Bloody Samples Research Articles

Utility of Acquire 22G-fine needle biopsy (FNB) needle vs the standard 22G needle during Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA): A Randomized Controlled Trial (RCT)

ABSTRACT Background Various types of needles are available to perform endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA). A relatively new needle for EBUS-TBNA, the Acquire Fine Needle Biopsy (FNB) device, has recently become available. Methods Consecutive subjects with lymphadenopathy >1 cm undergoing EBUS-TBNA were randomized to the Acquire 22-G EBUS-FNB needle and the standard 22-G EBUS-TBNA needle groups. Results A total of 86 subjects were randomized (43 in each group). The diagnostic yield of EBUS-TBNA was similar between the two groups: (36/43) 83.7% in the 22-G EBUS-FNB group and (34/43) 79.1% in the standard 22-G EBUS group (p = 0.58). The sampling adequacy, stations sampled, number of stations sampled, number of needle passes, and mean duration of the procedure between the two groups were also similar. Visible tissue clot core was obtained in a significantly greater proportion of subjects in the 22-G EBUS-FNB group (93.0% vs 46.5%, p < 0.001). Visibly bloody samples were more frequent in the 22-G EBUS-FNB group (74.4% vs 51.2%, p = 0.03). There was no difference in the complication rates between the two groups (p = 0.15). Conclusion We did not observe a difference in the diagnostic yield of the Acquire 22-G EBUS-FNB needle compared with the standard 22-G EBUS needle. Clinical trial registration Clinical Trial Registry of India (CTRI) https://ctri.nic.in/ (CTRI/2021/08/035589).

Thioflavin-modified molecularly imprinted hydrogel for fluorescent-based non-enzymatic glucose detection in wound exudate.

The concentration of glucose in the body's fluids is an important parameter that can indicate pathological conditions such as the progress of infected wounds. Several wearables and implantable detection approaches have been developed with high selectivity and sensitivity for glucose. However, all of them have drawbacks such as low stability, limited selectivity, and often require complex technology. In this work, we present a fluorescent-based cost-efficient imprinted hydrogel (MIH_GSH) capable of detecting glucose within 30 ​min. The imprinting approach allows us to improve the selectivity for glucose, overcoming the low specificity and limited binding efficiency at neutral pH of boronic acid-based detection mechanisms. The binding affinity determined for glucose-MIH_GSH was indeed 6-fold higher than the one determined for the non-imprinted hydrogel with a calculated imprinting factor of 1.7. The limit of detection of MIH_GSH for glucose in artificial wound exudate was calculated as 0.48 ​mM at pH 7.4 proving the suitability of the proposed approach to diagnose chronic wounds (ca. 1 ​mM). MIH_GSH was compared with a commercial colorimetric assay for the quantification of glucose in wound exudate specimens collected from hospitalized patients. The results obtained with the two methods were statistically similar confirming the robustness of our approach. Importantly, whereas with the colorimetric assay sample preparation was required to limit the interference of the sample background, the fluorescent signal of MIH_GSH was not affected even when used to measure glucose directly in bloody samples. The sensing mechanism here proposed can pave the way for the development of cost-efficient and wearable point-of-care tools capable of monitoring the glucose level in wound exudate enabling the quick assessment of chronic injuries.

The changing microRNA landscape by color and cloudiness: a cautionary tale for nipple aspirate fluid biomarker analysis

PurposeInvestigation of nipple aspirate fluid (NAF)-based microRNAs (miRNAs) as a potential screening tool for women at increased risk of developing breast cancer is the scope of our research. While aiming to identify discriminating NAF-miRNAs between women with different mammographic densities, we were confronted with an unexpected confounder: NAF sample appearance. Here we report and alert for the impact of NAF color and cloudiness on miRNA assessment.MethodsSeven classes of NAF colors coupled with cloudiness appearance were established. Using 173 NAF samples from 154 healthy women (19 samples were bilaterally collected), the expression of 14 target and 2 candidate endogenous control (EC) miRNAs was investigated using Taqman Advanced miRNA assays to identify significant differential expression patterns between color-cloudiness classes. Inter- and intra-individual variation of miRNA expression was analyzed using the coefficient of variation (CV).ResultsWe found that between the seven NAF classes, fold change miRNA expression differences ranged between 2.4 and 19.6 depending on the interrogated miRNA. Clear NAF samples exhibited higher miRNA expression levels compared to cloudy NAF samples with fold change differences ranging between 1.1 and 6.2. Inter-individual and intra-individual miRNA expression was fairly stable (CV < 15 %), but nevertheless impacted by NAF sample appearance. Within NAF classes, inter-individual variation was largest for green samples (CV 6-15 %) and smallest for bloody samples (CV 2-6 %).ConclusionsOur data indicate that NAF color and cloudiness influence miRNA expression and should, therefore, be systematically registered using an objective color classification system. Given that sample appearance is an inherent feature of NAF, these variables should be statistically controlled for in multivariate data analyses. This cautionary note and recommendations could be of value beyond the field of NAF-miRNAs, given that variability in sample color and cloudiness is likewise observed in liquid biopsies such as urine, cerebrospinal fluid and sputum, and could thereby influence the levels of miRNAs and other biomarkers.

New Cell Block Method to Enhance the Cellular Yield in Mucous and/or Bloody Samples

Objective: Cell blocks (CBs) are used to complement cytological diagnosis and for ancillary testing. Dissatisfaction with the cellular yield of the CB is widely recognized. Various techniques have been developed to increase the diagnostic utility of CBs. Study Design: We invented a new CB technique to increase cellular yield and diagnostic accuracy suitable especially for mucous and/or bloody cytological samples. Results: The new CB technique is described in detail with illustrations and cases, where it increased the cellular yield and diagnostic accuracy. CBs prepared by this method are suitable also for ancillary techniques, namely immunocytochemistry. Conclusions: The newly described method showed a better cellular yield in mucous and/or bloody cytological specimens.

Namlu-Hedef Arası Atış Mesafesi Tahmininde Tamamlayıcı Yardımcı Yöntem Olarak Kızılötesi Görüntüleme: Koyu, Desenli ve Kanlı Örnekler Üzerinde Bir Uygulama

Objective: The aim of this work was to assess the utility of the infrared camera as an effective tool for observing physical features like soot and gunshoot residues around the entrance hole, to aid estimation of the shooting distances on bloody, dark and patterned samples. Material and Methods: In this study, white control samples, as well as colored and patterned fabrics were fired from several distances (contact, 6 inches, 12 inches, 24 inches, and 36 inches). All shootings were performed with three replicates. Post-shooting infrared images were taken by use of Crime-lite 82S infrared, before and after application of blood on the samples. Human whole blood was sprayed onto the clothing by means of an aerosol spray bottle in two stages, and was partially dried before capturing the image with the infrared camera. Results: Using visual examination, it was not possible to detect any soot distribution on patterned fabrics, and the intensity of soot decreased and began to fade at 10 inches for other colored fabrics. The addition of blood to clothing masked the observation of iner soot and decreased the accuracy of the measurements. However, the soot was still visible when using an infrared camera on bloody samples, navy and black unknowns (10 inches) and patterned samples (contact and 6 inches), indicating the utility of an infrared camera on dirty and darker fabrics. Conclusion: The results of this study indicate that, similar to Modified Griess Test and Sodium Rhodizonate Test, soot alone is not the most accurate or reliable parameter for predicting true muzzle-to-target distance. However, an infrared camera enhanced the observation of presence of gunshoot residues not easily visible to the naked eye on dark, patterned and bloody samples. Thus, the proposed application of infrared imaging can easily be utilized as a complementary approach in the prediction of muzzle-to-target firing distance on dark, patterned and bloody fabrics.

Thyroid Fine-Needle Aspiration and Smearing Techniques.

Introduction: Thyroid fine-needle aspiration cytology is the most reliable preoperative diagnostic tool, but cases of failed or unsatisfactory diagnostic can occur. Therefore, we aim to improve aspiration and smearing techniques. We handle approximately 8000 thyroid fine-needle aspiration cytology cases annually. Here, we present the aspiration and smearing techniques resulting from our accumulated experience.Materials and Methods: Patients undergo aspiration cytology while seated on a barber chair, and are asked to gaze upwards to extend their anterior neck.1 Instead of relying on suction force, the samples are mainly obtained by cutting the tissue with needle movements. A strong negative pressure and a long aspiration time frequently produce bloody samples. Hence, we recommend negative pressure <0.3 mL and aspiration time up to 3 seconds. The obtained samples are placed on a glass slide and smeared using a second slide glass through a press and release method. When the samples are bloody, we tilt the glass slide, remove excess material, and wipe up the bloody components flowing from the slide. Liquid-based cytology is especially recommended for bloody or fluid samples. Biochemical measurement of thyroglobulin and calcitonin using fine-needle washout fluids is useful for diagnosing metastatic differentiated thyroid carcinoma and medullary thyroid carcinoma.2,3 When lymphoma is suspected, flow cytometry using aspirated samples is recommended.4Results: By applying the mentioned techniques and recommendations, we observed an increased accuracy in diagnosis and improved quality of our examinations.Conclusions: Fine-needle aspiration requires aspirating from the areas suitable for the diagnosis, obtaining adequate materials, and performing optimal smearing and fixation to retrieve highly accurate diagnoses. We hope our methods are helpful in improving your fine-needle aspiration cytology techniques, and result in more accurate cytological diagnoses. Thank you for taking interest in our methods.Acknowledgment: We would like to thank Dr. Louise Davies, Associate Professor of Surgery—Otolaryngology—Head and Neck Surgery, Geisel School of Medicine at Dartmouth, Hanover, NH, for her valuable comments on the creation of this video.We have no connection to any companies or products mentioned in this content. No competing financial interests exist.Runtime of video: 7 mins 15 secsThe Japanese version of this video was presented at the 29th Annual Congress of the Japan Association of Endocrine Surgeons, held in May 18, 2017, in Kobe, Japan.

Improved removal of blood contamination from ThinPrep cervical cytology samples for Raman spectroscopic analysis.

There is an unmet need for methods to help in the early detection of cervical precancer. Optical spectroscopy-based techniques, such as Raman spectroscopy, have shown great potential for diagnosis of different cancers, including cervical cancer. However, relatively few studies have been carried out on liquid-based cytology (LBC) pap test specimens and confounding factors, such as blood contamination, have been identified. Previous work reported a method to remove blood contamination before Raman spectroscopy by pretreatment of the slides with hydrogen peroxide. The aim of the present study was to extend this work to excessively bloody samples to see if these could be rendered suitable for Raman spectroscopy. LBC ThinPrep specimens were treated by adding hydrogen peroxide directly to the vial before slide preparation. Good quality Raman spectra were recorded from negative and high grade (HG) cytology samples with no blood contamination and with heavy blood contamination. Good classification between negative and HG cytology could be achieved for samples with no blood contamination (sensitivity 92%, specificity 93%) and heavy blood contamination (sensitivity 89%, specificity 88%) with poorer classification when samples were combined (sensitivity 82%, specificity 87%). This study demonstrates for the first time the improved potential of Raman spectroscopy for analysis of ThinPrep specimens regardless of blood contamination.

市販試薬を用いた高粘稠性検体および血性検体のための細胞診標本作製改良法

目的 : 良好な細胞診標本を作製することの難しい高粘稠性検体と血性検体の処理法を改善する.方法 : 遠心分離できない高粘稠性検体に検体量の 10%のサイトキープIIを添加し, 再遠心後に中間液層と沈渣のそれぞれをスライドグラス間ですり合わせて標本を作製した. 血性検体には沈渣に 5%リンス液加生理食塩水を約 10∼50 倍加え, 再遠心後, 標本を作製した. これらの方法で作製した標本と日本臨床細胞学会細胞検査士会 (JSCC) 法や未溶血法で作製した標本により診断した結果を比較した.成績 : 粘稠性検体 6 例をサイトキープ法で分析したところ, 全例で集細胞量が多く良好な染色性を示す標本を作製できた. その結果, JSCC 法で判定不能であった 5 例の判定が可能になった. 一方, 血性検体 30 例をリンス法で分析したところ, 未溶血法に比べ背景清浄, 細胞変性のない標本を短時間で作製できた. その結果 7 例で細胞診所見の改善が, 4 例で細胞診判定結果の改善がみられた.結論 : 新しい細胞診処理法により, 高粘稠性と血性検体の標本の質を改善することができた. 両方法とも迅速処理可能で細胞診の診断率向上に役立つと考える. 今後, 多数例を対象にした研究を計画し, 両法の有用性を確立したい.

Inferior Laryngeal Paraganglioma Mimicking A Primary Thyroid Tumor

To report an unusual case of inferior laryngeal paraganglioma that manifested as a thyroid neoplasm. A case report is presented, including ultra-sonographic, surgical, histologic, and immunohistochemical findings, and diagnostic and therapeutic strategies are discussed. In a 33-year-old man with a mass in the left side of his neck, color Doppler ultrasonography revealed an extremely hypervascular lesion that appeared to arise in the left lobe of the thyroid gland. Fine-needle aspiration was noncontributory because of bloody samples, but core biopsy specimens suggested a nonthyroidal neoplasm. At surgical intervention, a 3.5-cm mass was found immediately posterior to the left thyroid lobe, not involving the thyroid capsule. Because the mass could not be dissected free from the thyroid, performance of a left thyroid lobectomy and isthmectomy was necessary. The pathology specimen was a paraganglioma. Paragangliomas may be sonographically similar to thyroid neoplasms and may be included in the differential diagnosis of a hypervascular thyroid mass.

Acetic Acid Recovery of Gynecologic Liquid-Based Samples of Apparent Low Squamous Cellularity

To characterize cervicovaginal cytology samples with < 5,000 squamous cells on the initial ThinPrep slide (Cytyc Corp., Boxborough, Massachusetts, U.S.A) and to attempt sample recovery using acetic acid.Cervicovaginal cytology samples with <5,000 squamous cells on the original ThinPrep slide and residuum were reprocessed by adding 3 mL of 3:1 CytoLyt (Cytyc)/glacial acetic acid with production of a second slide. Both slides were reviewed for squamous cell quantitation and the presence of background material and abnormal cells.From a total of 1,833 cases, 147 (8.0%) were identified for reprocessing; 71 (48.3%) were grossly bloody and 58 (39.4%) grossly cloudy. Reprocessing resulted in a second slide with > 5,000 squamous cells in 116 (78.9%) cases and was most effective on cloudy samples (89.7% recovery) and bloody samples (71.8% recovery). Abnormal cells were identified in 13 (8.9%) reprocessed samples. In all but 2 cases the abnormal cells were present on the initial slide and demonstrated the same degree of abnormality as the reprocessed slide but were fewer in number.Acetic acid recovery increases squamous cell recovery when initially inadequate, reducing the number of unsatisfactory cases and in rare cases identifying a cytologically significant lesion not apparent on the original slide.

Impact of Needle Size on Pancreatic FNA Cytology

INTRODUCTION: 22-gauge needles are typically used for most pancreatic mass endoscopic ultrasound-guided fine-needle aspiration (EUS/FNA) procedures. It is unknown whether a larger or smaller gauge needle may actually provide better diagnostic material. 19-gauge needles may provide more diagnostic material but with more blood, while 25-gauge needles may provide less bloody samples but with less diagnostic material. The purpose of this preliminary study was to determine if there is a difference in cytologic yield, diagnostic material and background blood using various needle sizes for pancreatic FNA, and to determine whether an autopsy model could be used to test diagnostic yields for EUS/FNA. METHODS: The pancreas was removed during autopsies, which were performed within 24 hours of death. FNA was immediately performed upon removal using 1.5-inch long 19-, 22-, and 25-gauge needles attached to a 10cc aspiration syringe using negative pressure. Three passes were made with each needle. Aspirate smears were air-dried, alcohol-fixed, and stained with Diff-Quik and H&E stain, respectively. A single pathologist, blinded to the needle size used, evaluated each pass for cellularity and blood (1=low, 2=moderate, 3=high), and the presence of ductal cells, acinar cells and diagnostic material (yes/no), as defined by having an adequate number of cells for interpretation. RESULTS: Eleven pancreatic autopsy specimens were aspirated. Mean age of the patients was 51 years (range 38-77). None of the patients had known prior pancreatic disease. Results are shown below for the 24 passes obtained with each needle size. Diagnostic material was obtained from at least one pass for each needle size in all patients using the 25-guage needle, and in 7 of 8 using a 22- or a 19-gauge needles. CONCLUSIONS: 1) 19-, 22-, or 25-gauge needles for pancreatic FNA all provided adequate diagnostic material, with similar cellularity and background blood. 2) The use of an autopsy model for pancreatic FNA may be an effective means for comparing diagnostic yield using different EUS/FNA needles. 3) Larger needle size for EUS guided FNA of the pancreas may not improve diagnostic yield. Further research is needed to determine if needle size impacts cytologic yield in pancreatic malignancy and fibrosis.

Interphase-FISH-Test als Schnelltest für Trisomien im Fruchtwasser - Ergebnisse einer prospektiven Untersuchung*

Specific DNA probes allow rapid prenatal diagnosis of numerical chromosome disorders (chromosomes 13, 18, 21, X, Y) by FISH on interphase nuclei. The diagnostic reliability is presently under evaluation. In a period of 1.5 years a total 1126 amniotic fluid samples was investigated by FISH compared to standard cytogenetic analysis. The success rate was 93 percent (< or = 30 nuclei) and 84% (< or = 50 nuclei). An abnormal karyotype was detected by FISH in 27 of 28 successfully hybridised samples, including trisomy 21 [16], trisomy 13 [4], trisomy 18 [4], aberrations of sex chromosomes [4]. Two cases with clinically relevant cytogenetic abnormalities were in principle not detectable by FISH. One false-negative finding was observed, possibly arising from maternal cell contamination of the sample. 6% of all samples, respectively 23% of the bloody samples were contaminated by maternal cells (more than 10%). Maternal contamination represents the most important limitation of the diagnostic reliability in routine practice.

Risks for Fetal Abnormalities after very and moderately elevated AF-AFPs

In the 2 years between 1993 and 1995, we assayed alpha-fetoprotein (AFP) in 48,412 amniotic fluids (AFs). One thousand and eighty-six (2.2 per cent) measured > or = 2.0 MOM and these were subdivided into three groups; mildly (2.0-4.9 MOM), moderately (5.0-9.9 MOM), and very elevated (> or = 10.0 MOM). Abnormalities occurred in 25 per cent of the mildly elevated compared with 88 per cent of the moderately and 98 per cent of the very elevated cases. Forty-five per cent of the neural tube defects (NTDs) had AF-AFPs in the 5.0-9.9 and 36 per cent in the > or = 10.0 MOM range. After a positive acetylcholinesterase (AChE) test, both the moderately and the very elevated groups had abnormalities in > or = 95 per cent of cases, compared with 85 per cent in the mildly elevated group. After a negative AChE test, abnormalities occurred in 79, 52 and 18 per cent in the very elevated, moderately, and mildly elevated groups, respectively. Excluding chromosome abnormalities, an abnormal twin, and bloody samples, the risk of a fetal abnormality after a normal ultrasound was less than 1 per cent if the AChE was positive in both the moderately and the very elevated groups. If the AChE was negative, the risk was 18 per cent for the moderately and 55 per cent for the very elevated groups, of which congenital nephrosis accounted for 75 per cent; AFP levels usually increased in a later AF sample. Repeat amniocentesis may be offered to women with AF-AFPs > or = 5.0 MOM if no abnormality is seen with high-resolution ultrasound.

Maternal cell contamination in amniotic fluid samples as a consequence of the sampling technique

Maternal cell contamination in amniotic fluid samples is easily detected by in situ hybridization if the karyotype of the fetus differs from the karyotype of the mother. One out of two amniotic fluid samples appears to contain more than 20% maternal cells. Bloody samples often contain even more than 50% maternal cells. Maternal cells can also be identified on the basis of their nuclear morphology. Maternal cell contamination is regularly observed in pregnancies with an anterior placenta, whereas it is rare in posterior placenta pregnancies. The maternal cells are therefore thought to be artificially introduced into the amniotic fluid sample, as a result of placental bleeding during amniocentesis. The implications of maternal cell contamination for prenatal diagnosis using uncultured amniotic fluid samples are discussed.

Detection of errors in methylmalonyl-CoA metabolism by using amniotic fluid.

We report a method for rapid prenatal detection of methylmalonic acidemia, consisting of measuring methylmalonly-CoA mutase (EC 5.4.99.2) activity in non-cultured amniotic cells and measuring the concentration of methylmalonate in the amniotic fluid. Immediate stabilization of the mutase activity in the non-cultured amniotic cell by its coenzyme adenosycobalamin, and use of methylmalonyl-CoA with high specific activity gives mutase activity comparable to that of cultured amniotic cells or normal fibroblasts. Consequently, findings of low mutase activity and a hight concentration of methylmalonate in the amniotic fluid allows accurate diagnosis of the vitamin B12-nonresponsive form of methylmalonic acidemia. These results can be obtained in two days. For the vitamin B12-responsive form, the correct diagnosis depends upon finding amniotic fluid methylmalonate, because cells from these patients will display normal methylmalony-CoA mutase activity after adenosylcobalamin is added. Problems in interpreting data on bloody samples and the limitations of the method are discussed.