-
The Journal of Experimental Medicine May 1953The cells of a human epithelial cancer cultivated en masse have been shown to support the multiplication of all three types of poliomyelitis virus. These cells (strain...
Studies on the propagation in vitro of poliomyelitis viruses. IV. Viral multiplication in a stable strain of human malignant epithelial cells (strain HeLa) derived from an epidermoid carcinoma of the cervix.
The cells of a human epithelial cancer cultivated en masse have been shown to support the multiplication of all three types of poliomyelitis virus. These cells (strain HeLa of Gey) have been maintained in vitro since their derivation from an epidermoid carcinoma of the cervix in February, 1951. As the virus multiplied it caused in from 12 to 96 hours degeneration and destruction of the cancer cells. The specific destructive effect of the virus was prevented by adding homotypic antibody to the cultures but not by adding heterotypic antibodies. Methods for the preparation of large numbers of replicate cultures with suspensions of strain HeLa cells were described. The cells in suspension were readily quantitated by direct counts in a hemocytometer. A synthetic solution that maintains cellular viability was employed for viral propagation. The experimental results demonstrate the usefulness of strain HeLa cells for (a) the quantitation of poliomyelitis virus, (b) the measurement of poliomyelitis antibodies, and (c) the production of virus.
Topics: Antibodies; Antibodies, Heterophile; Carcinoma, Squamous Cell; Cervix Uteri; Female; HeLa Cells; Humans; In Vitro Techniques; Poliomyelitis; Poliovirus; Tissue Culture Techniques; Viruses
PubMed: 13052828
DOI: 10.1084/jem.97.5.695 -
Acta Clinica Croatica Sep 2020Recently, studies have reported that inflammatory response and elevated platelet counts are associated with several cancers. In the present study, we aimed to evaluate...
Recently, studies have reported that inflammatory response and elevated platelet counts are associated with several cancers. In the present study, we aimed to evaluate hemocytometer parameters in differentiating adrenal adenoma and carcinoma, and the prognostic utility of hemocytometer parameters in adrenocortical carcinoma (ACC). We included 30 patients with nonfunctional adrenal adenoma and 13 patients with ACC having undergone surgery between 2005 and 2017 and followed up postoperatively at our centre. The neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR), red blood cell distribution width (RDW), mean platelet volume (MPV) and plateletcrit (PCT) were evaluated preoperatively in all patients included in the study. There was a significant difference between the adrenal adenoma and ACC groups in terms of neutrophil and lymphocyte counts, NLR and PLR. There was no significant difference between the two groups in terms of platelet count and MPV, but PCT levels were significantly lower in ACC group. There was no statistically significant difference between recurrent and/or metastasis positive patients and negative ones according to NLR, PLR, RDW and MPV. There was a statistically significant difference in RDW levels and tumor diameter between the groups. Our study is the first to evaluate hemocytometer parameters in differentiating adrenal adenomas and carcinomas, and also in the prognosis of ACC. The present study suggested that the hemocytometer parameters may be a marker in the differential diagnosis of adrenal adenomas and carcinomas. However, our study also showed that these parameters had no prognostic value in ACC.
Topics: Adenoma; Adrenal Cortex Neoplasms; Adrenocortical Carcinoma; Biomarkers; Humans; Lymphocytes; Neutrophils; Platelet Count; Prognosis
PubMed: 34177053
DOI: 10.20471/acc.2020.59.03.07 -
Theriogenology Jun 2016This article is the result of the work of the andrology task-force of the Association of Applied Animal Andrology, American College of Theriogenologists, European... (Review)
Review
This article is the result of the work of the andrology task-force of the Association of Applied Animal Andrology, American College of Theriogenologists, European College of Animal Reproduction, Society for Theriogenology, and National Association of Animal Breeders. It is intended to serve as a comprehensive reference on methods to evaluate sperm concentration and to contribute to the adoption of best practices in veterinary andrology laboratories. The information covered in the article includes sample preparation and the use of manual counts, spectrophotometers, computer-assisted semen analysis, NucleoCounter, and flow cytometry. Emphasis is given to the principles of the methods and equipment, performing the evaluation, and common mistakes and/or pitfalls. In addition, the precision and accuracy of the different methods are also discussed.
Topics: Flow Cytometry; Image Processing, Computer-Assisted; Practice Guidelines as Topic; Semen Analysis; Species Specificity; Specimen Handling; Spectrophotometry; Sperm Count
PubMed: 27045626
DOI: 10.1016/j.theriogenology.2016.01.002 -
JBRA Assisted Reproduction Mar 2024The Neubauer hemocytometer, as well as the Makler chamber, are devices commonly used in andrology laboratories. The present study aimed to verify if both methods yield...
OBJECTIVE
The Neubauer hemocytometer, as well as the Makler chamber, are devices commonly used in andrology laboratories. The present study aimed to verify if both methods yield comparable results, and whether they can be used interchangeably to determine sperm concentration.
METHODS
Sperm and latex beads concentration measurements were performed with the Neubauer hemocytometer and the Makler chamber. Fixed and proportional biases were estimated, and the method agreement was determined by assessing sperm concentration results with the Bland and Altman plot. The Coefficient of Variation (CV) and relative bias were calculated as an index of precision and accuracy, respectively, by measuring latex beads target concentrations in both chambers.
RESULTS
The Makler chamber systematically overestimated the Neubauer hemocytometer concentration measurements by a mean of -7.99%, with limits of agreement (LOA) between -41% to 25.61% (p<0.001). The fixed bias was found for concentration values inferior to 40 x 106/ml range (p<0.001), but not higher concentration results (p>0.05). Measurements with the Neubauer hemocytometer showed the greatest consistency in the study with the CV ranging from 3.01% to 6.67%; while the CV with the Makler chamber ranged from 8.46% to 25.64%. The relative bias for the Neubauer hemocytometer determinations varied from 0.12% to 8.40%, while for the Makler chamber varied from 7.6% to an overestimation of 38.0%.
CONCLUSIONS
Measurements made with the Makler chamber demonstrated more variability and a higher degree of overestimation. The Makler chamber is a poor substitute to the Neubauer hemocytometer for evaluation of oligozoospermic samples, although both chambers render similar results for highly concentrated samples.
PubMed: 38530757
DOI: 10.5935/1518-0557.20240023 -
Advances in Laboratory Medicine May 2021Body fluid cell counting provides valuable information for the diagnosis and treatment of a variety of conditions. Chamber cell count and cellularity analysis by optical... (Review)
Review
Body fluid cell counting provides valuable information for the diagnosis and treatment of a variety of conditions. Chamber cell count and cellularity analysis by optical microscopy are considered the gold-standard method for cell counting. However, this method has a long turnaround time and limited reproducibility, and requires highly-trained personnel. In the recent decades, specific modes have been developed for the analysis of body fluids. These modes, which perform automated cell counting, are incorporated into hemocytometers and urine analyzers. These innovations have been rapidly incorporated into routine laboratory practice. At present, a variety of analyzers are available that enable automated cell counting for body fluids. Nevertheless, these analyzers have some limitations and can only be operated by highly-qualified laboratory professionals. In this review, we provide an overview of the most relevant automated cell counters currently available for body fluids, the interpretation of the parameters measured by these analyzers, their main analytical features, and the role of optical microscopy as automated cell counters gain ground.
PubMed: 37363326
DOI: 10.1515/almed-2021-0011 -
Frontiers in Bioengineering and... 2019Cell identification and enumeration are essential procedures within clinical and research laboratories. For over 150 years, quantitative investigation of body fluids... (Review)
Review
Cell identification and enumeration are essential procedures within clinical and research laboratories. For over 150 years, quantitative investigation of body fluids such as counts of various blood cells has been an important tool for diagnostic analysis. With the current evolution of point-of-care diagnostics and precision medicine, cheap and precise cell counting technologies are in demand. This article reviews the timeline and recent notable advancements in cell counting that have occurred as a result of improvements in sensing including optical and electrical technology, enhancements in image processing capabilities, and contributions of micro and nanotechnologies. Cell enumeration methods have evolved from the use of manual counting using a hemocytometer to automated cell counters capable of providing reliable counts with high precision and throughput. These developments have been enabled by the use of precision engineering, micro and nanotechnology approaches, automation and multivariate data analysis. Commercially available automated cell counters can be broadly classified into three categories based on the principle of detection namely, electrical impedance, optical analysis and image analysis. These technologies have many common scientific uses, such as hematological analysis, urine analysis and bacterial enumeration. In addition to commercially available technologies, future technological trends using lab-on-a-chip devices have been discussed in detail. Lab-on-a-chip platforms utilize the existing three detection technologies with innovative design changes utilizing advanced nano/microfabrication to produce customized devices suited to specific applications.
PubMed: 31275933
DOI: 10.3389/fbioe.2019.00147 -
Turkiye Parazitolojii Dergisi May 2022Leishmaniasis is the second deadliest parasitic disease in the World Health Organisation's list of neglected diseases, following malaria. Cutaneous leishmaniasis (CL) is...
OBJECTIVE
Leishmaniasis is the second deadliest parasitic disease in the World Health Organisation's list of neglected diseases, following malaria. Cutaneous leishmaniasis (CL) is the most common form of the disease and it is one of the few communicable diseases with increasing incidence rates owing to factors like armed conflicts and climate change. CL can be divided into two major groups: Acute CL (ACL) and chronic CL (CCL). The aim of this study was to compare the efficacy of miltefosine and pentavalent antimony compounds in the CCL patient samples.
METHODS
Five isolates previously isolated from 5 CCL patients were included in this study. Genotyping is performed using internal transcribed spacer 1 (ITS 1) gene region real-time PCR. drug efficacy tests were applied to determine their activity against meglumine antimoniate (MA) and miltefosine. Serial dilutions (512, 256, 128, 64, 32, 16, 8 and 4 µg/mL) prepared from MA and miltefosine were prepared in 96-well flat-bottom cell culture plates and incubated at 24 °C for 48 hours. The efficacy of the drug on spp. promastigotes after 24 and 48 hours was evaluated by hemocytometer slide and XTT cell viability test.
RESULTS
All of the samples were genotyped as . Evaluation of 24 and 48 hours showed, 128 µg/mL and 256 µg/mL and 32 µg/mL and 64 µg/mL concentrations of miltefosine and MA were enough to kill all the promastigotes respectively. The results of the hemocytometer slide and XTT were consistent.
CONCLUSION
There are no studies investigating the efficacy of miltefosine with the CCL patient group. To overcome the treatment challenges experienced in this special patient group, more studies are needed. According to our results, it is concluded that miltefosine is efficient for the treatment of CCL and further clinical studies with miltefosine will reveal valuable data.
Topics: Antiprotozoal Agents; Humans; Leishmaniasis, Cutaneous; Meglumine Antimoniate; Phosphorylcholine
PubMed: 35604185
DOI: 10.4274/tpd.galenos.2022.85856 -
Journal of Immunology Research 2023Inflammation is closely associated with the pathogenesis of various ocular diseases. Uveitis is a condition characterized by the inflammation of the uvea and ocular...
BACKGROUND
Inflammation is closely associated with the pathogenesis of various ocular diseases. Uveitis is a condition characterized by the inflammation of the uvea and ocular tissues that causes extreme pain, decreases visual acuity, and may eventually lead to blindness. The pharmacological functions of morroniside, isolated from , are multifarious. Morroniside exerts various therapeutic effects, e.g., it ameliorates inflammation. However, the specific anti-inflammatory effect of morroniside on lipopolysaccharide-induced uveitis has not been reported widely. In this study, we investigated the anti-inflammatory effect of morroniside on uveitis in mice.
METHODS
An endotoxin-induced uveitis (EIU) mouse model was constructed and treated with morroniside. The inflammatory response was observed using slit lamp microscopy, and histopathological changes were observed by hematoxylin-eosin staining. The cell count in the aqueous humor was measured using a hemocytometer. The concentrations of TNF-, IL-6, and IL-1 in the ciliary body and retina were measured using ELISA kits. The expression of iNOS and Arg-1 in the ciliary body and retina was measured by immunofluorescence costaining, and western blotting was performed to measure the protein expression of JAK2, p-JAK2, STAT3, and p-STAT3 in the ciliary body and retina.
RESULTS
Morroniside effectively ameliorated the inflammatory response in EIU mice. Furthermore, morroniside significantly reduced the concentrations of IL-1, IL-6, and TNF- in the ciliary body and retina. Morroniside treatment significantly reduced the expression of iNOS in the ciliary body and retinal tissues. It also significantly inhibited p-JAK2 and p-STAT3 expression and promoted Arg-1 expression. In addition, morroniside boosted the effect of JAK inhibitors on the above indices.
CONCLUSIONS
Collectively, these findings suggest that morroniside may protect against LPS-induced inflammation in uveitis by promoting M2 polarization through the inhibition of the JAK/STAT pathway.
Topics: Mice; Animals; Endotoxins; Tumor Necrosis Factor-alpha; Interleukin-6; Janus Kinases; Signal Transduction; STAT Transcription Factors; Uveitis; Ciliary Body; Lipopolysaccharides; Inflammation; Anti-Inflammatory Agents; Macrophages
PubMed: 37138788
DOI: 10.1155/2023/1252873 -
Medical Science Monitor : International... Jun 2015This study aimed to investigate the relationship between miR-506 and proliferation and migration of breast cancer cells.
BACKGROUND
This study aimed to investigate the relationship between miR-506 and proliferation and migration of breast cancer cells.
MATERIAL AND METHODS
MiR-506 mimics, inhibitor, and negative control (NC) were transfected into MDA-MB-231 breast cancer cells. Cell proliferation, cell counting, colony formation assay, and Transwell assay were applied to evaluate the proliferation and migration of breast cancer cells. Data are shown as mean ± standard deviation and the experiment was performed 3 times. Statistical analyses were performed with SPSS version 10.0.
RESULTS
At 1 day after transfection, cell proliferation detected by CCK-8 assay was significantly promoted in miR-506 inhibitor when compared with the miR-506 mimics group and the NC group (P<0.05). At 3 days or 5 days after transfection, cell proliferation was markedly inhibited in the miR-506 mimics group, and miR-506 inhibitor was still significantly promoted. Cell counting with a hemocytometer showed similar results to cell proliferation. Colony formation assay showed that the number of colonies in the miR-506 mimics group was significantly smaller than that in the miR-506 inhibitor group and NC group. Transwell assay revealed that the number of migrated cells in miR-506 mimics was markedly smaller than that in the miR-506 inhibitor group and NC group.
CONCLUSIONS
MiR-506 over-expression significantly inhibits the proliferation, colony formation, and migration of breast cancer cells. miR-506 over-expression may thus be able to improve the malignant phenotype of breast cancer cells.
Topics: Breast Neoplasms; Cell Count; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Humans; In Vitro Techniques; Lipids; MicroRNAs; Molecular Mimicry; Neoplasm Metastasis; Transfection; Tumor Stem Cell Assay
PubMed: 26059632
DOI: 10.12659/MSM.893522 -
Nigerian Journal of Clinical Practice Sep 2023Neutrophils continuously migrate into the oral cavity from various sources like gingival crevicular fluid and saliva both in health and in inflammation. The migration of...
BACKGROUND
Neutrophils continuously migrate into the oral cavity from various sources like gingival crevicular fluid and saliva both in health and in inflammation. The migration of the neutrophils into the various tissues and into the oral cavity occurs when the host microbial interplay tips the balance favoring the initiation of the inflammatory and immune reactions which depending on the amount of the microbial load results in the development of acute and chronic infections in the susceptible host.
AIM
The present study was designed to quantify and compare the oral salivary neutrophil levels in patients with gingivitis and chronic and aggressive periodontitis as well as in healthy controls, before and after scaling and root planing (SRP) and to compare the difference within the selected study groups.
MATERIALS AND METHODS
Forty subjects were classified into four groups, that is, healthy controls, gingivitis, and chronic and aggressive periodontitis. Oral rinse samples were collected using Hank's balanced salt solution from each patient before and after phase I periodontal therapy. Cells in the rinse samples were stained with Acridine orange, and neutrophil counts were carried out using a fluorescence microscope and a hemocytometer.
RESULTS
Baseline oral salivary neutrophil levels were maximum in the chronic periodontitis group followed by the aggressive group and then the gingivitis group. Oral salivary neutrophil levels also positively correlated to probing pocket depth, plaque index, calculus index, and gingival index in all four study groups. Maximum reduction in the oral salivary neutrophil levels after phase I periodontal therapy was seen in the gingivitis group.
CONCLUSION
From our study, we conclude that the oral salivary neutrophil levels decreased significantly after SRP. Estimation of changes in the oral salivary neutrophil levels has the potential to aid in monitoring treatment outcomes. Thus, it suggests that it could be used as a simple, noninvasive laboratory technique to monitor the periodontal status and disease progression.
Topics: Humans; Neutrophils; Periodontal Pocket; Aggressive Periodontitis; Chronic Periodontitis; Gingivitis
PubMed: 37794540
DOI: 10.4103/njcp.njcp_3_23