Characterization and expression of domains of Alphaherpesvirus bovine 1/5 envelope glycoproteins B in Komagataella phaffi.

BACKGROUND: Bovine herpes virus (BoHV 1 and BoHV-5) are the causative agents of infectious bovine rhinotracheitis (IBR). IBR is responsible for important economic losses in the cattle industry. The envelope glycoprotein B (gB) is essential for BoHV infection of cattle's upper respiratory and genital tract. gB is one of the main candidate antigens for a potential recombinant vaccine since it induces a strong and persistent immune response. RESULTS: In this study, gB of BoHV-1 and BoHV-5 was characterized in terms of function, structure, and antigenicity through bioinformatics tools. gB showed conserved sequence and structure, so, both domains named PH Like 1 and 2 domains of each virus were selected for the design of a bivalent vaccine candidate. The immunoinformatic study showed that these two domains have epitopes recognizable by B and T lymphocytes, followed by this, the cDNA domains from BoHV-1/5 gB (Domains-gB) were transformed into the yeast Komagataella phaffii GS115 (previously known as Pichia pastoris). A recombinant protein with molecular weight of about 110 kDa was obtained from the culture media. The vaccine candidate protein (Domains-gB) was recognized by a monoclonal antibody from a commercial ELISA kit used for IBR diagnostic, which may suggest that the epitopes are conserved of the entire infectious virus. CONCLUSION: Overall, it was shown that the recombinant domains of BoHV-1/5 gB have antigenic and immunogenic properties similar to the native gB. This vaccine candidate is promising to be used in future studies to assess its immunogenicity in an animal model.

Evidence of bovine leukemia virus circulating in sheep and buffaloes in Colombia: insights into multispecies infection.

Bovine leukemia virus (BLV) is the causative agent of leukemia/lymphoma in cattle. However, previous evidence has shown its presence in other species of livestock as well as in humans, suggesting that other species can be accidental hosts of the virus. In viral infections, receptors that are common to different animal species are proposed to be involved in cross-species infections. For BLV, AP3D1 has been proposed to be its receptor, and this protein is conserved in most mammalian species. In Colombia, BLV has been reported in cattle with high prevalence rates, but there has been no evidence of BLV infections in other animal species. In this study, we tested for the virus in sheep (n = 44) and buffaloes (n = 61) from different regions of Colombia by nested PCR, using peripheral blood samples collected from the animals. BLV was found in 25.7% of the animals tested (12 buffaloes and 15 sheep), and the results were confirmed by Sanger sequencing. In addition, to gain more information about the capacity of the virus to infect these species, the predicted interactions of AP3D1 of sheep and buffaloes with the BLV-gp51 protein were analyzed in silico. Conserved amino acids in the binding domains of the proteins were identified. The detection of BLV in sheep and buffaloes suggests circulation of the virus in multiple species, which could be involved in dissemination of the virus in mixed livestock production settings. Due to the presence of the virus in multiple species and the high prevalence rates observed, integrated prevention and control strategies in the livestock industry should be considered to decrease the spread of BLV.

High prevalence of persistently infected animals from bovine viral diarrhea in Colombian cattle.

BACKGROUND: Bovine Viral Diarrhea Virus (BVDV) is associated with gastrointestinal, respiratory and reproductive diseases of livestock across the world that causes continuous economic losses in the cattle industry. This virus can establish a persistent infection (PI) in calves after the fetal infection, making BVDV positive catle carriers and primary reservoirs which will constantly transmit the virus to healthy and new-born animals. For this reason, the detection of the PI animals in herds is the first line of prevention of the viral infection. RESULTS: In this study, PI animals were detected in five different regions of Colombia through RT-PCR techniques and confirmed by sequencing. BVDV genotypes were determined using one fragment of the 5'UTR. It was found a 7% BVDV prevalence in animals and 22% in farms; and genotype 1 was identified as a single genotype for all of the samples. All samples were BVDV 1a. CONCLUSION: This is the first report in Colombia with higher prevalence rates compared with other places in the world, turned out to be of great importance for the ranchers, the vaccine producers and animal health control parties.

Characterization of fecal microbiota of children with diarrhea in 2 locations in Colombia.

OBJECTIVES: Diarrhea is a leading cause of mortality and morbidity in children younger than 5 years in impoverished regions of the world. Our aim was to compare the fecal microbiota of healthy children with that of children with clinical diarrhea in a population from a tropical highland in Colombia, South America. Our hypothesis was that a reduced prevalence of inherent Lactobacillus and Bifidobacterium species would be associated with enteric viral and bacterial pathogens. METHODS: Children between 1 and 5 years of age from 2 different locations were evaluated for presence of clinical diarrhea. Nucleic acid, isolated from fecal samples, was used to determine by molecular protocols the abundance of inherent bacterial species and presence of enteric pathogens compared with clinically healthy children. The effect of host demographic factors on incidence of diarrhea was also analyzed. RESULTS: : The composition of the fecal microbiota was affected by host demographic factors: age, health status, location, and sex. In partial support of our hypothesis, the relative abundance of commensal Bifidobacterium and Lactobacillus species was inversely correlated with incidence of diarrhea regardless of location. CONCLUSIONS: Our results suggested that changes in fecal microbiota composition of children with clinical diarrhea are associated with certain demographic factors that should be considered before designing a prophylactic intervention. Delivery of certain Lactobacillus species and Bifidobacterium species or a diet rich in bifidogenic components that promote growth of Bifidobacterium species could provide a prophylactic effect to ameliorate the effect of diarrhea in children at risk.

Humoral Immune Response of Mice against a Vaccine Candidate Composed of a Chimera of gB of Bovine Alphaherpesviruses 1 and 5.

Infectious bovine rhinotracheitis (IBR) and bovine meningoencephalitis are caused by Bovine alphaherpesvirus (BoHV) types 1 and 5, which seriously threaten the global cattle industry. Vaccination to improve immunity is the most direct and effective means to prevent these conditions. Glycoprotein B (gB) is essential for the attachment of both viruses to permissive cells, and is a major target of the host immune system, inducing a strong humoral response. The aim of this study was to evaluate, in a murine model, the immune response of a candidate vaccine formulation composed of a chimeric BoHV-1 and BoHV-5 gB (DgB), expressed in Komagataella phaffii. The chimeric DgB vaccine adjuvanted with Montanide 50 ISA V2 or aluminum hydroxide was administered intramuscularly or subcutaneously. A control group and a group that received a commercial vaccine were inoculated subcutaneously. Higher titers of neutralizing antibodies against BoHV-1, BoHV-5, and a natural BoHV-1/5 recombinant strain were obtained with the oil-based candidate vaccine formulation administered intramuscularly. The results demonstrated that the chimeric DgB conserved important epitopes that were able to stimulate a humoral immune response capable of neutralizing BoHV-1, BoHV-5, and the recombinant strain, suggesting that the vaccine antigen is a promising candidate to be further evaluated in cattle.

Letter Spacing Does not Affect Memory and Metamemory.

Some perceptual manipulations, such as font size or bolding, can affect the estimations of future memorability (i.e., judgments of learning or JOLs). In two experiments, we studied the effect on JOLs of another perceptual manipulation: the interletter spacing. Spacing may decrease JOLs via beliefs on the effect of spacing on memory, or it may increase JOLs via feelings of processing fluency. In Experiment 1, we found that people do not hold any particular belief on the effect of spacing on memory for a list of words. In Experiment 2, we found that letter spacing did not affect JOLs. We also replicated the results that participants believe that words in large font size will be better remembered and that they rate words in large font size with higher JOLs. In sum, this research showed that not all the perceptual characteristics are used as cues to metamemory.

Potentially Inappropriate Use of Opioids in the Management of Migraine in Colombia.

RESEARCH OBJECTIVE: To identify the frequency of opioid use in a group of patients diagnosed with migraine in Colombia. METHODS: Study of a retrospective cohort of patients with a diagnosis of migraine and a first prescription of antimigraine drugs from emergency services and a priority outpatient clinic. Sociodemographic, clinical, and pharmacological variables were identified; a 12-month follow-up was carried out to identify the use of a new opioid. RESULTS: A total of 6309 patients with a diagnosis of migraine were identified, with a mean age of 35.5 ± 12.3 years, of which 81.3% were women. Nonsteroidal anti-inflammatory drugs (51.1%) were the most frequently prescribed medications, followed by ergotamine + caffeine (31.3%), acetaminophen (15.05%), and acetaminophen + codeine (14.4%). At the time of the index, 1300 (20.6%) patients received some opioid. During the follow-up, a total of 1437 (22.8%) patients received a new opioid, of which 31.8% belonged to the group that received an initial opioid and 20.4% to the group that did not receive one, which was statistically significant (OR:1.81; 95%CI:1.58-2.07; p < 0.001). CONCLUSIONS: The frequent use of opioids in the management of migraines is potentially inappropriate and can lead to problems of tolerance, abuse and dependence. This combined with the low prescription of triptans, offers an opportunity for improvements in medical practice.

Functional Status of Patients over 65 Years Old Intervened on for a Hip Fracture One Year after the Operation.

OBJECTIVES: Evaluation of the functional status one year after a hip fracture surgery and the influence of sarcopenia and other clinical factors at the time of admission. METHOD: Prospective observational study with 135 patients over 65 years of age. Functional status of basic (modified Katz) and instrumental activities (Lawton and Brody) and walking ability (Functional Ambulation Classification, FAC) was measured on admission, at discharge, and telephonically one year later. The risk or positive screening of sarcopenia (SARC-F) and cognitive status (Pfeiffer), as well as clinical variables, were evaluated. RESULTS: 72% of patients are women; 36% have a risk of sarcopenia (Sarc-F ≥ 4), and 43% have moderate-severe cognitive impairment (Pfeiffer ≥ 5). Walking capacity at one year was closer to the values at admission more often in women than in men (0.2 ± 1.3 points vs. 0.9 ± 1.6; p = 0.001), as well as in patients without risk of sarcopenia versus sarcopenic patients (0.3 ± 1.2 points vs. 0.7 ± 1.7; p = 0.001), although their evolution did not show significant differences (p = 0.183). Instrumental activities after one year have not been recovered (1.7 ± 2.5 points; p = 0.032), and patients at risk of sarcopenia showed worse values (1.7 ± 1.9 points vs. 3.7 ± 2.7; p < 0.001) and worse evolution (p = 0.012). The evolution of basic activities varied according to the risk of sarcopenia (0.6 ± 1.4 points vs. 1.4 ± 2.1; p = 0.008). CONCLUSIONS: Functional status at one year is related to the functional status at admission, the positive screening of sarcopenia, sex, and cognitive impairment of the patient. Knowing at the time of admission an estimate of the functional status at one year will help to reinforce the individual treatment of patients with a worse prognosis.

Effectiveness of 2 interventions for independent oral feeding in preterms.

INTRODUCTION: Oral feeding of preterm newborns (PTNB) is hampered by their immaturity and intercurrent diseases, which can prolong their hospital stay. The objective of this study was to assess the effectiveness of a program that combines tactile, kinesthetic and oral stimulation (T + K + OS) compared to another intervention based on exclusively oral stimulation (OS), in the time necessary to achieve independent feeding and hospital discharge. PATIENTS AND METHODS: A clinical study of 2 randomized groups (OS vs. T + K + OS) was carried out on 42 PTNB with gestational age between 27-32 weeks and birth weight > 900 g. The stimulation programs were carried out in sessions of 15 min, for 10 days. RESULTS: The PTNBs in the T + K + OS group achieved independent oral feeding earlier, compared to the OS group (24.9 ± 10.1 vs. 34.1 ± 15.6 days, P = .02). An analysis of covariance was performed, which confirmed that the birth weight and gestational age covariates had significant effects on time to reach suction feeding (birth weight: F[1, 38] = 5.79; P = .021; gestational age: F[1, 38] = 14.12; P = .001) and that once its effect was controlled, the intervention continued to have a significant effect (F[1, 38] = 6.07; P = .018). The T + K + OS group, compared to the OS group, achieved an earlier hospital discharge (39 ± 15 vs. 45 ± 18 days), although the differences were not significant (P = .21). CONCLUSIONS: Combined therapies that include T + K + OS are more effective than OS alone, in order to achieve independent oral feeding in PTNBs.

The neural circuits of monogamous behavior.

The interest in studying the neural circuits related to mating behavior and mate choice in monogamous species lies in the parallels found between human social structure and sexual behavior and that of other mammals that exhibit social monogamy, potentially expanding our understanding of human neurobiology and its underlying mechanisms. Extensive research has suggested that social monogamy, as opposed to non-monogamy in mammals, is a consequence of the neural encoding of sociosensory information from the sexual partner with an increased reward value. Thus, the reinforced value of the mate outweighs the reward value of mating with any other potential sexual partners. This mechanism reinforces the social relationship of a breeding pair, commonly defined as a pair bond. In addition to accentuated prosocial behaviors toward the partner, other characteristic behaviors may appear, such as territorial and partner guarding, selective aggression toward unfamiliar conspecifics, and biparental care. Concomitantly, social buffering and distress upon partner separation are also observed. The following work intends to overview and compare known neural and functional circuits that are related to mating and sexual behavior in monogamous mammals. We will particularly discuss reports on Cricetid rodents of the Microtus and Peromyscus genus, and New World primates (NWP), such as the Callicebinae subfamily of the titi monkey and the marmoset (Callithrix spp.). In addition, we will mention the main factors that modulate the neural circuits related to social monogamy and how that modulation may reflect phenotypic differences, ultimately creating the widely observed diversity in social behavior.

Case report: Anterior open bite correction treatment by dental treatment and physical therapy through craniocervical mandibular and occlusal stabilization.

BACKGROUND: The opinion on whether a patient with an anterior open bite should be treated surgically or not is controversial. These patients generally suffer from associated discomfort due to their occlusal instability and musculoskeletal pain. CLINICAL PRESENTATION: A 60-year-old woman visited the clinic with dental mobility of her upper central incisors as her chief complaint. She had a severe anterior open bite, with a history of continuous grinding and multiple dental restorations in poor condition. Additionally, she suffered neck pain with movement restrictions. CONCLUSION: Dentists can evaluate and treat patients with an anterior open bite using this integrative model (physical therapy/dentistry) as a possible alternative as part of the treatment for anterior open bite patients.

[Effectiveness of 2 interventions for independent oral feeding in pre-terms]. / Eficacia de 2 intervenciones para la alimentación oral independiente en pretérminos.

INTRODUCTION: Oral feeding of pre-term newborns (PTNB) is hampered by their immaturity and intercurrent diseases, which can prolong their hospital stay. The objective of this study was to assess the effectiveness of a program that combines tactile, kinesthetic and oral stimulation (T+K+OS) compared to another intervention based on exclusively oral stimulation (OS), in the time necessary to achieve independent feeding and hospital discharge. PATIENTS AND METHODS: A clinical study of 2 randomized groups (OS vs. T+K+OS) was carried out on 42 PTNB with gestational age between 27-32 weeks and birth weight>900g. The stimulation programs were carried out in sessions of 15min, for 10 days. RESULTS: The PTNBs in the T+K+OS group achieved independent oral feeding earlier, compared to the OS group (24.9±10.1 vs. 34.1±15.6 days, P=.02). An analysis of covariance was performed, which confirmed that the birth weight and gestational age covariates had significant effects on time to reach suction feeding (birth weight: F[1, 38]=5.79; P=.021; gestational age: F[1, 38]=14.12; P=.001) and that once its effect was controlled, the intervention continued to have a significant effect (F[1, 38]=6.07; P=.018). The T+K+OS group, compared to the OS group, achieved an earlier hospital discharge (39±15 vs. 45±18 days), although the differences were not significant (P=.21). CONCLUSIONS: Combined therapies that include T+K+OS are more effective than OS alone, in order to achieve independent oral feeding in PTNBs.

Health-care guidelines and policies during the COVID-19 pandemic in Mexico: A case of health-inequalities.

BACKGROUND: Heterogeneous government responses have been reported in reaction to COVID-19. The aim of this study is to generate an exploratory review of healthcare policies published during COVID-19 by health-care institutions in Mexico. Analyzing policies within different health sub-systems becomes imperative in the Mexican case due to the longstanding fragmentation of the health-care system and health inequalities. DATA AND METHODS: Policies purposely included in the analysis were published by four public health institutions (IMSS, ISSSTE, SSA and PEMEX) during the COVID-19 epidemic in Mexico (from February 29th to June 15th, 2020) on official institutional websites. Researchers reviewed each document and classified them into seven policy categories set by the Rapid Research Evaluation and Appraisal Lab (RREAL): public health response, health-care delivery, human resources, health-system infrastructure and supplies, clinical response, health-care management, and epidemiological surveillance. RESULTS: Policy types varied by health institution. The largest number of policies were aimed at public health responses followed by health-care delivery and human resources. Policies were mainly published during the community transmission phase. CONCLUSIONS: The pandemic exposed underlying health-care system inequalities and a reactive rather than prepared response to the outbreak. Additionally, this study outlines potential policy gaps and delays in the response that could be avoided in the future.

Molecular epidemiology and genetic diversity of hepatitis B virus genotype E in an isolated Afro-Colombian community.

Hepatitis B virus (HBV) infection is a significant public health concern with 350 million chronic carriers worldwide. Eight HBV genotypes (A-H) have been described so far. Genotype E (HBV/E) is widely distributed in West Africa and has rarely been found in other continents, except for a few cases in individuals with an African background. In this study, we characterized HBV genotypes in Quibdó, Colombia, by partial S/P gene sequencing, and found, for the first time, HBV/E circulating in nine Afro-Colombian patients who had no recent contact with Africa. The presence of HBV/E in this community as a monophyletic group suggests that it was a result of a recent introduction by some Afro-descendent contact or, alternatively, that the virus came with slaves brought to Colombia. By using sequences with sampling dates, we estimated the substitution rate to be about 3.2 x 10(-4) substitutions per site per year, which resulted in a time to the most recent common ancestor (TMRCA) of 29 years. In parallel, we also estimated the TMRCA for HBV/E by using two previously estimated substitution rates (7.7 x 10(-4) and 1.5 x 10(-5) substitutions per site per year). The TMRCA was around 35 years under the higher rate and 1500 years under the slower rate. In sum, this work reports for the first time the presence of an exclusively African HBV genotype circulating in South America. We also discuss the time of the entry of this virus into America based on different substitution rates estimated for HBV.

Molecular characterization, distribution, and dynamics of hepatitis C virus genotypes in blood donors in Colombia.

Hepatitis C virus (HCV) is a frequent cause of acute and chronic hepatitis and a leading cause for cirrhosis of the liver and hepatocellular carcinoma. HCV is classified in six major genotypes and more than 70 subtypes. In Colombian blood banks, serum samples were tested for anti-HCV antibodies using a third-generation ELISA. The aim of this study was to characterize the viral sequences in plasma of 184 volunteer blood donors who attended the "Banco Nacional de Sangre de la Cruz Roja Colombiana," Bogotá, Colombia. Three different HCV genomic regions were amplified by nested PCR. The first of these was a segment of 180 bp of the 5'UTR region to confirm the previous diagnosis by ELISA. From those that were positive to the 5'UTR region, two further segments were amplified for genotyping and subtyping by phylogenetic analysis: a segment of 380 bp from the NS5B region; and a segment of 391 bp from the E1 region. The distribution of HCV subtypes was: 1b (82.8%), 1a (5.7%), 2a (5.7%), 2b (2.8%), and 3a (2.8%). By applying Bayesian Markov chain Monte Carlo simulation, it was estimated that HCV-1b was introduced into Bogotá around 1950. Also, this subtype spread at an exponential rate between about 1970 to about 1990, after which transmission of HCV was reduced by anti-HCV testing of this population. Among Colombian blood donors, HCV genotype 1b is the most frequent genotype, especially in large urban conglomerates such as Bogotá, as is the case in other South American countries.

Genomic analysis of two ORF2 segments of new porcine astrovirus isolates and their close relationship with human astroviruses.

Porcine astrovirus (PAstV) has been poorly studied and has been associated mainly with gastroenteritis. Computational analysis has revealed the close relationship of PAstV with astroviruses of humans (HAstV) and cats (FAstV). In this study, 105 and 171 stool specimens were collected from piglets and children under 5 years of age, respectively, in different Colombian regions during a 1-year period. The stool samples were examined for astroviruses by ELISA and RT-PCR; 23.8% and 4% were found to be positive for PAstV and HAstV, respectively. Additional sequence analysis with partial sequences obtained from ORF2 identified at least 2 probable groups of PAstVs and possible recombination events between porcine and human astroviruses. This study provides preliminary evidence of the high presence of PAstVs in pigs and proposes the existence of multiple PAstV types that are evolutionarily closely related to HAstVs.

Prevalence and molecular epidemiology of bovine leukemia virus in Colombian cattle.

Bovine leukemia virus (BLV) is one of the five agents considered most significant for cattle. It is important to determine the prevalence and molecular epidemiology of BLV throughout the country in order to gain a more thorough understanding of the current situation of BLV and to reveal the possibility of masked genotypes that the primers used by OIE are unable to identify. Blood samples were collected at random from 289 cows distributed in 75 farms across the country. PCR amplification of env, gag and tax gene segments was performed. The obtained amplicons were sequenced and then subjected to phylogenetic analyses. A total of 62% of the cows present at 92% of the farms were BLV-positive for gag fragment. Genotype 1 was exclusively detected by env gene segment when analyzed using previously reported primers. However, tax gene analysis revealed circulation of genotype 6 variants, which were also detected based on env gene analysis with newly designed primers. These results indicate that current genotyping approaches based on partial env sequencing may bias BLV genetic variability approaches and underestimate the diversity of the detected BLV genotypes. This report is one of the first molecular and epidemiological studies of BLV conducted in Colombia, which contributes to the global epidemiology of the virus; it also highlights the substantial impact of BLV on the country's livestock and thus is a useful resource for farmers and government entities.

[Phylogenetic analysis indicates human origin of rotavirus and hepatitis A virus strains found in the drinking water of western Colombia]. / Análisis filogenético de las cepas de rotavirus y virus de la hepatitis A encontradas en agua de consumo en el municipio de Quibdó, Chocó.

INTRODUCTION: Quibdó, the capital of Chocó Province, is one of the poorest cities in Colombia. Enteric viruses such as rotavirus and hepatitis A virus was found to occur commonly in city drinking water and indicated poor water quality and high risk of becoming infected. The source of these viruses was unknown, but humans and cattle were suspect sources. OBJECTIVE: City water was assessed to determine source and persistence of diarrhea and hepatitis among the human populations in the environs of Quibdó. MATERIAL AND METHODS: Four thousand liters of water were collected, filtered by tangential ultrafiltration and centrifuged in Centriprep Ultracel YM-50 tubes. Sixty samples of untreated and 20 of treated water were probed by RT-PCR. RESULTS: Six samples were positive for rotavirus and 2 for hepatitis A virus in both, treated and non treated water. DNA sequence analysis identified the presence of human G2 rotavirus and human hepatitis A virus. CONCLUSION: The evidence indicated a high level of contamination with pathogenic viruses in consumable water sources in Quibdó, Colombia.

Polyglandular syndrome type 2 in a Mexican family and its association with human leukocyte antigen.

The evaluation of first-degree family members is very important to detect additional cases of polyglandular autoimmune syndrome type 2. The genetic evaluation of human leukocyte antigen (HLA) may be useful in the study of this syndrome. This study is the first report of an HLA study of this syndrome in a Mexican family.

In silico and in vitro analysis of boAP3d1 protein interaction with bovine leukaemia virus gp51.

The envelope glycoprotein 51 (gp51) is essential for bovine leukaemia virus (BLV) entry to bovine B-lymphocytes. Although the bovine adaptor protein 3 complex subunit delta-1 (boAP3D1) has been proposed as the potential receptor, the specific ligand-receptor interaction has not yet been completely defined and boAP3D1 receptor and gp51 3D structures have not been determined. This study was thus aimed at a functional annotation of boAP3D1 cellular adaptor protein and BLV gp51 and, proposing a reliable model for gp51-AP3D1 interaction using bioinformatics tools. The boAP3D1 receptor interaction patterns were calculated based on models of boAP3D1 receptor and gp51 complexes' 3D structures, which were constructed using homology techniques and data-driven docking strategy. The results showed that the participation of 6 key amino acids (aa) on gp51 (Asn170, Trp127, His115, Ala97, Ser98 and Glu128) and 4 aa on AP3D1 (Lys925, Asp807, Asp695 and Arg800) was highly probable in the interaction between gp51 and BLVR domains. Three gp51 recombinant peptides were expressed and purified to validate these results: the complete domain (rgp51), the N-terminal portion (rNgp51) and the C-terminal fragment (rCgp51); and binding assays to Madin-Darby bovine kidney (MDBK) cells were then carried out with each recombinant. It was found that rNgp51 preferentially bound to MDBK cells, suggesting this domain's functional role during invasion. The rNgp51-MDBK cell interaction was sensitive to trypsin (98% reduction) and chymotrypsin treatment (80% reduction). These results highlighted that the N-terminal portion of gp51 interacted in vitro with the AP3D1 receptor and provides a plausible in silico interaction model.