We thank Drs Rotimi and Reall for their comments, but would suggest
that their criticisms have been framed by a perspective that is different
from our own. Their perspective is the “most striking result” was that
sessile serrated adenomas (SSAs) were predominantly right sided. They are
welcome to highlight this point, which is readily apparent from the data
presented.
We thank Drs Rotimi and Reall for their comments, but would suggest
that their criticisms have been framed by a perspective that is different
from our own. Their perspective is the “most striking result” was that
sessile serrated adenomas (SSAs) were predominantly right sided. They are
welcome to highlight this point, which is readily apparent from the data
presented.
Our perspective is that the right sided predominance of SSAs is well
known, and widely reported in the literature. Hence we did not highlight
this finding. Their perspective is that a frequency of 4% of all polyps is
not “common”. From our perspective, four in every 100 polyps is “common”,
and we believe this point is worth emphasising because in many practices
SSAs are reported rarely, if ever. Again, the quantitative information is
available to readers, who are welcome to interpret it as they see fit.
We would argue that although “common” is a relative and qualitative
term, it is misleading to propose that SSAs are not common in routine
practice. Finally, Drs Rotimi and Reall are concerned that our statistical
tests may have been inappropriate. We have the happy perspective of
knowing the age distribution of the groups in question (serrated and non-
serrated polyps). We can assure them that these distributions are normal,
and that the use of parametric tests is indeed appropriate in this case.
Nicholas J Hawkins, Norman J Carr, King L Tan, Hema Mahajan, Robyn L Ward,
We fully support the integrated approach to hematopathological
diagnostics, which has been advanced by the WHO 2008 classification and we
acknowledge the prominent role of immunophenotyping in this classification
(1). The integrated approach is still not fully applied even in the most
advanced countries. In some other countries, (hemato)pathologists still
struggle, having only bone marrow (BM) tissue...
We fully support the integrated approach to hematopathological
diagnostics, which has been advanced by the WHO 2008 classification and we
acknowledge the prominent role of immunophenotyping in this classification
(1). The integrated approach is still not fully applied even in the most
advanced countries. In some other countries, (hemato)pathologists still
struggle, having only bone marrow (BM) tissue core biopsies to work with.
In some centers and countries, there may be a partial redundancy in
immunophenotyping with both flow cytometry and immunohistochemistry (IHC)
methods available.
This is not universally the case and one must rely much more on one
or the other source of information. If the histopathologists are
restricted to histology, they need to be assured that the applied IHC
methodology is accurate.
Quality assurance (QA), quality control (QC), proficiency testing, reagent
documentation and validation are standard parts of everyday practice in
clinical laboratories (2). IHC is one area of laboratory practice where
such standardization and quality assurance procedures have been addressed
relatively late (2,3). The complexity of IHC presents challenging issues
within pre-analytical, analytical, and post-analytical components, which
still make true standardization difficult. However, optimization in this
area has become possible due to rapidly advancing technologies. Also, we
have increased our understanding of each of these components, which makes
it possible to start the standardization process (3,4).
Tissue processing has been recognized as an essential pre-analytical
component that greatly determines the outcome of IHC stains (5,6). When
very few IHC markers were employed in the evaluation of BM biopsies, there
was no emphasis on QA/QC issues. Currently we face a challenge in the
light of increasing number of applied markers and the variation in tissue
processing of the BM biopsies. The results of IHC testing in BM biopsies
are not exempt from the principles that apply for any other tissue biopsy.
This means that none of the components of the IHC testing can be
neglected. Even in the situation where one type of fixative is used and no
decalcification is applied, pre-analytical components must be subjected to
the standardization process. Analytic variables such as the antigen
retrieval protocol, the detection system, and the immunostaining platform
play an important role in the quality of IHC stains.
Our pilot study included only a fraction of European Bone Marrow
Working Group laboratories. However, it disclosed that even in this
limited number of laboratories, a large variety of tissue processing and
staining methods were in use. It clearly shows that a currently used
approach to external QA/proficiency testing is not possible for BM
biopsies. At the same time our study shows that there is a need of such
external QA/QC to assure reproducibility of the results.
BM biopsy remains excluded from the good practices of QC/QA, which are
already established in surgical pathology. This is important in the light
of the increase in number of tests applied to BM biopsy, which is
happening despite advances in other, possibly more sophisticated methods
of tissue analysis. Moreover, introducing class II tests to this type of
tissue, a new landmark in BM tissue biopsy IHC is approaching.
The priority must be given to stand-alone IHC tests. However, we
disagree with Dr.Orazi that standardization and external QA/QC should be
limited to class II tests. Many of the class I tests are equally complex
to perform and may have critical impact for the final diagnosis (5). If
any clinical test deserves to be done, it also deserves to be performed
optimally and interpreted correctly. The integrated approach of WHO 2008,
at least in our minds, was never constructed to support integration of
either false-negative or false-positive IHC results or to obscure
deficiencies of technically suboptimal results.
Sincerely yours,
Emina Emilia Torlakovic, MD, PhD,
Kikkeri Naresh, M.B.B.S.; D.C.P.; M.D.;F.R.C.Path.,
Marcus Kremer, MD,
Jon van der Walt, M.D., F.R.C.Path.,
Elizabeth Hyjek, MD, PhD
Anna Porwit, MD, PhD,
References:
1. Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H,
Thiele J, Vardiman JW. WHO Classification of Tumours of Haematopoietic and
Lymphoid Tissues. WHO Press, Geneva, 2008.
2. Taylor CR. Editorial--a personal perspective. Appl Immunohistochem Mol
Morphol. 2007;15:121-3.
3. Taylor CR. Quality assurance and standardization in
immunohistochemistry. A proposal for the annual meeting of the Biological
Stain Commission, June, 1991. Biotech Histochem. 1992;67:110-7.
4. Bogen SA, Vani K, McGraw B, Federico V, Habib I, Zeheb R, Luther E,
Tristram C, Sompuram SR. Experimental validation of peptide
immunohistochemistry controls. Appl Immunohistochem Mol Morphol.
2009;17:239-46.
5. Goldstein NS, Hewitt SM, Taylor CR, Yaziji H, Hicks DG; Members of Ad-
Hoc Committee On Immunohistochemistry Standardization. Recommendations for
improved standardization of immunohistochemistry. Appl Immunohistochem Mol
Morphol. 2007;15:124-33.
6. Taylor CR, Shi S-R, Barr N, Wu N. Techniques of Immunohistochenmistry:
Principles, pitfalls, and standardization. In: Dabbs D. Diagnostic
Immunohistochemistry. Churchill Livingstone, Philadelphia, PA, USA, 2nd
Ed. 2006:17-19.
Re: Serrated and non-serrated polyps of the colorectum: their
prevalence in an unselected case series and correlation of BRAF mutation
analysis with the
diagnosis of sessile serrated adenoma
We read with interest the article by Carr NJ et al (J Clin Pathol
2009;62:516-518). The article describes the prevalence of different
colorectal polyps in an unselected population with a focus speci...
Re: Serrated and non-serrated polyps of the colorectum: their
prevalence in an unselected case series and correlation of BRAF mutation
analysis with the
diagnosis of sessile serrated adenoma
We read with interest the article by Carr NJ et al (J Clin Pathol
2009;62:516-518). The article describes the prevalence of different
colorectal polyps in an unselected population with a focus specifically on
sessile serrated adenomas (SSAs) and their correlation with BRAF mutation.
We found the most striking result in this paper presented in Table 1.
SSAs (as defined by the authors) were predominantly seen in the right side
of the colon (51 on the right and 6 on the left; a ratio of almost 9:1).
This result is all the more emphasized by the fact that not all the
patients had full colonoscopy. However, the authors failed to highlight
this important result and ultimately reported that serrated polyps were
more likely to be left-sided. This observation of a right-sided
predominance of SSAs is in agreement with many other publications(1, 2).
The finding of more right-sided SSAs have been reported to correlate with
the higher number of serrated adenocarcinomas found in the right colon in
other studies(3).
Also, the authors over-emphasize the prevalence of SSAs, concluding
that they are seen ‘commonly in routine endoscopy practice’. The SSAs
accounted for 11% of all serrated lesions and only 3.9% of all polyps in
this study and such low percentages cannot and should not be reported as
being encountered commonly in routine practice.
In addition, use of Student’s t test to analyse the relationship
between polyp type and age may have been inappropriate given the most
likely non-normal distribution of data, especially with the small numbers
in the polyp subgroups. A non-parametric statistical test would have been
more appropriate.
Olorunda Rotimi, FRCPath, PGDip Health Research,
Georgina Reall, FRCPath
References
1. Higuchi T, Sugihara K, Jass JR. Demographic and pathological
characteristics of serrated polyps of colorectum. Histopathology
2005;47:32–40.
2. Amy E. Noffsinger. Serrated Polyps and Colorectal Cancer: New
Pathway to Malignancy. Annual Review of Pathology, Mechanisms of Disease
2009;4:343–64.
3. M. J. Makinen, S. M. C. George, P. Jernvall, J. Makela, P. Vihko
and T. J. Karttunen. Colorectal carcinoma associated with serrated
adenoma - prevalence, histological features, and prognosis. Journal of
Pathology 2001; 193: 286-294
Department of Histopathology
St James's University Hospital
Leeds, UK
The authors use sweeping generalizations to make their case. This is
just one example: "while the role of external quality assurance programs
is well established as a valuable tool for QA/QC in diagnostic
immunohistochemistry in surgical pathology, such approaches are notably
lacking in bone marrow biopsy pathology". However, in reality, no
published data support a need for an external QA/QC program fo...
The authors use sweeping generalizations to make their case. This is
just one example: "while the role of external quality assurance programs
is well established as a valuable tool for QA/QC in diagnostic
immunohistochemistry in surgical pathology, such approaches are notably
lacking in bone marrow biopsy pathology". However, in reality, no
published data support a need for an external QA/QC program for bone
marrow biopsies, as proposed by the authors. A long list of unpublished
results from the NordiQC cannot be considered as equivalent to published
peer-reviewed evidence and should not have been included among the
references. Moreover, the authors themselves do not provide any data
which may suggest a need for external quality assurance for class I tests,
the only tests that they have evaluated in this study. The markers that
the authors have tested are and they should most definitely remain class I
tests. If the authors’ intent was to address the issue of analyzing
potential class II markers which can be used on bone marrow biopsy, those
are the ones they should have studied. In regard to a need for class II
markers in bone marrow biopsy evaluation, it is unclear, at least to this
reader, why immunohistology should be entitled to such a prominent place
within a complex diagnostic algorithm. Most of the neoplastic conditions
in which immunophenotyping can be useful, greatly benefit from a balanced
integrated diagnostic approach. Such an approach is the one mandated by
WHO 2008 classification. As they stand now, the conclusions that are
provide by this paper are subjective ones and most definitely not based on
evidence.
Dr. Aslam presented a rare case with peritoneal lymphomatosis and
concluded that a histologic examination is crucial in the diagnosis, while
cytology might sometimes mislead the workup.1 However, we present here
another such case with raised CA 125 and demonstrate that cytology with a
flow cytometry examination can help in the diagnosis.
This 44-year-old female suffered from autoimmune disord...
Dr. Aslam presented a rare case with peritoneal lymphomatosis and
concluded that a histologic examination is crucial in the diagnosis, while
cytology might sometimes mislead the workup.1 However, we present here
another such case with raised CA 125 and demonstrate that cytology with a
flow cytometry examination can help in the diagnosis.
This 44-year-old female suffered from autoimmune disorder and had
been using steroids for several months. She was admitted to our ward due
to progressive abdominal fullness, back soreness and shortness of breath.
An abdominal echo revealed a pelvic mass with ascites. A computed
tomography (CT) scan showed bilateral ovarian masses and multiple
lymphadenopathies over the pelvic, abdominal and retroperitoneal areas.
Biomarkers including CEA, SCC, AFP, CA 199, CA 153 were all within normal
range except for a raised CA 125 level (125.27 U/ml, normal range 2.4~36.6
U/ml).
Ascitic fluid showing a turbid appearance was aspirated and sent for
cytologic and microbiologic studies, however no micro-organisms grew from
the culture. The cytologic study revealed massive lymphocytes with bizarre
morphology. Flow cytometry was used to determine the characteristics of
the cells, and showed positive for CD10, CD19, CD20, and negative for CD7.
Under the impression of suspected lymphoma with peritoneal
lymphomatosis, CT guided biopsy of the lymph node was done. The resulting
histology examination revealed a diffuse large B cell lymphoma which was
positive for LCA, CD20 and negative for cytokeratin and CD7 (fig 1). She
received systemic chemotherapy with R-CEOP regimen, but unfortunately died
of sepsis.
Peritoneal lymphomatosis is a rare manifestation of malignant
lymphoma. Although there are some criteria that tell the difference of
peritoneal lymphomatosis from carcinomatosis, most of the cases are
indistinguishable from the clinical presentations and image studies.2,3
Even the tumor markers, such as a raised CA 125 as in this case, cannot
rule out the diagnosis of peritoneal lymphomatosis completely.4 These
factors stress the importance of putting lymphoma into the list of
diagnoses in such patients. We also highlight the important role of
cytology examinations with flow cytometry, which might also help the
physician in the diagnosis of peritoneal lymphomatosis and avoid
unnecessary studies and treatment.
1 Faculty of Medicine, College of Medicine, Kaohsiung Medical
University; and
2 Division of Hematology-Oncology, Department of Internal
Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Correspondence to: Dr SF Lin, Department of Internal Medicine,
Kaohsiung Medical University Hospital, 100, Tzu-You 1st Road, Kaohsiung
801, Taiwan.
E-mail: shlin@cc.kmu.edu.tw
Competing interests: none
REFERENCES
1. Aslam MB. Peritoneal lymphomatosis, a morphological look alike to
peritoneal carcinomatosis: a autopsy report. J Clin Pathol 2009;62:480.
2. Kim Y, Cho O, Song S, et al. Peritoneal lymphomatosis: CT findings.
Abdom Imaging 1998;23:87-90.
3. Karaosmanoglu D, Karcaaltincaba M, Oguz B, et al. CT findings of
lymphoma with peritoneal, omental and mesenteric involvement: Peritoneal
lymphomatosis. Eur J Radiol 2008; in press
4. Horger M, Muller-Schimpfle M, Yirkin I, et al. Extensive peritoneal and
omental lymphomatosis with raised CA 125 mimicking carcinomatosis: CT and
intraoperative findings. Br J Radiol 2004;77:71-3.
FIGURE LEGEND
Diffuse infiltration of medium-sized, monotonous round cells with
hyperchromatic nuclei and scant cytoplasm (H&E stain, x400). Upper right:
Positive CD20 reaction to the lymphoma cells and no reaction to
endothelial cells.
In HL-60 cells lovastatin reduces the intracellular pH in dose- dependent manner and the fall in pH correlates with the extent of DNA degradation (1). More importantly alkalinization suppresses lovostatin-induced apoptosis. A fall in pH is an indication of reductive stress, a
stimulus for the expression of HIF, VEGF and NOS. A normal or elevated pH may also be nece...
In HL-60 cells lovastatin reduces the intracellular pH in dose- dependent manner and the fall in pH correlates with the extent of DNA degradation (1). More importantly alkalinization suppresses lovostatin-induced apoptosis. A fall in pH is an indication of reductive stress, a
stimulus for the expression of HIF, VEGF and NOS. A normal or elevated pH may also be necessary for protein synthesis and cell renewal(2,3). A normal or elevated pH together with the expression of HIF (4) might, therefore, be a sine qua non for neoplastic development, growth and spread. If so how might a normal and particularly an elevated pH be
induced and apoptosis inhibited in a tissue at risk of becoming neoplastic?
The ability of adjacent tissues to support the delivery of substrate to newly vascularized tissues is clearly important in maintaining a normal pH but what if it is insufficient to maintain the adequacy of ATP resynthesis by anaerobic glycolysis, the dominant means of ATP resynthesis
in tumours? One possibility is that the pH is maintained at normal or elevated levels by the release of ammonia by cellular autophagy, apoptosis and/or necrosis. If so the elevation in pH can be expected to be restricted to a region, possibly the growing edge, interposed between
that where there is adequate substrate delivery and that in which there is protein degradation occurring in association with autophagy, apoptosis and/or necrosis. In which case the ability of contiguous tissues to sustain substrate delivery to a growth center might be the critical
variable.
Hence the question about the predictive value of VEGF-A and i-NOS expression in adjacent tissues. Might VEGF-A and i-NOS expression in unresected tissue be a better
predictor of outcome than that within the tumour, the assumption being that it is the environment, rather than the tumour per se, that determines carcinogenesis and dictates long-term survival? This possibility would have to be examined in a prospective study conducted with surgical cooperation. Which is the cart and which the horse?
Richard G Fiddian-Green
References
1. Perez-Sala D, Collado-Escobar D, Mollinedo F. Intracellular
alkalinization suppresses lovastatin-induced apoptosis in HL -60 cells
through the inactivation of a pH-dependent endonuclease. J. Biol. Chem.,
270:6235-6242, 1995.
2. P E Ballmer, M A McNurlan, H N Hulter, S E Anderson, P J Garlick,
and R Krapf. Chronic metabolic acidosis decreases albumin synthesis and
induces negative nitrogen balance in humans. J Clin Invest. 1995 January;
95(1): 39?45.
3. E Movilli, R Zani, O Carli, L Sangalli, A Pola, C Camerini, G
Cancarini, F Scolari, P Feller and R Maiorca. Correction of metabolic
acidosis increases serum albumin concentrations and decreases kinetically
evaluated protein intake in haemodialysis patients: a prospective study.
Nephrology Dialysis Transplantation, Vol 13, Issue 7 1719-1722, 1998.
4. P. H. Maxwell, G. U. Dachs, J. M. Gleadle, L. G. Nicholls, A. L.
Harris, I. J. Stratford, O. Hankinson, C. W. Pugh, and P. J. Ratcliffe.
Hypoxia-inducible factor-1 modulates gene expression in solid tumors and
influences both angiogenesis and tumor growth. PNAS July 22, 1997 vol. 94
no. 15 8104-8109.
Warthin tumor (WT) of the salivary gland (so called papillary
cystadenoma lymphomatosum or adenolymphoma) is a benign neoplasm of the
salivary gland epithelium with a proliferative epithelial component
associated with a variably prominent stroma.[1] As reported by Saxena [1],
histogenesis of this entity is very controversial. WT and malignant
lymphoma are rarely associated, and most are examples of invo...
Warthin tumor (WT) of the salivary gland (so called papillary
cystadenoma lymphomatosum or adenolymphoma) is a benign neoplasm of the
salivary gland epithelium with a proliferative epithelial component
associated with a variably prominent stroma.[1] As reported by Saxena [1],
histogenesis of this entity is very controversial. WT and malignant
lymphoma are rarely associated, and most are examples of involvement of
the lymphoid stroma of WT by a disseminated lymphoma.[1] Less commonly,
malignant lymphomas are first detected in the lymphoid stroma of WT.[1]
Most of reported cases are low-grade follicular lymphoma.[2] To the best
of our knowledge no well-documented cases of nodal marginal B-cell
lymphoma arising from WT in the same site have been described so far.
We report the case of a 74-year-old man with a fast enlarging right
parotid mass. On physical examination the mass was firm, painless,
developed in the lower pole of the parotid gland. A partial parotidectomy
was performed. At gross examination, the tumour measured 4.2 x 3.1 x 1.7
cm and cut surface revealed an irregular, brown mass replacing parotid
gland. Histologically, the tumour was composed of cystic spaces with
typical bilayered oxyphilic epithelium.(Figure 1A) This epithelium was
surrounded by malignant lymphoma characterized by diffuse infiltrate of
small to medium lymphoid cells composed of centrocyte-like B-cells,
plasmocytoid B-cells, scattered centroblast and immunoblast-like
cells.(Figure 1B) No germinal center, lympho-epithelial lesion or necrotic
areas were observed. The immunohistological study demonstrated a
phenotypical profile (CD20/CD79a/BCl-2+; CD3/CD5/CD10/CD21/cycline D1-)
consistent with marginal zone B-cell lymphoma. (Figure 1C, D)
On the basis of morphological and immunohistological findings, a
diagnosis of nodal marginal zone B-cell arising of WT's lymphoid stroma
was established.
A thorough staging examination, including cervical, thoracic and abdominal
scans revealed no evidence of other peripheral lymph nodes involvement.
All laboratory test results were normal except a small monoclonal serum
protein and mild lymphocytosis which slowly increased since 2004. The
immunophenotypage showed a monotypic B-cell lymphoid population with
moderate Lambda light chain expression. No bone marrow biopsy was done.
Because of localised disease, no treatment was started. At this time, the
patient has been followed-up for 21 months, without evidence of recurrent
tumoral syndrome.
Distribution of WT localization (parotid glands and periparotid lymph
node and its absence from other salivary tissue lacking incorporated lymph
node) seems to show that WT's histogenesis results from heteropic salivary
ductal inclusions in intra- or peri-parotid lymph nodes.[3] This theory
explains that most lymphomas arising of WT are follicular lymphomas, which
are the most frequent of systemic lymphomas.[4] No marginal zone B-cell
lymphoma arising of WT was yet reported.
Only one case reported in the same parotid gland the association of two
differents lesions: a WT and an extranodal mucosa-associated lymphoid
tissue lymphoma.[4] Extranodal marginal zone B-cell lymphoma is probably
the most common type of lymphoma truly of salivary gland origin [3] and
should be distinguished from nodal lymphomas presenting in intraparotid
lymph nodes which usually carry a worse prognosis. [5] Most of cases occur
in women, particulary aged over 50, with a history of Sjögren's syndrome
or other autoimmune disease.
In our case, no history of Sjögren's syndrome or other autoimmune disease
was mentioned. Futhermore, the absence of lympho-epithelial lesion is a
supplementary argument for the nodal origin of the marginal zone B-cell
lymphoma. This finding confirm also WT's histogenesis. Nodal marginal zone
B-cell lymphomas arising of intraparotidal Warthin's tumour are real nodal
lymphomas and require a thorough staging examination.
References
1. Saxena A., Memauri B, and Hasegawa W. Initial diagnosis of small
lymphocytic lymphoma in parotidectomy for Warthin tumour, a rare collision
tumour. J Clin Pathol. 2005;58:331-333.
2. Park CK, Manning JT, Battifora H, Medeiros LJ. Follicle center lymphoma
and Warthin tumor involving the same anatomic site. Report of two cases
and review of the literature. Am J Clin Pathol. 2000;113:113-119.
3. Marioni G, Marchese-Ragona R, Marino F et al. MALT-type lymphoma and
Warthin's tumour presenting in the same parotid gland. Acta Otolaryngol.
2004;124:318-323.
4. Isaacson PG, Müller-Hermelink HK, Piris MA et al. Extranodal marginal
zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma).
Jaffe ES, Harris NL, Stein H, Vardiman JW eds. WHO Classification Tumours
of haematopoietic and lymphoid tissues. Lyon: IARC Press, 2001:157-160.
5. Simpson RHW, Sarsfield PTL. Benign and malignant lymphoid lesions of
the salivary glands. Current Diagnostic Pathology. 1997;4:91-99.
I was very interested to read your article which was based on work we carried out a number of years ago. We feel it is important that the safety aspects of handling fish under the conditions described in the work are widely publicised.
If readers would like a more detailed account of the work carried out they will find it in:
James, S.J., James, C , Jones, S & Swain, M.J. 2000 Toxin productio...
I was very interested to read your article which was based on work we carried out a number of years ago. We feel it is important that the safety aspects of handling fish under the conditions described in the work are widely publicised.
If readers would like a more detailed account of the work carried out they will find it in:
James, S.J., James, C , Jones, S & Swain, M.J. 2000 Toxin production in fishing vessels Institute of Chemical Engineering Food & Drink 2000 Processing solutions for innovative products, Institution of Chemical Engineers, UK, ISBN 0 85295 438 7 121-124.
We would also be very interested in becoming involved in any further work that would ensure that further occurrences of the problem and the subsequent loss of life and suffering are eliminated.
We thank the reader for the careful and critical evaluation of our publication “Value of multicolor Fluorescence in situ Hybridization (UroVysionTM) in the differential diagnosis of flat urothelial lesions”
published by Stephan Schwarz, Michael Rechenmacher, Tomas Filbeck, Ruth Knuechel, Hagen Blaszyk, Arndt Hartmann and Gero Brockhoff. The reader realized some inconsistency in the Material and Methods section of our paper r...
We thank the reader for the careful and critical evaluation of our publication “Value of multicolor Fluorescence in situ Hybridization (UroVysionTM) in the differential diagnosis of flat urothelial lesions”
published by Stephan Schwarz, Michael Rechenmacher, Tomas Filbeck, Ruth Knuechel, Hagen Blaszyk, Arndt Hartmann and Gero Brockhoff. The reader realized some inconsistency in the Material and Methods section of our paper regarding HER2 immunohistochemistry (IH). In our lab for some
purposes a manual IH procedure is performed, for others we execute an automated one. In fact for the study published by Schwarz et al. we used a combination of both: section preparation was done manually and the subsequent staining procedure was performed on the NexES immunostainer
(Ventana Medical Systems, Tucson, Arizona, USA). However, we agree with the reader that we produced some inconsistency and incorrectnesses in the
Material & Methods. Hence we would like to provide this amended description of immunohistochemical HER2 staining and interpretation as follows:
“HER2 immunostaining was done on 2 mm sections using a HER2 antibody (CB11 CB11-Pathway, prediluted, FDA approved, Ventana Medical Systems, Tucson, Arizona, USA). Before staining, the tissue slides were deparaffinised and
rehydrated at room temperature. For antigen retrieval the sections were immersed in 10 mmol/l citrate buffer and microwaved for 30 min using 300 Watt (no pressure cooker was used). Subsequently the sections were cooled
by rinsing water for a couple of minutes. Then an automated staining protocol on the NexES immunostainer (Ventana Medical Systems, Tucson, Arizona, USA) was performed featuring some slight modifications with respect to the recommendation given by Ventana
(http://www.ventanamed.com/msds/files/14160USb2.pdf). The protocol described here numerously proved to work reliably on the NexES immunostainer. After a 30-minute incubation at 37°C with the primary antibody, sections were incubated for another 10 minutes at 37°C with a secondary biotinylated antibody and then with avidin-peroxidase for another 10 minutes; 3',3-diaminobenzidine was used as the chromogen. All products needed for these steps are included in the DAB detection kit provided by the manufacturer (Ventana Medical Systems, Tucson, Arizona, USA). Slides were haematoxylin counterstained, dehydrated, and mounted. A multiblock control consisting of four cell lines representing different staining intensities ranging from 0 to 3+ (MDA-MB-231: Score 0, MDA-MB-175: Score 1+, MDA-MB-453: Score 2+, and SK-BR-3: Score 3+) served as controls for all specimens.
Again we would like to thank for the careful reading of our manuscript and for coming up with the letter to the editor. This procedure assures appropriate publication quality of JCP.
Dear Editor
In the section Methods the authors described Immunohistochemistry of HER2. The described protocol is an inconsistent mixture of a manual and automated immunostaining procedure and it is very unlikely that it was actually applied.
In a second sentence the authors stated that “a manual avidin-biotin peroxidase complex procedure was used in the immunohistochemical analysis according to the ma...
Dear Editor
In the section Methods the authors described Immunohistochemistry of HER2. The described protocol is an inconsistent mixture of a manual and automated immunostaining procedure and it is very unlikely that it was actually applied.
In a second sentence the authors stated that “a manual avidin-biotin peroxidase complex procedure was used in the immunohistochemical analysis according to the manufacturer’s recommendations”. The HER 2 antibody,
CB11 from Ventana Medical Systems used in this study is recommended for staining on Ventana automated immunostainers with Ventana detection kit
and not for manual staining with other detection systems
(http://www.ventanamed.com/msds/files/14160USb2.pdf). However, if they really used the manual staining, the information about the applied detection system and the validation study reference is missing.
In contradiction with the statement in second sentence, in the fifth sentence the authors stated that “an automated staining on the Nexus (Ventana Medical Systems, Illkirch, France) was performed”. Moreover Ventana Medical Systems automated immunostainer is Nexes and not Nexus.
In the fourth sentence they stated that”the sections were microwaved in a pressure cooker” which is obvious incompatible.
In the sixth sentence they continued describing the manual immunostaining protocol – after staining (on the Nexes) they stain the sections again!
It is very interesting that many readers (authors, reviewers) overlooked such inconsistently described procedure.
Dear Editor
We thank Drs Rotimi and Reall for their comments, but would suggest that their criticisms have been framed by a perspective that is different from our own. Their perspective is the “most striking result” was that sessile serrated adenomas (SSAs) were predominantly right sided. They are welcome to highlight this point, which is readily apparent from the data presented.
Our perspective is t...
Dear Editor,
We fully support the integrated approach to hematopathological diagnostics, which has been advanced by the WHO 2008 classification and we acknowledge the prominent role of immunophenotyping in this classification (1). The integrated approach is still not fully applied even in the most advanced countries. In some other countries, (hemato)pathologists still struggle, having only bone marrow (BM) tissue...
Dear Editor,
Re: Serrated and non-serrated polyps of the colorectum: their prevalence in an unselected case series and correlation of BRAF mutation analysis with the diagnosis of sessile serrated adenoma
We read with interest the article by Carr NJ et al (J Clin Pathol 2009;62:516-518). The article describes the prevalence of different colorectal polyps in an unselected population with a focus speci...
Dear Editor
The authors use sweeping generalizations to make their case. This is just one example: "while the role of external quality assurance programs is well established as a valuable tool for QA/QC in diagnostic immunohistochemistry in surgical pathology, such approaches are notably lacking in bone marrow biopsy pathology". However, in reality, no published data support a need for an external QA/QC program fo...
Dear Editor,
Dr. Aslam presented a rare case with peritoneal lymphomatosis and concluded that a histologic examination is crucial in the diagnosis, while cytology might sometimes mislead the workup.1 However, we present here another such case with raised CA 125 and demonstrate that cytology with a flow cytometry examination can help in the diagnosis.
This 44-year-old female suffered from autoimmune disord...
Dear Editor,
In HL-60 cells lovastatin reduces the intracellular pH in dose- dependent manner and the fall in pH correlates with the extent of DNA degradation (1). More importantly alkalinization suppresses lovostatin-induced apoptosis. A fall in pH is an indication of reductive stress, a stimulus for the expression of HIF, VEGF and NOS. A normal or elevated pH may also be nece...
Dear Editor
Warthin tumor (WT) of the salivary gland (so called papillary cystadenoma lymphomatosum or adenolymphoma) is a benign neoplasm of the salivary gland epithelium with a proliferative epithelial component associated with a variably prominent stroma.[1] As reported by Saxena [1], histogenesis of this entity is very controversial. WT and malignant lymphoma are rarely associated, and most are examples of invo...
Dear Editor,
I was very interested to read your article which was based on work we carried out a number of years ago. We feel it is important that the safety aspects of handling fish under the conditions described in the work are widely publicised.
If readers would like a more detailed account of the work carried out they will find it in: James, S.J., James, C , Jones, S & Swain, M.J. 2000 Toxin productio...
We thank the reader for the careful and critical evaluation of our publication “Value of multicolor Fluorescence in situ Hybridization (UroVysionTM) in the differential diagnosis of flat urothelial lesions” published by Stephan Schwarz, Michael Rechenmacher, Tomas Filbeck, Ruth Knuechel, Hagen Blaszyk, Arndt Hartmann and Gero Brockhoff. The reader realized some inconsistency in the Material and Methods section of our paper r...
Dear Editor
In the section Methods the authors described Immunohistochemistry of HER2. The described protocol is an inconsistent mixture of a manual and automated immunostaining procedure and it is very unlikely that it was actually applied.
In a second sentence the authors stated that “a manual avidin-biotin peroxidase complex procedure was used in the immunohistochemical analysis according to the ma...
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