Perceptions of COVID-19 vaccines among osteopathic medical students (OMS).

BACKGROUND: Research has shown that physicians' recommendations are one of the top predictors for individuals to receive vaccines. This study examined the perceptions of new COVID-19 vaccines among the medical students at the X and the factors that influenced their opinions. OBJECTIVE: To measure X students' perception of a new COVID-19 vaccine and the factors which drive their opinions. METHODS: An electronic survey of 37 questions was distributed to Osteopathic Medical Students (OMS I-IV) of X in October of 2020. RESULTS: 1770 total students received the survey, and 197 responded (11%). 45% (88/197) of the respondents reported that they would receive new COVID-19 vaccines if they were available at the time of the survey, while 19% (37/197) reported that they had not yet decided. Confidence in the US healthcare system, pharmaceutical trust, the United States Food and Drug Administration's (FDA)'s minimum effectiveness level, adequate vaccine testing, additional vaccine dose, and antivaccine acquaintances were significant predictors of intended vaccine uptake. CONCLUSIONS: Our findings confirmed a low acceptance of the new COVID-19 vaccine among OMS students, which mirrored the general public's low acceptance rate. Better education of OMS about vaccination benefits and the vaccine development process may increase future immunization rates.

Ebselen oxide attenuates mechlorethamine dermatotoxicity in the mouse ear vesicant model.

Mechlorethamine (HN2) is an alkylating agent and sulfur mustard mimetic. Topical exposure to HN2 is associated with tissue blistering. Previous work in our laboratory has shown that ebselen (EB-1) possesses anti-vesicant, anti-inflammatory, anti-bacterial, anti-fungal, and cytoprotective properties, both in vivo and in vitro. We recently reported that ebselen oxide (EB-2), an analog of EB-1 with a tetravalent selenium atom, also possesses anti-bacterial and anti-fungal activity and confers cytoprotection against HN2 in vitro. The purpose of the present study was to determine the vesicant countermeasure potential of EB-2 using the mouse ear vesicant model (MEVM). Compared to control ears, mouse ears exposed to a single dose of HN2 (0.500 µmol/ear) showed an increase in wet weights, ear thickness, hyperplasia, vesication, and inflammatory cell infiltration after 24 h. Fluorescence microscopy of terminal deoxynucleotidyl transferase (TdT) dUTP nick end labeling (TUNEL)-stained sections showed that the occurrence of apoptosis extended from the epidermis of the HN2-treated side, all the way to the contralateral epidermis. In contrast, HN2-exposed ears treated topically with EB-2 at a test dose of 0.250 mg/ear showed a significant decrease in wet weight (12% less vs. HN2 alone), morphometric thickness (13% less vs. HN2 alone), and vesication. In addition, TUNEL staining revealed that HN2 ears treated with EB-2 (0.250 mg/ear) showed a decrease in apoptosis as compared to the HN2 group. EB-2 also reduced the abundance of matrix metalloproteinase-9 (MMP-9) in ear tissues exposed to HN2. Taken together, our study demonstrates that EB-2 is an efficacious countermeasure to HN2.

Complete genome sequence of T'Ho virus, a novel putative flavivirus from the Yucatan Peninsula of Mexico.

BACKGROUND: We previously reported the discovery of a novel, putative flavivirus designated T'Ho virus in Culex quinquefasciatus mosquitoes in the Yucatan Peninsula of Mexico. A 1358-nt region of the NS5 gene was amplified and sequenced but an isolate was not recovered. RESULTS: The complete genome of T'Ho virus was sequenced using a combination of unbiased high-throughput sequencing, 5' and 3' rapid amplification of cDNA ends, reverse transcription-polymerase chain reaction and Sanger sequencing. The genome contains a single open reading frame of 10,284 nt which is flanked by 5' and 3' untranslated regions of 97 and 556-nt, respectively. Genome sequence alignments revealed that T'Ho virus is most closely related to Rocio virus (67.4% nucleotide identity) and Ilheus virus (65.9%), both of which belong to the Ntaya group, followed by other Ntaya group viruses (58.8-63.3%) and Japanese encephalitis group viruses (62.0-63.7%). Phylogenetic inference is in agreement with these findings. CONCLUSIONS: This study furthers our understanding of flavivirus genetics, phylogeny and diagnostics. Because the two closest known relatives of T'Ho virus are human pathogens, T'Ho virus could be an unrecognized cause of human disease. It is therefore important that future studies investigate the public health significance of this virus.

Merida virus, a putative novel rhabdovirus discovered in Culex and Ochlerotatus spp. mosquitoes in the Yucatan Peninsula of Mexico.

Sequences corresponding to a putative, novel rhabdovirus [designated Merida virus (MERDV)] were initially detected in a pool of Culex quinquefasciatus collected in the Yucatan Peninsula of Mexico. The entire genome was sequenced, revealing 11 798 nt and five major ORFs, which encode the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and RNA-dependent RNA polymerase (L). The deduced amino acid sequences of the N, G and L proteins have no more than 24, 38 and 43 % identity, respectively, to the corresponding sequences of all other known rhabdoviruses, whereas those of the P and M proteins have no significant identity with any sequences in GenBank and their identity is only suggested based on their genome position. Using specific reverse transcription-PCR assays established from the genome sequence, 27 571 C. quinquefasciatus which had been sorted in 728 pools were screened to assess the prevalence of MERDV in nature and 25 pools were found positive. The minimal infection rate (calculated as the number of positive mosquito pools per 1000 mosquitoes tested) was 0.9, and similar for both females and males. Screening another 140 pools of 5484 mosquitoes belonging to four other genera identified positive pools of Ochlerotatus spp. mosquitoes, indicating that the host range is not restricted to C. quinquefasciatus. Attempts to isolate MERDV in C6/36 and Vero cells were unsuccessful. In summary, we provide evidence that a previously undescribed rhabdovirus occurs in mosquitoes in Mexico.

Selected ebselen analogs reduce mechlorethamine toxicity in vitro.

Sulfur mustard (SM) is a potent vesicant. The lack of an effective antidote makes SM a continued threat to both military and civilian settings. A surrogate agent, namely mechlorethamine (HN2), was used here to mimic the toxicity of SM, and the main objective of this study was to demonstrate if selected organoselenium analogs could protect cultured A-431 skin cells from HN2 toxicity. Test compounds included ebselen (EB-1) and three related organoselenium analogs (EB-2, EB-3 and EB-4). In the absence of test compound, a reproducible and robust cell death was observed in the cells following incubation with HN2 (25 µM, 24 or 48 h) while cells treated with test compound alone (15, 30 or 60 µM) for similar periods of time were generally not affected. When incubated in the presence of both HN2 and test compound for 24 or 48 h, it was found that EB-1, EB-2, EB-3 and EB-4 could spare the cells from death, with the EB-4 compound being the most effective at reducing HN2 toxicity. Light microscopy confirmed these findings. The organoseleniums were also examined for their effects on reducing lipid peroxidation in the A-431 skin cells. Among the test compounds, EB-4 reduced lipid peroxidation by HN2 to the greatest extent. These studies, taken together, validate that the organoselenium antioxidants tested here may serve a purpose in the discovery of medical countermeasures to vesicants.

Ebselen reduces the toxicity of mechlorethamine in A-431 cells via inhibition of apoptosis.

A series of test compounds were evaluated for an ability to reduce the toxicity of the nitrogen mustard mechlorethamine (HN2) in vitro. The test compounds included resveratrol, pterostilbene, vitamin C, ebselen, ebselen diselenide, and ebselen-sulfur. Among them, ebselen demonstrated the highest degree of protection against HN2 toxicity. To this end, pretreatment of the cells with ebselen offered protection against the toxicant whereas no protection was observed when cells were first incubated with HN2 and then treated with ebselen. Significant increases in caspase 3 and caspase 9 activities were observed in response to HN2, and ebselen was found to reduce these effects. Taken together, the data presented here indicate that ebselen is an effective countermeasure to nitrogen mustard in vitro, which is worthy of future investigation in vivo.

Orthobunyavirus antibodies in humans, Yucatan Peninsula, Mexico.

We performed a serologic investigation to determine whether orthobunyaviruses commonly infect humans in the Yucatan Peninsula of Mexico. Orthobunyavirus-specific antibodies were detected by plaque reduction neutralization test in 146 (18%) of 823 persons tested. Further studies are needed to determine health risks for humans from this potentially deadly group of viruses.

Identification of a novel subtype of South River virus (family Bunyaviridae).

We previously reported the isolation of South River virus (SORV) from a pool of mosquitoes collected in the Yucatan Peninsula of Mexico (Farfan-Ale et al. in Vector Borne Zoonotic Dis 10:777-783, 5). The isolate (designated SORV-252) was identified as SORV after a 197-nucleotide region of its small RNA genome segment was sequenced. In the present study, the complete small and medium RNA genome segments and part of the large RNA genome segment of SORV-252 were sequenced and shown to have 92%, 85% and 90% nucleotide sequence identity, respectively, to the homologous regions of the prototype SORV isolate (NJO-94F). To determine the antigenic relationship between SORV-252 and NJO-94F, cross-plaque reduction neutralization tests (PRNTs) were performed using sera from mice inoculated with these viruses. SORV-252 and NJO-94F were distinguishable in the cross-neutralization assays; there was a twofold difference in the PRNT titers in one direction and a fourfold difference in the other direction, suggesting that SORV-252 represents a novel subtype of SORV. Additionally, SORV-252 and NJO-94F have distinct plaque morphologies in African green monkey kidney (Vero) cells. In conclusion, we provide evidence that a novel subtype of SORV is present in the Yucatan Peninsula of Mexico.

Sequence and phylogenetic data indicate that an orthobunyavirus recently detected in the Yucatan Peninsula of Mexico is a novel reassortant of Potosi and Cache Valley viruses.

We determined the complete nucleotide sequences of the small (S) and medium (M) RNA segments of an orthobunyavirus isolated from mosquitoes in the Yucatan Peninsula of Mexico. A 528-nt region of the large (L) RNA segment was also sequenced. The S RNA segment has greatest nucleotide identity to the homologous region of Cache Valley virus (CVV; 98%) followed by Potosi virus (POTV; 89%) and Northway virus (86%). The M RNA segment has 96% nucleotide identity to the homologous region of POTV, and less than 74% nucleotide identity to the homologous regions of all other orthobunyaviruses for which M segment sequence data are available. The L RNA segment has greatest nucleotide identity to the homologous region of POTV (98%) followed by CVV (82%) and Tensaw virus (77%). These data indicate that the virus, tentatively named Cholul virus (CHLV), is a novel reassortant that acquired its S RNA segment from CVV and its M and L RNA segments from POTV. Phylogenetic data support this conclusion.

Nucleotide sequencing and serologic analysis of Cache Valley virus isolates from the Yucatan Peninsula of Mexico.

Nucleotide sequencing was performed on part of the medium and large genome segments of 17 Cache Valley virus (CVV) isolates from the Yucatan Peninsula of Mexico. Alignment of these sequences to all other sequences in the Genbank database revealed that they have greatest nucleotide identity (97-98 %) with the equivalent regions of Tlacotalpan virus (TLAV), which is considered to be a variety of CVV. Next, cross-plaque reduction neutralization tests (PRNTs) were performed using sera from mice that had been inoculated with a representative isolate from the Yucatan Peninsula (CVV-478) or the prototype TLAV isolate (61-D-240). The PRNT titers exhibited a twofold difference in one direction and no difference in the other direction suggesting that CVV-478 and 61-D-240 belong to the same CVV subtype. In conclusion, we demonstrate that the CVV isolates from the Yucatan Peninsula of Mexico are genetically and antigenically similar to the prototype TLAV isolate.

[Hospital-acquired anemia and decrease of hemoglobin levels in hospitalized patients]. / Anemia intrahospitalaria y descenso de hemoglobina en pacientes internados.

It is common to observe the development of anemia in hospitalized patients, especially in critical cases. Few studies have evaluated its prevalence and associated factors in patients in the general ward. The purpose of this study is to determine the prevalence, characteristics and associated clinical factors of hospital-acquired anemia and the drop of hemoglobin concentration in hospitalized patients. This is a cross-sectional, prospective and descriptive study. A total of 192 consecutive in-patients in the general ward were studied. Associated risk factors to the drop in hemoglobin by ≥ 2g/dl were analyzed; 139 patients (72.4%) presented anemia; 89 of them (46.4%) had it at admission and 50 (26%) developed hospital-acquired anemia, 47 out of 192 showed a drop in hemoglobin ≥ 2 g/dl(24.48%). They also presented lower values of hematocrite and hemoglobin at discharge (p = 0.01), parenteral hydration at a higher volume (p = 0.01), and lengthier hospitalizations (p = 0.0001). In the univariate analysis, the following variables were statistically significant risk factors: leukocytosis ≥ 11000 mm3 (OR; IC95%: 2,02; 1.03-4; p = 0.01), hospitalization days ≥ 7 (OR; IC95%:3.39; 1.62-7.09; p = 0.0006), parenteral hydration ≥ 1500 ml/day (OR; IC95%: 2.47; 1.06-6.4; p = 0.01), central venous access (OR; IC95%:10.29; 1.75-108.07; p = 0.003) and hospital-acquired anemia (OR; IC95%: 7.06; 3.41-15.83; p = 0.00000004). In the multivariate analysis, the following variables were independent predictive factors of the hemoglobin decrease = 2 g/dl: leukocytosis ≥ 11000 mm3 (OR; IC95%: 2.45; 1.14-5,27; p = 0.02), hospitalization days ≥ 7 (OR; IC95%:5.15; 2.19-12.07; p = 0.0002), parenteral hydration ≥ 1500 ml/day (OR; IC95%: 2.95; 1.13-7.72; p = 0.02), central venous access (OR; IC95%:8.82; 1.37-56.82; p = 0.02). Hospital-acquired anemia has a high prevalence. Lengthier stays, presence of leukocytosis, parenteral hydration and central venous access placement are predictive factors of the drop in hemoglobin ≥ 2 g/dl.

Isolation and sequence analysis of Culex flavivirus from Culex interrogator and Culex quinquefasciatus in the Yucatan Peninsula of Mexico.

Previously, we reported a high prevalence of Culex flavivirus (CxFV) in Culex quinquefasciatus (Say) in the Yucatan Peninsula of Mexico. To determine whether other Culex spp. mosquitoes in this region are susceptible to natural CxFV infection, Cx. bahamensis (Dyar and Knab), Cx. coronator (Dyar and Knab), Cx. interrogator (Dyar and Knab), Cx. nigripalpus (Theobald) and Cx. opisthopus (Komp) in the Yucatan Peninsula of Mexico were tested for CxFV. Two pools of Cx. interrogator were positive. The envelope protein genes of these isolates and 16 isolates from Cx. quinquefasciatus were sequenced and shown to have > or =99.2% nucleotide identity. These data suggest that there is limited genetic diversity among CxFV isolates in the Yucatan Peninsula of Mexico.

Host-feeding preference of the mosquito, Culex quinquefasciatus, in Yucatan State, Mexico.

Studies were conducted to determine the host-feeding preference of Culex quinquefasciatus Say (Diptera: Culicidae) in relation to the availability of human and domestic animals in the city of Merida, Yucatan State, Mexico. Mosquitoes were collected in the backyards of houses using resting wooden boxes. Collections were made five times per week from January to December 2005. DNA was extracted from engorged females and tested by PCR using universal avian- and mammalian-specific primers. DNA extracted from avian-derived blood was further analyzed by PCR using primers that differentiate among the birds of three avian orders: Passeriformes, Columbiformes and Galliformes. PCR products obtained from mammalian-derived blood were subjected to restriction enzyme digestion to differentiate between human-, dog-, cat-, pig-, and horse-derived blood meals. Overall, 82% of engorged mosquitoes had fed on birds, and 18% had fed on mammals. The most frequent vertebrate hosts were Galliformes (47.1%), Passeriformes (23.8%), Columbiformes (11.2%) birds, and dogs (8.8%). The overall human blood index was 6.7%. The overall forage ratio for humans was 0.1, indicating that humans were not a preferred host for Cx. quinquefasciatus in Merida.

Universal primers for the amplification and sequence analysis of actin-1 from diverse mosquito species.

We report the development of universal primers for the reverse-transcription polymerase chain reaction (RT-PCR) amplification and nucleotide sequence analysis of actin cDNAs from taxonomically diverse mosquito species. Primers specific to conserved regions of the invertebrate actin-1 gene were designed after actin cDNA sequences of Anopheles gambiae, Bombyx mori, Drosophila melanogaster, and Caenorhabditis elegans. The efficacy of these primers was determined by RT-PCR with the use of total RNA from mosquitoes belonging to 30 species and 8 genera (Aedes, Anopheles, Culex, Deinocerites, Mansonia, Psorophora, Toxorhynchites, and Wyeomyia). The RT-PCR products were sequenced, and sequence data were used to design additional primers. One primer pair, denoted as Act-2F (5'-ATGGTCGGYATGGGNCAGAAGGACTC-3') and Act-8R (5'-GATTCCATACCCAGGAAGGADGG-3'), successfully amplified an RT-PCR product of the expected size (683-nt) in all mosquito spp. tested. We propose that this primer pair can be used as an internal control to test the quality of RNA from mosquitoes collected in vector surveillance studies. These primers can also be used in molecular experiments in which the detection, amplification or silencing of a ubiquitously expressed mosquito housekeeping gene is necessary. Sequence and phylogenetic data are also presented in this report.

What Physicians Wished They Would Have Learned in Medical School: a Survey.

PURPOSE: Medical students must be provided the basic science knowledge appropriate and applicable for preparing them for best-practice medicine. To date, there have been no documented studies in the USA that have directly surveyed practicing physicians on their perspectives of their basic science/preclinical medical school education and how it could be modified to help them deliver best patient care. This study was the first to examine this information. METHOD: A survey was administered to the alumni of Touro College of Osteopathic Medicine, Harlem, NY (2011-2018), with questions on examining perspectives on basic science disciplines, the need for a basic science refresher course, and other educational topics. In addition, questions relating to demographics and type of medical practice were also asked. Statistical analysis was performed using SPSS. RESULTS: (1) Gender (N = 122): 55% male and 44% female; (2) medical specialty (N = 107): 51.40% Primary Care physicians (Family medicine, Internal medicine, Pediatrics), 48.60% Other Specialties; (3) top Disciplines that "should have more": Physiology (41.1%), Pharmacology (39.3%), and Preventative Medicine/Public Health (39.3%); Top disciplines that "should have less": Histology Laboratory (38.32%), Embryology (35.51%), Histology (didactic) (28.30%) (N = 107); (4) top topics "most important" to be included in curriculum: Analysis of Journal Articles (70.10%), Clinical Cases (70.1%), and Early Patient Exposure (64.5%) (N = 107); (5) presentation of a clinically relevant Basic Science refresher course had a positive response (84.4%) (N = 107). CONCLUSIONS: Pharmacology, Physiology, Clinical Cases, Journal Article Analysis, and Early Patient exposure were among topics requiring "more" in preclinical education. A clinically relevant basic science course was deemed useful. The perspectives of practicing physicians should be included when designing future medical school curriculums.

Are Medical Students Prepared to Model Healthy Behaviors for Their Future Patients? A Survey Comparing Aged-Matched Peers.

BACKGROUND: Osteopathic medical students (OMS) who establish healthy behaviors for themselves are more likely to counsel their future patients on appropriate self-care. This study compared the lifestyle habits of OMS with those of age-matched peers in other areas of study, which served as the control group. METHOD: In the fall of 2018, a survey was administered to OMS of the New York Institute of Technology College of Osteopathic Medicine (NYIT-COM) (group I) and graduate programs from the same school (group II), to assess their lifestyle habits. Questions on demographics were additionally included. RESULTS: There were 398 total responses: 83.2% (N = 331) from group I and 16.9% (N = 67) from group II, with 25 being the mean age of the respondents. Group I (53.2%) reported to studying at least 5-10 h per day, while 20.1% reported to studying more than 10 h. Group II reported 37.3% and 9.0%, respectively, of study time. Group I exercised more times per week (2-3 times) than group II and for a longer duration (30-60 min). Group I slept more than group II (6-8 h), yet reported to using more substances to stay awake. CONCLUSIONS: OMS studied, exercised, and slept more than age-matched peers, but used more substances to stay awake. Aspects of this study are encouraging, but suggest that further evaluation is needed for schools to assist students establish lifelong habits to encourage the wellness of their future patients.

Complete nucleotide sequences of the small and medium RNA genome segments of Kairi virus (family Bunyaviridae).

We determined the complete nucleotide sequences of the small (S) and medium (M) RNA genome segments of a Kairi virus (KRIV) isolate from the Yucatan Peninsula of Mexico. The S segment consists of 992 nucleotides, and the M segment consists of 4,619 nucleotides. Phylogenetic analyses were conducted on each genomic segment, and these data are discussed. A 526 nucleotide region of the large (L) segment was also sequenced. This is the first study to present sequence and phylogenetic data for a KRIV isolate from Latin America.

Analysis of a set of missense, frameshift, and in-frame deletion variants of BRCA1.

Germline mutations that inactivate BRCA1 are responsible for breast and ovarian cancer susceptibility. One possible outcome of genetic testing for BRCA1 is the finding of a genetic variant of uncertain significance for which there is no information regarding its cancer association. This outcome leads to problems in risk assessment, counseling and preventive care. The purpose of the present study was to functionally evaluate seven unclassified variants of BRCA1 including a genomic deletion that leads to the in-frame loss of exons 16/17 (Delta exons 16/17) in the mRNA, an insertion that leads to a frameshift and an extended carboxy-terminus (5673insC), and five missense variants (K1487R, S1613C, M1652I, Q1826H and V1833M). We analyzed the variants using a functional assay based on the transcription activation property of BRCA1 combined with supervised learning computational models. Functional analysis indicated that variants S1613C, Q1826H, and M1652I are likely to be neutral, whereas variants V1833M, Delta exons 16/17, and 5673insC are likely to represent deleterious variants. In agreement with the functional analysis, the results of the computational analysis also indicated that the latter three variants are likely to be deleterious. Taken together, a combined approach of functional and bioinformatics analysis, plus structural modeling, can be utilized to obtain valuable information pertaining to the effect of a rare variant on the structure and function of BRCA1. Such information can, in turn, aid in the classification of BRCA1 variants for which there is a lack of genetic information needed to provide reliable risk assessment.

Gonotrophic cycle estimate for Culex quinquefasciatus in Mérida, Yucatán, México.

West Nile virus (WNV) has been present in the Yucatán State, México, since 2002. Culex quinquefasciatus, one of the main vectors of WNV transmission in the United States, is also common in Mexico and may be a key vector of WNV transmission t o humans in t he Yucatán. The aim of this study was to determine the length of the gonotrophic cycle and the survival rates of Cx. quinquefasciatus from Mérida, Yucatán, during the rainy versus the dry season. Mosquitoes were collected during 25-day periods in October (rainy season) and in April (dry season), and captured females were classified by abdominal appearance (freshly fed, late-stage fed, half gravid, and subgravid). To determine the age structure as nulliparous and parous females and to calculate the gonotrophic cycle through a time series and the mosquito survival, we used Davidson formulae. Also, vitellogenesis analysis to monitor egg maturity was conducted during both seasons. Cross-correlation data suggested a similar length of the gonotrophic cycle (4 days) in both seasons. Oogenic development required a minimum of 72 h in each season. However, survival of the mosquito population collected in the rainy season was significantly higher (0.91) with a mean temperature of 28 +/- 1.57 degrees C than was survival in the dry season (0.78) with a mean temperature of 29 +/- 1.10 degrees C. Survival, although higher during the rainy season, did not influence the length of the gonotrophic cycle of Cx. quinquefasciatus in Yucatán.

Reduction of vesicant toxicity by butylated hydroxyanisole in A-431 skin cells.

Mechlorethamine (HN2) and 2-chloroethylethyl sulfide (CEES) are mustard vesicants. Butylated hydroxyanisole (BHA) is a phenolic antioxidant. In the present work, the ability of BHA to reduce the toxicity of HN2 and CEES was investigated using A-431 skin cells. HN2 toxicity was found to be dependent, at least in part, on the cellular glutathione (GSH) status. BHA also decreased HN2-induced DNA damage and lipid peroxidation. The cytoprotective activity of BHA was also compared with that of the structurally unrelated antioxidant ebselen. Whereas ebselen (30 microM) protected skin cells from the toxicity of both HN2 (10-80 microM) and CEES (10-40 microM), BHA (100 microM) reduced the toxicity of HN2 only. Taken together, these data suggest that antioxidants such as BHA or ebselen may serve as useful treatments for injury caused by blistering agents.