Adenoviral Vector-Based Vaccines and Gene Therapies: Current Status and Future Prospects

Adenoviruses are one of the most genetically diverse DNA viruses and cause non-life-threatening infections in the ocular, respiratory, or gastrointestinal epithelium of a diverse range of hosts. Adenoviruses are excellent vectors for delivering genes or vaccine antigens to the target host tissues and are being tested in several vaccine and gene therapy studies. Adenovirus-based vectors offer several advantages over other viral vectors such as broad range of tissue tropism, well-characterized genome, ease of genetic manipulation including acceptance of large transgene DNA insertions, inherent adjuvant properties, ability to induce robust transgene-specific T cell and antibody responses, non-replicative nature in host, and ease of production at large scale. However, several studies have highlighted major drawbacks to using adenovirus as vaccine and gene therapy vectors. These include pre-existing immunity in humans, inflammatory responses, sequestering of the vector to liver and spleen, and immunodominance of the vector genes over transgenes. In the same vein, recently discovered protein sequence homology and heterologous immunity between adenoviruses and hepatitis C virus have significant implications in the use of adenoviral vectors for vaccine development, especially for hepatitis C virus. This chapter focuses on the current scope and challenges in using adenoviral vector-based vaccines and gene therapies

CLN6 disease caused by the same mutation originating in Pakistan has varying pathology

AbstractThe neuronal ceroid lipofuscinoses (NCLs), the most common neurodegenerative diseases in children, are characterised by storage of autofluorescent material that has a characteristic ultrastructure. We report two families with variant late infantile NCL, both originating from Pakistan. Probands from both families were homozygous for the same mutation (c.316dupC) but had variable pathology to that currently thought to be typical for CLN6 disease, late infantile variant. The observed pathology of one proband resembled condensed fingerprints, previously described in late infantile CLN7 and CLN8 diseases, and pathology from the second proband was thought to be absent even after repeated skin biopsy, but observed after review. This mutation is the most common NCL mutation in families originating from Pakistan and could be prioritised for testing. Finally, this report contains the first prenatal diagnosis for late infantile CLN6 disease, initially made on the basis of EM and now confirmed by mutation analysis

Abolishing spontaneous epileptiform activity in human brain tissue through AMPA receptor inhibition

Objective: The amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) is increasingly recognized as a therapeutic target in drug-refractory pediatric epilepsy. Perampanel (PER) is a non-competitive AMPAR antagonist, and pre-clinical studies have shown the AMPAR-mediated anticonvulsant effects of decanoic acid (DEC), a major medium-chain fatty acid provided in the medium-chain triglyceride ketogenic diet. Methods: Using brain tissue resected from children with intractable epilepsy, we recorded the effects of PER and DEC in vitro. Results: We found resected pediatric epilepsy tissue exhibits spontaneous epileptic activity in vitro, and showed that DEC and PER inhibit this epileptiform activity in local field potential recordings as well as excitatory synaptic transmission. Interpretation: This study confirms AMPAR antagonists inhibit epileptiform discharges in brain tissue resected in a wide range of pediatric epilepsies

N-methyl-D-aspartate receptor antibody-mediated neurological disease:results of a UK-based surveillance study in children

OBJECTIVE: N-methyl-D-aspartate receptor antibody (NMDAR-Ab) encephalitis is a well-recognised clinico-immunological syndrome that presents with neuropsychiatric symptoms cognitive decline, movement disorder and seizures. This study reports the clinical features, management and neurological outcomes of paediatric NMDAR-Ab-mediated neurological disease in the UK. DESIGN: A prospective surveillance study. Children with NMDAR-Ab-mediated neurological diseases were voluntarily reported to the British Neurological Surveillance Unit (BPNSU) from November 2010 to December 2011. Initial and follow-up questionnaires were sent out to physicians. RESULTS: Thirty-one children fulfilled the criteria for the study. Eight presented during the study period giving an incidence of 0.85 per million children per year (95% CI 0.64 to 1.06); 23 cases were historical. Behavioural change and neuropsychiatric features were present in 90% of patients, and seizures and movement disorders both in 67%. Typical NMDAR-Ab encephalitis was reported in 24 children and partial phenotype without encephalopathy in seven, including predominantly psychiatric (four) and movement disorder (three). All patients received steroids, 22 (71%) received intravenous immunoglobulin, 9 (29%) received plasma exchange,and 10 (32%) received second-line immunotherapy. Of the 23 patients who were diagnosed early, 18 (78%) made a full recovery compared with only 1 of 8 (13%) of the late diagnosed patients (p=0.002, Fisher's exact test). Seven patients relapsed, with four needing additional second-line immunotherapy. CONCLUSIONS: Paediatric NMDAR-Ab-mediated neurological disease appears to be similar to adult NMDAR-Ab encephalitis, but some presented with a partial phenotype. Early treatment was associated with a quick and often full recovery

Phenotypic and genetic spectrum of epilepsy with myoclonic atonic seizures

Objective: We aimed to describe the extent of neurodevelopmental impairments andidentify the genetic etiologies in a large cohort of patients with epilepsy with myoclonicatonic seizures (MAE).Methods: We deeply phenotyped MAE patients for epilepsy features, intellectualdisability, autism spectrum disorder, and attention-deficit/hyperactivity disorderusing standardized neuropsychological instruments. We performed exome analysis(whole exome sequencing) filtered on epilepsy and neuropsychiatric gene sets toidentify genetic etiologies.Results: We analyzed 101 patients with MAE (70% male). The median age of seizureonset was 34 months (range = 6-72 months). The main seizure types were myoclonicatonic or atonic in 100%, generalized tonic-clonic in 72%, myoclonic in 69%, absencein 60%, and tonic seizures in 19% of patients. We observed intellectual disability in62% of patients, with extremely low adaptive behavioral scores in 69%. In addition,24% exhibited symptoms of autism and 37% exhibited attention-deficit/hyperactivitysymptoms. We discovered pathogenic variants in 12 (14%) of 85 patients, includingfive previously published patients. These were pathogenic genetic variants inSYNGAP1 (n = 3), KIAA2022 (n = 2), and SLC6A1 (n = 2), as well as KCNA2,SCN2A, STX1B, KCNB1, and MECP2 (n = 1 each). We also identified three newcandidate genes, ASH1L, CHD4, and SMARCA2 in one patient each.Significance: MAE is associated with significant neurodevelopmental impairment.MAE is genetically heterogeneous, and we identified a pathogenic genetic etiologyin 14% of this cohort by exome analysis. These findings suggest that MAE is a manifestationof several etiologies rather than a discrete syndromic entity

Classic ketogenic diet versus further antiseizure medicine in infants with drug-resistant epilepsy (KIWE): a UK, multicentre, open-label, randomised clinical trial

BACKGROUND: Many infancy-onset epilepsies have poor prognosis for seizure control and neurodevelopmental outcome. Ketogenic diets can improve seizures in children older than 2 years and adults who are unresponsive to antiseizure medicines. We aimed to establish the efficacy of a classic ketogenic diet at reducing seizure frequency compared with further antiseizure medicine in infants with drug-resistant epilepsy. METHODS: In this phase 4, open-label, multicentre, randomised clinical trial, infants aged 1-24 months with drug-resistant epilepsy (defined as four or more seizures per week and two or more previous antiseizure medications) were recruited from 19 hospitals in the UK. Following a 1-week or 2-week observation period, participants were randomly assigned using a computer-generated schedule, without stratification, to either a classic ketogenic diet or a further antiseizure medication for 8 weeks. Treatment allocation was masked from research nurses involved in patient care, but not from participants. The primary outcome was the median number of seizures per day, recorded during weeks 6-8. All analyses were by modified intention to treat, which included all participants with available data. Participants were followed for up to 12 months. All serious adverse events were recorded. The trial is registered with the European Union Drug Regulating Authorities Clinical Trials Database (2013-002195-40). The trial was terminated early before all participants had reached 12 months of follow-up because of slow recruitment and end of funding. FINDINGS: Between Jan 1, 2015, and Sept 30, 2021, 155 infants were assessed for eligibility, of whom 136 met inclusion criteria and were randomly assigned; 75 (55%) were male and 61 (45%) were female. 78 infants were assigned to a ketogenic diet and 58 to antiseizure medication, of whom 61 and 47, respectively, had available data and were included in the modifified intention-to-treat analysis at week 8. The median number of seizures per day during weeks 6-8, accounting for baseline rate and randomised group, was similar between the ketogenic diet group (5 [IQR 1-16]) and antiseizure medication group (3 [IQR 2-11]; IRR 1·33, 95% CI 0·84-2·11). A similar number of infants with at least one serious adverse event was reported in both groups (40 [51%] of 78 participants in the ketogenic diet group and 26 [45%] of 58 participants in the antiseizure medication group). The most common serious adverse events were seizures in both groups. Three infants died during the trial, all of whom were randomly assigned a ketogenic diet: one child (who also had dystonic cerebral palsy) was found not breathing at home; one child died suddenly and unexpectedly at home; and one child went into cardiac arrest during routine surgery under anaesthetic. The deaths were judged unrelated to treatment by local principal investigators and confirmed by the data safety monitoring committee. INTERPRETATION: In this phase 4 trial, a ketogenic diet did not differ in efficacy and tolerability to a further antiseizure medication, and it appears to be safe to use in infants with drug-resistant epilepsy. A ketogenic diet could be a treatment option in infants whose seizures continue despite previously trying two antiseizure medications. FUNDING: National Institute for Health and Care Research

Utilization of mechanical power and associations with clinical outcomes in brain injured patients: a secondary analysis of the extubation strategies in neuro-intensive care unit patients and associations with outcome (ENIO) trial

Background: There is insufficient evidence to guide ventilatory targets in acute brain injury (ABI). Recent studies have shown associations between mechanical power (MP) and mortality in critical care populations. We aimed to describe MP in ventilated patients with ABI, and evaluate associations between MP and clinical outcomes. Methods: In this preplanned, secondary analysis of a prospective, multi-center, observational cohort study (ENIO, NCT03400904), we included adult patients with ABI (Glasgow Coma Scale ≤ 12 before intubation) who required mechanical ventilation (MV) ≥ 24 h. Using multivariable log binomial regressions, we separately assessed associations between MP on hospital day (HD)1, HD3, HD7 and clinical outcomes: hospital mortality, need for reintubation, tracheostomy placement, and development of acute respiratory distress syndrome (ARDS). Results: We included 1217 patients (mean age 51.2 years [SD 18.1], 66% male, mean body mass index [BMI] 26.3 [SD 5.18]) hospitalized at 62 intensive care units in 18 countries. Hospital mortality was 11% (n = 139), 44% (n = 536) were extubated by HD7 of which 20% (107/536) required reintubation, 28% (n = 340) underwent tracheostomy placement, and 9% (n = 114) developed ARDS. The median MP on HD1, HD3, and HD7 was 11.9 J/min [IQR 9.2-15.1], 13 J/min [IQR 10-17], and 14 J/min [IQR 11-20], respectively. MP was overall higher in patients with ARDS, especially those with higher ARDS severity. After controlling for same-day pressure of arterial oxygen/fraction of inspired oxygen (P/F ratio), BMI, and neurological severity, MP at HD1, HD3, and HD7 was independently associated with hospital mortality, reintubation and tracheostomy placement. The adjusted relative risk (aRR) was greater at higher MP, and strongest for: mortality on HD1 (compared to the HD1 median MP 11.9 J/min, aRR at 17 J/min was 1.22, 95% CI 1.14-1.30) and HD3 (1.38, 95% CI 1.23-1.53), reintubation on HD1 (1.64; 95% CI 1.57-1.72), and tracheostomy on HD7 (1.53; 95%CI 1.18-1.99). MP was associated with the development of moderate-severe ARDS on HD1 (2.07; 95% CI 1.56-2.78) and HD3 (1.76; 95% CI 1.41-2.22). Conclusions: Exposure to high MP during the first week of MV is associated with poor clinical outcomes in ABI, independent of P/F ratio and neurological severity. Potential benefits of optimizing ventilator settings to limit MP warrant further investigation

A comparative study of ropivacaine 0.5% versus ropivacaine 0.75% for spinal anesthesia in lower limb orthopedic surgery in ASA Grade – I/II adult patients: A prospective study

Aims and Objectives: The aim of the study was to compare the clinical efficacy and safety of isobaric ropivacaine 0.5% and 0.75% in spinal anesthesia under: (a) Onset and duration of sensory and motor block, (b) duration of analgesia, and (c) adverse effects. Methods: A total of 60 patients undergoing elective lower limb orthopedic surgery under spinal anesthesia were divided into two groups (I and II) of 30 each. Group I received 3ml of isobaric ropivacaine 0.5% Group II received 3 ml of isobaric ropivacaine 0.75%. The study parameters were recorded at baseline and then at specified intervals. Statistics: By professional statisticians using SPSS 18 version. Student t-test was used for continuous variables, and Chi-square test was used for discrete variables. Results: The onset of sensory blockage in Group I was 3.17 ± 1.29 min and 2.60 ± 1.19 min in Group II which was statistically not significant (P > 0.05). The onset of motor blockade in Group I was 3.90 ± 1.54 min and 3.10 ± 0.96 min in Group II which was statistically significant (P < 0.05). Median time to reach the highest level of analgesia was 12.4 ± 2.81 min in Group I, and 10.7 ± 2.56 min in Group II. The difference was statistically significant. Regression of sensory level to T10 dermatome in Group I was 99.64 ± 21.30 min and 139.66 ± 25.70 min in Group II which was statistically significant (P < 0.05). Duration of the motor blockade in Group I was 126 ± 14.53 min and 175 ± 30.60 min in Group II which was statistically significant (P < 0.05). The time of the first request of analgesics in Group I was 130 ± 16.24 min and 171.1 ± 32.77 min in Group II which was statistically significant (P < 0.05). There were no significant differences in the adverse effects of both drugs. Conclusions: Intrathecal isobaric ropivacaine 0.75% in comparison to isobaric ropivacaine 0.5%: (1) Produces quicker onset of motor block and prolonged duration of sensory and motor block. (2) Does not alter hemodynamic stability. (3) Has no difference in the onset of sensory block.&nbsp

Combined spinal epidural for labor analgesia comparison of two different doses of intrathecal bupivacaine 1.25 mg and fentanyl 25 µg with bupivacaine 2.5 mg and fentanyl 25 µg

Background and Objective: The responsibility of the anesthetist in obstetrics is very high. This study compares two different low doses of intrathecal bupivacaine 1.25 mg and 2.5 mg along with 25 µg fentanyl as the spinal component of combined spinal epidural (CSE) analgesia in the early part of labor, followed by epidural top-up. Methodology: Approval was obtained from the institutional review board and written informed consent was obtained from 60 healthy term primigravida or the second gravid parturients, with cephalic singleton pregnancy between 36 and 42 weeks, ASA Grade I/II patients. The study was conducted using low-dose intrathecal bupivacaine 1.25 mg and fentanyl 25 µg (GroupI) with bupivacaine 2.5 mg and fentanyl 25 µg (Group II) as the spinal component of CSE analgesia in the early part of labor. We compared the two with respect to their onset, duration of sensory and motor block, quality of analgesia during early part of labor and the side effects of the drugs. Results: The onset of analgesia was equally rapid with both groups within 5 min, lower incidence of motor block with Group I compared to Group II. Duration of analgesia was longer in Group II, associated with higher dermatome levels of sensory block with longer time for regression of the block. Coclusion: We found that bupivacaine 1.25 mg was as effective as bupivacaine 2.5 mg when added to fentanyl 25 µg for CSE.&nbsp

Clinicopathological study of cerebellar astrocytoma in children

Introduction: The first successful treatment of a pediatric brain tumor was in 1879 when Sir William Macewen successfully removed a meningioma from a 14-year-old girl. Brain tumors are the most common form of solid tumors and the leading cause of death from solid tumors in children (SEER program 1975–1999). Materials and Methods: Study area: This study was conducted at Bangur Institute of Neuroscience (BIN) and S.S.K.M Hospital. Study Population: Patients attending BIN OPD and admitting in BIN and S.S.K.M Hospital wards were selected. Inclusion Criteria: The following criteria were included in the study: (a) Patients with the diagnosis of cerebellar astrocytomas after magnetic resonance imaging investigation, (b) patients giving consent to be included in the study, and (c) patient willing to come for follow up. Study Period: The study period was 2 years (from September 1, 2010, to December 31, 2012). Sample Size: All diagnosed cases of cerebellar astrocytoma during the stated period. Exclusion Criteria: Patients not willing for the study were excluded from the study. Study Design: This was a non-randomized prospective clinical study. Pilocytic astrocytomas are the most common pediatric brain tumors in our population and are most commonly located in the cerebellum. Results: Most of the patients, 20 (90.9%), had neurological improvement on discharge. 1 patient (4.5%) died during the hospital course. The follow-up time period ranged from 3 months to 2 years, with a mean follow-up period of 1.5 years. Recurrence was observed in 5 patients (22.72%), but reoperation was done in 3 patients (13.63%). Of them, 1 patient (4.5%) received radiotherapy in spite of that recurrence was developed in 1.5 years. 2 patients (9.09%) have been kept under observation because these are asymptomatic. The solid consistency of tumors led to a poor prognosis, as it was associated with a greater number of ICU admissions, recurrence of tumors, and repeat surgeries (13.63%). The follow-up time period ranged from 3 months to 2 years, with a mean follow-up period of 1.5 years. Conclusion: A “wait and see” strategy is justified in patients with non-progressive recurrent or residual cerebellar LGG after primary tumor resection (23.30) radiotherapy can be considered