Sediment Smears Research Articles

Detection of Bk Polyomavirus Antigen in Patients with Acute Lymphoblastic Leukemia

Background: BK polyomavirus that is acquired in early childhood as BKPyV is near ubiquitous in adults with a seroprevalence of >80%, it has been found as the main cause of hemorrhagic cystitis in hematopoietic stem cell transplantation patients, due to immunosuppression regimen that lead to the activation of the virus from the latency status and lead to increased viral shedding in urine (viuria).
 Objective: To investigate the frequency of BK Polyomavirus antigen excretion in urine of children with acute lymphoblastic leukemia with and without chemotherapy and compare it with normal controls. 
 Patients and Methods: A case-control study conducted from December 2021 to May 2022 in Baghdad, Iraq on leukemia patients of Central Pediatrics Hospital (Al-Eskan ). Urine samples and urine sediment smears were collected from 60 acute lymphoblastic leukemia patients. And compared with 60 apparently healthy age and sex-matched children, BK polyomavirus antigen in urine was detected using Enzyme-linked Immunosorbent Assay and urine cytology were Pap stained for the detection of decoy cells (DCs).
 Results: Positive BKPyV antigen in urine was seen in 55 (91.7%) of Acute lymphoblastic leukemia patients and 39 (65.0%) in controls (p=<0.001) and all the patients were decoy negative. There was no significant effect of the positivity of antigenuria on neither leukocytes level nor on the occurance of relapse in leukemia patients. 
 Conclusion: The very high frequency of BKPyV in the urine signifies the importance of reactivation of this virus in ALL patients with and without chemotherapy.

Detection of Bk Polyomavirus Antigen in Patients with Acute Lymphoblastic Leukemia

Background: BK polyomavirus that is acquired in early childhood as BKPyV is near ubiquitous in adults with a seroprevalence of >80%, it has been found as the main cause of hemorrhagic cystitis in hematopoietic stem cell transplantation patients, due to immunosuppression regimen that lead to the activation of the virus from the latency status and lead to increased viral shedding in urine (viuria). Objective: To investigate the frequency of BK Polyomavirus antigen excretion in urine of children with acute lymphoblastic leukemia with and without chemotherapy and compare it with normal controls. Patients and Methods: A case-control study conducted from December 2021 to May 2022 in Baghdad, Iraq on leukemia patients of Central Pediatrics Hospital (Al-Eskan ). Urine samples and urine sediment smears were collected from 60 acute lymphoblastic leukemia patients. And compared with 60 apparently healthy age and sex-matched children, BK polyomavirus antigen in urine was detected using Enzyme-linked Immunosorbent Assay and urine cytology were Pap stained for the detection of decoy cells (DCs). Results: Positive BKPyV antigen in urine was seen in 55 (91.7%) of Acute lymphoblastic leukemia patients and 39 (65.0%) in controls (p=<0.001) and all the patients were decoy negative. There was no significant effect of the positivity of antigenuria on neither leukocytes level nor on the occurance of relapse in leukemia patients. Conclusion: The very high frequency of BKPyV in the urine signifies the importance of reactivation of this virus in ALL patients with and without chemotherapy. Keywords: Acute lymphoblastic leukemia, BK Polyomavirus, Antigenuria

Bronchoalveolar Lavage Cytology in Severe Equine Asthma: Cytocentrifugated versus Sediment Smear Preparations.

Equine asthma is a common respiratory disease that may affect horses of any age. The diagnosis of severe equine asthma (SEA) (historically referred as recurrent airway obstruction or RAO) is based mainly on the history of the animal and clinical signs, which are further supported by the cytological examination of the bronchoalveolar lavage (BAL). This can also be helpful in monitoring the inflammation of the lower airways in response to environmental management and medication. The cytocentrifugated preparation is usually considered the method of choice for BAL cytological interpretation. The aim of this study was to compare the results in terms of differential cell counts (DCC) in BAL cytology performed on sedimented smears and cytocentrifugated preparations. To carry this out, 48 BAL samples were collected from six horses with SEA that were subjected to a process of exacerbation of the disease by environmental stimuli, which was later followed by the appropriate treatment. Each collected BAL fluid was equally divided into duplicate portions: one processed by cytocentrifugation and one by sediment smear from simple centrifugation. Cytologic examination of all BAL by both methods showed poor concordance in DCC, although it was still able to allow diagnostic recognition of severe lung neutrophilic disorders. These results suggest that sediment smear preparation, although remaining a useful method in general equine practice associated with clinical assessments in the diagnosis of SEA under conditions where there is no possibility of using a cytocentrifuge, cannot be considered a comparable alternative.

The value of Xpert MTB/RIF assay of urine samples in the early diagnosis of smear-negative urinary tuberculosis

BackgroundAccording to reports, between 30 and 40 percent of extrapulmonary tuberculosis (EPTB) cases are caused by urinary tract tuberculosis (UTB). It is critical to identify UTB quickly since it frequently precedes delayed medical attention, which can have detrimental effects. This study examined the use of Xpert MTB/RIF, a PCR test that can detect MTB as well as resistance to an important drug, rifampicin (RIF), in UTB particularly, for the early identification of UTB.Methods180 participants with clinically presumptive UTB whose urine samples were chosen for urine sediment smear, culture, Xpert MTB/RIF, and TB-DNA testing at Henan Chest Hospital between January 2019 and July 2022. Evaluation of test performance using Composite Reference Standards (CRSs). We studied and compared the positivity rate for various tests using the t-test. The effectiveness of smear, culture, Xpert MTB/RIF, and TB-DNA was assessed using McNemar test.ResultsIn this subject, a total of 108 participants were diagnosed with UTB, and the positivity rate was 67.1%. Compared with CRS, the positivity rate of Xpert MTB/RIF, smear, culture, and TB-DNA was 29.69% (19/64, P < 0.001), 7.56% (9/119, P < 0.1), 12.12% (4/33, P > 0.05), and 18.75% (6/32, P < 0.1), respectively. The sensitivity of Xpert MTB/RIF assay was significantly better than that of smear and culture tests (78.9% vs. 77.8%, P < 0.05; 78.9% vs. 75%, P < 0.05). Under CRS, the positivity rate for Xpert, culture, and TB-DNA was 31.6% (6/19, P < 0.1), 6.2% (1/16, P > 0.05), and 26.7% (4/15, P > 0.05) for TB-DNA, respectively, compared to smear negative. Xpert MTB/RIF assay specificity was significant for culture and TB-DNA (53.6% vs. 25%, P < 0.01; 53.6% vs. 38.9%, P < 0.05), and Xpert MTB/RIF assay FPV was significant for culture and TB-DNA (53.6% vs. 0%, P < 0.001; 53.6% vs. 0%, P < 0.001).ConclusionXpert MTB/RIF outperforms smear, cultures, and TB-DNA in detecting UTB, plus Xpert MTB/RIF is better suited for early diagnosis in smear-negative UTB.

Urine cytological study in patients with clinicopathologically confirmed neuronal intranuclear inclusion disease.

ObjectiveThe diagnosis of neuronal intranuclear inclusion disease (NIID) is currently based on CGG repeat expansion in the 5′UTR of the NOTCH2NLC gene, or p62-positive intranuclear inclusions in skin biopsy. The purpose of this study is to explore the value of non-invasive pathological findings in urine sediment cells from NIID patients.Materials and methodsTen patients with clinically suspected NIID were enrolled for skin biopsy and gene screening. Morning urine (500 ml) was collected from each patient, and cell sediment was obtained by centrifugation. Urine cytology, including Giemsa staining, p62 immunostaining, and electron microscopic examination, were conducted on cell sediment.ResultsThe main clinical symptoms of 10 patients included episodic disturbance of consciousness, cognitive impairment, tremor, limb weakness, and so on. Cerebral MRI showed that 9 patients had linear DWI high signal in the corticomedullary junction. Genetic testing found that the number of CGG repeat ranged from 96 to 158 in the NOTCH2NLC gene. Skin biopsy revealed that all patients showed p62-positive intranuclear inclusions in 18.5 ± 6.3% of the duct epithelial cells of sweat gland. In contrast, urine sediment smears revealed that only 3 patients had p62 positive intranuclear inclusions in 3.5 ± 1.2% of the sedimentary cells. Ultrastructural examinations showed that intranuclear inclusions were also identified in the cell sediment of the 3 patients.ConclusionUrine cytology may be a new and non-invasive pathological diagnosis technique for some NIID patients, although the positive rate is not as high as that of skin biopsy, which is a sensitive and reliable pathological method for NIID.

Molecular Diagnostics of Lung Cancer in Serous Effusion Samples

For molecular diagnostics of lung cancer samples, often only a small amount of material is available. The ever-increasing number of biomarker testing is in contrast to the amount of material obtained. In that case, cytological specimens, such as serous effusion samples, are one possible option. Effusion samples were prepared as sediment smears or cytospins or as a cell block if needed. Suitable tumor cells areas were marked by a cytopathologist and used for molecular diagnostics, including fast track analysis, parallel sequencing, and/or fluorescence in situ hybridization. In 62 cases of malignant effusion with cells of pulmonary adenocarcinoma, molecular diagnostics were carried out. A fast-track result with the high-resolution melting method for hotspot mutation of KRAS Exon 2 and EGFR exon 21 and fragment length analysis of EGFR exon 19 was available for 43 out of 47 samples (92%). Parallel sequencing was successful for 56 out of 60 samples (93.3%). In the same period, 108 FISH analyses were performed for MET amplification, followed by ROS1, RET, and ALK translocation analysis. If only a limited amount of tissue/biopsy is available, a malignant effusion is advisable to perform on the molecular diagnostics with a high success rate.

Cytologic comparative analysis between stained and unstained smears of urine sediment in urinary tract pathologies

Aim: To analyse the routine microscopic examination of urine specimens in urinary tract pathologies and compare the cytological examination of unstained and stained urine sediment samples (Papanicolaou and Romanowsky stains) to establish clinicocytological correlation.Material and Method: The present study was conducted among 500 patients coming to either OPD /admitted to CSSH with urinary pathology. In patients with a clinical history suggestive of any urinary pathology freshly voided urine samples were collected. Samples were centrifuged at 1500rpm for 10 minutes. In case of less cellularity, Cytospin at 3000rpm for 5-10 minutes was done. From the sediment, three smears were prepared i.e. smear 1 (routine urine microscopy), smear 2 by Romanowsky stain (Leishman Geimsa) and smear 3 by Papanicolaou method. Results: In the current study, urinary tract pathologies were found to be more common in females (326) as compared to males (174). Majority of patients were in the 3rd and 4th decade of life accounting for 184 cases (38%) and 153 cases (36%) respectively. Stained smears detect high n/c ratio, pus cells, bacilli/fungi and bare nuclei among 95.85%, 99.07%, 99.07% and 0%of UTI samples respectively, while unstained smears detect high n/c ratio, pus cells, bacilli/fungi and bare nuclei in 91.24%, 96.3%, 97.23% and 44.24% of the UTI samples respectively.Conclusion: We can conclude that this study demonstrates the high correlation between the cytological examination of urine specimen in fresh smears and in the Papanicolaou-stained smears.

The advantages of using cytospins of uterine lavage fluid for the diagnosis of equine endometritis

Uterine lavage (UL) is a routine diagnostic procedure for endometritis. In UL the fluid is centrifuged and the sediment smeared. Samples prepared in cytocentrifuges, the so-called “cytospins”, are useful for evaluating cells in fluids, but never been used in UL. The aim of this study was to assess the usefulness of cytospins after UL, comparing automatic versus manual cytocentrifuges, and to determine their value for the diagnosis of endometritis. The study was divided in two parts. Firstly, UL was performed in 16 mares and a small part of the retrieved fluid was cytocentrifuged in an automatic (PreCyto) and manual (PreMan) cytocentrifuge, whereas the remaining fluid was centrifuged. After that, the sediment was divided into three quotas. One quota was smeared, one was processed in an automatic cytocentrifuge (PostCyto) and the last quota was cytospinned in a manual apparatus (PostMan). Cytospins obtained were scored for cellularity, cell preservation, presence of inflammatory cells, bacteria and contaminants; results were compared with sediment smears. Secondly in this study, the best cytospin method was compared with sediment smears in another group of 13 mares, which had endometrial biopsy after UL. Agreement between sediment smears and cytospins was poor to moderate. Compared to sediment smears, cytospins were more cellular, with better morphological details. Urine crystals and fecal contamination were detected more often in cytospins (especially PostCyto and PostMan). No differences in the percentage of inflammatory or epithelial cells existed. PostMan was considered the best method to evaluate UL fluid and it had higher sensitivity (80%), compared with sediment smears (60%), for diagnosing endometritis. Cytocentrifugation offers significant advantages over sediment smears and the manual cytocentrifuge is well suited for horse stable conditions.

Cellular analysis of bronchoalveolar lavage fluid in various lung lesions

Introduction: Bronchoalveolar lavage (BAL) is a standard tool in the diagnosis of lung diseases. Cellular analysis of BAL fluid when deviates from normal are indicative of an inflammatory/infiltrative process. This study involves an analysis of BAL fluid performed manually and also using an automated analyzer which offers a rapid and reliable method for analysis of unprocessed BAL fluid. Aims and Objectives: To utilize the cellular analysis of bronchoalveolar lavage (BAL) fluid in the diagnosis of various lung diseases. Materials and Methods: 37 patients with pulmonary diseases of varying etiologies were included in this study. Fiberoptic bronchoscopy with BAL was performed. BAL fluid was run in SYSMEX XNL/350 six-part analyzer which gave total and differential cell count (mononuclear and polymorphonuclear cells). The sample was also centrifuged, sediment smears prepared and differential cell counts were determined. Results: Total WBC count ranged from 31 cells/µl to 94 cells/µl with the maximum increase seen in asthma. Mononuclear cells were increased in the majority of cases, being 84% in pneumoconiosis with the predominance of macrophages. An increase in lymphocytes was seen in Non-Specific Interstitial Pneumonia (45.3%) and Bronchiolitis Obliterans Organising Pneumonia (35.3%), neutrophils in Usual Interstitial Pneumonia (26.2%) and eosinophils in Chronic Eosinophilic Pneumonia (55.5%) and asthma (8.4%). There were 58% mononuclear cells and 42% polymorphonuclear cells in tuberculosis. There were two cases of adenocarcinoma and one case of squamous cell carcinoma in the present study. Conclusion: BAL cell differentials when combined with additional clinical and radiographic information help secure a confident diagnosis in lung diseases and help clinicians to make therapeutic decisions.

Antineoplastic Chemotherapy and its Effects on the Gastrointestinal Parasitism of the Dog

Background:In veterinary medicine, an increasing incidence of neoplastic diseases has been followed by a growth in the use of chemotherapeutic drugs, often associated with opportunistic infections.Objective:This study aimed to assess the prevalence of gastrointestinal parasites in dogs undergoing antineoplastic chemotherapy in the Lisbon Metropolitan Area, as well as their evolution throughout the protocol and respective effects of chemotherapy on dogs’ lifestyle.Methods:Faecal samples were collected in a group of 30 dogs being treated for neoplastic diseases under different protocols, previous (G1) and during chemotherapy (G2). In total, 60 samples were analysed by Willis flotation, natural sedimentation, Baermann technique and faecal smear stained with Ziehl-Neelsen. A survey to characterize animal lifestyles and deworming care were also conducted with dog’s tutors.Results:In total, there were two positive samples for the protozoanGiardiasp., one of which is in association with the nematodeToxascaris leonina. The two dogs only obtained positive results during chemotherapy (G2). An overall prevalence of gastrointestinal parasitic diseases of 6.7%, in G2, and 0%, in G1, was obtained.Conclusion:The low parasite prevalence has not allowed the evaluation of an association between the use of antineoplastic compounds and infections by gastrointestinal parasites. However, it was concluded that the studied groups were efficiently dewormed, as well as they did not experience any obvious changes in their parasitological component and its lifestyle.

Utility of Cytospin and Cell block Technology in Evaluation of Body Fluids and Urine Samples: A Comparative Study.

Background:Cytologic examination of body fluids commonly involves the use of direct or sediment smears, cytocentrifuge preparations, membrane filter preparations, or cell block sections. Cytospin and cell block techniques are extremely useful in improving cell yield of thin serous effusions and urine samples, and ensure high diagnostic efficacy.Materials and Methods:We studied cytospin preparations and cell block sections prepared from 180 samples of body fluids and urine samples to compare the relative efficiency of cell retrieval, preservation of cell morphology, ease of application of special stains, and diagnostic efficacy. Samples were collected and processed to prepare cytospin smears and cell block sections.Results:We observed that overall, cell yield and preservation of individual cell morphology were better in cytospin preparations as compared to cell blocks, while preservation of architectural pattern was better in cell block sections. The number of suspicious cases also decreased on cell block sections, with increased detection of malignancy. It was difficult to prepare cell blocks from urine samples due to low cellularity.Conclusions:Cytospin technology is a quick, efficient, and cost-effective method of increasing cell yield in hypocellular samples, with better preservation of cell morphology. Cell blocks are better prepared from high cellularity fluids; however, tissue architecture is better studied, with improved rate of diagnosis and decrease in ambiguous results. Numerous sections can be prepared from a small amount of material. Special stains and immunochemical stains can be easily applied to cell blocks. It also provides a source of archival material.

Urine sediment evaluation in the dog and cat

Microscopic examination of the urine sediment is an integral part of urinalysis and a useful, cost-effective diagnostic tool in the everyday practice. Diagnostic investigation of urinary diseases and several systemic metabolic diseases make the microscopic evaluation of urine sediment a priority along with the physical and biochemical examination of urine. The different methods of urine collection (midstream voiding, catheterization or percutaneous cystocentesis) have advantages and disadvantages and the clinician has to decide on the best fitted method of urine collection, according to the medical background of the animal and the case-specific objectives of urinalysis. Proper handling and timely analysis of urine sample are essential for a valid microscopic evaluation. The microscopic examination of urinary sediment is usually conducted in stained or unstained “wet-mounted” preparations; occasionally, air-dried Giemsa-stained sediment smears are examined.Normal urine is sterile and may contain small numbers of cells (white and red blood cells, epithelial cells), few crystals, occasional casts, fat droplets and spermatozoa (male animals). In contrast, large numbers of cells or casts, presence of unusual types of crystals, neoplastic cells, parasites and microorganisms comprise abnormal findings, necessitating a more specialized diagnostic approach. This review focuses on the technical aspects pertaining to the proper sediment microscopic examination, the normally expected elements of sediment and the clinically relevant interpretation of abnormal findings.

Cytokeratin 20 immunocytochemistry on urine sediments: A potential low-cost adjunct to cytology in the diagnosis of low-grade urothelial carcinoma.

Urine cytology is the corner-stone for the diagnosis of urothelial neoplasia; however, a substantial proportion of low-grade carcinomas are reported as inconclusive owing to scant cellularity and subtle cytological features. Biomarkers applied on urine sediment smears of such patients are likely to be clinically relevant. Access to Food and Drug Administration approved urinary biomarkers in resource limited setting is poor. Detection of cytokeratin 20 (CK20) in urine sediments, although still a research tool, is a promising marker as immunocytochemistry is performed regularly in several Indian laboratories. We tested the clinical utility of CK20 immunocytochemistry as a potential low-cost adjunct to urine cytology in diagnosis of low-grade urothelial carcinoma. One hundred and fifty fresh, voided urine specimens from 42 cases of biopsy proven urothelial neoplasia (14 high grade, 28 combined low-grade [n=26]) and low malignant potential [n=2]), and 20 non-neoplastic lesions were included in the study sample. Confident diagnosis of malignancy was possible in five (17.8%) low-grade malignancies. Thirteen of 16 (81.3%) low-grade malignancies with inconclusive cytology showed positive CK20 expression. This reduced the proportion of low-grade cases with inconclusive cytology from 57.1% to 10.7% (P=.021). In addition, we could correctly classify one case of bladder lithiasis with false positive urine cytology. Discrepant CK20 staining (positive) was seen in one patient with acute cystitis. CK20 expression in non-umbrella cells is a robust marker of urinary bladder carcinoma. It has potential clinical utility for identification of low-grade urothelial malignancy with inconclusive cytological diagnosis.

Nationwide cross-sectional survey of schistosomiasis and soil-transmitted helminthiasis in Sudan: study protocol

BackgroundSchistosomiasis and soil-transmitted helminthiasis (STHs) are target neglected tropical diseases (NTDs) of preventive chemotherapy, but the control and elimination of these diseases have been impeded due to resource constraints. Few reports have described study protocol to draw on when conducting a nationwide survey. We present a detailed methodological description of the integrated mapping of schistosomiasis and STHs on the basis of our experiences, hoping that this protocol can be applied to future surveys in similar settings. In addition to determining the ecological zones requiring mass drug administration interventions, we aim to provide precise estimates of the prevalence of these diseases.MethodsA school–based cross-sectional design will be applied for the nationwide survey across Sudan. The survey is designed to cover all districts in every state. We have divided each district into 3 different ecological zones depending on proximity to bodies of water. We will employ a probability-proportional-to-size sampling method for schools and systematic sampling for student selection to provide adequate data regarding the prevalence for schistosomiasis and STHs in Sudan at the state level. A total of 108,660 students will be selected from 1811 schools across Sudan. After the survey is completed, 391 ecological zones will be mapped out. To carry out the survey, 655 staff members were recruited. The feces and urine samples are microscopically examined by the Kato-Katz method and the sediment smears for helminth eggs respectively. For quality control, a minimum of 10% of the slides will be rechecked by the federal supervisors in each state and also 5% of the smears are validated again within one day by independent supervisors.DiscussionThis nationwide mapping is expected to generate important epidemiological information and indicators about schistosomiasis and STHs that will be useful for monitoring and evaluating the control program. The mapping data will also be used for overviewing the status and policy formulation and updates to the control strategies. This paper, which describes a feasible and practical study protocol, is to be shared with the global health community, especially those who are planning to perform nationwide mapping of NTDs by feces or urine sampling.

Prevalence and molecular characterization of Cryptosporidium species in cattle and buffalo calves in Mumbai region of India

Faecal samples of cattle and buffaloes of Mumbai region collected between November 2012 to June 2013 were analysed by conventional and molecular tools to note the prevalence of cryptosporidiosis and species involved in the infection. Conventional analysis viz., direct faecal smear examination, faecal smear examination after normal saline sedimentation, Sheather's floatation and Sheather's floatation sedimentation smear methods demonstrated oocysts of Cryptosporidium in 141 (36.06%) of 391 samples with higher occurrence in buffaloes (36.99%) than cattle (34.48%). Diarrhoeic loose faeces showed higher prevalence (42.07%) than apparently normal faeces (31.72%) irrespective of the host species. When data were arranged as per age groups viz., calves of 0-1month, 1-2months, 2-3months and adults, the highest prevalence was noted in the youngest group (47.12%) declining gradually with the advancing age with lowest (6.25%) in adults indicating inverse correlation between prevalence rate and age of the host. These differences were statistically significant in case of buffaloes. Cryptosporidium andersoni was tentatively identified by morphometric analysis. By employing molecular tools like nested PCR, PCR-RFLP and sequence analysis of few samples showed good correlation in the identification of species of Cryptosporidium involved in the infection and demonstrated occurrence of C. parvum, C. ryanae and C. bovis. Thus all the four commonly occurring bovine species of Cryptosporidium were encountered in the study area which appears to be a first record reporting the occurrence of Cryptosporidium with species level identification in large ruminants from Western region of India. Additionally, the public health significance of C. parvum was also discussed in light of epidemiological factors pertaining to the region.

A non-invasive screening technique for type 1 autoimmune pancreatitis.

IntroductionType 1 autoimmune pancreatitis (AIP) is the pancreatic manifestation of a systemic fibroinflammatory IgG4-related disease. Accurate diagnosis of AIP can avoid major hepatobiliary and pancreatic surgery as it respond dramatically to corticosteroid therapy.AimThis research investigated the feasibility of using peripheral blood cell immunohistochemistry, serum IgG4, T-cell receptor (TCR) and serum isoelectric focusing electrophoresis in the screening of type 1 autoimmune pancreatitis (AIP).Material and methodsThe peripheral blood from 3 type 1 AIP patients, 10 pancreatic cancer patients and 40 normal controls was collected. Sediment smears were jointly incubated with anti-IgG4 and anti-IgG. The percentage of IgG4/IgG positive cells was counted and serum TCR and IgG4 were detected through the whole process. After serum isoelectric focusing electrophoresis, anti-IgG4 and anti-IgG were used to confirm the components of serum.ResultsIn the serum isoelectric focusing electrophoresis, IgG4 and IgG strips showed mirrored distribution in type 1 AIP patients, while there were no strips in the normal controls and pancreatic cancer. Compared with pancreatic tumor patients and healthy controls, serum TCR was significant increased in AIP. The percentage of IgG4/IgG positive cells of peripheral blood cell immunohistochemistry was related to serum IgG4 and hormone therapy reactions.ConclusionsPeripheral blood cell immunohistochemistry, serum IgG4, TCR and serum isoelectric focusing electrophoresis is suitable for the screening of type 1 AIP and monitoring its response assessment.

PEGylation of surfacted magnetite core–shell nanoparticles for biomedical application

The surface of oleate double layer coated (surfacted) magnetite nanoparticles (OA@MNPs) was coated by PEG (poly(ethylene glycol) of Mw=1000, 4000 or 20,000Da, respectively, to get core–shell structured nanomagnets. The oleate bilayers were prepared in two different ways; (i) oleic acid was added directly into the magnetite co-precipitation mixture containing the MNPs to obtain OA@s-MNP samples – s-MNP standing for “as-synthesized MNP” and (ii) sodium oleate (oleate anion, OA) was added to the purified MNPs to obtain OA@p-MNP samples – p-MNP standing for “purified MNP”. The effect of the surfactant addition method on the pH- and ionic strength-dependent stability (dynamic laser light scattering and laser-Doppler electrophoresis experiments), the biomedical applicability (MRI measurements) and the biocompatibility (blood sedimentation and blood smear tests) of the core–shell MNPs was studied. Different mechanisms of oleate adsorption were found in ATR FT-IR experiments (inner sphere surface complexation via ligand exchange for the s-MNPs and additional H-bonding for the p-MNPs), suggesting different behaviour. The colloidal stability and salt tolerance of the two kinds of OA@MNPs were similar, but the hydrodynamic diameter of the OA@s-MNP was considerably larger than that of OA@p-MNP. In accordance with this, the r2 relaxation was also higher for the s-MNP samples (∼400 and ∼200mM−1s−1, respectively). The physico-chemical tests indicate that the OA-coated MNPs form clusters and the degree of clustering of OA@s-MNPs is significantly greater than that of OA@p-MPNs. PEGylation does not appear to affect colloidal stability and salt tolerance meaningfully. The adsorption of PEG was proved experimentally. We have found that the PEG top layer decreases the electrostatic contribution, nevertheless increases the steric contribution of the original electrosteric stabilization caused by the OA double layer. However, an increase in the molecular weight above 1000Da and the amount of added PEG above 5mmol/g gradually reduces the salt tolerance of the samples. The results indicate strong potential for biomedical application and biocompatibility of the PEGylated MNPs.

Microfilariae of Wuchereria Bancrofti in a Patient of Chylous Haematuria: Report of an Unusual Finding in Urine Cytology

Filariasis is a disabling parasitic disease and it constitutes as a major health problem in most of the tropical and sub-tropical countries. One hundred and twenty million people in at least 80 tropical and sub-tropical countries are infected with lymphatic filarial parasite, with one billion people being at risk of the disease. The laboratory diagnosis of filariasis is conventionally made by demonstrating microfilaria in peripheral blood smear. However, microfilaria have been incidentally detected by doing fine needle aspiration of various lesions in clinically unsuspected cases of filariasis. In 1982, detected microfilariae in the sediment smears of cystoscopically catheterized urine samples, but reports on detection of microfilaria in normally voided urine samples are extremely rare. Here, we are reporting, such a case of microfilariae of Wuchereria bancrofti in an 18-year-old boy who presented with chylous haematuria.

Peculiarities regarding the testing of milk physicochemical and cytology at a PIC sows sample.

Testing the main physicochemical and cyto-morphological milk parameters during lactation debut represented the monitoring of breast health at a lot of PIC sows (n = 15) and its impact on health and growth indicators in suckling piglets. Investigations were carried out on samples of milk collected by manual milking, using stimulation of milk ejection by oxytocin injection (n = 10) and by breastfeeding 2-3 piglets (n = 5). Physicochemical tests were performed with semi-automatic EKOMILK analyzer, consisting in determining the main biochemical components (non-fat dry substance - NFS, fat, protein), density, freezing point and water added. Cyto-morphological investigations consisted in making the milk cytograms on the milk sediment smears, defatted with xylene or benzene and panoptic stained (MGG and Dia - Quick panoptic). The evolution of investigated compositional and cytological parameters revealed large variations of individual and average values, grouped in intervals comparable to those reported in consulted studies and with minor deviations and non-pathological connotation. Thus, the test results of milk samples outlined a physicochemical profile, characteristic on lactation debut in the sow, with the following individual oscillations around the average values: 5.37 - 13.3% (7.97%) for protein, 10.8 - 19.9% (13.67%) for non-fat dry substance, 1.71 to 14.5% (8.18%) for fat, 1.028 to 1.076 (1.043) for density, -0.954 to -0.744 °C (0.28 °C) for the freezing point. The cyto-morfological configuration of this sample of sows milk was characterized by increasing the proportion of macrophages with advancing lactation (42.86%, 30-60%), usually associated with PMN leukocytes decreasing (15.8%, 8-43% ) and lymphocytes rarely (31.0%, 11-46%) or epithelial cells (10.4%, 3-20%). We have given a particular character for the onset of lactation in sows to the distribution and morphology of epithelial cells. Regarding the distribution, it was relevant the higher proportion of epitheliocytes (10.4%) in the milk of investigated sows to that found in milk of ruminant species. Most of the observed epitheliocytes had a glandular origin, and morphological were large cells, integral, uni- or binucleate, surrounded by a bulky cytoplasmic mass, low basophilic, with many fat microspheres, often gaining sparkling appearance and rarely ring.

Evaluation of Combined LED-Fluorescence Microscopy and Bleach Sedimentation for Diagnosis of Tuberculosis at Peripheral Health Service Level

BackgroundSputum microscopy is the only diagnostic for tuberculosis (TB) available at peripheral levels of health service in resource-poor countries. Its sensitivity is reduced in high HIV-prevalence settings. Sodium hypochlorite (NaOCl) specimen sedimentation prior microscopy and light-emitting diode (LED)-fluorescence microscopy (FM) can individually improve performance of microscopy. This study aimed to evaluate the performance of combined LED-FM and NaOCl sputum sedimentation for TB detection at peripheral level of health services.MethodsA prospective study was conducted in an urban health clinic in Nairobi, Kenya. Three sputum specimens were collected over 2 days from consecutive TB suspects. Smears were prepared and stained with auramine O and Ziehl-Neelsen (ZN) methods. Bleach (3.5%) was added to the remaining specimen before overnight sedimentation at room temperature. Auramine O staining was performed on smears of sediment. A 4th specimen was collected for TB culture. Auramine smears were read under the same microscope as used for ZN smears, but equipped with the LED FluoLED™ fluorescence illuminator.Results497 patients were included, and 1394 specimens collected. The yield of positive specimen was significantly increased after NaOCl sedimentation (24.9%) compared to direct LED-FM (20.6%) and direct ZN (20.3%). In detecting smear-positive patients, sensitivity was 78.5% for LED-FM after NaOCl sedimentation compared to 73.2% and 72.0% for direct LED-FM (P = 0.06) and direct ZN (P = 0.06), respectively. Specificity was 87.8% for LED-FM after NaOCl sedimentation compared to 96.7% and 95.9% for direct LED-FM (P<0.01) and direct ZN (P<0.01), respectively. Inter-reading agreement (kappa = 0.7) and technicians' acceptability were good.ConclusionNaOCl sedimentation did not improve the performance of LED-FM in the diagnosis of pulmonary TB at peripheral health service level.