Raynaud’s phenomenon in undifferentiated connective tissue disease (UCTD)

The aim of this study was to ascertain which clinical and immunological factors are associated with Raynauds phenomenon (RP) in patients with undifferentiated connective tissue disease (UCTD) and to investigate microvascular involvement. A total of 78 patients were evaluated. They all showed symptoms suggestive of a connective tissue disorder (CTD), but did not fulfil the criteria for any of the defined CTDs. They all had a disease duration of at least 1 year. Nailfold capillaroscopy (NC) was performed using a computerised videomicroscope. We diagnosed RP in 52.5% of our patients. Patients with RP showed a higher occurrence of oesophageal dysmotility (p=0.001) and anti-ribonucleoprotein (RNP) antibodies (p=0.004) than those without RP. The distinguishing capillaroscopic characteristics of UCTD patients with RP were widened and irregularly enlarged loops (75 and 55%, respectively), giant capillaries (35%), and less than two haemorrhages per finger (40%). The combination of features indicative of a slow scleroderma pattern was present in 18 of 40 patients with UCTD and RP (p=0.0003). Only 3 of the original 78 patients (3.8%) developed a definite CTD. In none of our patients did we observe avascular areas or changes from the original capillaroscopic pattern during follow-up examination. Our study indicates that patients with UCTD would seem to have a benign form of RP, since they show the absence of cutaneous complications, the existence of a mild microvascular damage and a stable nailfold capillary pattern. Further examinations of these patients will be required in order to confirm our findings.     

Morphometric investigations of the mitochondrial damage in ceroid lipopigment accumulation due to vitamin E deficiency

Numeric (Nv) and volume (Vv) densities, as well as the average size (skeleton: Sk) of synaptic mitochondria from adult, normally fed and adult, vitamin E deficient animals (11 months of age) were semiautomatically measured by computer-assisted morphometry in the cerebellar granular layer. Nv, Vv and the average mitochondrial volume (V) were measured on perikaryal Purkinje cell organelles preferentially stained for succinic dehydrogenase (SDH) activity. Adult vitamin E deficient animals showed a significant decrease of Nv, a significant increase of Sk and an unchanged value of Vv. While in adult normally fed animals the mitochondria of increased size (Sk>5 microm) were 5.3%, in the adult vitamin E deficient rats this fraction accounted for 25.5%. In Purkinje cell perikarya, vitamin E deficiency resulted in a significant decrease of Vv, Nv and V, as well as a steeper reduction of the percentage of SDH-positive mitochondria of larger size. Taken together, these findings document that vitamin E deficiency is responsible of mitochondrial morphometric alterations in adult rats. Structurally deteriorated mitochondria are reported to play a role in producing increased amounts of free radicals, which can facilitate the accumulation of ceroid pigment.     

Development of a Microarray-Based Tool To Characterize Vaginal Bacterial Fluctuations and Application to a Novel Antibiotic Treatment for Bacterial Vaginosis

The healthy vaginal microbiota is generally dominated by lactobacilli that confer antimicrobial protection and play a crucial role in health. Bacterial vaginosis (BV) is the most prevalent lower genital tract infection in women in reproductive age and is characterized by a shift in the relative abundances of Lactobacillus spp. to a greater abundance of strictly anaerobic bacteria. In this study, we designed a new phylogenetic microarray-based tool (VaginArray) that includes 17 probe sets specific for the most representative bacterial groups of the human vaginal ecosystem. This tool was implemented using the ligase detection reaction-universal array (LDR-UA) approach. The entire probe set properly recognized the specific targets and showed an overall sensitivity of 6 to 12 ng per probe. The VaginArray was applied to assess the efficacy of rifaximin vaginal tablets for the treatment of BV, analyzing the vaginal bacterial communities of 22 BV-affected women treated with rifaximin vaginal tablets at a dosage of 25 mg/day for 5 days. Our results showed the ability of rifaximin to reduce the growth of various BV-related bacteria (Atopobium vaginae, Prevotella, Megasphaera, Mobiluncus, and Sneathia spp.), with the highest antibiotic susceptibility for A. vaginae and Sneathia spp. Moreover, we observed an increase of Lactobacillus crispatus levels in the subset of women who maintained remission after 1 month of therapy, opening new perspectives for the treatment of BV.     

Design of donecopride,a dual serotonin subtype 4 receptor agonist/acetylcholinesterase inhibitor with potential interest for Alzheimer's disease treatment

RS67333 is a partial serotonin subtype 4 receptor (5-HT₄R) agonist that has been widely studied for its procognitive effect. More recently, it has been shown that its ability to promote the nonamyloidogenic cleavage of the precursor of the neurotoxic amyloid-β peptide leads to the secretion of the neurotrophic protein sAPPα. This effect has generated great interest in RS67333 as a potential treatment for Alzheimer’s disease (AD). We show herein that RS67333 is also a submicromolar acetylcholinesterase (AChE) inhibitor and therefore, could contribute, through this effect, to the restoration of the cholinergic neurotransmission that becomes altered in AD. We planned to pharmacomodulate RS67333 to enhance its AChE inhibitory activity to take advantage of this pleiotropic pharmacological profile in the design of a novel multitarget-directed ligand that is able to exert not only a symptomatic but also, a disease-modifying effect against AD. These efforts allowed us to select donecopride as a valuable dual (h)5-HT₄R partial agonist (K_i = 10.4 nM; 48.3% of control agonist response)/(h)AChEI (IC₅₀ = 16 nM) that further promotes sAPPα release (EC₅₀ = 11.3 nM). Donecopride, as a druggable lead, was assessed for its in vivo procognitive effects (0.1, 0.3, 1, and 3 mg/kg) with an improvement of memory performances observed at 0.3 and 1 mg/kg on the object recognition test. On the basis of these in vitro and in vivo activities, donecopride seems to be a promising drug candidate for AD treatment.Among the large family of serotonin receptors (5-HTR), some of them, such as the subtype 4 (5-HT₄R), are of particular interest in improving memory performance and therefore, decreasing memory deficits, such as those that occur in Alzheimer''s disease (AD). In the CNS, they are located in structures that are primarily involved in cognitive functions, like the olfactory tubercles, basal ganglia, septum, substantia nigra, superior colliculi, hippocampus, and cortex. Several compounds act as agonists to 5-HT₄R (BIMU1, BIMU8, RS17017, SL65.0155, VRX-03011, prucalopride, RS67333, and RS67506). One of the most affine (pK_i = 7.88) and selective vs. other receptors is RS67333, which acts as a partial agonist (1). With respect to the potential therapeutic modulation of 5-HT₄R with RS67333 and excluding its putative antidepressant-like activity (2–4), most studies focused on the promnesic or antiamnesic actions of this compound. These effects on cognitive functions that concern learning and memory are probably, in part, because of the fact that the pharmacological stimulation of these receptors increases the release of ACh in the hippocampus and cortex, and it also increases serotonin, dopamine, and GABA release (5–13). Concerning the selective aspects of memory functions, RS67333 has been shown to improve object recognition in adult (14, 15) and aged animals (16, 17) and place recognition (10) in rodents. It also increases spatial learning on the Morris water maze task in rodents, and it even reverses the deleterious effect of atropine (18) or scopolamine during this same task.Based on the structural analogy existing between RS67333 and donepezil (Fig. 1), we postulated that RS67333 could improve learning and memory by not only activating 5-HT₄R but also, inhibiting acetylcholinesterase (AChE) activity. Indeed, several early studies reported that the inhibition of AChE improved cognition and that this effect is the main reason for the initial use of donepezil, galantamine, and rivastigmine as cognitive enhancers in AD (19, 20). AChE inhibition has also been reported to improve performances in healthy rodents or animal models of memory deficiency (21). The hypothesis of an involvement of AChE inhibition by RS67333 in memory improvement is an issue that has never been tested.Open in a separate windowFig. 1.RS67333 and donepezil are chemically close.Furthermore, such a pharmacological profile could be also exploited to lead to pleiotropic compounds that are theoretically useful in AD treatment. Indeed, today, it is well-established that 5-HT₄R activation not only favors ACh release but also, is involved in the nonamyloidogenic cleavage of amyloid precursor protein (APP) in the neurotrophic sAPPα fragment, with secretion that is detrimental to amyloid-β peptide (Aβ) production (22–24). However, inhibiting the catalytic activity of AChE is widely used to restore cholinergic neurotransmission in AD, and interacting with the peripheral anionic site (PAS) of this enzyme could also reduce amyloid aggregation, for which AChE would be responsible (25). These activities seem to be synergistic when they are associated in an AD animal model, which we have recently shown in mice (26). A second pharmacological approach based on the fact that a single compound may be able to hit multiple targets is now emerging. This concept, called multitarget-directed ligands (MTDLs) (27, 28), would have inherent advantages over a combination of drugs called multiple medication therapy. It would specially obviate the problems linked to the complexity of the pharmacokinetic profile of the combined drugs and the risk of drug–drug interactions. Moreover, MTDL could also alleviate compliance difficulties associated with multiple medication therapy. It has also been shown that MTDLs generally show a higher synergistic effect than that observed with a combination of drugs. Numerous examples of MTDL against AD have been recently described (27). Most of them associate an AChE inhibitory effect with another activity hitting another molecular target of AD, such as antioxidant effect, monoamine oxidase inhibition, calcium channel blocking effect, metal chelating activity, etc. However, no MTDLs associating an inhibition of AChE and 5-HT₄R agonist effect have been hitherto described. Among the different ways to synthesize such MTDLs, one of them is to merge the frameworks of two selective starting compounds, each one exerting an activity toward a sole target (28). This goal is even more easily reached if the starting compounds are structurally close, and it is the reason why we considered the structural analogy between donepezil and RS67333 as a good starting point to design MTDLs displaying both activities (AChE inhibition and 5-HT₄R agonist activities). We will, thus, provide proof of this concept with the synthesis and biological evaluation of MR31147 (donecopride) as conceived from the pharmacomodulation of RS67333.     

Platelet-rich plasma injections for the treatment of refractory Achilles tendinopathy: results at 4 years

Background

Chronic Achilles tendinopathy is responsible for a severe reduction in physical performance and persistent pain. There is currently a number of therapeutic options and the local administration of growth factors is an emerging treatment strategy. In particular, platelet-rich plasma (PRP) is a widely used way to provide a local regenerative stimulus for tendon healing. The aim of this study was to document the mid-term results obtained after treating recalcitrant Achilles tendinopathy with injections of high concentrate, leucocyte-rich PRP.

Materials and methods

Twenty-seven patients (mean age: 44.6 years; 22 men and 5 women) affected by chronic mid-portion Achilles tendinopathy (7 bilateral, for a total of 34 tendons), refractory to previous treatments, were enrolled. Patients were treated with three ultrasound-guided intra-tendinous injections of PRP at 2-week intervals. Patients were prospectively evaluated at baseline, and then at 2, 6, and up to a mean of 54.1 months of follow-up (minimum 30 months), using the following tools: Blanzina, VISA-A, EQ-VAS for general health, and Tegner scores.

Results

The VISA-A score showed a significant improvement: the baseline score of 49.9±18.1 increased to 62.9±19.8 at 2 months (p=0.002), with a further improvement at 6 months (84.3±17.1, p<0.0005), and stable results at 4.5 years (90.0±13.9). The EQ-VAS score also showed a similar positive trend. An evaluation of the activity level confirmed these findings, showing a significant improvement in the Tegner score over time (p=0.017 for the final evaluation). The longer duration of symptoms before treatment was associated with a slower return to sport (p=0.041).

Discussion

PRP injections produced good overall results for the treatment of chronic recalcitrant Achilles tendinopathy with a stable outcome up to a medium-term follow-up. Longer symptom duration was related with a more difficult return to sporting activity.