Immunogenic fusion proteins induce neutralizing SARS-CoV-2 antibodies in the serum and milk of sheep

Antigen-specific polyclonal immunoglobulins derived from the serum, colostrum, or milk of immunized ruminant animals have potential as scalable therapeutics for the control of viral diseases such as COVID-19. Enhancing the efficacy of vaccine antigens to induce robust and specific antibody responses remains central to developing highly effective formulations. The direct fusion of immunoglobulin (IgG) Fc domains or other immune-stimulating proteins to antigens has shown promise in several mammalian species but has not yet been tested and optimized in commercially-relevant ruminant species. Here we show that the immunization of sheep with fusions of the receptor binding domain (RBD) of SARS-CoV-2 to ovine IgG2a Fc domains promotes significantly higher levels of antigen-specific antibodies compared to native RBD or full-length spike antigens. This antibody population was shown to contain elevated levels of neutralizing antibodies that suppress binding between the RBD and soluble hACE2 receptors in vitro. The parallel evaluation of a second immune-stimulating fusion candidate, Granulocyte-macrophage colony-stimulating factor (GM-CSF), induced high neutralizing responses in select animals but narrowly missed achieving significance at the group level. Furthermore, we demonstrate that the antibodies induced by these fusion antigens are transferred from maternal serum into colostrum/milk. These antibodies also demonstrate cross-neutralizing activity against diverse SARS-CoV-2 variants including delta and omicron. Our findings highlight a new pathway for recombinant antigen design in ruminant animals with applications in immune milk production and animal health.

Adipose-Derived Stem Cells, Obesity, and Inflammation: A Systematic Review and Implications for Osteoarthritis Treatment.

ABSTRACT: Adipose is a known source of mesenchymal stem cells that can be used to treat musculoskeletal disorders, such as osteoarthritis. Because obesity often coexists with osteoarthritis, excess adiposity may be a useful source of mesenchymal stem cells. However, obesity is associated with systemic inflammation, which may influence the quality of adipose-derived stem cells. We performed a systematic review of the literature examining adipose-derived stem cell behavior, cytokine, and growth factor profiles from obese and nonobese patients. Two independent reviewers applied the inclusion/exclusion criteria and independently extracted data including mesenchymal stem cell count/viability/behavior, growth factor, and/or cytokine expression. Twenty-two articles met criteria for inclusion. Samples from obese patients had increased mesenchymal stem cell content (n = 6), but decreased proliferative ability (n = 3), and increased expression of interleukin 1 (n = 3), interleukin 6 (n = 3), and tumor necrosis factor α (n = 6). There was also greater macrophage content (n = 4). Weight loss normalized cellular function. In vitro behavior and quality of adipose-derived stem cell are significantly different between obese and nonobese patients. Samples from obese patients had greater adipose-derived stem cell content, lower proliferative ability, increased senescence, and increased proinflammatory cytokine expression. Differences in cellular function should be considered when using adipose to treat musculoskeletal pathology in obese and nonobese patients.

Treatment decisions after interdisciplinary evaluation for nonarthritic hip pain: A randomized controlled trial.

BACKGROUND: Physical therapy and hip arthroscopy are two viable treatment options for patients with nonarthritic hip pain (NAHP); however, patients may experience considerable decisional conflict when making a treatment decision. Interdisciplinary evaluation with a physical therapist and surgeon may better inform the decision-making process and reduce decisional conflict. OBJECTIVE: To identify the extent to which an interdisciplinary evaluation between a surgeon, physical therapist, and patient influences treatment plans and decisional conflict of persons with NAHP. DESIGN: Randomized controlled trial. SETTING: Hip preservation clinic. PARTICIPANTS: Adults with primary NAHP. INTERVENTIONS: Participants were randomized to receive a standard (surgeon) or interdisciplinary (surgeon+physical therapist) evaluation. Surgeon evaluations included patient interview, strength and range-of-motion examination, palpation, gross motor observation, and special testing. Interdisciplinary evaluations started with the surgeon evaluation, then a physical therapist evaluated movement impairments during sitting, sit-to-stand, standing, single-leg stance, single-leg squat, and walking. All evaluations concluded with treatment planning with the respective provider(s). OUTCOME MEASURES: Treatment plan and decisional conflict were collected pre- and postevaluation. Inclusion of physical therapy in participants' postevaluation treatment plans and postevaluation decisional conflict were compared between groups using chi-square tests and Mann-Whitney U tests, respectively. RESULTS: Seventy-eight participants (39 in each group) met all eligibility criteria and were included in all analyses. Sixty-six percent of participants who received an interdisciplinary evaluation included physical therapy in their postevaluation treatment plan, compared to 48% of participants who received a standard evaluation (p = .10). Participants who received an interdisciplinary evaluation reported 6.3 points lower decisional conflict regarding their postevaluation plan (100-point scale; p = .04). The interdisciplinary and standard groups reduced decisional conflict on average 24.8 ± 18.9 and 23.6 ± 14.6 points, respectively. CONCLUSIONS: Adding a physical therapist to a surgical clinic increased interest in physical therapy treatment, but this increase was not statistically significant. The interdisciplinary group displayed lower postevaluation decisional conflict; however, both groups displayed similar reductions in decisional conflict from pre- to postevaluation. This study also demonstrated the feasibility of an interdisciplinary evaluation in a hip preservation clinic.

Predictive profiling of SARS-CoV-2 variants by deep mutational learning

The continual evolution of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and the emergence of variants that show resistance to vaccines and neutralizing antibodies (1-4) threaten to prolong the coronavirus disease 2019 (COVID-19) pandemic (5). Selection and emergence of SARS-CoV-2 variants are driven in part by mutations within the viral spike protein and in particular the ACE2 receptor-binding domain (RBD), a primary target site for neutralizing antibodies. Here, we develop deep mutational learning (DML), a machine learning-guided protein engineering technology, which is used to interrogate a massive sequence space of combinatorial mutations, representing billions of RBD variants, by accurately predicting their impact on ACE2 binding and antibody escape. A highly diverse landscape of possible SARS-CoV-2 variants is identified that could emerge from a multitude of evolutionary trajectories. DML may be used for predictive profiling on current and prospective variants, including highly mutated variants such as omicron (B.1.1.529), thus supporting decision making for public heath as well as guiding the development of therapeutic antibody treatments and vaccines for COVID-19.

High Intensity Interval Exercise Increases Platelet and Transforming Growth Factor-ß Yield in Platelet-Rich Plasma.

BACKGROUND: Platelet-rich plasma (PRP) is an emerging orthobiologic treatment for musculoskeletal conditions like osteoarthritis. Two studies have demonstrated the influence of longer duration exercise on PRP composition, but no study has ever explored the impact of high intensity interval exercise (HIIE) on PRP content. OBJECTIVE: To quantify cellular and growth factor content changes in PRP after 4 minutes of HIIE. DESIGN: Controlled laboratory pilot study. SETTING: Academic sports medicine center. PARTICIPANTS: Ten healthy volunteers (5 male, 5 female). INTERVENTION: Volunteers had PRP prepared from 15 mL of whole blood using a single spin, plasma-based system (autologous conditioned plasma [ACP]) immediately before and after 4 minutes of HIIE on a stationary exercise bike (Tabata protocol). MAIN OUTCOME MEASURE: The PRP was sent for complete blood counts and enzyme-linked immunosorbent assay (ELISA) to quantify transforming growth factor (TGF)-ß, platelet-derived growth factor (PDGF), insulin-like growth factor (IGF)-1, and vascular endothelial growth factor (VEGF). RESULTS: Mean platelet count in PRP increased from 367.4 ± 57.5 k/µL to 497.7 ± 93.3 k/µL after 4 minutes of HIIE (P < .001). TGF-ß also increased from 8237.2 ± 7676.5 pg/mL to 21 535.7 ± 4062.6 pg/mL postexercise (P = .004). The other cellular components (leukocytes, red blood cells, and mean platelet volume) and growth factors (PDGF, IGF-1, and VEGF) were not significantly changed. CONCLUSIONS: A short 4-minute bout of HIIE significantly increased the total platelet count and TGF-ß concentration in PRP.

Inguinal Hernia in Athletes: Role of Dynamic Ultrasound.

BACKGROUND: Inguinal hernia is a commonly encountered cause of pain in athletes. Because of the anatomic complexity, lack of standard imaging, and the dynamic condition, there is no unified opinion explaining its underlying pathology. HYPOTHESIS: Athletes with persistent groin pain would have a high prevalence of inguinal hernia with dynamic ultrasound, and herniorrhaphy would successfully return athletes to activity. STUDY DESIGN: Case-control study. LEVEL OF EVIDENCE: Level 3. METHODS: Forty-seven amateur and professional athletes with sports-related groin pain who underwent ultrasound were selected based on history and examination. Patients with prior groin surgery or hip pathology were excluded. Clinical and surgical documentation were correlated with imaging. The study group was compared with 41 age-matched asymptomatic athletes. RESULTS: Ultrasound was positive for hernia with movement of bowel, bladder, or omental tissue anterior to the inferior epigastric vessels during Valsalva maneuver. The 47-patient symptomatic study group included 41 patients with direct inguinal hernias, 1 with indirect inguinal hernia, and 5 with negative ultrasound. Of 42 patients with hernia, 39 significantly improved with herniorrhaphy, 2 failed to improve after surgery and were diagnosed with adductor longus tears, and 1 improved with physical therapy. Five patients with negative ultrasound underwent magnetic resonance imaging and were diagnosed with hip labral tear or osteitis pubis. The 41-patient asymptomatic control group included 3 patients with direct inguinal hernias, 2 with indirect inguinal hernias, and 3 with femoral hernias. CONCLUSION: Inguinal hernias are a major component of groin pain in athletes. Prevalence of direct inguinal hernia in symptomatic athletes was greater than that for controls ( P < 0.001). Surgery was successful in returning these athletes to sport: 39 of 42 (93%) athletes with groin pain and inguinal hernia became asymptomatic. CLINICAL RELEVANCE: Persistent groin pain in the athlete may relate to inguinal hernia, which can be diagnosed with dynamic ultrasound imaging. Herniorrhaphy is successful at returning athletes to sports activity.

Unrecognized pediatric partial Achilles tendon injury followed by traumatic completion: a case report and literature review.

Achilles tendon ruptures are a relatively common athletic injury but are exceedingly rare in the pediatric population. We describe the case of a 10-year-old ice hockey player who experienced an Achilles tendon injury from a laceration to the posterior leg from a skate blade that led to a partial tendon laceration. This tendon injury was initially unrecognized despite an emergency department evaluation. The patient continued to complain of weakness and paresthesia after the skin laceration had healed. A traumatic dorsiflexion injury while running several weeks later led to a traumatic complete tendon rupture. The clinical, operative, and physical therapy records were reviewed to complete the history, treatment, and rehabilitation progress. The initial laceration injury had occurred 6 weeks before presentation, and the traumatic dorsiflexion injury had occurred 2 days before referral to an acute orthopedics clinic. Open repair was performed several days after the traumatic completion of the laceration, and the patient was immobilized in a cast for 5 weeks. The patient had weaned off crutches by 10 weeks postoperatively and had returned to some activities and light skating at 5.5 months. A full return to running and ice hockey had been achieved by 8 months postoperatively. The optimal repair for this injury has not been well established in published studies. We have concluded that laceration injuries have the potential to mask tendon injuries and that prolonged symptoms after a laceration should suggest occult pathologic features. Open tendon repair is a viable treatment option in the pediatric patient with Achilles tendon ruptures. A return to activities within a reasonable period can be expected with robust physical therapy.