Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

Users Online : 30316

AbstractMaterial and MethodsResultsDiscussionConclusionReferences
Readers' Comments (0) Article in PDF Audio Visual Citation Manager Article Statistics Link to PUBMED Print this Article Send to a Friend
Advertisers Access Statistics Resources

Dr Archana Dambal

"Journal of clinical and diagnostic research is a welcome change in publishing practices. It aims to reach out to the grass-root level researchers who do not lack in experience, clinical material and ideas, but lack in their knowledge in English language and statistics. The journal achieves it's aim by supporting in these exact domains.
It also gives due credit to all research designs like descriptive and qualitative studies while many journals ignore these important study designs. The rigorous review process does not allow any compromise in quality
It is indexed in many indexing agencies and the articles are available under creative commons licence free of cost
The frequency of publication supports many aspiring authors from India and other countries.
It's wide scope welcomes articles across various specialities in medicine. In an era when there is an unscientific insistence on speciality specific research by regulatory bodies in medical education, JCDR supports collaborative research across specialities. I wish the publisher all the best in his future endeavors."



Dr. Archana Dambal
Department of General Medicine,
Belgaum Institute of Medical Sciences,Belgaum, Karnataka,INDIA,
On 30 Nov 2018




Dr Bhanu K Bhakhri

"The Journal of Clinical and Diagnostic Research (JCDR) has been in operation since almost a decade. It has contributed a huge number of peer reviewed articles, across a spectrum of medical disciplines, to the medical literature.
Its wide based indexing and open access publications attracts many authors as well as readers
For authors, the manuscripts can be uploaded online through an easily navigable portal, on other hand, reviewers appreciate the systematic handling of all manuscripts. The way JCDR has emerged as an effective medium for publishing wide array of observations in Indian context, I wish the editorial team success in their endeavour"



Dr Bhanu K Bhakhri
Faculty, Pediatric Medicine
Super Speciality Paediatric Hospital and Post Graduate Teaching Institute, Noida
On Sep 2018




Dr Mohan Z Mani

"Thank you very much for having published my article in record time.I would like to compliment you and your entire staff for your promptness, courtesy, and willingness to be customer friendly, which is quite unusual.I was given your reference by a colleague in pathology,and was able to directly phone your editorial office for clarifications.I would particularly like to thank the publication managers and the Assistant Editor who were following up my article. I would also like to thank you for adjusting the money I paid initially into payment for my modified article,and refunding the balance.
I wish all success to your journal and look forward to sending you any suitable similar article in future"



Dr Mohan Z Mani,
Professor & Head,
Department of Dematolgy,
Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



Dr Kalyani R
Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
As an experienced dentist and an academician, I proudly recommend this journal to the dental fraternity as a good quality open access platform for rapid communication of their cutting-edge research progress and discovery.
I wish JCDR a great success and I hope that journal will soar higher with the passing time."



Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




Dr. Arunava Biswas

"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."



Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011

Important Notice

Original article / research
Year : 2010 | Month : December | Volume : 4 | Issue : 6 | Page : 3450 - 3458

Histochemical Study Of Salivary Mucins In Normal And Neoplastic Salivary Glands

NAAG S* , ADI R P**

*DEPARTMENT OF ORAL AND MAXILLOFACIAL PATHOLOGY, SRI SAI COLLEGE DENTAL SURGERY, HYDERABAD, INDIA, E-MAIL: drsushma79@yahoo.com, FAX: +918416253998, PHONE: +91 9885864376; **DEPARTMENT OF ORALMAXILLOFACIAL PATHOLOGY, G. PULLA REDDY COLLEGE OF DENTAL SCIENCES, KURNOOL, INDIA, E-MAIL:drravi17@yahoo.com, PHONE: +91 9448457595

Correspondence Address :
*DEPARTMENT OF ORAL AND MAXILLOFACIAL PATHOLOGY, SRI SAI COLLEGE DENTAL SURGERY, HYDERABAD, INDIA, E-MAIL: drsushma79@yahoo.com, FAX: +918416253998, PHONE: +91 9885864376; **DEPARTMENT OF ORALMAXILLOFACIAL PATHOLOGY, G. PULLA REDDY COLLEGE OF DENTAL SCIENCES, KURNOOL, INDIA, E-MAIL:drravi17@yahoo.com, PHONE: +91 9448457595

Abstract

Background:
Salivary glands include major and minor salivary glands which are important components of the oral cavity and theirsecretions directly bathe the tissues. The biochemical and histochemical analysis of saliva revealed the presence of mucins as the main component, which showed sugar moieties and amino acids. The histochemical properties of mucins show the presence of glycoproteins andproteoglycans which differ in their chemical and structural natures. Mucins are altered in their normal and pathological states. Using special stains like Periodic Acid Schiff Reagent [PAS], Alcian Blue [AB], Aldehyde Fuchsin [AF], Mucicarmine [MC] they can be categorized into acidic, neutral, sulpho and sialomucins.

Aims:
The present study was carried out to assess different staining patterns of mucins in normal salivary glands and their neoplastic counterparts, to interpret type of mucin and change in their expression in normal and neoplastic counterparts.

Methods and Materials:
The study comprised of 19 salivary gland neoplasms which include 9 pleomorphic adenoma [PA], 4 adenoid cystic carcinoma [ADCC] and 6 mucoepidermoid carcinoma[MEC] and 10 normal salivary glands[1 parotid, 7 sub mandibular, 2 sub lingual] which were analyzed with special stains.

Results:
The results showed varied heterogeneity of mucin expression in mucous acini and change in mucin expression from benign to malignancy.

Summary and Conclusion:
The study concluded in considering the probable role of mucins in tumorigenesis and also its usage as an adjunct to diagnosis.

Keywords

mucins, salivary glands, histochemistry, special stains.

How to cite this article :

SUSHMA NAAG , RAVI PRAKASH ADI. HISTOCHEMICAL STUDY OF SALIVARY MUCINS IN NORMAL AND NEOPLASTIC SALIVARY GLANDS . Journal of Clinical and Diagnostic Research [serial online] 2010 December [cited: 2019 Sep 23 ]; 4:3450-3458. Available from
http://jcdr.net/back_issues.asp?issn=0973-709x&year=2010&month=December&volume=4&issue=6&page=3450-3458&id=1007

INTRODUCTION
Secretory cells having different distribution and morphological organizations are present with respect to the oral cavity.

They are seen from the simplest diploblastic animals to the mammals. The mammals, at the macroscopic levels, possess well developed major salivary glands viz the parotid, submaxillary and the sublingual pairs of glands. The glands are derived from the extradermal or endodermal lining of the primitive oral cavity. Saliva is the combined product of all the major and minor salivary glands. It varies qualitatively and quantitatively under normal and pathological conditions. Saliva primarily serves to moisten and lubricate food and it also assists in mastication, solubilization and partial digestion. Because of its peculiar physiochemical and immune properties, saliva maintains the homeostasis of the oral cavity and is responsible for the integrity of the oral structures. From the foregoing brief account regarding salivary glands and saliva, various studies on the nature of secretion were carried out in experimental animals with little emphasis to the salivary glands and their neoplasms in human beings. The biochemical analysis of saliva which was carried out in the early era, revealed “mucins” as the main content, which comprised of glycoproteins and oligosaccharides which were attached to sugar moieties and aminoacids. Further studies pointed out the differences in the histochemical properties of the salivary mucins of different mammalian glands and in the human salivary glands.(1),(2) Mucins are complex carbohydrates which are secreted by different types of epithelial cells and the glandular tissues of the oral cavity, the alimentary tract and the respiratory tract. Mucins reflect in their composition, the change in the functional state of the mucosa, both in the healthy and diseased states.(2)Although there have been many efforts to isolate and chemically characterize mucins, some lacunae still persist in many areas. The understanding of both the nature and the significance of mucin changes in foetal development may be potentially useful in the recognition of early neoplastic changes in adults. This has been a proven fact in the tumours of the gastrointestinal tract(2). Not many studies focus on the correlation of foetal mucins with adult content with respect to the salivary glands.

The histochemical evaluation of mucins which was elaborated by major sublingual and submandibular glands, reveal that a heterogenous population of mucosubstances exist.(3) Heterogeneity is prevalent between cells within a single acinar cluster and from acinus to acinus.(3),(4) The nature of the cartilage like substance in mixed salivary gland tumours is still widely debated and different authors have suggested that both cellular and mucinous components have an epithelial or mesenchymal origin.

Histochemically, the mucins are classified into neutral mucins and acidic mucins [which include sulpho and sialo mucins]. Many reviews on their histochemical classification and identification have been put forward to explain the intricacies of mucins.(4),[,6],(7),(8),(9) The simplest, yet a lucid method to identify mucins by routine light microscopy was employed in the present study.(4),(6),(9)

This study was undertaken in an endeavour to analyze the nature of the chondroid elements of mixed tumours and to study the changes in mucin expression in normal and neoplastic tissues by using special stains like PAS, AB at pH 1 and 2.5, Mucicarmine, the AB-PAS combination and the AF and AF-AB combination. A small attempt was made to draw a map of the mucins which were present in diseases and to find a trait which could be an aid to their morphological diagnosis, apart from its usage as an ancillary method in diagnosis.

Material and Methods

Tissue blocks were obtained from the department’s archives. A sample size of 10 normal and 10 neoplastic salivary glands, which included 9 PA, 4 ADCC and 6 MEC were taken. Normal salivary glands were obtained from the fresh cadavers and from the radical neck dissection cases which were received in the department. Normal colon and appendix samples were taken from the cadavers as controls. Due to the rarity of tumour prevalence in the geographical area where the study was done, it was not subjected to statistics.

Histochemical analysis:
Staining was performed by using Harris Hematoxylin and Eosin [S.D. Fine chemicals Ltd., Mumbai, India] for the H and E sections. MC staining was carried out by using the Southgates Mucincarmine Method. The PAS technique was employed by using the Mac Manuis [1946] method. The combined AB- PAS technique for acid and neutral mucins by [Mowry 1956] was employed. The combined AF-AB method [Spicer and Meyer 1960] was used to identify sulphated mucins and sailomucins. All the chemicals which were required for the preparation of the stains, were obtained from S.D. Fine chemicals Ltd., Mumbai, India and Qualigens Fine chemicals Ltd., Mumbai, India.

Formalin fixed, paraffin embedded tissue blocks of the biopsied specimens which were obtained, were sectioned into pieces of 5 micron thickness by using a soft tissue microtome. Special staining techniques, exclusively for mucins, were used as prescribed by the standard methodology.(4),(5),(6),(9) The slides were reviewed for “different types of mucins”, based on the staining patterns. All the slides were analyzed under a trinocular research microscope [Olympus BX 51, Japan] and the images were captured by using a 3-chip CCD Camera [Proview, Media Cybernetics, U.S.A] The results were purely descriptive in nature.

Results

A total of 19 cases of neoplastic salivary gland tumors, which included 9 benign and 10 malignant tumours respectively i.e. 9 PA, 6 MEC and 4 ADCC along with 10 normal salivary glands (2 cases of sublingual, 1 case of parotid gland 7 cases of submandibular glands), were analyzed. The goblet cells of colon and appendix were taken as controls in the study. The sections were made at 5 μ thickness, fixed to slides and were subjected to a battery of special stains including the routine H and E staining.
The serous cells were negative for all stains, although they showed strong positivity for Alcian blue. The color matching index was not similar to the standard control colon and appendix which was taken, which showed lighter intensity. As the serous acini were MC and PAS negative, the presence of mucins was ruled out with the preliminary stains. Hence, no further analysis was done to study the composition of the serous acini. The mucous acini of the submandibular and the sublingual salivary glands showed perfect heterogeneity in various areas of same acini, of different acini and between the lobules of the acini (Table/Fig 1). This was in concordance with the standard controls which were taken, thus indicating the presence of a mixture of sialomucins and sulphomucins in different areas of the mucous acini. The serous cells however could contain acid mucopolysaccharides and other highly sulfated substances (Table/Fig 1).

The PA sections showed a strong positivity for PAS and MC, which was revealed by strong magenta and red coloured staining. The myoepithelial cells were negative to all stains. The chondroid areas were highly acidophilic, indicating rich acid mucopolysaccharides. Strong AF positivity was seen in the lumen and the chondromyxoid areas, thus revealing the presence of sulphomucins.

(Table/Fig 1): Normal salivary gland with colon as control

The AB-PAS staining showed strong AB positivity in the chondromyxoid areas, thus indicating the presence of acid mucins and occasional PAS positivity in the lumen indicated the presence of neutral mucins. AF-AB staining showed an exceptionally strong reaction to AB, thus indicating a dominant presence of sialomucins. This gave the impression that the presence of sialomucins and neutral mucins was most commonly there in the benign tumour, PA (Table/Fig 2) to (Table/Fig 7).

(Table/Fig 2): Pleomorphic adenoma

(Table/Fig 3): Photomicrograph showing ductal lumen and chondromyxoid areas of PA in H&E (10X)

(Table/Fig 4): Photomicrograph showing ductal lumen and chondromyxoid areas of PA in MC (10X)


(Table/Fig 5): Photomicrograph showing ductal lumen and chondromyxoid areas of PA in AF (10X)


(Table/Fig 6): Photomicrograph showing ductal lumen and chondromyxoid areas of PA in AB-PAS (10X)

(Table/Fig 7): Photomicrograph showing ductal lumen and chondromyxoid areas of PA in AF-AB (10X)


The pseudo cystic spaces and hyaline areas of ADCC showed uniform positivity with PAS and MC, thus indicating the presence of neutral / acidic mucins. Intense acidophilia was seen with AB at different pH levels. Sulphated mucins were dominant in both the areas with AF usage. AB– PAS staining yielded no staining with PAS and moderate staining with AB, thus indicating the presence of more acidic mucins than those of the neutral type. AF- AB staining showed the presence of sulpho and sialomucins [Table /Fig 8] to [Table/Fig 13]

(Table/Fig 8): Adenoid Cystic Carcinoma


(Table/Fig 9): Photomicrograph showing pseudocystic areas and hyaline areas of ADCC in H&E along with submandibular salivary gland (low power).

(Table/Fig 10): Photomicrograph showing pseudocystic areas and hyaline areas of ADCC in MC (10X)


[Table/Fig 11]: Photomicrograph showing pseudocystic areas and hyaline areas of ADCC in AF (10X)


[Table/Fig 12]: Photomicrograph showing pseudocystic areas and hyaline areas of ADCC in AB-PAS (10X)


[Table/Fig 13] Photomicrograph showing pseudocystic areas and hyaline areas of ADCC in AF-AB (10X)

The epidermoid cells of MEC were negative to all stains. The mucous cells and lumen showed positivity to MC, PAS and AB at different pH levels, thus indicating the presence of sulphomucins. Both these areas showed a predominance of acidic mucins with AB-PAS staining and mixture of sulphomucins and sialomucins when AF-AB staining was employed. However, the stroma showed sialomucin dominance when AF- AB staining was employed and sulphated mucins when AF staining alone was done [Table /Fig 14] to [Table/ Fig 19].

[Table/Fig 14]: Mucoepdermoid Carcinoma


[Table/Fig 15]: Photomicrograph showing Mucous cells, Lumen, Epidermoid cells & stroma of MEC in H & E(10X).


[Table/Fig 16]: Photomicrograph showing Mucous cells, Lumen, Epidermoid cells & stroma of MEC in MC (10X)


[Table/Fig 17]: Photomicrograph showing Mucous cells, Lumen, Epidermoid cells & stroma of MEC in AF (10X)


[ Table/Fig 18]: Photomicrograph showing Mucous cells, Lumen, Epidermoid cells & stroma of MEC in AB-PAS (10X)


[Table/Fig 19]: Photomicrograph showing Mucous cells, Lumen, Epidermoid cells & stroma of MEC in AF-AB (10X)

The overall impression suggests the presence of sulphomucins and sialomucins in the mucous acini of the sublingual and the submandibular glands. The serous acini contain other sulfated substances in the salivary glands. The benign tumour, pleomorphic adenoma, showed a predominance of sialomucins, whereas the malignant tumours contained both sulphomucins and sialomucins.

Discussion

The mucosubstances are complex molecules, in the sense that they contain both proteins and polysaccharide components which are linked together by strong covalent chemical bonds. The mucosubstances are conveniently divided into glycoproteins and proteoglycans on the basis of their structural characteristics. (3),(10)

The glycoproteins contain one or many carbohydrate side chains of relatively small size and gross analysis shows the polypeptide chain to be the major component of the molecule. The protein-polysaccharide complexes are very different from glycoproteins and they are called as proteoglycans. They are now called as GAG and always seem to occur in the form of PG in which several polysaccharide chains are covalently linked to a polypeptide backbone.(3),(10)

The histochemical differentiation of the polysaccharide protein complex in normal and pathological tissues is of ever-increasing importance in the investigation of normal and diseased processes.(2),(11),(12),(13),(14)

Mucins are composed of a number of chemical substances which differ chemically, depending on the cell from which they are derived e.g., Epithelial mucins which include neutral mucins and acidic mucins contain sulpho and sialomucins. The connective tissue mucins show acid mucosubstances like chondroitin sulphates, keratin sulphates, hyaluronic acids and dermatan sulphates. By using different stains along with their combinations, these mucins can be categorized accordingly.(6), (7), (8),(9).

Histochemistry of the salivary gland tumours depicts the presence of the mucosubstances, but does not comment on the refined and detailed constituents of those mucosubstances, nor does it emphasize on the prognosis or longetievity of those neoplastic diseases based on the mucin pattern which has been secreted. There have been no views on the different gradations of the tumour based on pure histochemical analysis.

Mucins are complex carbohydrates which are secreted by different types of epithelial cells and glandular tissues of the respiratory and the alimentary tracts. The nature of the mucins is altered in composition from the normal to the diseased state. (12), (13), (14)

In the present study, for normal salivary glands, the serous acini were negative for all stains except AB. The mucous cells of sublingual and submandibular glands (mixed) showed diverse heterogeneity in a single acinus and in between clusters of acini, thus indicating a mixture of both acid and neutral mucins coexisting in the same gland. The mucin content in the mucous acini was composed of sulphomucins and sialomucins.

Eversole (1972) studied the mucin histochemistry in the normal salivary glands. The mucous acini of the sublingual and the submandibular glands showed a varied heterogeneity with the AB-PAS and AF stains.(15)

The present study was in concordance with the above features, with respect to the mucous acini; but the serous acini showed strong AB positivity. The results obtained from the present study coincide with the highly sulphated groups in the serous acini (which may not be the mucins), thus rendering them strongly positive for AB. As the present study did not involve enzyme histochemistry, nothing could be commented upon the loss of alcianophilia with prior enzyme treatment. Hence, probably the strong alcianoplilia of the serous acini may be attributed to the lack of enzyme usage.(16)

In this study, the chondromyxoid areas of PA showed strong positivity with the AB, PAS and MC stains. This was in accordance with the study done by Azzopardi JG and Smith OD (1959) who conducted a survey on 100 cases of salivary gland tumours.(17) The authors did not use any combination techniques for further classification. The combined AF-AB staining indicated the presence of sialomucins in these areas in the present study.

The pseudocystic spaces and hyaline areas of ADCC were positive to PAS, AB at varying pH and AF, thus indicating the presence of sulphated mucins. The AF- AB staining showed the presence of sulpho and sialomucins.

Toida et al (1985) studied 13 cases of mucinous content of ADCC by using PAS, AB 2.5, AB-PAS and other immune enzyme analyses.(18) Their results coincided with the present study in showing strong AB at pH 2.5 positivity and faint PAS staining when the AB-PAS staining technique was employed. The histochemical analysis indicated that the glycosaminoglycans in the pseudocysts were composed mainly of chondroitin sulphate and heparan sulphate.

Barnes (2001) reviewed the staining pattern of ADCC by using PAS, MC, AB and AF staining on the pseudocystic spaces and concluded that apart from PAS, MC and AB positivity; these spaces also showed AF positivity.(19), (20) This particular aspect of AF was well appreciated in the present study, which also showed strong to moderate purple colour in the pseudocystic and hyaline areas of the tumour in all the study samples.

The mucous cells and the lumen of MEC showed positivity to MC, PAS andAF staining, thus indicating the presence of sulphated mucins. The AF- AB staining showed the presence of sulpho and sialomucins in these areas. The stroma revealed a dominance of sialomucins when the AF- AB staining was employed. The epidermoid cells were negative to all stains.

Lam (1993) assessed the mucin nature between the MEC and adenosquamous carcinoma of the oesophagus by using PAS, AB, Sialidase and HID. The results showed that the MEC of oesophagus showed high amounts of sialomucins. This particular feature may be helpful in distinguishing the MEC of oesophagus from the MEC of the salivary gland, which showed high amounts of sulpho and sialomucins.(21)

In an overall view, it can be said that the present study is in concordance with the various studies which were done on human normal salivary glands and their tumours.(17),(18),(19),(20) The varied heterogeneity of the mucous acini indicated a mixture of both sulpho and sialomucins.(15) However, strong alcinophilia of the serous acini was noted in the present study, which was not seen in the previous studies.(15),(16) The possibility of the alcinophilia is attributable to a high sulphate content, although enzyme usage can cause negative alcinophilia to be seen in serous cells. Enzyme usage renders negative alcinophilia to the serous cells. The benign tumours showed sialomucins predominantly and the malignant tumours showed sulpho and silaomucins. Comparative amounts of mucins in the foetus and in inflammatory, premalignant and malignant conditions were also studied in the various areas of the body. The grading of the tumours based on the mucin production was also noted in the gastrointestinal tract. (11),(12),(13),(14) In general, a two line inference which was drawn from the study on the gastrointestinal tumours, is that they generally show sulphomucin predominance and as the differentiation of the tumour increases, the sulphated mucin content decreases with significant increase in the neutral mucin content. (2),(11),(13),(14) Such interesting and valuable findings about the salivary gland and its tumours are not sufficiently being reviewed. An extensive research in this field with more number of tissue samples will throw more light on the mucin chemistry. May be the evidence that malignant transformation develops through the sequence of changes with gradual loss of cellular differentiation and the reappearance of the foetal phenotype, can be found.

In the present study, the benign tumours showed sialomucins and as the tumours attained the malignant stage, there was an addition of sulfomucins, rendering a mixture of sulfomucins and sialomucins. Whether the change in mucin expression causes the malignant transformation or whether malignancy causes the change in mucin expression, is an issue to ponder and evaluate. This question remains speculative and debatable. The malignant salivary gland tumours showed a mucin expression which was similar to that seen in the normal mucous acini, in having a mixture of both acid mucins, but the benign tumours expressed sialomucins predominantly.

The future of mucin research lies in establishing a better understanding of the mucins in salivary glands. Such research may throw light on the structural changes of glycoproteins in precancerous lesions, which may be a promising field for histochemists and molecular biologists in knowing the exact process of carcinogenesis.

Changes in the antigenesity of glycoproteins in different diseases will be a futuristic science for immunohistochemistry and histochemistry. The genetic determinants in the expression of glycotransferase during the biosynthesis of mucins by malignant cells will be an important topic for research in the future.

Conclusion

Studies on the mucin histochemistry of salivary gland tumours are very few and are not amply reviewed. The nature or content of these mucins are not well documented. The classification of salivary gland tumours on the nature of their mucin content has not been donetill date, unlike the classifications of gastrointestinal diseases.

The nature of mucins in the gastrointestinal tract and the colorectal regions are well documented and explained. The importance of the changes in mucin expression in various regions of the tract is a known fact. The studies in gastro intestine have opened many doors for the researchers to study tumour genesis at a molecular level.

Although the present study does not aim to classify or comment on the nature of the classification, an attempt to explore the mucins and to probably understand the mucin expression in normal to neoplastic tumours has been done. It can be concluded from the present study, that there is a change in mucin expression as the tumour progresses towards malignancy. However, due to the less prevalence of salivary gland tumours in the geographical area in which this study was done, the small sample size that was studied could be the major limitation of this study. Further studies on a larger sample size may help in explaining the role of mucins in tumour pathogenesis.

The study of mucins in foetal glands and its relationship with tumorigenesis is not well established. The detailed mucin histochemistry in salivary gland tumours may give a clue about the pathophysiology of the disease, tumour differentiation, longetivity and prognostic values. The field of mucin histochemistry still remains to be further explored and and is open for research. Many hidden surprising findings might be traced regarding these tumours from their inception to a high grade malignancy. The wedding of mucin technology with immunology and lectin histochemistry is a major priority in future research. Mucin histochemistry acts as an adjunct to the routine H and E staining, but maybe helpful in assessing the malignant potentiality of tumours. A large number of tumours with various stages of malignancy and histological patterns is needed for a better understanding of the nature of mucins.

References

1.
Priya S. Comparative study of Histochemistry of salivary glands in certain mammals. PhD Anatomy Thesis, University of Poona, Maharashtra, India 1986.
2.
Ganga GM. Study of mucin histochemistry in stomach and large intestine of human fetuses and comparison with mucins of adult normal and malignant epithelial tumors. PhD Cell Biology, Thesis, Shivaji University, Maharashtra, India, 2003.
3.
Hand AR, Pathmanathan D, Field RB. Morphological features of minor salivary gland. Archs Oral Biol 1999; 44(1): 3-10
4.
Pearse EAG. Carbohydrates and Mucosubstances. In: Histochemistry theoretical and applied.Edinburgh: Churchill Livingstone: 1985. p. 675- 748.
5.
Totty BA. Mucins. In: Bancroft JD, Gamble M. editors. Theory and practice of histological techniques. 5th ed. Edinburgh: Churchill Livingstone; 2002. p.163-200.
6.
Stanley S, Rapheal SS. Staining of Carbohydrates and Connective Tissue around substance: Fibrin and Amyloid. In: Lynch Medical Laboratory Technology,3rd ed. Philadelphia; W.B. Saunders Company; 1997.p. 963-979.
7.
Drury RAB, Wallington EA. Carbohydrates. In: Carletons Histologic Technique,4th ed. New York :Mc Graw Hill Book Company.p. 1965; 201-203.
8.
Lillie RD. Polysaccharides, Mucins. In: Histopathologic technic and practical histochemistry, 3rd ed. New York. McGraw Hill Book, Company: 1965.p. 493-514.
9.
Sheehan DC, Hrapchak BB. Carbohydrates. In: Theory and practice of histotechnology.2nd ed. Columbus: Battelle Press, 1980.p. 163 – 65.
10.
Barrett AJ. The biochemistry and function of mucosubstances. Histochemical Journal 1971; 213-221.
11.
Gad A. A histochemical study of human alimentary tract mucosubstances in health and disease. Normal and tumors. British J Cancer 1969; 23(1)52-63.
12.
Lamb D, Reid L. Histochemical and autoradiographic investigation of serous cells of human bronchial glands. J Path 1970; 100:127-38.
13.
Subbuswamy SG. Pattern of mucin secretion in human intestinal mucosa. J Anat 1971;108: 291-293.
14.
Filipe MI, Branfoot AC. Abnormal patterns of mucous secretion in apparently normal mucosa of large intestine with carcinoma. Cancer 1974; 34:282-290.
15.
Eversole LR. The mucoprotein histochemistry of human mucous acinar cell containing salivary glands: Submandibular and Sublingual glands. Archs Oral Biol 1972; 17: 43-53.
16.
Baubaum CB, Jany B, Finkbeiner WE. The serous cell. Annu.Rv.Physiol 1990; 52:97-113.
17.
Azzopardi JG, Smith OD. Salivary gland tumors and their mucins. J Path Bact 1959; 77:131-140.
18.
Toida M, Takeuchi J, Sobue M, Tsukidato K, Arao S, Fukatsu T. et al. Histochemical studies on pseudocysts in adenoidcystic carcinoma of human salivary glands. Histochemical Journal 1985;17:913-924.
19.
Peel RL. Diseases of salivary glands. Barnes L. editor. Surgical Pathology of the Head and Neck. New York: Marcel Dekker Inc; 2001.
20.
Cheuk W, Chan JKC. Salivary gland tumors. In: Fletcher CDM editor. Diagnostic histopathology of tumors. 2nd ed. London: Churchill Livingstone, 2000.
21.
Lam KY, Loke SL, Ma LT. Histochemistry of mucin secreting components in mucoepidermoid and adenosquamouscarcinoma of the oesophagus. J Clin Pathol 1993; 46:1011-1015.

JCDR is now Monthly and more widely Indexed .
  • Emerging Sources Citation Index (Web of Science, thomsonreuters)
  • Index Copernicus ICV 2017: 134.54
  • Academic Search Complete Database
  • Directory of Open Access Journals (DOAJ)
  • EBSCOhost
  • Google Scholar
  • HINARI Access to Research in Health Programme
  • Indian Science Abstracts (ISA)
  • Journal seek Database
  • Google
  • Popline (reproductive health literature)
  • www.omnimedicalsearch.com