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The increasing necessity for solo surgery in plastic and reconstructive microsurgery is driven by contemporary challenges such as a growing and aging population, a shortage of qualified assistants, and the ongoing recovery efforts from COVID-19. Historically limited to remote or exceptional circumstances, solo surgery is now more frequently performed due to these evolving factors. Technological advancements, including robotics, play a crucial role in facilitating this transition and supporting the trend toward self-sufficiency in surgical practice. This article presents practical strategies for executing solo surgery based on our team's experience and a review of current literature. Key considerations discussed include patient positioning and optimizing surgical workflows. By presenting these recommendations and techniques, derived from both practical experience and literature, we highlight that plastic surgeons and microsurgeons can maintain consistently high standards of surgical care, even when operating independently., Competing Interests: The authors have no financial interest to declare in relation to the content of this article., (Copyright © 2025 The Authors. Published by Wolters Kluwer Health, Inc. on behalf of The American Society of Plastic Surgeons.)

Colorectal cancer represents 10% of all the annual tumors diagnosed worldwide, being often not timely diagnosed, because its symptoms are typically lacking or very mild. Therefore, it is crucial to develop and validate innovative low-invasive techniques to detect it before becoming intractable. To this aim, a device equipped with nanostructured gas sensors has been employed to detect the airborne molecules of blood samples collected from healthy subjects, and from colorectal cancer affected patients at different stages of their pre- and post-surgery therapeutic path. Data was scrutinized by using statistical standard techniques to highlight their statistical differences, and through principal component analysis and support vector machine to classify them. The device was able to readily distinguish between the pre-surgery blood samples (i.e., taken when the patient had cancer), and the ones up to three years post-surgery (i.e., following the tumor removal) or the ones from healthy subjects. Finally, the correlation of the sensor responses with the patient/healthy subject's gender was investigated, resulting negligible. These results pave the path toward a clinical validation of this device to monitor the patient's health status by detecting possible relapses, to parallel to clinical follow-up protocols.

Background: The success of total ankle arthroplasty (TAA) involves objective and subjective parameters such as joint motion and implant survival. Patient-reported outcome measures provide key context from the patient's perspective so that we can evaluate the results of these procedures. The Forgotten Joint Score (FJS) has demonstrated validity and reliability in other arthroplasties, but the existing evidence on the FJS in TAA is very limited., Question/purposes: We aimed to assess the reliability, validity, responsiveness to change, and floor and ceiling effects, as well as to perform a gender analysis, of the FJS in patients who had undergone TAA., Methods: A prospective, observational study enrolled patients who underwent TAA at our institution between June 2021 and May 2023. The three-component TAA was performed through an anterior approach. A total of 120 patients (mean age 62 ± 10 years) participated, which was 83% (120 of 145) of those eligible, and 77% (92 of 120) of those patients underwent follow-up assessments at 12 ± 1 months after surgery. A cross-culturally adapted and validated version of the FJS in our language (Italian) was used. Additionally, three other commonly used patient-reported outcome measures for TAA were administered: the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, the Manchester-Oxford Foot Questionnaire (MOXFQ), and the VAS., Results: Reliability was demonstrated by excellent internal consistency (Cronbach α coefficient = 0.95) and excellent test-retest reliability (intraclass correlation coefficient = 0.99; standard error of measurement = 2.7). Robust validity was observed, in that the FJS had a strong correlation with the AOFAS ankle-hindfoot score, the MOXFQ, and the VAS (Pearson and Spearman values consistently above 0.7 or below -0.7). Responsiveness to change was observed between 6 and 12 months (Cohen d = 0.37). Low ceiling and floor effects at both 6 months (4% and 3% reached top and bottom scores, respectively) and 12 months (10% and 0% reached top and bottom scores, respectively) were demonstrated, below the recommended 15% interpretability threshold. No association between outcomes and patient gender was observed., Conclusion: Test properties and validity of the FJS were strong in patients who underwent TAA. It seems suitable for clinical use, although future studies should seek to replicate or refute our findings in other patient populations., Level of Evidence: Level II, therapeutic study., Competing Interests: Each author certifies that there are no funding or commercial associations (consultancies, stock ownership, equity interest, patent/licensing arrangements, etc.) that might pose a conflict of interest in connection with the submitted article related to the author or any immediate family members. All ICMJE Conflict of Interest Forms for authors and Clinical Orthopaedics and Related Research® editors and board members are on file with the publication and can be viewed on request., (Copyright © 2024 by the Association of Bone and Joint Surgeons.)

Subtalar distraction arthrodesis (SDA) is a surgical procedure designed to treat hindfoot deformities associated with isolated subtalar joint arthritis. In 1996, Fitzgibbons was the first to observe that, in some cases, hindfoot fusion appeared to be associated with the development of tibiotalar valgus tilt. Since then, few studies have addressed this issue. Given that hindfoot fusion can be performed using various techniques, this study investigated the potential tibiotalar joint frontal or sagittal modifications resulting from the modified Grice-Green technique. All the consecutive patients who underwent the modified Grice-Green procedure were included. The patient records were reviewed to extract demographic data. Weight-bearing foot and ankle radiographs were assessed to measure the talar tilt angle and the tibiotalar ratio on the same picture archiving and communication system by three independent observers. A total of 69 patients met the criteria for inclusion. The mean talar tilt showed no substantial changes, since the increase from 1.46 ± 1.62 preoperatively to 1.93 ± 2.19 at a minimum of 8 months postoperatively was not statistically significant ( p = 0.47). The average preoperative tibiotalar ratio significantly increased from 33.4 ± 4.4% to 35 ± 4% postoperatively ( p = 0.007), although remaining within the normal range, indicating a possible realignment of the posterior facet of the subtalar joint. In conclusion, this study highlights the effectiveness of the modified Grice-Green procedure in achieving a favorable realignment without impacting the ankle joint, particularly regarding tibiotalar valgus tilt.

Chemoresistive nanostructured gas sensors are employed in many diverse applications in the medical, industrial, environmental, etc. fields; therefore, it is crucial to have a device that is able to quickly calibrate and characterize them. To this aim, a portable, user-friendly device designed to easily calibrate a sensor in laboratory and/or on field is introduced here. The device comprises a small hermetically sealed chamber (containing the sensor socket and a temperature/humidity sensor), a pneumatic system, and a custom electronics controlled by a Raspberry Pi 4 developing board, running a custom software (Version 1.0) whose user interface is accessed via a multitouch-screen. This device automatically characterizes the sensor heater in order to precisely set the desired working temperature, it acquires and plots the sensor current-to-voltage and Arrhenius relationships on the touch screen, and it can record the sensor responses to different gases and environments. These tests were performed in dry air on two representative sensors based on widely used SnO 2 material. The device demonstrated the independence of the Arrhenius plot from the film applied voltage and the linearity of the I-Vs, which resulted from the voltage step length (1-30 min) and temperature (200-550 °C).

Colorectal cancer (CRC) represents 10% of the annual tumor diagnosis and deaths occurring worldwide. Given the lack of specific symptoms, which could determine a late diagnosis, the research for specific CRC biomarkers and for innovative low-invasive methods to detect them is crucial. Therefore, on the basis of previously published results, some volatile organic compounds (VOCs), detectable through gas sensors, resulted in particularly promising CRC biomarkers, making these sensors suitable candidates to be employed in CRC screening devices. A new device was employed here to analyze the exhalations of blood samples collected from CRC-affected patients at different stages of their pre- and post-surgery therapeutic path, in order to assess the sensor's capability for discriminating among these samples. The stages considered were: the same day of the surgical treatment (T1); before the hospital discharge (T2); after one month and after 10-12 months from surgery (T3 and T4, respectively). This device, equipped with four different sensors based on different metal-oxide mixtures, enabled a distinction between T1 and T4 with a sensitivity and specificity of 93% and 82%, respectively, making it suitable for clinical follow-up protocols, patient health status monitoring and to detect possible post-treatment relapses.

Nowadays, most medical-diagnostic, environmental monitoring, etc. devices employ sensors whose fabrication reproducibility and response repeatability assessment are crucial. The former consists of large-scale sensor manufacture through a standardized process with almost identical morphology and behavior, while the latter consists of giving the same response upon repeating the same stimulus. The thermo-activated chemoresistive sensors, which change their conductance by interacting with the molecules composing the surrounding gas, are currently employed in many devices: in particular, thick-film (SnTiNb)O 2 nanosensors were demonstrated to be particularly suitable in the medical and biological fields. Therefore, a set of thirteen of them, randomly selected from the same screen-printing deposition, were laboratory tested, and the outcomes were statistically analyzed in order to assess their consistency. At first, the working temperature that maximized both the sensor sensitivity and response repeatability was identified. Then, the sensors were subjected to different gas concentrations and humidities at this optimal working temperature. It resulted in the (SnTiNb)O 2 nanosensors detecting and discriminating CO concentrations as low as 1 ppm and at high humidity degrees (up to 40%) with high repeatability since the response relative standard error ranged from 0.8 to 3.3% for CO and from 3.6 to 5.4% for water vapor.

This study aimed to identify and characterise indoor sources of particulate matter (PM) in domestic environments. 74 inhabited apartments located in the urban area of Gela (Sicily, Italy), close to a refinery, and in three villages of the hinterland were evaluated, in real-world conditions, for the elemental composition of PM 2.5 . The samples were collected simultaneously inside and outside each apartment for 48 h. In addition, two of the apartments were simultaneously studied for four weeks. The elemental composition of PM 2.5 was determined by applying a chemical fractionation procedure followed by inductively-coupled plasma spectrometry analysis, with both optical emission and mass detection. The extractable, more bio-accessible fraction ( ext ), and the mineralised residual fraction ( res ) of each element were determined, thus increasing the selectivity of elements as source tracers. Indoor air in the considered apartments was affected by both outdoor pollution and specific indoor emission sources. The behaviour of each source was studied in detail, identifying a reliable tracer: Ti res for soil, As ext for industrial sources, V ext for heavy oil combustion, Ce for cigarette smoking and Mo for the use of vacuum dust cleaners. As ext and V ext showed an excellent infiltration capacity, while the concentration of Ti res was affected by a low infiltration capacity and by the contribution of particles re-suspension caused by the residents' movements. In the case of Ce and Mo, indoor concentrations were much higher than outdoor with a high variability among the apartments, due to the inhabitants' habits concerning cigarette smoke and use of electric appliances. To test the overall effect of the concomitant exposure to the identified sources on Wh12 M and on DDA, a WQS analysis was conducted. Cigarette smoking and heavily oil combustion driven the Wh12 M odds increase, while the DDA odds increase was mainly driven by heavily oil combustion and the use of vacuum dust cleaners., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Inc.)

The detection of volatile organic compounds (VOCs) exhaled by human body fluids is a recent and promising method to reveal tumor formations. In this feasibility study, a patented device, based on nanostructured chemoresistive gas sensors, was employed to explore the gaseous exhalations of tumoral, immortalized, and healthy cell lines, with the aim of distinguishing their VOC patterns. The analysis of the device output to the cell VOCs, emanated at different incubation times and initial plating concentrations, was performed to evaluate the device suitability to identify the cell types and to monitor their growth. The sensors ST25 (based on tin and titanium oxides), STN (based on tin, titanium, and niobium oxides), and TiTaV (based on titanium, tantalum and vanadium oxides) used here, gave progressively increasing responses upon the cell density increase and incubation time; the sensor W11 (based on tungsten oxide) gave instead unreliable responses to all cell lines. All sensors (except for W11) gave large and consistent responses to RKO and HEK293 cells, while they were less responsive to CHO, A549, and CACO-2 ones. The encouraging results presented here, although preliminary, foresee the development of sensor arrays capable of identifying tumor presence and its type.

Background and Aim: The roles of physicians and nurses have undergone profound changes in recent years, becoming more complex and creating, at times, overlapping competencies with consequent negative repercussions in terms of problems related to autonomy and inter-professional collaboration. The study aims to detect the opinions of intensive care physicians and nurses with respect to their skills and role expectations., Method: The study was conducted in two phases. In the first phase, with a qualitative design, data were collected through the focus group. In the second phase, with quantitative design, an ad hoc questionnaire was constructed from the results of the previous phase.   Results: 3 main themes emerged from the focus groups: the need to increase the system of inter-professional collaboration; autonomy and professional role; and the need for a shared advanced training system for physicians and nurses. The results of the questionnaire show that both professionals believe that the development of advanced skills does not lead to conflicting situations and role ambiguity unless the skills are recognized and respected. Both professionals agree that it is useful and necessary to implement a shared educational pathway.   Conclusion: Inter-professional collaboration, specialist skills and specific training are a key element of the ICU teamwork.

Background: The benefit of rotator cuff repair (RCR) in patients with concurrent osteoarthritic changes remains unclear. RCR has the theoretical potential to increase the compressive force across the glenohumeral joint, further exacerbating osteoarthritis pain. The purpose of this study is to investigate pain relief and patient-reported outcomes of patients undergoing simultaneous RCR and microfracture of focal glenohumeral osteoarthritis., Methods: Thirty-four patients undergoing simultaneous RCR and microfracture were retrospectively reviewed at a minimum 1-year follow-up. Patient demographics, preoperative range of motion, functional outcomes (visual analog scale [VAS], Single Assessment Numeric Evaluation [SANE], American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form [ASES], and Simple Shoulder Test [SST]), and operative metrics were recorded. The patients were then contacted to obtain postoperative functional outcome scores (VAS, SANE, ASES, and SST)., Results: Twenty-seven patients (11 male/16 female [79%]) were evaluated at a mean follow-up of 25.8 months (range, 12-46). The average age at surgery was 64.9 years (range, 56-78). Chronic tears were more common than acute tears (57.7% vs. 42.3%). The majority of patients had a full rotator cuff tear (89%) involving a mean 1.7 ± 0.8 tendons (range, 1-3). Eighty-eight percent of the humeral lesions were Outerbridge 4 compared with 84% on the glenoid. The mean estimated involvement between the 2 groups with 38.4% ± 18.4% of the humeral head involved and 34.6% ± 18.4% of the glenoid involved. PRO scores improved postoperatively with a reduction in mean VAS (6.6-2.0, P < .01), SANE (33.8-79.8, P < .01), ASES (38.0-80.9, P < .01), and SST (3.07-9.70, P < .01) scores. Cumulatively, only 52% (14/27) of the patients improved, however, by the MCID for all collected PROs., Conclusions: Our results demonstrate modest improvements in postoperative pain and functional scores at a minimum of 1-year follow-up in a cohort of patients who have undergone RCR and glenohumeral microfracture. In cases of small focal lesions of full-thickness cartilage loss, RCR with microfracture is a reasonable treatment option; however, patients should be counseled on expectations accordingly., (Copyright © 2021 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

User-friendly, low-cost equipment for preventive screening of severe or deadly pathologies are one of the most sought devices by the National Health Services, as they allow early disease detection and treatment, often avoiding its degeneration. In recent years more and more research groups are developing devices aimed at these goals employing gas sensors. Here, nanostructured chemoresistive metal oxide (MOX) sensors were employed in a patented prototype aimed to detect volatile organic compounds (VOCs), exhaled by blood samples collected from patients affected by colorectal cancer and from healthy subjects as a control. Four sensors, carefully selected after many years of laboratory tests on biological samples (cultured cells, human stools, human biopsies, etc.), were based here on various percentages of tin, tungsten, titanium, niobium, tantalum and vanadium oxides. Sensor voltage responses were statistically analyzed also with the receiver operating characteristic (ROC) curves, that allowed the identification of the cut-off discriminating between healthy and tumor affected subjects for each sensor, leading to an estimate of sensitivity and specificity parameters. ROC analysis demonstrated that sensors employing tin and titanium oxides decorated with gold nanoparticles gave sensitivities up to 80% yet with a specificity of 70%.

Despite the great progress in screening techniques and medical treatments, colorectal cancer remains one of the most widespread cancers in both sexes, with a high death rate. In this work, the volatile compounds released from human colon cancer tissues were detected by a set of four different chemoresistive sensors, made with a nanostructured powder of metal-oxide materials, inserted into an innovative patented device. The sensor responses to the exhalation of a primary cancer sample and of a healthy sample (both of the same weight, collected during colorectal surgery from the intestine of the same patient) were statistically analyzed. The sensors gave reversible, reproducible, and fast responses for at least one year of continuous use, making them quite superior in respect to the existing diagnostic methods. Preliminary results obtained using principal component analysis of the sensor responses to samples removed from 13 patients indicate that the nanostructured sensors employed in this study were able to distinguish between healthy and tumor tissue samples with coherent responses (the discrimination power of the most sensitive sensor was about 17%), highlighting a strong potential for clinical practice., Competing Interests: The authors declare no conflicts of interest.

Preventive screening does not only allow to preemptively intervene on pathologies before they can harm the host; but also to reduce the costs of the intervention itself; boosting the efficiency of the NHS (National Health System) by saving resources for other purposes. To improve technology advancements in this field; user-friendly yet low-cost devices are required; and various applications for gas sensors have been tested and proved reliable in past studies. In this work; cell cultures and blood samples have been studied; using nanostructured chemoresistive sensors; to both verify if this technology can reliably detect tumor markers; and if correlations between responses from tumor line metabolites and the screening outcomes on human specimens could be observed. The results showed how sensors responded differently to the emanations from healthy and mutant (for cells) or tumor affected (for blood) samples, and how those results were consistent between them, since the tumoral specimens had higher responses compared to the ones of their healthy counterparts. Even though the patterns in the responses require a bigger population to be defined properly; it appeared that the different macro-groups between the same kind of samples are distinguishable from some of the sensors chosen in the study; giving promising outcomes for further research.

We studied the size distribution of ions (Cl - , NO 3 - , SO 4 = , Na + , NH 4 + , K + , Mg ++ , Ca ++ ) and elements (As, Ba, Cd, Co, Cs, Cu, Fe, Li, Mn, Ni, Pb, Rb, Sb, Se, Sn, Sr, Ti, Tl, V, Zn) during the winter and summer seasons of seven consecutive years (2008-2014) in an area of the Po Valley (Northern Italy) characterised by industrial, agricultural and urban settings. The study included the collection and analysis of 41 series of size-segregated samples (MOUDI sampler, 10 stages, cut sizes from 0.18 to 18 μm). Ions were analysed by ion chromatography; elemental analysis was carried out by ICP-MS, by applying a chemical fractionation method able to increase the selectivity of PM source tracers. Our results indicate that important winter/summer variations occurred in both the concentration and size distribution of most PM components. These variations were explained in terms of variations in the strength of the prevailing sources of each component. The contribution of biomass burning for domestic heating was highlighted by the well-known tracer K + but also by the soluble fraction of Rb, Cs and Li. Biomass burning contribution to atmospheric PM was mostly contained in the fine fraction, with a broad size-distribution from 0.18 to 1.8 μm. This source also appreciably increased the concentration of other elements in fine PM (As, Cd, Co, Mn, Pb, Sb, Sn). A few PM components (tracers of sea-spray, brake lining and some industries) did not show marked seasonal variations in concentration and size distribution. However, during winter, for brake lining and industry tracers we observed an upward shift in the dimension of fine particles and a downward shift in the dimension of coarse particles, due to the ageing of the air masses., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Numerous reports have described players in professional American sports leagues who have been sidelined with a deep vein thrombosis (DVT) or a pulmonary embolism (PE), but little is known about the clinical implications of these events in professional athletes., Purpose: To conduct a retrospective review of injury reports from the National Hockey League (NHL), Major League Baseball (MLB), the National Basketball Association (NBA), and the National Football League (NFL) to take a closer look at the incidence of DVT/PE, current treatment approaches, and estimated time to return to play in professional athletes., Study Design: Descriptive epidemiology study., Methods: An online search of all team injury and media reports of DVT/PE in NHL, MLB, NBA, and NFL players available for public record was conducted by use of Google, PubMed, and SPORTDiscus. Searches were conducted using the professional team name combined with blood clot , pulmonary embolism , and deep vein thrombosis ., Results: A total of 55 venous thromboembolism (VTE) events were identified from 1999 through 2016 (NHL, n = 22; MLB, n = 16; NFL, n = 12; NBA, n = 5). Nineteen athletes were reported to have an upper extremity DVT, 15 had a lower extremity DVT, 15 had a PE, and 6 had DVT with PE. Six athletes sustained more than 1 VTE. The mean age at time of VTE was 29.3 years (range, 19-42 years). Mean (±SD) time lost from play was 6.7 ± 4.9 months (range, 3 days to career end). Seven athletes did not return to play. Players with upper extremity DVT had a faster return to play (mean ± SD, 4.3 ± 2.7 months) than those with lower extremity DVT (5.9 ± 3.8 months), PE (10.8 ± 6.8 months), or DVT with PE (8.2 ± 2.6 months) ( F = 5.69, P = .002). No significant difference was found regarding time of return to play between sports., Conclusion: VTE in professional athletes led to an average of 6.7 months lost from play. The majority of athletes were able to return to play after a period of anticoagulation or surgery. Those with an upper extremity DVT returned to play faster than those with other types of VTE. Further study is needed to look into modifiable risk factors for these events and to establish treatment and return-to-play guidelines to ensure the safety of these athletes., Competing Interests: The authors declared that they have no conflicts of interest in the authorship and publication of this contribution.

Background: Exposure to single and multiple carcinogenic metals and/or semimetals represents a major environmental risk factor for public health. In particular, children are more susceptible to environmental pollutants than adults, but specific studies are still limited. The aims of the present study were: 1) to trace the exposure and co-exposure profiles to eight known or suspected carcinogenic metals and semimetals (As, Be, Cd, Co, Cr, Ni, Pb, and Sb); and: 2) to evaluate the influence of some possible interfering/confounding factors on the exposure to these elements during childhood., Study Design: Cross-sectional study., Methods: We recruited 159 healthy Italian children attending a primary school of the urban area of Rome, Italy. Selected metals were determined by inductively coupled plasma mass spectrometry on urinary samples collected at the end of a "typical" day (one sample for each child), while information about possible confounding/interfering factors were collected via questionnaires., Results: The great part of the studied children resulted co-exposed to the monitored metals: 83.2%, 69.2%, 51.0% and 29.3% of the participants were concurrently exposed to at least two, three, four and five trace elements, respectively. Gender was the only one among the investigated variable that significantly influenced the co-exposure, with females resulting at lower risk (OR = 0.392; 95 IC = 0.156 - 0.989; p < 0.047)., Conclusions: Given the importance of protecting child's health and the risks related to the exposure to carcinogenic metals, especially when they occur simultaneously, other researches in this field are strongly recommended.

Chitosan-based polyplexes are known to traffic through lysosomes for a relatively long time, independent of the degree of deacetylation (DDA) and the number average molecular weight (Mn) of the polymer, even though both of these parameters have profound effects on polyplex stability and transfection efficiency. A better understanding of the lysosomal barrier is paramount to the rational design of vectors capable of overcoming obstacles to transgene expression. The aim of the present study was to investigate if lysosomal transit affects chitosan-based polyplex transfection efficiency in a structure-dependent (DDA, Mn) manner. Toward this end, we analyzed the effects of intracellular trafficking modifying agents on transfection efficiency and intracellular vesicular trafficking of polyplexes with different structural properties and stabilities or nucleic acid binding affinity. The use of agents that modify endosome/lysosome acidification and transit processes by distinct mechanisms and their effect on cell viability, polyplex uptake, vesicular trafficking, and transfection efficiency revealed novel and strong chitosan structure-dependent consequences of lysosomal transit. Inhibiting lysosomal transit using chloroquine significantly increased the efficiency of unstable polyplexes, while having minimal effects for polyplexes with intermediate or high stability. In parallel, specifically inhibiting the acidification of vesicles abrogated transfection for all formulations, suggesting that vesicular acidification is essential to promote transfection, most probably by facilitating lysosomal escape. These results provide novel insights into the structure-performance relationship of chitosan-based gene delivery systems.

Background: Multiple Myeloma (MM) is a neoplastic disorder characterized by clonal proliferation of malignant plasma cells (PCs). Flow cytometry is an essential tool to confirm diagnosis and evaluate minimal residual disease (MRD). This study aims at identifying new surface PC markers suitable for targeted therapy in MM and able to improve MRD detection., Methods: The expression of 82 molecules provided by the "Ninth International Workshop on Leukocyte Antigens" was analyzed by flow cytometry in 5 MM cell lines and in 20 newly diagnosed MM (NDMM) patients. Based on the antigens expression and monoclonal antibody availability, CD150, CD48, CD229, CD352, CD319, CD272, CD86, CD200 and CD184 were subsequently tested in 24 NDMM, 8 relapsed MM (RMM), 6 plasma cell leukemia (PCL) and 13 healthy subjects., Results: CD352 was less frequently expressed on NDMM than on healthy PCs; CD200 was more frequently expressed on NDMM than on RMM and healthy PCs. CD150, CD319, CD229, CD352 Mean Fluorescence Intensity (MFI) was lower in pathological than in healthy samples. The proportion of CD150-positive samples was lower in NDMM and RMM than in healthy subjects; CD86+ samples were less frequent in NDMM than in healthy subjects; CD200+ samples were more frequent in NDMM than in RMM and healthy subjects., Conclusions: CD150, CD86 and CD200 can help to identify malignant PCs; CD272, CD319, CD229, CD48 are highly expressed on all PCs and could be considered for targeted therapy. All these antigens could be added to a routine panel for PCs identification and MRD evaluation., (© 2015 International Clinical Cytometry Society.)

Clinical Scenario: Osteoarthritis (OA) is a debilitating degenerative disease affecting an estimated 27 million Americans. A systematic review found that patients with a previous history of traumatic knee injury are at increased risk of developing knee OA, regardless of specific injury. It is vital for the maintenance of quality of life for individuals affected with OA that the treatment options available be able to reduce symptoms and restore quality of living. The pain-relief benefits of traditional injection treatments are small to moderate and have a limited duration. It was found that at 2 wk postinjection that corticosteroids were more effective than hyaluronic acid (HA) injections. Autologous conditioned serum (ACS) injection is a novel treatment that has shown favorable results. However, many clinicians continue to use HA injections for reduction of symptoms in patients with osteoarthritis when the use of ACS may be more beneficial., Focused Clinical Question: For patients with knee OA, is an ACS injection more efficient at producing a reduction in symptoms than HA or a saline injection?

The optimal ratio of the polycation's amine to DNA phosphate group (N:P) for efficient polymer-based transfection always employs excess polycation versus DNA. Most of the excess polycation remains free in solution, unassociated with the polyplexes, but is essential for efficient transfection. The mechanism by which excess polycation increases transfection efficiency is not identified. We hypothesised that excess chitosan facilitates intracellular lysosomal escape of the polyplexes. We highlight here the essential role of excess chitosan by rescuing poorly transfecting low N:P ratio polyplexes, by adding free chitosan before or after polyplex addition to cells. We examined polyplex uptake, the kinetics of rescue, intracellular trafficking, and the effects of lysosomotropic agents. We found the facilitating role of excess chitosan to be downstream of cellular uptake. Live-cell confocal quantification of intracellular trafficking revealed prolonged colocalisation of low N:P polyplexes within lysosomes, compared to shorter residence times for both rescued or N:P 5 samples, followed by observation of free pDNA in the cytosol. These data demonstrate that excess polycation mediates enhanced transfection efficiency by promoting the release of polyplexes from the endo-lysosomal vesicles, revealing a critical intracellular barrier overcome by excess polycation and suggesting possible avenues for further optimisation of polymer-based gene delivery., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

It is here described the comparison of extraction efficiency of some solutions (acetate buffer, deionized water, diluted HNO(3) and EDTA) frequently adopted in literature for evaluating the elemental solubility in airborne particulate matter. This comparison was performed considering the distribution of As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Na, Ni, Pb, S, Si, Sb, Sn, Sr, Ti, V, Zn between the extractable and mineralized residual fractions on the NIST 1648 certified material, PM(10) real samples and size-segregated samples, collected by a 13-stage impactor. The extracting solutions were evaluated by comparing extractive efficiencies and robustness towards some factors, such as acidity and concentration of complexing species, that have great environmental variability and that could be able to modify the extractive efficiency. Furthermore, extraction methods application to size-segregated samples allowed estimating the selectivity of extracting solutions towards dimensionally characterized emission sources, as dusts originated from abrasion and road dust re-suspension. On the basis of the obtained results, it was possible to define the main advantages and disadvantages resulting from the use of different extracting solutions, necessary to make possible the comparison of environmental studies carried out in different extractive conditions and to start up a proper study for harmonizing extracting procedures., (Copyright 2010 Elsevier B.V. All rights reserved.)

Objective: Nonlymphoma-associated bcl-2/IgH rearrangements (NLABRs) are frequently amplified by PCR in blood of lymphoma-free subjects (LFS), but the temporal kinetics and phenotypic nature of NLABR-positive cells are unknown. To address these issues we prospectively monitored a panel of NLABR-positive LFS., Methods: LFS have been studied by nested PCR, real-time PCR, and DNA sequencing. Cell selection studies were also performed to define the nature of NLABR-bearing clones., Results: Of 125 donors, 16 (12.8%) were found to be bcl-2/IgH positive and were monitored at least every 6 months for a median time of 22 months (range 6-50). In half of the subjects the same NLABR detected initially was again reamplified at follow-up thrice or more. In 5, the same NLABR was constantly amplified in every follow-up sample. With a median follow-up of 22 months (range 9-50), no stable disappearance of a recurrent clone has been so far recorded. Real-time PCR indicated that persistent NLABR-positive clones are stable over time in the same subject. Cell separation studies indicate that NLABRs belong to CD19+, CD5-, CD23-, CD10+/- cells., Conclusions: Our results indicate that NLABR-positive clones are persistent populations phenotypically related to follicular lymphoma (FL). This suggests the existence of a FL-related clonal expansion of undetermined significance, which might be either a premalignant or a nonmalignant counterpart of FL.

Molecular remission (MR) is associated with improved outcome in mantle cell lymphoma (MCL). If MR is not achieved, patients are at high risk of relapse. We retrospectively describe the molecular and clinical follow-ups of 4 patients with molecular relapses (M-rels) who were treated with rituximab. The 4 patients received rituximab-supplemented, high-dose sequential chemotherapy and autologous stem cell transplantation as induction treatment and achieved clinical remission and MR. M-rel was defined as polymerase chain reaction (PCR) positivity in 2 consecutive samples in the absence of clinical relapse. M-rels occurred at 3, 6, 39, and 52 months and were always confirmed by direct sequencing of the clonal rearrangement. Minimal residual disease was monitored by qualitative and real-time quantitative PCR. All patients received 4 courses of rituximab, with 2 additional infusions if PCR positivity remained. After 4-6 courses of rituximab, all patients re-entered MR. No clinical relapses were recorded at 3, 6, 18, and 62 months from treatment, although 1 patient had a second M-rel that was sensitive to rituximab. Our results indicate that rituximab is active against residual MCL cells and suggest that molecularly tailored maintenance therapy needs to be investigated in clinical trials.

Cyclooxygenase 2 (COX-2) is an inflammation-associated enzyme involved in the pathogenesis of many solid tumors, but little is known about its presence and role in hematologic neoplasms. Multiple myeloma (MM) is known to involve a deregulated cytokine network with secretion of inflammatory mediators. We thus decided to investigate the involvement of COX-2 in this neoplasm. Western blotting (WB) was used to evaluate 142 bone marrow (BM) specimens, including MM and monoclonal gammopathy of undetermined significance (MGUS). Selected cases under-went further evaluation by WB on purified CD138(+) cells, immunohistochemistry (IC), and real-time polymerase chain reaction (PCR) for mRNA expression. COX-2 was expressed in 11% (2 of 18) of MGUS specimens, 31% (29 of 94) of MM at diagnosis, and 47% (14 of 30) of MM with relapsed/refractory disease. COX-2 positivity was associated with a poor outcome in terms of progression-free (18 vs 36 months; P < .001) and overall survival (28 vs 52 months; P < .05). Real-time PCR showed COX-2 mRNA overexpression. IC and cell separation studies demonstrated COX-2 expression to be restricted to malignant plasma cells. This is the first report of the presence and prognostic role of COX-2 expression in MM. Future studies will assess COX-2 involvement in other hematologic tumors and its potential use as a therapeutic or chemo-preventive target in onco-hematology.

The evaluation of minimal residual disease (MRD) is critical in the evaluation of treatments aimed at maximal cytoreduction in multiple myeloma (MM). Qualitative evaluation of MRD now has a 10-yr-long history, but it remains a relatively sophisticated procedure. More recently, real-time quantitative approaches have also been developed. These approaches allow a very effective monitoring of disease but introduce additional complexity and costs to the procedure. This chapter describes how we currently perform real-time polymerase chain reaction (PCR) in MM. Compared to the first description of the assay in June 2000, significant improvements have been made. Although real-time PCR is the main focus of the chapter, most of the information suitable for a proper setup of a qualitative approach is also provided.

In this study we investigated telomere restriction fragment (TRF) length in a panel of mature B-cell lymphoproliferative disorders (MBCLDs) and correlated this parameter with histology and histopathogenesis in relation to the germinal center (GC). We assessed 123 MBCLD samples containing 80% or more tumor cells. TRF length was evaluated by Southern blot analysis using a chemiluminescence-based assay. GC status was assessed through screening for stable and ongoing somatic mutations within the immunoglobulin heavy-chain genes. Median TRF length was 6170 bp (range, 1896-11 200 bp) and did not correlate with patient age or sex. TRF length was greater in diffuse large cell lymphoma, Burkitt lymphoma, and follicular lymphoma (medians: 7789 bp, 9471 bp, and 7383 bp, respectively) than in mantle cell lymphoma and chronic lymphocytic leukemia (medians: 3582 bp and 4346 bp, respectively). GC-derived MBCLDs had the longest telomeres, whereas those arising from GC-inexperienced cells had the shortest (P < 10(-9)). We conclude that (1) TRF length in MBCLD is highly heterogeneous; (2) GC-derived tumors have long telomeres, suggesting that minimal telomere erosion occurs during GC-derived lymphomagenesis; and (3) the short TRF lengths of GC-inexperienced MBCLDs indicates that these neoplasms are good candidates for treatment with telomerase inhibitors, a class of molecules currently the subject of extensive preclinical evaluation.

Purpose: To evaluate the prognostic relevance of molecular monitoring of minimal residual disease in indolent lymphomas receiving high-dose sequential chemotherapy and autografting., Patients, Materials, and Methods: A polymerase chain reaction- (PCR-)based strategy was used to evaluate the presence of residual tumor cells in a panel of 70 indolent lymphoma patients: 40 with follicular (FCL), 14 with small lymphocytic (SLL), and 16 with mantle-cell (MCL) lymphomas. They were treated either with first-line (n = 61) or second-line (n = 9) therapy with an intensified high-dose chemotherapy program followed by peripheral-blood progenitor cells autografting. The Bcl-1, Bcl-2, and immunoglobulin gene rearrangements were used as lymphoma-specific markers. Overall, a molecular marker was obtained from the diagnostic tissue in 60 of 70 patients (86%). Results The collection of PCR-negative cells and the achievement of posttransplantation molecular remission (MR) were common in patients with FCL subtype (54% and 70%, respectively), whereas they were not frequent among SLL and MCL (25% and 12.5%, respectively) patients. With a median molecular follow-up of 75 months, an 88% incidence of relapse was observed among patients never attaining MR. In contrast, relapse incidence was only 8% among patients attaining a durable MR (P <.005). At present, 26 patients (20 with FCL and six with non-FCL) are long-term survivors in absence of clinical and molecular disease., Conclusion: Our results indicate that among indolent lymphomas, FCL and non-FCL subtypes show a significantly different behavior in terms of MR achievement, and MR after intensive chemotherapy and autografting is predictive for a prolonged disease-free survival, whereas persistent PCR positivity is associated with a high risk of relapse.

Objective: The aim of this study was to evaluate whether reappearance of polymerase chain reaction (PCR) positivity for the Bcl-2/IgH translocation following a phase of molecular remission in autografted follicular lymphoma (FL) patients is always associated with reappearance of the original neoplastic clone., Patients and Methods: The molecular follow-up of 119 autografted Bcl-2/IgH positive patients was evaluated by nested PCR. In case of molecular recurrence, direct sequencing of involved rearrangements has been performed both at diagnosis and at the time of recurrence. The two sequences then were compared in terms of breakpoints, N insertions, and JH usage., Results: Seventy-five patients achieving molecular remission were identified in our patient sample (63%). Of these patients, eight (10.6%) experienced molecular recurrence. Direct sequencing of the Bcl-2/IgH translocation performed at diagnosis and recurrence showed identical rearrangements in six subjects and unrelated rearrangements in two. As opposed to most true molecular relapses, unrelated rearrangements always occurred several years after transplantation. To date, the two subjects carrying unrelated rearrangements show no signs of active lymphoproliferative disease., Conclusions: This report is the first evidence that Bcl-2/IgH rearrangements unrelated to the original tumor clone can lead to false-positive results during the molecular follow-up of autografted FL patients. Based on these results, we recommend confirmation by direct sequencing, at least for patients experiencing molecular relapse 2 or more years after the end of treatment. This will be particularly important for patients enrolled in clinical trials that schedule additional treatment in case of molecular evidence of persistent disease activity.

Purpose: To assess whether nonneoplastic Bcl-2/IgH rearrangements act as a confounding factor in the setting of minimal residual disease analysis by evaluating their incidence in a panel of lymphoma-free subjects, including cancer-free donors and chemotherapy-naive and chemotherapy-treated cancer patients., Patients and Methods: A total of 501 nonlymphoma subjects have been assessed: 258 cancer-free patients and 243 patients with malignancies other than lymphoma, 112 of whom were chemotherapy-naive. Patients were primarily assessed by nested polymerase chain reaction (PCR), followed by real-time quantitative PCR if they scored positive. In addition, six initially PCR-positive cancer-free donors were prospectively reassessed by qualitative and quantitative PCR after 30 and 60 days., Results: The overall incidence of Bcl-2/IgH positivity was 9.6%, with a median number of 11 rearrangements per 1,000,000 diploid genomes (range, 0 to 2,845 rearrangements), as assessed by real-time PCR. The incidence was similar in healthy subjects and cancer patients at diagnosis (12% and 12.5%; P = not significant). In contrast, the incidence of this translocation was only 2.3% in chemotherapy-treated patients (P <.001). In addition, three initially PCR-positive cancer-free donors showed persistence of their rearrangements when assessed after 30 and 60 days., Conclusion: The low incidence of nonneoplastic Bcl-2/IgH rearrangements following chemotherapy provides further evidence of the prognostic role of persistent PCR-positivity in the posttreatment molecular follow-up of follicular lymphoma patients.

Single-center experiences have shown that intensified treatments with autologous transplantation are a promising therapeutic strategy for patients with high-risk follicle-center lymphoma (FCL) at diagnosis, whereas data from prospective multicenter trials are still lacking. This paper describes the results of a prospective multicenter study of an intensified purging-free high-dose sequential (i-HDS) chemotherapy schedule with peripheral blood progenitor cell (PBPC) autografting. The main feature of this program is harvesting stem cells after intensified chemotherapeutic debulking, with no ex vivo manipulation of PBPCs. Ninety-two previously untreated patients aged 60 or younger with advanced-stage FCL were enrolled by 20 Italian centers and evaluated on an intention-to-treat basis. i-HDS proved feasible with limited toxicity (87% patients completed the planned treatment schedule). i-HDS led to a complete remission rate of 88%. The projected overall survival and disease-free survival (DFS) were, respectively, 84% and 67% at 4 years. Centralized molecular analysis showed that polymerase chain reaction-negative harvests could be collected in 47% of cases. Following autograft, 65% of molecularly evaluable patients achieved clinical and molecular remission. The projected DFS at 4 years of this subgroup is 85%. This result emphasizes the importance of achieving maximal tumor reduction in these patients. In conclusion, our data show that highly effective intensified treatments can now be routinely offered to young patients with poor-risk FCL even at small institutions, with no need for sophisticated and expensive cell manipulation procedures.

Objective: Autologous transplantation of bone marrow (BM) and peripheral blood progenitor cells (PBPC) is commonly used for treatment of multiple myeloma (MM). Although both stem cell sources harbor residual clonal cells, a quantitative evaluation of their level of tumor contamination (LTC) still needs to be performed through highly accurate and reproducible approaches. In this study, we used a validated real-time polymerase chain reaction (PCR) strategy to evaluate LTC of BM and PBPC samples obtained from MM patients., Materials and Methods: The patients underwent two different mobilization courses (defined as early or late course) following two cycles of cyclophosphamide 5 g/m(2). LTC was evaluated by measuring the number of clonal immunoglobulin heavy-chain rearrangements followed by normalization of samples using the GAPDH gene., Results: Overall, 26 PBPC and 12 BM samples were analyzed. Main results are as follows. 1) PBPC harvests are less contaminated than BM samples taken immediately after each mobilization course (median difference 2.68 logs; range 1.7 to 4.6) (p < 0.0001). 2) LTC of PBPC harvests has only minimal variation among different leukaphereses performed during the same mobilization course (median difference 0.45 logs; range 0.22 to 1.2). 3) No difference was observed among PBPC and BM samples obtained after the late mobilization course as compared to the early mobilization course (median reduction 0.21 logs; range -0.39 to 1.3) (p = 0.84). 4) In PBPC but not in BM samples, there is a clear overestimation of the percentage of plasma cells when flow cytometric evaluation of CD38(bright) cells is compared to real-time PCR results. This suggests that in PBPC, most CD38(bright) cells do not belong to the neoplastic clone., Conclusions: Real-time PCR using the IgH rearrangement proved an effective tool for monitoring LTC in stem cell harvests from MM patients. The smaller LTC of PBPC harvests supports the role of PBPC as stem cell rescue for MM patients compared to BM cells.