Assessment of the Composition and Biologic Activity of Platelet Rich Plasma and its Relationship to Clinical Outcomes in Patients with Knee Osteoarthritis

Recent studies suggest positive clinical outcomes associated with platelet-rich-plasma (PRP) administration to treat knee osteoarthritis (OA). However, the results remain inconclusive in part because of the high variability in PRP preparations and the limited information regarding the relevant biologically active components of PRP. We hypothesize that the variability in clinical response is driven by the heterogeneous composition of PRP. In this study we evaluated the composition and biological activity of PRP and further correlated our findings to clinical outcomes in patients receiving intra-articular injections for knee OA. After IRB approval and patient consent, we enrolled 51 patients (mean age: 57.9 ± 10.1; mean BMI: 26.0 ± 4.1) with mild-moderate knee OA (Kellgren Lawrence grades 1-3), eligible for intra-articular PRP injection. We obtained MRI at baseline and outcome measures (KOOS JR and PNS) at baseline, 6 weeks, 6 months, and 12 months after PRP injection. Patients were categorized as “good” and “poor” responders based on the outcome measures, corrected using published Minimally Clinically Important Difference (MCID) values. Aliquots of PRP and whole blood from the same patients were used to evaluate composition (CBC with differential and multiplex ELISA) and biologic activity, using a co-culture system of macrophages and fibroblast incubated with TNFa with and without PRP (10% v:v) for 24 hours. Total RNA from cells was used for RNAseq, Nanostring, and RTqPCR analysis. On average, we collected 4.07 ± 01.05 mL of PRP, and 3.24 ± 0.85 mL of PRP were injected intra-articularly. PRP preparations yielded mean fold-changes of 1.60 ± 0.37 platelets and 0.19 ± 0.08v leukocytes, relative to whole-blood from the same patients (set as 1). On average, all patients that reached the 6-month time-point (N = 32) reported improved outcomes at 6-weeks and 6-months after PRP administration (KOOS and PNS p\u3c0.05 vs. baseline). After MCID corrections, we identified “good” (N=17, positive response using both measures) and “poor” responders (N=15, poor response in one or both measures). RNAseq analyses showed PRP-dependent changes in the TNFa-induced modulation of a number of genes, including CXCL7 and CCL5. NanoString and RTqPCR analyses confirmed the RNAseq results. Comparisons of PRP from good and poor responders identified changes in the composition and biologic activity between these groups. This pilot study integrated clinical data with genomic approaches to evaluate variability in the composition and activity of PRP, and how this may influence outcomes in patients with knee OA. We uncovered subsets of genes differentially modulated by co-treatment of PRP with TNFa, in agreement with the concept that the reduced knee OA pain in patients treated with PRP is driven by the ability of PRP to modulate inflammation. Furthermore, we identified changes in the composition and biologic activity of PRP between “good” and “poor” responders

Evaluation of Rotator Cuff Repair with Concomitant Biceps Tenodesis

Introduction: Surgical rotator cuff repair (RCR) has proven to be an effective treatment for rotator cuff tears. Commonly, rotator cuff tears are associated with concomitant biceps pathology, which are often treated by biceps tenodesis (BT). We hypothesize that patient outcomes will be similar in those that have undergone RCR with concomitant BT and isolated RCR. Methods: This is a retrospective cohort study comparing patients who underwent arthroscopic RCR with arthroscopic or open BT to patients who underwent isolated RCR at a multisurgeon orthopaedic practice during the time period of November 2016 to December 2016. The outcome for comparison is the American Shoulder and Elbow Surgeons score (ASES). Patients with postoperative scores of at least 6 months after surgery were included. The data was collected from the Rothman Institute registry and OBERD. It was analyzed via independent t-test. Results: A total of 53 patients (37 = M; 16 = F) were in the isolated RCR group and 34 patients (27 = M; 7 = F) were in the RCR with BT group. The average age in the isolated RCR group was 58.6 years vs. 58.9 years in the RCR with BT group. There was no statistical difference between postoperative ASES scores (83.69 and 79.43, P = .40) and difference in preoperative and postoperative ASES scores (34.26 and 35.30, P = .85) in the isolated RCR and RCR with BT groups, respectively. Conclusion: There was no significant difference in postoperative ASES scores as well as difference in preoperative and postoperative ASES scores in patients undergoing isolated RCR and RCR with BT. This supports the hypothesis that patients undergoing RCR with BT will have similar outcomes to those undergoing isolated RCR

Genetic analysis of Thailand hantavirus in Bandicota indica trapped in Thailand

Sixty one tissue samples from several rodent species trapped in five provinces of Thailand were examined for the presence of hantaviral markers by enzyme-immunoassay and immunoblotting. Four samples, all from the great bandicoot rat Bandicota indica, were confirmed positive for the hantaviral N-antigen. Two of them were trapped in Nakhon Pathom province, the other two in Nakhon Ratchasima province, approximately 250 km from the other trapping site. When analysed by RT-nested PCR, all four rodents were found positive for the hantaviral S- and M-segment nucleotide sequences. Genetic analysis revealed that the four newly described wild-type strains belong to Thailand hantavirus. On the phylogenetic trees they formed a well-supported cluster within the group of Murinae-associated hantaviruses and shared a recent common ancestor with Seoul virus

Hantavirus Infection in the Republic of Georgia

We describe a laboratory-confirmed case of hantavirus infection in the Republic of Georgia. Limited information is available about hantavirus infections in the Caucasus, although the infection has been reported throughout Europe and Russia. Increasing awareness and active disease surveillance contribute to our improved understanding of the geographic range of this pathogen

Genetic evidence of Dobrava virus in Apodemus agrarius in Hungary.

Using nested polymerase chain reaction, we sequenced Dobrava virus (DOB) from the rodent Apodemus agrarius in Hungary. The samples we isolated group with DOB samples previously isolated from A. flavicollis. This grouping may indicate host switching

Juquitiba-like Hantavirus from 2 Nonrelated Rodent Species, Uruguay

Serologic and genetic analyses indicate that a Juquitiba-like hantavirus circulates in Maldonado, Uruguay. This virus is carried by 2 rodent species, Oligoryzomys nigripes and Oxymycterus nasutus. The same hantavirus in 2 nonrelated species can be explained by a spillover infection or a host-switching event

Molecular characterization of highly pathogenic H5N1 avian influenza viruses isolated in Sweden in 2006

<p>Abstract</p> <p>Background</p> <p>The analysis of the nonstructural (NS) gene of the highly pathogenic (HP) H5N1 avian influenza viruses (AIV) isolated in Sweden early 2006 indicated the co-circulation of two sub-lineages of these viruses at that time. In order to complete the information on their genetic features and relation to other HP H5N1 AIVs the seven additional genes of twelve Swedish isolates were amplified in full length, sequenced, and characterized.</p> <p>Results</p> <p>The presence of two sub-lineages of HP H5N1 AIVs in Sweden in 2006 was further confirmed by the phylogenetic analysis of approximately the 95% of the genome of twelve isolates that were selected on the base of differences in geographic location, timing and animal species of origin. Ten of the analyzed viruses belonged to sub-clade 2.2.2. and grouped together with German and Danish isolates, while two 2.2.1. sub-clade viruses formed a cluster with isolates of Egyptian, Italian, Slovenian, and Nigerian origin. The revealed amino acid differences between the two sub-groups of Swedish viruses affected the predicted antigenicity of the surface glycoproteins, haemagglutinin and neuraminidase, rather than the nucleoprotein, polymerase basic protein 2, and polymerase acidic protein, the main targets of the cellular immune responses. The distinctive characteristics between members of the two subgroups were identified and described.</p> <p>Conclusion</p> <p>The comprehensive genetic characterization of HP H5N1 AIVs isolated in Sweden during the spring of 2006 is reported. Our data support previous findings on the coincidental spread of multiple sub-lineage H5N1 HPAIVs via migrating aquatic birds to large distance from their origin. The detection of 2.2.1. sub-clade viruses in Sweden adds further data regarding their spread in the North of Europe in 2006. The close genetic relationship of Swedish isolates sub-clade 2.2.2. to the contemporary German and Danish isolates supports the proposition of the introduction and spread of a single variant of 2.2.2. sub-clade H5N1 avian influenza viruses in the Baltic region. The presented findings underline the importance of whole genome analysis.</p

Preclinical proof of concept of a tetravalent lentiviral T-cell vaccine against dengue viruses

Dengue virus (DENV) is responsible for approximately 100 million cases of dengue fever annually, including severe forms such as hemorrhagic dengue and dengue shock syndrome. Despite intensive vaccine research and development spanning several decades, a universally accepted and approved vaccine against dengue fever has not yet been developed. The major challenge associated with the development of such a vaccine is that it should induce simultaneous and equal protection against the four DENV serotypes, because past infection with one serotype may greatly increase the severity of secondary infection with a distinct serotype, a phenomenon known as antibody-dependent enhancement (ADE). Using a lentiviral vector platform that is particularly suitable for the induction of cellular immune responses, we designed a tetravalent T-cell vaccine candidate against DENV (“LV-DEN”). This vaccine candidate has a strong CD8+ T-cell immunogenicity against the targeted non-structural DENV proteins, without inducing antibody response against surface antigens. Evaluation of its protective potential in the preclinical flavivirus infection model, i.e., mice knockout for the receptor to the type I IFN, demonstrated its significant protective effect against four distinct DENV serotypes, based on reduced weight loss, viremia, and viral loads in peripheral organs of the challenged mice. These results provide proof of concept for the use of lentiviral vectors for the development of efficient polyvalent T-cell vaccine candidates against all DENV serotypes