Immunology of Infectious Diseases - Science topic

Prolonged viral shedding refers to the continued presence of influenza virus in respiratory secretions beyond the typical duration observed in immunocompetent adults. In immunocompetent individuals, influenza viral shedding usually lasts for about 5 to 7 days after symptom onset but may persist for longer periods in some cases. Prolonged shedding in immunocompetent adults may be defined as shedding persisting beyond 10 days after symptom onset.

In immunosuppressed individuals, such as those with HIV/AIDS or undergoing immunosuppressive therapy, viral shedding can be prolonged further due to compromised immune responses. In these cases, prolonged shedding may be defined as viral presence beyond 14 days after symptom onset. However, the exact definition of prolonged viral shedding can vary depending on the specific context, such as the severity of illness, the presence of complications, and individual patient factors.

There is no commercially available vaccine against schistosomiasis. However, there have been ongoing efforts to develop one. Several vaccine candidates have shown promise in preclinical and early clinical trials.One of the most advanced vaccine candidates is the Sm-p80-based vaccine, which targets Schistosoma mansoni. Sm-p80 is a protein involved in the penetration of schistosome larvae through host skin. Experimental trials of this vaccine have shown significant reductions in parasite burden and protection against infection in animal models. Clinical trials in humans have also demonstrated safety and some degree of efficacy, but further research is needed to optimize its effectiveness.Other potential vaccine candidates include antigens derived from other stages of the schistosome life cycle, such as cercariae and adult worms. These vaccines aim to stimulate the host immune system to produce protective antibodies and immune responses that prevent infection or reduce parasite burden upon infection.

Transduction of T cells with adeno-associated virus (AAV) vectors involves introducing a gene of interest into T cells for various therapeutic purposes. This process includes selecting an appropriate AAV vector, cloning the gene of interest into the vector, producing recombinant AAV, transducing T cells ex vivo or in vivo, culturing and expanding transduced T cells, and applying them for specific therapeutic purposes such as cancer immunotherapy or gene therapy for genetic diseases. Monitoring and evaluation are crucial for assessing safety, efficacy, and potential side effects.

What rapid tests do you recommend to detect Mayaro virus?

Have a look at these useful links.

PRNT stands for plaque-reduction neutralization titers. We use a plaque assay to determine the level of infectious virus in a solution using 10-fold serial dilutions of the volume. We pick a culture dish with a moderate # of plaques which can be counted, then take the reciprocal to get a plaque titer.

PRNT measures how much you can dilute your antibodies and still reduce the plaque # by 50%.

Hi, did you solve the problem? could you please share some experience?

Thanks for all the tips! This was extremely helpful

Thanks for all the tips! This was extremely helpful

Thanks for all the tips! This was extremely helpful

Hi. Mostly aphthous ulcers, can be a result of stress, certain vitamin deficiency, hormonal fluctuations and even unintended mouth injuries.

It's well-known that most Gram-negative bacteria are intrinsically resistant to vancomycin because their outer membranes are impermeable to large glycopeptide molecules (with the exception of some non-gonococcal Neisseria species).

Regards

Here is a diagram of murine lymph nodes. Also an article that explains a method of locating them for the first time. Good luck!

For trends analysis purpose, it is recommended to use the age-standardize YLL rate instead of YLL rate. This is important because the age structure of the population changes over time, so it requires to adjust for age as a confounder factor.

I hope this helps.

Hi,

Can you test the acridine orange for coloration of DNA ?

 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC120052/

I understand you wanted to use TDF/XTC or TAF/XTC  in your patient given the activity against both HIV and HBV. The fact is that you can use either of those meds on a fixed-dose combination or individual drug combinations. If you cannot use TDF alone, use entecavir in addition to a fully suppressive antiretroviral regimen but do not use duplicate therapy with 3TC.

Now, my real worry about your question is if you studied the failure? The failure in this case would likely be explained by resistance to EFV but confirmation is needed. What about liver, renal or bone toxicity in your patient?

Priya we use Working Conc 1µg/ml in our lab to induce inflammation in cell culture

(Keep LPS Stock Conc 1mg/ml@4degree)

Hope this helps

we had a case which was managed conservatively by medical management, want to make a report and publish our experience. Do you have any suggestions regarding journals which would be interested in such a case report ??

Looks like TLR4 inhibitors like Metformin can resolve some of interferon's side effects.

Dear Joshua Adekunle Babalola

Pls. find the attached files

Brian K. Nunnally, Ira S. Krull. 2003. Prions and Mad Cow Disease. CRC Press,Pp.413. ISBN0203912977, 9780203912973

Hoping this will be helpful            

Regards

Prof. Houda Kawas

For how long did you give heat shock to the DH5alpha cells? It is recommended to keep your competent cells always in cold ice after taking then out from the -80 degree centigrade freezer. After adding your recombinant DNA ( ligated to your working plasmid) to the competent cells, incubate for a short while in ice. Then give heat shock by placing the vial in water bath at 42 degree centigrade for 45 seconds only. Immediately put back the vial in ice and keep for 5 minutes. Then add LB broth (without any antibiotic) and keep in 37 degree incubator shaker for 1 hour and then plate on your agar plates containing appropriate antibiotic.

Bacterial population increase i.e. growth is measured by checking OD at 600nm. With experience even by sight you can make rough working estimations.

When you get colonies, just proceed for making glycerol cell stock of those, and also isolate plasmid and send for sequencing to confirm presence of desired insert.

A biomarker actually very useful to study glioblastoma is TLR3. Little studied, but it has a unique theranostic value.

Because the target of TLR3, which is poly IC, has an amazing tumoricidal activity.

Several similar to poly IC, such as Hiltonol, are currently in clinical trials, and are very promising.

The drawback is that these studies do not usually rely on the research of this biomarker.

Generally ish,  naive T cells will express CD27, upregulate upon activation but lose when they are effector. Memory cells will go onto re-express CD27.

Ahlers, Blood, 2010 has a nice graphic associated with phenotype markers and their associations. Other good reading is from J. Borst who has done a lot of work looking at the CD27/CD70 pathway and how it relates to antigen specificity etc.

If you are talking about a gel retardation assay, or electrophoresis mobility shift assay, to detect DNA-protein interactions, we generally do not use either ethidium bromide or sytox green.  The DNA is usually labeled with 32-P because this is much more sensitive.  Also, intercalating DNA stains may interfere with the protein-DNA binding.

Dear Lihui,

Sterilization through 0.2 um should do the job, however, you may used glass ware or containers that were not sterilized.

The following is a procedure to get sterile pantothenate aliquot:

Pantothenic acid (FW 238.3 hemicalcium salt): Dissolve 0.405 g pantothenic acid in 100 ml water and filter sterilize to yield 17 mM (1000X) stock. Dispense 2 ml aliquots in sterile cryovials and store at -20°C or below; thawed stock can be kept at 4°C for up to 1 week.

Hoping this will be helpful,

Rafik

You might consider to convert your anti-NKG2A antibody of choice (and the respective isotype control) into Fab/F(ab)2-fragments - there are several commercial kits for that purpose that are easy to use and reliable. By using blocking Fab/F(ab)2-fragments you circumvent the potential problem of Fc-receptor-mediated activation. Good luck!

Elaine Cameron at the University of Manchester is working on a meta-synthesis of experiences of adherence to DMTs: http://www.crd.york.ac.uk/PROSPERO/display_record.asp?ID=CRD42015025254

Also: Johnson et al. (2006) perhaps? http://ijmsc.org/doi/abs/10.7224/1537-2073-8.1.11?=

Hi Dong Hyuk Joshua Seo,

I used salmonella typhimurium to check the macrophage phagocytic activity. It worked well for me. 

Best,

Biswaranjan Pradhan

Ph.D Student, SBS

NISER Bhubaneswar

Have you seen their web page? 

 Classically, the diagnosis

of syphilis is based on a combination of clinical history,

symptom presentation (if any), and serologic test results,

including non-treponemal and treponemal tests. Nontreponemal

tests (e.g., RPR, VDRL) are typically used

for screening, and measure a non-specific antibody to

treponemal infection. A reactive test can indicate recent

infection, but could be caused by other conditions unrelated

to syphilis. Treponemal tests (e.g., TPPA, TPHA, FTA-ABS,

EIA) detect antibody to syphilis and thus can confirm

exposure to treponemal disease. However, these tests cannot

distinguish venereal infection (syphilis) from non-venereal

treponemal diseases (e.g., yaws, pinta, bejel). Furthermore,

treponemal antibody persists for life, and thus treponemal

tests cannot distinguish between recent, active infection and

previously treated or old, non-contagious infection.

I agree with Dr. Ricardo

2015-cha-guidance-syphilis-testing-lac.pdf

Hi Leila!

good day!

I think the best bet for you to acquire the software is by communicating with the authors of this study:

Comparison of Parasite Burden Using Real-Time Polymerase Chain Reaction Assay and Limiting Dilution Assay in Leishmania major Infected Mouse

Somayeh GHOTLOO,1 Mostafa HAJI MOLLAHOSEINI,1,2 Ali NAJAFI,3 and Farshid YEGANEH1,2,*

Author information ► Article notes ► Copyright and License information ►

this is the link of this paper:

If I cannot access the information thru internet what I usually do is I just contact thru email the authors.

I wish you all the best in your research!

We may not be able to determine the geographic origin of the whole "species" Seratia marcesens but if there are specifc pathogenic variants of the species which have recently begun causing a disease in humans or plants, it could very well be possible to trace where the most recent source of this spread was.  This may cost many millions of dollars or much less, depending on many aspects of the "outbreak" and the natural range of the organism.   Some recent examples of this type of tracing are found by studying for example how the source of the enteropahtogenic E. coli (STEC serotype 0104H4-2011) which killed people in Europe in May-June 2011, was traced. 

Hi Corrado,

Here is a publication for the preservation of insects to be used in spectroscopy. This publication resulted from experience on field trips in Nigeria and Cameroon for insects that would be analyzed in Bamako, Mali. And Dr Floyd Dowell's experience in Tanzania. It is good you are not involving cryo-preservation at any point in the tropics.

I may suggest SILICA GEL for a short period. I had serious degradation of materials preserved in ethanol on one hand and on the other hand, methanol fixes molecules and may not yield very good results during DNA/RNA extraction.

I hope this helps

good luck

Lariam uses 1 tablet a week so you need to take 4 tablets.

Better to use. I used Lariam I didn't see side effects.

Zoya

Bill & Melinda Gates Foundation

Commonwealth Foundation

Elton John AIDS Foundation

European Union

Global Fund to Fight AIDS, Tuberculosis and Malaria

Hewlett Foundation

International HIV/AIDS Alliance

Government of the Netherlands

The Abbott Fund

US President’s Emergency Plan for AIDS Relief (PEPFAR)

UK Department for International Development (DFID)

UNAIDS

United States Agency for International Development (USAID)

United Nations Development Programme

World Health Organization

NACO, 9th Floor, Chanderlok Bldg., 36 Janpath,

New Delhi-110001 Tel: 23325337

Rajiv Gandhi Foundation

Rajendra Prasad Road

New Delhi - 110 001, India

Tel.: +91 11 2375 5117

Fax: +91 11 2375 5119

The Foundation supports HIV training programs for health professionals working with NGOs to treat the underprivileged throughout India. It also supports education, prevention and direct services programs for PLWHA as well as capacity-building grants for CBOs working in this area.

Vasavya Mahila Mandali

Benz Circle, Vijayawada - 520 010

Andhra Pradesh, India

Tel.: +91 86 6247 0966

Fax: +91 86 6247 3056

Vasavya is an intermediary for international and national

funders and governments. It supports home and community-based HIV/AIDS care and prevention programs.

MAMTA MAMTA

Health

Institute for Mother and Child

B-5, Greater Kailash Enclave-II

New Delhi, India 110048

Tel.: +91 11 2922 0210

Fax: +91 11 2922 0575

MAMTA is an intermediary for international and national funders and governments. It supports HIV/AIDS programs focusing on direct care, advocacy, and training throughout the country.

Palmyrah Workers Development Society (PWDS PWDS PWDS )

Crystal Street, Marthandam – 629165

Kanyakumari District

Tamil Nadu, India

Tel.: +91 46 5127 0241

Fax: +91 46 5127 0138

PWDS is an intermediary for international and national funders and governments. It supports CBOs and NGOs working with AIDS orphans as well as organizations providing community-based care and support programs for PLWHA.

LEPRA

Society

Post Box No. 1518

West Marredpally, Secunderabad

Andhra Pradesh, India

Tel.: +91 040 2780 2139

Fax: +91 040 2780 1391

LEPRA is an intermediary for international and national funders and governments. It supports a wide range of HIV/AIDS projects from direct care to advocacy to education and vocational programs throughout the country.

TEST Foundation

4, Sathalvar Street

Mugappair West

Chennai, India 600037

Tel.: +91 044 2624 4100

Fax: +91 044-2625 0315

TEST is both an operating foundation, running its own programs including home-based care and a hospice for AIDS patients, and a grantmaking

Yes, it sounds like the problem is with the titre method from the mouse lung as the animals are clearly getting sick. If you're avoiding freeze thawing after homogenization then the other factor that we've found affects titre a lot is the method of homogenization - we used to use hand-held "mixer" devices (with or without glass beads) and would get low titres, whereas the current Qiagen Tissue Lyser II machine gives us 100-fold higher titres from the same experimental set up. Whether this is because it's faster or simply macerates the tissue better (or both) I don't know.

Bottom line - if the animals are losing substantial amounts of weight, your system is basically working.

Paul

Hi Gaurav,

Thanks for the answer. In fact, I was aware of this system (a sort of hemodialysis) to remove endotoxin by polymyxin.  Rather I was curious to know about the fate of endotoxin trapped by its binder in vivo. If we consider reticuloendothelial system, it is likely to invoke innate immune system in somewhat altered manner (LPS in the endosomal compartment also detected by innate PRRs, until and unless it is degraded by the enzymatic system therein).

Dear Sally,

I am not sure I fully understand your question.

If you want do measure LPS level by LAL for a given sample.

You need to measure the dilution your sample you will need to fall in the middle of a standard curve (known quantities of LPS vs. OD read).

What I do not get is that helminth does not have LPS, so it will be always negative.

However if you which do Know how much helminth contamination you have in your sample.

Then I would go with a standard ELISA. On one site you do a standard curve with a knonw amount of helminth

Then the same with your sample in serial dilution and you see where you are in comparaison to the standard.

I hope its help.

Best,

Laurent

Dear Eric

There is an interesting paper you may be want to read:

Am J Pathol. 1988 Apr; 131(1): 48–52.

PMCID: PMC1880566

Immunoelectron microscopy of Schüffner's dots in Plasmodium vivax-infected human erythrocytes.

P. V. Udagama, C. T. Atkinson, J. S. Peiris, P. H. David, K. N. Mendis, and M. Aikawa

I attache the PDF.

Finally, please if you consider this answer appropriate, please upvote it using the green up arrow click. Thanks.

Dear Udoh, the study that has been done at Khartoum Sudan in tanneries may in one way correlate industries, chemicals, waste products and skin complications, hope you find some of the points you are looking for.

38. Kamal El-Din H El-Hassan, M.D, Yousif M El-Kordofani, MD, Bashir, A.H.H, MD. The Prevalence of Occupational Dermatosis among Workers in Khartoum State’s Tanneries. Original Article. AJDV, 2014, 3(2).

I think that this link may be useful for you.

Pathogenesis of human cytomegalovirus infection and ...

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de G Gerna - ‎2004 - ‎Cité 93 fois - ‎Autres articles

In acquired immunodeficiency syndrome patients with human cytomegalovirus (HCMV), disseminated infection, and end-organ disease, autopsy findings show ...

There is a nice matrix from CaptureSelect™ IgA Affinity Matrix (https://www.thermofisher.com/order/catalog/product/19428801L) able to bind all kind of IgA. I suggest to solubilize all the proteins ins the sputum first. For that, treat the sputum with N-acetylcysteine to break the mucus. Use 5 ml per gram of sputum. Add 10 volumes of PBS, incubate for 10 minutes. Centrifuge at 5000 g for 10 minutes and use the supernatant as binding material for IgA following the instruction of the manufacturer of the matrix. I hope this helps.

Hi Riva,

If you want to fix PMNs for ICC then fixed cells with 4 or 3.7% PFA at 4 degree Celsius for 30 minutes. Used zero acceleration and break for centrifugation when cells are live. Give minimum jerk to cells during resuspending pellet. Avoid using pipette for resuspending cells. Whenever you fix cells for microscopy then try to use the cells within 3-4 days after fixation.

This procedure works well with me. Try same.

Best luck!

Durgesh Pitale. 

There are no current classical antibodies to neutralize DENV, but there is a new development that shows promising results (antibodies against the E dimer conformation region, please refer to Galvin Screaton 2015), also there has been extensive research showing that the permissive cells in vivo are of stem cell which bear megakaryocyte phenotypes, this could also help understand dengue and develop strategies against it

Dear  Shankar Subramanian

It's possible that early initiation of antiretroviral treatment may expose to increasing viral resistance, and early treatment failures. We will have the answer to this concern when the results of ongoing studies in Zambia and South Africa will be published [1, 2].

It is also important to note that the major route of HIV transmission is sexual, more so in poorest areas. Immediate ART initiation thus favorably impacts not only the individual, but also all their sexual partners, providing tangible public health benefit.

The purpose of an early initiation of antiretroviral therapy is therefore not only improve the quality of life of people living with HIV but also to prevent HIV transmission. Thus, we can probably have a lower prevalence of HIV infection.

-------------------------

1. Iwuji CC, Orne-Gliemann J, Tanser F, et al. Evaluation of the impact of immediate versus WHO recommendations-guided antiretroviral therapy initiation on HIV incidence: the ANRS 12249 TasP (Treatment as Prevention) trial in Hlabisa sub-district, KwaZulu-Natal, South Africa: study protocol for a cluster randomised controlled trial. Trials 2013;14:230.

2. Hayes R, Ayles H, Beyers N, et al. HPTN 071 (PopART): rationale and design of a cluster-randomised trial of the population impact of an HIV combination prevention intervention including universal testing and treatment - a study protocol for a cluster randomised trial. Trials 2014;15:57

Dear Bhanu

I believe that the following review is very good for your it has a section about Plasmodium.

There are two approaches: The first, if you can't Perfuse your animals and the second if your can. If you cannot perfuse your animals (and can only fix them by submersion in formalin). One approach is to stain for different molecular wt proteins that exist in the sera. These proteins will leak across the BBB. For example you can immunocytochemically stain for IgG. ( While there are a lot of paper that use this approach, note: B cells found in the Brain may produce a confound with  this approach, so people also stain for other proteins as well). If you can perfuse, one approach is to follow the Saline perfusion with an HRP solution and then fix en-block with formalin. The you can detect the HRP with a DAB reaction.  This is just an explanation to get you started. You can see there are other alternatives to the Evan's blue.

I agree with dr Atef answer. It is important to exclude the presence of Pasteurella before starting experimental infection experiment. You may observe some respiratory signs if the animals are free of Pasteurella. If they are natural carrier of some strains and you use a laboratory strain  immune response  could be unsignifiant, since natural strains of Pasteurella may inhibit growing of laboratory strains

 Thanks for above answers. 

Yes, the level of LPS seems pretty high, especially when you look at this paper http://www.ncbi.nlm.nih.gov/pubmed/10515819 and normal donor have ~6pg/ml of LPS in plasma.

See the article below. Review on the subject

Pathogenesis of amoebic encephalitis: Are the amoebae being credited to an ‘inside job’ done by the host immune response?

Abstract

Pathogenic free living amoeba like Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris are known to cause fatal “amoebic meningoencephalitis” by acquiring different route of entries to the brain. The host immune response to these protist pathogens differs from each another, as evidenced by the postmortem gross and microscopic findings from the brains of the affected patients. Cited with the expression of ‘brain eating amoeba’ when the infection is caused by N. fowleri, this expression is making its way into parasitology journals and books. The impression that it imparts is, as if the brain damage is substantially due to the enzymes and toxins produced by this amoeba.

A detailed review of the literature, analysis of archived specimens and with our experimental assays, here we establish that with N. fowleri, Acanthamoeba and Balamuthia spp., the infections result in an extensive brain damage that in fact is substantially caused by the host immune response rather than the amoeba. Due to the comparatively larger sizes of these pathogens and the prior exposure of the amoebal antigen to the human body, the host immune system launches an amplified response that not only breaches the blood brain barrier (BBB), but also becomes the major cause of brain damage in Amoebic meningoencephalitis. It is our understanding that for N. fowleri the host immune response is dominated by acute inflammatory cytokines and that, in cases of Acanthamoeba and Balamuthia spp., it is the type IV hypersensitivity reaction that fundamentally not only contributes to disruption and leakiness of the blood brain barrier (BBB) but also causes the neuronal damage. The further intensification of brain damage is done by toxins and enzymes secreted by the amoeba, which causes the irreversible brain damage

The approach to the issue reported by Paul Digard and TimothyMiller is very interesting . It enters comparative immunology . it usually solves the equation with one unknown ( human weight of the animal to determine the dose to be injected ) It remains that when it comes to killed virus vaccine should increase the amount compared the live virus vaccine .

Thank you Amar! This indeed answer my question!!

Best,

Thibault

Thanks a lot! In case if someone interested I may forward my data set such that you may suggest me changes.

I agree with you. But I can not follow the third explanation. Do you mean the second challenge damage the immune tissue or organs? Such as, altering the total number of T cells or subgroups. I am reasrching the effect of  influenza vaccination on cognitive function during pregnancy. And i have found A(H1N1) influenza vaccine vaccination contribute to neurogenesis in offspring and pregnant mothers. I think the mechanism links to T cells in the periphery, such as the systemic T cells were recruited to the choroid plexes.

There are multiple reasons which can be summarised as under :

[a] Stage of liver cirrhosis - those with more advanced disease responds poorly.

[b] IL 28B typing - those with TT genotype fares poorer compared to CC type.

[c] Adherence to treatment - Those needing temporary drug withdrawal or dose modification due to adverse events have poorer outcome. 80% of total dose and 80% of total duration has been found to be effective. Ribavirin dose is more important.

[d] Some other factors which might have genetic basis are that males, presence of NAFLD and other chronic liver disease and coinfection with other hepatotropic viruses have poorer outcome. 

Note that because of the high number of T cells there will be competition for antigen, which will limit their response compared to an adoptive transfer model. Responses wiill also be less robust/comparable between animals (http://www.ncbi.nlm.nih.gov/pubmed/11163194). I  recommend for your assay that you stick with the adoptive transfer model.

Dear Dr. Hegazy,

The 2009 WHO guidelines entitled "WHO policy on Tuberculosis infection control in health-care facilities, congregate settings and households"  described all the different components on this topic  (http://www.who.int/tb/publications/2009/infection_control/en/). In addition, you can find specific recommendations related to building design in the following document: "TBCTA. IMPLEMENTING the WHO Policy on TB Infection Control in Health-Care Facilities, Congregate Settings and Households. A framework to plan, implement and scale-up TB infection control activities at country, facility and community level. November 2010. (http://www.tbcta.org)."

All the best,

Dr. Daniel Chemtob (Former WHO Medical Officer in charge at the Geneva WHO/HQ  of TB infection control worlwide)

Thank you prof. Amar Safdar.

I have already read this publication, we do not have the facilities  to run the biological testing, we are looking  for someone who can run the testing for us . 

Please try sending message to ROTACLONE manufacturers. They might be able to help.

You can, may be draw examples from the Indian model, although not completely integrated in its true sense currently. The National AIDS Control program nevertheless has worked progressively towards integration and now envisages to integrate the AIDS program completely in to the national health mission. Integration as I understand occurs at various levels and for various program components of the AIDS program. If your question is about the prevention and treatment component of HIV/AIDS then the answer would be yes it can be integrated one hundred percent. For example, testing for HIV can occur at any general laboratory, in most cases it would not require specialised training or equipment that is not manageable by a certified general laboratory technician. Now the larger problem I assume could be when we talk of the treatment component. If PLHIV have access to ARV medications provided by the government then it could be challenging, in terms of the capacities of the medical officers to provide standardized care based on the guidelines and also the laboratory support for periodic CD4 testing. Our experience in India is that integration is a slow drawn process especially for a concentrated epidemic such as ours. And, most off it depends on the existing public health infrastructure.

It is hard to find a regulated repository for serum samples from infected individuals as usually an approval from Ethics Committee is required for any specific purposes. It would be best if you find a collaborator in endemic areas.

Dying cells might exhibit low pH (acidic) due to the process of apoptosis or necrosis. TLR signal pathway is also relevant. 

From your question, you indicate that adherence to HIV and AIDS medicines in Nigeria is high and there are some factors that have been shown to contribute to that, i believe you want to establish which factors may be adapted into strategies for sustained high adherence.

Since you want to conduct a qualitative research, interview with the patients, health care providers, managers, caregivers and community organization representatives (such as support groups and home based care groups)  may be the way to go. You could use semi-structured questionnaires and focused group discussions and your questions should include some of the factors identified to result in high adherence and seek from the participants what they should be done. In your interviews you should have representation in terms of gender, age and if possible, duration on therapy. 

La Unidad de Babesiosis del Centro Nacional de Parasitología del INIFAP en Cuernavaca, México realiza tales diagnósticos y mediciones rutinariamente. Sin embargo juzgo muy difícil poder trabajar muestras de sueros de bovinos de Colombia, por las restricciones zoosanitarias. Indudablemente los colegas de EMBRAPA en Brasil ó del INTA en Argentina, son reconocidos internacionalmente y pueden apoyarlos. Igualmente, me imagino el Dr. Benavides que estuvo en CORPOICA muchos años y ahora se encuentra en la Universidad La Salle, podría ser una ayuda más próxima. Ánimo.

The only experience with this is some work with P. chabaudi. The main effect was that when we used pyrimethamine resistant parasites, they were definitely dependent on addition of 4-aminobenzoic acid (which was added to the drinking water for the mice) for a sustainable infection. With wild type parasites, this effect was not seen. My interpretation of that is that the mutation leading to resistance makes the folate production less efficient, and the addition will boost the folate synthesis enough to overcome this. 

Please refer to the following very good study and their Methods

If you have further questions, consider contacting the authors of this study

jgrosse4@jhmi.edu or dalmeid3@jhmi.edu.

To assess the bactericidal and sterilizing activity of the first-line regimen, we infected 223 female BALB/c mice by aerosol with M. tuberculosis strain H37Rv, achieving a mean implantation of 2.62 (SD 0.44) log 10 cfus in the lungs, with the spleens being culture-negative the day after infection.

Our objective was to establish a heavy infection with high cfu counts in the lungs and the spleens; as such, our original protocol was to initiate treatment 6 wk after infection. However, by 4 wk after infection, 12 mice had already died and all remaining mice were sick; therefore, we began treatment at 4 wk postinfection.

An additional 10 mice died during the first 4 d of treatment. To compensate for this loss of mice, we reduced the numbers of mice killed on day 0 (the day of treatment initiation) as well as at some of the relapse assessment time points (detailed in Materials and Methods).

We studied this adaptation through passaging in our previous paper (Ramadhany et al. 2012 ; Yasugi et al., 2012), when we cultured human influenza viruses in embryonated eggs, some mutations occurred in HA binding site even only after 1 time passaging. The population of viruses carrying mutation will become dominant through passaging in egg. To lessen these mutations, culturing human influenza viruses in MDCK cell might be better option.

Yes, in rare cases mycobacteria can be isolated from stool. But that does not indicate tuberculosis at all.

The diagnosis requires clinical evidence (unspecific abdominal and/or general signs, enlarged abdominal  lymph nodes, positive IGRA or Mendel-Mantoux test) and species diagnosis of the AFB´s by conventional or molecular (PCR) methods.

Personally, I think it is case dependent. First, it has to be determined whether its an unmasking or paradoxical IRIS. Starting steroids in under-controlled infections can be detrimental. 

There's a bunch of reasons.

Yes, most strains of flu (excepting HPAI H5 and H7s and a handful of plasminogen-dependent strains) are largely trypsin (or other exogenous protease) dependent for replication in cell culture. This is to cleave the HA so it becomes fusion competent (not receptor-binding competent - that activity is present in the uncleaved trimer). So if you're growing a stock or performing a low MOI multicycle replication experiment, serum is left out to avoid inactivating the trypsin. If you're doing a single cycle/high MOI infection and you don't care about titreing what comes out, then serum can be added back after the adsorption period.

Serum is left out at the point of infection because it contains inhibitors - decoy receptor molecules and other factors. There's a whole load of ancient literature on this (PMID 18904216 is one of the first) and if anyone's interested they can Pubmed "influenza", "serum" and "inhibitor" and read from the bottom up. The inhibition was virus strain and serum species specific, and although I don't recall the story in detail, I suspect the "francis inhibitor" and "beta inhibitors" etc were a mix of different flavours of sialic acid mucoids as well as other protein-based inhibitors that we might now call collectins/defensins etc.

Hi

dear doctor

Your answer is yes!

1. After 8 wk or more of gestation mother with evidence of immunity against varicella will transfer maternal IgG Ab to their fetus, thus their newborn infants already have positive Anti-varicella Ab that reflect passively-acquired maternal Ab (The IgG of the newborn is solely of maternal origin)

2. On the other hand, maternal Ab titers are affected  by her nutritional and immune

status,

3.In rare circumstances, mother may have low or absent levels of circulating IgG antibody. In these cases, the mother cannot transfer protection to her infant

4. You should check IG profile of mother in addition to serum IgG/IgM level of both infant and her mother simultaneously

 A quick high speed spin should help to get rid of the big pieces (RBC membranes, various agglutinated protein complexes).  You can also try modifying the elution buffer.  Since I don't know what is in your buffer here are  a few suggestions that might help

Adjust the Ca and Mg concentration down, maybe even to zero

tween or other gentle detergent that won't interfere with subsequent ELISA

mild reducing agents

quick high speed spin to get rid of the big pieces (RBC, protein complexes)

With all of these you will need to ensure they don't cause more problems than they solve

Hi! Concerning pH change, you have to be careful not to induce virus uncoating (if you want to obtain intact virions)...

Around 2 weeks after infection. 

IL-4 Attenuates Th1-Associated Chemokine Expression and Th1 Trafficking to Inflamed Tissues and Limits Pathogen Clearance

Christopher A. Lazarski,1 Jill Ford,1 Shoshana D. Katzman,1 Alexander F. Rosenberg,2 and Deborah J. Fowell1,*

The Centers for Disease Control and Prevention (CDC)- National Immunization Survey-Teen is a public domain survey conducted annually and may be helpful.  

Here is the link http://www.cdc.gov/nchs/nis/data_files_teen.htm

I am a doctoral student proposing a qualitative study of adolescent parents knowledge, attitudes, and beliefs about adolescent immunizations specifically HPV.  I am seeking qualitative questions used in other studies however running into problems getting permission to use an existing survey.  

Any suggestions.