@ Dr. Chandan Poddar: I did not want to 'teach' you anyway....

@Prof. Deepthi Nair: thank you for your friendly assessment of my comments /contribution to the matter. I have to admit that I have not studied 'microbiology' as a subject of my university studies long ago... but due to my former profession I found those critters 'next to my eyes' on a TEM (transmission electron microscopy) screen at 3,000 - 20,000 times magnification, when they occured in my human tissue sections to be observed for a diagnostic purposes... at least at that time point I had to learn by myself how to determine and to name those tiny roundish structures... (and therefore I learned a bit about safe and successful collection, as well as about localization techniques (classical, as well as histochemical and Immunohistochemical stainings, PCR, etc...).

Last but not least: I am no Professor and quite informal person (retiree) (just the son of a 98 years old "schoolmistress" and therefore exhibiting 85% of genetic predisposition to indoctrinate others (:-)) .... Best regards, W.M.

Dear Chandan, since I lost 30 mins work in formulating a serious answer for your question and lost all due to unsaved text in the reply form within a second I only will provide the following information: If you are able to search in GOOGLE (or another data bank service/platform ) then search for | FNAC AND "tuberculous Lymphadenitis" | and you will for sure find answers to your question (see specific literature mentioned on first results page already!) [on leave] Best regards, W.M.

Thank you sir

Dear Chandan sir, you're welcome....

In my short re above in my first statement I did not want to write "since I lost 30 minutes work...." but - instead - I wanted to say "since I spent now about 30 minutes of work...). Best regards and wishes, W.M.

Thank you sir

May I ask a question back the procedure of FNAC with human technique and efforts would definitively be more expensive. ZN staunung is only a staining methid which ofcourse is cheap however cannot be compared one to one with FNAC ?

thank you madam

Apologize for lengthiness... .but perhaps this is necessry...(;-))

@ Mr./Dr. Chandan Poddar: it would be nice if you could reformulate your initial request a bit more in detail... (= what you really want instead of saying 'thanks...')

@ Mr/Mrs Prof. Dr. Deepthi Nair

Dear Ms/Mrs Prof.Dr. Deepthi Nair,

the question (text) by Chandan Poddar in itself is inconclusive.

Whilst FNAC (as I tried to point out in my lost Re#[001]) is known to be the abbreviation of a terminus technicus, namely (as you for sure know already) "Fine Needle Aspiration Cytology", the second terminus "ZN(-acid fast bacilli staining)"= Ziehl-Neelsen points to a renown and old, partially specific stain /staining method for a certain spectrum of bacilli

(@Deepthi: I know that you are familiar with these termini and the methods from your profession/profile in RG).

So I do not know so far what Mr./Dr. Chandan Poddar meant with - as you rightly pointed it out - a COMPARISON of two different assets of diagnostic techniques.. better, "techniques/methods used to end up with a diagnosis. As you (@Deepthi Nair) wrote: "ZN staining is only a staining method which of course is cheap, however cannot be compared one to one with FNAC" ? I am 100% with you and seconding that fact!

FNA/FNAB(biopsy) provides an efficient alternative to excision of tissular material which would have to be processed for either cryosectioning and/or embedding into paraffin....

In fact, to be done first is

a/the FNB (Fine Needle Biopsy), which could be either

- i) more/less an undirected punction or

- ii) an ultrasonically well directed "puncture", followed by "aspiration"), making/producing then a (histological) smear, further staining and viewing by light microscopy) to see if there are microorganisms within the cellular as well as dried fluid equivalents on the object slide.

Just as an explanation (or: reveal my thoughts) I wanted to communicate to Chandan with my first answer:

"FNAC": Usually an FNA [using suited injection needles with syringe, aspiring cellular material including fluid from the puncture target... e. g. [pending caveats!] a swollen lymphnode or a suspect tissue bulge/tumefaction/tuberous subcutaneous area] has to be done.

Smear is to be produced. Smear(s) (NB: as to my knowledge, at least usually smears on 3 separate object slides are produced and subjected to different staining methods) has to be stained accordingly:

for the entity Chandan mentioned, namely 'tuberculous lymphadenitis'

the following possibilities [with the objective of ruling out also other bacterial or microorganismic infections come to my mind:

- (for the / an experienced diagnostician: H&E)

- for the Beginner: ZN-(ZIEHL-NEELSEN) -acid fast bacilli stain

- additional Giemsa-Stain(s)

- additional other conventional microbiological methods (i.e., microbacterial cultures) and diagnostic stain for localization of bacterial infection [e.g. Gram staining technique for exclusion of any bacterial infection and/or cytological Papanicolaou staining technique on smear to detect malignant cells and other pathology )

- if there is demand due to reasonable suspicion (and 'money' or specific equipment are available) one can try also to subject the FNA-Biopsy-material to PCR.

NB: always sensibility as well as sensitivity of a method has to be taken into account! Therefore: read the papers you already have read in your project study*) as well as others which are available either by search in GOOGLE (browser or find URL www.Google.com) or - naturally also - in SCHOLAR GOOGLE!, inserting suitable keywords/search phrase ) and which deal with case studies, or review technique/methods, etc.

*) e. g. just to mention a few:

- Uwimana et al, 2016, 'Fine Needle Aspirate and Cytology (FNAC) as Useful Tool in the Diagnosis of Suspected Tuberculous Lymphadenitis in Rwanda, 2009' (URL for pdf: https://www.omicsonline.org/open-access/fine-needle-aspirate-and-cytology-fnac-as-useful-tool-in-the-diagnosis-ofsuspected-tuberculous-lymphadenitis-in-rwanda-2009-2161-1068-1000200.pdf ) NB: since this article appeared in an OMICS-published journal, one should take into consideration the reported 'predatory character' of the journal, as accused by an US court because of the suspected 'academic publisher’s deceptive business practices.' [cf. https://en.wikipedia.org/wiki/OMICS_Publishing_Group, and http://retractionwatch.com/2017/11/22/us-court-issues-injunction-omics-stop-deceptive-practices/]

- Nidhi P et al, 2011: 'FNAC in tuberculous lymphadenitis: experience from a tertiary level referral centre'. or

- Mittal P et al, 2011: 'Comparative evaluation of fine needle aspiration cytology, culture, and PCR in diagnosis of tuberculous lymphadenitis'.

or

- Aljafari AS et al, 2004: 'Diagnosis of tuberculous lymphadenitis by FNAC, microbiological methods and PCR: a comparative study'.

(now you will know mostly about what information the '30 mins of work' within my lost first reply was contained....

If there is included any wrong fact or important facts missing - @ Mr/Mrs Prof. Dr. Deepthi Nair as an expert in the field - please - feel free to comment and add valuable information from your point of view frankly and honestly...

Best wishes and regards.

lymphadenitis.13 FNAC is a well-established diagnostic

technique for lymphadenopathy evaluation. It is cost

effective, safe, minimally invasive, and a rapid method of

diagnosing not only TB lymphadenitis but also other

pathologies. It also avoids the possible physical and

psychological complications of an excision biopsy.13,14

The diagnosis of TB is still based on poorly validated symptom-based algorithms, often not resulting in a definitive diagnosis. The clinical presentation of tuberculosis is usually fever, night sweat, weight loss, anorexia. But sometimes, delay in diagnosis has often been attributed to atypical clinical presentation and radiological presentation. The diagnosis of tuberculosis by cytomorphology is not new. It is a necrotising granulomatous infection, which cytologically demonstrates the microscopic equivalent of caseous necrosis, granular-appearing necrotic background, together with mature lymphocytes, epithelioid histiocytes and multinucleated Langhans type histiocytes. Several conditions, including mycosis, bacterial and viral adenitis can present the same cytology as does mycobacterium tubercular adenitis does. Laboratory tests may be essential to establish the cause of such adenopathy correctly, because treatment and prognosis may differ. Demonstration of Mycobacterium tuberculosis in fine needle aspirates becomes necessary for an early and accurate treatment. Fine needle aspiration cytology provides a rapid and definitive tissue diagnosis in the superficial lymphadenopathy. This study demonstrates that it also permits confirmation of the presence of mycobacteria with AFB stain in microscopy. The diagnosis of tuberculosis is confirmed by the demonstration of tubercular bacilli. Mycobacteria are slender rod shaped, non-motile, non- sporing, aerobic bacterium measuring 2 to 10 um in length. India has the highest TB burden as shown in the 2011 World Health Organization (WHO) statistics.3 The diagnosis of extrapulmonary tuberculosis still remains to be more of a clinical decision. Not many clinically sensitive tests are available in India to assist the treating physician. For accurate diagnosis of M. tuberculosis, isolation and culture of organism is gold standard, but as M. tuberculosis is a slow growing organism, culture on conventional Lowenstein– Jensen medium takes 6–8 weeks. Middlebrook medium isolates growth of organism comparatively more rapidly. Mean duration to yield positive culture is about 3 weeks. But for the disease such as tuberculosis, this is too long to wait for results of culture as it is necessary to start treatment at the earliest. Therefore, comparatively rapid diagnostic strategies need to be established for diagnosis of TB

Dear Chandan, thank you for your statement which summarizes the obvious problems in rapid diagnosis of extrapulmonal TB and lacking diagnostic tools/tests in India.

Honestly added: Only a piece of advice for a good reputation in the future: if you use text formulations published already, don’t forget to cite the source(s), even if you simply write in a 'Forum/Platform' like ResearchGate.!

Example: Your text sources for:

"It is a necrotising granulomatous infection, which cytologically demonstrates the microscopic equivalent of caseous necrosis, granular-appearing necrotic background, together with mature lymphocytes, epithelioid histiocytes and multinucleated Langhans type histiocytes. Several conditions, including mycosis, bacterial and ... efficacy of fine-needle aspiration technique, ziehl-neelsen stains and ...“:

cf: MN Singh et al: - ‎2017: EFFICACY OF FINE-NEEDLE ASPIRATION TECHNIQUE, ZIEHL-NEELSEN STAINS AND CULTURE (BACTEC) IN DIAGNOSIS OF TUBERCULOUS LYMPHADENITIS IN A TERTIARY CARE HOSPITAL, GAYA, INDIA. to be found @https://www.jebmh.com/latest-articles.php?at_id=96616

and:

AHMAD et al, 2017: COMPARISON OF ZIEHL-NEELSEN STAINS WITH FINE NEEDLE ASPIRATION TECHNIQUE IN DIAGNOSIS OF TUBERCULOUS LYMPHADENITIS IN A TERTIARY CARE HOSPITAL, SOUTH BIHAR (INDIA). to be found@

https://jemds.com/latest-articles.php?at_id=13982

„It is a necrotising granulomatous infection, which cytologically demonstrates the microscopic equivalent of caseous necrosis, granular-appearing necrotic background, together with mature lymphocytes, epithelioid histiocytes and multinucleated Langhans type histiocytes. Several conditions, including mycosis, bacterial and ...“

Thank you sir

Prof. Muss, marvellous insights and advise. Grateful for all your inputs.

Thank you madam

@ Dr. Chandan Poddar: I did not want to 'teach' you anyway....

@Prof. Deepthi Nair: thank you for your friendly assessment of my comments /contribution to the matter. I have to admit that I have not studied 'microbiology' as a subject of my university studies long ago... but due to my former profession I found those critters 'next to my eyes' on a TEM (transmission electron microscopy) screen at 3,000 - 20,000 times magnification, when they occured in my human tissue sections to be observed for a diagnostic purposes... at least at that time point I had to learn by myself how to determine and to name those tiny roundish structures... (and therefore I learned a bit about safe and successful collection, as well as about localization techniques (classical, as well as histochemical and Immunohistochemical stainings, PCR, etc...).

Last but not least: I am no Professor and quite informal person (retiree) (just the son of a 98 years old "schoolmistress" and therefore exhibiting 85% of genetic predisposition to indoctrinate others (:-)) .... Best regards, W.M.

Thank you sir

Dr Muss, I did go over your immensely impressive RG profile and am sure you are far more qualified than most professors put together :)

it has been a pleasure interacting with you!

Best regards

Deepthi

Dear Prof. Dr. Deepthi Nair,

I really feel honored... - but the rules are:

"no sweat in your performance, no merits in society" (:-))

(but perhaps you have found out too that as a merely "one man performer" on /at the bench besides specific needs one can have / must develop overall knowledge and more general understanding too.... This unfortunately is not enough for being a professor in my country.... and I was not able [since there were also no premises / external presuppositions in my working contract left] - due to my duties - to teach or to give lessons at the University. I was working in a Pathology Institute of the Gen. Hospital in my city and the 'PMU' which is mentioned now as my 'affiliation' is a private medical University founded at a time when there was no possibility for a natural scientist to habilitate at a medical University... (now Hospital and PMU are joined together but are separate 'units' though). Light Microscopy, Electron Microscopy, Imaging and diagnosis of human tissue specimens was (and is) no area of expertise a beginning "medical doctor" would have been interested in (as well as the representatives of the University as well as the Hospital then...)..... Again thank you for your kind words of assessment.... but also for your valuable comment(s) here in this thread.

Perhaps we find us reading again another time when commenting or replying to an interesting question.

My best wishes and blessings, kindly yours, W.M.

NB: 2018-02-02 edited for some misspelling in and omission from the (now 'marked' by BOLD font)

Prof Dr Wolfgang H. Muss

Prof. Dr. Deepthi Nair,

Respected sir/Madam

thank you for the nice assessment of my small work in just learned in the diagnosis of M. tuberculosis is a slow growing organism, culture and Fine Needle Aspiration Cytology.

Thankfully Great person nobel 🏆 in your future Prof Dr Wolfgang H. Muss

Special thanks in Prof. Dr. Deepthi Nair great professor in India next HOD after Prof. Dr Monorama Deb

pleasure interacting with you!

Best regards

Chandan Kumar Poddar

Research Scholars

Under Supervisors

Dr S K Shahi Deans (HOD MICROBIOLOGY DEPT.)

IGIMS PATNA

Dear Chandan, you're welcome....I'm too young to get the ....🏆...(;-))

For your planning and pushing your student and future professional life I wish you all the best !

Regards W.M.

Prof.(Dr) Wolfgang H. Muss

I will conclude by thanking you so much for your kindness and generosity. I so hope and best wishes that I am able to

finished my Ph.D Work as soon possible in future, It would be wonderful with very best wishes.

Yours sincerely,

Chandan Kumar Poddar

Ph.D Research Scholar

IGIMS PATNA