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Applications of monoclonal anti-human inhibin f subunit in endometrial curettings

Leonardo D. Santos A1, James L. C. Yong A1, Xiao Juan Wu A1

Aims: Using archival material, authors studied the immunoreactivity and utility of monoclonal anti-human inhibin f subunit in the identification of chorionic villi (CV) and trophoblastic subpopulations in endometrial curettings (EC) from patients who had intra-uterine, ectopic, molar and, particularly, probable intra-uterine pregnancies. Authors also compared its expression with those of gHCG, HPL and CAM 5.2.

Methods: The four groups of EC investigated included: Group 1, 15 patients with intra-uterine pregnancies (IUP); Group 2, 15 patients with tubal pregnancies (TP); Group 3, 15 patients with hydatidiform moles (HM); and Group 4, 20 patients with purported history of intra-uterine pregnancies (PIUP). Positive and negative control cases were from Groups 1 and 3 and Group 2, respectively. The test cases were from Group 4. Immunohistochemistry was performed on each case testing for expression of inhibin f, gHCG, HPL and CAM 5.2.

Results: Trophoblastic populations, which included syncytiotrophoblast (ST), cytotrophoblast (CT) and intermediate trophoblast (IT), were absent in all 15 negative control cases (Group 2). The 30 positive control cases (Groups 1 and 3) revealed the following: (a) ST, CT and IT were identified in all cases and were positive for CAM 5.2, (b) inhibin f, gHCG and HPL (except one case) were reactive for all cases with ST, but not CT, and (c) IT positivity for gHCG, HPL and inhibin f was 67, 80-93 and 100%, respectively. From the 20 test cases (Group 4), the findings were: (a) CT was absent in all cases, (b) scattered ST cells, which were identified only in 10 cases, were positive for all antibodies, (c) scattered IT cells were present in 17 cases and showed 100% CAM 5.2 positivity, and (d) IT positivity for gHCG, inhibin f and HPL was 58.8% (10/17), 76.5% (13/17) and 82.4% (14/17), respectively. Background staining was observed in 22 of 65 cases (33.8%) stained with gHCG and HPL; half of these cases came from Group 3. Inhibin f and CAM 5.2 staining did not show this problem.

Conclusions: Authors suggest that inhibin f is a useful antibody in diagnosing IUP and HM and in documenting intra-uterine gestations in cases with PIUP because it is a sensitive marker in immunolabelling IT and ST. Combined application of inhibin f and CAM 5.2 might be more useful than gHCG and HPL because the latter showed background staining in one third of the cases.

Pathology 35(3): 217-223, 2003

Apoptosis, Proliferation, and Expression of p53 and bcl-2 in Endocervical Glandular Intraepithelial Lesions and Invasive Endocervical Adenocarcinoma.

Ali-Fehmi R, Qureshi F, Lawrence WD, Jacques SM

Authors evaluated apoptosis, proliferation, and p53 and bcl-2 expression in a spectrum of intraepithelial and invasive endocervical glandular lesions currently recognized by the World Health Organization as adenocarcinoma in situ, lesions with atypia "less than adenocarcinoma in situ" (endocervical glandular dysplasia and endocervical glandular atypia), and invasive adenocarcinoma. Aside from nuclear atypia, increased mitotic activity and apoptosis are consistent and closely correlated morphologic features of endocervical adenocarcinoma in situ. Apoptotic bodies and mitotic figures were counted in 32 examples of normal endocervical glands, 35 of endocervical glandular atypia, 30 of endocervical glandular dysplasia, 34 of adenocarcinoma in situ, and 30 of invasive adenocarcinoma. These results were correlated with immunohistochemical staining for MIB1, bcl-2, and p53 performed on 20 examples of each. Mitotic counts, p53 expression, and bcl-2 expression all increased significantly and in proportion to the degree of atypia in the spectrum of endocervical lesions. Apoptotic body counts and MIB1 expression also increased significantly with increasing atypia, but showed higher levels in adenocarcinoma in situ than in invasive adenocarcinoma. Apoptosis correlates with proliferation as measured by mitotic counts and MIB1, and also with p53 and bcl-2 expression. Apoptosis appears to be an important mechanism in the pathogenesis of endocervical glandular lesions and may be useful as an aid in their evaluation and diagnosis.

Int J Gynecol Pathol. 2004 Jan; 23(1): 1-6.

SARS case in lab worker

The World Health Organization has confirmed that it has received a report from Taipei that a 44-year-old male lab worker, working in a military laboratory has been infected with severe acute respiratory syndrome (SARS). The news has raised fears once again that another outbreak of the disease could originate from a lab. Tests conducted in two Taiwan laboratories confirmed the presence of SARS coronavirus in the lab worker. The lab is the only level P4 lab in Taiwan. At the moment, this appears to be an isolated incident, but all contacts are being followed. This incident illustrates that SARS presents a continuing threat. In the post-epidemic period, the greatest risk from SARS may be through exposure in laboratories where the virus is used or stored, WHO said in a statement. For this reason, WHO urges countries to conduct an inventory of laboratories and samples that they hold and to ensure that the correct biosafety procedures are being followed, as approved at a WHO laboratory workshop on 22 October, 2003.

The Scientist, Dec. 17, 2003

Results from laboratory tests have confirmed a case of SARS in a 32-year-old man in the southern Chinese province of Guangdong. The patient is a television producer who has been under treatment, in isolation, at a hospital in the provincial capital, Guangzhou, since December 20. This is the first confirmed case of SARS in 2004, and the first case not linked to a laboratory accident that has occurred since the initial outbreak of SARS was declared contained on July 5, 2003.

Interpretation of diagnostic laboratory tests for severe acute respiratory syndrome: the Toronto experience

Background: An outbreak of severe acute respiratory syndrome (SARS) began in Canada in February 2003. The initial diagnosis of SARS was based on clinical and epidemiological criteria. During the outbreak, molecular and serologic tests for the SARS associated coronavirus (SARS-CoV) became available. However, without a gold standard, it was impossible to determine the usefulness of these tests. Authors describe how these tests were used during the first phase of the SARS outbreak in Toronto and offer some recommendations that may be useful if SARS returns.

Methods: Authors examined the results of all diagnostic laboratory tests used in 117 patients admitted to hospitals in Toronto who met the Health Canada criteria for suspect or probable SARS. Focusing on tests for SARS-CoV, we attempted to determine the optimal specimen types and timing of specimen collection.

Results: Diagnostic test results for SARS-CoV were available for 110 of the 117 patients. SARS-CoV was detected by means of reverse-transcriptase polymerase chain reaction (RT-PCR) in at least one specimen in 59 (54.1%) of 109 patients. Serologic test results of convalescent samples were positive in 50 (96.2%) of 52 patients for whom paired serum samples were collected during the acute and convalescent phases of the illness. Of the 110 patients, 78 (70.9%) had specimens that tested positive by means of RT-PCR, serologic testing or both methods. The proportion of RT-PCR test results that were positive was similar between patients who met the criteria for suspect SARS (50.8%, 95% confidence interval [CI] 38.4%63.2%) and those who met the criteria for probable SARS (58.0%, 95% CI 44.2% 70.7%). SARS-CoV was detected in nasopharyngeal swabs in 33 (32.4%) of 102 patients, in stool specimens in 19 (63.3%) of 30 patients, and in specimens from the lower respiratory tract in 10 (58.8%) of 17 patients.

Interpretation: These findings suggest that the rapid diagnostic tests in use at the time of the initial outbreak lack sufficient sensitivity to be used clinically to rule out SARS. As tests for SARS-CoV continue to be optimized, evaluation of the clinical presentation and elucidation of a contact history must remain the cornerstone of SARS diagnosis. In patients with SARS, specimens taken from the lower respiratory tract and stool samples test positive by means of RT-PCR more often than do samples taken from other areas.

Objective: To evaluate the role of bacterial antigen detection test in cerebrospinal fluid (CSF) for a rapid etiological diagnosis of bacterial meningitis.

Methods: The study included 36 cases of bacterial meningitis and 14 controls. Latex particle agglutination test (LPA test) for detection of bacterial antigen was done in the CSF using slidex meningitis kit (Biomeriux, France).

Results: Using LPA test, an etiological diagnosis could be made in 83% cases of bacterial meningitis. In contrast, CSF Gram stain and culture showed 36% and 6% positivity, respectively. The sensitivity and specificity of LPA test were 83% and 100%, respectively. The common etiological organisms were S. pneumoniae, H. influenzae type b and N. meningitidis A. S. pneumoniae was encountered in all age groups while H. influenzae type b was found only below one year of age.

Conclusions: LPA test is a rapid and superior diagnostic tool as compared to CSF Gram stain and culture. The study recommends LPA test as an adjunct laboratory test for rapid etiological diagnosis of bacterial meningitis for prompt institution of proper antibiotics.


		January 2004
		Applications of monoclonal anti-human inhibin f subunit in endometrial curettings Leonardo D. Santos A1, James L. C. Yong A1, Xiao Juan Wu A1 Aims: Using archival material, authors studied the immunoreactivity and utility of monoclonal anti-human inhibin f subunit in the identification of chorionic villi (CV) and trophoblastic subpopulations in endometrial curettings (EC) from patients who had intra-uterine, ectopic, molar and, particularly, probable intra-uterine pregnancies. Authors also compared its expression with those of gHCG, HPL and CAM 5.2. Methods: The four groups of EC investigated included: Group 1, 15 patients with intra-uterine pregnancies (IUP); Group 2, 15 patients with tubal pregnancies (TP); Group 3, 15 patients with hydatidiform moles (HM); and Group 4, 20 patients with purported history of intra-uterine pregnancies (PIUP). Positive and negative control cases were from Groups 1 and 3 and Group 2, respectively. The test cases were from Group 4. Immunohistochemistry was performed on each case testing for expression of inhibin f, gHCG, HPL and CAM 5.2. Results: Trophoblastic populations, which included syncytiotrophoblast (ST), cytotrophoblast (CT) and intermediate trophoblast (IT), were absent in all 15 negative control cases (Group 2). The 30 positive control cases (Groups 1 and 3) revealed the following: (a) ST, CT and IT were identified in all cases and were positive for CAM 5.2, (b) inhibin f, gHCG and HPL (except one case) were reactive for all cases with ST, but not CT, and (c) IT positivity for gHCG, HPL and inhibin f was 67, 80-93 and 100%, respectively. From the 20 test cases (Group 4), the findings were: (a) CT was absent in all cases, (b) scattered ST cells, which were identified only in 10 cases, were positive for all antibodies, (c) scattered IT cells were present in 17 cases and showed 100% CAM 5.2 positivity, and (d) IT positivity for gHCG, inhibin f and HPL was 58.8% (10/17), 76.5% (13/17) and 82.4% (14/17), respectively. Background staining was observed in 22 of 65 cases (33.8%) stained with gHCG and HPL; half of these cases came from Group 3. Inhibin f and CAM 5.2 staining did not show this problem. Conclusions: Authors suggest that inhibin f is a useful antibody in diagnosing IUP and HM and in documenting intra-uterine gestations in cases with PIUP because it is a sensitive marker in immunolabelling IT and ST. Combined application of inhibin f and CAM 5.2 might be more useful than gHCG and HPL because the latter showed background staining in one third of the cases. Pathology 35(3): 217-223, 2003 Apoptosis, Proliferation, and Expression of p53 and bcl-2 in Endocervical Glandular Intraepithelial Lesions and Invasive Endocervical Adenocarcinoma. Ali-Fehmi R, Qureshi F, Lawrence WD, Jacques SM Authors evaluated apoptosis, proliferation, and p53 and bcl-2 expression in a spectrum of intraepithelial and invasive endocervical glandular lesions currently recognized by the World Health Organization as adenocarcinoma in situ, lesions with atypia "less than adenocarcinoma in situ" (endocervical glandular dysplasia and endocervical glandular atypia), and invasive adenocarcinoma. Aside from nuclear atypia, increased mitotic activity and apoptosis are consistent and closely correlated morphologic features of endocervical adenocarcinoma in situ. Apoptotic bodies and mitotic figures were counted in 32 examples of normal endocervical glands, 35 of endocervical glandular atypia, 30 of endocervical glandular dysplasia, 34 of adenocarcinoma in situ, and 30 of invasive adenocarcinoma. These results were correlated with immunohistochemical staining for MIB1, bcl-2, and p53 performed on 20 examples of each. Mitotic counts, p53 expression, and bcl-2 expression all increased significantly and in proportion to the degree of atypia in the spectrum of endocervical lesions. Apoptotic body counts and MIB1 expression also increased significantly with increasing atypia, but showed higher levels in adenocarcinoma in situ than in invasive adenocarcinoma. Apoptosis correlates with proliferation as measured by mitotic counts and MIB1, and also with p53 and bcl-2 expression. Apoptosis appears to be an important mechanism in the pathogenesis of endocervical glandular lesions and may be useful as an aid in their evaluation and diagnosis. Int J Gynecol Pathol. 2004 Jan; 23(1): 1-6. SARS case in lab worker The World Health Organization has confirmed that it has received a report from Taipei that a 44-year-old male lab worker, working in a military laboratory has been infected with severe acute respiratory syndrome (SARS). The news has raised fears once again that another outbreak of the disease could originate from a lab. Tests conducted in two Taiwan laboratories confirmed the presence of SARS coronavirus in the lab worker. The lab is the only level P4 lab in Taiwan. At the moment, this appears to be an isolated incident, but all contacts are being followed. This incident illustrates that SARS presents a continuing threat. In the post-epidemic period, the greatest risk from SARS may be through exposure in laboratories where the virus is used or stored, WHO said in a statement. For this reason, WHO urges countries to conduct an inventory of laboratories and samples that they hold and to ensure that the correct biosafety procedures are being followed, as approved at a WHO laboratory workshop on 22 October, 2003. The Scientist, Dec. 17, 2003 Laboratory Confirmation of SARS Case in Southern China Results from laboratory tests have confirmed a case of SARS in a 32-year-old man in the southern Chinese province of Guangdong. The patient is a television producer who has been under treatment, in isolation, at a hospital in the provincial capital, Guangzhou, since December 20. This is the first confirmed case of SARS in 2004, and the first case not linked to a laboratory accident that has occurred since the initial outbreak of SARS was declared contained on July 5, 2003. World Health Organisation, Jan. 6, 2004 Interpretation of diagnostic laboratory tests for severe acute respiratory syndrome: the Toronto experience Patrick Tang, Marie Louie, Susan E. Richardson, et al. Background: An outbreak of severe acute respiratory syndrome (SARS) began in Canada in February 2003. The initial diagnosis of SARS was based on clinical and epidemiological criteria. During the outbreak, molecular and serologic tests for the SARS associated coronavirus (SARS-CoV) became available. However, without a gold standard, it was impossible to determine the usefulness of these tests. Authors describe how these tests were used during the first phase of the SARS outbreak in Toronto and offer some recommendations that may be useful if SARS returns. Methods: Authors examined the results of all diagnostic laboratory tests used in 117 patients admitted to hospitals in Toronto who met the Health Canada criteria for suspect or probable SARS. Focusing on tests for SARS-CoV, we attempted to determine the optimal specimen types and timing of specimen collection. Results: Diagnostic test results for SARS-CoV were available for 110 of the 117 patients. SARS-CoV was detected by means of reverse-transcriptase polymerase chain reaction (RT-PCR) in at least one specimen in 59 (54.1%) of 109 patients. Serologic test results of convalescent samples were positive in 50 (96.2%) of 52 patients for whom paired serum samples were collected during the acute and convalescent phases of the illness. Of the 110 patients, 78 (70.9%) had specimens that tested positive by means of RT-PCR, serologic testing or both methods. The proportion of RT-PCR test results that were positive was similar between patients who met the criteria for suspect SARS (50.8%, 95% confidence interval [CI] 38.4%63.2%) and those who met the criteria for probable SARS (58.0%, 95% CI 44.2% 70.7%). SARS-CoV was detected in nasopharyngeal swabs in 33 (32.4%) of 102 patients, in stool specimens in 19 (63.3%) of 30 patients, and in specimens from the lower respiratory tract in 10 (58.8%) of 17 patients. Interpretation: These findings suggest that the rapid diagnostic tests in use at the time of the initial outbreak lack sufficient sensitivity to be used clinically to rule out SARS. As tests for SARS-CoV continue to be optimized, evaluation of the clinical presentation and elucidation of a contact history must remain the cornerstone of SARS diagnosis. In patients with SARS, specimens taken from the lower respiratory tract and stool samples test positive by means of RT-PCR more often than do samples taken from other areas. CMAJ 2004; 170(1): 47-54 Bacterial antigen detection test in meningitis Das BK, Gurubacharya RL, Mohapatra TM, Mishra OP Department of Pediatrics, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India. Objective: To evaluate the role of bacterial antigen detection test in cerebrospinal fluid (CSF) for a rapid etiological diagnosis of bacterial meningitis. Methods: The study included 36 cases of bacterial meningitis and 14 controls. Latex particle agglutination test (LPA test) for detection of bacterial antigen was done in the CSF using slidex meningitis kit (Biomeriux, France). Results: Using LPA test, an etiological diagnosis could be made in 83% cases of bacterial meningitis. In contrast, CSF Gram stain and culture showed 36% and 6% positivity, respectively. The sensitivity and specificity of LPA test were 83% and 100%, respectively. The common etiological organisms were S. pneumoniae, H. influenzae type b and N. meningitidis A. S. pneumoniae was encountered in all age groups while H. influenzae type b was found only below one year of age. Conclusions: LPA test is a rapid and superior diagnostic tool as compared to CSF Gram stain and culture. The study recommends LPA test as an adjunct laboratory test for rapid etiological diagnosis of bacterial meningitis for prompt institution of proper antibiotics. Indian J Pediatr. 2003: 70(10): 799-801.
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