Temporal trend of drug-resistance and APOBEC editing in PBMC genotypic resistance tests from HIV-1 infected virologically suppressed individuals.

BACKGROUND: We aimed at evaluating the temporal trend of drug-resistance and APOBEC editing from HIV-DNA genotypic resistance tests (GRT) in virologically suppressed individuals. MATERIAL AND METHODS: Major resistance mutations (MRM), genotypic susceptibility score (GSS) for the current regimen and APOBEC-related mutations (APO-M) were evaluated. Potential changes in trends of MRM and APO-M over-time were assessed and predictors of MRM detection or sub-optimal GSS (GSS<2) at HIV-DNA-GRT were estimated through logistic regression analyses. RESULTS: Among the 1126 individuals included, 396 (35.2%) harboured at least one MRM (23.4% to NRTI, 18.8% to NNRTI, 7.7% to PI and 1.4% to INSTI [N=724]); 132 (12.3%) individuals showed a GSS <2. APO-M and stop codons were found in 229 (20.3%) and 105 (9.3%) individuals, respectively. APO-DRMs were found in 16.8% of individuals and were more likely observed in those individuals with stop codons (40.0%) compared to those without (14.4%, P<0.001). From 2010 to 2021 no significant changes of resistance or APO-M were found. Positive predictors of MRM detection at HIV-DNA GRT were drug abuse, subtype B infection, and a prolonged and complex treatment history. Perinatal infection and having at least 2 stop codons were associated with a current suboptimal regimen. CONCLUSIONS: In virologically suppressed individuals, resistance in HIV-DNA and the extent of APOBEC editing were generally stable in the last decade. A careful evaluation of APOBEC editing might be helpful to improve the reliability of HIV-DNA GRT. Further investigations are required to understand how to apply the estimation of APOBEC editing in refining genotypic evaluation.

HIV MDR is still a relevant issue despite its dramatic drop over the years.

OBJECTIVES: To evaluate the prevalence and therapeutic relevance of drug resistance among isolates from ART-experienced HIV-1-infected patients over the past two decades in Italy. METHODS: Dynamics of resistance to one, two and three or more antiretroviral classes were evaluated from 1999-2018. Virological success (VS) after the latest therapy switch was evaluated according to cumulative class resistance and cumulative genotypic susceptibility score (Stanford HIV_DB algorithm). RESULTS: Among 13 663 isolates (from 6739 patients), resistance to at least one drug class decreased sharply from 1999 to 2010 (≤2001, 84.6%; 2010, 43.6%; P < 0.001), then remained relatively constant at ∼40% during 2010-18, with the proportion of resistance to three or more classes also stable (∼5%). After 2008, integrase inhibitor resistance slightly increased from 5.6% to 9.7% in 2018 and contributed to resistance, particularly in isolates with resistance to three or more classes (one class, 8.4%; two classes, 15.3%; three or more classes, 34.7%, P < 0.001). Among 1827 failing patients with an available follow-up, by 1 year after genotype-guided therapy start the probability of VS was 87.6%. Patients with cumulative resistance to three or more classes and receiving a poorly active regimen showed the lowest probability (62.6%) of VS (P < 0.001) compared with all other patients (≥81.8%). By Cox regression analysis, cumulative MDR and receiving poorly active antiretroviral regimens were associated with a lower hazard of VS compared with those without resistance. CONCLUSIONS: A dramatic drop of HIV-1 drug resistance at failure has been achieved over the last two decades in Italy; resistance to three or more classes is low but present among currently failing patients. Its management still requires a rational and careful diagnostic and therapeutic approach.

Persistent risk of HBV reactivation despite extensive lamivudine prophylaxis in haematopoietic stem cell transplant recipients who are anti-HBc-positive or HBV-negative recipients with an anti-HBc-positive donor.

The overall rate of hepatitis B virus (HBV) reactivation was evaluated in a population of 373 haematological stem cell transplant (HSCT) patients treated with lamivudine (LMV) if they were anti-HBc-positive/HBV-DNA-negative recipients or if they were HBV-negative recipients with an anti-HBc-positive donor. The incidence of HBV reactivation was calculated in two groups of autologous (auto) or allogeneic (allo) HSCT patients who were stratified according to their HBV serostatus. The former group included 57 cases: 10 auto-HSCT and 27 allo-HSCT anti-HBc-positive recipients, two auto-HSCT and three allo-HSCT inactive carriers, and 15 allo-HSCT recipients with an anti-HBc-positive donor. Forty-seven (82.4%) patients in this group received LMV prophylaxis (the median (interquartile range, IQR) of LMV treatment was 30 (20-38) months). The second group consisted of 320 anti-HBc-negative auto-HSCT and allo-HSCT recipients with anti-HBc-negative donors. None of these patients received any prophylaxis. Two patients in the first group and two in the second group experienced reactivation of HBV infection, with an incidence of 3.5% (95% CI 0.4-12.1%) and 0.6% (95% CI 0.1-2.2%), respectively. Only one out of four reactivated patients was LMV-treated. The cumulative probability of HBV reactivation at 6 years from HSCT was 15.8% (95% CI 15.2-16.4%). Three of four viral isolates obtained from the HBV-reactivated patients harboured mutations in the immune-active HBsAg-region. In a HSCT population carefully evaluated for HBV prophylaxis, a risk of HBV reactivation persisted in the group of patients who were not LMV-treated. Only one LMV-treated patient experienced reactivation of HBV with a resistant HBV isolate.

Histological and ultra-structural effects of rapid muscle expansion through intramuscular administration of carbon dioxide: an intra-operative study in an animal model.

INTRODUCTION: Carbon dioxide (CO2) therapy refers to trans-cutaneous or sub-cutaneous administration of CO2 for therapeutic purposes, and recent studies have pointed out that it produces a vasodilation effect after it is locally injected, which helps amplify the reconstructive potentiality of an expanded-muscle flap. MATERIALS AND METHODS: Thirty male Wistar rats, weighting between 350 and 400 g, were randomly divided into three groups of 10. In the first group, single intra-operative rapid expansion was carried out under the right latissimus dorsi muscle. In the second group, for five days prior to surgery, a pre-treatment with intramuscular injections of CO2 was performed. The third group served as controls. For each group, the latissimus dorsi muscle was fixed as soon possible after mice died, and ultrathin sections of it examined with transmission electron microscope. RESULTS: In the treated group, the majority of expanded muscles showed a normal striation pattern, whereas a few fibers showed mild disorganization of the myo-filaments in the sarcomeres, which appeared overstretched (average 2.37 µm). CONCLUSIONS: This evidence could demonstrate a greater capacity of muscle recovery after treatment by CO2 expansion.

Influence of rotational speed on the cyclic fatigue of Mtwo instruments.

AIM: To evaluate the effect of rotational speed on cyclic fatigue of Mtwo nickel-titanium files. METHODOLOGY: A total of 120 new Mtwo rotary instruments sizes 10, 0.04 taper; 15, 0.05 taper; 20, 0.05 taper; and 25, 0.06 taper were randomly divided into three groups on the basis of the rotational speed used to shape nine standardized simulated canals: group A = 350 rpm; group B = 250 rpm; group C = 150 rpm. Each group consisted of 40 instruments, 10 for every size. The average preparation time (in seconds) and the average correlated numbers of cycles to instrument (NCI) the nine standardized canals were recorded for each file. The resistance to cyclic fatigue was determined by counting numbers of cycles to failure (NCF) with a rotational speed of 300 rpm in a 60° curve with a 5-mm radius. Data were analysed by two-way anova. RESULTS: Preparation time was significantly longer at 150 rpm than at 250 or 350 rpm. The average number of cycles needed for each file to instrument nine standardized canals was significantly higher at 350 and 250 rpm, than at 150 rpm. There were no significant differences in the NCF (P > 0.05) between A, B and C groups for instruments of the same size. CONCLUSIONS: Speed did not affect the cyclic fatigue of Mtwo instruments with the same size and taper. Preparation time was shorter at 350 or 250 rpm rather than at 150 rpm. However, there was no significant difference between 350 and 250 rpm rotational speed, neither in the preparation time of simulated canals nor in the resistance to fatigue fracture.

El sistema rotatorio Mtwo NiTi para preparación de conductos / Mtwo Niti rotary system for root canal preparation

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A case of small-cell gastric carcinoma with an adenocarcinoma component and hepatic metastases: treatment with systemic and intra-hepatic chemotherapy.

Primary small-cell carcinoma (SmCC) of the stomach is a rare neoplasm with a poor prognosis and unclear histogenesis: to date, only 50 cases, including ours, have been reported in the literature. In the World Health Organization gastrointestinal tumours' classification, SmCC of the stomach has been recognized as an 'independent entity affecting the stomach'. In this paper, the authors present a clinical case and the surgical treatment of an adult with a SmCC of the stomach associated with gastric adenocarcinoma. After laparotomy, a large neoplasm with locoregional extension and multiple liver metastases were found. A palliative resection, subtotal gastrectomy, was performed, followed by systemic and intra-hepatic chemotherapy: computed tomography scan demonstrated a marked response, but the patient died 15 months after the operation. A review of the literature showed that the diagnosis of gastric SmCC is based on immunohistochemical findings. Our experience confirmed the high aggressiveness of this neoplasm, which is generally diagnosed in advanced stage and is unresponsive to chemotherapy, but the combined use of systemic and intra-hepatic chemotherapy shows an acceptable result in a palliative care perspective.

Pulp-dissolving ability of several endodontic irrigants: a spectrophotometric evaluation.

The aim of this study is to test the dissolving action of different concentrations of NaOCl (0.5%, 2.5% and 5%) and EDTA 17% on bovine pulp tissue, testing solutions at three different times (1, 5, 10 minutes), 100 mg.of dental pulp placed into a becker with 20 ml of solution was evaluated using a spectrophotometer. A solution of NaOCl 5% showed the highest solubility value. Samples treated with 2.5% NaOCl showed solubility values of 0.98 in one minute, 29.1 in five minutes and 57 in ten minutes. Groups treated with NaOCl 0.5% showed small dissolving modifications. 17% EDTA showed the lowest values of solubility. 5% NaOCl solution showed highest pulp dissolving ability, even a 2.5% solution showed high dissolving ability but in longer times.

Evaluation of tumour-infiltrating CD4+CD25+FOXP3+ regulatory T cells in human cutaneous benign and atypical naevi, melanomas and melanoma metastases.

BACKGROUND: CD4+CD25+FOXP3+ regulatory T cells (Tregs) are thought to induce immunotolerance in melanoma. They have not yet been investigated in the entire spectrum of melanocytic cutaneous lesions within a tumour site. OBJECTIVES: To evaluate CD4+CD25+FOXP3+ Tregs among tumour-infiltrating lymphocytes in cutaneous melanocytic lesions. METHODS: We analysed 128 lesions (10 benign junctional common naevi, 10 benign compound common naevi, 10 compound Spitz naevi, 10 junctional atypical naevi, 20 compound atypical naevi, 20 radial growth phase melanomas, 30 vertical growth phase melanomas and 18 melanoma metastases). Tregs were identified by CD25-FOXP3 double immunostains. RESULTS: This study indicates that CD4+/CD25+FOXP3+ Tregs are present in all groups of lesions. Junctional atypical naevi, compound atypical naevi and radial growth phase melanomas showed the highest percentages of CD4+CD25+FOXP3+ Tregs (junctional atypical naevi vs. junctional common naevi, compound common naevi, compound Spitz naevi, melanoma metastases: P < 0.0001; junctional atypical naevi vs. vertical growth phase melanomas: P = 0.001; compound atypical naevi vs. junctional common naevi, compound common naevi: P < 0.0001; compound atypical naevi vs. compound Spitz naevi, melanoma metastases: P = 0.002; compound atypical naevi vs. vertical growth phase melanomas: P = 0.02; radial growth phase melanomas vs. junctional common naevi, compound common naevi, compound Spitz naevi, melanoma metastases: P < 0.0001; radial growth phase melanomas vs. vertical growth phase melanomas: P = 0.008). CONCLUSIONS: The strong prevalence of CD25+FOXP3+ Tregs both in junctional and compound atypical naevi and radial growth phase melanomas, suggests that they induce immunotolerance early during melanoma genesis, favouring melanoma growth. Their evaluation within a tumour site could be useful for prognostic and therapeutic purposes.

Influence of a brushing working motion on the fatigue life of NiTi rotary instruments.

AIM: To evaluate the cyclic fatigue resistance of Mtwo NiTi rotary instruments when used with a brushing or no-brushing action in oval root canals. METHODOLOGY: Cyclic fatigue testing of instruments was performed in tapered artificial canals with a 5 mm radius of curvature and an angle of curvature of 60 degrees. Twenty Mtwo instruments for each size were selected and divided into two groups: group A = 10 instruments used with a no-brushing motion (control group); group B = 10 instruments used with a brushing-milling action. Each Mtwo instrument was used for cleaning and shaping 10 oval root canals. In all 80 instruments were rotated until fracture occurred and the number of cycles to failure (NCF) recorded. Data were analysed by one-way ANOVA, Tukey HSD test and independent sample t-test to determine any statistical difference; the significance was determined at the 95% confidence level. RESULTS: No statistically significant reduction of NCF between instruments used with a no-brushing motion (group A) and instruments used with a brushing motion (group B) were apparent except for size 25, 0.06 taper. Mtwo size 10, 0.04 taper, size 15, 0.05 taper, size 20, 0.06 taper, size 25, 0.06 taper instruments had a decrease in life span of 1%, 0.5%, 8% and 19%, respectively. CONCLUSIONS: Fatigue life of instruments of larger size could be reduced by using them with a lateral brushing or pressing movement. However, each file was successfully operated without intracanal failure, demonstrating that Mtwo rotary instruments can be used safely in a brushing action in simulated clinical conditions up to 10 times in oval canals.

[Primary cardiac echinococcosis]. / Echinococcosi cardiaca primitiva.

Cardiac echinococcosis is very rare, even in endemic regions. It has been reported that cardiac involvement is seen in about 0.5-3% of human echinococcosis cases. A case of cardiac isolated hydatid desease that caused sporadic syncopal episodes is illustrated.

Cyclic fatigue resistance and three-dimensional analysis of instruments from two nickel-titanium rotary systems.

AIM: To determine how instrument design affects the fatigue life of two nickel-titanium (Ni-Ti) rotary systems (Mtwo and ProTaper) under cyclic fatigue stress in simulated root canals. METHODOLOGY: Cyclic fatigue testing of instruments was performed in stainless steel artificial canals with radii of curvature of 2 or 5 mm and an angle of curvature of 60 degrees . A total of 260 instruments were rotated until fracture occurred and the number of cycles to failure were recorded. The morphology of Ni-Ti rotary instruments was investigated by measuring the volume of millimetre slices of each instrument size starting from the tip to the shank by means of microCT analysis. The fracture surface of three representative samples of each size was analysed by scanning electron microscopy (SEM). Data were analysed by one-way anova, Holm t-test, paired t-test and linear regression; the significance was determined at the 95% confidence level. RESULTS: Cycles to failure significantly decreased as the instrument volume increased for both the radii of curvature tested (P < 0.01). The radius of curvature had a statistically significant influence on the fatigue life of the instruments (P < 0.05). Larger instruments underwent fracture in less time under cyclic stress than smaller ones. SEM evaluation showed typical features of fracture through fatigue failure. CONCLUSIONS: The metal volume in the point of maximum stress during a cyclic fatigue test could affect the fatigue life of Ni-Ti rotary instruments. The larger the metal volume, the lower the fatigue resistance.

A comparison of cyclic fatigue between used and new Mtwo Ni-Ti rotary instruments.

AIM: To evaluate the cyclic fatigue of Mtwo Ni-Ti rotary instruments after controlled clinical use in molar teeth. METHODOLOGY: Twenty Mtwo instruments of each size were selected and divided into two groups: group A consisted of 10 new instruments (control group); group B consisted of 10 used instruments. Each instrument in group B was used to clean and shape 10 root canals of molar teeth in patients. Cyclic fatigue testing of instruments was performed in tapered artificial canals with a 5-mm radius of curvature and a 60 degrees angle of curvature. In all 140, instruments were rotated until fracture and the number of cycles to failure was recorded. Data were analysed using one-way anova, Tukey's HSD test and independent sample t-test to determine any statistical difference; the significance was determined at the 95% confidence level. RESULTS: A reduction of cycles to failure between new (group A) and used (group B) instruments was apparent. A statistically significant difference (P < 0.05) was noted between instruments of groups A and B in all sizes with the exception of size 40, 0.04 taper. CONCLUSIONS: Clinical use significantly reduced cyclic fatigue resistance of Mtwo rotary instruments when compared with an unused control group. However, all the instruments had minimal instrument fatigue when discarded after controlled clinical use.

SEM evaluation of canal wall dentine following use of Mtwo and ProTaper NiTi rotary instruments.

AIM: To compare using scanning electron microscopy (SEM) root canal walls following instrumentation in vitro with two different rotary NiTi instruments. The hypothesis was that no difference should be observable between the experimental groups in terms of debris on canal walls and surface morphology. METHODOLOGY: Twenty-four single-rooted human teeth were selected. Two types of NiTi instruments were used, Mtwo (Sweden & Martina, Padova, Italy) and ProTaper (Dentsply Maillefer, Ballaigues, Switzerland). Irrigation for both groups was performed after each instrument change with 5% NaOCl, 3% H2O2 and 17% EDTA solutions. Three different areas (coronal, middle and apical thirds) of the root canal were evaluated using SEM. The canal wall of each sample was assessed and compared using a predefined scale of four parameters, namely, smear layer, pulpal debris, inorganic dentine debris, surface profile. Data were analysed statistically using the Kruskal-Wallis test (anova). RESULTS: A statistically significant difference (P < 0.01) was found between the apical third and the middle and coronal thirds for both groups. No difference was observable between instrumentation groups. In the apical third canal walls were often contaminated by inorganic debris and by smear layer. In the apical third, the surface profile was affected by uninstrumented regions, comprising dentine depressions and grooves in which predentine was still visible. CONCLUSION: Both instruments produced a clean and debris-free dentine surfaces in the coronal and middle thirds, but were unable to produce a dentine surfaces free from smear layer and debris in the apical third. The presence of deep grooves and depression on dentine walls in the apical third may well explain the presence of less-instrumented areas.

[Today's trends in endodontics]. / Gli attuali orientamenti in endodonzia.

Some new means and innovative operative techniques in the field of endodontics are presented that have been developed and perfected over time. Instruments such as the operative microscope, ultrasonography, miniaturised tools for surgery, new materials such as mineral trioxide aggregate (MTA) should become part of the daily routine of the endodontist. For example, the operative microscope plays a role in diagnosing pulp damage after removal of deep caries; in orthograde endodontics in diagnosing perforations; alongside ultrasonography in the search for calcified canals, removal of pins (screw-type, fibre, etc.) and also in positioning MTA. In all these cases, the operative microscope is used discontinuously, alternating working and observation phases. In endodontic surgery, the operative microscope is on the contrary used continuously, enabling the surgeon to observe details of anatomy of the root apex at high magnification, and above all enabling positioning of canal obturation with perfect seal. It is clear that the operative microscope used in all surgical phases will enable complete management of the operation and facilitate the endodontist in observing the operative field, otherwise very difficult to control.

Some unusual clinical cases on root anatomy of permanent maxillary molars.

Three case reports are presented concerning permanent maxillary molars with fusion of two buccal roots. The morphology is peculiar because it is characterized by the mesiobuccal canal merging into the distobuccal canal in their apical one-third. A thorough review of the literature failed to reveal any previously reported description of permanent maxillary molars with this arrangement of canals, with the exception of a recently reported epidemiological study.

[Immunological phenomena in pulpal-periapical lesions]. / Fenomeni immunitari nelle lesioni pulpo-periapicali.

Based on an extensive review of the literature, the authors examine immunological reactions in pulpal and periapical lesions. Although it has been known for some time that bacterial infection causes this pathology, attention has been recently focused on immunological factors in the ambit of the phlogistic process. The present study examines the correlation between the latter and the type of extent of antigenic response, focusing attention on their important role in the phenomena of osteoclastic activation and inhibition of bone repair. From a physiological point of view there are few inflammatory cells in dental pulp, like macrophages and T lymphocytes. When the pulp comes into contact with the antigenic substance it activates a specific and aspecific immune response: the form through the activation of B and T lymphocytes, and the latter through the action of LPS, PMN and complement. An important role in the immune response is played by the cytokines which are able to regulate the intensity and duration of the immune response against potential pathogenic agents. It was initially thought that these were only produced by lymphocytes and as a result they were known as lymphokines. Later it was observed that other cell populations were also able to produce them. Phlogosis of the periapex starts before the pulp is fully necrotic. Tissue detritus and products of bacterial derivation escape through the numerous endoparadontal communication pathways and stimulate an inflammatory response by the vascular system of the parodontal ligament. The concomitant immune reaction occurs due to the tendency to block and restrict the inflammation to the radicular channels, thus preventing its diffusion.(ABSTRACT TRUNCATED AT 250 WORDS)

[Morphological characteristics of spreaders]. / Indagine sulle caratteristiche morfologiche degli spreaders.

Different kinds of spreaders can be used in the lateral condensation technique. Selection of condensation instruments must not be empirical because the quality of the filling partly depends on the compatibility between the spreader and the prepared root canal. The shape of the instrument should allow for easy insertion to the proper length without any interference. On these bases the Authors evaluated and compared the morphology of some spreaders, and gave data and directions how to rationally use these instruments in clinical practice.