Methods for comparing the diversity of samples of the T cell receptor repertoire

Analysis of T cell receptor (TCR) data has become a crucial element in many studies aimed at better understanding the evolution of the T cell repertoire and the role of TCR diversity in immune responses. In this paper we focus on comparing the diversity between samples of the TCR repertoire. We discuss some of the limitations and potential problems inherent in some of the more popular approaches to comparing samples of the TCR repertoire and we suggest alternate methods that both avoid these problems and enrich the analysis of TCR data. Examples from published studies of the CD8(+) T cell responses to the influenza A virus D(b)NP(366) and D(b)PA(224) epitopes in mice are used to demonstrate the implementation of these methods. One example involves a comparison between the central and effector memory T cell subsets, defined on the basis of CD62L expression, and the other examines changes in the TCR repertoire over time.     

The influence of smear layer in lateral channels filling

This in vitro study evaluated the ability of a warm gutta-percha obturation system Thermafil to fill lateral channels in presence/absence of smear layer. Forty single-rooted extracted human teeth were randomly divided into two groups for which different irrigation regimens were used: group A, 5 ml of 5% NaOCl + 2.5 ml of 3.6% H₂O₂; group B, 5 ml of 5% NaOCl 5% + 2.5 ml of 17% ethylenediamine tetraacetic acid. A conventional crown-down preparation technique was employed. Obturation was performed using epoxy resin-based cement (AH Plus) and a warm gutta-percha plastic carrier system (Thermafil). Specimens were cleared in methyl salicylate and analyzed under a stereomicroscope to evaluate the number, length, and diameter of lateral channels. Lateral channels were identified in both groups at medium and apical thirds. Additional samples were prepared for scanning electron microscopy inspection to confirm the presence of smear layer in group A, and the absence of smear layer in group B. All lateral channels resulted filled in both groups. No statistically significant differences regarding number, length, and diameter were observed between the two groups. Smear layer did not prevent the sealing of lateral channels.     

Cutaneous adverse reactions linked to targeted anticancer therapies bortezomib and lenalidomide for multiple myeloma: new drugs,old side effects

Context: Cutaneous toxicity is a frequent side effect of new anticancer targeted therapies. Skin reactions can severely impact the patient’s physical, psychological and social well-being and may sometimes lead to discontinuations either treatment dose reductions.

Objective: This study evaluates the impact of cutaneous adverse drug reactions (cADR) of the new therapies bortezomib and lenalidomide and presents a review of their skin side effects.

Materials and method: Type, frequency, severity, time of onset and management of cADR were collected and the medical records of all multiple myeloma patients receiving bortezomib or lenalidomide in the Hematology and Medical Oncology Institute of the University of Bologna, were analyzed.

Results: A total of 17 cADR occurred in 10 patients of 17 (58.8% of patients) treated with bortezomib: 5 rashes, 3 events of pruriginous rash, 1 purpuric rash, 2 records of mouth swelling, 1 stomatitis-mucositis, 3 cases of edema in the lower limbs, 1 patient referred pruritus and another telogen effluvium. Eight skin manifestations were due to lenalidomide in 7 patients of 25 treated (28%): 2 pruriginous rashes, 3 cases of edema, 2 records of pruritus, 1 case of stomatitis-mucositis. Three adverse events linked to bortezomib and 4 to lenalidomide forced to a complete withdrawal of the drug, while 3 reactions due to bortezomib mandated a dose reduction. Dermatological evaluation was performed only in 2 patients treated with bortezomib and 1 with lenalidomide.

Discussion: Evaluations of cADR due to bortezomib and lenalidomide were performed. There are no other reports focused on skin events in patients treated with the triple regimen velcade (bortezomib)-thalidomide-dexamethasone (VTD) up to date. Our study suggests that cutaneous toxicities, when researched by Dermatologists, are a side effect even more frequent than the reported data.

Limitations: As it is a single institute and retrospective study, ongoing cADR were rarely evaluated by dermatologists; thus, it is possible that cutaneous reactions (especially mild) may have been under reported by Hematologists and Oncologists in clinical records.

Conclusions: Even with the development of new drugs for cancer treatment, “old” cutaneous side effects may still be present, compromising patients’ quality of life. Physicians prescribing bortezomib and lenalidomide should monitor their patients for the spectrum of cADR, and they should involve dermatologists in consultations and management of these events. A multidisciplinar approach is necessary to oncologic patient in order to provide a tailored supportive clinical care.     

Selection criteria and long-term results of surgery in symptomatic rectocele

BACKGROUND: The aim of this study was to confirm some selection criteria for the transrectal repair of the anterior rectocele and to compare our surgical results with those reported in the literature. METHODS: From January 1992 to December 1999, 30 females (mean age 52.9 years, range 28-70 yrs) affected by anterior rectocele were prospectively evaluated with a standard questionnaire, clinical examination, proctosigmoidoscopy, colonic transit time, dynamic defecography, anal EMG, anal manometry. Then, they were submitted to transrectal repair of rectocele with anterior plication of the rectal muscular wall. Fourteen (46.6%) of them were also submitted to perineal levatorplasty. Patients were followed postoperatively (mean 25.7 months) with the same standard questionnaire, clinical examination, defecography, and manometry. Results were tested by Fisher's Exact text, Wilcoxon's test, and "t"-test. RESULTS: Rectal dyschezia, incomplete evacuation, digital help in defecating, mean stool frequency, and rectal bleeding significantly improved. After 3 months, 30% of patients had no complaints, 40% had only 1-2 episodes/month complaints, 13.3% had evacuation only using laxatives, and 16.6% were unchanged. Defecography showed a significant reduction of the rectocele in 70% of patients after 3 months. Manometric parameters were not significantly modified. Four (28.6%) out of 14 patients submitted to perineal levatorplasty complained of dyspareunia. CONCLUSIONS: Our surgical results were comparable with those reported in the literature, with more than 80% of successful outcome. Preoperative clinical data and defecography were confirmed to be basic parameters in selecting patients for surgery. Colonic transit time, anal EMG, and anorectal manometry demonstrated to be useful to recognize conditions as slow colonic transit time, peripheral denervation, and reduced voluntary contraction that could lead to a less satisfactory outcome after surgery, and might benefit with a postoperative perineal rehabilitation by biofeedback and anal electrostimulations. The perineal levatorplasty is not suitable in young females, due to the risk of dyspareunia.     

Cerebrotendinous xanthomatosis (van Bogaert-Scherer-Epstein disease): CT and MR findings.

PURPOSETo describe the CT and MR findings in the brain and spinal cord of patients with cerebrotendinous xanthomatosis and to seek possible correlations between clinical, biochemical (cholestanol levels), and neuroimaging findings.METHODSTen patients with well-defined clinical and biochemical diagnoses of cerebrotendinous xanthomatosis were examined. Brain CT was performed in eight cases. In all patients MR was obtained using spin-echo and gradient-echo sequences. In eight patients spine MR was also performed.RESULTSNeuroradiologic findings included diffuse cerebral and cerebellar atrophy. In half the cases, atrophy of the brain stem and corpus callosum was also found. In the majority of patients cerebellar bilateral focal lesions and mild white matter signal alterations were present. Spinal cord MR did not show signal abnormalities or atrophy.CONCLUSIONSWe found cranial alterations in patients with severe neurologic impairment, but there was no correlation with cholestanol plasma levels. No spinal cord abnormalities were present.     

Manometric study of the anal sphincter during haemorrhoidectomy using posterior perineal block

Minor anorectal diseases affect 4-5% of the adult western population. Operations are performed on an ambulatory or 24-hour-stay basis. The aim of our study was to assess the physiology of anal sphincter relaxation by anal manometry after posterior perineal block during haemorrhoidectomy. We recruited 15 patients with third and fourth degree hemorrhoids in a manometric study of the anal sphincter during haemorrhoidectomy with regional anaesthesia. The patients underwent anal manometry before and 15 minutes after the posterior perineal block to determine the resting and squeeze anal pressures. Differences were considered significant at p < 0.05. We observed mean reductions of 34.6% and 37.1% in resting and squeeze pressure values, respectively, after posterior perineal block (p < 0.005). Our manometric study demonstrated that anal sphincter relaxation after posterior perineal block correlates with a significant reduction in resting and squeeze pressures because the block anaesthesia not only the somatic, but also the sympathetic fibres. We believe that posterior perineal block allows the surgeon to perform radical haemorrhoidectomy in the overnight stay setting with optimal intra- and postoperative analgesia, safe sphincter relaxation, lower postoperative complications, and lower costs to the public health service.     

Indolizine derivativatives with biological activity III: 3-(3-Aminopropyl)-2-methylindolizine, 3-(3-Aminopropyl)-2-methyl-5,6,7,8-tetrahydroindolizine, and their N -alkyl derivatives.

The syntheses and a preliminary pharmacological evaluation of some aminopropylindolizines and aminopropyltetrahydroindolizines are reported. All compounds showed anti-5-hydroxytryptamine, anti-histamine, and antiacetylcholine activities. Some also exhibited weak CNS activity.     

Caution in the use of boldo in herbal laxatives: a case of hepatotoxicity

A case is reported in which a several-fold increase in transaminases and gamma-GT was detected in an elderly male patient with fatty liver. The patient was regularly taking a mixture of herbal products, used as a laxative, for a number of years, with no alteration of blood chemistry until 6 months before the present observation. However, the composition of the mixture had been modified by the manufacturer in the past 5 months, with addition of boldo leaf extracts. Transaminases promptly returned to normal after withdrawal of the laxative. It is concluded that boldo leaf extracts might be hepatotoxic, at least in elderly patients with fatty liver.