Raman spectroscopy study of 7,8-dihydrofolate inhibition on the Wuhan strain SARS-CoV-2 binding to human ACE2 receptor.

Emerging evidence suggests that elevated levels of folic acid in the bloodstream may confer protection against Wuhan-SARS-CoV-2 infection and mitigate its associated symptoms. Notably, two comprehensive studies of COVID-19 patients in Israel and UK uncovered a remarkable trend, wherein individuals with heightened folic acid levels exhibited only mild symptoms and necessitated no ventilatory support. In parallel, research has underscored the potential connection between decreased folic acid levels and the severity of Covid-19 among hospitalized patients. Yet, the underlying mechanisms governing this intriguing inhibition remain elusive. In a quest to elucidate these mechanisms, we conducted a molecular dynamics simulation approach followed by a Raman spectroscopy study to delve into the intricate interplay between the folic acid metabolite, 7,8-dihydrofolate (DHF), and the angiotensin-converting enzyme ACE2 receptor, coupled with its interaction with the receptor-binding domain (RBD) of the Wuhan strain of SARS-CoV-2. Through a meticulous exploration, we scrutinized the transformation of the ACE2 + RBD complex, allowing these reactants to form bonds. This was juxtaposed with a similar investigation where ACE2 was initially permitted to react with DHF, followed by the exposure of the ACE2 + DHF complex to RBD. We find that DHF, when bonded to ACE2, functions as a physical barrier, effectively inhibiting the binding of the Wuhan strain RBD. This physicochemical process offers a cogent explanation for the observed inhibition of host cell infection in subjects receiving supplementary folic acid doses, as epidemiologically substantiated in multiple studies. This study not only sheds light on a potential avenue for mitigating SARS-CoV-2 infection but also underscores the crucial role of folic acid metabolites in host-virus interactions. This research paves the way for novel therapeutic strategies in the battle against COVID-19 and reinforces the significance of investigating the molecular mechanisms underlying the protective effects of folic acid in the context of viral infections.

Raman Spectroscopy and Machine-Learning for Early Detection of Bacterial Canker of Tomato: The Asymptomatic Disease Condition.

Bacterial canker of tomato is caused by Clavibacter michiganensis subsp. michiganensis (Cmm). The disease is highly destructive, because it produces latent asymptomatic infections that favor contagion rates. The present research aims consisted on the implementation of Raman spectroscopy (RS) and machine-learning spectral analysis as a method for the early disease detection. Raman spectra were obtained from infected asymptomatic tomato plants (BCTo) and healthy controls (HTo) with 785 nm excitation laser micro-Raman spectrometer. Spectral data were normalized and processed by principal component analysis (PCA), then the classifiers algorithms multilayer perceptron (PCA + MLP) and linear discriminant analysis (PCA + LDA) were implemented. Bacterial isolation and identification (16S rRNA gene sequencing) were realized of each plant studied. The Raman spectra obtained from tomato leaf samples of HTo and BCTo exhibited peaks associated to cellular components, and the most prominent vibrational bands were assigned to carbohydrates, carotenoids, chlorophyll, and phenolic compounds. Biochemical changes were also detectable in the Raman spectral patterns. Raman bands associated with triterpenoids and flavonoids compounds can be considered as indicators of Cmm infection during the asymptomatic stage. RS is an efficient, fast and reliable technology to differentiate the tomato health condition (BCTo or HTo). The analytical method showed high performance values of sensitivity, specificity and accuracy, among others.

Ag@ZnO/MWCNT ternary nanocomposite as an active and stable catalyst for the 4-nitrophenol reduction in water.

The nitroaromatic compounds, known as organic pollutants, have arising attention due to their carcinogenic character, highly dangerous to human health. In this work, the Ag@ZnO/MWCNT ternary nanocomposite synthesized via conjugation of sonochemical and solvothermal treatments manifests high performance in the reduction of 4-nitrophenol in the aqueous media (TOF value of 246 min-1µmol metal-1). The incorporation of MWCNT onto the nanocomposite structure favored the reusing of the catalysts even after eight consecutive catalytic runs without catalysts cleaning nor product removal. Obtained samples were characterized by XRD, TEM, UV-vis, Raman and FTIR spectroscopies. It was found that ultrasonic treatment at relatively moderate conditions leads to functionalization of MWCNT, the appearance of C=C and OH groups and change of electronic properties of Ag@ZnO/MWCNT composite which provide its stable material dispersion in aqueous solution and high catalytic performance in the 4-nitrophenol reduction. This technique may be effectively applied for the functionalization of carbon including materials for their usage in an aqueous media.

Frustules of Amphora sp. as a photonic crystal with photoluminescent CdS nanoparticles.

Diatom frustules have species-specific patterns of pores, striae, pores, and nanopores, periodically arranged on its silica surface, as sets of cavities that modify the vacuum electromagnetic density of states. Therefore, frustules may be considered photonic crystals; the interaction with light-emitting sources inside the pores may potentially result in enhancement or inhibition of their spontaneous radiative emission rate and frequencies. In this work, we studied the photoluminescence of cadmium sulfide nanoparticles (CdS-NP) deposited inside frustule cavities that conveyed evidence of cavity-NP interaction. We synthesized CdS-NP, a semiconductor compound achieving quantum dots small enough to impose confinement effects to the electronic states. CdS-NP and their clusters were physiosorbed onto the surface, striae, and predominantly inside the pores of the cleansed frustules of Amphora sp. A broad peak with a maximum intensity at 437 nm (2.84 eV) was recorded after excitation with a 375 nm light source, showing a large blue shift and signal amplification of the CdS-NP photoluminescence when these were embedded inside the pores of the silica frustule. Using the Brus equation, we estimated a NP size of 4.1 ± 0.2 nm for the CdS-NP snuggly packed inside the smaller pores of the frustule, of 10 ± 0.7 nm in average diameter, The emission Purcell enhancement factor for an emitting atom in a cavity was calculated. The obtained Q factor (c. 5) was smaller than typical Q factors for designed semiconductor cavities of similar dimensions, an expected situation if it is assumed that the pores are open-ended cavities.

TNF-α detection using gold nanoparticles as a surface-enhanced Raman spectroscopy substrate.

Background: TNF-α is a cytokine involved in inflammation. Surface-enhanced Raman spectroscopy (SERS) could be useful in its detection. Aim: Identify the TNF-α in an aqueous solution, using gold nanoparticles (AuNPs) as a SERS substrate. Materials & methods: Raman and SERS spectra were obtained from TNF-α samples, combined with AuNPs, with decreasing concentrations of TNF-α. The samples were analyzed using optical transmission spectroscopy, dynamic light scattering, and transmission electron microscopy. Results: Transmission electron microscopy/dynamic light scattering determined a change in the average diameter of the TNF-α/AuNPs (∼9.6 nm). Raman bands obtained were associated with aromatic amino acid side chains. We observe Raman signals for TNF-α concentrations as low as 0.125 pg/ml. Conclusion: TNF-α signal at physiological concentrations was determined with SERS.

Detection of SARS-CoV-2 and its S and N proteins using surface enhanced Raman spectroscopy.

The COVID-19 pandemic demonstrated the critical need for accurate and rapid testing for virus detection. This need has generated a high number of new testing methods aimed at replacing RT-PCR, which is the golden standard for testing. Most of the testing techniques are based on biochemistry methods and require chemicals that are often expensive and the supply might become scarce in a large crisis. In the present paper we suggest the use of methods based on physics that leverage novel nanomaterials. We demonstrate that using Surface Enhanced Raman Spectroscopy (SERS) of virion particles a very distinct spectroscopic signature of the SARS-CoV-2 virus can be obtained. We demonstrate that the spectra are mainly composed by signals from the spike (S) and nucleocapsid (N) proteins. It is believed that a clinical test using SERS can be developed. The test will be fast, inexpensive, and reliable. It is also clear that SERS can be used for analysis of structural changes on the S and N proteins. This will be an example of application of nanotechnology and properties of nanoparticles for health and social related matters.

Human Monocytic Ehrlichiosis, Mexico City, Mexico.

Little information is available about human infections by the members of the genus Ehrlichia in Mexico. Only 2 species, Ehrlichia canis and E. chaffensis, are known to cause disease in this country. We report a fatal case of human monocytic ehrlichiosis in Mexico City in a man who was homeless.

Determination of Salivary Sialic Acid Through Nanotechnology: A Useful Biomarker for the Screening of Breast Cancer.

PURPOSE: To demonstrate the usefulness of sialic acid (SA) in saliva as a biomarker for breast cancer (BC) and develop a new tool for early detection. METHODS: Considering that the amount of SA in human saliva is limited, the levels of SA were measured using surface-enhanced Raman spectroscopy (SERS) with tailored citrate-reduced silver nanoparticles. We calibrated the spectrum using analytical reagent SA. The 164 patients included in this study were undergoing screening mammography and/or ultrasound testing. The SA test was performed in the absence of previous information regarding the health of the subjects. Biopsies were performed to determine the diagnosis of cancer condition. The biopsy studies determined that 35 patients are BC affected and 129 gave negative results. RESULTS: SERS showed a sensitivity and specificity of 80 and 93%, respectively. The cut-off value for SA (12.5 mg/dL) was established through a receiver operating characteristic (ROC) curve analysis. The area under the curve of the ROC analysis resulted in 95% with this SA level cut-off. Our results suggest that SA may be a useful biomarker for the screening of breast cancer in women. CONCLUSIONS: Our results suggest that the SA levels measured from saliva may be highly sensitive and specific markers for the presence of breast cancer.

Determination of sialic acid levels by using surface-enhanced Raman spectroscopy in periodontitis and gingivitis.

OBJECTIVES: To compare the sialic acid (SA) levels in saliva among periodontitis-affected, gingivitis and control patients. METHODS: The study involved 93 subjects. The participants were divided into three groups: (1) 30 subjects without periodontal disease (control group); (2) 30 subjects with gingivitis; and (3) 33 subjects with periodontitis. The oral parameters examined were as follows: (a) Simplified Oral Hygiene Index; (b) Calculus Index; (c) Gingival Index; (d) probing pocket depth; and (e) level of epithelial attachment. SA levels in saliva were measured by means of surface-enhanced Raman spectroscopy (SERS). This method has demonstrated the capacity to detect extremely low concentrations of molecules. The spectrum was calibrated using analytical reagent SA. RESULTS: The obtained median values for SA concentrations were 5.98, 7.32, and 17.12 mg/dl for control, gingivitis, and periodontitis patients, respectively. CONCLUSIONS: Our measurements by SERS corroborate that in periodontitis-affected patients, the SA concentration is larger than their concentrations in either control or gingivitis patients. This confirms previous reports and opens the possibility of using SERS as a diagnostic tool.

Detection of Histamine Dihydrochloride at Low Concentrations Using Raman Spectroscopy Enhanced by Gold Nanostars Colloids.

In this paper, we report a fast and easy method to detect histamine dihydrochloride using gold nanostars in colloidal aqueous solution as a highly active SERS platform with potential applications in biomedicine and food science. This colloid was characterized with SEM and UV⁻Vis spectroscopy. Also, numerical calculations were performed to estimate the plasmonic resonance and electric field amplification of the gold nanoparticles to compare the difference between nanospheres and nanostars. Finally, aqueous solutions of histamine dihydrochloride were prepared in a wide range of concentrations and the colloid was added to carry out SERS. We found SERS amplified the Raman signal of histamine by an enhancement factor of 1 . 0 × 10 7 , demonstrating the capability of the method to detect low concentrations of this amine molecule.

Detection of Clavibacter michiganensis subsp. michiganensis Assisted by Micro-Raman Spectroscopy under Laboratory Conditions.

Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in México. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy (532 nm λ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants × 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and 1510 cm-1 were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

Determination of sialic acid in saliva by means of surface-enhanced Raman spectroscopy as a marker in adnexal mass patients: ovarian cancer vs benign cases.

BACKGROUND: To demonstrate the use of surface-enhanced Raman spectroscopy (SERS) to determine sialic acid (SA) levels in saliva using silver nanoparticles as substrates, in adnexal mass patients scheduled for surgical intervention to remove invasive masses, with the aim to compare SA levels in benign tumor vs ovarian cancer patients. METHODS: Quantification of SA levels was accomplished by measuring their SERS and calibrating with analytical reagent SA. The mean SA concentration in saliva from 37 benign adnexal mass resulted smaller (5.1 mg/dL) than the mean concentration in 15 Ovarium cancer patients (23 mg/dL). The cancer condition was determined by biopsy of the removed adnexal mass. The CA-125 biomarker was also measured. The predictive potential of both biomarkers is discussed, together with the malignity risk index (MRI). RESULTS: Our results showed a sensitivity/specificity of 80%/100% with a cutoff to distinguish between benign/cancer cases of SA 15.5 mg/dL, as established from a ROC analysis. Our results suggest that SA may be a more useful biomarker than CA-125 to detect ovarian cancer. CONCLUSIONS: Our results suggest that the SA levels measured from saliva may be as good predictors as the MRI index for the presence of ovarian cancer in sensitivity/negative predictive value and outperforms it in specificity/positive predictive value.

Zinc oxide decorated multi-walled carbon nanotubes: their bolometric properties.

We report the synthesis of MWNT/ZnO hybrid nanostructures. A simple, affordable, chemical procedure to functionalize MWNTs with ZnO nanoparticles was performed. A significant portion of the surface of MWNTs was covered with ZnO nanoparticles; these particles formed highly porous spherical nodules of 50-150 nm in diameter, sizes that are an order of magnitude larger than similar ZnO nanonodules reported in the literature. Hence, the self-assembled nanocomposite the ZnO exhibited a large surface-to-volume ratio, which is a very advantageous property for potential catalytic applications. The resultant MWNT/ZnO nanocomposites were characterized by x-ray diffraction, scanning and high-resolution transmission electron microscopy, and UV-vis and Raman spectroscopy. The temperature coefficient of resistance (TCR) of the nanocomposites was measured and reported. The average TCR value goes from -5.6%/K up to -18%/K, over temperature change intervals from 10 K to 1 K. Based on these TCR results, the nanocomposite MWNT/ZnO prepared in this work is a promising material, with potential application as a bolometric sensor.

Low cytotoxicity of anisotropic gold nanoparticles coated with lysine on peripheral blood mononuclear cells "in vitro".

The aim of this study was to evaluate the cytotoxic effects of anisotropic (non spherical morphologies) gold nanoparticles coated with the amino acid Lysine (Lys) on peripheral blood mononuclear cells (PBMC) "in vitro". Gold (Au) nanoparticles tested in this study were synthesized by a seed-mediated growth using Lys as a structure and shape directing agent. Cytotoxic effects were evaluated by cell viability (resazurin assay), reactive oxygen species (ROS) induction (2',7'-dichlorofluorescein diacetate assay), DNA damage (comet assay) and apoptosis/necrosis (AnnexinV/propidium iodide assay) after PBMC were exposed to increasing concentrations (10, 25, 50, 100, and 250µM) of AuNPs coated with Lys (AuNPs-Lys) at different exposure times (3, 6, 12, and 24h). The results demonstrated that AuNPs-Lys exhibited low cytotoxicity towards PBMC, (high cell viability), with low levels of ROS, DNA damage and apoptosis/necrosis detected after treatment. These data suggest that AuNPs-Lys, might be viable for biomedical application subject to further investigations.

Effect of graphene oxide on bacteria and peripheral blood mononuclear cells.

BACKGROUND: Driven by the potential biological applications of graphene, many groups have studied the response of cells exposed to graphene oxide (GO). In particular, investigations of bacteria indicate that there are 2 crucial parameters, which so far have only been investigated separately: GO size and exposure methodology. Our study took into account both parameters. We carefully characterized the samples to catalog sizes and structural properties, and tested different exposure methodologies: exposure in saline solution and in the presence of growth media. Furthermore, we performed experiments with peripheral blood mononuclear cells exposed to our GO materials. METHODS: Atomic force microscopy, scanning electron microscopy, Raman spectroscopy, X-ray photoelectron spectroscopy and transmission electron microscopy were used to characterize the morphology and composition of different samples of GO: GO-H2O, GO-PBS and GO-MG. Our samples had 2D sizes of ~100 nm (GO-H2O and GO-PBS) and >2 µm (GO-MG). We tested antibacterial activity and cytotoxicity toward peripheral blood mononuclear cells of 3 different GO samples. RESULTS: A size-dependent growth inhibition of Escherichia coli (DH5 α) in suspension was found, which proved that this effect depends strongly on the protocol followed for exposure. Hemocompatibility was confirmed by exposing peripheral blood mononuclear cells to materials for 24 hours; viability and apoptosis tests were also carried out. CONCLUSIONS: Our experiments provide vital information for future applications of GO in suspension. If its antibacterial properties are to be potentiated, care should be taken to select 2D sizes in the micrometer range, and exposure should not be carried out in the presence of grow media.

Raman Spectroscopy an Option for the Early Detection of Citrus Huanglongbing.

This research describes the application of portable field Raman spectroscopy combined with a statistical analysis of the resulting spectra, employing principal component analysis (PCA) and linear discriminant analysis (LDA), in which we determine that this method provides a high degree of reliability in the early detection of Huanglongbing (HLB) on Sweet Orange, disease caused by the bacteria Candidatus Liberibacter asiaticus. Symptomatic and asymptomatic plant samples of Sweet Orange (Citrus sinensis), Persian Lime (C. latifolia), and Mexican Lime (C. aurantifolia) trees were collected from several municipalities, three at Colima State and three at Jalisco State (HLB presence). In addition, Sweet Orange samples were taken from two other Mexican municipalities, one at San Luis Potosí and the other at Veracruz (HLB absent). All samples were analyzed by real-time PCR to determine its phytosanitary condition, and its spectral signatures were obtained with an ID-Raman mini. Spectral anomalies in orange trees HLB-positive, were identified in bands related to carbohydrates (905 cm(-1), 1043 cm(-1), 1127 cm(-1), 1208 cm(-1), 1370 cm(-1), 1272 cm(-1), 1340 cm(-1), and 1260-1280 cm(-1)), amino acids, proteins (815 cm(-1), 830 cm(-1), 852 cm(-1), 918 cm(-1), 926 cm(-1), 970 cm(-1), 1002 cm(-1), 1053 cm(-1), and 1446 cm(-1)), and lipids (1734 cm(-1), 1736 cm(-1), 1738 cm(-1), 1745 cm(-1), and 1746 cm(-1)). Moreover, PCA-LDA showed a sensitivity of 86.9 % (percentage of positives, which are correctly identified), a specificity of 91.4 % (percentage of negatives, which are correctly identified), and a precision of 89.2 % (the proportion of all tests that are correct) in discriminating between orange plants HLB-positive and healthy plants. The Raman spectroscopy technique permitted rapid diagnoses, was low-cost, simple, and practical to administer, and produced immediate results. These are essential features for phytosanitary epidemiological surveillance activities that may conduct a targeted selection of highly suspicious trees to undergo molecular DNA analysis.

Analysis of cytotoxic effects of silver nanoclusters on human peripheral blood mononuclear cells 'in vitro'.

The antimicrobial properties of silver nanoparticles (AgNPs) have made these particles one of the most used nanomaterials in consumer products. Therefore, an understanding of the interactions (unwanted toxicity) between nanoparticles and human cells is of significant interest. The aim of this study was to assess the in vitro cytotoxicity effects of silver nanoclusters (AgNC, < 2 nm diameter) on peripheral blood mononuclear cells (PBMC). Using flow cytometry and comet assay methods, we demonstrate that exposure of PBMC to AgNC induced intracellular reactive oxygen species (ROS) generation, DNA damage and apoptosis at 3, 6 and 12 h, with a dose-dependent response (0.1, 1, 3, 5 and 30 µg ml(-1)). Advanced electron microscopy imaging of complete and ultrathin-sections of PBMC confirmed the cytotoxic effects and cell damage caused by AgNC. The present study showed that AgNC produced without coating agents induced significant cytotoxic effects on PBMC owing to their high aspect ratio and active surface area, even at much lower concentrations (<1 µg ml(-1)) than those applied in previous studies, resembling what would occur under real exposure conditions to nanosilver-functionalized consumer products.

Manejo endodóntico de un primer premolar maxilar con tres raíces / Endodontic management of a three-rooted maxillary first premolar

Objetivo: informar el manejo endodóntico de un primer premolar maxilar con tres raíces, mediante tratamiento de conductos adecuado a estos casos. Caso clínico: un paciente de sexo femenino, de 25 años de edad, se presentó a la consulta con un diagnóstico de pulpitis irreversible asintomática en la pieza 14. A partir de los hallazgos clínicos y radiográficos iniciales -que hacían sospechar una alteración en el número de conductos-, se comprobó la presencia de tres conductos radiculares. Esta situación obligó a realizar el tratamiento endodóntico con cuidados específicos para localizar todos los conductos radiculares y prevenir desgastes excesivos y/o perforaciones. Conclusión: el diagnóstico oportuno de esta variación anatómica permite establecer modificaciones en el protocolo del tratamiento de conductos, a fin de evitar el debilitamiento excesivo o, incluso, la perforación de su estrecha anatomía radicular

Manejo endodóntico de un primer premolar maxilar con tres raíces / Endodontic management of a three-rooted maxillary first premolar

Objetivo: informar el manejo endodóntico de un primer premolar maxilar con tres raíces, mediante tratamiento de conductos adecuado a estos casos. Caso clínico: un paciente de sexo femenino, de 25 años de edad, se presentó a la consulta con un diagnóstico de pulpitis irreversible asintomática en la pieza 14. A partir de los hallazgos clínicos y radiográficos iniciales -que hacían sospechar una alteración en el número de conductos-, se comprobó la presencia de tres conductos radiculares. Esta situación obligó a realizar el tratamiento endodóntico con cuidados específicos para localizar todos los conductos radiculares y prevenir desgastes excesivos y/o perforaciones. Conclusión: el diagnóstico oportuno de esta variación anatómica permite establecer modificaciones en el protocolo del tratamiento de conductos, a fin de evitar el debilitamiento excesivo o, incluso, la perforación de su estrecha anatomía radicular (AU)