What are Forbidden Cytokines?

Forbidden Cytokines and Autoimmunity

Chronic – long term immune stimulation drive distinct Th1 and Th2 cells to release ‘‘FORBIDDEN’’ cytokine secretions[viii]: secretion of Th2 cytokines from a Th1 T cells and vice versa. Such a phenomenon cannot be induced by classical antigenic stimulation. The immune cells become confused and respond inappropriately to immune messengers – producing confusing symptoms and responses to treatment.

T-cells secrete various cytokines through which they affect a broad spectrum of normal and pathological immune processes. Cytokine secretion by helper T cells is particularly important in autoimmunity[i] because chronic autoimmune diseases, such as Hashimoto’s Thyroiditis, Multiple Sclerosis, Type 1 Diabetes, and Rheumatoid Arthritis are predominantly caused by Th1 cells. Th2 cells can antagonize Th1 functions[ii] and in numerous autoimmune conditions prevent and/or cure autoimmune diseases.

All Components of the NEI Supersystem – including Cytokines and Chemokines are Dose-Dependent

However, recent studies found exceptions to this rule, suggesting that the behavior of a given T cell population may be unpredictable in its cytokine secretion profile. For example, (i) Th2-type T cells can be not only inefficient suppressors of autoimmune conditions induced by Th1 cells,[iii] but can cause Autoimmune conditions;[iv] thus Th1 and Th2 cells can both promote autoimmune conditions; (ii) Th0-type T cells (producing both Th1 and Th2 cytokines) can stimulate autoimmune conditions and are able to instigate Autoimmune conditions;[v] and (iii) Th2-type T cells can induce pancreatitis and diabetes in immune-compromised nonobese individuals with blood sugar problems.[vi]

The cytokine secretion of the same T cell population is different in the lymph nodes (producing both Th1 and Th2 cytokines) than in the central nervous system (CNS) environment (producing only Th1 cytokines). This observation suggests that within the CNS, specific factors (mainly IL-12 producing microglia acting as APCs, not neurons) can modulate the cytokine secretion of Tcells, can select Th1/Th2 pathway, and can control effector CD4+ T cell cytokine profile in Autoimmune conditions.[vii]

Four neuropeptides (NPs): somatostatin, calcitonin gene-related peptide, neuropeptide Y, and substance P, in the absence of any additional factors, directly induce a increased secretion of cytokines [interleukin 2 (IL-2), interferon-g, IL-4, and IL-10) from T cells. Furthermore, these NPs drive distinct Th1 and Th2 populations to a ‘‘FORBIDDEN’’ cytokine secretion[viii]: secretion of Th2 cytokines from a Th1 T cell line and vice versa. Such a phenomenon cannot be induced by classical antigenic stimulation.

The nervous system, through these NPs interacting with their specific T cell-expressed receptors, can lead to the secretion of both typical and atypical cytokines, leading to the breakdown of the commitment to a distinct Th phenotype, and a potentially altered function and destiny of T cells in vivo.

Nerve fibers that release NPs are widespread in the mammalian central and peripheral nervous systems, in certain endocrine tissues, and in all the primary and secondary lymphoid organs.[ix]

Inflammation Is An Immune Response

Inflammation is part of the non-specific immune response that occurs in reaction to any type of bodily injury. In some disorders, the inflammatory process − which under normal conditions is self-limiting − becomes continuous and chronic inflammatory diseases might develop subsequently.

How do they separate the Immune Response from Inflammation?

Today, very few health practitioners could tell you that inflammation is part of the nonspecific immune response that occurs in reaction to any type of bodily injury and that the cardinal signs of inflammation can be explained by increased blood flow, elevated cellular metabolism, vasodilatation, release of soluble mediators, extravasation of fluids and cellular influx. They would never associate inflammation with an immune response. An immune response in their world would be something that occurs only with bacteria or viruses.

Inflammation is part of the complex biological response of body tissues to harmful stimuli, such as pathogens, damaged cells, or irritants,[1] and is a protective response involving immune cells, blood vessels, and cytokine / chemokine immune mediators. The function of inflammation is to eliminate the initial cause of cell injury, clear out dead or damaged cells and tissues damaged from the original insult and the inflammatory process, and initiate tissue repair.

Acute (Short-term) or Chronic (Long-term) Inflammation

Inflammation can be classified as either acute or chronic. Acute inflammation is the initial response of the body to harmful stimuli and is achieved by the increased movement of plasma and leukocytes (especially granulocytes) from the blood into the injured tissues. A series of biochemical events propagates and matures the inflammatory response, involving the local vascular system, the immune system, and various cells within the injured tissue. Prolonged inflammation, known as chronic inflammation, leads to a progressive shift in the type of cells present at the site of inflammation, such as mononuclear cells, and is characterized by simultaneous destruction and healing of the tissue from the inflammatory process. Over time the destruction will become greater than the healing of the tissue.

Continuous Inflammation = Continuous Immune Response

In some disorders the inflammatory process, which under normal conditions is self-limiting, becomes continuous and chronic inflammatory diseases develop subsequently. This inflammatory process is enhanced by the over-use of immune stimulating supplements, probiotics and bone broth.

Is the peer group always correct? You are not allowed to think differently from the group.

Chronic inflammation culminates in devastating events that can lead ultimately to major organ dysfunction due to abnormalities in tissue architecture after regeneration and replacement by non-functional fibrous tissue. Once this stage is reached, it becomes more difficult to reverse, it at all.

Inflammation is not a synonym for infection. Infection describes the interaction between the action of microbial invasion and the reaction of the body’s inflammatory response — the two components are considered together when discussing an infection, and the word is used to imply a microbial invasive cause for the observed inflammatory reaction. Inflammation on the other hand describes purely the body’s immunovascular response, whatever the cause may be.

Therapies developed against these illnesses are not foolproof, as when more than one factor comes into play, the number of interactions also raises exponentially.  We might well be standing before a family of very closely related diseases with ubiquitous symptoms common to all. Yet differing only due to very small changes at some point along the inflammatory cascade. The common denominator in these conditions is the immune system.

And so these Doctors Disputed loud and long Each in his own opinion Exceeding stiff and strong, Though each was partly in the right, And all were in the wrong! The Blind Men and the Elephant: 1865

Understanding the cell-signalling networks involved in chronic inflammation gives rise to new insight into how drugs target the immune system to suppress troublesome diseases, e.g. ‘old’ therapies such as glucocorticoid
and immunosuppressants work by suppressing the Th2 responses.

[1] Ferrero-Miliani L, Nielsen OH, Andersen PS, Girardin SE; Nielsen; Andersen; Girardin (February 2007). “Chronic inflammation: importance of NOD2 and NALP3 in interleukin-1beta generation”. Clin. Exp. Immunol. 147

Are You Eating Healthy?

Growers Lie About Being Resellers Of Produce

The deceptive “Growers” can charge more if people think it was grown locally.

I keep telling people to ask if the seller grows the produce at the farmers market. Can you visit their farm where the produce is grown? The trouble is people are so disconnected and familiar with seeing produce at the grocery store year around. They have no idea what is being harvested or where it is grown.

They are more emotionally connected to the idea of buying their “organic” produce from a local grower while spending some outdoor time at the “Farmers Market”, than finding out where the “organic” produce is actually procured from.

I stopped going years ago when I asked and found out the “Growers” at the Farmer’s Market were buying the seconds and blemished produce the major grocery store chains rejected.

This is what happens when you grow up on a Dairy farm, helping to grow a garden to feed a family of eight. Oh, by the way, don’t walk by the garden without pulling some weeds. You learn what is in season and when it is harvested during the growing season.

California Blend Veggies From China At Whole Foods?

Even Whole Foods is not selling what people think they are selling. Granted this video is from 2008. I seriously doubt much has changed.

Living the Nocebo Effect

Disease Diagnosis are not Entities entering your body seeking to do you harm.

And so these Doctors Disputed Loud and Long, Each in his own opinion, Exceeding stiff and strong, Though each was partly in the right, And all were in the wrong! The Blind Men and the Elephant: 1865

Too many people convinced of the “Nocebo Effect” by Professional and Social Media Influencers are unable to see or experience any improvement.

What does “Better” mean to you? Are you on a never-ending-treatment-protocol? What happens if you stop it? If you get worse – You are not “Better”. You are dependent.

Define Better! What Does “BETTER” Mean To You?

Too many times patients get to a point of having to decide whether to be sick or no longer need treatment, decide to stay sick. Mostly due to their Social Media Support Group pulling them back into sickness.

Too many people are convinced by Professional or Social Media Influencers of the futility of health and the inevitability of sickness to support sales of their effectively-ineffective products and treatments.

ReThink Candida

From the time you collected the culture Stool sample at your home. Candida Continues to grow without any competition because 95% of the original microbes in the stool sample have died or become dormant while being transported to the laboratory. Lacking any competition for food, Candida continues to proliferate during the incubation at the Laboratory. This overgrowth during transportation and preparation fuels the common belief of many Doctors and People alike of the deleterious health effects of Candida overgrowth in the body. This belief has resulted in the inappropriate use of Candida drugs, supplements and diets. Stool Culture laboratory results are likely erroneous and can lead to inappropriate patient treatment for Candida, Yeast and Mold.

Candida lacking any competition out competes 95% of the original dead or dormant microbes in the stool sample.

Documented Limitation of Culture Stool Assessment

Introduction

In the last decade, DNA sequencing has played a pivotal role in our knowledge on microbial diversity providing an outstanding tool for the detection, identification, and characterization of microorganisms.(1,2) DNA assessment has greater efficiency, reliability, and reproducibility, as well as providing both qualitative and quantitative data. DNA analysis is the new gold standard for identification of bacteria in clinical microbiology and has greatly facilitated the identification and classification of intestinal microbiota composition.(3-7) Much of this has been done with techniques based on 16S rDNA gene, which include fluorescent in situ hybridization, denaturing gradient gel electrophoresis, and temperature gradient gel electrophoresis.(4,8-10) 

DNA analysis is the new gold standard for identification of bacteria in clinical microbiology and has greatly facilitated the identification and classification of intestinal microbiota composition.3-7

Richard Lord

So why does anyone use the culture method? Conventional bacteriological methods, such as microscopy and culture, have been used for analysis and/or quantification of the intestinal microbiota for decades.(8-10) Barriers, such as investment in new equipment and staff, as well as commitments to on-going analysis and research collaborations, interfere with what many laboratories are able to offer. Translating science to accepted standard clinical protocol takes time.(11) Culture is still readily available and reasonably accurate, so clinicians continue to use it even though there are known significant limitations which include: transport issues, indeterminate and presumptive quantification and identification, as well as subjectively demanding techniques., In this review, we discuss six major pitfalls of conventional stool assessment. 

1. Transport Issues

The primary issue with culture analysis is that of transport. Since analysis is culture dependent, sample collection must be done using nutrient broth containers to keep as many bacteria alive as possible in transport. This broth allows continued growth of those species most satisfied with the broth medium during transport, until the sample is finally plated for culture. This growth results in a significant change in the balance of microbes present since some species will more actively grow at the expense of others. DNA analysis eliminates this problem by placing the specimen in Formalin vials for transport. Formalin immediately kills all organisms, freezing the exact balance present at the time of collection. Since DNA identification is only looking for the genes of the microbiota, living specimens are not necessary. This technique allows the clinician to develop the most appropriate therapy based on the patient’s true gut microbiota, resulting in better clinical results.

Recent studies in our laboratory illustrate this situation. One specimen was placed in two vials: one vial containing formalin and another a nutrient broth commonly used for transport. Both vials were incubated for three days at room temperature, then DNA was extracted. The extract was incubated with three different restriction enzymes which cleave the DNA at specific base pair sequences (Figure 1).

The digestate was amplified by polymerase chain reaction (PCR), and then placed on an agarose gel plate and the DNA fragments were electrophoretically separated. Since restriction enzymes cleave the DNA only at specific base pair sequences, broths with identical populations of microbes would produce the same patterns of banding in the electrophoretic runs. If, however, there were different amounts and types of microbiota in the two tubes, then differences would appear in the banding patterns. This was clearly demonstrated in the experiment (Figure 2 ).

Figure 2. Three restriction enzymes HAEIII, Mbol, and Sau3A digest the same two samples. One sample comes from culture and sensitivity (CS) vial containing a nutrient broth. The other sample vial contains formalin (F). Both vials have incubated for three days. The banding patterns are significantly different for the two vials indicating the populations of microbiota present are significantly different. In addition, note the CS vials have much fewer and heavier bands indicating that the microbial populations have reduced diversity due to the anaerobic die off and those viable ones have overgrown. Some bands present in the F vials are not detected in the CS vials where whole groups have died off. L is a calibration of known DNA sizes.

Another observation from the data is the loss of banding in the nutrient broth vial over time, indicating overgrowth of aerobes at the expense of the anaerobic populations and consequent loss of diversity. Opportunistic, potentially pathogenic organisms can also overgrow under these conditions. This overgrowth has been seen in our laboratory. For these reasons, nutrient broth transport vials cannot provide the specimen integrity required for accurate measurement of gut microbes.

Figures 3 through 5 illustrate the growth response of Bifidobacteria, Candida species and Staphylococcus aureus in one subject over three days. This data indicates that clinical laboratory data relying on transport of the specimen to the laboratory for culture are likely erroneous and can lead to inappropriate patient treatment. Candida species overgrowth is particularly notable (Figure 5). As these data indicate, Candida proliferates in nutrient broth transport media. This overgrowth in transport has likely fueled the common belief of many practitioners of the deleterious health effects of Candida albicans overgrowth in the gut. This belief may have resulted in inappropriate prescription of anti-fungal agents.

Transport Effects on Bifidobacteria Concentrations
Transport Effects on Staphylococcus aureus.
Transport Effects on Candida Species

This data indicates that clinical laboratory data relying on transport of the specimen to the laboratory for culture are likely erroneous and can lead to inappropriate patient treatment.

Richard Lord

2. Quantitative Assessment

In the Metametrix DNA assessment, quantification of predominate, opportunistic, and pathogenic bacteria are given as exact amounts. Yeast/fungal values are given as a plus 1, 2, 3, or 4 and represent a specific value range. For example, a +1 is specifically quantified to mean 100 – 999 pica grams of DNA per gram with asystematic reproducible result. Similarly, bacterial assessments are quantitatively reported as Colony Forming Units per gram sample.

Stool Culture Assessment is a “Best Guess”, subject to “Human Error”.

In laboratories that utilize culture assessments, values of quantity are significantly less specific or reproducible. For example, when samples come into the laboratory, they are plated onto a dish with selective or differential media. The technician dips a tool, generally a cotton tipped applicator, into the sample and puts it on a plate that is divided into four quadrants. The technician starts streaking in quadrant 1, then drags the tool to quadrant 2, 3, then 4 in sequence. The dishes are incubated overnight at which time they are assessed. If the bacteria grew in the first quadrant only, it is called a +1. The assumption is that there was only enough of the bacteria in the sample, due to low concentrations, to grow only in the first quadrant. The first quadrant is expected to have the greatest numbers of bacteria since that is where the streaking starts. If colonies appear in all four quadrants, it is called a +4. The fourth quadrant would be expected to have the least amount of original bacteria present since it is the last streaked. Thus, if something grew in the fourth quadrant, it is assumed that it grew there because bacteria were present in high concentrations in the sample. No procedure or assessment is done to confirm this assumption. No standardization for original amount of sample on the tool exists. A +4 theoretically means that there was enough of the bacteria on the sample that it stayed on the tool while it was dragged from the first to the fourth quadrant. However, with no standardization, these steps are at best a subjective judgment by the technician.

Additionally, if growth is found in the first and fourth quadrant, but not in the second and third, some technicians may call this a +4, while another may call it a +2. These results are technician based, and allow for human error, such as the technician turning the tool differently in the last quadrant. Consequently, a +1, +2, +3, or +4 from culture is an unverified guess based on where the microbe grew in the culture dish. This is at best a qualitative assessment of bacterial presence in the sample. These discrepancies are some of the reasons hospital-based laboratories give results as positive or negative and do not attempt to quantify when using culture techniques.

3. Presumptive Diagnosis/Identification

To identify individual microbes growing on the initial plates, individual colonies are selected and grown out again. Using a process of elimination composed of several if-then steps, a series of tests are done which yield a diagnosis of the most likely organism present. Typical characteristics such as colony morphology, key biochemical reactions, and drug susceptibility patterns assist in establishing the suppositious identification of the microorganism. However, most labs imply that this diagnostic identification of the microbe is definitive. It is not definitive, but at best gives a percentage ‘best estimate’  of the likelihood of identification accuracy.

It is widely accepted that plate culturing techniques reveal only a small portion of the true microbial population, primarily due to an inability of detecting organisms that might not be cultivable with known existing media, temperature specifics, phases of metabolic activity, and the inability of recovering known microorganisms which are viable but enter a non-cultivable state.(13-15) DNA analysis became the standard of research more than a decade ago, partially due to the subjective nature of culture ID systems. The sensitivity of conventional culture method was compared to DNA analysis for identifying Shigella spp. and enteroinvasive Escherichia coli (EIEC) in known cases of dysentery. DNA had an identification accuracy rate of 96%, compared to culture’s identification rate of 54%.(12) 

However, most labs imply that this diagnostic identification of the microbe is definitive. It is not definitive, but at best gives a percentage ‘best estimate’ of the likelihood of identification accuracy. 

Richard Lord

4. Identification of Predominate Bacteria

Bifidobacteria are among the first to establish themselves in the infant’s intestinal tract, and in the healthy infant, they predominate. While it would be impossible under normal life circumstances to have 0+ as a level of Bifidobacteria, that level is often reported in laboratories utilizing culture techniques. In these laboratories, Lactobacillus and Bifidobacteria, both facultative anaerobes, are identified primarily based on selective growth media. Though further enzyme tests such as coagulase or catalase may be done to hone the identification of Lactobacillus, Bifidobacteria identification is done solely on this single growth. Quantification is also made from the growth on these plates, as described above. These phenotypic methods suffer from a lack of reproducibility generated by conditions of culture used in different laboratories and to the diversity of strains that comprise the recognized species. DNA identification and quantification of Lactobacillus and Bifidobacteria is exact.

Doctor recommended the Best Probiotic “Ever”. Why did the PCR test show little colonization of Bifidobacter?
LooK at the six month cost of the “Best Probiotic Ever” with nothing to show for it.

5. Parasitology

Parasitology is yet another field of microbiology to be greatly improved by molecular technologies. Classically, parasites have been identified by microscopy and enzyme immunoassays.(17) In recent studies, DNA techniques have proven to be more sensitive and specific than classic laboratory methods.(4,17,18) One example is Giardia. Since Giardia cysts are shed sporadically and the number may vary from day to day, laboratories have adopted multiple stool collections to help increase identification rates for all parasite examinations.(4) Even with the advent of antigen detection systems, there has long been uncertainty in diagnosis when no ova or parasites are found. With PCR technology, only one fecal sample is needed for 100% sensitivity and specificity in parasitology examinations. The clinician needs to be aware that assessment of parasites, while good by microscopy, is much more accurate with DNA based technologies.

Tape Worm put into sample container. “Optimized Parasite Recovery” by lab found no parasites.

6. Automated Identification Systems

There are three main automated systems in use for microbial identification: the MicroScan, VITEK 2, and Crystal GP. All use the if-then sequences discussed earlier to identify characteristics of microbiota and produce proposed bacterial identification with a given accuracy expressed as a percentage.

Vitek, Vitek 2, Maldi-TOF and Crystal GP all require “CULTURABLE” microbes for identification.

The accuracy of these systems was evaluated by identifying coagulase-negative staphylococci (CNS). Verification was done with DNA by 16S rRNA sequencing. The MicroScan, VITEK 2, and Crystal GP systems correctly identified 82.5%, 87.5%, and 67.5% of the isolates, respectively. Misidentification was the main problem in MicroScan and Crystal GP systems, whereas the main problem of VITEK 2 was low-level discrimination. None of the three phenotypic systems tested could accurately and reliably identify CNS at the species level as well as the 16S rRNA technique.19 There are two types of the VITEK systems, the VITEK 2 fluorescent card and the VITEK 2 colorimetric card assay. In another study, the accuracy of identification at the species level for the VITEK 2 fluorescent card, the VITEK 2 colorimetric card assays, and the API 20 NE (another identification system), were 48%, 59%, and 46%, respectively. In comparing 16S rRNA gene sequencing to the VITEK 2 fluorescent card, the VITEK 2 only identified 43% of the isolates correctly at the species level, a low rate., Due to this low rate of identification, the authors proposed an algorithm for proper identification of non-fermenting gram-negative rods. In the algorithm, isolates with only good or acceptable identification to species level should be subjected to 16S rRNA gene sequencing when accurate species assignment is sought.  

Conclusion

DNA assessment is specific and accurate in the identification and quantification of fecal microbiota. As discussed above, culture methods are relatively inefficient, time consuming, and labor intensive, often leaving diagnosis obscure. DNA is highly sensitive and specific. Beginning with the sample collection, DNA assessment is superior because it avoids the pitfalls of transport that culture methods suffer from and allows the identification of true gut microbiota, as well as the anaerobic population comprising 95% of gut microbiota. The results are the actual types and amounts at the time of collection, not what survived in a growth medium. DNA further reports the microbiota as true numbers, values that were specifically quantified, not an assumption. The ability of DNA to accurately identify microbiota has been one of the primary reasons why it has been so embraced by the research community for the last decade. Research on Bifidobacteria and Lactobacillus has significantly expanded since the introduction of DNA analysis. DNA techniques are known to be more sensitive and specific than classic laboratory methods.,, Verification of the accuracy of culture assessment tools and equipment is done by comparing their results to the “actual” value, which is determined by DNA assessments.

Though culture has been correlated to clinical features in older research studies, we now know much of the data was incorrect.

Richard Lord

Sellers of conventional stool tests argue that DNA assessment does not have the years of clinical background that culture has. This point of course is misleading. Though culture has been correlated to clinical features in older research studies, we now know much of the data was incorrect, due to the significant limitations of culture and the indeterminate results it produces. Researchers have been actively reevaluating past research for this reason. Specific values of gut bacteria have been identified for normal healthy populations using DNA assessments, as well as for populations of selected groups and specific disease conditions., 

A notable example many clinicians are now aware of is the research of Dr. Gordon and Dr. Ley who discovered that obese volunteers had greater amounts of Firmicutes and significantly less Bacteroidetes than the lean participants., In their research on fat bugs, they performed a comparative 16S-rRNA-gene sequence-based survey (DNA analysis) of the distal gut microbiota – NOT CULTURE. Gordon’s team could not and would not have done this research with culture, because it cannot produce this information. DNA analysis is the most thorough and reliable assessment of the gut microbiota. It eliminates the major limitations of conventional stool testing using culture methods and offers advanced knowledge for more effective patient treatment.

References

  1. Woo PC, Lau SK, Teng JL, Tse H, Yuen KY. Then and now: use of 16S rDNA gene sequencing for bacterial identification and discovery of novel bacteria in clinical microbiology laboratories. Clin Microbiol Infect. Oct 2008;14(10):908-934. 
  2. Woese CR. Bacterial evolution. Microbiol Rev. Jun 1987;51(2):221-271.
  3. Clarridge JE, 3rd. Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev. Oct 2004;17(4):840-862, table of contents.
  4. Ghosh S, Debnath A, Sil A, De S, Chattopadhyay DJ, Das P. PCR detection of Giardia lamblia in stool: targeting intergenic spacer region of multicopy rRNA gene. Mol Cell Probes. Jun 2000;14(3):181-189.
  5. Langendijk PS, Schut F, Jansen GJ, et al. Quantitative fluorescence in situ hybridization of Bifidobacterium spp. with genus-specific 16S rRNA-targeted probes and its application in fecal samples. Apple Environ Microbiol. Aug 1995;61(8):3069-3075.
  6. Welling GW, Elfferich P, Raangs GC, Wildeboer-Veloo AC, Jansen GJ, Degener JE. 16S ribosomal RNA-targeted oligonucleotide probes for monitoring of intestinal tract bacteria. Scand J Gastroenterol Suppl. 1997;222:17-19. 
  7. Delgado S, Suarez A, Mayo B. Identification of dominant bacteria in feces and colonic mucosa from healthy Spanish adults by culturing and by 16S rDNA sequence analysis. Dig Dis Sci. Apr 2006;51(4):744-751. 
  8. Finegold S, Sutter V, Mathisen G. Normal indigenous intestinal flora. New York: Academic Press; 1983. 
  9. O’Sullivan DJ. Methods of analysis of the intestinal microflora. In: Tannock GW, ed. Probiotics: a critical review. Wymondham: Horizon Scientific Press; 1999:23-44. 
  10. Tannock GW. Analysis of the intestinal microflora: a renaissance. Antonie Van Leeuwenhoek. Jul-Nov 1999;76(1-4):265-278.
  11. NIH. Translational Research – NIH Roadmap for Medical Research. http://nihroadmap.nih.gov/clinicalresearch/overview-translational.asp. March 19, 2008. 
  12. Dutta S, Chatterjee A, Dutta P, et al. Sensitivity and performance characteristics of a direct PCR with stool samples in comparison to conventional techniques for diagnosis of Shigella and enteroinvasive Escherichia coli infection in children with acute diarrhoea in Calcutta, India. J Med Microbiol. Aug 2001;50(8):667- 674. 
  13. Lleo MM, Bonato B, Tafi MC, Signoretto C, Pruzzo C, Canepari P. Molecular vs culture methods for the detection of bacterial faecal indicators in groundwater for human use. Lett Appl Microbiol. 2005;40(4):289-294.
  14. del Mar Lleo M, Tafi MC, Signoretto C, Dal Cero C, Canepari P. Competitive polymerase chain reaction for quantification of nonculturable Enterococcus faecalis cells in lake water. FEMS Microbiol Ecol Dec 1 1999;30(4):345-353.
  15. Lleo MM, Tafi MC, Canepari P. Nonculturable Enterococcus faecalis cells are metabolically active and capable of resuming active growth. Syst Appl Microbiol. Aug 1998;21(3):333-339. 
  16. Ballongue J. Bifidobacteria and probiotic action. Vol pages 357-428. NewYork: Marcel Dekker Inc.; 1993.
  17. Verweij JJ, Blange RA, Templeton K, et al. Simultaneous detection of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum in fecal samples by using multiplex real-time PCR. J Clin Microbiol. Mar 2004;42(3):1220-1223.
  18. Morgan UM, Pallant L, Dwyer BW, Forbes DA, Rich G, Thompson RC. Comparison of PCR and microscopy for detection of Cryptosporidium parvum in human fecal specimens: clinical trial. J Clin Microbiol. Apr 1998;36(4):995-998.
  19. Kim M, Heo SR, Choi SH, et al. Comparison of the MicroScan, VITEK 2, and Crystal GP with 16S rRNA sequencing and MicroSeq 500 v2.0 analysis for coagulase-negative Staphylococci. BMC Microbiol. 2008;8:233.
  20. Ling TK, Liu ZK, Cheng AF. Evaluation of the VITEK 2 system for rapid direct identification and susceptibility testing of gram negative bacilli from positive blood cultures. J Clin Microbiol. Oct 2003;41(10):4705-4707.
  21. Zbinden A, Bottger EC, Bosshard PP, Zbinden R. Evaluation of the colorimetric VITEK 2 card for identification of gram-negative nonfermentative rods: comparison to 16S rRNA gene sequencing. J Clin Microbiol. Jul 2007;45(7):2270-2273.
  22. Bosshard PP, Zbinden R, Abels S, Boddinghaus B, Altwegg M, Bottger EC. 16S rRNA gene sequencing versus the API 20 NE system and the VITEK 2 ID-GNB card for identification of nonfermenting Gram-negative bacteria in the clinical laboratory. J Clin Microbiol. Apr 2006;44(4):1359-1366.
  23. Hayashi H, Sakamoto M, Kitahara M, Benno Y. Molecular analysis of fecal microbiota in elderly individuals using 16S rDNA library and T-RFLP. Microbiol Immunol. 2003;47(8):557-570.
  24. Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Microbial ecology: human gut microbes associated with obesity. Nature. Dec 21 2006;444(7122):1022-1023.
  25. Turnbaugh PJ, Hamady M, Yatsunenko T, et al. A core gut microbiome in obese and lean twins. Nature. Jan 22 2009;457(7228):480-484.

I have a Positive Antibody Test Result?

Do you have Positive Antibody Test Results? Are you questioning the results? Are you feeling no different after months on your treatment regimen? Antibody testing is not as infallible as they are made out to be.

If the results of Antibody tests are tied only to symptoms, you are guaranteed to be diagnosed with whatever condition you are being tested for.

Fallibility of Antibody Testing

I first began to question antibody testing after working with a 16 year old cheerleader that would go violently manic after eating gluten. I am talking about call 911 – violently manic. How could that occur where the behavior and symptoms do not match the lab test?

16 year old Cheerleader would go violently manic after eating gluten. Serum (blood) test reported Zero IgA Gluten antibodies. Stool test IgA Gluten antibodies was 5.3 – equivocal. The IgAs are the same throughout the body. The problem is not in the lab tests. The problem lies in her immune systems inappropriate response.

My personal lab tests were the opposite of the Cheerleaders. My saliva IgA test results were positive. My stool IgA results showed my intestinal tract was producing very low quantities of IgAs.

My personal Gluten/gliadin IgA test results done a week apart. I experienced severe brain fog and dyslexia after eating gluten. That was then. Now am able to eat gluten without any symptoms.

My first thoughts were – maybe the saliva, serum and gut produce different types of IgAs. Not the case. IgAs are the same through out the body. My next question is how could be IgAs produced in different parts of the body be so vastly different in the body?

I spoke with the Professional Influencers, who promote Antibody testing. Their answers were to blame the lab tests. Okay??? If the lab tests are a problem. Why are you still using and promoting them in your classes? Foot steps. Click – phone disconnected.

Dogma of Antibody Testing.

Immune Responses cause Inflammation.
Abnormal or Positive Antibody results does not indicate an active Immune Response.

The current dogma is that Immune cells are stimulated by Antibodies. The Antibody tests are thought to be absolutely accurate and infallible. They believe as long as antibodies are present, the antibodies are constantly stimulating an immune response. They also are lead to believe the presence of antibodies are from an ongoing infection. Typically, IgG antibodies are produced in the first phase of immune responses. However, the immune response to IgA, IgG, and IgM declines or is terminated along the course of the disease in most patients. IgM antibodies are indicative of an ongoing immune response., This suggests that the antibody response eventually burns out. Studies have shown the detection of IgA and IgM antibodies in 62% and 61% of IgG antibodies have been found in autoimmune patients after 7 years.

36 year old woman told by MD, she needed MMR booster shot because her children are starting school.

This suggests that the antibody response eventually burns out.

The presence of TPO, TAA and Gluten antibodies does not indicate an active Hashimoto’s – Thyroid Immune Attack.

A resolution of the disease is not associated with a decrease in antibodies. Only those with a short disease duration are likely to experience a decrease in antibodies levels. For most autoimmune patients there is no significant change in antibody levels, However, patients with immune calming/quieting treatment, experienced a 20% decline in their antibody levels. Normalization in the levels of antibodies with treatment intervention occurs in less that 7% of patients. The success of clinical interventions in lowering antibody levels (and thus likely minimizing the pathogenic effects of autoantibody binding) appears to be dependent on disease duration. The association of shorter disease duration with greater declines in antibody levels is highly consistent with the growing body of evidence that shows improved clinical outcomes with earlier disease intervention in autoimmunity.,,

36 year old woman told by MD, she needed MMR booster shot because her children are starting school.

Approximately one-half of patients with Rheumatoid Arthritis had positive IgM-RF and/or antibodies on at least 1 occasion, almost 5 years prior to disease onset. More than 50% of autoimmune patients may have positive antibodies upwards of 13 years after disease resolution, regardless of treatment.

Do You Need More Examples Of Questionable Antibody Tests?

Most Doctors will blame the Laboratory Company. It is not the Laboratory Company. They are reporting accurate results for what they are testing. It is your “Immune System” that is the problem. Your Immune System is not just compromised. It is broken.

36 year old woman told by MD, she needed MMR booster shot because her children are starting school.

How many $$$thousands of Dollars are spent on Antibody testing. One patient that came to my office for help spent in excess of $4,500 in antibody lab testing. All of the results were “IN RANGE” negative.

MRT Cytokine test shows severe reactivity to yeast while Cyrex antibody test shows none.

Until the Immune System is calmed and quieted, antibody tests are pretty useless, confusing and frustrating. People are constantly changing their diet to comply with their food allergy test. Many tests results are over five years old. Many people have done multiple food allergy tests. Yet, the person is still attempting to comply dietarily with all of the tests results, while sliding down a slippery slope and nothing it working. So, they seek out another Diagnosis du jour. Never letting go of the old diagnosis or test result and incorporating the new.

The Immune System is dynamic and responses to food can change over time

Has anyone ever stopped to consider the Immune System is dynamic and responses to food can change over time? Food sensitivities/allergies are not permanent, forever and ever, written in stone.

MRT Cytokine test shows severe reactivity to millet, oats and rice while Cyrex antibody test shows none.

Whatever food you frequently consume to comply with avoiding your food sensitivity/allergy meals will become the new food sensitivity/allergy.

MRT Cytokine test shows reactivity to milk and dairy while Cyrex antibody test shows none.

I have not used a food allergy/sensitivity test for about 15 years. It is unlikely that I would recommend them as long as people are bringing these conflicting test results.

As a farmer, I question how tomatoes and other summer vegetables are “Fresh” when there is snow on the ground.

It is not the foods that are causing the problem. Except for lectins. Lectins are a problem. People are “eating only fresh”, “eating organic” foods that have been harvest when they were not ripe, gassed to prevent further ripening, stored in Atmosphere Controlled Storage for months or years, shipped to the wholesaler, gassed again to restart ripening, then put out in the “fresh produce” aisle. Yes, even at Whole Foods and Trader Joes. I know how you think!!!

Atmosphere Controlled Storage of “Fresh” produce allows them to be stored for months or years before being sold as “Fresh” produce.

I focus instead on restoring the production and sequencing of digestive chemistry. Lectins shut off the production of digestive chemistry to allow a viable seed to survive travel through the digestive tract. I choose to calm and quiet the Immune System. Why would I want to stimulate an out-of-control immune response.

No! Stimulating your immune system will not reset it back to normal when it is out-of-control, responding inappropriately.

Do you need help? Are you sick and tired of feeling sick and tired? Are you frustrated with your boiled chicken meals. Oh, by the way, that bone broth you are drinking may be causing part of your problems as it stimulate cytokine that do not need antibody to flair up.

What are the Symptoms of Insulin Resistance?

Still Under construction

Insulin Resistance causes many symptoms easily diagnosed as many other conditions.

Symptoms of Insulin Resistance

Fatigue. Sometimes the fatigue is physical, but often it is mental. 

  • Muscles, nerves, brain, and all other parts of the body require oxygen and glucose to function efficiently. Insulin Resistance deprives all of them of the necessary oxygen and glucose. 
  • RBC agglutination slows blood flow to the muscles, nerves and all parts of the body. RBC agglutination causes RBCs to shed oxygen. Further depriving all of the body of oxygen and glucose. 

Brain fogginess and inability to focus. 

  • Brain requires oxygen and glucose to function efficiently. Insulin Resistance deprives the brain of the necessary oxygen and glucose. 
  • RBC agglutination slows blood flow to the muscles, nerves and all parts of the body.
  • RBC agglutination causes RBCs to shed oxygen. Further depriving all of the body of oxygen and glucose. 

Depression

  • When the brain is deprived of oxygen and glucose, brain function slows down, going into power saver mode creating the symptoms of “depression”. 
  • Brain requires oxygen and glucose to function efficiently. Insulin Resistance deprives the brain of the necessary oxygen and glucose.  
  • RBC agglutination slows blood flow to the brain. Further depriving the brain of oxygen and glucose. 

Nerve Pain – Blood/Nerve Reciprocity

Nerve pain occurs when nerves begin deteriorating from a lack of oxygen and glucose. This starts with Red Blood Cells clumping together and shedding oxygen caused by Insulin Resistance. This results in a failure to deliver oxygen and glucose to the nerves of the arms, hands, legs and feet.
  • Nerves and all other parts of the body require oxygen and glucose to function efficiently. Insulin Resistance deprives all of them of the necessary oxygen and glucose. 
  • All nerves depend wholly upon the arterial system for their oxygen and glucose and the quality of their function, such as sensation, signal transmission and motion.
  • A loss of blood flow results in nerve cell deterioration and ultimately in nerve death starting within minutes. A deteriorating nerve send pain signals to the brain.
  • RBC agglutination slows blood flow to the nerves and all parts of the body.
A failure to deliver oxygen and glucose to the nerves of the arms, hands, legs and feet causes them to start degenerating within minutes. This degeneration attracts Immune Cells to clear the damaged parts of the nerves. This is felt as pain occurring in the Ischemic area of the body.

High Blood Sugar

  • Mild, brief periods of low blood sugar are normal during the day, especially if meals are not eaten on a regular schedule. But prolonged hyperglycemia with some of the symptoms listed here, especially physical and mental fatigue, are not normal. 
  • Feeling agitated, jittery, moody, nauseated, or having a headache is common in Insulin Resistance, without immediate relief once food is eaten.

Intestinal bloating.

  • Most intestinal gas is produced from food fermenting and putrefying due to inadequate production of digestive chemistry.
  • Insulin Resistance sufferers who eat carbohydrates suffer from gas, lots of it. 

Sleepiness

  • Many people with Insulin Resistance get sleepy immediately after eating a meal exceeding their Carbohydrate Tolerance. Body is going into power-saver mode like your electronic device.
  • Fatigue after meals, craving sugar after meals, must have dessert: 
    • Cut back on carbs until you are no longer sleepy after meals and/or do not crave sugar after eating.

What is Insulin Resistance?

Insulin resistance is when cells in your muscles, fat, and liver don’t respond in a good way to insulin and can’t use glucose from your blood for energy. To make up for it, your pancreas attempts to make more insulin. Over time, your blood sugar levels go up, unless the high circulating insulin levels are further challenged by a prolonged period of fasting or dietary restriction, i.e. food allergies/sensitivities, Keto, Paleo, Vegan, etc. 

Why is your body not responding to insulin?

It is always assumed that the pancreas is producing an adequate quantity and quality of insulin. This may be accurate initially. However, as alkaline-activated pancreatic enzymes are activated within the pancreas, these enzymes begin digesting the insulin and glucagon producing endocrine cells. These enzyme them have access into the blood stream where they being breaking down the connective elastic lining of the blood vessels.

insulin resistance many common symptoms

You could be insulin resistant for years without knowing it. This condition causes ubiquitous symptoms that are common to more popular conditions, i.e. Low Thyroid, PCOS, Low T, High Cholesterol, etc. You can’t tell that you have insulin resistance by how you feel. The symptoms are common to all the popular hormone and cholesterol conditions.

Insulin resistance is often present for up to 10 years or more before an “official” diagnosis of Diabetes. Many never develop Type 2 Diabetes but suffer for decades from insulin resistance. Especially, those with long-term dietary restrictions. That is a long time to do nothing about it. It is also a long time for you to do something about it.

Waiting results in more prescriptions per person.

It is money in the bank for Doctors, Hospitals and Drug Companies to overlook Insulin Resistance, AKA: Pre-Diabetes. While you wait, they can prescribe Hormone Replacement Therapy, Thyroid Hormone Replacement, Cholesterol Lowering Drugs and Pain Relieving Drugs before they get to the real cash cow – Diabetes. Most will never develop Diabetes, but they will have you on these drugs for the remainder of your life unless you take action. Insulin Resistance and all of the ubiquitous symptoms are easily reversed. But they will never tell you that. Follow the money.

But My Blood Sugar Is Low Normal!

How many people are restricting their diet because of Professional and Social Media Influencer diet recommendations?

People with pre-diabetes or insulin resistance also can have low or normal blood sugars, if their high circulating insulin levels are further challenged by a prolonged period of fasting or dietary restriction, e.g. food allergy/sensitivities, Paleo, Keto, Vegan, Vegetarian, etc.