Archive for the ‘research’ Category

Anti-Borrelia Antibody Staining of Cultured Biofilms in Human Blood

https://www.linkedin.com/pulse/anti-borrelia-antibody-staining-cultured-biofilms-human-peter-kemp/

Fluorescent antibody stained agent
Fluorescent antibody stained agent

ANTI-BORRELIA ANTIBODY STAINING OF CULTURED BIOFILMS IN HUMAN BLOOD

Peter Kemp
Peter Kemp
Private researcher at Private borreliosis research

INTRODUCTION

Antibody staining can detect specific proteins when a sample contains matching antigens. This experiment used polyclonal antibodies to borrelia burgdorferi targeting at least 5 different proteins. The antibodies used are proprietary off-the-shelf products conjugated with the popular FITC fluorescent molecule and sold by KPL and Abcam. When FITC is excited with blue light it fluoresces and emits green light. With the correct filtering, it is possible to observe only subjects which are fluorescing at specific wavelengths including agents to which the antibodies have bound.

SAMPLE DONORS

All seven UK adult blood sample donors were chronically and long-term ill and six had previously been diagnosed with Myalgic Encephalomyelitis (M.E.). All had symptoms which correspond with Lyme disease.

METHOD

Donors provided 4mls of venous blood in lavender-top vacutainer tubes with EDTA. The blood collection tubes were kept upright for several hours to allow the cells to settle and achieve a degree of cell layering. Gentle centrifuging for 20 minutes at ~20 to 40g’s further compacted and layered the cells. Most of the plasma was drawn-off with a pipette without disturbing the cell layers. 1.5 to 2mls of BSK was gently added to the tubes, minimising disturbance of the cell layers. The tubes were kept at room temperature or incubated at 30°C. Incubation appears to speed the development of a biofilm. After 3 to 7 days all samples developed a bound cluster at the top of the cell layers. This was associated with a cohesive fluid more viscous than the culture medium and which appeared clear to the naked eye. Under the microscope it was slightly milky. The cluster and its associated slime were obtained by removing most of the BSK with a pipette. The cluster could then be ‘grasped’ with a 3ml disposable pipette and with the tube tilted to ~45° or more – was carefully dragged up the side of the tube and deposited on a slide. Sometimes it was easier to grasp the cluster with tweezers, but these have to be very narrow to reach into the tube. Each sample was usually sufficient to prepare 3 – 4 slides. To each slide a drop of FITC conjugated anti-borrelia burgdorferi conjugated antibody, pre-diluted at 10 or 20 to 1, was added and gently mixed.

RESULTS

The results are presented as pairs of micrographs. The top photo in each matched pair, show a normal darkfield image with only the fluorescence emission filter in place. The bottom photo of the pair shows the same field of view with the fluorescence excitation filter installed. In some images refocusing may have slightly offset some subjects. The images can be seen here:  https://www.flickr.com/photos/76898309@N08/albums/72157713385691712

DISCUSSION

The experiment requires replication which includes healthy controls which I will not conduct.

It is notable that in experiments with freshly drawn or stored whole blood, minimal signs of staining with anti-B.b antibodies occurs. After simple culture with BSK, some white blood cells appear to contain granules that react and aside from these a few tiny agents are seen to have stained. It is only after culture with BSK using the method described above that spectacular staining occurred. The culture method layers white blood cells and probably a good number of platelets at the top of the sample. This is where the slime substance is found and where the clumped blood cells contain and/or are surrounded by antigenic material. This suggests the possibility that some white blood cells may be reactive. However, further experiments showed that with lymphocytes, eosinophils and basophils, only a small proportion contained granules within the cytoplasm that bound the antibodies.

Replication of the experiment with monoclonal antibodies could help to determine whether a cross-reaction is occurring, and potentially improve efficiency by identifying which particular B.b. proteins are being expressed, if any.

All donors in the current experiment had been ill and symptomatic for many years, mostly for over a decade. If replication as suggested proved the method for identifying infection with B.b., further experiments with shorter term infections would be required to identify within what time range a 4ml blood sample would be reactive.

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For more:  https://madisonarealymesupportgroup.com/2018/09/27/spirochete-culture-microscopy-videos-see-whats-inside-you/

https://madisonarealymesupportgroup.com/2020/03/02/spirochetes-in-sputum-of-chronic-lyme-patient/

https://madisonarealymesupportgroup.com/2018/09/29/microscopy-of-spirochaete-biofilm/

https://madisonarealymesupportgroup.com/2019/02/27/advanced-imaging-found-bartonella-around-pic-line/

Category:

Lyme, research, Testing

How the COVID-19 Pandemic Will End

https://jameslyonsweiler.com/2020/03/15/how-the-covid-19-pandemic-will-end/

James Lyons-Weiler, PhD – 3/15/2020

How this pandemic will end depends on how soon the medical community realizes that therapeutics are 100% essential this time.

NOW THAT SOCIAL DISTANCING is occurring in the US and other countries, and some countries are under general quarantine, what next? How long will social distancing and quarantine be in effect?

The promise of a vaccine against Coronavirus is too far off to play a role in ending the deadly and economy- smashing effects of our response to COVID-19. The move to start human trials of SAR-CoV-2 vaccines without proper animal safety studies has prompted outcries (See Moderna and US NIAID Poised to Endanger the World Population?) given the reality that SARS-CoV studies had unacceptable safety issues of pathgenic priming : animals vaccinated using Spike protein based vaccines against SARS and MERS – close cousin of SARS-CoV-2 – had worse outcomes when challenged with the virus that the vaccine was supposed to protect them against. Even the most vocal vaccine proponents such as Paul Offit and Peter Hotez now say skipping the animal studies could result in unacceptable risks.

I should point out that this week, they have joined the ranks of people using freedom of speech to “spread” vaccine skepticism.

Given the catastrophic outcomes in animal safety testing for the SARS coronavirus vaccine, we must understand that the current vaccine trials will likely fail. We must realize also that social distancing alone will be insufficient to bring a reasonably quick end to the need for social distancing and quarantine.

Here are some simulation result of the outcome of using either just social distancing to manage the spread of SARS-CoV-2 or combining social distancing with therapeutics:

Clearly therapeutics will be necessary to play a role in bringing about an end to this pandemic. Resources on this are listed at the end of this article.

Here I list feasible steps that are absolutely essential for the counties to end the pandemic as quickly as possible.

1. Testing. Develop local testing capacity. Avoid CDC’s flawed test until they produce data showing high sensitivity and specificity of their primer probe sets. Testing will be relevant towards effective resource use, but people should not wait until they have symptoms to take steps to reduce the severity of virus-related illnesses.

The utility of accurate testing should be a lesson for all respiratory virus-related illnesses and perhaps we will see a massive paradigm shift towards accurate testing and reporting of which respiratory virus individuals actually have and perhaps medicine will now abandon so-called “flu syndrome”.

2. Plasma convalescent therapy. Each test-positive patient who has recovered should be asked to donate a liter of blood from which antibodies can be harvested. The resulting product should be screened for other viruses and then administered to those on the front lines first – nurses, MDs, medical staff – EMTS, the military, all essential personel in the “infrastructure” industries and in the food industries.

3. Therapeutics (Prophylatic Treatments). Mass production and distribution of other therapeutics including antivirals that have shown to be effective against SARS-CoV-2 for prophylactic use in families and workplaces who have a members or co-workers who test positive for COVID-19. Herbal remedies that also show efficacy should not be discouraged – anything that might help but will not hurt should be used. A registry of treatment experiences and outcomes should be created to help identify the most efficacious treatment options.

4. True Immune Enhancement. Increase the use of supplements that enhance and strengthen the immune system. These include vitamins such as vitamin D vitamin A and large doses of vitamin C. Encourage the uptake of micronutrients including zinc and selenium which have specifically been shown to inhibit the entry of SARS and SARS-CoV-2 into human cells. An extract of licorice root appears to have some effect. (The phrase “immune enhancement” is being misused to refer to a dangerous outcome due to exposure to the virus following vaccination, which should be referred to as “Pathogenic Priming”.

5. Wellness. Increase life a affirming and healthy lifestyle choices including exercise, exposure to sunlight, daily air exchange of the home and healthy avocational activities. People should be encouraged to take the time for early spring cleaning, take up a new exercise regime such as thai chi, to learn a new language or to learn a new musical instrument.

Use this time to exercise to reduce metabolic syndrome and diabetes is essential to reduce your risk of mortality. Diabetics have a 6% fatality risk. People with cardiovascular illness including hypertension have a 10% case fatality risk. Talk to your doctor about moving from ACE inhibitor blood pressure management to other options. Also talk to your doctor about adding exercise to your lifestyle to help control your diabetes.

I applaud the move taken by the banking and and finance industries to help stabilize hardships visited upon families by loss of work including interest forgiveness on loans, tolerance of delayed, late or skipped payments. Mental wellness is essential for making optimal life choices.

It is time to re-discovery the best part of ourselves. Engage in charitable acts to aid the elderly and those at most risk of serious illness.

Resources on Therapeutics

Antivirals

Treatments under study include nutriceuticals and supplements; ” and antiviral drugs such as Disulfiram and Chloroquine Phosphate.

Potential Inhibitor of COVID-19 Main Protease from Several Medicinal Plant Compounds by Molecular Docking Study. https://www.preprints.org/manuscript/202003.0226/v1/download

See: “Disulfiram can inhibit MERS and SARS coronavirus papain-like proteases via different modes“.

See “Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro“.

See “Expert consensus on chloroquine phosphate for the treatment of novel coronavirus pneumonia“.

The race to find a coronavirus treatment: One strategy might be just weeks away, scientists say https://www.usatoday.com/story/news/health/2020/03/15/coronavirus-treatment-could-coming-doctors-say-still-no-vaccine/5052788002/

Chloroquine Confirmed Effective as Coronavirus Cure https://www.hngn.com/articles/228138/20200223/coronavirus-cure-chloroquine-confirmed-effective.htm

Vitamins and Minerals

See: How doctors say you can boost your immune system to protect against flu, coronavirus https://www.abcactionnews.com/news/national/coronavirus/how-doctors-say-you-can-boost-your-immune-system-to-protect-against-flu-coronavirus

Herbals and Supplements

Selenium and other micronutrient deficiency is considered to play a role in severity of a coronavirus infection (See: “Micronutrient Selenium Deficiency Influences Evolution of Some Viral Infectious Diseases“). Glycyrrhizin, an active component of liquorice roots, has been found to have few toxic effects and to be clinically effective against SARS-associated coronavirus. (See: See “Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus“)

Supplements that might have a positive effect include N- acetyl cysteine, selenium, spirulina and high dose glucosamine (See “Nutraceuticals have potential for boosting the type 1 interferon response to RNA viruses including influenza and coronavirus

Colloidal Silver Nebulizer

See drpaulapproved.com for info on colloidal silver nebulizer.

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**Comment**

Please know that antivirals are not without risk.

For more:  https://madisonarealymesupportgroup.com/2020/03/15/corona-virus-common-sense/

https://madisonarealymesupportgroup.com/2020/03/15/herbal-treatment-for-coronavirus-infections-buhner/

https://madisonarealymesupportgroup.com/2020/03/10/corona-virus-lyme-disease-drs-bock-rawls/

https://madisonarealymesupportgroup.com/2016/03/28/combating-viruses/

 

 

Category:

research, Supplements, Treatment, vaccines, Viruses

Vector Competence Studies With Hard Ticks & Borrelia Sensu Lato Spirochetes: A Review

https://www.ncbi.nlm.nih.gov/pubmed/32067949

2019 Dec 14:101359. doi: 10.1016/j.ttbdis.2019.101359. [Epub ahead of print]

Vector competence studies with hard ticks and Borrelia burgdorferi sensu lato spirochetes: A review.

Eisen L1.

Abstract

Use of emerging technology allowing for identification of genetic material from pathogens and endosymbionts in ticks collected from humans, domestic animals, wildlife, or the environment has resulted in an avalanche of new data on tick-microorganism associations. This rapidly growing stream of new information is a tremendous resource but also presents challenges, including how detection of pathogen genetic material in ticks should best be interpreted. There is a tendency in the more recent published literature to incorrectly use the term “vector” based on detection of pathogen genetic material from tick species not experimentally confirmed to serve as vectors of the pathogen in question.

To serve as a vector of a horizontally maintained pathogen, such as a Borrelia burgdorferi sensu lato (s.l.) Lyme borreliosis spirochete, the tick species in question must be capable of acquiring the pathogen while feeding in the larval or nymphal stage on an infectious host, maintaining it transstadially through the molt, and then transmitting the pathogen to a naïve host while feeding in the subsequent nymphal or adult stage.

This review examines the experimental evidence for and against species of hard (ixodid) ticks from different genera to serve as vectors of B. burgdorferi s.l. spirochetes.

  • Of the 18 Ixodes species ticks evaluated to date, 13 were experimentally confirmed as vectors of B. burgdorferi s.l. spirochetes.
  • These studies focused primarily on the three major Lyme borreliosis agents: Borrelia burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii.
  • In striking contrast, none of 8 tick species from other genera (1 Amblyomma species, 5 Dermacentor species, and 2 Haemaphysalis species) evaluated to date were unequivocally experimentally confirmed as vectors of B. burgdorferi s.l. spirochetes.

The strength of the evidence for or against each tick species to serve as a vector of B. burgdorferi s.l. spirochetes is discussed together with key knowledge gaps and research challenges.

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**Comment**

This author, a CDC employee, is basing his findings on previous research.  There’s nothing new here.  

Also of import is the fact borrelia has been found in other ticks – just not enough for to be “statistically” important, OR in the instance of the Lone Star tick (Amblyomma americanum) there has been hot debate as to whether STARI is Lyme or not.  Patients and advocates alike claim the symptoms are one in the same.  The debate continues.

The trouble is, what if you are the poor sucker that gets bitten by that “statistically” insignificant tick?  Well, you lose because mainstream doctors are going to skim this review and conclude that ONLY certain ticks transmit Lyme/borrelia. They are going to write you off as psychosomatic:

Important excerpt:

Yes, vector competence was confirmed experimentally; No, vector competence was evaluated experimentally but could not be confirmed; Blank space, tick species not yet evaluated for this B. burgdorferi s.l. species.

There were numerous places where competence couldn’t be confirmed (which is a far cry different from it can’t happen) as well as the fact there were tons of blank spaces – which means the tick species hasn’t even been evaluated yet.

Here’s the dealeo, all ticks transfer fluid.  ALL TICKS should be suspect.  Period.  Remember all the research I’ve posted stating Lyme “didn’t exist” in various geographical places until someone pushed hard enough to get it recognized:  https://madisonarealymesupportgroup.com/2018/05/31/no-lyme-in-the-south-guess-again/

https://madisonarealymesupportgroup.com/2016/09/24/arkansas-kids-denied-lyme-treatment/

Excerpt:

According to Dr. Naveen Patil, Director of the Infectious Disease Program, ADH,

“We don’t have Lyme Disease in Arkansas, we have the ticks that transmit Lyme Disease but we don’t have any recorded cases of Lyme Disease.” 

We can thank a mother from Arkansas for getting Arkansas on the map for Lyme disease.

And so it goes:  https://madisonarealymesupportgroup.com/2017/10/24/no-lyme-in-oklahoma-yeah-right/

https://madisonarealymesupportgroup.com/2019/04/22/its-just-crazy-why-is-lyme-disease-treatment-so-difficult-to-find-in-mississippi/

https://madisonarealymesupportgroup.com/2019/11/25/hundreds-of-people-impacted-by-tick-borne-illnesses-in-north-carolina/

Tick research is similar in that until something gets documented (published), researchers and doctors alike treat it as if it’s never, ever happened, and therefore (circular reasoning) it won’t happen in the future.

Category:

research, Testing, Ticks, Transmission

Unusual Case of Suspected Lyme Disease in Patient With Ear Pain

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6906891/

. 2019 Oct-Dec; 11(4): 160–162.
Published online 2019 Nov 26. doi: 10.4103/jgid.jgid_75_19
PMCID: PMC6906891
PMID: 31849437

An Unusual Case of Suspected Lyme Neuroborreliosis in a Patient Presenting with Ear Pain

Michael Anthony Ramada, Saman Kannangara,1 and Hany Eskarous1

Abstract

We report a case of suspected Lyme neuroborreliosis (LNB) where the patient’s sole symptom was chronic, intermittent episodes of unilateral ear pain. This case was unusual because this patient did not show any other neurological or extraneurological symptoms of Lyme disease. LNB is a manifestation of infection by Borrelia burgdorferi spirochetes that can manifest in a number of different clinical presentations depending on the nervous system dissemination of the bacteria. Most commonly, these neurological findings present as peripheral nerve radiculopathies, but rarer involvement of the central nervous system (CNS) can occur if a patient goes untreated. Most often, CNS involvement presents as meningitis with increased lymphocytes but can, in rarer cases, involve the spinal cord or brain parenchyma.

The diagnosis of LNB was made after the patient was found to have suspicious lesions in the spinal cord and brain parenchyma on magnetic resonance imaging as well as cerebrospinal fluid Borrelia antibody index was consistent with CNS Lyme disease. We discuss this case as a unique clinical presentation of suspected LNB and the diagnostic findings associated with this infection.

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**Comment**

Nothing surprises me about Lyme disease anymore.  “Not everything is Lyme, but Lyme CAN be anything.”  Truer words were never spoken.

 

 

Category:

Lyme, research, Testing

Dogs With Proteinuria Found to Have Exposure to Rickettsia, Ehrlichia, and Lyme Disease

https://www.ncbi.nlm.nih.gov/pubmed/31916316/

2020 Jan 8. doi: 10.1111/jvim.15610. [Epub ahead of print]

A retrospective study of vector-borne disease prevalence in dogs with proteinuria: Southeastern United States.

Purswell EK1, Lashnits EW1, Breitschwerdt EB1, Vaden SL1.

Abstract

BACKGROUND:

Proteinuria is a risk factor for progressive kidney injury in dogs. Enhanced understanding of potential associations between canine vector-borne diseases (CVBD) and proteinuria is needed.

OBJECTIVES:

To determine the proportion of evaluated proteinuric dogs exposed to ≥1 CVBD, including Babesia spp., Ehrlichia spp., spotted-fever group Rickettsia, Bartonella spp., Anaplasma spp., hemotropic Mycoplasma spp., Borrelia burgdorferi, and Dirofilaria immitis, and to determine if demographic or clinicopathologic differences exist between proteinuric dogs exposed to CVBD versus proteinuric dogs with no evidence of CVBD exposure.

ANIMALS:

Two-hundred nine proteinuric dogs, concurrently tested for CVBD, which were examined at a single academic veterinary hospital between January 2008 and December 2015.

METHODS:

Retrospective cross-sectional study. Demographic, clinicopathologic, and CVBD test results were extracted from medical records. A multivariable logistic regression model was used to assess associations between CVBD and selected variables.

RESULTS:

Based on serology and polymerase chain reaction testing,

  • 34% of proteinuric dogs (72/209) were exposed to ≥1 CVBD
  • Exposure to Rickettsia spp. (19%)
  • Ehrlichia spp. (12%)
  • B. burgdorferi (9%)

The CVBD exposure was lower in dogs tested in autumn or spring, higher in intact dogs, and higher in dogs with lower serum albumin and higher serum creatinine concentrations.

CONCLUSIONS AND CLINICAL IMPORTANCE:

Exposure to CVBD, particularly exposure to Rickettsia spp., Ehrlichia spp., and B. burgdorferi was found in proteinuric dogs from the southeast United States. Additional controlled prospective studies examining a potential causal relationship between CVBD and proteinuria are warranted.

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**Comment**

The question of course is – does this also happen in humans and to what degree?

 

Category:

Activism, Ehrlichiosis, Lyme, research, Rickettsia