Molecular Investigation of Canine Parvovirus-2 (CPV-2) Outbreak in Nevis Island: Analysis of the Nearly Complete Genomes of CPV-2 Strains from the Caribbean Region

To date, there is a dearth of information on canine parvovirus-2 (CPV-2) from the Caribbean region. During August–October 2020, the veterinary clinic on the Caribbean island of Nevis reported 64 household dogs with CPV-2-like clinical signs (hemorrhagic/non-hemorrhagic diarrhea and vomiting), of which 27 animals died. Rectal swabs/fecal samples were obtained from 43 dogs. A total of 39 of the 43 dogs tested positive for CPV-2 antigen and/or DNA, while 4 samples, negative for CPV-2 antigen, were not available for PCR. Among the 21 untested dogs, 15 had CPV-2 positive littermates. Analysis of the complete VP2 sequences of 32 strains identified new CPV-2a (CPV-2a with Ser297Ala in VP2) as the predominant CPV-2 on Nevis Island. Two nonsynonymous mutations, one rare (Asp373Asn) and the other uncommon (Ala262Thr), were observed in a few VP2 sequences. It was intriguing that new CPV-2a was associated with an outbreak of gastroenteritis on Nevis while found at low frequencies in sporadic cases of diarrhea on the neighboring island of St. Kitts. The nearly complete CPV-2 genomes (4 CPV-2 strains from St. Kitts and Nevis (SKN)) were reported for the first time from the Caribbean region. Eleven substitutions were found among the SKN genomes, which included nine synonymous substitutions, five of which have been rarely reported, and the two nonsynonymous substitutions. Phylogenetically, the SKN CPV-2 sequences formed a distinct cluster, with CPV-2b/USA/1998 strains constituting the nearest cluster. Our findings suggested that new CPV-2a is endemic in the region, with the potential to cause severe outbreaks, warranting further studies across the Caribbean Islands. Analysis of the SKN CPV-2 genomes corroborated the hypothesis that recurrent parallel evolution and reversion might play important roles in the evolution of CPV-2.     

Human Protoparvovirus DNA and IgG in Children and Adults with and without Respiratory or Gastrointestinal Infections

Three human protoparvoviruses, bufavirus (BuV), tusavirus (TuV) and cutavirus (CuV), have recently been discovered in diarrheal stool. BuV has been associated with diarrhea and CuV with cutaneous T-cell lymphoma, but there are hardly any data for TuV or CuV in stool or respiratory samples. Hence, using qPCR and IgG enzyme immunoassays, we analyzed 1072 stool, 316 respiratory and 445 serum or plasma samples from 1098 patients with and without gastroenteritis (GE) or respiratory-tract infections (RTI) from Finland, Latvia and Malawi. The overall CuV-DNA prevalences in stool samples ranged between 0–6.1% among our six patient cohorts. In Finland, CuV DNA was significantly more prevalent in GE patients above rather than below 60 years of age (5.1% vs 0.2%). CuV DNA was more prevalent in stools among Latvian and Malawian children compared with Finnish children. In 10/11 CuV DNA-positive adults and 4/6 CuV DNA-positive children with GE, no known causal pathogens were detected. Interestingly, for the first time, CuV DNA was observed in two nasopharyngeal aspirates from children with RTI and the rare TuV in diarrheal stools of two adults. Our results provide new insights on the occurrence of human protoparvoviruses in GE and RTI in different countries.     

Forced-eruption time for palatally impacted canines treated with and without ostectomy-decortication technique

Objectives:To compare forced-eruption times for palatally impacted canines treated with and without the ostectomy-decortication technique and to assess the influence of palatally impacted canine pretreatment position and angle on forced-eruption time.Materials and Methods:The sample was composed of 118 patient-subjects with 151 palatally impacted canines treated with the ostectomy-decortication technique (n = 72) and without (n = 79). The orthopantomogram radiographs (OPGs) were analyzed for palatally impacted canine angle and horizontal and vertical position. Recovery time was measured from the start of forced eruption until the canine was within ±1 mm of final dental arch position.Results:The time of forced canine eruption with ostectomy-decortication technique was significantly shorter than without (6.6 vs 21.0 months). Pretreatment canine position significantly increased forced-eruption time in the ostectomy-decortication group but not in the control sample.Conclusions:Forced-eruption time of palatally impacted canines using the ostectomy-decortication technique was 3.2 times more rapid than without. Forced-eruption time increased significantly as a function of pretreatment palatally impacted canine position severity in the ostectomy-decortication group but not in the control.     

Pharmacokinetic aspects and in vitro–in vivo correlation potential for lipid-based formulations

Lipid-based formulations have been an attractive choice among novel drug delivery systems for enhancing the solubility and bioavailability of poorly soluble drugs due to their ability to keep the drug in solubilized state in the gastrointestinal tract. These formulations offer multiple advantages such as reduction in food effect and inter-individual variability, ease of preparation, and the possibility of manufacturing using common excipients available in the market. Despite these advantages, very few products are available in the present market, perhaps due to limited knowledge in the in vitro tests (for prediction of in vivo fate) and lack of understanding of the mechanisms behind pharmacokinetic and biopharmaceutical aspects of lipid formulations after oral administration. The current review aims to provide a detailed understanding of the in vivo processing steps involved after oral administration of lipid formulations, their pharmacokinetic aspects and in vitro in vivo correlation (IVIVC) perspectives. Various pharmacokinetic and biopharmaceutical aspects such as formulation dispersion and lipid digestion, bioavailability enhancement mechanisms, impact of excipients on efflux transporters, and lymphatic transport are discussed with examples. In addition, various IVIVC approaches towards predicting in vivo data from in vitro dispersion/precipitation, in vitro lipolysis and ex vivo permeation studies are also discussed in detail with help of case studies.KEY WORDS: Pharmacokinetics, Lipolysis, IVIVC, Efflux transporters, Lymphatic delivery, Food effectAbbreviations: ADME, absorption/distribution/metabolism/elimination; AUC, area under the curve; BCS, biopharmaceutics classification system; BDDCS, biopharmaceutics drug disposition classification system; CACO, human epithelial colorectal adenocarcinoma cells; C_max, maximum plasma concentration; CMC, critical micellar concentration; CYP, cytochrome; DDS, drug delivery systems; FaSSGF, fasted-state simulated gastric fluid; FaSSIF, fasted-state simulated intestinal fluid; FeSSIF, fed-state simulated intestinal fluid; GIT, gastrointestinal tract; IVIVC, in vitro in vivo correlation; LCT, long chain triglyceride; LFCS, lipid formulation classification system; log P, n-octanol/water partition coefficient; MCT, medium chain triglyceride; MDCK, Madin–Darby canine kidney cells; NCE, new chemical entity; P-app, apparent permeability; P-gp, permeability glycoprotein; SCT, short chain triglyceride; SEDDS, self-emulsifying drug delivery system; SIF, simulated intestinal fluid; SMEDDS, self-microemulsifying drug delivery system; SNEDDS, self-nanoemulsifying drug delivery system; Vit E, vitamin E     

Cone-beam computerized tomography evaluation of canine retraction using micro implant and conventional anchorage

ObjectiveVarious anchorage techniques have been designed for canine retraction. The aim was to measure and compare the rate of canine retraction with conventional method and micro implant using CBCT.Materials and methodsSample size comprising of 17 subjects were scheduled for extraction of all first premolars. After leveling and aligning, titanium micro implants were placed between the roots of the second premolar and the first molars on right side, in both the arches. Pre and post retraction CBCT scans were taken. Retraction was done using sliding mechanics using stainless steel arch wire.ResultsThe maxillary right canine (micro implant) retracted by 6.75 mm and tipped distally by 9.51° at a rate of 1.05 mm/month while the mandibular right canine retracted by 4.83 mm and tipped distally by 7.88° at a rate of 1.13 mm/month. On the left side (conventional) with molar as a source of anchorage, maxillary canine retracted by 6.03 mm and tipped distally by 6.51° at a rate of 1.46 mm/month while the mandibular canine retracted by 5.03 mm and tipped distally by 4.34° at a rate of 1.15 mm/month.ConclusionImplants can serve as a source of anchorage.     

Treatment Strategies for Missing Maxillary Central Incisor—An Orthodontist's Perspective

The loss of maxillary central incisors at an early age has psychological, esthetic, and functional implications. Multiple treatment options are available for replacing missing central incisors. The management demands a multidisciplinary approach involving the orthodontist, prosthodontist, and periodontist. Treatment planning requires consideration of a variety of clinical and nonclinical factors. This clinical report attempts to demonstrate different strategies for the management of unilaterally and bilaterally missing central incisors.     

Evaluation of Molecular Test for the Discrimination of “Naked” DNA from Infectious Parvovirus B19 Particles in Serum and Bone Marrow Samples

Low levels of parvovirus B19 (B19V) DNA can be detected in the circulation and in different tissue of immunocompetent individuals for months or years, which has been linked to inflammatory diseases such as cardiomyopathy, rheumatoid arthritis, hepatitis, and vasculitis. However, the detection of B19V DNA does not necessarily imply that infectious virions are present. This study aimed to evaluate the method based on the Benzonase^® treatment for differentiation between the infectious virions from “naked” DNA in serum and bone marrow (BM) samples to be useful for the B19V routine diagnosis. In addition, we estimated the period of viremia and DNAemia in the sera and bone marrow of nonhuman primates experimentally infected with B19V. Serum samples from ten patients and from four cynomolgus monkeys experimentally infected with B19V followed up for 60 days were used. Most of the human serum samples became negative after pretreatment; however, only decreased viral DNA loads were observed in four patients, indicating that these samples still contained the infectious virus. Reduced B19V DNA levels were observed in animals since 7th dpi. At approximately 45th dpi, B19V DNA levels were below 10⁵ IU/mL after Benzonase^® pretreatment, which was not a consequence of active B19V replication. The test based on Benzonase^® pretreatment enabled the discrimination of “naked DNA” from B19V DNA encapsidated in virions. Therefore, this test can be used to clarify the role of B19V as an etiological agent associated with atypical clinical manifestations.     

尖牙的生理性与病理性迁徙的临床分析

目的：探讨阻生尖牙与迁徙尖牙的联系、生理性迁徙或病理性迁徙与阻生尖牙的关系及迁徙尖牙的治疗方法。方法：对历年的X线片资料进行分析并复习有关文献。结果：阻生尖牙迁徙者有12例（上颌2例、下颌10例）,其中生理性迁徙5例,病理性迁徙7例。结论：部分尖牙阻生可能为迁徙发生的早期阶段,尖牙迁徙包括生理性迁徙和病理性迁徙,尖牙生理性迁徙和病理性迁徙在发病原因、临床表现和治疗方法上都明显不同。