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Objective To assess frequency, types and long-term outcome of neurological manifestations during M. pneumoniae infection in paediatric patients. Methods Hospital charts of patients with IgM titer to M. pneumoniae hospitalized at our institution between January 2000 and December 2013 were analyzed retrospectively. Patients with/without neurological manifestations were compared. Possible risk factors were addressed by uni- and multivariate analyses. Patients with neurological symptoms were asked to take part in an evaluation of long-term outcome by standardized telephone interview to assess the Expanded Disability Status Score (EDSS), following written informed consent. Results 89 patients with acute M. pneumoniae infection were identified. Neurological symptoms were described in 22/89 (11 f and 11 m) patients with a median age of 11 y (4.8–15.7), including ataxia (9/22), meningism (6/22), impaired consciousness (9/22), seizures (4/22), impaired vision (7/22), speech disorders (5/22), paresis (2/22), muscle weakness (4/22), paraesthesia (5/22), and headaches or dizziness (10/22). Neurological syndromes included encephalitis (6/22), meningitis (2/22), cerebellitis (1/22), transvers myelitis (1/22), vestibular neuritis (1/22), and narcolepsy (1/22). CSF analysis obtained in 17/22 patients showed elevated cell count in 8/17 and protein in 3/17. EEG was obtained in 11 showing generalized slowing in 5, focal slowing in 3, and epileptic discharge in 1. Age <12 y was a risk factors for neurological symptoms. Lower respiratory tract symptoms and pre-existing immune deficit were associated with a reduced risk for neurological manifestations in our cohort. Data on long term follow-up could be obtained in 18/22 patients after a median of 5.1 y (0.6–13). In 8/18, parents reported ongoing neurological deficits during the EDSS interview. Conclusion Neurological symptoms occur in 25% of hospitalized paediatric patients with M. pneumoniae infection, especially in young children. Outcome is often favorable, but significant sequels are reported by 44%. [ABSTRACT FROM AUTHOR]

Mesenchymal stromal cells (MSCs) have therapeutic potential for a variety of diseases; however, preclinical effectiveness has not consistently been replicated in human trials. Rapidly thawed cryopreserved MSCs are often used in clinical trials as such products are more feasible from a logistic and cost perspective. However, recent reports suggest compromised MSC function in freshly thawed cells and this may result in diminished efficacy. To improve MSC function post-thaw, we tested a new class of cryoprotectants that can inhibit ice recrystallization and protect MSCs against cryoinjury. More specifically, we document the effect of two small-molecule ice recrystallization inhibitors (IRIs) N -(2-fluorophenyl)-D-gluconamide (2FA) and p -bromophenyl-β-D-glucopyranoside (pBrPh-Glc). Donor MSCs previously cryopreserved in passage 1 in Master cell banks were thawed and cultured in xeno- and serum-free media for 4 days. After 4 days in culture, MSCs were lifted, counted and cryopreserved at 2 × 106 cells/mL in Standard Freezing Media (SFM, 5% Human Albumin and 10% DMSO in PlasmaLyte A) with or without IRIs (15mM 2FA or 30mM pBrPh-Glc) using an optimized step-down program. Cryovials were subsequently thawed and cells were assessed for viability, identity and recovery post-thaw. Additionally, cell proliferation and interferon-gamma induced IDO expression were measured as surrogate markers of recovery and potency post-thaw. MSCs cryopreserved in SFM had a recovery of 80% and viabilities of 92.5% and 82.3% as measured by Trypan Blue exclusion and Annexin V and PI respectively. In comparison, MSCs cryopreserved in SFM containing 2FA or pBrPh-Glc had recoveries of 91.3 ± 0.7% or 73.3 ± 6.7% respectively. Cell viability as measured by Trypan blue exclusion was 90.9 ± 0.9% in the presence of 2FA versus 87.6 ± 0.8% for pBrPh-Glc. Significantly higher proliferation rates were observed 24h post-thaw in SFM containing 2FA (23.6% ± 1.9% cells in S-phase) than pBrPh-Glc (5.6% ± 0.5%) or SFM alone (6.88%). Additionally, MSCs cultured in XF-and SF-media had lower doubling times (27.4h ± 3.6h versus 59.7h ± 5.3h) and underwent more population doublings (2.71 ± 0.32 versus 1.22 ± 0.10) when cryopreserved in SFM containing 2FA. This effect was enhanced when cells were seeded at very low densities of 100 and 250 cells/cm2. In conclusion, we demonstrate marked improvement in post-thaw MSC recovery, viability and function when cryopreserved with the ice recrystallization inhibitor 2FA. [ABSTRACT FROM AUTHOR]