Modificación de los tiempos de tinción de cortes histológicos con hematoxilina- eosina para maximizar la observación de gametos femeninos de Euthynnus. En la primera parte del estudio, se realizaron y tiñeron frotis con hematoxilina eosina (HE), Azul de Toluidina (AT), Giemsa, tinción de Wright. atrófica Atrofia glandular Metaplasia intestinal Helicobácter pylorii Tinción de Inclusiones virales Tincion hematoxilina eosina Inmunohistoquímica ++ para.
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erreverncolthyp.gq y referencias en ese trabajo). .. alizarina, la Eosina Y, el ácido carmínico, y la hematoxilina. . Teñir en hematoxilina de Delafield o de Ehrlich (el tiempo de tinción depende de las. For Teachers: Raise your hand for better lesson plans. We have the world's largest presentation database, so whether you're discussing. González Juárez Yolanda Morales Cortés Laura Patricia.
Morphology of spermatozoawiththe supra vitaldye JanusGreenwasclear, especiallythe middle piece Fig. Additionally, preparations of 10 normozoospermia patients stained with H-E, TB, Shorr and Papanicolau were assessed according to Kruger's criteria. Spermatozoa were measured in terms of 4 parameters as head length, head width, middle part length and tail length.
Statistical analysis were made using one-way variance analysis ANOVA and Post hoc Tukey HSD test and which stain showed better spermatozoa morphology and which was more appropriate for Kruger's strict criteria were determined. The smallestvalues belonged to TB according to measurements of head of the spermatozoa. Due to the use of increasing amounts of IVF in vitro fertilization techniques, studies are also focused on the morphology ofspermatozoa about the role in fertilization.
Evaluation can be done with staining spermatozoa with a variety of methods from the fresh semen by electron, fast-contrast and light microscope. There are many staining methods to evaluate themorphology of spermatozoa Uven et al. They concluded that Haematoxylin and Hemaklorun are the best staining methods for evaluation of sperm heads.
Soler et al. They evaluated spermatozoa cells stained with Hemacolor, Diff-Quik and harris haematoxylin. While they found all techniques were also useful in that used in staining of spermatozoa but Diff-Quick stain showed a significant difference. TB is used in the evaluation of the nuclear chromatin of spermatozoa by identifying the absence or rupture of disulfide bonds.
TB is a effective stain for the evaluationof changes in spermatozoa chromatin. Our study is compatible withthe knowledge of the literature.
TB dye staining method is a simple method and allow simultaneous evaluation for morphology and condensation of spermatozoa. TB is an ideal dye for routine use in this area. Chromatin condensation abnormalities of spermatozoas can be observed with Aniline Blue. The technique can be usedto assess quality of spermatozoa 7. The result for the quality of staining did not differ between the dyes.
Hammadeh et al. They examined some classical semen parameters like the chromatin condensation, spermatozoa morphology, motility and progressive motility. They claimed that there was not a correlation between the chromatin condensation and the others.
It was recommended that chromatin condense must be included, in routine laboratory observations. Spermatozoa stained with Shorr. Spermatozoa stained with Janus Green.
Morel et al. In order to show their connection with the nuclear maturity, sperms stained with Aniline Blue, Spermatozoa morphology defects differences seen between individuals and they found a significant relationship between frequent of defects and degree of nuclear maturate. Our study is compatible with the knowledge of the literature. Aniline Blue dye provides the ideal image to measure the rate of acrosome head.
Condensation and morphology can be assessed together. Estimating plasma membrane and acrosome integrity are important values for normal functions of spermatozoa, for this aim, they examined smears stained with Trypan Blue and Giemsa. They concluded that these stains can be used for the prognosis of the fertility in the semen used for IVF.
Our evaluation was consistent with the literature of Giemsa and Wright stains. Condensation and head morphology of spermatozoa were well-selectable. The middle pieceand tail could be seen. An unexpected finding that tail and head completely stained pink in the some of the spermatozoa which had damaged head morphology. A further study was suggested to explain the reason. Spermatozoa morphology and condensation were clear and smooth with Orange G stain.
But staining process was too long that's why Orange G stain had no superiority. Shorr technique is preferred in many laboratories due to the insufficient relationship in the results of IVF.
Henkel et al. The percentage of stained spermatozoa with normal and abnormal flagella's was evaluated and found a negative correlation between the stained abnormal flagella's and speed ratio and motility of the spermatozoa in elderly man. Milingos et al. They claimed that Shorr dye showed a better significant morphology than aniline blue.
Shorr is a suitable dye for morphologic evaluation of spermatozoa but it is not consideredto be appropriate for the assessment of condensation. Papanicolaou's method allows assessment of morphological of healthy human spermatozoa and also allows the separation of immature germ cells. Menkveld et al. Acrosome reaction evaluated at normal, small and large size in living and dead spermatozoa with triple stain.
Spermatozoa with small acrosome were physiologically more susceptible to cell death and loss of acrosome.
In addition, acrosome size reflects functional capacity of spermatozoa, thus the potential of male fertility can be evaluated. Spermatozoa stained with Orange Acridine were very pale and could not be evaluated. New researchesare recommended to investigate whether this image has a clinical value. Michael et al. With janus green B and tymol; head of spermatozoa, nuclear membrane, middle section was stained light green but nucleus was stained dark purple.
In this method, janus green provided an excellent staining for evaluation of spermatozoa morphology. In our study, spermatozoa heads appeared as whitish-bluish light pale coloured, middle sections and tails were stained dark blue. Our study was appeared to be compatible with the literature and as the result of our study. Spermatozoa middle section which is a rich section for mitochondria's were revealed clearly and was displayed once again by janus green.
At second phase of our study, samples of 10 patients with normazoospermatozoa stained with HE, TB, Shorr and papanicolau and analysed by Kruger strict critters. Four different measurements including head length, head width, the mild section length and the tale length were done. We've had different results than literature data. Gago et al. They have found different results for spermatozoa head measurement parameters with Haematoxylein, diff-quick Giemsa-Wright and Hemaclor stain each.
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According to this results; it is concluded that spermatozoa head measurements are affected significantly by fixation procedures and staining techniques. When we compared haematoxylin and Giemsa-Wrihgt stains as morphologically we could not find a significant difference. Our study was not compatible with the literature in this sense. Root Kustritz et al. In addition unstained samples were evaluated by using phase contrast microscope.
They found higher percentile average of spermatozoa morphologies stained by Diff-Quick as a result of their study and concluded that preparation and staining techniques can change the evaluation of spermatozoa morphology. They used Papanicolau and Diff-Quick staining methods at 20 semen samples. Manual assessments gave better results than computer assessments.
Also spermatozoa morphological evaluations were better with once washed Diff-Quick stained smears than Papanicolau stained samples. In our study; morphological assessments were better with Papanicolau stain than Giemsa and Wright stain.
In this sense our study was not compatible with literature data. In our study morphometrical measurements of spermatozoa heads were smallest with TB stained samples. Shorr and Papanicolau stains have close measurements to each other but the maximal values belonged to Papanicolau stain.
In the end we can say "for morphological assessment of spermatozoa, our special stains HE, TB, Shorr and Papanicolau are the best dyes as staining quality". Although many studies in literature are related to our work, there are only limited numbers of studies done for spermatozoa morphological assessment with comparing results of different stains. When it is thought the fact that having baby with reproduction techniques assists male infertility; the studies for morphological evaluations of spermatozoa's still remain important to select high quality spermatozoa's.
Principles of Urology. Ankara, Sun Publishing, Different fixation protocols with chromatin condensation of spermatozoa with sperm chromatin anomaly evaluation protocols. Uludag Univ. Comparison between the toluidine blue stain and the Feulgen reaction for evaluation of rabbit sperm chromatin condensation and their relationship with sperm morphology.
Theriogenology, 62 , Standardization of sampling and staining methods for the morphometric evaluation of sperm heads in the Cynomolgus monkey Macaca fascicularis using computer-assisted image analysis.
Standardization of sample preparation, staining and sampling methods for automated sperm head morphometry analysis of boar spermatozoa. Predictive value of sperm chromatin condensation aniline blue staining in the assessment of male fertility. Molecular aspects of declining sperm motility in older men.
Semen analysis. Andrology Laboratory Handbook. Istanbul, Nobel Medical Publishing, Determining sample size for the morphological assessment of sperm. Two of the reported cases proved to be positive for CMV while on an active IBD outbreak in more than one occasion during the study period cases 2 and 3.
At the time of diagnosis with CMV infection all patients were under parenteral corticoids; in 5 cases steroids were associated with azathioprine, and in 3 cases also with cyclosporine A.
No significant differences were observed between both groups when considering sex, age, type or IBD extension. We observed no significant differences with respect to oral steroid intake in the month prior to admission, or in the use of immunosuppressant drugs for the treatment of the acute IBD outbreak between the two groups. A total of 97 colonic biopsies were examined with HE and IHC, 64 of them belonged to the case group and 33 to the control group.
Practically all of these biopsies involved the left colon, and were taken from areas showing greater endoscopic abnormalities during an active IBD outbreak. The mean number of biopsies taken from patients suffering from CMV infection was 6.
In the case group, similar results were observed between both histological techniques in 49 biopsies: 14 of them being positive in both determinations and 35 being doubly negative. One of these 12 biopsies was later reclassified as positive also with HE, showing atypical inclusions.
All 33 biopsies from the control group proved negative for CMV with both techniques. The sensitivity of both histological techniques obtained from the total of 97 biopsies analyzed was The level of agreement between the results obtained in both techniques when all samples were reviewed showed a kappa value of 0.
Patients found positive for CMV were given antiviral treatment. We reported clinical improvement in 2 corticoid-resistant patients In one corticoid-resistant patient proctocolectomy was performed due to treatment failure.
Once in the body, the virus may remain latent and reactivate itself in a situation of cellular immunodepression 5. The great affinity that this virus has for organic tissues with active inflammation, combined with its high seroprevalence in patients suffering from UC, led to consider it a mere colonizer In these patients, antiviral treatment avoids the necessity for further surgical treatment and reduces mortality, thus justifying the active search for this infectious agent 2, At present, the diagnosis of CMV infection in patients suffering from IBD relies on histological techniques, since alternative methods such as serology, antigenemia, viremia, viral isolation or polymerase chain reaction PCR on blood or intestinal tissue have not proven their usefulness.
These methods do not allow differentiation between a situation of latent colonization or active infection in which CMV has a pathogenic role 1,5, The finding in colonic biopsies of the characteristic CMV changes with HE has traditionally been considered a gold standard for the diagnosis of CMV infection in these patients.
Nowadays, IHC is considered the diagnostic technique of choice, as its sensitivity in patients with serious outbreaks of IBD has been reported to be higher than the one obtained with HE 1,2, Nonetheless, these values can decrease whether IHC is carried out in colonic biopsies or in surgical specimens Moreover, its superiority compared to HE has not been established in all patients suffering from active IBD 13,19 , and there is no comparative studies between both these histological techniques.
Meaning of "hematoxilina" in the Spanish dictionary
Besides, these two histological techniques are performed routinely in different slices of the same biopsy, which makes possible the existence of cytomegalia in one slice and negativity by IHQ in the following one. A similar percentage was observed in the biopsies obtained in the 6 months prior to diagnosis in the case group patients, leading to unnecessary diagnostic delays.
On the other hand, the value obtained in this study for the Kappa index demonstrates that there is no good level of agreement when establishing a diagnosis of CMV infection between both histological techniques in colonic biopsies, as this value is lower than 0. Nevertheless, a generalized search for CMV infection in active outbreaks of IBD is unjustified due to its low prevalence 17 and the doubtful benefits of antiviral treatment in these cases.
Our data support this idea, as the benefits of antiviral treatment were only seen in steroid-refractory or steroid-dependeant IBD patients in our group. Recently, non-pathogenic genotypes of CMV have been identified, which may frequently reactivate in active IBD and spontaneously disappear without specific antiviral treatment 1,20, This new perspective allows to understand the initial consideration of CMV as a mere colonizer 7,8 and highlights the multitude of roles that this infectious agent may play in IBD.
CMV infection in patients suffering from IBD is generally considered a reactivation of latent viruses rather than a true primary infection. These patients generally have a long duration of disease and have been under multiple immunosuppressant drugs, which favor the reactivation of previous viral colonization 5,12,15, A relationship between the histological detection of CMV and previous steroidal treatment 11,17 or treatment received during an active IBD outbreak 13 has been reported.
Nevertheless, data against this fact have been published in some papers as well 10,20 , and the heterogeneity of studies carried out do not allow definitive conclusions.
Synonyms and antonyms of hematoxilina in the Spanish dictionary of synonyms
In our patients we observed an association between CMV identification in colonic tissue by IHC and the use of more than one immunosuppressant. Therefore, it could be possible that the use of stronger and combined immunosuppressive drugs in IBD may reactivate latent viruses 23 , but this topic needs to be confirmed in prospective studies. The use of combined immunosuppressants is related to a higher detection rate of the virus by IHC.
CMV still plays a controversial role in patients suffering from IBD, as the significance of this infection is not the same in all clinical settings. References 1. Kandiel A, Lashner B. Cytomegalovirus colitis complicating inflammatory bowel disease. Am J Gastroenterol ; Cytome-galovirus infection in steroid-refractory ulcerative colitis: a case-control study. Am J Surg Pathol ; Cortisone in ulcerative colitis.
Final report on therapeutic trial. Br Med J ; 2: Lennard-Jones JE. Classification of inflammatory bowel disease. Scand J Gastroenterol ; 24 Supl. Goodgame RW. Gastrointestinal cytomegalovirus disease. Ann Intern Med ; Viral investigations in ulcerative colitis and regional enteritis.
Gastroenterology ; Cytomegalovirus infection of the colon: A possible role in exacerbation of inflammatory bowel disease.
Eyre-Brook A, Dundas S. Incidence and clinical significance of colonic cytomegalovirus infection in idiopathic inflammatory bowel disease requiring colectomy.In these patients, antiviral treatment avoids the necessity for further surgical treatment and reduces mortality, thus justifying the active search for this infectious agent 2, This new perspective allows to understand the initial consideration of CMV as a mere colonizer 7,8 and highlights the multitude of roles that this infectious agent may play in IBD.
Four different measurements including head length, head width, the mild section length and the tale length were done.
At present, the diagnosis of CMV infection in patients suffering from IBD relies on histological techniques, since alternative methods such as serology, antigenemia, viremia, viral isolation or polymerase chain reaction PCR on blood or intestinal tissue have not proven their usefulness.
The great affinity that this virus has for organic tissues with active inflammation, combined with its high seroprevalence in patients suffering from UC, led to consider it a mere colonizer Una vez limpias, sacarlas de la batea e irlas situando sobre papel de filtro.
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