Barcoding Brazilian mammals to monitor biological diversity and threats: Trends, perspectives, and knowledge gaps.

DNA barcoding and environmental DNA (eDNA) represent significant advances for biomonitoring the world's biodiversity and its threats. However, these methods are highly dependent on the presence of species sequences on molecular databases. Brazil is one of the world's largest and most biologically diverse countries. However, many knowledge gaps still exist for describing, identifying, and monitoring of mammalian biodiversity using molecular methods. We aimed to unravel the patterns of the presence of Brazilian mammal species on molecular databases to improve our understanding of how effectively it would be to monitor them using DNA barcoding and environmental DNA, and contribute to mammalian conservation. We foundt many gaps in molecular databases, with many taxa being poorly represented, particularly from Amazonia, the order Lagomorpha, and arboreal, gomivorous, near extinct, and illegally traded species. Moreover, our analyses revealed that species description year was the most important factor determining the probability of a species to being sequenced. Primates are the group with the highest number of species considered a priority for sequencing due to their high level of combined threats. We highlight where investments are needed to fill knowledge gaps and increase the representativity of species on molecular databases to enable a better monitoring ability of Brazilian mammals encompassing different traits using DNA barcoding and environmental DNA.

In vivo rAAV-mediated human TGF-ß overexpression reduces perifocal osteoarthritis and improves osteochondral repair in a large animal model at one year.

OBJECTIVE: Osteoarthritis (OA) is a serious consequence of focal osteochondral defects. Gene transfer of human transforming growth factor beta (hTGF-ß) with recombinant adeno-associated virus (rAAV) vectors offers a strategy to improve osteochondral repair. However, the long-term in vivo effects of such rAAV-mediated TGF-ß overexpression including its potential benefits on OA development remain unknown. METHOD: Focal osteochondral defects in minipig knees received rAAV-lacZ (control) or rAAV-hTGF-ß in vivo. After one year, osteochondral repair and perifocal OA were visualized using validated macroscopic scoring, ultra-high-field MRI at 9.4 T, and micro-CT. A quantitative estimation of the cellular densities and a validated semi-quantitative scoring of histological and immunohistological parameters completed the analysis of microarchitectural parameters. RESULTS: Direct rAAV-hTGF-ß application induced and maintained significantly improved defect filling and safranin O staining intensity and overall cartilage repair at one year in vivo. In addition, rAAV-hTGF-ß led to significantly higher chondrocyte densities within the cartilaginous repair tissue without affecting chondrocyte hypertrophy and minimized subarticular trabecular separation. Of note, rAAV-hTGF-ß significantly improved the adjacent cartilage structure and chondrocyte density and reduced overall perifocal OA development after one year in vivo. CONCLUSIONS: rAAV-hTGF-ß treatment improves long-term osteochondral repair and delays the progression of perifocal OA in a translational model. These findings have considerable potential for targeted molecular approaches to treat focal osteochondral defects.

[Endoprosthetic replacement following intercalary resection]. / Endoprothetische Rekonstruktion nach interkalarer Resektion.

BACKGROUND: Endoprosthetic replacement is a valuable treatment option following intercalary resection of bone tumours in the diaphysis. OBJECTIVES: To identify indication, operative technique, implants currently available, literature results and alternative procedures for the alloplastic reconstruction of segmental bone defects. MATERIALS AND METHODS: This review article summarizes the authors' own experiences and relevant clinical studies focussing on this topic. RESULTS: According to the literature, 10-year-survival rates of intercalary endoprostheses range between 64 and 80%. Yet, comparisons between different publications are difficult due to the limited number of cases, different implants, follow-up periods and the heterogeneous patient populations. Biological alternatives for reconstruction are autologous bone transplantation, distraction osteogenesis and bone transport, allogenic bone transplantation, and the induced membrane technique. Innovative tissue engineering approaches are still limited to preclinical testing. CONCLUSIONS: Short- to mid-term results for segmental endoprostheses following intercalary resections are satisfactory and may be regarded as superior to those of biological reconstructions due to the immediate full weight-bearing capability. However, they are mainly applied for elderly patients and in palliative situations because of potential long-term complications.

Bernese periacetabular osteotomy through a double approach : Simplification of a surgical technique.

OBJECTIVE: Reorientation of the acetabulum to normalize load transfer or avert femoroacetabular pincer impingement to prevent osteoarthritis of the hip. INDICATIONS: Persisting acetabular dysplasia after closure of growth plates or acetabular malrotation. CONTRAINDICATIONS: High dislocation of hip, secondary acetabulum, increased misalignment on functional X­ray, high-grade mobility restriction. Relative: degenerative changes, advanced age. SURGICAL TECHNIQUE: Bernese periacetabular osteotomy through two incisions; all bone cuts are carried out under direct vision. The osteotomies are equivalent to the classic Ganz method. In a slightly tilted forward lateral decubitus position, a posterior incision is applied for the ischium osteotomy and the caudal portion of the retroacetabular osteotomy. The pubis and ilium osteotomies are performed in a supine position through an anterior approach with subsequent reorientation and screw fixation. The rectus femoris is not dissected unless joint exposure is required. POSTOPERATIVE MANAGEMENT: Partial weight bearing with 20 kg for the first 6 weeks postoperatively, followed by stepwise transition to full loads after radiological control. RESULTS: In total, 34 patients (37 hips) were followed up for 20.4 ± 10.3 months. Tönnis osteoarthritis scale levels remained constant. The center-edge angle of Wiberg increased from 13.2 ± 7.5° to 26.5 ± 6.7°, the Tönnis angle (acetabular index) changed from 13.8 ± 6.5° to 3.4 ± 4.4°. At follow-up, the Merle d'Aubigné and Postel score was 16.5 ± 1.4; the modified Harris hip score 87.6 ± 13.9 and the International hip outcome tool (iHOT)-12 78.2 ± 20.3 points. The mean surgical time was 213 ± 29 min. Severe complications were not observed.

[Meniscal lesion. A pre-osteoarthritic condition of the knee joint]. / Die Meniskusläsion ­ ein präarthrotischer Zustand des Kniegelenks.

BACKGROUND: A close relationship between meniscal damage and articular cartilage exist. Likewise, (partial) meniscectomy may lead to the development of osteoarthritis (OA). OBJECTIVES: With a special emphasis on therapeutic consequences for orthopaedic surgeons, the structural and functional relationship between meniscal tears/extrusion and cartilage loss, and/or the effect of meniscectomy or meniscal repair on the development of OA, are emphasized. MATERIALS AND METHODS: A selective literature review with implementation of own research findings. RESULTS: The close topographical and functional interplay between the menisci and the tibiofemoral cartilage is the basis for the clinically important relationship between meniscal damage and cartilage degeneration. In particular, due to its close connection to tibiofemoral OA, a degenerative meniscal lesion represents a pre-osteoarthritic condition. Meniscus extrusion is also often associated with tibiofemoral OA. Even large cartilage defects can cause meniscus lesions. Partial meniscectomy is strongly associated with the incidence and risk of progression of OA. Clinical results are particularly problematic after partial resection of the lateral meniscus. Although the use of arthroscopic partial resection for degenerative meniscal lesions has been controversially discussed, no long-term studies are available. A large number of studies emphasize the medium-term value of meniscus reconstruction compared to partial meniscus resection. Combined meniscus and cartilage damage are complex cases, and the value of a simultaneous therapy remains unclear. CONCLUSIONS: Preserving the meniscus is the first step towards cartilage repair. Randomized and controlled studies will provide better information on the long-term outcomes of meniscal resection and repair with regard to OA development.

Macroscopic cartilage repair scoring of defect fill, integration and total points correlate with corresponding items in histological scoring systems - a study in adult sheep.

OBJECTIVE: To correlate osteochondral repair assessed by validated macroscopic scoring systems with established semiquantitative histological analyses in an ovine model and to test the hypothesis that important macroscopic individual categories correlate with their corresponding histological counterparts. METHODS: In the weight-bearing portion of medial femoral condyles (n = 38) of 19 female adult Merino sheep (age 2-4 years; weight 70 ± 20 kg) full-thickness chondral defects were created (size 4 × 8 mm; International Cartilage Repair Society (ICRS) grade 3C) and treated with Pridie drilling. After sacrifice, 1520 blinded macroscopic observations from three observers at 2-3 time points including five different macroscopic scoring systems demonstrating all grades of cartilage repair where correlated with corresponding categories from 418 blinded histological sections. RESULTS: Categories "defect fill" and "total points" of different macroscopic scoring systems correlated well with their histological counterparts from the Wakitani and Sellers scores (all P ≤ 0.001). "Integration" was assessed in both histological scoring systems and in the macroscopic ICRS, Oswestry and Jung scores. Here, a significant relationship always existed (0.020 ≤ P ≤ 0.049), except for Wakitani and Oswestry (P = 0.054). No relationship was observed for the "surface" between histology and macroscopy (all P > 0.05). CONCLUSIONS: Major individual morphological categories "defect fill" and "integration", and "total points" of macroscopic scoring systems correlate with their corresponding categories in elementary and complex histological scoring systems. Thus, macroscopy allows to precisely predict key histological aspects of articular cartilage repair, underlining the specific value of macroscopic scoring for examining cartilage repair.

2D and 3D MOCART scoring systems assessed by 9.4 T high-field MRI correlate with elementary and complex histological scoring systems in a translational model of osteochondral repair.

OBJECTIVE: To compare the 2D and 3D MOCART system obtained with 9.4 T high-field magnetic resonance imaging (MRI) for the ex vivo analysis of osteochondral repair in a translational model and to correlate the data with semiquantitative histological analysis. METHODS: Osteochondral samples representing all levels of repair (sheep medial femoral condyles; n = 38) were scanned in a 9.4 T high-field MRI. The 2D and adapted 3D MOCART systems were used for grading after point allocation to each category. Each score was correlated with corresponding reconstructions between both MOCART systems. Data were next correlated with corresponding categories of an elementary (Wakitani) and a complex (Sellers) histological scoring system as gold standards. RESULTS: Correlations between most 2D and 3D MOCART score categories were high, while mean total point values of 3D MOCART scores tended to be 15.8-16.1 points higher compared to the 2D MOCART scores based on a Bland-Altman analysis. "Defect fill" and "total points" of both MOCART scores correlated with corresponding categories of Wakitani and Sellers scores (all P ≤ 0.05). "Subchondral bone plate" also correlated between 3D MOCART and Sellers scores (P < 0.001). CONCLUSIONS: Most categories of the 2D and 3D MOCART systems correlate, while total scores were generally higher using the 3D MOCART system. Structural categories "total points" and "defect fill" can reliably be assessed by 9.4 T MRI evaluation using either system, "subchondral bone plate" using the 3D MOCART score. High-field MRI is valuable to objectively evaluate osteochondral repair in translational settings.

Experimental scoring systems for macroscopic articular cartilage repair correlate with the MOCART score assessed by a high-field MRI at 9.4 T--comparative evaluation of five macroscopic scoring systems in a large animal cartilage defect model.

OBJECTIVE: To develop a new macroscopic scoring system which allows for an overall judgment of experimental articular cartilage repair and compare it with four existing scoring systems and high-field magnetic resonance imaging (MRI). METHODS: A new macroscopic scoring system was developed to assess the repair of cartilage defects. Cartilage repair was graded by three observers with different experience in cartilage research at 2-3 time points and compared with the protocol A of the international cartilage repair society (ICRS) cartilage repair assessment score, the Oswestry arthroscopy score, and macroscopic grading systems designed by Jung and O'Driscoll. Parameters were correlated with the two-dimensional (2D) magnetic resonance observation of cartilage repair tissue (MOCART) score based on a 9.4 T MRI as an external reference standard. RESULTS: All macroscopic scores exhibited high intra- and interobserver reliability and high internal correlation. The newly developed macroscopic scoring system had the highest intraobserver [0.866 ≤ intraclass correlation (ICC) ≤ 0.895] and the highest interobserver reliability (ICC = 0.905) for "total points". Here, Cronbach's alpha indicated good homogeneity and functioning of the items (mean = 0.782). "Total points" of the 2D MOCART score correlated with all macroscopic scores (all P < 0.0001). The newly developed macroscopic scoring system yielded the highest correlation for the MRI parameter "defect fill" (rho = 0.765; all P < 0.0001). CONCLUSIONS: "Total points" and "defect fill", two clinically relevant indicators of cartilage repair, can be reliably and directly assessed by macroscopic evaluation, using either system. These data support the use of macroscopic assessment to precisely judge cartilage repair in preclinical large animal models.

Correlates of youth smokeless tobacco use.

The objectives of this study are to identify knowledge and attitude variables that correlate with smokeless tobacco use and how youth users and non-users differ in their attitudes and knowledge about smokeless tobacco. A randomized cluster sample of 1834 total fifth, eighth, and 11th grade students in West Virginia public schools during the 1996-1997 school year were surveyed on smokeless tobacco knowledge, attitudes and practices. Data from 648 male non-users and 160 male monthly and daily users of smokeless tobacco were compared using chi 2 and two-tailed t-tests. Logistic regression analysis of survey variables revealed the following correlates of smokeless tobacco use: having a family member not living in the home who uses, having a friend who uses, playing football, having tried cigarettes, and having parents who would permit use at home. Important differences exist in knowledge and attitudes regarding smokeless tobacco between users and non-users in fifth, eighth, and 11th grade West Virginia public schools. Correlates of smokeless tobacco use are identified which can be used to target prevention programs.

A peer review feedback method of promoting compliance with preventive care guidelines in a resident ambulatory care clinic.

BACKGROUND: Simple distribution of clinical practice guidelines to physicians does not change practice behavior. A low-cost, continuous peer review feedback method was used to promote resident physicians' compliance with nine preventive care guidelines at the ambulatory care clinic at the Marshall University School of Medicine (Huntington, West Virginia). METHODS: Preventive care guidelines were distributed and a peer review feedback program was instituted in the resident physician primary care practice. The frequency of resident physician use of nine preventive care services was assessed and compared during three periods: preguideline (September 1, 1993, to March 1, 1994; 148 patients), guideline (September 1, 1994, to March 1, 1995; 148 patients), and one-year follow-up (September 1, 1995, to March 1, 1996; 150 patients). The patients in the three periods were similar in age, gender, and risk for influenza and pneumococcal infection. RESULTS: During the guideline period, resident physicians offered patients four preventive care services-tetanus toxoid immunization, clinical breast examination, Papanicolaou smear testing, and hemoccult testing significantly more often than during the preguideline period. All services were offered significantly more often during the one-year follow-up period compared with the preguideline period and as often as in the guideline period. CONCLUSION: A low-cost, continuous peer review feedback program significantly and durably improves resident physician compliance with clinical practice guidelines on preventive care services. However, the effectiveness of the poor review feedback method may not generalize to private practice or other settings. Research on other methods to promote compliance with clinical practice guidelines and to influence physician behavior in general should continue.

Ribotype analysis of Pseudomonas pseudomallei isolates.

No epidemiological typing system to differentiate among Pseudomonas pseudomallei isolates has been available. Ribotype analysis was developed and used to examine 74 clinical and 10 environmental isolates of P. pseudomallei from Thailand. Six P. pseudomallei ribotypes were identified from restriction fragment polymorphisms of EcoRI chromosomal digests. The predominant ribotype, A, was found in 59 of the isolates examined. By using patterns from hybridizations with SalI, HindIII, and PstI restriction digests, isolates of ribotype A were subdivided into a further five subtypes, giving a total of 10 differentiable P. pseudomallei types. In 23 of 34 melioidosis patients studied, multiple P. pseudomallei isolates were present. In all but one of these patients, a single ribotype of the organism was present. Isolation of two different ribotypes of P. pseudomallei from one patient, one each in sputum and urine, suggests that superinfection may have occurred. The ribotype was shown to be conserved during the course of antibiotic treatments in seven patients studied, although the antibiotic sensitivity patterns in the isolates from these patients varied. The prevalence of subtype A1 in clinical and environmental specimens suggests that this strain may be predominant in this geographical location. These results demonstrate the usefulness of the ribotyping method for epidemiological studies of P. pseudomallei.