Background: Urinary tract infections (UTI) are considered as common facilitating factors along with other infections in triggering febrile seizures (FS). The main purpose of the study is to identify specific patterns of UTIs, using a combination of inflammatory biomarkers, in order to differentiate UTIs from other bacterial diseases associated with FS.

Method: This study included a number of 197 distinct FS events, from patients hospitalized in the Sibiu Pediatric Hospital, among which 10.2% were diagnosed with UTIs. Results: In one third of patients with UTIs symptoms were limited to fever and FS. Using Two-Step cluster analysis, a distinct inflammatory pattern has emerged: higher PDW (median value 9,65 fl), P-LCR (median value 14,45%), VTM (median value 10,40 fl), PCR (median value 74,00 g/L) and neutrophil-lymphocyte ratio (median value 3,64), associated mainly (85,7%) with bacterial respiratory infections. UTIs were highly unlikely in the patients with significantly increased PCR values and normal values of platelet indices.

Conclusion. Considering the nonspecific clinical picture of UTIs at an early age, in order to optimize the management of FS a fast diagnosis of UTI is mandatory. Our study suggests that analyzing the inflammatory biomarkers interlink (rather than individual parameters) could help identify UTI patients, even when oligosymptomatic.

Batten Disease is a fatal, lysosomal storage disorder characterized by cognitive and motor deficits, vision impairments, and seizures. Loss of vision is a hallmark of 10 of the 13 Batten Disease subtypes. Our group has pioneered AAV9 gene therapy treatments achieving widespread transduction of the brain and spinal cord. Two clinical trials are currently ongoing at Nationwide Children’s Hospital delivering this vector via cerebrospinal fluid (CSF) to the brain and spinal cord for treatment of the lethal neurodegenerative aspects of Batten Disease. However, AAV9 transduction of the retina after CSF delivery is limited and there is a critical need for a solution that prevents vision loss and further improves quality of life for Batten Disease patients.

Similar to most genetic ocular diseases, photoreceptor degeneration is the most commonly cited pathology in patients. However, recent studies suggest that in some subtypes of Batten Disease, expression must be rescued also within the deeper layers of the retina that are difficult to reach with therapeutic vectors (inner nuclear layer, INL). We performed single cell RNA sequencing of mice and non-human primate retinas in collaboration with Dr. Fischer (OSU) and concluded that Batten Disease vision specific gene therapy needs to target a wide range of cells, including the INL, which is a major challenge for translation of a vision specific therapy to the clinic.

We have recently discovered that administration of neuraminidase (NA), a sialidase enzyme, prior to or in combination with AAV9.GFP, drastically increases transduction throughout the murine C57Bl/6 retina including the INL and all the way through to the photoreceptor layer. Our preliminary data indicates GFP expression in almost all, if not all, retinal cell types using this method. Importantly, we have confirmed successful targeting of INL bipolar cells, a notoriously difficult cell-type to transduce, and up to 40% increase in Müller glia transduction. Preliminary histological examination indicates no damage or alterations in retinal integrity.

While additional testing in large animal models is required and scheduled to occur in a WT pig model in Autumn 2020, this remarkable discovery suggests that it may now be possible to target every cell-type of the retina with a single AAV vector. This is especially important in regard to treatment of Batten Disease but has many additional implications. Current retinal therapies that require a more invasive subretinal delivery to photoreceptors could now opt for the safer intravitreal delivery strategy. In addition, this strategy would be highly useful in the field of optogenetics, where investigators continue to struggle to find an efficient way to express light-sensitive opsins in cells of the INL to restore sight in individuals that have already lost their photoreceptors.

Noise-induced hearing loss (NIHL) is one of the most prevalent disabilities for which effective therapeutic treatment is currently lacking. Auditory injury caused by excessive or constant noise exposure damages the sensory elements of the ear, leading to metabolic or mechanical damage to hair cells and subsequent degeneration of spiral ganglion neurons (SGNs). To improve hearing in the NIHL patient population, maintenance and regeneration of inner hair cell-SGN synapses is critical. Although effective compounds for the treatment of NIHL have been suggested, challenges in delivery of the compounds to the inner hair cells (IHCs) hinder translation of these therapies to a clinical setting. Current routes of administration are ineffective in reaching IHCs or are invasive and may damage the cochlea. In this study, we examined less invasive delivery routes, via intrathecal (cerebrospinal fluid delivery) or intravenous injection, to administer adeno-associated virus serotype 9 (AAV9) expressing green fluorescent protein (GFP) to target IHCs and SGNs. To induce auditory injury, mice were exposed to 100 dB sound pressure level (SPL) octave band noise for 2 hours. Mice were then injected at various time points post noise injury to determine if noise exposure influenced targeting efficiency. Three weeks post injection, cochleae were processed and analyzed for GFP expression using immunofluorescence. Our results demonstrate that intravenous delivery was not successful in targeting of IHCs or SGNs at any time point measured. However, intrathecally injected AAV9 was highly effective in targeting cells in both healthy and noise-damaged cochleae at multiple time points post-noise exposure. We conclude that gene therapy techniques can be utilized to efficiently deliver therapeutic transgenes to the cochlea using cerebrospinal fluid as a delivery route. These findings determine a novel route of delivery and open a new avenue of therapeutic treatment for auditory injury and cochlear disorders.

Patient diversity and unknown disease cause are major challenges for drug development and clinical trial design for ALS. Moreover, the heterogeneity of the ALS patient population is not reflected in currently available transgenic animal models making direct translation of potential therapeutics difficult. To address this, we utilize direct conversion technology to transform skin biopsies from ALS patients into neuronal progenitor cells (NPC). Using induced astrocytes (iAs) differentiated from these NPCs in co-culture with mouse embryonic motor neurons, we developed an in vitro ALS model to screen potential therapeutics. We have screened numerous compounds on multiple sporadic (sALS) and familial (fALS, mtSOD1 and C9ORF72) iAs and observed a diverse patient response to different therapeutic agents. Here, we utilize the compound, CuATSM, to subgroup therapeutic responsiveness of patient lines and identified shared dysfunctional pathways between responders. Following detailed analysis of ALS disease markers, elevated mitochondrial activity states was present in all CuATSM responders and was nonexistent in nonresponders. Treatment of iAs with CuATSM restored this activity to healthy control levels. Together, these findings suggest that iAs can be used to identify both disease modifiers and pathways dysregulated in an individual. Thus enhanced understanding of cellular profiles could facilitate personalized medicine based treatments.

Various studies have concluded that in multiple sclerosis (MS) metabolic changes related to energy alterations are generated at the mitochondrial level. These changes produce loss of muscle mass and lead to high levels of oxidative stress related to low levels of the enzyme Paraoxonase 1 (PON1). The ketone bodies, mainly beta-hydroxybutyrate (BHB), restore this energetic alteration by improving muscular activity.

This pilot study was conducted over 4 months with 51 patients diagnosed with MS, randomly divided into: an intervention group that received an isocaloric diet of a Mediterranean origin supplemented with 60 ml of coconut oil; and a control group that received the same base diet but without coconut oil. Before and after treatment, anthropometric measurements and blood samples were taken, measuring serum PON1 and BHB concentrations, to determine the role of PON1 in muscle improvement after increased blood BHB in MS patients.

A significant increase in PON1 was obtained, associated with an increase in lean mass and an increase in BHB, so it was concluded that this enzyme appeared to be a good marker of decreased oxidation status in MS patients showing muscle improvement after increased BHB in blood.