Phenotypic and Genotypic Drug Resistance of Mycobacterium tuberculosis Strains Isolated from HIV-Infected Patients from a Third-Level Public Hospital in Mexico.

BACKGROUND: Drug-resistant tuberculosis (TB) is associated with higher mortality rates in patients with human immunodeficiency virus (HIV). In Mexico, the number of deaths due to TB among the HIV-positive population has tripled in recent years. METHODS: Ninety-three Mycobacterium tuberculosis strains isolated from the same number of HIV-infected patients treated in a public hospital in Mexico City were studied to determine the drug resistance to first- and second-line anti-TB drugs and to identify the mutations associated with the resistance. RESULTS: Of the 93 patients, 82.7% were new TB cases, 86% were male, and 73% had extrapulmonary TB. Most patients (94%) with a CD4 T-lymphocyte count <350 cells/mm3 were associated with extrapulmonary TB (p <0.0001), whilst most patients (78%) with a CD4 T-lymphocyte count >350 cells/mm3 were associated with pulmonary TB (p = 0.0011). Eighty-two strains were pan-susceptible, four mono-resistant, four poly-resistant, two multidrug-resistant, and one was extensively drug-resistant. In the rifampicin-resistant strains, rpoB S531L was the mutation most frequently identified, whereas the inhA C15T and katG S315T1 mutations were present in isoniazid-resistant strains. The extensively drug-resistant strain also contained the mutation gyrA D94A. CONCLUSIONS: These data highlight the need to promptly diagnose the drug resistance of M. tuberculosis among all HIV-infected patients by systematically offering access to first- and second-line drug susceptibility testing and to tailor the treatment regimen based on the resistance patterns to reduce the number of deaths in HIV-infected patients.

Polyphenolic Compounds Nanostructurated with Gold Nanoparticles Enhance Wound Repair.

Gold nanoparticles (AuNPs) have been used in a wide range of applications, conferring to bio-molecules diverse properties such as delivery, stabilization, and reduction of the adverse effects of drugs or plant extracts. Polyphenolic compounds from Bacopa procumbens (B. procumbens) (BP) can modulate proliferation, adhesion, migration, and cell differentiation, reducing the artificial scratch area in fibroblast cultures and promoting wound healing in an in vivo model. Here, chemically synthesized AuNPs conjugated with BP (AuNP-BP) were characterized using UV-Vis, ATR-FTIR, DLS, zeta-potential, and TEM analysis. The results showed an overlap of the FTIR spectra of the polyphenolic compounds from B. procumbens adhered to the surface of the AuNPs. UV-vis analysis indicated that the average size of the AuNP-BP was 28 nm, while DLS analysis showed a size of 44.58 nm and, by TEM, a size of 16.5 nm with an icosahedral morphology was observed. These measurements suggest an increase in the size of the nanoparticles after conjugation with BP, compared to the sizes of 9 nm, 44.51 nm, and 14.17 nm for the unconjugated AuNPs, respectively. Furthermore, the zeta potential of the AuNPs, which was originally -36.3 ± 12.3 mV shifted to -18.2 ± 7.02 mV after conjugation with BP, indicating improved stability of the nanoparticles. Enhancement of the wound healing effect was evaluated by morphometric, histochemical, and FTIR changes in a rat wound excision model. Results showed that the nanoconjugation process reduced the BP concentrations by 100-fold to have the same wound healing effect as BP alone. Besides, histological and FTIR spectroscopy analyses demonstrated that AuNP-BP treatment exhibited better macroscopical performance, showing a reduction in inflammatory cells and an increased synthesis and improved organization of collagen fibers.

Dynamic response antibodies SARS-CoV-2 human saliva studied using two-dimensional correlation (2DCOS) infrared spectral analysis coupled with receiver operation characteristics analysis.

COVID-19 has affected the entire world due to the rapid spread of SARS-CoV-2, mainly through airborne particles from saliva, which, being easily obtained, help monitor the progression of the disease. Fourier transform infrared (FTIR) spectra combined with chemometric analysis could increase the diagnostic efficiency of the disease. However, two-dimensional correlation spectroscopy (2DCOS) is superior to conventional spectra as it helps to resolve the minute overlapped peaks. In this work, we aimed to use 2DCOS and receiver operating characteristic (ROC) analyses to compare the immune response in saliva associated with COVID-19, which could be important in biomedical diagnosis. FTIR spectra of human saliva samples from male (575) and female (366) patients ranging from 20 to 85 ± 2 years of age were used for the study. Age groups were segregated as G1 (20-40 ± 2 years), G2 (45-60 ± 2 years), and G3 (65-85 ± 2 years). The results of the 2DCOS analysis showed biomolecular changes in response to SARS-CoV-2. 2DCOS analyses of the male G1 + (1579,1644) and -(1531,1598) cross peaks evidenced changes such as amide I > IgG. Female G1 cross peaks -(1504,1645), (1504,1545) and -(1391,1645) resulted in amide I > IgG > IgM. The asynchronous spectra in 1300-900 cm-1 of the G2 male group showed that IgM is more important in diagnosing infections than IgA. Female G2 asynchronous spectra -(1027,1242) and + (1068,1176) showed that IgA > IgM is produced against SARS-CoV-2. The G3 male group evidenced antibody changes in IgG > IgM. The absence of IgM in the female G3 population diagnoses a specifically targeted immunoglobulin associated with sex. Moreover, ROC analysis showed sensitivity (85-89 % men; 81-88 % women) and specificity (90-93 % men; 78-92 % women) for the samples studied. The general classification performance (F1 score) of the studied samples is high for the male (88-91 %) and female (80-90 %) populations. This high PPV (positive predictive value) and NPV (negative predictive value) verify our segregation of COVID-19 positive and negative sample groups. Therefore, 2DCOS with ROC analysis using FTIR spectra have the potential for a non-invasive approach to monitoring COVID-19.

Implication of myddosome complex genetic variants in outcome severity of COVID-19 patients.

BACKGROUND/PURPOSE(S): During a viral infection, the immune response is mediated by the toll-like receptors and myeloid differentiation Factor 88 (MyD88) that play an important role sensing infections such as SARS-CoV-2 which has claimed the lives of more than 6.8 million people around the world. METHODS: We carried out a cross-sectional with a population of 618 SARS-CoV-2-positive unvaccinated subjects and further classified based on severity: 22% were mild, 34% were severe, 26% were critical, and 18% were deceased. Toll Like Receptor 7 (TLR7) single-nucleotide polymorphisms (rs3853839, rs179008, rs179009, and rs2302267) and MyD88 (rs7744) were genotyped using TaqMan OpenArray. The association of polymorphisms with disease outcomes was performed by logistic regression analysis adjusted by covariates. RESULTS: A significant association of rs3853839 and rs7744 of the TLR7 and MyD88 genes, respectively, was found with COVID-19 severity. The G/G genotype of the rs3853839 TLR7 was associated with the critical outcome showing an Odd Ratio = 1.98 (95% IC = 1.04-3.77). The results highlighted an association of the G allele of MyD88 gene with severe, critical and deceased outcomes. Furthermore, in the dominant model (AG + GG vs. AA), we observed an Odd Ratio = 1.70 (95% CI = 1.02-2.86) with severe, Odd Ratio = 1.82 (95% CI = 1.04-3.21) with critical, and Odd Ratio = 2.44 (95% CI = 1.21-4.9) with deceased outcomes. CONCLUSION: To our knowledge this work represents an innovative report that highlights the significant association of TLR7 and MyD88 gene polymorphisms with COVID-19 outcomes and the possible implication of the MyD88 variant with D-dimer and IFN-α concentrations.

Comparison of the Immune Response in Vaccinated People Positive and Negative to SARS-CoV-2 Employing FTIR Spectroscopy.

Various immunopathological events characterize the systemic acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Moreover, it has been reported that coronavirus disease 2019 (COVID-19) vaccination and infection by SARS-CoV-2 induce humoral immunity mediated by B-cell-derived antibodies and cellular immunity mediated by T cells and memory B cells. Immunoglobulins, cytokines, and chemokines play an important role in shaping immunity in response to infection and vaccination. Furthermore, different vaccines have been developed to prevent COVID-19. Therefore, this research aimed to analyze and compare Fourier-transform infrared (FTIR) spectra of vaccinated people with a positive (V-COVID-19 group) or negative (V-Healthy group) real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) test, evaluating the immunoglobulin and cytokine content as an immunological response through FTIR spectroscopy. Most individuals that integrated the V-Healthy group (88.1%) were asymptomatic; on the contrary, only 28% of the V-COVID-19 group was asymptomatic. Likewise, 68% of the V-COVID-19 group had at least one coexisting illness. Regarding the immunological response analyzed through FTIR spectroscopy, the V-COVID-19 group showed a greater immunoglobulins G, A, and M (IgG, IgA, and IgM) content, as well as the analyzed cytokines interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-ɑ), and interleukins 1ß, 6, and 10 (IL-1ß, IL-6, and IL-10). Therefore, we can state that it was possible to detect biochemical changes through FTIR spectroscopy associated with COVID-19 immune response in vaccinated people.

Detection of People Positive to COVID-19 through ATR-FTIR Spectra Analysis of Saliva using Machine Learning / Detección de Personas Positivas a COVID-19 mediante el Análisis de Espectros ATR-FTIR de Muestras de Saliva empleando Machine Learning

ABSTRACT COVID-19 is an infectious disease caused by the SARS-CoV-2 virus. This virus's spread is mainly through droplets released from the nose or mouth of an infected person. Although vaccines have been developed that effectively reduce the effects that this viral infection causes, the most effective method to contain the virus's spread is numerous tests to detect and isolate possible carriers. However, the response time, combined with the cost of actual tests, makes this option impractical. Herein, we compare some machine learning methodologies to propose a reliable strategy to detect people positive to COVID-19, analyzing saliva spectra obtained by Fourier transform infrared (FTIR) spectroscopy. After analyzing 1275 spectra, with 7 strategies commonly used in machine learning, we concluded that a multivariate linear regression model (MLMR) turns out to be the best option to identify possible infected persons. According to our results, the displacement observed in the region of the amide I of the spectrum, is fundamental and reliable to establish a border from the change in slope that causes this displacement that allows us to characterize the carriers of the virus. Being more agile and cheaper than reverse transcriptase polymerase chain reaction (RT-PCR), it could be reliably applied as a preliminary strategy to RT-PCR.

RESUMEN La COVID-19 es una enfermedad infecciosa ocasionada por el virus SARS-CoV-2. La propagación de este virus se produce principalmente a través de gotitas liberadas por la nariz o la boca de una persona infectada. Aunque se han desarrollado vacunas que permiten reducir efectivamente los efectos que esta infección viral provoca, el método más eficaz para contener la propagación del virus son las numerosas pruebas para detectar y aislar los posibles portadores. Sin embargo, el tiempo de respuesta, combinado con el costo de las pruebas reales, hace que esta opción sea poco práctica. Aquí, comparamos algunas metodologías de machine learning para proponer una estrategia confiable para detectar personas positivas a COVID-19 analizando espectros de saliva obtenidos por espectroscopia infrarroja transformada de Fourier (FTIR). Tras analizar 1275 espectros, con 7 estrategias comúnmente empleadas en el área de machine learning, concluimos que un modelo de regresión lineal multivariante (MLMR) resulta ser la mejor opción para identificar posibles infectados. De acuerdo con nuestros resultados, el desplazamiento observado en la región de la amida I del espectro, resulta fundamental y confiable para establecer una frontera a partir del cambio de pendiente que este provoca. Al ser más ágil y económica que la reacción en cadena de la polimerasa con transcriptasa inversa (RT-PCR), podría aplicarse confiablemente como estrategia preliminar a RT-PCR.

Artificial intelligence model validation before its application in clinical diagnosis assistance.

The process of selecting an artificial intelligence (AI) model to assist clinical diagnosis of a particular pathology and its validation tests is relevant since the values of accuracy, sensitivity and specificity may not reflect the behavior of the method in a real environment. Here, we provide helpful considerations to increase the success of using an AI model in clinical practice.

ACE and ACE2 Gene Variants Are Associated With Severe Outcomes of COVID-19 in Men.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the current coronavirus disease 2019 (COVID-19) pandemic, affecting more than 219 countries and causing the death of more than 5 million people worldwide. The genetic background represents a factor that predisposes the way the host responds to SARS-CoV-2 infection. In this sense, genetic variants of ACE and ACE2 could explain the observed interindividual variability to COVID-19 outcomes. In order to improve the understanding of how genetic variants of ACE and ACE2 are involved in the severity of COVID-19, we included a total of 481 individuals who showed clinical manifestations of COVID-19 and were diagnosed by reverse transcription PCR (RT-PCR). Genomic DNA was extracted from peripheral blood and saliva samples. ACE insertion/deletion polymorphism was evaluated by the high-resolution melting method; ACE single-nucleotide polymorphism (SNP) (rs4344) and ACE2 SNPs (rs2285666 and rs2074192) were genotyped using TaqMan probes. We assessed the association of ACE and ACE2 polymorphisms with disease severity using logistic regression analysis adjusted by age, sex, hypertension, type 2 diabetes, and obesity. The severity of the illness in our study population was divided as 31% mild, 26% severe, and 43% critical illness; additionally, 18% of individuals died, of whom 54% were male. Our results showed in the codominant model a contribution of ACE2 gene rs2285666 T/T genotype to critical outcome [odds ratio (OR) = 1.83; 95%CI = 1.01-3.29; p = 0.04] and to require oxygen supplementation (OR = 1.76; 95%CI = 1.01-3.04; p = 0.04), in addition to a strong association of the T allele of this variant to develop critical illness in male individuals (OR = 1.81; 95%CI = 1.10-2.98; p = 0.02). We suggest that the T allele of rs2285666 represents a risk factor for severe and critical outcomes of COVID-19, especially for men, regardless of age, hypertension, obesity, and type 2 diabetes.

Different kinds of stem cells in the development of SARS-CoV-2 treatments.

On February 11, 2020, the World Health Organization officially announced the coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), as an emerging recent pandemic illness, which currently has approximately taken the life of two million persons in more than 200 countries. Medical, clinical, and scientific efforts have focused on searching for new prevention and treatment strategies. Regenerative medicine and tissue engineering focused on using stem cells (SCs) have become a promising tool, and the regenerative and immunoregulatory capabilities of mesenchymal SCs (MSCs) and their exosomes have been demonstrated. Moreover, it has been essential to establishing models to reproduce the viral life cycle and mimic the pathology of COVID-19 to understand the virus's behavior. The fields of pluripotent SCs (PSCs), induced PSCs (iPSCs), and artificial iPSCs have been used for this purpose in the development of infection models or organoids. Nevertheless, some inconveniences have been declared in SC use; for example, it has been reported that SARS-CoV-2 enters human cells through the angiotensin-converting enzyme 2 receptor, which is highly expressed in MSCs, so it is important to continue investigating the employment of SCs in COVID-19, taking into consideration their advantages and disadvantages. In this review, we expose the use of different kinds of SCs and their derivatives for studying the SARS-CoV-2 behavior and develop treatments to counter COVID-19.

Astrocytes Are More Vulnerable than Neurons to Silicon Dioxide Nanoparticle Toxicity in Vitro.

Some studies have shown that silicon dioxide nanoparticles (SiO2-NPs) can reach different regions of the brain and cause toxicity; however, the consequences of SiO2-NPs exposure on the diverse brain cell lineages is limited. We aimed to investigate the neurotoxic effects of SiO2-NP (0-100 µg/mL) on rat astrocyte-rich cultures or neuron-rich cultures using scanning electron microscopy, Attenuated Total Reflection-Fourier Transform Infrared spectroscopy (ATR-FTIR), FTIR microspectroscopy mapping (IQ mapping), and cell viability tests. SiO2-NPs were amorphous particles and aggregated in saline and culture media. Both astrocytes and neurons treated with SiO2-NPs showed alterations in cell morphology and changes in the IR spectral regions corresponding to nucleic acids, proteins, and lipids. The analysis by the second derivative revealed a significant decrease in the signal of the amide I (α-helix, parallel ß-strand, and random coil) at the concentration of 10 µg/mL in astrocytes but not in neurons. IQ mapping confirmed changes in nucleic acids, proteins, and lipids in astrocytes; cell death was higher in astrocytes than in neurons (10-100 µg/mL). We conclude that astrocytes were more vulnerable than neurons to SiO2-NPs toxicity. Therefore, the evaluation of human exposure to SiO2-NPs and possible neurotoxic effects must be followed up.

Caracterización biomolecular mediante microespectroscopía FTIR en la fase de remodelación de la cicatrización en un modelo murino de lesión excisional / Biomolecular characterization by FTIR microspectroscopy in the modeling phase of wound cicatrization in a murine model of excisional injury

La cicatrización de la piel es un proceso complejo y organizado que involucra tres fases: inflamatoria, proliferativa y de remodelación. Es indispensable el análisis de este proceso biomolecularmente para investigar y proponer nuevas estrategias terapéuticas que mejoren la cicatrización o promuevan la regeneración. El objetivo de este proyecto fue analizar histológica y biomolecularmente mediante microespectroscopía infrarroja por transformada de Fourier (MFTIR) y su función de mapeo bioquímico, muestras de lesiones excisionales de piel, comparando los cambios morfológicos y espectroscópicos entre piel sana y piel cicatrizada. Se estandarizó un modelo de lesión excisional de piel en ratones hembra de la cepa NIH de 8 semanas de edad (n=16), provocando una herida excisional de 1 cm2. Se analizó piel sana (día 0) y cicatrizada (día 15 post-lesión) morfométrica, histológica y biomolecularmente mediante análisis fotográfico, técnica histológica y MFTIR con su función de mapeo. El análisis morfométrico demostró una reducción del área de la herida en un 87,6 % al día 15 post-lesión. Histológicamente, en la piel cicatrizada se evidenció un adelgazamiento de la epidermis y menor celularidad en la dermis, observándose la formación de tejido de granulación y fibras de colágena desorganizadas. Espectroscópicamente, se apreciaron cambios entre los dos grupos de estudio, principalmente en las bandas de lípidos y en la región de proteínas. El cálculo de las áreas bajo la curva y el mapeo bioquímico mostraron menor concentración de queratina y colágena en la piel cicatrizada, así como desorganización de las fibras de colágena. Se demostró la capacidad de la MFTIR para caracterizar de forma precisa los cambios biomoleculares en la cicatrización, entre ellos la cantidad de queratina, colágena, y el depósito y ordenamiento de las fibras de colágena asociadas a su maduración.

The skin cicatrization is a complex and organized process that involves three phases: inflammatory, proliferative, and remodeling. It is essential to analyze this process biomolecularly, in order to investigate and propose new therapeutic strategies that improve the healing or promote regeneration. The objective of this project was to analyze histological and biomolecularly through Fourier Transform infrared microspectroscopy (FTIRM) and its biochemical mapping function, samples of an excisional skin wound, comparing the morphological and spectroscopic changes between healthy skin and scarred skin. An excisional skin wound healing model was standardized using female, NIH strain 8-week-old mice (n = 16), provoking an excisional wound of 1 cm2. Healthy skin (day 0) and scarring skin (day 15 post-injury) were morphometrical, histological, and biomolecularly analyzed by digital picture analysis, histological technique, and FTIRM with its mapping function. The morphometric analysis showed a reduction of the wound area of 87.6 % at day 15 after wound. Histologically, in the scarred skin a thinning of the epidermis was evidenced, besides reduced cellularity in the dermis, granulation tissue formation, and disorganized collagen fibers were observed. Spectroscopically, changes between the study groups were appreciated, mainly in the lipid bands and in the protein region. The calculation of the areas under the curve and the biochemical mapping showed a lower concentration of keratin and collagen in the scarred skin, as well as collagen fibers disorganization. The ability of the FTIRM to accurately characterize biomolecular changes in cicatrization process was demonstrated, such as the amount of keratin, collagen, and the deposition and ordering of the collagen fibers associated with their maturation.

Biochemical similarity between cultured chondrocytes and in situ chondrocytes by chemometric analysis from FTIR microspectroscopy.

BACKGROUND AIMS: Fourier Transform Infrared Micro-spectroscopy (FTIRM) is an emerging tool that obtains images with biochemical information of samples that are too small to be chemically analyzed by conventional Fourier transform infrared (FTIR) spectroscopy techniques. So, the central objective of this project was to study the biochemical similarity between articular and cultured chondrocytes by chemometric analysis from FTIRM. METHODS: Nine samples of knee articular cartilage were obtained; each sample was divided into two fragments, one portion was used for FTIRM characterization in situ, and from another part, chondrocytes were obtained to be cultured (in vitro), which were subjected to an FTIRM to characterize their biomolecular components. The FTIRM spectra were normalized, and the second derivative was calculated. From these data, principal component analysis (PCA) and a chemometric comparison between in situ and cultured chondrocytes were carried out. Finally, the biochemical mapping was conducted obtaining micro-FTIR imaging. RESULTS: FTIRM spectra of in situ and in vitro chondrocytes were obtained, and different biomolecules were detected, highlighting lipids, proteins, glycosaminoglycans, collagen, and aggrecan. Despite slight differences in the FTIR spectra, the PCA proved the organic similarity between in situ chondrocytes and cultured chondrocytes, which was also observed in the analysis of the ratios related to the degradation of the articular cartilage and collagen. In the same way, the ability of the FTIRM to characterize the molecular biodistribution was demonstrated. CONCLUSION: The biochemical composition and biodistribution analysis using FTIRM have been useful for comparing cultured chondrocytes and in situ chondrocytes.

Characterization of the Biological Fingerprint and Identification of Associated Parameters in Stress Fractures by FTIR Spectroscopy.

INTRODUCTION: The stress fractures (SFs) are a common condition in athletes and military recruits, characterized by partial fracture caused by repetitive applications of stresses that are lower than the stress required to fracture the bone in a single loading. Fourier transform infrared (FTIR) spectroscopy gives information about the bone composition and also can determine the amount of a molecule. For this reason, the FTIR spectroscopy may be used as a tool for diagnosis of certain bone diseases related to the bone strength. In this research, we established the contributions of mineral and collagen properties to SF risk through FTIR spectroscopy, analyzing the biochemical profile differences between the healthy bone and the bone with an SF. MATERIALS AND METHODS: Previous written informed consent was obtained, and samples of the hip with an SF (n = 11) and healthy bone from the femur with traumatic fracture (n = 5) were obtained and analyzed employing FTIR spectroscopy and its biochemical mapping function. Then, using FTIR spectra and the map, the collagen content and ratios corresponding to matrix maturity, mineralization, carbonate substitution, acid phosphate substitution, and crystallinity were calculated. Moreover, a histopathological analysis through Masson's staining was conducted. RESULTS: The biochemical analysis showed that the bone with an SF presented a bone immaturity characterized by a higher content of collagen, lower matrix maturity, mineralization, carbonate and acid phosphate substitutions, and greater crystallinity compared to the healthy bone, being checked by the ratio analysis and biochemical mapping. Besides, Masson's stain showed a higher collagen content in the bone with an SF. CONCLUSIONS: The bone with an SF presented alterations in its biochemical composition, showing bone immaturity, which broadens the panorama of the condition to investigate future treatments or prophylactic techniques.

Revisión de las alteraciones en la expresión génica y vías apoptóticas provocadas en la nefrotoxicidad inducida por cisplatino / Review of alterations in gene expression and apoptotic pathways caused in nephrotoxicity induced by cisplatin

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Biochemical and histological effects of embryonic stem cells in a mouse model of renal failure.

Kidney diseases are a global public health problem. Despite significant advances in the understanding of renal failure (RF) and replacement therapies, this condition carries a series of complications and the life's quality of patients decreases. Differentiation capability of stem cells and their beneficial effects when they are implanted in animal models have been reported. Therefore, this work aimed to induce a long-term RF in mice, evaluating the biochemical and histological effects after implanting mouse embryonic stem cells (mESC). Mice were subjected to renal failure induction (RFI) employing cisplatin, subsequently received intraperitoneal (i.p.) injections of salt solution (control group, n=19) or 50 000 mESC (experimental group, n=19) at 24 hours, 7 days, and 13 days post-RFI. Ten animals in each group were used to analyze functional damage through serum biochemical analysis, and the mortality. For histopathological examination, three animals of each group were sacrificed at 5, 10, and 20 days post-RFI, analyzing the tubular system and glomeruli. Both groups showed blood urea nitrogen (BUN) and creatinine elevation three days post-RFI. Accumulated mortality was lower in the experimental group, presenting statistical significance. Respect to histopathological effects, the control group showed tubular dilatation, segmental focal glomerulosclerosis data, and collapsed glomeruli, while in the experimental group, glomerulosclerosis or collapsed glomeruli were not observed, evidencing regenerative data as characterized by large nuclei with prominent and binucleate nucleoli. In conclusion, mESC implant in mice with RFI significantly decreased the mortality, avoiding a greater histological deterioration related to the disease.

Nephroprotective Effect of Embryonic Stem Cells Reducing Lipid Peroxidation in Kidney Injury Induced by Cisplatin.

INTRODUCTION: The acute kidney injury (AKI) is characterized by a sudden glomerular filtration reduction. Renal or intrinsic causes of AKI include nephrotoxicity induced by exogenous agents like cisplatin, which causes oxidative stress altering the biochemical process and leading to apoptosis. Therefore, this research is aimed at analyzing the embryonic stem cells (ESC) nephroprotective effect in AKI induced by cisplatin, employing genetic, phenotypic, and microspectroscopic techniques. METHODS: Thirty mice were randomly divided into three groups (n = 10): the healthy, isotonic salt solution (ISS), and mouse embryonic stem cells (mESC) groups. The ISS and mESC groups were subjected to AKI using cisplatin; 24 h post-AKI received an intraperitoneal injection of ISS or 1 × 106 mESC, respectively. At days 4 and 8 post-AKI, five mice of each group were sacrificed to analyze the histopathological, genetic (PDK4 and HO-1), protein (p53), and vibrational microspectroscopic changes. RESULTS: Histopathologically, interstitial nephritis and acute tubular necrosis were observed; however, the mESC group showed a more preserved microarchitecture with high cellularity. Additionally, the PDK4 and HO-1 gene expression only increased in the ISS group on day 4 post-AKI. Likewise, p53 was more immunoexpressed at day 8 post-AKI in the ISS group. About biomolecular analysis by microspectroscopy, bands associated with lipids, proteins, and nucleic acids were evidenced. Besides, ratios related to membrane function (protein/lipid), unsaturated lipid content (olefinic/total lipid, olefinic/total CH2, and CH2/CH3), and lipid peroxidation demonstrated oxidative stress induction and lipid peroxidation increase mainly in the ISS group. Finally, the principal component analysis discriminated against each group; nonetheless, some data of the healthy and mESC groups at day 8 were correlated. CONCLUSIONS: The mESC implant diminishes cisplatin nephrotoxicity, once the protective effect in the reduction of lipid peroxidation was demonstrated, reflecting a functional and histological restoration.

Evaluation of the effect of heat damage on DNA extracted from the dental pulp of restored teeth.

We evaluate structural damage effects of heat on DNA obtained from the dental pulp of restored premolars. We studied three groups (A, B and C) each group comprised twenty premolars extracted from five patients. Three of the four premolars of each donator were restored with different materials: amalgam, glass ionomer and resin, and one unrestored premolar was used as control. The group A was not exposed to heat, while B and C groups were exposed to 100 °C and 300 °C, respectively. The DNA damage was evaluated as percentage of genotyping of 15 Short Tandem Repeats (STRs) and amelogenin and by Fourier-transform infrared spectroscopy (FTIR). The results showed 100% genotyping in all unheated premolars; however, the increase in heat decreased genotyping percentage due to DNA degradation. The amplifications from the premolars restored with glass ionomer and those unrestored were less affected, amplifying by approximately 80% at 300 °C. FTIR revealed that DNA structural damage occurred in the phosphate region, and changes in ribose were also shown; in addition, we detected presence of ß- three-calcium-phosphate (ß - TCP) due to heat treatment. Moreover, the phosphate region of DNA was a good indicator of DNA integrity related to the ratio of 1230/1085 cm-1 in the second derivative (asymmetric/symmetric PO2), which was major in premolars restored with glass ionomer heated at 100 °C, and this ratio is related to less DNA alterations and better genotyping; however this changes only were detected at 100 °C, suggesting that dental restoration with this material only protects dental pulp at temperatures below 300 °C.