Dr Syihabuddin (Din Kapak Abdullah), seorang pegawai perubatan, sekali lagi mencabar saya untuk menjawab persoalan, dan kali ini berkenaan kajian in vitro oleh Kumar et al. Saya sudah banyak menjawab persoalan beliau namun beliau tidak menjawab soalan-soalan saya. Saya pun hairan siapa doktor, dia doktor ke saya doktor?
Samada Syihabuddin rasa malu atau tidak setelah terbukti beliau membohong berkenaan kajian in vitro Caly et al, yang pasti dia amat marah dengan saya dan dengan sesiapa sahaja yang menegur beliau atas pembohongan, percakapan putar belit dan perangai beliau.
Beliau terjumpa sebuah artikel kajian in vitro yang mendakwa ivermectin kononnya tidak berkesan melawan covid. Syihabuddin posted the following screenshot of the article in the comments section of my Facebook page:
Syihabuddin is thrilled that after much googling, he managed to find an article that claims that ivermectin was not effective in an in vitro study.
Who funded the study?
The funders of the study were the University Medical Center of Groningen (UMCG) and Marie Skłodowska-Curie. UMCG is involved in a European project on accelerating vaccine development. The project, Inno4vac, is a public-private partnership, jointly financed by the EU and vaccine producers, which is being coordinated by the European Vaccine Initiative in Germany. Marie Skłodowska-Curie also funded a review on mRNA vaccine development, the paper of which was published in February 2021 titled An Update on Self-Amplifying mRNA Vaccine Development. So it would be no surprise if the study was designed to fail.
First let’s see have a look at the status of the paper:
As we can see, the article is a preprint that has not been peer-reviewed. What does that mean? It means as follows:
As stated above, the article has not been finalized by authors, might contain errors, and report information that has not yet been accepted or endorsed in any way by the scientific or medical community.
It’s been a few months since the paper was published and obviously in vivo studies had proceeded and been completed long before it was published. In other words, this article is pretty much obsolete, since we have been overtaken by events, and was just another poor attempt at trying to denigrate ivermectin after Caly’s in vitro study, which proved that ivermectin inhibited the virus from docking with the cell and from replicating. In fact, many studies still went ahead after the article was published, which means the article did not cause any excitement except among the ivermectin deniers, such as Syihabuddin.
Anyway, what did other scientists have to say about the article?
Now, the title of the paper as well as the caption of section 3.3 shared by Syihabuddin appear misleading. The title of the article reads “Moxidectin and ivermectin inhibit SARS-CoV-2 replication in Vero E6 cells but not in human primary airway epithelium cells” while the caption of section 3.3 reads “Moxidectin and ivermectin do not exhibit anti-SARS-CoV-2 activity in Calu-3 and primary human bronchial epithelial cells”. These are statements in the absolute, which then contradicts what is also said in the article:
“To our surprise, however, no significant reduction in viral titers was observed following infection in presence of moxidectin and ivermectin, indicating that at these experimental conditions, the compounds did not exhibit an antiviral effect in Calu-3 cells.”
“Moxidectin and ivermectin have no significant effect on SARS-CoV-2 infection in human-derived cell models”
In the context of trying to influence readers, there is a big difference between not inhibiting or not exhibiting an effect versus not signficantly inhibiting or exhibiting an effect. There is a big difference between no effect and no signficant effect. Based on this in vitro study it is a fact, according to the authors themselves, that ivermectin did inhibit and did have an effect except that they do did consider it to be statistically significant — and as mentioned above (Ivermectin study #81), the values were not stated.
As stated above, Calu-3 resembles serous gland cells. To elaborate further:
Calu-3 is a non-small-cell LUNG CANCER cell line that grows in adherent culture and displays epithelial morphology. (This cell line was established in 1975 from a metastatic site (pleural effusion) in a 25-year-old Caucasian male with adenocarcinoma of the lung).[MSKCC]
Of 12 cell lines derived from human LUNG CANCERS, only Calu-3 cells showed high transepithelial resistance (Rte) and increases in short-circuit current (Isc) in response to mediators. [Shen et al]
Serous cells are specialized to secrete an enzyme solution. Examples include serous cells of the salivary glands, exocrine cells of the pancreas, gastric chief cells, and Paneth cells of intestinal crypts. Serous cells of the pancreas and the salivary glands are typically organized into secretory units called acini. [SIU]
Basically, the results of using Calu-3 in vitro cannot be used to determine the effective dosage of ivermecton in humans. I had explained to Syihabuddin in mid-September that Dr Pierre Kory had said:
“When they repeated the experiment in lung alveolar cells, they have achieved inhibitory concentrations in alveolar lung cell models easily. Article is under peer review.”
Obviously Syihabuddin fails to ackowledge that there is a big difference between human lung alveolar cells and Calu-3 in this respect.
Alveolar macrophages are the most abundant innate immune cells in the distal lung parenchyma, located on the luminal surface of the alveolar space. They are the first to encounter incoming pathogens and pollutants and to help orchestrate the initiation and resolution of the immune response in the lung. Similar to other tissue-resident macrophages, alveolar macrophages also perform non-immune, tissue-specific, homeostatic functions, most notably clearance of surfactant. [Joshi et al]
Human alveolar macrophages (HAM) are cells that may play a major role in host defense, and conversely display significant phlogistic potential. [Levy et al]
To maintain a proper balance, the lung needs specialized cells that can efficiently initiate and resolve inflammatory responses. Alveolar macrophages (AMs) and lung epithelial cells (ECs) are described in the literature as being the most important cells in the maintenance of lung homeostasis. AMs are the main immune cell type in the lung that determine the orientation and the magnitude of the immune response. In addition, they eliminate pathogens, apoptotic cells, and debris. [Bissonnette et al]
For example, unlike Calu-3, human alveolar macrophages have 15-Lipoxygenase.
12/15-lipoxygenase (12/15-LOX) is an enzyme, which oxidizes polyunsaturated fatty acids, particularly omega-6 and -3 fatty acids, to generate a number of bioactive lipid metabolites. A large number of studies have revealed the importance of 12/15-LOX role in oxidative and inflammatory responses. The in vitro studies have demonstrated the ability of 12/15-LOX metabolites in the expression of various genes and production of cytokine related to inflammation and resolution of inflammation. [Singh & Rao]
The above citations are for us to appreciate the significance of the difference of using Calu-3 vs lung alveolar cells.
In summary, the results of this in vitro study on Vero-E6 cells (monkey kidney cells) is consistent with the results of Caly’s study i.e. ivermectin inhibits the covid-19 virus from docking with the cell and preventing replication. Meaning, Caly’s study was not a fraud nor misleading. However, Kumar concludes that the effect on Calu-3 is not statistically significant. This is not surprising for the reasons explained above while the reality is different for when ivermectin is taken orally by humans.
All of the above notwithstanding, as has been explained in my previous post:
“The in vitro potency of ivermectin against Covid-19 virus is a testimony that this drug can be utilized to manage those patients who have been infected with SARS-CoV-2. Since the conditions in which the virus replicates and infects the cells in vivo and in vitro DIFFERS, a DECISIVE COMMENT about how ivermectin may prove to be beneficial to the patients CANNOT BE CONSTRUCTED YET.” — Dhyuti Gupta, Ajaya Kumar Sahoo, Alok Singh, Department of Pharmacology, All India Institute of Medical Sciences,Raipur, Chhattisgarh, India
— Ivermectin: potential candidate for the treatment of Covid 19, Brazilian Journal of Infectious Diseases
CONCLUSION
Kumar’s in vitro study in actual fact proves that ivermectin is an inhibitor in both monkey kidney cells and Calu-3 except that Kumar’s considers the effect to be not statistically signficant in the case of Calu-3 — but exhibits antiviral activity nonetheless. Therefore, this study does nothing to nullify in vivo studies that have been done based on normal dosage. There may have been a little doubt had Kumar used lung alveolar cells instead of Calu-3 and obtained the same result. It is as if the study was designed to fail by using Calu-3 instead of lung alveolar cells. Even when Kumar said the following in the paper, we can detect that it does not sound sincere:
“Both moxidectin and ivermectin exhibited antiviral activity in Vero E6 cells. Subsequent experiments revealed that the compounds predominantly act on a step after virus cell entry. Surprisingly, however, in human airway-derived cell models, moxidectin and ivermectin failed to inhibit SARS-CoV-2 infection, even at a concentration of 10 μM. These disappointing results calls for a word of caution in the interpretation of anti-SARS-CoV-2 activity of drugs solely based on Vero cells. Altogether, these findings suggest that, even by using a high-dose regimen of ivermectin or switching to another drug in the same class are unlikely to be useful for treatment against SARS-CoV-2 in humans.” – Kumar et al
Do not think that scientists/doctors and universities are free from bias. Syihabuddin is an example of a government doctor who has been proven to be not only biased but also a liar. But even then, Kumar couldn’t hide the fact that ivermectin did show antiviral activity in Calu-3, even if concluding the results were not statistically signficant.
Nanti Syihabuddin akan cari lagi kajian in vitro atau in vivo yang lain untuk mencabar saya menjawab sekali lagi to divert attention from his shortcomings dan untuk mengelak dari menjawab soalan-soalan saya. In this respect, Syihabuddin likes to cherry-pick stones while ignoring the mountain.
– AA –
Azmi Arshad 