Salivary Glands - Science topic

Thank you very much for the answers provided by all the teachers.

Shujaat Khan

The higher percentage of malignant tumors in minor salivary glands compared to major salivary glands is due to several anatomical, histological, and biological factors:

Because of the epithelial cell component and more prone to inflammation which becomes one of the causes

Dear Doctor

Go To

"Abstract

(1) Background: Thyroid tissue ablation with radioactive iodine (RAI) has been successfully used in the treatment of differentiated thyroid cancers. However, as a side effect, RAI may induce salivary gland (SG) hypofunction, which has been alternatively managed with photobiomodulation therapy (PBMT). In our study, we assessed the effects of RAI on the SGs and further analyzed whether PBMT can minimize tissue damage. (2) Methods: Balb/c mice were allocated into three groups, as follows: RI, submitted to RAI orally; RIL, similar to RI, but with PBMT for SG hypofunction; and C, control group. The animals were euthanized on days 0, 10, and 90 after RAI. (3) Results: A decrease in tri-iodothyronine (T3) and thyroxine (T4) serum levels was observed both in the RI and RIL groups. In addition, a decrease in SG weight and morphological alterations were shown in the RI group throughout the experimental period, as well as a significant increase in total protein and peroxidase concentrations, and catalase activity. On day 90, the RI group presented less collagen and fewer sodium/iodine channels, with higher rates of cell apoptosis. Pertechnetate (Na99mTcO4) uptake was also affected in the RI group in all experimental times. Interestingly, although the RIL group also presented some alterations regarding these parameters, they were not statistically different from those of the C group on day 90. (4) Conclusions: Our results provide evidence that RAI induces harmful effects on the SGs, which can be successfully managed with PBMT."

Thank you! That is helpful. It may be trapping of air bubbles or debris, like you mentioned. We have also been vacuum sealing our unstained slides to prevent oxidation of the tissues and preserve the antigens. Maybe the suction from the vacuum sealer is causing this lifting? I would expect lifting on the edges if this was the case, so just a thought. I will have to continue troubleshooting.

@ofir zavdy thank you

pls share ur email id so that i cannot connect quickly.

Thanks Vipul, can you share some references if you have any?

More details are needed.

What are the Lanes 2&3 indicating and what were the sample differences between these lanes and samples that stacked on top of the gel?

It seems that the authors pinpointed accurately the need for a low percentage resolving gel to let the protein diffuse along the gel to be resolved. What was the largest MW of the protein at your loading ladder and which MWs are being expected from your sample?

Emir

Mr. Zeb,

For qualitative analysis of peptides, I would recommend to use vibrational spectroscopy, if the number of the components of your peptide mixture is relatively small (ca. 5-15 peptides, depending on the complexity of their molecular structure.)

There should be used, however, a set of data-processing methods and techniques for treatment of the vibrational spectroscopic patterns.

Please, consider in this context work [1] and the reference section, therein.

Because of, the qualitative analysis by means of mass spectrometry of complex mixtures of peptides does not represent s so trivial task, in particular, when there is a set of peptides with homopeptide-fragment chains. Such derivatives produce a set of structurally similar in qualitative terms fragment ions. Such fragment ions can be assigned precisely only via our quantitative method as I have already written to you.

Thus, a selection of a ''unique'' peptide by mass spectrometry (including chromatography) is significant doubtful; even it can be rather speculative, if the molecular structures of the analytes are very complex from the perspective of the number and type of the amino acid residues.

[1] Spectroscopic and structural elucidation of amino acid derivatives and small peptides: experimental and theoretical tools

Tsonko Kolev, Michael Spiteller, Bojidarka Koleva

Amino Acids, 38, 45–50 (2010)

If you have decided to use this approach: Vibrational spectroscopy to qualitative study of peptides; and if you have any questions regarding the application of the different methods and techniques for data-processing of the spectroscopic patterns, please, do not hesitate to ask them, herein. I have a significant experience in vibrational spectroscopy of peptides, in addition to a developed, my-own method for 3D structural determination via linear polarized IR-spectroscopy.

Another very similar biomarker , lipase has been extensively studied

“Lipase elevation is seen in COVID-19 and is associated with worse disease outcomes.”

Thanks again for the article, on page 3 of the paper it says:

The first tissue exposed by a bite is the skin. Intradermal

inoculation of red foxes with wild-type rabies virus

(appendix p 9) of fox origin resulted in the same median

lethal dose as the one after intramuscular inoculation.60

This indicates that skin is a possible route of rabies virus

entry. However, studies of the actual exposed neuronal

terminals through which rabies virus enters peripheral

nerves in the skin have not been done.

We did studied the deaths due to scratched without blood and published a paper:

“Scratches/Abrasions without Bleeding” Cause Rabies... in IJCM

WHO need to change the classification to only type I and III to save people.

Thanks,

Omesh

Hi Kristy, Glycerol can be used for Protein Stabilization and Prevention of Protein Aggregation. You can check this paper for your reference.

Ok, Dr Saber

Dear Giroudot,

Excellent suggestions shared already. Just wanted to add although there will be ionic imbalance after you run an electrophoresis as already ionized molecules will stick to the electrode and won't participate in the mobility. So, effectively there will be lesser ions available after each subsequent run and the elecetrophoresis might slow down due to gradual deprivation of ions.Thus, reusing should be restricted to an extent. But that should not be playing pivotal role in changing the mobility of proteins in a way to not coincide with expected molecular weight marker. Make sure your marker is running properly and you are not picking up any non specific band. It happens at times that the proteins do not match exactly the expected size according tot the molecular weight marker, but the difference should be nominal. As suggested, you should use some other proteins which have worked on earlier and see if you experience similar shifts in mobility. I reuse the lower tank buffer only once, but take fresh buffer for the upper tank every time.  Also, wash off the gel tank and electrodes properly to avoid too much accumulation of salts that might interfere with the running. 

Hope it helps!

I tried to perform the PCR a few time. However, I couldn't detect GAPDH in oral epithelial cell.  

Should I consider suitable PCR condtion? When I performed PCR, I could detect positive control. 

 In this case Two Ducts emerge from anterior border of Parotid gland instead one one. These ducts pass over masseter muscle and unite near its anterior border to form a single duct which after piercing buccinator  opens into vestibule. we found this Y- shaped pattern during dissection of a cadaver on Right side. on the left side of same cadaver the antaomy of parotid duct was normal. we published this article in International journal of Basic and Applied Sciences (Sciencepubco) and named this duct as Itoo's duct after the  Sir name of  our first author.  Anotomical Knowlege of this duct is important for surgeons operating on parotid  because leakage from this duct can lead to delayed wound healing and fistula formation in this region.

Thank you very much Ammar Mohammed. I found the link you sent helpful.

Thank you Maria, I'm trying to optimize the extraction.  I had good results yesterdays with GeneJET from Thermo.  I will also try the kit you are using and compare my results.

Ruth

Thanks so much for the responses.

Osteopontin (OPN) - multifunctional cytokine and an adhesion (glyco)protein secreted by a variety of cells, has been found to be involved in many physiologic and pathologic processes, including cancer. OPN was reported to be important for tumor initiation and progression in liver, gastric, colorectal, lung cancer. There are also data showing the participation of OPN in pathogenesis of salivary gland cancer. The exact role of OPN in carcinogenesis is not fully clarified. Some recent reports indicate the role of OPN as master regulator of Epithelial-Mesenchymal Transition (Kothari et al., 2016). According to other authors high MMP-7 and OPN expressions serve as unfavorable prognostic factors for NSCLC (Sun et al., 2015)… You can find here also the reference for two recent studies in the field of salivary gland cancer and OPN. 

1. Bjørndal K, Larsen SR, Godballe C, Krogdahl A. Osteopontin expression in salivary gland carcinomas. J Oral Pathol Med. 2011 Jul;40(6):451-5. doi: 10.1111/j.1600-0714.2010.00964.x. Epub 2010 Oct 24.

2. Fok TC, Lapointe H, Tuck AB, Chambers AF, Jackson-Boeters L, Daley TD, Darling MR. Expression and localization of osteopontin, homing cell adhesion molecule/CD44, and integrin αvβ3 in mucoepidermoid carcinoma and acinic cell adenocarcinoma of salivary gland origin. Oral Surg Oral Med Oral Pathol Oral Radiol. 2014 Sep;118(3):320-9. doi: 10.1016/j.oooo.2014.05.004. Epub 2014 May 20.

3. Sun Y, Li D, Lv XH, Hua SC, Han JC, Xu F, Li XD. Roles of osteopontin and matrix metalloproteinase-7 in occurrence, progression, and prognosis of nonsmall cell lung cancer. J Res Med Sci. 2015 Dec;20(12):1138-46. doi: 10.4103/1735-1995.172980.

4. Kothari AN, Arffa ML, Chang V, Blackwell RH, Syn WK, Zhang J, Mi Z, Kuo PC. Osteopontin-A Master Regulator of Epithelial-Mesenchymal Transition. J Clin Med. 2016 Mar 23;5(4). pii: E39. doi: 10.3390/jcm5040039.

 

 

CDC instruction in tick dissection and collection of hemolymph and saliva from engorged tick, with 8 min video 

Dear Rafik Karaman,

Thank you very much for your cooperation.

Talha

That's right. Adenos means gland. That's why carcinoma of a gland is called adenocarcinoma. Probably you have missed my point. My query is regarding the reason behind the term "adenoid", which is used in "adenoid cystic carcinoma".

If there are no contraindications, atropine i.m. for a 3 - 5 days or longer combined with the pressure dressings. In some cases it helped.

Dear Dr Leonardo Silva,

When multiple stones exist in the salivary glands, its important to look for underlying causes. The standard causes mentioned are medication intake like Diuretics and Anti-cholinergic medications, Gout and smoking. I assume that these causes have been ruled out. Hypercalcemia has been suggested in literature as a cause but you have stated that Calcium and PTH levels are normal.

I have seen a couple of patients of Sjogrens syndrome who had multiple sialolithiasis. It would be worth asking if your patient had sicca symptoms and evaluate for Sjogren's by evaluating for tear formation (Scirmer's test), testing for ANA, RF and a possible minor salivary gland biopsy. If it does indeed turn out to be Sjogrens, a variety of treatment options exist.

Hope this helps

regards

Shankar

Hello Ketaki,

You will get the best salivary gland chromosomes from climbing third instar larvae (the large fat ones that have come out of the food and are now crawling up the sides of your bottle or vial prior to forming a pupa).

Giemsa is an excellent stain to use for detecting salivary gland chromosomes.

Good luck!

You are welcome

 Best Regard

you can see hd plastic surgery on youtube.

all videos done in lyon university.they are awesome,

SEM gives you external morphology and not internal one....so thats not going to be of much use...however you can do gold dusting and try to go for maximum magnification...and check what best is visible...a better method will always be TEM for internal structure but needs a good microtome with cryopreservation....having worked shortly on animal cell lines, i would suggest you to concentrate more on mitotic proteins and pathways of salivary stem cells rather than microscopy as microscopy does not reveal much, unless u use extremely expensive florescent labelled antibodies for imaging.

Hope it helps

Regards

Yash

Dear Kashi,

I do not understand your question. Can you explain in more detail what are you planning to do with SMG tissue? Do you want to evaluate secretory function in isolated glands?

I am looking forward to hearing from you.

Best,

Marcelo

Most tumours of the salivary glands will be diagnosed with simple H-E stains but there are times when IHC will be help with diagnosis e.g poorly differentiated lesion.The following article deals with this topic:

Nagao, T., Sato, E., Inoue, R., Oshiro, H., H. Takahashi, R., Nagai, T., … Matsubayashi, J. (2012). Immunohistochemical Analysis of Salivary Gland Tumors: Application for Surgical Pathology Practice. Acta Histochemica et Cytochemica, 45(5), 269–282. doi:10.1267/ahc.12019

Hope this helps.

Since your tissue is so small and precious, you may try to measure cAMP indirectly via its effects on PKA activation. One such way would be to do your treatment, fix the tissue with PFA, slice it and do IHC for phospho-PKA substrate (antibody from Cell Signaling). This is not specific for PKA, but gives you a hint.

There are also examples in the literature for IHC of cAMP and cGMP using antibodies against these  (http://www.jbc.org/content/272/50/31489.full) and the antibodies are commercially available (http://www.antibodyresource.com/search/Antibodies/2319c8e0-28e2-529b-9030-bdb39bb25769/camp/application/IHC-P).

Good luck!

Erik

 yes, definitely, Salivary flow rate differs from children to adults. children will be having more than adults and also there are 3 types which includes serous type, mucous type and mixed type. It is believed that as the age advances salivary secretion will be reduced.

Are you asking about dissection techniques?  All insect larvae share a common bodyplan. You should be able to find a resource on dissection. Start with roaches.  A key problem with caterpillars / larvae in general is soft tissue. Ideally you will preserve them and fix beforehand.

Dear Jtim T Rainey,the best method for this patology is the cilinical examination.Abrão Rapoport

Thank you very much for your suggestions and article. It is indeed a hard task I am working on. Some papers show the separation protocol but for a different tissue which doesn't seem to work out for my tissue, so I am still looking for a solution.

Please,

I recommend you that revise all papers of Mathew Hoffman. He is an excellent resercher in this area.

Thank you for your suggestion, yes I think it might be primary antibody issue.

I mean sds-page

Hi, check link below, you might find it usefull...

OR contact Jose Ribejro at : jribeiro@niaid.nih.gov

good luck

Many systemic sources may synthesize and release the neurotrophin

in blood as epithelial cells, vascular endothelial cells,muscle cells, activate macrophages and lymphocytes. Platelets store the major part of serum BDNF: difference between plasma and serum BDNF content is 1:20.

I agree with James. If you have only two sample points (pre and post stress), I am not sure that AUC would be a good way to analyse your data. In my opinion, the option 2 is better. Other interesting analysis is the option 4, because you can also do a regression analysis with other variables (are you doing psychometric evaluations?) and try to have a model that would explain the influence of each factor in the changing of hormones due to stress.

I hope I've helped!

Best regards,

Nastassja