Mucosal and systemic neutralizing antibodies to norovirus induced in infant mice orally inoculated with recombinant rotaviruses

Rotaviruses (RVs) preferentially replicate in the small intestine and frequently cause severe diarrheal disease, and the following enteric infection generally induces variable levels of protective systemic and mucosal immune responses in humans and other animals. Rhesus rotavirus (RRV) is a simian RV that was previously used as a human RV vaccine and has been extensively studied in mice. Although RRV replicates poorly in the suckling mouse intestine, infection induces a robust and protective antibody response. The recent availability of plasmid only-based RV reverse genetics systems has enabled the generation of recombinant RVs expressing foreign proteins. However, recombinant RVs have not yet been experimentally tested as potential vaccine vectors to immunize against other gastrointestinal pathogens in vivo. This is a newly available opportunity because several live-attenuated RV vaccines are already widely administered to infants and young children worldwide. To explore the feasibility of using RV as a dual vaccine vector, we rescued replication-competent recombinant RRVs harboring bicistronic gene segment 7 that encodes the native RV nonstructural protein 3 (NSP3) protein and a human norovirus (HuNoV) VP1 protein or P domain from the predominant genotype GII.4. The rescued viruses expressed HuNoV VP1 or P protein in infected cells in vitro and elicited systemic and local antibody responses to HuNoV and RRV following oral infection of suckling mice. Serum IgG and fecal IgA from infected suckling mice bound to and neutralized both RRV and HuNoV. These findings have encouraging practical implications for the design of RV-based next-generation multivalent enteric vaccines to target HuNoV and other human enteric pathogens.

Mucosal immunity plays a critical role in protecting against many pathogens in the respiratory and intestinal tracts. Live virus infections generally trigger more robust and effective mucosal immune response than oral administration of inactivated viruses or target protein antigens because they are self-amplifying and can more effectively elicit cellular as well as humoral immunity (1–4). Several studies have attempted to utilize recombinant viruses as vaccine vectors to induce an immune response against enteric pathogens (5–8); however, the most advanced of such enteric vaccine vectors are still in early stages of clinical development.Rotaviruses (RVs), the leading cause of acute gastroenteritis in infants, are a promising candidate for enteric vaccine vectors for several reasons. A) RV preferentially replicates in the small intestine, distinguishing it from several other enteric viruses that can also infect systemically or the colon. B) RV infection is acute, and the virus does not integrate into the host genome. C) RV is highly immunogenic and induces both systemic and mucosal immune responses in infected animals and humans (9, 10). D) Several live-attenuated human RV vaccines have been shown to be both safe and effective to use in very young children [e.g., RotaTeq (Merck) and Rotarix (GlaxoSmithKline)]. Other effective live-attenuated RV vaccines [Rotasiil, Rotavac, Lanzhou lamb rotavirus vaccine (LLR), and Rotavin-M1] are also licensed for use globally or primarily in their country of origin (11). E) Following substantial public health efforts, RV vaccines are now widely available in many low- and middle-income countries, as well as the more developed countries, and hence the administration of RV-based vaccines that included other heterologous antigens could potentially be piggybacked onto current RV immunization programs used globally. F) The RV double-stranded RNA (dsRNA) genome is segmented in nature, permitting easy genetic manipulation. G) With the insertion of heterologous antigens, RV replication can become attenuated in vitro (12, 13).Since a plasmid-based reverse genetics system was established in 2017, several studies have reported the generation of recombinant RVs that express fluorescent and bioluminescent reporter proteins (GFP, RFP, luciferase, etc.) and exogenous nucleotide sequences [e.g., endoribonuclease Csy4 target sequence and sequences encoding the receptor binding domain of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein] in vitro (12–22). To facilitate the assessment and development of RVs as potential enteric vaccine vectors, the capacity of recombinant RVs to induce an enteric immune response against other gastrointestinal (GI) pathogens needs to be evaluated in well-characterized preclinical small animal models. Rhesus rotavirus (RRV) is a prototype laboratory strain of simian RV that efficiently replicates in vitro (23, 24). Although RRV does not replicate well in a murine model (25–27), it does induce both systemic and mucosal immune responses in infected mice (28). In addition, RRV itself and RRV-based RV vaccine candidates have previously been shown to be a highly immunogenic and protective in several human vaccine trials and were, for a time, licensed for use in children in the United States (29, 30).Human norovirus (HuNoV) is a major cause of acute gastroenteritis in both young children and adults. Although B cells and human intestinal organoids support HuNoV replication (31, 32), there is not yet a widely available robust cell culture system for efficient HuNoV cultivation, which has impeded both the assessment of HuNoV immunity and vaccine development. The HuNoV virion consists of major capsid protein VP1 and minor capsid protein VP2 surrounding a positive-sense RNA genome (33–35). Exogenously expressed VP1 can form virus-like particles (VLPs) that are structurally and antigenically similar to HuNoV virions (36–38), and the parenteral administration of such VLPs provides some level of protective immunity to HuNoV in adults (39–41). Moreover, expression of the protruding or P domain of VP1 that bears the major antigenic sites of HuNoV can yield subunit “P particles” that can also induce immune responses (42, 43). Here, we demonstrate the induction of both systemic and mucosal antibody responses against HuNoV in suckling mice using recombinant RRVs expressing HuNoV VP1 or P domain. Our data suggest that recombinant RVs represent a potentially effective small-intestine–targeted vaccination platform to express exogenous genes in the human intestine and to protect people from other enteric pathogens such as HuNoV as well as RV.     

Intermittent theta‐burst stimulation rescues dopamine‐dependent corticostriatal synaptic plasticity and motor behavior in experimental parkinsonism: Possible role of glial activity

Background : Recent studies support the therapeutic utility of repetitive transcranial magnetic stimulation in Parkinson's disease (PD), whose progression is correlated with loss of corticostriatal long‐term potentiation and long‐term depression. Glial cell activation is also a feature of PD that is gaining increasing attention in the field because astrocytes play a role in chronic neuroinflammatory responses but are also able to manage dopamine (DA) levels. Methods : Intermittent theta‐burst stimulation protocol was applied to study the effect of therapeutic neuromodulation on striatal DA levels measured by means of in vivo microdialysis in 6‐hydroxydopamine‐hemilesioned rats. Effects on corticostriatal synaptic plasticity were studied through in vitro intracellular and whole‐cell patch clamp recordings while stepping test and CatWalk were used to test motor behavior. Immunohistochemical analyses were performed to analyze morphological changes in neurons and glial cells. Results : Acute theta‐burst stimulation induced an increase in striatal DA levels in hemiparkinsonian rats, 80 minutes post‐treatment, correlated with full recovery of plasticity and amelioration of motor performances. With the same timing, immediate early gene activation was restricted to striatal spiny neurons. Intense astrocytic and microglial responses were also significantly reduced 80 minutes following theta‐burst stimulation. Conclusion : Taken together, these results provide a first glimpse on physiological adaptations that occur in the parkinsonian striatum following intermittent theta‐burst stimulation and may help to disclose the real potential of this technique in treating PD and preventing DA replacement therapy‐associated disturbances. © 2017 International Parkinson and Movement Disorder Society     

Multivariate analyses of patient financial systems and oral health‐related quality of life

Johansson V, Axtelius B, Söderfeldt B, Sampogna F, Paulander J, Sondell K. Multivariate analyses of patient financial systems and oral health‐related quality of life. Community Dent Oral Epidemiol 2010; 38: 436–444. © 2010 John Wiley & Sons A/S Abstract – Objectives: Since 1999, the public dental health service (PDHS) in the county of Värmland, Sweden, has two co‐existing patient financial systems, i.e. ways for the patient to pay for dental care services. Alongside the traditional system of fee‐for‐service payment, i.e. paying afterwards for provided services, a new system of contract care is offered. In this system, dental care is covered by a contractual agreement, for which the patient pays an annual fee and receives care covered by the contract without additional costs. The aim of this article was to study whether patient financial system was associated with oral health‐related quality of life (OHRQoL). Methods: A questionnaire was answered by 1324 randomly selected patients, 52% from contract care and 48% from fee‐for‐service. The questionnaire contained questions about how much one was prepared to pay for dental care, how much one paid for dental care the previous year, OHIP‐14 (measured OHRQoL), dental anxiety, humanism of caregiver, SF‐36 (measured general health), multidimensional health locus of control, sense of coherence (SOC), self‐esteem and demographics. Data on patient financial system, gender and age were obtained from the sampling frame. The material was analysed with a hierarchical block method of multiple regression analysis. Results: When controlling for all other variables, patient financial system was one of the strongest associations with OHRQoL: patients in fee‐for‐service had worse OHRQoL than those in contract care. OHRQoL was also associated with general health, SOC and to some extent also with psychological and economic factors. Of the social variables, only being foreign born was significant: it was associated with worse OHRQoL. Conclusions: Patient financial system was associated with OHRQoL when controlling for confounding factors: patients in contract care had better OHRQoL than those in fee‐for‐service care.     

Effectiveness of Propranolol in the Treatment of Infantile Hemangioma Beyond the Proliferation Phase

During the last 5 years, many studies have shown the efficacy of propranolol as first‐line treatment for infantile hemangiomas (IHs), but not much has been written about the role of propranolol beyond the proliferation phase of IH (>1 year). Our aim was to assess propranolol efficacy and safety in the treatment of patients older than 1 year. A retrospective study of patients older than 1 year diagnosed with IH and treated in our vascular anomalies clinic between 2009 and 2013 was performed. Eighteen patients older than 1 year with a diagnosis of IH (15 girls, 3 boys) were identified. The mean age at the time of initiation of treatment was 25.7 months (range 13–72 mos). Single lesions were observed in 13 patients and multiple lesions in 5. Fifteen patients had focal lesions and three had segmental. The median duration of treatment with oral propranolol was 11.8 months (range 2–33 mos). Complete response was observed in 72.2% of the patients and partial response in 27.8%. Recurrence was observed in three patients 4.7 months after completion of therapy (range 0.3–8 mos). These patients required further therapy with propranolol for 6 more months. Bradycardia was documented in two patients and night terrors in one patient, which led to discontinuation of treatment. In our experience, propranolol may be useful in the treatment of IHs beyond the proliferation phase (>1 year old), but more studies are needed to support this observation.     

Isolated symptomatic cutaneous disease in hypereosinophilic syndrome

A 41-year-old Phillipino man presented with a 3-year history of a relapsing and remitting generalized chronic pruritic erythematous papular and plaque-like eruption. Investigations showed a persistently elevated eosinophil count. His disease was limited to cutaneous involvement with an absence of demonstrable internal organ involvement, despite extensive investigations and multidisciplinary review. Other causes of eosinophilia were excluded. A diagnosis of idiopathic hypereosinophilic syndrome was made. Our patient's presentation raises a number of issues related to hypereosinophilic syndrome. In particular, relating to managing hypereosinophilic syndrome and the challenge of minimizing therapy side-effects. Our case highlights the considerable morbidity of untreated isolated cutaneous disease, for which he was hospitalized with suicidal ideations. In a minority of reports, skin involvement is the only manifestation of hypereosinophilic syndrome.     

Shorter Telomere Length in Peripheral Blood Cells Associated With Migraine in Women

(Headache 2010;50:965‐972) Objective.— To evaluate relative telomere length of female migraine patients. Background.— Migraine is a debilitating disorder affecting 6‐28% of the population. Studies on the mechanisms of migraine have demonstrated genetic causes but the pathophysiology and subcellular effects of the disease remain poorly understood. Shortened telomere length is associated with age‐related or chronic diseases, and induced stresses. Migraine attacks may impart significant stress on cellular function, thus this study investigates a correlation between shortening of telomeres and migraine. Methods.— Relative telomere length was measured using a previously described quantitative polymerase chain reaction method. A regression analysis was performed to assess differences in mean relative telomere length between migraine patients and healthy controls. Results.— The leukocyte telomeres of a cohort of 142 Caucasian female migraine subjects aged 18‐77 years and 143 matched 17‐77‐year‐old healthy control Caucasian women were examined. A significantly shorter relative telomere length was observed in the migraine group compared with the control group after adjusting for age and body mass index (P = .001). In addition, age of onset was observed to associate with the loss of relative telomere length, especially at early age of onset (<17 years old). No association was observed between relative telomere length and the severity and frequency of migraine attacks and the duration of migraine. Conclusion.— Telomeres are shorter in migraine patients and there is more variation in telomere length in migraine patients.