAV in real samples including dosage form and plasma. The developed platform can be applied in different fields of industry quality control and clinical analysis laboratories for the FAV determination.

Cervical Cancer Screening by Association of Liquid-Based Cytology with Real-Time PCR for High-Risk Human Papillomavirus in Women Assisted in a Clinical Analysis Laboratory

Cervical Cancer Screening by Association of Liquid-Based Cytology with Real-Time PCR for High-Risk Human Papillomavirus in Women Assisted in a Clinical Analysis Laboratory