Plasmodium falciparum Drug Resistance Genes pfmdr1 and pfcrt In Vivo Co-Expression During Artemether-Lumefantrine Therapy.

Background: Artemisinin-based combination therapies (ACTs) are the global mainstay treatment of uncomplicated Plasmodium falciparum infections. PfMDR1 and PfCRT are two transmembrane transporters, associated with sensitivity to several antimalarials, found in the parasite food vacuole. Herein, we explore if their relatedness extends to overlapping patterns of gene transcriptional activity before and during ACT administration. Methods: In a clinical trial performed in Tanzania, we explored the pfmdr1 and pfcrt transcription levels from 48 patients with uncomplicated P. falciparum malaria infections who underwent treatment with artemether-lumefantrine (AL). Samples analyzed were collected before treatment initiation and during the first 24 h of treatment. The frequency of PfMDR1 N86Y and PfCRT K76T was determined through PCR-RFLP or direct amplicon sequencing. Gene expression was analyzed by real-time quantitative PCR. Results: A wide range of pre-treatment expression levels was observed for both genes, approximately 10-fold for pfcrt and 50-fold for pfmdr1. In addition, a significant positive correlation demonstrates pfmdr1 and pfcrt co-expression. After AL treatment initiation, pfmdr1 and pfcrt maintained the positive co-expression correlation, with mild downregulation throughout the 24 h post-treatment. Additionally, a trend was observed for PfMDR1 N86 alleles and higher expression before treatment initiation. Conclusion: pfmdr1 and pfcrt showed significant co-expression patterns in vivo, which were generally maintained during ACT treatment. This observation points to relevant related roles in the normal parasite physiology, which seem essential to be maintained when the parasite is exposed to drug stress. In addition, keeping the simultaneous expression of both transporters might be advantageous for responding to the drug action.

Global plant trait relationships extend to the climatic extremes of the tundra biome.

The majority of variation in six traits critical to the growth, survival and reproduction of plant species is thought to be organised along just two dimensions, corresponding to strategies of plant size and resource acquisition. However, it is unknown whether global plant trait relationships extend to climatic extremes, and if these interspecific relationships are confounded by trait variation within species. We test whether trait relationships extend to the cold extremes of life on Earth using the largest database of tundra plant traits yet compiled. We show that tundra plants demonstrate remarkably similar resource economic traits, but not size traits, compared to global distributions, and exhibit the same two dimensions of trait variation. Three quarters of trait variation occurs among species, mirroring global estimates of interspecific trait variation. Plant trait relationships are thus generalizable to the edge of global trait-space, informing prediction of plant community change in a warming world.

Traditional plant functional groups explain variation in economic but not size-related traits across the tundra biome.

AIM: Plant functional groups are widely used in community ecology and earth system modelling to describe trait variation within and across plant communities. However, this approach rests on the assumption that functional groups explain a large proportion of trait variation among species. We test whether four commonly used plant functional groups represent variation in six ecologically important plant traits. LOCATION: Tundra biome. TIME PERIOD: Data collected between 1964 and 2016. MAJOR TAXA STUDIED: 295 tundra vascular plant species. METHODS: We compiled a database of six plant traits (plant height, leaf area, specific leaf area, leaf dry matter content, leaf nitrogen, seed mass) for tundra species. We examined the variation in species-level trait expression explained by four traditional functional groups (evergreen shrubs, deciduous shrubs, graminoids, forbs), and whether variation explained was dependent upon the traits included in analysis. We further compared the explanatory power and species composition of functional groups to alternative classifications generated using post hoc clustering of species-level traits. RESULTS: Traditional functional groups explained significant differences in trait expression, particularly amongst traits associated with resource economics, which were consistent across sites and at the biome scale. However, functional groups explained 19% of overall trait variation and poorly represented differences in traits associated with plant size. Post hoc classification of species did not correspond well with traditional functional groups, and explained twice as much variation in species-level trait expression. MAIN CONCLUSIONS: Traditional functional groups only coarsely represent variation in well-measured traits within tundra plant communities, and better explain resource economic traits than size-related traits. We recommend caution when using functional group approaches to predict tundra vegetation change, or ecosystem functions relating to plant size, such as albedo or carbon storage. We argue that alternative classifications or direct use of specific plant traits could provide new insights for ecological prediction and modelling.

Plasmodium falciparum K13 expression associated with parasite clearance during artemisinin-based combination therapy.

BACKGROUND: Delayed parasite clearance and, consequently, reduced efficacy of artemisinin-based combination therapies have been linked with Plasmodium falciparum K13 gene SNPs in Southeast Asia. In Africa, significantly prolonged clearance has not yet been observed and the presently restricted variation in parasite clearance cannot be explained by K13 polymorphisms. OBJECTIVES: Our aim was to study the in vivo pfK13 transcriptional response in patients treated with artemether-lumefantrine and explore whether the pfk13 transcripts can explain the patients' parasite clearance outcomes. PATIENTS AND METHODS: A total of 47 Tanzanian children with microscopically confirmed uncomplicated P. falciparum malaria were hospitalized and received artemether-lumefantrine treatment (clinical trial ID: NCT00336375). RNA was extracted from venous blood samples collected before treatment initiation and at five more timepoints after treatment. cDNA was synthesized and pfk13 transcripts measured by real-time PCR. RESULTS: A wide range of pfk13 transcript variation was observed throughout all timepoints after artemether-lumefantrine treatment. Taking parasite clearance data together with the pfk13 transcripts profile, we observed a negative correlation inferring that pfk13 down-regulation is associated with longer parasite clearance time. CONCLUSIONS: The findings suggest that a reduced PfK13 transcriptional response may represent a first step towards artemisinin tolerance/resistance.

Avaliação psicométrica em português do indicador de dor crônica de Helsinki em cães com sinais crônicos de osteoartrite / Psychometric testing in Portuguese of Helsinki's chronic pain index for dogs with chronic signs of osteoarthritis

Objetivou-se determinar a validade, a confiabilidade e a sensibilidade do indicador de dor crônica de Helsinki (IDCH) em português, além de estabelecer correlação entre a plataforma de pressão em cães com displasia coxofemoral (DCF). O questionário foi preenchido por tutores de 40 animais com DCF e por 16 tutores de cães saudáveis. Os animais com DCF foram tratados com carprofeno 4,4mg/kg (GT= 21) ou placebo (GP= 19), administrados uma vez ao dia, durante quatro semanas. A avaliação consistiu no preenchimento do questionário (IDCH), na escala analógica visual de dor (EAVdor) e na escala analógica visual de locomoção (EAVloc) com duas semanas (A1) e imediatamente antes do tratamento (A2), duas (S2) e quatro (S4) semanas de tratamento e após duas semanas do término do tratamento (S6). A consistência interna dos dados foi considerada excelente (alfa de Cronbach = 0,89). Houve correlação moderada entre o IDCH e a EAVdor e boa entre o IDCH e a EAVloc, porém não houve diferença entre o grupo tratado com carprofeno e placebo no IDCH. Não houve correlação entre os dados da plataforma de pressão e o IDCH. O questionário apresentou validade de constructo, critério e fidedignidade e pode-se empregá-lo em países de língua portuguesa em cães com osteoartrite.(AU)

We aimed to determine validity, reliability, and sensitivity of Helsinki's chronic pain index (HCPI) and stablish a correlation between HCPI in dogs with hip dysplasia (HD) using pressure sensitive walkway. Forty-owners of dogs with HD and 16 owners of health dogs filled a questionnaire. Dogs with HD were treated with carprofen 4.4mg/kg (GT n= 21) or with placebo (GP n= 19), both were administered once a day for 4 weeks. Evaluation was performed by the owners using the questionnaire (HCPI), the Visual Analogue Scale for pain (VASpain) and the VAS for locomotion (VASloc). The evaluation was performed 2 weeks before the treatment began (A1), immediately after treatment (A2), two (S2), four (S4) and two weeks after the end of treatment (S6) and the lameness was evaluated by pressure sensitive walkway. The internal consistency of the data was considered excellent (Cronbach´α coefficient= 0.89). There was a moderate correlation between the HCPI and VASpain. For VASloc the correlation was good. However, there was no difference between treatments, indicating low sensibility. No correlation was observed between pressure sensitive walkway and HCPI. We concluded that the questionnaire has construct and criterion validity, reliability and can be applied in dogs with osteoarthritis in Portuguese-speaking countries.(AU)

From high to low malaria transmission in Zanzibar-challenges and opportunities to achieve elimination.

BACKGROUND: Substantial global progress in the control of malaria in recent years has led to increased commitment to its potential elimination. Whether this is possible in high transmission areas of sub-Saharan Africa remains unclear. Zanzibar represents a unique case study of such attempt, where modern tools and strategies for malaria treatment and vector control have been deployed since 2003. METHODS: We have studied temporal trends of comprehensive malariometric indices in two districts with over 100,000 inhabitants each. The analyses included triangulation of data from annual community-based cross-sectional surveys, health management information systems, vital registry and entomological sentinel surveys. RESULTS: The interventions, with sustained high-community uptake, were temporally associated with a major malaria decline, most pronounced between 2004 and 2007 and followed by a sustained state of low transmission. In 2015, the Plasmodium falciparum community prevalence of 0.43% (95% CI 0.23-0.73) by microscopy or rapid diagnostic test represented 96% reduction compared with that in 2003. The P. falciparum and P. malariae prevalence by PCR was 1.8% (95% CI 1.3-2.3), and the annual P. falciparum incidence was estimated to 8 infections including 2.8 clinical episodes per 1000 inhabitants. The total parasite load decreased over 1000-fold (99.9%) between 2003 and 2015. The incidence of symptomatic malaria at health facilities decreased by 94% with a trend towards relatively higher incidence in age groups > 5 years, a more pronounced seasonality and with reported travel history to/from Tanzania mainland as a higher risk factor. All-cause mortality among children < 5 years decreased by 72% between 2002 and 2007 mainly following the introduction of artemisinin-based combination therapies whereas the main reduction in malaria incidence followed upon the vector control interventions from 2006. Human biting rates decreased by 98% with a major shift towards outdoor biting by Anopheles arabiensis. CONCLUSIONS: Zanzibar provides new evidence of the feasibility of reaching uniquely significant and sustainable malaria reduction (pre-elimination) in a previously high endemic region in sub-Saharan Africa. The data highlight constraints of optimistic prognostic modelling studies. New challenges, mainly with outdoor transmission, a large asymptomatic parasite reservoir and imported infections, require novel tools and reoriented strategies to prevent a rebound effect and achieve elimination.

Accuracy of Pre- and Postcontrast, 3 T Indirect MR Arthrography Compared with Wrist Arthroscopy in the Diagnosis of Wrist Ligament Injuries.

Background Magnetic resonance (MR) is the most important imaging technique to assess intra-articular pathology of the wrist. Among various MR imaging protocols, the diagnostic performance of indirect MR arthrography needs further investigation. Purpose The purpose of this study was to assess the diagnostic performance of pre- and postcontrast, 3 T indirect MR arthrography in the diagnosis of scapholunate intrinsic ligament (SLIL) and triangular fibrocartilage complex (TFCC) injuries, using wrist arthroscopy as reference standard. Patients and Methods We retrospectively evaluated consecutive patients with suspected SLIL or TFCC injury, who had indirect MR arthrography done before arthroscopy. Images were assessed independently by two senior radiologists. Results Arthroscopy of the 53 wrists demonstrated 16 Geissler stages II and III partial tears and 6 stage IV total SLIL ruptures. Central perforation of the TFCC was found in 24 wrists, and 12 wrists had an ulnar class 1B lesion. To detect any SLIL tear, accuracy was higher for the two observers using postcontrast indirect MR arthrography (0.77 and 0.72) than for precontrast MR imaging (0.60 and 0.60). No difference was found for total SLIL ruptures "0.85 and 0.89" versus "0.85 and 0.89." To diagnose class 1B TFCC injuries, accuracy was higher using postcontrast indirect MR arthrography (0.85 and 0.75) than for precontrast MR imaging (0.70 and 0.72). No difference in accuracy was demonstrated for TFCC central tears "0.75 and 0.75" versus "0.70 and 0.77." Conclusion Postcontrast images at 3 T indirect MR arthrography, compared with precontrast images, have an improved diagnostic performance for the overall detection of SLIL injuries and as well as class 1B TFCC tears. Level of Evidence This is a Level II, diagnostic study.

Syringomyelia and Craniocervical Junction Abnormalities in Chihuahuas.

BACKGROUND: Chiari-like malformation (CM) and syringomyelia (SM) are widely reported in Cavalier King Charles Spaniels and Griffon Bruxellois dogs. Increasing evidence indicates that CM and SM also occur in other small and toy breed dogs, such as Chihuahuas. OBJECTIVES: To describe the presence of SM and craniocervical junction (CCJ) abnormalities in Chihuahuas and to evaluate the possible association of CCJ abnormalities with SM. To describe CM/SM-related clinical signs and neurologic deficits and to investigate the association of CM/SM-related clinical signs with signalment, SM, or CCJ abnormalities. ANIMALS: Fifty-three client-owned Chihuahuas. METHODS: Prospective study. Questionnaire analyses and physical and neurologic examinations were obtained before magnetic resonance and computed tomography imaging. Images were evaluated for the presence of SM, CM, and atlantooccipital overlapping. Additionally, medullary kinking, dorsal spinal cord compression, and their sum indices were calculated. RESULTS: Scratching was the most common CM/SM-related clinical sign and decreased postural reaction the most common neurologic deficit in 73 and 87% of dogs, respectively. Chiari-like malformation and SM were present in 100 and 38% of dogs, respectively. Syringomyelia was associated with the presence of CM/SM-related clinical signs (P = 0.034), and medullary kinking and sum indices were higher in dogs with clinical signs (P = 0.016 and P = 0.007, respectively). CONCLUSIONS AND CLINICAL IMPORTANCE: Syringomyelia and CCJ abnormalities are prevalent in Chihuahuas. Syringomyelia was an important factor for the presence of CM/SM-related clinical signs, but many dogs suffered from similar clinical signs without being affected by SM, highlighting the clinical importance of CCJ abnormalities in Chihuahuas.

Behavioral Abnormalities in Lagotto Romagnolo Dogs with a History of Benign Familial Juvenile Epilepsy: A Long-Term Follow-Up Study.

BACKGROUND: Lagotto Romagnolo (LR) dogs with benign juvenile epilepsy syndrome often experience spontaneous remission of seizures. The long-term outcome in these dogs currently is unknown. In humans, behavioral and psychiatric comorbidities have been reported in pediatric and adult-onset epilepsies. HYPOTHESIS/OBJECTIVES: The objectives of this study were to investigate possible neurobehavioral comorbidities in LR with a history of benign familial juvenile epilepsy (BFJE) and to assess the occurrence of seizures after the remission of seizures in puppyhood. ANIMALS: A total of 25 LR with a history of BFJE and 91 control dogs of the same breed. METHODS: Owners of the LR dogs in the BFJE and control groups completed an online questionnaire about each dog's activity, impulsivity, and inattention. Principal component analysis (PCA) served to extract behavioral factors from the data. We then compared the scores of these factors between the 2 groups in a retrospective case-control study. We also interviewed all dog owners in the BFJE group by telephone to inquire specifically about possible seizures or other neurological problems after remission of seizures as a puppy. RESULTS: Lagotto Romagnolo dogs with BFJE showed significantly higher scores on the factors Inattention and Excitability/Impulsivity than did the control group (P = .003; P = .021, respectively). Only 1 of the 25 BFJE LR exhibited seizures after remission of epilepsy in puppyhood. CONCLUSIONS AND CLINICAL IMPORTANCE: Although the long-term seizure outcome in BFJE LR seems to be good, the dogs exhibit behavioral abnormalities resembling attention deficit hyperactivity disorder (ADHD) in humans, thus suggesting neurobehavioral comorbidities with epilepsy.

Enhanced early sensory outcome after nerve repair as a result of immediate post-operative re-learning: a randomized controlled trial.

We assessed the use of guided plasticity training to improve the outcome in the first 6 months after nerve repair. In a multicentre randomized controlled trial, 37 adults with median or ulnar nerve repair at the distal forearm were randomized to intervention, starting the first week after surgery with sensory and motor re-learning using mirror visual feedback and observation of touch, or to a control group with re-learning starting when reinnervation could be detected. The primary outcome at 3 and 6 months post-operatively was discriminative touch (shape texture identification test, part of the Rosen score). At 6 months, discriminative touch was significantly better in the early intervention group. Improvement of discriminative touch between 3 and 6 months was also significantly greater in that group. There were no significant differences in motor function, pain or in the total score. We conclude that early re-learning using guided plasticity may have a potential to improve the outcomes after nerve repair. LEVEL OF EVIDENCE II.

Intra-articular botulinum toxin A for the treatment of osteoarthritic joint pain in dogs: a randomized, double-blinded, placebo-controlled clinical trial.

The aim of this study was to investigate the efficacy of intra-articular (IA) botulinum toxin A (IA BoNT A) for the treatment of osteoarthritic joint pain in dogs. The study was a placebo-controlled, randomized, double-blinded clinical trial with parallel group design and 12-week follow-up. Thirty-six dogs with chronic lameness due to stifle, hip or elbow osteoarthritis were randomized to receive an IA injection of 30IU of BoNT A or placebo. The main outcome variables were vertical impulse (VI) and peak vertical force (PVF) measured with a force platform, and Helsinki chronic pain index (HCPI). Subjective pain score and the need for rescue analgesics were secondary variables. The response to treatment was assessed as the change from baseline to each examination week. The variables were analyzed by ANOVA with repeated measurements and results were considered statistically significant if P ⩽ 0.05. The improvement from baseline to 12 weeks after baseline was statistically significant in VI, PVF and HCPI in the treatment group (P=0.001, P=0.054 and P=0.053, respectively). Additionally, there were statistically significant improvements in VI in the treatment group at 2, 4 and 8 weeks after baseline (P=0.037, P=0.016 and P=0.016, respectively). The difference between groups in improvement in VI was statistically significant at 12weeks after baseline (P=0.005). There was no significant change in the subjective pain score or in the requirement for rescue analgesics in either group. No major adverse events thought to be related to trial protocol were detected. These results suggest that IA BoNT A has some efficacy in reducing osteoarthritic pain in dogs.

Topiramate as an add-on antiepileptic drug in treating refractory canine idiopathic epilepsy.

OBJECTIVES: To evaluate the efficacy and safety of topiramate as an add-on therapy in dogs with refractory idiopathic epilepsy. METHOD: Prospective, open label, non-comparative clinical trial of topiramate in dogs with idiopathic epilepsy and poor seizure control despite therapeutic serum concentrations of phenobarbital and potassium bromide. The efficacy of topiramate was evaluated by comparing seizure and seizure day frequencies during a retrospective 2-month period with a prospective short-term follow-up of 6 months. An additional long-term follow-up period ranging from 3 to 9 months was conducted on dogs that responded to topiramate therapy during the short-term follow-up. RESULTS: Ten dogs were included. Five (50%) responded to topiramate therapy during the short-term follow-up showing a significant (P=0·04) decrease of 66% in seizure frequency. Three of the five dogs remained responders during the long-term follow-up. Weight loss, sedation and ataxia were the most common adverse effects of topiramate therapy, but in dogs with moderate sedation or ataxia, signs subsided in a few weeks to few months to mild sedation or ataxia. CLINICAL SIGNIFICANCE: Topiramate may be effective as an add-on medication in treating canine idiopathic epilepsy. Apart from sedation and ataxia reported in some of the dogs, topiramate was well-tolerated.

Functional outcome thirty years after median and ulnar nerve repair in childhood and adolescence.

BACKGROUND: Age at injury is believed to be a factor that strongly influences functional outcome after nerve injury. However, there have been few long-term evaluations of the results of nerve repair and reconstruction in children. Our aim was to evaluate the long-term functional outcome of nerve repair or reconstruction at the forearm level in patients with a complete median and/or ulnar nerve injury at a young age. METHODS: Forty-five patients were assessed at a median of thirty-one years after a complete median and/or ulnar nerve injury in the forearm. The outcome was classified with a total score (the Rosén score), a standardized outcome instrument consisting of three separate domains for sensory and motor function as well as pain/discomfort. In addition, the DASH (Disabilities of the Arm, Shoulder and Hand) score, sensitivity to cold, and locognosia were assessed specifically, together with the patient's estimation of the overall outcome and impact on his or her education, work, and leisure activities. Comparisons were made between injuries that occurred in childhood (less than twelve years of age) and those that were sustained in adolescence (twelve to twenty years of age), and according to the nerve(s) that was injured (median nerve, ulnar nerve, or both). RESULTS: Functional recovery, expressed as the total outcome score, the sensory domain of that score, and the patient's subjective estimation of outcome, was significantly better after injuries sustained in childhood than after those that occurred in adolescence (87% and 67% of complete recovery, respectively; p < 0.001). No significant differences in recovery were seen between median and ulnar nerve injuries, or even when both nerves were injured. Motor function was close to normal, and cold sensitivity was not a problem in either age group. The median DASH scores were within normal limits and did not differ between the groups. Patients who sustained the injury in adolescence indicated that the nerve injury had a significantly higher effect on their profession, education, and leisure activities. CONCLUSIONS: At a median of thirty-one years after a median or ulnar nerve repair at the level of the forearm, nerve function is significantly better in those injured in childhood than in those injured in adolescence, with almost full sensory and motor recovery in individuals injured in childhood.

Vibration thresholds are increased at low frequencies in the sole of the foot in diabetes-a novel multi-frequency approach.

AIMS: To evaluate multi-frequency tactilometry as a method to measure vibrotactile sense in the sole of the foot in subjects with diabetes. METHODS: Vibration thresholds were investigated at five frequencies (8, 16, 32, 64 and 125 Hz) at three sites (first and fifth metatarsal heads and heel) in the sole of the foot in subjects with Type 1 and Type 2 diabetes (n = 37). Thresholds were compared with healthy, age- and gender-matched subjects (n = 37) and related to glycaemic levels, subjective estimation of sensation in the feet and to perception of touch. RESULTS: Vibration thresholds were significantly higher in subjects with diabetes compared with healthy subjects at low frequencies (8, 16 and 32 Hz) at all measured sites, and also at 64 Hz for the metatarsal heads. Perception of touch and subjective estimation of sensation were significantly impaired in subjects with diabetes. Glycaemic levels, which were higher in subjects with diabetes, did not correlate with vibration thresholds at 32 Hz (most sensitive to Meissner's corpuscles) or with touch thresholds in subjects with diabetes. Vibration thresholds at 32 Hz correlated significantly with perception of touch (rho = 0.45-0.65; P < 0.01) and with subjective sensation (rho = -0.38 to -0.52; P < 0.001) in subjects with diabetes. Perception of touch and subjective estimation of sensation did also correlate (rho = -0.51 to -0.80; P < 0.002). CONCLUSIONS: Tactilometry is effective in detecting neuropathy in the sole of the foot at low frequencies of mainly 8-32 Hz, indicating that at least Meissner's corpuscles, or their related large nerve fibres, are affected by diabetes.

Use of bathroom scales in measuring asymmetry of hindlimb static weight bearing in dogs with osteoarthritis.

OBJECTIVE: The study assessed the use and reliability of bathroom scales as an objective measurement tool, and setting a normal variance of static weight bearing between hindlimbs. METHODS: Two groups of dogs were tested: a healthy control group (n = 21) and a group (n = 43) of dogs with confirmed osteoarthritis in at least one stifle joint, with or without hip joint osteoarthritis. Static weight bearing was evaluated manually and measured with two bathroom scales. An orthopaedic examination was done and dynamic weight bearing was measured using a force platform. Radiographs were taken to confirm the presence of osteoarthritis, and dogs were divided into groups of severe and non-severe osteo- arthritic changes. Reliability by repeatability was tested using analysis of variance, and the congruity between static weight bearing and other evaluation methods with Kappa statistics and proportion of agreement. RESULTS: The difference between the hindlimbs proportional to the body weight in control dogs was 3.3% (± 2.7%). The repeatability of measuring static weight bearing in the hindlimbs of osteoarthritic dogs with bathroom scales was 81% with osteoarthritic limbs, and 70% for unaffected limbs. The sensitivity of static weight bearing measurements using bathroom scales was 39% and specificity 85%. CLINICAL SIGNIFICANCE: Bathroom scales are a reliable, simple, and cost-effective objective method for measuring static weight bearing and can be used as an outcome measure when rehabilitating dogs with osteoarthritic changes in the hindlimbs.

Canine total knee replacement performed due to osteoarthritis subsequent to distal femur fracture osteosynthesis: two-year objective outcome.

A 27-kg German Shorthaired Pointer was referred for evaluation due to the complaint of left pelvic limb lameness and signs of pain in the left stifle joint. Radiographs revealed signs of a healed supracondylar femoral fracture that had been previously repaired at another hospital with an intramedullary pin and two cross pins. In addition, there were signs of severe osteoarthritis (OA). The OA had been managed medically with administration of carprofen and nutraceuticals for nine months without any improvement. Left total knee replacement (TKR) surgery was performed to alleviate signs of pain. The patient was assessed preoperatively and at six months, one year, and two years after surgery using radiology, force platform analysis of gait, thigh circumference measures, goniometry, and lameness evaluation. Following surgery, the dog resumed normal activity without any signs of pain and a good quality of life at 3.5 months. Force plate analysis found that peak vertical force on the TKR limb was 85.7% of the normal contralateral limb after two years. Total knee replacement was a successful treatment to manage knee OA associated with a healed distal femoral fracture and internal fixation in this dog.

Cytochrome 1A1 and 1B1 gene diversity in the Zanzibar islands.

Amodiaquine (AQ) is a 4-aminoquinoline widely used in the treatment of malaria as part of the artemisinin combination therapy (ACT). AQ is metabolised towards its main metabolite desethylamodiaquine mainly by cytochrome P450 2C8 (CYP2C8). CYP1A1 and CYP1B1 play a minor role in the metabolism but they seem to be significantly involved in the formation of the short-lived quinine-imine. To complete the genetic variation picture of the main genes involved in AQ metabolism in the Zanzibar population, previously characterised for CYP2C8, we analysed in this study CYP1A1 and CYP1B1 main genetic polymorphisms. The results obtained show a low frequency of the CYP1A1*2B/C allele (2.4%) and a high frequency of CYP1B1*6 (approximately 42%) followed by CYP1B1*2 (approximately 27%) in Zanzibar islands. Genotype data for CYP1A1 and CYP1B1 show a low incidence of fast metabolisers, revealing a relatively safe genetic background in Zanzibar's population regarding the appearance of adverse effects.

pfmdr1 amplification is related to increased Plasmodium falciparum in vitro sensitivity to the bisquinoline piperaquine.

The 4-aminoquinoline bisquinoline piperaquine is an important partner drug in one of the presently recommended artemisinin combination therapies. Recent clinical trials have confirmed its high efficacy in combination with dihydroartemisinin. Resistance to piperaquine alone has, however, been documented. Amplification in copy number of the Plasmodium falciparum multidrug resistance locus on chromosome 5, containing the pfmdr1 gene, has been shown to confer resistance to structurally unrelated antimalarials. Through the determination of the 50% inhibitory concentrations (IC(50)s) and IC(90)s for piperaquine and chloroquine in a set of 46 adapted P. falciparum cultures originating from the Thai-Burmese border, we have characterized the regions around the pfmdr1 gene and identified a significant association between the presence of pfmdr1 duplications and enhanced sensitivity to piperaquine (P = 0.005 for IC(50) and P = 0.002 for IC(90)) and chloroquine, reaching statistical significance at IC(90)s (P = 0.026). These results substantiate the potential importance of pfmdr1 copy number amplifications in the efficacy of the combination therapy piperaquine-dihydroartemisinin. It supports the rational use of 4-aminoquinolines and artemisinin-based compounds, as they independently select for mutually incompatible combinations of mutations.

Cutaneous anaesthesia of the lower leg can improve sensibility in the diabetic foot. A double-blind, randomized clinical trial.

AIMS: Impaired sensory function in the sole of the foot in diabetic patients is a substantial problem caused by unknown mechanisms. Hand or foot sensibility can be improved by cutaneous anaesthesia of the forearm or lower leg, respectively, in healthy subjects. Hypothetically, cutaneous anaesthesia induces a silent area in the primary somatosensory cortex, allowing adjacent cortical areas to expand; thus, resulting in enhanced sensory processing. Our aim was to improve sensory function in the foot in Type 1 and Type 2 diabetic patients by application of an anaesthetic cream to the lower leg. METHODS: In a double-blind study, 37 patients with Type 1 or Type 2 diabetes were randomly assigned to cutaneous application of either an anaesthetic cream (EMLA) or a placebo cream to the skin of the lower leg for 1.5 h. Sensibility at five points of the sole of the foot was assessed before and after 1.5 and 24 h. Vibrotactile sense was also assessed. Primary outcome was change of touch threshold at the first metatarsal head from pretreatment to 1.5 h assessment. RESULTS: Anaesthetic cream on the lower leg resulted in a significant improvement of touch threshold at the first metatarsal head after 1.5 and 24 h. In addition, improvement of touch thresholds was also observed at the other four assessment sites, together with a decreased vibration threshold at 125 Hz. CONCLUSIONS: The findings of improved touch thresholds open up new possibilities in treatment of sensibility disturbances in the diabetic foot, using a simple and non-invasive method.

Pharmacogenomics of CYP2A6, CYP2B6, CYP2C19, CYP2D6, CYP3A4, CYP3A5 and MDR1 in Vietnam.

AIM: The aim of this study was to obtain pharmacogenetic data in a Vietnamese population on genes coding for proteins involved in the elimination of drugs currently used for the treatment of malaria and human immunodeficiency virus/acquired immunodeficiency syndrome. METHOD: The main polymorphisms on the cytochrome P450 (CYP) genes, CYP2A6, CYP2B6, CYP2C19, CYP2D6, CYP3A4 and CYP3A5, and the multi-drug resistance 1 gene (MDR1) were genotyped in 78 healthy Vietnamese subjects. Pharmacokinetic metrics were available for CYP2A6 (coumarin), CYP2C19 (mephenytoin), CYP2D6 (metoprolol) and CYP3As (midazolam), allowing correlations with the determined genotype. RESULTS: In the CYP2 family, we detected alleles CYP2A6*4 (12%) and *5 (15%); CYP2B6*4 (8%), *6 (27%); CYP2C19*2 (31%) and *3 (6%); CYP2D6*4, *5, *10 (1, 8 and 44%, respectively). In the CYP3A family, CYP3A4*1B was detected at a low frequency (2%), whereas CYP3A5 *3 was detected at a frequency of 67%. The MDR1 3435T allele was present with a prevalence of 40%. Allele proportions in our cohort were compared with those reported for other Asian populations. CYP2C19 genotypes were associated to the S-4'-OH-mephenytoin/S-mephenytoin ratio quantified in plasma 4 h after intake of 100 mg mephenytoin. While CYP2D6 genotypes were partially reflected by the alpha-OH-metroprolol/metoprolol ratio in plasma 4 h after dosing, no correlation existed between midazolam plasma concentrations 4 h post-dose and CYP3A genotypes. CONCLUSIONS: The Vietnamese subjects of our study cohort presented allele prevalences in drug-metabolising enzymes that were generally comparable with those reported in other Asian populations. Deviations were found for CYP2A6*4 compared to a Chinese population (12 vs. 5%, respectively; P = 0.023), CYP2A6*5 compared with a Korean population (15 vs. <1%, respectively; P < 0.0001), a Malaysian population (1%; P < 0.0001) and a Chinese population (1%; P < 0.0001); CYP2B6*6 compared with a Korean population (27 vs. 12%; P = 0.002) and a Japanese population (16%; P = 0.021). Pharmacokinetic metrics versus genotype analysis reinforces the view that the predictive value of certain globally common variants (e.g. CYP2D6 single nucleotide polymorphisms) should be evaluated in a population-specific manner.