Cells In Blood Smears Research Articles

6985 Intravenous Immunoglobulin induced Type 1 diabetes remission

Abstract Disclosure: N. Shahid: None. D. Newbern: None. Introduction: Intravenous immunoglobulin [IVIG] is an immunomodulatory agent used for treatment of autoimmune and inflammatory conditions. However, it is not indicated in patients with type 1 diabetes [T1D] as its efficacy has not been established. Clinical Case: A 9-year-old previously healthy boy presented to the Emergency Department with several days of unexplained bruising and mucosal bleeding with no prior history of trauma, bleeding disorders or other systemic symptoms. Physical examination showed generalized petechiae, ecchymosis and purpura. Vitals were normal including weight and BMI. Initial labs confirmed Immune thrombocytopenic purpura [ITP] as his platelets were less than 1K/µL with otherwise normal peripheral blood cell counts and smear. Concomitantly, labs showed significant hyperglycemia [260mg/dL] with glycosuria and trace ketonuria. Glycated hemoglobin [Hba1c] was elevated at 10.5% and ultimately Pancreatic autoantibodies [GAD65, Zinc transporter-8 and islet cell antibody] came back positive confirming T1D. Other autoimmune disorders such as adrenal insufficiency, thyroiditis, and celiac disease were ruled out. For the treatment of ITP, he the patient received 1 dose of IVIG 1g/kg and responded well with a rise in platelets to 6K/µL. The morning following the infusion, due to fasting hyperglycemia, the patient was started on basal-bolus insulin regimen. By 2nd day post IVIG, he had hypoglycemia, so insulin was discontinued with normalization of blood sugars. He is now 7 months post IVIG with no recurrent episodes of thrombocytopenia. He also remains off insulin with Hba1c in the 5.9-6.5% range. He eats a regular diet with avoidance of simple carbohydrates. He wears onwears a a continuous glucose monitor, which shows that blood sugars are in range 93% of the time He remains euglycemic overnight with transient self-resolving mild post-prandial hyperglycemia. Pancreatic autoantibodies have been reassessed and remain positive Conclusions: This case demonstrates that IVIG used for the treatment of ITP has induced a state of remission in our patient with newly diagnosed with T1D. IVIG is not a standard of treatment for T1D, however, when given for other autoantibody- or T-cell-mediated autoimmune conditions, it may reduce the insulin need in patients with known diabetes or induce remission in patients with newly diagnosed diabetes.

Efficient deep learning-based approach for malaria detection using red blood cell smears

Malaria is an extremely malignant disease and is caused by the bites of infected female mosquitoes. This disease is not only infectious among humans, but among animals as well. Malaria causes mild symptoms like fever, headache, sweating and vomiting, and muscle discomfort; severe symptoms include coma, seizures, and kidney failure. The timely identification of malaria parasites is a challenging and chaotic endeavor for health staff. An expert technician examines the schematic blood smears of infected red blood cells through a microscope. The conventional methods for identifying malaria are not efficient. Machine learning approaches are effective for simple classification challenges but not for complex tasks. Furthermore, machine learning involves rigorous feature engineering to train the model and detect patterns in the features. On the other hand, deep learning works well with complex tasks and automatically extracts low and high-level features from the images to detect disease. In this paper, EfficientNet, a deep learning-based approach for detecting Malaria, is proposed that uses red blood cell images. Experiments are carried out and performance comparison is made with pre-trained deep learning models. In addition, k-fold cross-validation is also used to substantiate the results of the proposed approach. Experiments show that the proposed approach is 97.57% accurate in detecting Malaria from red blood cell images and can be beneficial practically for medical healthcare staff.

Forms of degeneration of blood cells, their physiological and clinical significance, mechanisms of formation, shadows of cells in blood smears of birds

Relevance. An overview is presented of the patterns of appearance, physiological and clinical significance of shadows of cells, a comparative experimental cytomorphological analysis using the example of polychromatophilic erythroblasts (PolyErythro), shadows of erythroblast (ShadErythro) and undifferentiated shadows cells of in peripheral blood smears of chickens Gallus gallus L.Methods. The studied individuals were divided into four groups (n = 40) depending on the age of the chickens (Postembryonalis — P1, P7, P23, and P42): 1, 7, 23, and 42 days of postembryonic ontogenesis) of a factory herd. ShadErythro apoptotic changes, undifferentiated cell shadows were characterized. Were calculated and characterized the morphophysiological parameters of PolyErythro, ShadErythro and unidentified shadows of cells by high-resolution color microphotographs in blood smears of birds (Pappenheim stained); a total of 158 (n = 158) microphotographs were analyzed.Results. The structured ShadErythro were able in chromatinolized. Unidentified shadows of cells were in a state of cytolysis, fragmentation of the cytoplasm, karyopycnosis and karyorrhexis, chromatin fragmentation, and karyolysis were noted. Structured of ShadErythro and undifferentiated shadows of cells were light purple with a reddish tint. Significant difference in the area of ShadErythro (Sshadow, μm2) was recorded in the ascending area of PolyErythro (Scell, μm2), within 25% (p ≤ 0.001), an increase in the percentage of difference between the minimum and maximum values of the ShadErythro area (Pshad.-cell-differ.-min-max,%) of Pcell-differ.-min-max,% PolyErythro was 36% (p ≤ 0.001). The difference in the percentage difference between the minimum and maximum values of the area of the nucleus of erythroblasts (Pnucleus-differ.-min-max) and the shadows of the nucleus of erythroblasts (Pshad.-nucle.-differ.-min-max) is significantly less than the analogous values (noted above) for cells and shadows erythroblasts, for example, Pnucleus-differ.-min-max exceeded Pshad.-nucle.-differ.-min-max by only 21% (p ≤ 0.001). The percentage of difference between the minimum and maximum values of the nuclear-cytoplasmic ratio ShadErythro (PN/C Ratio-shad.-cell) significantly exceeded the analogous value of PolyErythro (PN/C Ratio) within 64% (p ≤ 0.001). At the same time, the nuclear-cytoplasmic ratio of PolyErythro (N/C Ratio) and ShadErythro (N/C Ratio-shad.-cell) had a relatively minimal difference. The ShadErythro nuclear-cytoplasmic ratio (N/C Ratio-shad.-cell) exceeded the PolyErythro nuclear-cytoplasmic ratio (N/C Ratio) by only 12% (p ≤ 0.001). ShadErythro is maximally represented in chickens aged P1 — 40% and up to 30% (p ≤ 0.05) in P23. In P7, the amount of ShadErythro decreased by 30% (p ≤ 0.05), in P42, a decrease in ShadErythro to 20% (p ≤ 0.05) was recorded. The described dynamics of ShadErythro is due to physiological adaptive processes in the body of broiler chickens in the early period of postembryonic ontogenesis. The data obtained by us from a comprehensive comparative morphophysiological analysis of forms of degeneration of blood cells will be useful in the clinical diagnosis of animal pathologies, experimental and theoretical studies of cellular adaptations of the functional blood system and the whole organism.

Automated Detection of Malaria and Sickle Cells in Blood Smears Using Edge Detection and Morphological Operations

Automated Detection of Malaria and Sickle Cells in Blood Smears Using Edge Detection and Morphological Operations

Malaria parasite detection using deep learning algorithms based on (CNNs) technique

Malaria is a life-threatening disease caused by female anopheles mosquito bites that are prevalent in many regions of the world. We introduce a deep convolutional neural network (CNN) to improve malaria diagnosis accuracy using patches segmented from microscopic images of red blood cell smears. We design the automatic parasite detection in blood from Giemsa-stained smears using three CNN pre-trained models such as VGG19, ResNet50, and MobileNetV2. As the CNNs are poorly performing for small datasets, we introduce the transfer learning technique. Transfer learning involves acquiring visual features from large general datasets and resolving issues using small datasets. We use a transfer learning approach to detect and classify malaria parasites with three CNN pre-trained models. We evaluated proposed CNN models experimentally using the National Institute of Health (NIH) Malaria Dataset. Our proposed model achieves an accuracy of almost 100%.

Single Channel Image Enhancement (SCIE) of White Blood Cells Based on Virtual Hexagonal Filter (VHF) Designed over Square Trellis.

White blood cells (WBCs) are the important constituent of a blood cell. These blood cells are responsible for defending the body against infections. Abnormalities identified in WBC smears lead to the diagnosis of disease types such as leukocytosis, hepatitis, and immune system disorders. Digital image analysis for infection detection at an early stage can help fast and precise diagnosis, as compared to manual inspection. Sometimes, acquired blood cell smear images from an L2-type microscope are of very low quality. The manual handling, haziness, and dark areas of the image become problematic for an efficient and accurate diagnosis. Therefore, WBC image enhancement needs attention for an effective diagnosis of the disease. This paper proposed a novel virtual hexagonal trellis (VHT)-based image filtering method for WBC image enhancement and contrast adjustment. In this method, a filter named the virtual hexagonal filter (VHF), of size 3 × 3, and based on a hexagonal structure, is formulated by using the concept of the interpolation of real and square grid pixels. This filter is convolved with WBC ALL-IBD images for enhancement and contrast adjustment. The proposed filter improves the results both visually and statically. A comparison with existing image enhancement approaches proves the validity of the proposed work.

Leukemoid reaction in a patient with COVID-19 infection

Rationale: COVID-19 usually presents with flu-like symptoms and signs, but some rare presentations like leukemoid symptoms cannot be ignored. Patient's concerns: A 37-year-old female presented to the outpatient department with flu-like symptoms. Diagnosis: The RT-PCR test for SARS-CoV-2 infection was positive, while complete blood cell and peripheral blood smear showed leukemoid reaction. Intervention: Paracetamol and fexofenadine for flu-like symptoms and leukemoid presentation. Outcomes: On the 10th day, the patient was asymptomatic and RT-PCR was negative. So the patient was discharged and leukemoid presentation subsided after clearance of viral disease. Lessons: COVID-19 has a myriad of presentations, and unusual symptoms/signs especially in this pandemic could be induced by COVID-19 infection.

Comparative staining of Rhinolophus spp. white blood cells in blood smears.

Background and Aim:A drawback of studies on bat blood smears in the field is the lack of time for fixation because blood sampling using a non-lethal method often provides less time for fixation in smear preparations due to the small volume of blood collected. Usually, there is insufficient blood for another smear preparation, so it is necessary to use blood smears as rationally as possible, especially for rare bats. Many stains are used for staining peripheral blood smears, and they have advantages and disadvantages. This study aimed to examine commonly used stains for blood smears to select the best stain for staining peripheral blood smears in bats.Materials and Methods:In this study, 48 blood smears of Rhinolophus spp. bats were examined using several blood staining methods. Four methods that showed the best results were used in further experiments: Romanowsky-Giemsa, Pappenheim, hematoxylin-eosin, and eosin methylene blue.Results:Comparative analysis of different methods for staining bat blood smears revealed that the most convenient method for analyzing blood cells is Pappenheim method.Conclusion:Staining blood smears using Pappenheim method yield the least number of unsuccessful blood smear stains and are quite effective for the morphological analysis of blood cells.

Dietary alpha‐linolenic acid reduces platelet activation and collagen‐mediated cell adhesion in sickle cell disease mice

BackgroundSickle cell disease (SCD) is a genetic hemoglobinopathy associated with high morbidity and mortality. The primary cause of hospitalization in SCD is vaso‐occlusive crisis (VOC), mediated by alteration of red blood cells, platelets, immune cells and a pro‐adhesive endothelium. ObjectivesWe investigated the potential therapeutic use of the plant‐derived omega‐3 alpha‐linolenic acid (ALA) in SCD. MethodsBerkeley mice were fed a low‐ or high‐ALA diet for 4 weeks, followed by analysis of liver fibrosis, endothelial activation, platelet activation and formation of platelet‐neutrophils aggregates. Aggregation of platelets over collagen under flow after high‐ALA was compared to a blocking P‐selectin Fab. ResultsDietary high‐ALA was able to reduce the number of sickle cells in blood smear, liver fibrosis, and the expression of adhesion molecules on the endothelium of aorta, lungs, liver and kidneys (VCAM‐1, ICAM‐1 and vWF). Specific parameters of platelet activation were blunted after high‐ALA feeding, notably P‐selectin exposure and the formation of neutrophil‐platelet aggregates, along with a correspondingly reduced expression of PSGL‐1 on neutrophils. By comparison, in vivo treatment of SCD mice with the anti‐P‐selectin Fab was able to similarly reduce the formation of neutrophil‐platelet aggregates, but did not reduce GpIbα shedding nor the activation of the αIIbβ3 integrin in response to thrombin. Both ALA feeding and P‐selectin blocking significantly reduced collagen‐mediated cell adhesion under flow. ConclusionsDietary ALA is able to reduce the pro‐inflammatory and pro‐thrombotic state occurring in the SCD mouse model and may represent a novel, inexpensive and readily available therapeutic strategy for SCD.

WBC-AMNet: Automatic classification of WBC images using deep feature fusion network based on focalized attention mechanism.

The recognition and classification of White Blood Cell (WBC) play a remarkable role in blood-related diseases (i.e., leukemia, infections) diagnosis. For the highly similar morphology of different WBC subtypes, it is too confused to classify the WBC effectively and accurately for visual observation of blood cell smears. This paper proposes a Deep Convolutional Neural Network (DCNN) with feature fusion strategies, named WBC-AMNet, for automatically classifying WBC subtypes based on focalized attention mechanism. To obtain more localized attention of CNN, the fusion features of the first and the last convolutional layer are extracted by focalized attention mechanism combining Squeeze-and-Excitation (SE) and Gather-Excite (GE) modules. The new method performs successfully in classifying monocytes, neutrophils, lymphocytes, and eosinophils on the complex background with an overall accuracy of 95.66%, better than that of general CNNs. The multi-classification accuracy of WBC-AMNet with the background segmentation is over 98% in all cases. In addition, Gradient-weighted Class Activation Mapping (Grad-CAM) is employed to visualize the attention heatmaps of different feature maps.

Jaundice in a Preterm Infant Twelve Hours After Birth.

Jaundice in a Preterm Infant Twelve Hours After Birth.

On the Effectiveness of Leukocytes Classification Methods in a Real Application Scenario

Automating the analysis of digital microscopic images to identify the cell sub-types or the presence of illness has assumed a great importance since it aids the laborious manual process of review and diagnosis. In this paper, we have focused on the analysis of white blood cells. They are the body’s main defence against infections and diseases and, therefore, their reliable classification is very important. Current systems for leukocyte analysis are mainly dedicated to: counting, sub-types classification, disease detection or classification. Although these tasks seem very different, they share many steps in the analysis process, especially those dedicated to the detection of cells in blood smears. A very accurate detection step gives accurate results in the classification of white blood cells. Conversely, when detection is not accurate, it can adversely affect classification performance. However, it is very common in real-world applications that work on inaccurate or non-accurate regions. Many problems can affect detection results. They can be related to the quality of the blood smear images, e.g., colour and lighting conditions, absence of standards, or even density and presence of overlapping cells. To this end, we performed an in-depth investigation of the above scenario, simulating the regions produced by detection-based systems. We exploit various image descriptors combined with different classifiers, including CNNs, in order to evaluate which is the most suitable in such a scenario, when performing two different tasks: Classification of WBC subtypes and Leukaemia detection. Experimental results have shown that Convolutional Neural Networks are very robust in such a scenario, outperforming common machine learning techniques combined with hand-crafted descriptors. However, when exploiting appropriate images for model training, even simpler approaches can lead to accurate results in both tasks.

Effect of supplementation with dried fruit pomace on the performance, egg quality, white blood cells, and lymphatic organs in laying hens

The aim of the current study was to assess the impact of raspberry (RA), black currant (BC), and black chokeberry (CA) dried pomace, at 30 g per kg of feed, on performance, egg quality, white blood cells, heterophil:lymphocyte ratio, and lymphatic organs in Hy-Line laying hens. Hens fed a basal diet comprised the control group (CO). A total of 480, 42-week-old Hy-Line commercial hybrid laying hens were randomly divided into 4 equal groups differing in the diet: in control group (CO), the birds received a standard diet, in groups CH, BC and RA, the birds were fed with a standard diet enriched with 30 g/kg of dried fruit pomace: Black chokeberry, black currant, and raspberry, respectively.Differences in yolk color were observed between the CO and groups BC and RA, as well as between groups CA and BC (P < 0.001). The eggs of group RA were characterized by a lower weight (P = 0.001 and P = 0.002) and shell density (P = 0.025 and P = 0.002) in comparison to eggs from the control group CO. The lightest yolk color was observed in the eggs of group BC in comparison with the other groups (P = 0.006). The greatest yolk weight was observed for group CH, while the smallest was for group CO (P = 0.017). Laying hens in the groups BC and RA were characterized at wk 62 of age by a higher percentage of spleen in comparison with the control group CO (P = 0.018).In conclusion, the BC and RA diet resulted in a paler yolk color and was associated with slightly poorer shell quality parameters. On the other hand, the CH and BC diet had a positive impact on the immune system of the laying hens, as confirmed by the changes in the white blood cell smear and the higher spleen percentage. Therefore, the use of these by-products in the diets of poultry, i.e., dried pomace of black chokeberry and black currant may have a positive impact by improving the immunological status of laying hens.

Dynamics of morphological, immunological and histological changes in microsporіа in guinea pigs

Microsporіа affect different species of animals and humans. The high contagiousness of the pathogen determines the relevance of research into this disease. Microsporum canis is the pathogen that most often causes microsporia. Weakened functions of the immune system and violation of the epithelial barrier of the skin are a favourable factor that causes microspores. The main source of infection is cats, which are involved in the storage and transmission of the pathogen. To clarify the dynamics of morphological, immunological and histological changes in microsporia, blood and skin studies of guinea pigs infected with M. canis were carried out. The animals were divided into two groups of 6 guinea pigs (healthy and sick). Test material (blood and skin) was taken from clinically healthy and sick animals 21 and 42 days after infection. The number of erythrocytes and leukocytes was determined by counting them in the Goryaev chamber, the hemoglobin content – by the method of cyanide hemoglobin. The leukogram was derived based on the counting and differentiation of 200 leukocyte cells in blood smears. Material for histological examination (pieces of skin) was fixed in 10–12% cooled solution of neutral formalin, followed by pouring in paraffin according to the scheme proposed by G. A. Merkulov. The obtained results demonstrated that leukocytosis developed in guinea pigs with microsporia on the 21st and 42nd days; the number of rod-shaped neutrophils increased, that of segmental neutrophils decreased, and that of ESR increased. The immune response to the course of microsporia was manifested in an increase in the percentage of T-lymphocytes, T-suppressors and a decrease in T-helper cells and an increase in T-killers compared with healthy animals. Histological examination showed that on the 21st day after infection, hyphae and spores of the fungus M. canis were localized in the skin. There is swelling of the dermis, stratification of collagen fibers and the accumulation of inflammatory infiltrates around the hair follicles. On the 42nd day, the infiltration spread and dystrophic changes in the skin occurred in the form of desquamation of the epidermis and the formation of acanthosis and hyperkeratosis on the surface of the dermis. The conducted research will allow further assessment of the course of microsporia under the action of various drugs and help establish the most effective method of treatment.

Novel defocus hologram aberration compensation method in digital holography integrated pathology microscope for label free 3-D imaging

In this paper we report successful integration of digital holography in off- axis transmission configuration into a commercially available pathology microscope (PathologyDHM), where Mach-Zehnder interferometric configuration was achieved in pathology microscope using 1 × 2 single mode fiber coupler and custom built optical modules. A smartphone/webcam positioned at the image plane of the finite conjugate microscope captures the interference pattern generated by the system. Using PathologyDHM, we have demonstrated a novel and simple method for compensation of aberration due to microscope objective (MO). The proposed method is called defocus hologram method and it is particularly useful in cases, where samples do not have specimen free or background flat region. Proper validation of the method was done by imaging a transmissive grating (commercial compact disc, CD-R), which doesn't have a flat background region and comparing phase profile of CD-R computed by our proposed method with self-reference conjugated hologram method. For reiterating its biological applications, phase profile of a biological sample, red blood cell smear (RBC smear), was computed with our defocus hologram method and compared with already established reference conjugated hologram method. Phase profiles of CD-R and RBC smear computed with defocus hologram method agree with self-reference and reference conjugated hologram methods respectively, within the phase error limits of the system. Integration of digital holography in to commercial pathology microscope enables 3D imaging, which coupled with artificial intelligence may prove useful in disease diagnostics and monitoring response of cells to treatment.

K-Means Clustering Based on Otsu Thresholding For Nucleus of White Blood Cells Segmentation

White blood cells function as the human immune system, and help defend the body against viruses. In clinical practice, identification and counting of white blood cells in blood smears is often used to diagnose many diseases such as infection, inflammation, malignancy, leukemia. In the past, examination of blood smears was very complex, manual tasks were tedious and time-consuming. This research proposes the k-means clustering algorithm to separate white blood cells from other parts. However, k-means clustering has a weakness that is when determining the initial prototype values, so the otsu thresholding method is used to determine the threshold by utilizing global values, then proceed with morphological operations to refine the segmentation image. The results of segmentation are measured by the Positive Predeictive Value (PPV) and Negative Positive Value (NPV) parameters. The results obtained prove that the use of otsu thresholding and morphological operations significantly increase the value of PPV compared to the value of PPV that does not use otsu thresholding. Whereas the NPV value increased but not significantly.

Automated screening of sickle cells using a smartphone-based microscope and deep learning

Sickle cell disease (SCD) is a major public health priority throughout much of the world, affecting millions of people. In many regions, particularly those in resource-limited settings, SCD is not consistently diagnosed. In Africa, where the majority of SCD patients reside, more than 50% of the 0.2–0.3 million children born with SCD each year will die from it; many of these deaths are in fact preventable with correct diagnosis and treatment. Here, we present a deep learning framework which can perform automatic screening of sickle cells in blood smears using a smartphone microscope. This framework uses two distinct, complementary deep neural networks. The first neural network enhances and standardizes the blood smear images captured by the smartphone microscope, spatially and spectrally matching the image quality of a laboratory-grade benchtop microscope. The second network acts on the output of the first image enhancement neural network and is used to perform the semantic segmentation between healthy and sickle cells within a blood smear. These segmented images are then used to rapidly determine the SCD diagnosis per patient. We blindly tested this mobile sickle cell detection method using blood smears from 96 unique patients (including 32 SCD patients) that were imaged by our smartphone microscope, and achieved ~98% accuracy, with an area-under-the-curve of 0.998. With its high accuracy, this mobile and cost-effective method has the potential to be used as a screening tool for SCD and other blood cell disorders in resource-limited settings.

Epimedium Polysaccharide Ameliorates Benzene-Induced Aplastic Anemia in Mice.

Benzene (BZ) is an important occupational and environmental pollutant. Exposure to BZ may cause aplastic anemia which is characterized as bone marrow hematopoietic failure. In order to reduce the harmful effects of this pollutant, it is necessary to identify additional preventative measures. In this study, we investigated the protective effects of epimedium polysaccharide (EPS), a natural compound with antioxidant and immune-enhancing potency, on aplastic anemia induced by benzene exposure in mice. Male CD-1 mice were randomly divided into five groups including control, BZ (880 mg/kg), LE (EPS low-dose, 20 mg/kg + BZ), ME (EPS middle-dose, 100 mg/kg + BZ), and HE (EPS high-dose, 200 mg/kg + BZ) groups. Animals were exposed to BZ by subcutaneous injection in the presence or absence of EPS via oral administration. All mice were treated 3 times a week for 8 consecutive weeks to develop a mouse model of benzene-induced aplastic anemia (BIAA). Results showed that BZ induced a significant decrease in both white and red blood cells, platelet counts, and hemoglobin level compared with that in the control group (p < 0.01). Treatment of EPS led to a protective effect against these changes particularly in the highest-dose group (HE, p < 0.01). EPS also recovered the decreased number of nucleated cells in peripheral blood cell smears and femur biopsies by BZ exposure. The increased level of reactive oxygen species (ROS) in bone marrow mononuclear cells (BMMNCs) in mice from the BZ group was significantly lower (p < 0.01) in the mice from the highest concentration of EPS (HE) group when compared with that from the control group. In addition, BZ exposure led to a significant increase in the apoptosis rate in BMMNCs which was prevented by EPS in a dose-dependent manner (p < 0.01). The antiapoptosis effect of EPS was through reversing apoptotic proteins such as BAX, Caspase-9 and Caspase-3, and Bcl-2. Finally, EPS treatment partially restored the levels of T cells and the different subtypes except CD80+ and CD86+ compared with the BZ group (HE, p < 0.05). These results suggest that EPS has protective effects against BIAA via antioxidative stress, immune modulation, and antiapoptosis mechanisms.

White Blood Cell Profiles in Long-Term Captive and Recently Captured Eastern Tiger Salamanders (Ambystoma tigrinum)

Physiological responses to stress are important indicators of the effects of environmental disruption on individuals and, therefore, a way to determine the health of populations. These responses can be measured in a variety of ways, including the survey of differential counts of white blood cells. Baseline cell numbers and neutrophil to lymphocyte ratios have been determined for a number of species in the genus Ambystoma, but not for A. tigrinum, and baseline values are necessary for assessments of stress in natural and captive populations. We counted white blood cells in blood smears from long-term captive and recently captured Eastern Tiger Salamanders (A. tigrinum) and compared the proportions of each cell type between these two samples. We also compared our results to the published values for other post-metamorphic or paedomorphic ambystomatids. Mean neutrophil to lymphocyte ratios, a measure of stress, were higher in our captive salamanders (0.81) than in the wild sample (0.41), as were the mean number of basophils (36.0 for captive and 10.3 for wild). The cell counts for our wild salamanders were comparable to those for other unstressed ambystomatids. Our results suggest that our long-term captive salamanders are under a small degree of stress and are not a good source of baseline values for this species.

EXPANDING THE ROLE OF MAGNETIC FIELDS IN RED BLOOD CELLS DEFORMATIONS: DEMONSTRATION OF PARAMAGNETIC AND DIAMAGNETIC FIELDS

Background: The main purpose of this manuscript is to introduce a biophysics based mechanism of the role of magnetism in temporary cell deformation. Biomagnetism: Biomagnetism has been slowly introduced in the medical literature as follows: The diamagnetic nature of blood reported in1936, the recording of organ emissions from the human body also published in the second half of the last century. It could stated that the same way molecular cross talk influence cells signaling, biomagnetic cross talk could also influence cells functions. Introduction: This manuscript introduces the effect of externally applied magnetic fields (diamagnetic or paramagnetic) causing temporary Red Blood Cells (RBCs) deformations. Results from experiments using diamagnetic material such as human blood with the property of repelling an opposing magnetic field; and paramagnetic material such as carbon based graphite or iron particles are reported. Methods: The work herein presented entailed the trapping (between two similar glass slides) of fresh human blood smear with fine exogenous metal, such as iron filings or graphite powder. The technique requires a standard video microscope and an image recording equipment. Provocative placement of ferromagnetic or diamagnetic material in in vitro blood smears preparations were found to exhibit temporary in vivo characteristics quoted Temporary In Vivo Blood Smears (TBIS). Established protocol calls for fresh blood smears to be set-aside 5 minutes prior to staining. During the first 2 minutes of the set-aside period, in a typical monolayer smear, one can observe an initial brief time period of approximately 1’ ± 30” of a drying cycle. Usually, a moisture sheen boundary is seen gradually moving from the slide’s edges towards the center. Results: The main findings include a demonstration that both diamagnetic and pararmagnetic material deform RBCs in a variety of shapes. The paramagnetism (attraction) and diamagnetism of graphite powder induced temporary RBCs deformation in the form of teardrops. For the first time, an ongoing cellular deformation process was captured in video recordings. Conclusions: When in vivo cells in blood smears, such as RBCs, are fronted by exogenous magnetic fields, temporary cell deformation occurs.