It’s the academia, dummy! or when quantity supersedes quality

Many of us, faculty members dedicated to the profession of university professor, began our professional careers investing the best years of our youth in academically preparing ourselves in the best way possible. We continued our careers by searching for a renowned institution to work for. Its prestige provided us with a solid foundation for quick growth, thus reaching the top of the academic hierarchy, making us feel that we made the best investment for our future. Since it is our responsibility to prepare ourselves, it is also our responsibility to make sure this preparation bears the best fruit for society and our families. But also, we aspire to heighten our prestige, even a little, so that at the dusk of our professional life we can feel the satisfaction of having accomplished our mission in society and be rewarded with the appreciation and respect of our colleagues (which results in sincere affection), who we also respect and admire, and fulfill the debt to our family who supported us and sacrificed so much for our eagerness to pursue professional success, a success that has only one noble goal: academic excellence

Origin of personalized medicine in pioneering, passionate, genomic research

Personalized medicine, one of the main promises of the Human Genome Project (HGP) that began three decades ago, is now a new therapeutic paradigm. With its arrival the era of developing drugs to suit all patients, yet often having to withdraw a promising new one because a minority of patients was at risk, even though it had proved valuable for the majority was consigned to history as were trial-and-error strategies being the predominant means of tailoring therapy. But how did it originate and the earliest examples emerge? Is it true that the first personalized diagnostic test was the companion test for Herceptin®? This account of a remarkable journey from genomic and translational research to therapeutic and diagnostic innovations, describes how sequencing the human growth hormone (hGH) locus provided proof of principle for HGP-inspired personalized medicine. Sequencing this locus and the resultant biomanufacture of HGH and the development of a test capable of detecting which patients would benefit from its administration helped silence the skeptics who questioned the validity of such an approach. The associated companion diagnostic was created four years before the invention of the HercepTest® (registered as the first companion diagnostics ever developed). By cultivating genomic research with passion and pursuing its applications, we and many others contributed to the emergence of a new diagnostics industry, the discovery of better actionable gene-targets and to a revitalized pharmaceutical industry capable of developing safer and more effective therapies. In combination, these developments are beginning to fulfill the promise of the HGP, offering each patient the opportunity to adopt the right treatment at the correct dosage in an opportune manner

The Role of the Human Growth Hormone Gene Family in Pregnancy

A pregnant woman´s body undergoes profound anatomical and physiological changes to accommodate the needs of the maternal-fetal unit required for a successful pregnancy. During normal pregnancy, the placenta produces a variant of human growth hormone as well as a chorionic somatomammotropin hormone. These are the placental members of the human growth hormone gene family and play a crucial role in the regulation of maternal and fetal metabolism, as well as in the growth and development of the fetus. For this reason, the scope of this chapter is to describe the differences of the biochemical and physiological roles of the hormones coded in this locus during pregnancy, the repercussions of their deficiencies, and role in some of the most prevalent pathologies during pregnancy affecting either the mother or the fetus and also to describe how pioneering sequencing of this locus allowed our laboratory to invent the first companion diagnostics test and thus contributed to the dawn of the personalized medicine era

La reacción en cadena de la polimerasa a dos décadas de su invención

La PCR, tÈcnica inventada por Kary B. Mullis en 1983, emplea un par de oligonucleÛtidos para de- limitar una regiÛn de interÈs y una polimerasa para extenderlos, utilizando ambas hebras del gen en cuestiÛn como plantilla. Al repetir este ciclo dece- nas de veces, se duplica cada vez el fragmento de ADN en cuestiÛn. AsÌ se logra copiar millones de veces la secuencia de interÈs, aunque se encuentre entre millones de otras secuencias de ADN. A 20 aÒos de su invenciÛn, la PCR se cataloga como la estrella de las herramientas de la biologÌa mole- cular. En la actualidad son innumerables las aplica- ciones de su utilizaciÛn en m ̇ltiples campos de las ciencias biolÛgicas, agropecuarias y de la salud

Comparison of five commercial kits for SARS CoV 2 RT-PCR diagnosis.

Background: SARS-CoV-2 was identified as the causal agent of the COVID-19 pandemic. Its rapid spread and huge health and economic impact prompted the development of diagnostic tests to opportunely identify affected individuals as a prerequisite to quarantine them and avoid further spreading the infection. Methods: The following commercial RT-PCR kits, approved by our National Institute of Diagnostics and Epidemiological Reference of InDRE (from the Spanish name) were tested: Vircell (Granda, Spain), Light Mix Roche (Berlin, Germany), Logix Smart (Utah, USA), 1 Copy (Korea Republic), and RIDA GENE (Darmstadt, Germany). RNA was isolated either manually or automatically (QIAmp Viral RNA and SMART-32, DAAN GENE kits, respectively) from naso and oro pharyngeal swabs from suspicious individuals living in Mexico city and outskirts. Results: Since May 2020, when we received InDRE´s SARS-COV2 diagnostic approval, we have processed nearly 20,000 naso and oro-pharyngel swabs samples. The qualification of kits, as per their analytical performance, value of their controls, and convenience (mono versus multiplex) resulted in the following ranking from the most to the least convenient: 1) RIDA, 2) Vircell, 3) Roche, 4) 1 Copy, and 5) Logix Smart. Conclusions: Both, analytical performance and convenience to process quantious parallel samples in a short period of time, and particularly sensitivity, were key parameters for our laboratory to adopt either RIDA or Vircell kits. They are particularly useful in cases with low viral load, which even if asymptomatics, can be contagious for vulnerable subjects within their families, community, and at work

Production of codon optimized Polyomavirus small t antigen in Escherichia coli.

Background: Hauzen et al., went against the dogma that cancer only develops by genetic factors and postulated oncogenic human papillomavirus (HPV) as the etiological agent of cervical cancer (2008). The eleven new human polyomaviruses (HPyVs) identified have been shown to infect humans subclinically from an early age. The small t-antigen (tAg) proteins of polyomaviruses control cellular phosphorylation mechanisms leading to uncontrolled replication cycles and cancer. Mechanisms of transformation and progression to tumor cells, and cellular tropism of the new HPyVs, remain unknown. Availability of recombinant tAg (rtAg) can contribute to understanding them. Methods: tAg coding sequences derived from Merkel cell polyomavirus (MCPyV) genome were codon-optimized, synthesized, cloned in an expression vector (pGEX), and transformed into E. coli. Clones were fermented, induced for expression of the tAG cassette, and purified by IMAC (Immobilized metal affinity chromatography). Results: rtAg produced in bacteria from different expression strains demonstrated distinct expression levels. The rtAg of ~20KDa was produced at the level of 11.3 mg L-1 and folded correctly since antibody 5 anti-MCPyV recognized rtAg. Conclusions: MCPyV tAg was efficiently expressed in E. coli. The availability of rtAg from MCPyV will be useful in immunity diagnostics, structure studies, and investigation of metabolic pathways interference and cell tropism features of HPyVs infections. This especially until cell culture systems for new HPyVs are developed. Acknowledgements: Our sincere gratitude to the Dana-Farber Cancer Institute for gene and antibody samples as well as the National Council of Science and Technology (CONACYT) for the scholarship 304814 to LMRM

Chromium picolinate, biotin, and sodium bicarbonate combination as a dietary supplement in the treatment of type 2 diabetes

Background: Type 2 diabetes mellitus (T2DM) is characterized by hyperglycemia due to insulin resistance, which can lead to micro and macrovascular complications. The importance of glycemic control for prevention demands the need to promote accessible and safe treatments among others in the form of scientifically proven nutritional supplements. Previous studies have suggested that the consumption of bicarbonate-rich mineral water altered blood metabolites and gut microbiome which has beneficial effects on patients with T2DM. Likewise, chromium picolinate and biotin have shown usefulness in glycemic control. The objective of our study was to evaluate the supplementation with chromium picolinate, biotin, and sodium bicarbonate in patients with T2DM. Methods: We planned and supervised the execution of a crossover, randomized, double-blind, placebo-controlled study of patients with the diagnosis of T2DM that was conducted in Diabetes Clinics of the Endocrinology Service of the University Hospital “Dr. José E. González” of the Autonomous University of Nuevo Leon in Monterrey, Mexico from June 2011 to July 2012. Patients’ follow-ups during the study included a day-0 baseline visit and six more visits over the next six months. Efficacy of treatment was assessed by expressing changes in hemoglobin A1c (HbA1c), body mass index (BMI), and blood pressure (BP). Results: Forty-seven (62.6%) of the original 75 patients completed the trial. Regarding the baseline characteristics, 25 (53.1%) of the participants were male and the mean age was 55.23 ± 9.88. The mean HbA1c was 8.38 ± 1.08%, the mean BMI was 29.34 ± 4.64, the mean systolic BP of 143.84 ± 23.6 mm Hg, and the mean diastolic BP of 84.5 ± 12.13 mm Hg. When comparing the changes that occurred after both interventions, we observed that the HbA1c in the active ingredient group decreased (-0.15%) and in the placebo increased (+0.12%) (p=0.148). When we subdivided both groups according to their HbA1c level before the intervention and compared the participants with HbA1c ≥9, the placebo group had an increase of 0.15 ± 1.32 % and the reduction in the active ingredients was -0.68 ± 1.58 % (p=0.158). Conclusions: In our study, we observed that the supplementation with chromium picolinate, biotin, and sodium bicarbonate decreased HbA1c in 3 months compared to the placebo group in which there was an increase, but with a tendency in the statistical analyses. We believe that this could be due to two reasons: the size of our sample, due to the large percentage of participants who dropped out of the study, or because the treatment period to observe a greater difference should have been longer

Biobanking in NE Mexico for biomedical research and clinical needs

Background: The advancement of biomedicine demands tools that translate its achievements into services to both the scientific community and the pharma/biotech industry. Biobanks are powerful tools that collect, process, store, manage, and distribute biospecimens and their associated clinical/demographic data to users carrying out studies aimed at causing a real public health impact. For our laboratory to offer pharma/biotech companies support for their projects with the highest quality possible biospecimens we adopted the Best Biobanking Practices from ISBER (https://www.isber.org/). Methods: As a result of the pandemics, great effort has been dedicated to help the health ecosystem through a diagnostic service for SARS CoV-2 (by RT-PCR). Emphasis was put on proper sample collection, preanalytical characterization, and storage in ultra-low freezers. Their pre-analytical characterization included determining yield and purity by spectrophotometry using the NanoDropTM 2000 (Thermo-Fisher. Mexico City, Mexico). Results: We biobanked and supplied to internal (our Genetics laboratory) and external (validation protocols of pharma/biotech international companies) clients almost 2,000 RNA samples. Given the preanalytical qualification of the biospecimens, they performed satisfactorily for our clients’ diagnostic and innovation protocols needs. Conclusions: The biobanking services provided to both our diagnostic laboratory and to pharma/biotech companies that contracted our services delivered research materials of the highest quality. Being a private biobank recognized now nationally and internationally by public and private institutions has allowed us to participate in projects evaluating innovative diagnostics methods and devices

Correlation between Ct-values and symptoms of COVID-19 patients

Background: Currently available RT-PCR methods for the diagnosis of COVID-19 can give an estimate of the viral load. The cycle threshold value (Ct-value) of the PCR correlates inversely with the viral load; low Ct-values indicate high viral loads and vice versa. Higher viral loads have been seen to correlate with disease severity and infectivity. Therefore, we studied the correlation of the Ct-value of RT-PCR and the most common symptoms of COVID-19 individually. Methods: A prospective and descriptive study was carried out with the subjects that attended our laboratory for a COVID-19 test from September 14, 2020, to January 30, 2021. Subjects filled out a questionnaire with demographic and clinical information prior to taking the naso and oropharyngeal samples. The samples were processed by Vircell SARS-CoV-2 Real-time PCR Kit (Granada, Spain). Statistical analyses were performed using IBM SPSS software. Results: We included 657 positive subjects with complete information, with a median age of 36 (27-47) and a male predominance of 477 (72.6%). Of these, 395 (60.1%) were symptomatic and the median number of symptoms was 2 (0-5). The most predominant symptoms were headache 271 (68.6%), cough 229 (58%), and myalgias 180 (45.6%). The median Ct-value for gene N was 30 (23-36) and for gene E was 31 (23-35). In comparison between symptomatic and asymptomatic subjects, asymptomatic patients had a higher Ct-value (lower viral load) in both genes and a lower age (p Conclusions: The viral load correlates with symptoms within COVID-19, having found that higher viral loads were correlated with symptoms such as headache, cough, and fever, while lower viral loads were correlated with dyspnea, diarrhea, and alterations of smell or taste senses

Automated versus manual RNA isolation for the laboratory diagnosis of SARS-CoV-2.

Background: The rapid spread and huge health and economic impact witnessed even from the beginning of the COVID-19 pandemic prompted the development of diagnostic tests to opportunely identify individuals infected by its causing agent, the SARS CoV-2 virus. This is a prerequisite to quarantine them to avoid further spreading the infection specially to their vulnerable co-workers and family contacts. The golden standard for pathogen detection is the Polymerase Chain Reaction (PCR). To obtain an accurate result, it is important to carry out an optimal isolation of the viral RNA genome. Methods: This study aimed to compare manual (kits of Qiagen or DAAN) vs automatic (SMART-32 equipment of DAAN) RNA isolation methods. 372 samples were processed of which 200 were negative and 172 were positive of which 181 were processed manually and 191 automatically. Pre-analytical characterization of the RNA resulting from both methods included quantification of yield and qualification of purity by spectrophotometry in the Nanodrop (Thermo-Fisher, Mexico City). Results were comparatively evaluated employing the IBM SPSS Statistics software. Results: The median yield of RNA obtained by the manual method resulted higher than that rendered by the automatic method. Regarding purity (as judged by the ratios of A260/230 and A260/280) the manual method reflected better parameters than the automated one. On the other hand, when dealing with large amounts of samples, the latter was more convenient and faster. Conclusions: The manual method gives slighter better yield and purity than the automated one. However, quality wise, RNA from both methods is equally suitable for RT-PCR diagnosis of the SARS-CoV-2. The demand in the laboratory for processing large volumes in the minimal time, tips the scale to the automatic method