Oral supplementation with Scabiosa artropurperea L. aqueous extract improves the in vivo sexual behaviour, sperm quality and androgen level in rams.

The effects of an aqueous extract of Scabiosa atropurpurea L. (AES) on the reproduction potential of Queue Fine de l'Ouest rams were evaluated over 9 weeks. Eighteen mature (4-6 years old) rams (52.8 ± 2.6 kg) were divided into three groups. The control (C) group was fed oat hay ad libitum with 700 g of concentrate and the other two groups were fed the same diet supplemented with AES at 1 and 2 mg/kg body weight (AES1 and AES2, respectively). Ram sperm was collected with an artificial vagina (2 × 2 days/week) to evaluate sperm production and quality, antioxidant activity, the adenosine triphosphate (ATP) and calcium concentrations. Sexual behaviour and plasma testosterone concentrations were also investigated. The administration of AES improved sexual behaviour (the duration of contact and the number of lateral approaches). The addition of AES also improved individual spermatozoa motility (C: 71.7% ± 6.3%; AES1: 78.3% ± 4.9%; AES2: 83.8% ± 4.4%), the sperm concentration (C: 5.6 ± 0.36; AES1: 6.4 ± 0.81; AES2: 6.7 ± 0.52 × 109 spermatozoa/mL), the ATP ratio (C: 1 ± 0.08; AES1: 2.1 ± 0.08; AES2: 3.3 ± 0.08) and the calcium concentration (C: 5.6 ± 0.24; AES1: 7.7 ± 0.21; AES2: 8.1 ± 0.24 mmol/L). AES treatment decreased the percentage of abnormal sperm (C: 18.5% ± 1.2%; AES1: 16.2% ± 1.1%; AES2: 14.8% ± 0.94%) and DNA damage (C: 62%; AES1: 27%; AES2: 33%) and was associated with elevated seminal fluid antioxidant activity (C: 22 ± 0.27; AES1: 27.1 ± 1.08 and AES2: 27.5 ± 0.36 mmol Trolox equivalents/L) and plasma testosterone (C: 8.3 ± 0.7; AES1: 11.7 ± 0.4; AES2: 15 ± 0.7 ng/L). In conclusion, our study suggests that S. atropurpurea may be potentially useful to enhance libido and sperm production and quality in ram.

The Influence of Cathode Degradation Products on the Anode Interface in Lithium-Ion Batteries.

Degradation of cathode materials in lithium-ion batteries results in the presence of transition metal ions in the electrolyte, and these ions are known to play a major role in capacity fade and cell failure. Yet, while it is known that transition metal ions migrate from the metal oxide cathode and deposit on the graphite anode, their specific influence on anode reactions and structures, such as the solid electrolyte interphase (SEI), is still quite poorly understood due to the complexity in studying this interface in operational cells. In this work we combine operando electrochemical atomic force microscopy (EC-AFM), electrochemical quartz crystal microbalance (EQCM), and electrochemical impedance spectroscopy (EIS) measurements to probe the influence of a range of transition metal ions on the morphological, mechanical, chemical, and electrical properties of the SEI. By adding representative concentrations of Ni2+, Mn2+, and Co2+ ions into a commercially relevant battery electrolyte, the impacts of each on the formation and stability of the anode interface layer is revealed; all are shown to pose a threat to battery performance and stability. Mn2+, in particular, is shown to induce a thick, soft, and unstable SEI layer, which is known to cause severe degradation of batteries, while Co2+ and Ni2+ significantly impact interfacial conductivity. When transition metal ions are mixed, SEI degradation is amplified, suggesting a synergistic effect on the cell stability. Hence, by uncovering the roles these cathode degradation products play in operational batteries, we have provided a foundation upon which strategies to mitigate or eliminate these degradation products can be developed.

Analysis of Management Practices and Breeders' Perceptions of Climate Change's Impact to Enhance the Resilience of Sheep Production Systems: A Case Study in the Tunisian Semi-Arid Zone.

Global climate change inflicts unambiguous risks on agricultural systems and food security. Small ruminants are known for their ability to adapt to changing environmental conditions. This paper aims to characterize sheep production systems in a vulnerable agricultural zone and the breeders' perceptions of climate change to apprehend challenges that they are confronting, and formulate resilience actions. The data analysis is based on 94 semi-structured surveys of sheep farmers carried out in the Tunisian semi-arid region. The PCA analysis results revealed three main sheep production systems. The agro-pastoral rain-fed system (AGPRF) is dominant (55%), with large farms and common pastures integrating cereals and fodder. The agro-pastoral irrigated system (AGPI: 20%) is characterized by small-area and forage irrigation (1.8 ha) and a smaller number of ewes but a greater use of animal feed supplementation. The agro-sylvo-pastoral system (AGSP: 25%) is a system where grazing is based on common lands and using tree sub-products, while the agricultural area is exclusively used to cultivate cereal crops. Sheep breeders' climate perceptions are summarized as unpredictable climate events, a decrease in precipitation, and an increase in temperature. Resilience actions principally consist of reducing flocks' numbers, using alternative local feed, fodder, and water resources, and building more shelters and planting more trees in the grazing areas. Nevertheless, cost-effectiveness should be considered in such vulnerable zones to insure the sheep production systems' sustainability.

Impact of Body Reserves Dynamic on Productivity and Reproductive Performance in Fat-Tail and Thin-Tail Sheep Breeds over Contrasting Production Cycles.

Under climate change, animal breeding faces severe nutritional challenges, exhibiting their resilience and efficiency to survive and produce. The animals' robustness can be evaluated through the use and reconstitution of body reserves (BR). This study investigated the dynamics of ewes' BR, measured through body weight (BW) and the body condition score (BCS), and their effect on production performances of 480 ewes belonging to fat-tailed Barbarine (BB) and thin-tailed Queue Fine de l'Ouest (QFO) ewes over two consecutive years. The BW and BCS were recorded across physiological stages. Reproductive parameters and lambs' growth were calculated. For both years and breeds, the decrease in the BCS between mating and lambing showed BR mobilization to balance nutritional restrictions, which continues until suckling; after weaning, a BR accretion was noted. The lambs' birth weight was similar regardless of the dams' BCS at lambing, though it was lower in the second year (3.5 vs. 4 kg). Lambs' growth parameters depended on ewes' BCS at lambing and weaning. Fertility rates changed with BCS changes and the higher values (80%) were recorded for ewes with BCSs between 2 and 2.5. However, prolificacy was similar for both breeds and years (p > 0.05). In conclusion, ewes have developed adaptative capacities through BR mobilization to cope with environmental challenges and successfully rear their offspring. To maintain a high offspring growth rate, it is recommended to plan correct flock nutrition during suckling.

Comparative Study of Carcass Characteristics and Meat Quality of Local Mediterranean Donkey Breeds.

This study aimed to evaluate carcass and meat quality traits in Masri (n = 14) and North African (n = 14) male donkeys, raised in a semi-intensive breeding system, grazing on mountainous areas, with supplementation of 1 kg of barley per day per animal, slaughtered at 5 (±0.48) years old. Compared to Masri, the North African population exhibited higher (p < 0.05) final body weights (181.7 ± 2.1 and 212.5 ± 7.6 kg) and cold carcass weights (101.7 ± 1.3 and 116.2 ± 4.5 kg), but lower (p < 0.05) cold dressing percentages (56.0 ± 0.4 and 54.6 ± 0.4%). Meat quality analyses showed higher (p < 0.05) cooking loss values in Masri meat (43.9 ± 0.8 vs. 39.9 ± 1.2%). Among the meat color parameters, the hue value was higher (p < 0.05) in North African samples (0.42 ± 0.01 vs. 0.39 ± 0.1). Meat from North African donkeys had higher (p < 0.05) dry matter, fat, and protein contents. Meat amino acid analysis revealed abundant levels of lysine, leucine, and methionine, in both populations. Donkey meat from both populations presented a high polyunsaturated fatty acids content, resulting in polyunsaturated fatty acids/saturated fatty acids and omega 6/omega 3 ratios for all breeds close to the recommended values for human health. Atherogenic and thrombogenic indices were also suggested to have positive effects on consumers' artery health. The characteristics of donkey meat present intriguing nutritional aspects compared to ruminant meat, and its production should be encouraged in the rural development of inner-Mediterranean areas.

Investigating the mechanism of phosphorene nanoribbon synthesis by discharging black phosphorus intercalation compounds.

Phosphorene nanoribbons (PNRs) can be synthesised in intrinsically scalable methods from intercalation of black phosphorus (BP), however, the mechanism of ribbonisation remains unclear. Herein, to investigate the point at which nanoribbons form, we decouple the two key synthesis steps: first, the formation of the BP intercalation compound, and second, the dissolution into a polar aprotic solvent. We find that both the lithium intercalant and the negative charge on the phosphorus host framework can be effectively removed by addition of phenyl cyanide to return BP and investigate whether fracturing to ribbons occurred after the first step. Further efforts to exfoliate mechanically with or without solvent reveal that the intercalation step does not form ribbons, indicating that an interaction between the amidic solvent and the intercalated phosphorus compound plays an important role in the formation of nanoribbons.

Evaluation of acute toxicity of Scabiosa artropurperea var.maritima aqueous extracts in Swiss mice.

Scabiosa artropurperea var.maritima is a plant widely distributed in the Mediterranean region and used as a traditional medicine. The present study evaluated the biochemical composition and the potential toxicity of aqueous extract of whole Scabiosa artropurperea var.maritima through acute toxicity oral administration in male mice. Phytochemical analysis of the Scabiosa artropurperea var.maritima revealed high levels of reductor sugars and significant flavonoid and total phenol content. The aqueous extract of Scabiosa artropurperea var.maritima was daily oral administered to mice at doses of 300 (group 1), 2000 (group 2) and 4000 (group 3) mg/kg body weight per day for 14 days. We observed no significant difference in the consumption of food, body weight and relative organ weights except for an increase in the seminal vesicles weight in group 3. Hematological parameters revealed the non-adverse effects of prolonged oral consumption of Scabiosa artropurperea var.maritima except for a slight increase but significant of percentage of hematocrit in group 1 and 3 and a decrease in percentage of granulocytes in group 2. The histopathologic examination did not show any differences in vital organs. We also observed non-adverse effects on the reproductive parameters including testosterone concentration, spermatozoa motility and morphologies. Based on our findings, the aqueous extract of Scabiosa artropurperea var.maritima could be considered safe for oral medication in animals.

Quality indicators of public maternity units in the governorate of Monastir (Tunisia).

INTRODUCTION: Increasing access to healthcare for expectant mothers is a national goal. In Monastir, Tunisia, some Peripheral Maternity Units (PMUs) required assessment. Our goals were to describe the delivery activities in MUs (maternity units) and to assess whether some of PMUs need to have their activities replaced. METHOD: We analyzed aggregate data of deliveries in Monastir from 2015 to 2020. The gouvernorate's seven public MUs were included. Only the morning activity was allotted for obstetricians and gynecologists, in RMUs 1 and 2, whereas they were not available in all PMUs. Data was gathered from the reports of the National Perinatal Program. Both the availability of Comprehensive Essential Obstetric Care (CEOC) and Basic Essential Obstetric Care (BEOC) were calculated. Trends were calculated using Joinpoint software. The Annual Percent Change (APC) was calculated. RESULTS: The number of births decreased from 2015 to 2020 (APC= -4.3%: 95%CI : -6; -2.4; p = 0.003). The largest significant decreases in APCs of deliveries were reported in PMU 2 (APC = -12.6% (95%CI : -20; -4.4; p = 0.014), in PMU 3 (APC = -29.3% (95%CI : -36.5; -21.4; p = 0.001), and in PMU 4 (APC = -32.9% (95%CI: -49.1; -11.5); p = 0.016). If PMU 3 and 4 were no longer operating as maternity facilities, BEOC and CEOC standards would still be adequat. For accessibility, both PMU 3 and PMU 2 are accessible from PMU 4 and PMU 1, respectively. CONCLUSIONS: Pregnant women prefer to give birth in obstetric services with ability to perform emergency caesarean at the expense of PMU. Nowadays, it appears that accessibility is less important than the presence of qualified human resources when a pregnant woman choose a maternity hospital.

Electrode Materials for Enhancing the Performance and Cycling Stability of Zinc Iodide Flow Batteries at High Current Densities.

Aqueous redox flow battery systems that use a zinc negative electrode have a relatively high energy density. However, high current densities can lead to zinc dendrite growth and electrode polarization, which limit the battery's high power density and cyclability. In this study, a perforated copper foil with a high electrical conductivity was used on the negative side, combined with an electrocatalyst on the positive electrode in a zinc iodide flow battery. A significant improvement in the energy efficiency (ca. 10% vs using graphite felt on both sides) and cycling stability at a high current density of 40 mA cm-2 was observed. A long cycling stability with a high areal capacity of 222 mA h cm-2 is obtained in this study, which is the highest reported areal capacity for zinc-iodide aqueous flow batteries operating at high current density, in comparison to previous studies. Additionally, the use of a perforated copper foil anode in combination with a novel flow mode was discovered to achieve consistent cycling at exceedingly high current densities of >100 mA cm-2. In situ and ex situ characterization techniques, including in situ atomic force microscopy coupled with in situ optical microscopy and X-ray diffraction, are applied to clarify the relationship between zinc deposition morphology on the perforated copper foil and battery performance in two different flow field conditions. With a portion of the flow going through the perforations, a significantly more uniform and compact zinc deposition was observed compared to the case where all of the flow passed over the surface of the electrode. Results from modeling and simulation support the conclusion that the flow of a fraction of electrolyte through the electrode enhances mass transport, enabling a more compact deposit.

Substitution of soyabean meal by faba bean improves body weight, sperm production and testosterone concentration in mature Queue Fine de l'Ouest rams.

The current study aimed at investigating the effects of partial or total substitution of soybean meal with faba bean on reproductive parameters of Queue Fine de l'Ouest rams. Eighteen adult rams (49.8 ± 3.7 kg and 2.4 ± 1.5 years old) were divided into three homogenous groups. Rams received oat hay ad libitum and 3 types of concentrate (33 g/BW0.75) where soybean meal was the main protein source (SBM diet, n = 6), partially (50%, SBMFB diet, n = 6) or totally (100% FB diet, n = 6) substituted in nitrogen basis by local faba bean. The volume of ejaculate, sperm concentration and sperm mortality rate were measured weekly by collecting semen using an artificial vagina. Serial blood samples were collected 30 days and 120 days after the beginning of the experiment in order to assess plasma testosterone concentrations. The results showed that hay intake was affected (P < 0.05) by the nature of the nitrogen source incorporated (1032.3 ± 12.2; 1026.8 ± 56.6 and 972.8 ± 39.05 g DM/d for SBM, FB and SBMFB respectively. The average live weight of the rams increased from 49.8 ± 0.4 (week 1) to 57.3 ± 0.9 kg (week 17) without effect of the diet. A positive effect of faba bean incorporation in the concentrate was observed on ejaculate volume, concentration and spermatozoa production. All these parameters were significantly higher in SBMFB and FB groups than in SBM group (P < 0.05). The protein source did not affect the percentage of dead spermatozoa and the total abnormalities which were similar for the three diets (38.7; 35.8 and 38.1% for SBM; SBMFB and FB, respectively). The mean testosterone concentration was higher (P < 0.05) in rams fed with faba bean (1.7 ± 0.7 to 1.9 ± 0.7 ng/ml for the SBMFB and the FB groups respectively) than in rams fed SBM diet (1.06 ± 0.5 ng/ml). It was concluded that the substitution of soybean meal by faba bean improved the reproductive performance and did not affect sperm quality in Queue Fine de l'Ouest rams.

Black Phosphorus Degradation during Intercalation and Alloying in Batteries.

Numerous layered materials are being recognized as promising candidates for high-performance alkali-ion battery anodes, but black phosphorus (BP) has received particular attention. This is due to its high specific capacity, due to a mixed alkali-ion storage mechanism (intercalation-alloying), and fast alkali-ion transport within its layers. Unfortunately, BP based batteries are also commonly associated with serious irreversible losses and poor cycling stability. This is known to be linked to alloying, but there is little experimental evidence of the morphological, mechanical, or chemical changes that BP undergoes in operational cells and thus little understanding of the factors that must be mitigated to optimize performance. Here the degradation mechanisms of BP alkali-ion battery anodes are revealed through operando electrochemical atomic force microscopy (EC-AFM) and ex situ spectroscopy. Among other phenomena, BP is observed to wrinkle and deform during intercalation but suffers from complete structural breakdown upon alloying. The solid electrolyte interphase (SEI) is also found to be unstable, nucleating at defects before spreading across the basal planes but then disintegrating upon desodiation, even above alloying potentials. By directly linking these localized phenomena with the whole-cell performance, we can now engineer stabilizing protocols for next-generation high-capacity alkali-ion batteries.

Engineering Solution-Processed Non-Crystalline Solid Electrolytes for Li Metal Batteries.

Non-crystalline Li-ion solid electrolytes (SEs), such as lithium phosphorus oxynitride, can uniquely enable high-rate solid-state battery operation over thousands of cycles in thin film form. However, they are typically produced by expensive and low throughput vacuum deposition, limiting their wide application and study. Here, we report non-crystalline SEs of composition Li-Al-P-O (LAPO) with ionic conductivities > 10-7 S cm-1 at room temperature made by spin coating from aqueous solutions and subsequent annealing in air. Homogenous, dense, flat layers can be synthesized with submicrometer thickness at temperatures as low as 230 °C. Control of the composition is shown to significantly affect the ionic conductivity, with increased Li and decreased P content being optimal, while higher annealing temperatures result in decreased ionic conductivity. Activation energy analysis reveals a Li-ion hopping barrier of ≈0.4 eV. Additionally, these SEs exhibit low room temperature electronic conductivity (< 10-11 S cm-1) and a moderate Young's modulus of ≈54 GPa, which may be beneficial in preventing Li dendrite formation. In contact with Li metal, LAPO is found to form a stable but high impedance passivation layer comprised of Al metal, Li-P, and Li-O species. These findings should be of value when engineering non-crystalline SEs for Li-metal batteries with high energy and power densities.

Clinical and genetic spectra of 1550 index patients with hereditary spastic paraplegia.

Hereditary spastic paraplegia refers to rare genetic neurodevelopmental and/or neurodegenerative disorders in which spasticity due to length-dependent damage to the upper motor neuron is a core sign. Their high clinical and genetic heterogeneity makes their diagnosis challenging. Multigene panels allow a high-throughput targeted analysis of the increasing number of genes involved using next-generation sequencing. We report here the clinical and genetic results of 1550 index cases tested for variants in a panel of hereditary spastic paraplegia related genes analysed in routine diagnosis. A causative variant was found in 475 patients (30.7%) in 35/65 screened genes. SPAST and SPG7 were the most frequently mutated genes, representing 142 (9.2%) and 75 (4.8%) index cases of the whole series, respectively. KIF1A, ATL1, SPG11, KIF5A and REEP1 represented more than 1% (>17 cases) each. There were 661 causative variants (382 different ones) and 30 of them were structural variants. This large cohort allowed us to obtain an overview of the clinical and genetic spectrum of hereditary spastic paraplegia in clinical practice. Because of the wide phenotypic variability, there was no very specific sign that could predict the causative gene, but there were some constellations of symptoms that were found often related to specific subtypes. Finally, we confirmed the diagnostic effectiveness of a targeted sequencing panel as a first-line genetic test in hereditary spastic paraplegia. This is a pertinent strategy because of the relative frequency of several known genes (i.e. SPAST, KIF1A) and it allows identification of variants in the rarest involved genes and detection of structural rearrangements via coverage analysis, which is less efficient in exome datasets. It is crucial because these structural variants represent a significant proportion of the pathogenic hereditary spastic paraplegia variants (∼6% of patients), notably for SPAST and REEP1. In a subset of 42 index cases negative for the targeted multigene panel, subsequent whole-exome sequencing allowed a theoretical diagnosis yield of ∼50% to be reached. We then propose a two-step strategy combining the use of a panel of genes followed by whole-exome sequencing in negative cases.

Evidence of mosaicism in SPAST variant carriers in four French families.

Hereditary spastic paraplegias (HSP) are heterogeneous disorders, with more than 70 causative genes. Variants in SPAST are the most frequent genetic etiology and are responsible for spastic paraplegia type 4 (SPG4). Age at onset can vary, even between patients from the same family, and incomplete penetrance is described. Somatic mosaicism is extremely rare with only three patients reported in the literature. We report here SPAST mosaic variants in four unrelated patients. We confirm that mosaicism in SPAST is a very rare event with only four identified cases on more than 300 patients with a SPAST variant previously described by our clinical diagnostic laboratory.

Carcass Traits and Meat Quality of Fat-Tailed Lambs Fed Rosemary Residues as a Part of Concentrate.

Facing climate change implications on feeds unavailability, unconventional resources are being considered with a growing interest such as aromatic plant distillation residues with a two-fold object, enhancing meat quality by increasing the antioxidant properties and reducing feed prices which are often imported though expensive. Hence, this study aims to assess the effects of rosemary distillation residues (RR) incorporation in concentrate associated to two nitrogen sources as a substitute for standard concentrate on lamb's growth, carcass traits and meat quality. For this, 24 Barbarine male lambs (3 months old, 17.83 ± 2.6 kg body weight) were divided into three groups. All lambs received individually 600 g of oat hay as roughage and 600 g of standard concentrate for control group, 600 g of concentrate based on RR and soybean meal for RRS group and 600 g of concentrate based on RR and faba bean for RRF group. After 65 days of experiment, all lambs were slaughtered. Phenolic and tocopherol intakes were significantly higher for both RR groups compared to control (p < 0.05). Growth, carcass weights, dressing percentages and non-carcass component weights were unaffected by the diet (p > 0.05). Moreover, regional and tissular compositions and meat physical properties were similar irrespective of the diet (p > 0.05). All color parameters were similar among groups (p > 0.05). However, meat produced by lambs receiving RR-based concentrate was richer on vitamin E and polyphenol contents than control lambs (p < 0.05). Rosemary by-products may substitute the standard concentrate resulting in similar lamb's growth and carcass traits, while improving meat quality by increasing vitamin E content, which could improve its antioxidant power.

RNF170-Related Hereditary Spastic Paraplegia: Confirmation by a Novel Mutation.

BACKGROUND: Spastic paraparesis and biallelic variants functionally characterized as deleterious in the RNF170 gene have recently been reported by Wagner et al. 2019, strongly supporting the involvement of this gene in hereditary spastic paraplegia. METHODS: Exome sequencing was performed on 6 hereditary spastic paraplegia families previously tested on an hereditary spastic paraplegia-specific panel. RESULTS: We describe here a novel hereditary spastic paraplegia family with 4 affected members carrying a homozygous p.(Tyr114*) stop gain variant in RNF170. CONCLUSIONS: We confirm the involvement of biallelic truncating variants in RNF170 in a novel form of hereditary spastic paraplegia. © 2020 International Parkinson and Movement Disorder Society.

BCG vaccination and tuberculosis prevention: A forty years cohort study, Monastir, Tunisia.

We aimed to describe incidence, trends of tuberculosis (TB) over 18 years and to evaluate the impact of the BCG vaccine after four decades of immunization program according to three protocols. We performed a cohort study including declared cases in Monastir from January 1, 2000 to December 31, 2017. We reported 997 cases of TB. The predominant site was pulmonarylocalization (n = 486). The age standardized incidence of pulmonary and lymph node TB per 100,000 inh were 5.71 and 2.57 respectively. Trends were negative for pulmonary TB (PTB) (b = - 0.82; r = -0.67; p<10-3) and positive for lymph node localization (b = 1.31; r = 0.63; p<10-3). We had not notified cases of HIV associated with TB. Crude incidence rate (CIR) of PTB per 100,000 inh was 8.17 in Non-Vaccinated Cohort (NVC) and 2.85 in Vaccinated Cohort (VC) (p < 0.0001). Relative risk reduction (RRR) of BCG vaccination was 65.1% (95%CI:57.5;71.4) for pulmonary localization and 65% (95%CI:55; 73) for other localizations. We have not established a significant RRR of BCG vaccination on lymph node TB. Protocol 3 (at birth) had the highest effectiveness with a RRR of 96.7% (95%CI: 86.6%; 99.2%) and 86% (95%CI:71%;91%) in patients with PTB and other localizations TB respectively. In Cox regression model the HR was 0.061 (95% CI 0.015-0.247) for PTB and 0.395 (95% CI 0.185-0.844) for other localizations TB in patients receiving protocol 3 compared to NVC. For lymph-node TB, HR was 1.390 (95% CI 1.043-1.851) for protocol 1 and 1.849 (95% CI 1.232-2.774) for protocol 2 compared to NVC. Depending on the three protocols, the BCG vaccine had a positive impact on PTB and other TB localizations that must be kept and improved. However, protocols 1 and 2 had a reverse effect on lymph node TB.

Spastic paraplegia due to SPAST mutations is modified by the underlying mutation and sex.

Hereditary spastic paraplegias (HSPs) are rare neurological disorders caused by progressive distal degeneration of the corticospinal tracts. Among the 79 loci and 65 spastic paraplegia genes (SPGs) involved in HSPs, mutations in SPAST, which encodes spastin, responsible for SPG4, are the most frequent cause of both familial and sporadic HSP. SPG4 is characterized by a clinically pure phenotype associated with restricted involvement of the corticospinal tracts and posterior columns of the spinal cord. It is rarely associated with additional neurological signs. However, both age of onset and severity of the disorder are extremely variable. Such variability is both intra- and inter-familial and may suggest incomplete penetrance, with some patients carrying mutations remaining asymptomatic for their entire life. We analysed a cohort of 842 patients with SPG4-HSP to assess genotype-phenotype correlations. Most patients were French (89%) and had a family history of SPG4-HSP (75%). Age at onset was characterized by a bimodal distribution, with high inter-familial and intra-familial variability, especially concerning first-degree relatives. Penetrance of the disorder was 0.9, complete after 70 years of age. Penetrance was lower in females (0.88 versus 0.94 in males, P = 0.01), despite a more diffuse phenotype with more frequent upper limb involvement. Seventy-seven per cent of pathogenic mutations (missense, frameshift, splice site, nonsense, and deletions) were located in the AAA cassette of spastin, impairing its microtubule-severing activity. A comparison of the missense and truncating mutations revealed a significantly lower age at onset for patients carrying missense mutations than those carrying truncating mutations, explaining the bimodal distribution of the age at onset. The age at onset for patients carrying missense mutations was often before 10 years, sometimes associated with intellectual deficiency. Neuropathological examination of a single case showed degeneration of the spinocerebellar and spinocortical tracts, as well as the posterior columns. However, there were numerous small-diameter processes among unusually large myelinated fibres in the corticospinal tract, suggesting marked regeneration. In conclusion, this large cohort of 842 individuals allowed us to identify a significantly younger age at onset in missense mutation carriers and lower penetrance in females, despite a more severe disorder. Neuropathology in one case showed numerous small fibres suggesting regeneration.