Evaluation of cranial neuron number and their vacuolization in 129Sv SOD1G93A mice and non-transgenic littermates.

<p>Facial (A-E) and trigeminal (F-J) nuclei have been analyzed by Nissl (B-C, G-H) and H&E (D-E, I-J) staining to quantify brainstem neurons and to detect vacuolization, respectively. (A) Neuronal cell count of facial nucleus of 129Sv SOD1G93A mice at the advanced stage of the disease and relative non-transgenic littermates. (B-C) Nissl staining microphotographs of the facial nucleus of non-transgenic (Ntg, B) and SOD1G93A (C) 129Sv mice at the advanced stage of the disease. (D-E) H&E staining microphotographs of the facial nucleus of non-transgenic (Ntg, D) and SOD1G93A (E) 129Sv mice at the advanced stage of the disease. (F) Neuronal cell count of trigeminal nucleus of 129Sv SOD1G93A mice at the advanced stage of the disease and relative non-transgenic littermates. (G-H) Nissl staining microphotographs of the trigeminal nucleus of non-transgenic (Ntg, G) and SOD1G93A (H) 129Sv mice at the advanced stage of the disease. (I-J) H&E staining microphotographs of the trigeminal nucleus of non-transgenic (Ntg, I) and SOD1G93A (J) 129Sv mice at the advanced stage of the disease. Scale bar represents 100 μm in Nissl microphotographs (B-C, G-H) and 200 μm in H&E microphotographs (D-E, I-J). AS = advanced stage. The total neuron number was obtained multiplying the raw counts by four to give an estimate of the total cell number in the nucleus. Data are expressed as mean ± SD. Statistical analysis: Student’s t-test. ** p-value<0.01 between SOD1G93A and respective Ntg littermates. 4 animals for each group.</p

Two-Stage vs Single-Stage Thermophilic Anaerobic Digestion: Comparison of Energy Production and Biodegradation Efficiencies

Two-stage anaerobic digestion (AD) for integrated biohydrogen and biomethane production from organic materials has been reported to promise higher process efficiency and energy recoveries as compared to traditional one-stage AD. This work presents a comparison between two-stage (reactors R1 and R2) and one-stage (reactor R3) AD systems, fed with identical organic substrates and loading rates, focusing the attention on chemical and microbiological aspects. Contrary to previous experiences, no significant differences in overall energy recovery were found for the two-stage and one-stage AD systems. However, an accumulation in R2 of undegraded intermediate metabolites (volatile fatty acids, ketones, amines, amino acids, and phenols) was observed by GC-MS. These compounds were thought to be both cause and effect of this partial inefficiency of the two-stage system, as confirmed also by the less diverse, and thereby less efficient, population of fermentative bacteria observed (by PCR-DGGE) in R2. The extreme environment of R1 (low pH and high metabolites concentrations) probably acted as selector of metabolic pathways, favoring H₂-producing bacteria able to degrade such a wide variability of intermediate metabolites while limiting other strains. Therefore, if two-stage AD may potentially lead to higher energy recoveries, further efforts should be directed to ensure process efficiency and stability

Disease progression in C57 and 129Sv SOD1G93A mice.

<p>Grip strength measurements of C57 SOD1G93A (A) and 129Sv SOD1G93A (B) mice analysed in vivo for T2 relaxation time. Time points of MRI and histological studies are reported as arrows in correspondence to the respective ages of analysis: ON = Onset, SS = symptomatic, AS = advanced stage. (C) Disease onset of C57 SOD1G93A (15.6 ± 0.8) and 129Sv SOD1G93A mice (13.8 ± 0.4) analysed with T2 MRI. Statistical analysis: Student’s t-test. ** = p-value<0.01. Data are expressed as mean ± SD. N = 6 animals for C57 SOD1G93A mice and 5 animals for 129Sv SOD1G93A mice.</p

T2 relaxation time of brainstem nuclei of SOD1G93A mice at different stages of the disease.

<p>(A) Representative T2-weighted images are shown from a SOD1G93A (G93A) and a non-transgenic (Ntg) mouse. The ROIs correspondent to facial nuclei are indicated by a white arrow, to the trigeminal nuclei are indicated by a white arrowhead and to the inferior collliculi are indicated by a black arrow. T2 relaxation times of facial (B, E) and trigeminal (C, F) nuclei and of inferior colliculus (D, G) have been analyzed in C57 (B, C, D) and 129Sv (E, F, G) mice. SOD1G93A and non-transgenic mice have been compared during the progression of the disease for each strain. EPS = early pre-symptomatic stage, PS = pre-symptomatic stage, ON = Onset, SS = Symptomatic stage, AS = advanced stage. Data are expressed as mean ± SD. Statistical analysis: Two-way ANOVA, followed by Bonferroni’s post-hoc test. Facial nucleus: in C57 mice genotype [F (1,7) = 623.8, p<0.0001], stage [F (4,28) = 52.82, p<0.0001] and interaction [F (4,28) = 52.86, p<0.0001], while in 129Sv genotype [F (1,6) = 14.52, p = 0.0089], stage [F (3,18) = 2.481, p = 0.0939] and interaction [F (3,18) = 2.869, p0.0652]. Trigeminal nucleus: in C57 mice genotype [F (1,7) = 72.81, p<0.0001], stage [F (4,28) = 15.99, p<0.0001] and interaction [F (4,28) = 18, p<0.0001]. Inferior Colliculus: genotype [F (1,7) = 0.5181, p = 0.495], stage [F (4,28) = 2.329, p = 0.0806] and interaction [F (4,28) = 0.4251, p = 0.7892], while in 129Sv genotype [F (1,6) = 4.081, p = 0.0899], stage [F (3,18) = 0.5146, p = 0.6774] and interaction [F (3,18) = 3.294, p = 0.0443]. Stars indicate: * = p-value<0.05, ** = p-value<0.01, # = p-value<0.001. 6–5 animals in the SOD1G93A and 3 animals in the Ntg groups.</p

Diffusion Tensor Imaging (DTI) parameters and axonal count of lumbar spinal cord in C57 and 129Sv SOD1G93A mice during disease progression.

<p>(A) On the left, representative images of the Fractional Anisotropy of the lumbar spinal cord have been reported. Different white matter regions were took into consideration for the analyses: ventro-medial (vmWM), ventro-lateral (vlWM), dorso-lateral (dlWM) and dorsal (dWM). White lines represent the white matter area taken into consideration. Different diffusion parameters have been considered for the analysis of white matter degeneration through DTI: Fractional Anisotropy (a,c,e,g) and axial diffusivity (b,d,f,h). (B) The axonal count has been performed in the same white matter regions analysed with DTI. SOD1G93A mice have been analyzed at the onset, symptomatic (Sympto) and advanced stage (AS) of the disease. Data are expressed as ratio between SOD1G93A and their respective Ntg mice. Mean ± SD is reported. Statistical analysis: Two-way ANOVA has been used to compare Ntg and SOD1G93A mice during the disease progression, followed by Bonferroni’s post-hoc test. Stars indicate: * p-value<0.05, ** p-value<0.01, *** p-value<0.001, **** p-value<0.0001, between SOD1G93A and their respective Ntg controls of the same mouse strain. # indicates p-value<0.05 between C57 and 129Sv mice of the same disease stage. 4–3 animals for each group.</p