CD8+ NKs as a potential biomarker of complete response and survival with lenalidomide plus R-GDP in the R2-GDP-GOTEL trial in recurrent/refractory diffuse large B cell lymphoma.

Background: Diffuse large B cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma worldwide. DLBCL is an aggressive disease that can be cured with upfront standard chemoimmunotherapy schedules. However, in approximately 35-40% of the patients DLBCL relapses, and therefore, especially in this setting, the search for new prognostic and predictive biomarkers is an urgent need. Natural killer (NK) are effector cells characterized by playing an important role in antitumor immunity due to their cytotoxic capacity and a subset of circulating NK that express CD8 have a higher cytotoxic function. In this substudy of the R2-GDP-GOTEL trial, we have evaluated blood CD8+ NK cells as a predictor of treatment response and survival in relapsed/refractory (R/R) DLBCL patients. Methods: 78 patients received the R2-GDP schedule in the phase II trial. Blood samples were analyzed by flow cytometry. Statistical analyses were carried out in order to identify the prognostic potential of CD8+ NKs at baseline in R/R DLBCL patients. Results: Our results showed that the number of circulating CD8+ NKs in R/R DLBCL patients were lower than in healthy donors, and it did not change during and after treatment. Nevertheless, the level of blood CD8+ NKs at baseline was associated with complete responses in patients with R/R DLBCL. In addition, we also demonstrated that CD8+ NKs levels have potential prognostic value in terms of overall survival in R/R DLBCL patients. Conclusion: CD8+ NKs represent a new biomarker with prediction and prognosis potential to be considered in the clinical management of patients with R/R DLBCL. Clinical trial registration: https://www.clinicaltrialsregister.eu/ctr-search/search?query=2014-001620-29 EudraCT, ID:2014-001620-29.

Lenalidomide plus R-GDP (R2-GDP) in Relapsed/Refractory Diffuse Large B-Cell Lymphoma: Final Results of the R2-GDP-GOTEL Trial and Immune Biomarker Subanalysis.

PURPOSE: New therapeutic options are needed in relapsed/refractory diffuse large B-cell lymphoma (R/R DLBCL). Lenalidomide-based schedules can reverse rituximab refractoriness in lymphoma. PATIENTS AND METHODS: In the phase II R2-GDP trial, 78 patients unsuitable for autologous stem cell transplant received treatment with the following schedule: lenalidomide 10 mg Days (D)1-14, rituximab 375 mg/m2 D1, cisplatin 60 mg/m2 D1, gemcitabine 750 mg/m2 D1 and D8, and dexamethasone 20 mg D1-3, up to 6 cycles (induction phase), followed by lenalidomide 10 mg (or last lenalidomide dose received) D1-21 every 28 days (maintenance phase). Primary endpoint was overall response rate (ORR). Secondary endpoints included progression-free survival (PFS), overall survival (OS), safety, and monitorization of key circulating immune biomarkers (EU Clinical Trials Register number: EudraCT 2014-001620-29). RESULTS: After a median follow-up of 37 months, ORR was 60.2% [37.1% complete responses (CR) and 23.1% partial responses (PR)]. Median OS was 12 months (47 vs. 6 months in CR vs. no CR); median PFS was 9 months (34 vs. 5 months in CR vs. no CR). In the primary refractory population, ORR was 45.5% (21.2% CR and 24.3% PR). Most common grade 3-4 adverse events were thrombocytopenia (60.2%), neutropenia (60.2%), anemia (26.9%), infections (15.3%), and febrile neutropenia (14.1%). Complete responses were associated with a sharp decrease in circulating myeloid-derived suppressor cells and regulatory T cells. CONCLUSIONS: R2-GDP schedule is feasible and highly active in R/R DLBCL, including the primary refractory population. Immune biomarkers showed differences in responders versus progressors.

Long-term outcome of cementless total hip arthroplasty with threaded Tropic® acetabular cup and Corail® femoral stem.

This study investigates the clinical and radiological results of a tapered femoral stem (Corail®) and uncemented threaded acetabular cups (Tropic®) and in addition an analysis of the complications and retrieved implants was conducted. Between January 1990 and September 1998, 301 total hips arthroplasties in 268 patients were implanted. 78 patients (87 hips) had died and 9 patients (12 hips) had been lost to follow-up, leaving at the time of the latest follow-up 180 patients (202 hips). The mean age at surgery was 56,1 (27-75) years. Of the 154 unrevised hips, the median Harris and Merle d'Aubigne and Postel hip scores were 83,3 points and 15,3 points respectively at the latest follow-up. The median follow-up time was 16.9 years (10,4-25). No femoral component was revised for aseptic loosening ; osteolysis was observed in the 9,5% of the implants (13 stems). 48 hips (23%) were revised and 27 cups (56,2%) needed revision surgery because of massive polyethylene wear. Pelvic osteolysis was found out in 80 cups (58,8%). 87 hips (63,9%) showed signs of a progressive wear of the liner, more than 2mm in 48 hips. Kaplan-Meier survivorship analysis at 15 years follow-up was 81.2% with revision for any reason (85.8% for mechanical or radiographic loosening). High rates of polyethylene wear and the high prevalence of pelvic osteolysis are serious matters in these types of implants with high rates of revision at 15 years follow-up so we decided to abandon the concept of a threaded cup design in favor of a press-fit acetabular cup.

Circulating myeloid-derived suppressor cells and regulatory T cells as immunological biomarkers in refractory/relapsed diffuse large B-cell lymphoma: translational results from the R2-GDP-GOTEL trial.

BACKGROUND: The search for immunological markers with ability of predicting clinical outcome is a priority in lymphomas, and in cancer in general. It is well known that some immunomodulatory cells, such as myeloid derived suppressor cells (MDSCs) or regulatory T cells (Tregs), are recruited by tumors, jeopardizing antitumor immunosurveillance. In this work, we have studied blood levels of these immunosuppressive cells in patients with relapsed/refractory diffuse large B-cell lymphoma (R/R DLBCL), prior to and along the course of the experimental rituximab, gemcitabine, dexamethasone, and cisplatin (R2-GDP) schedule, as a translational substudy of the R2-GDP-GOTEL trial (EudraCT Number: 2014-001620-29), which included lenalidomide as an immunomodulator. METHODS: Blood samples were taken before treatment, at cycle 3 and end of induction. Samples were analyzed by flow cytometry. Non-parametric tests were used. Mann-Whitney U test was used to compare basal cells distributions, and Wilcoxon test was considered to compare cells distribution at different times. Spearman test was performed to measure the degree of association between cell populations. RESULTS: In this study, MDSC and Treg circulating concentration was found increased in all patients compared with a healthy control group and decreased after treatment only in patients with longest overall survival (>24 months), reaching the levels of the healthy group. Likewise, the number of inhibited T lymphocytes expressing Programmed Death-1 (PD-1) were increased in peripheral blood from patients and decreased on the treatment, whereas activated T lymphocytes increased after therapy in those with better overall survival. CONCLUSIONS: In conclusion, blood concentration of MDSCs and Treg cells may be good prognostic markers for overall survival after 2 years in R/R DLBCL. These results point to a possible role of these elements in the immunosuppression of these patients, as assessed by the circulating activated and inhibited T lymphocytes, and therefore, they may be considered as therapeutic targets in DLBCL.

Osteosarcoma of the Trapezium.

Osteogenic sarcoma is a malignant tumor that rarely affects the hand. When it does, it most often involves the phalanges or metacarpal heads. We present the case of a 51-year-old woman with a low-grade osteosarcoma affecting the trapezium bone of her left hand. A total trapeziectomy with partial removal of the first metatarsal, scaphoid, trapezoid, and capitate bones was performed, and no adjuvant therapy was administered. Six years after the intervention, the patient is disease-free, with excellent functionality and yearly imaging tests showing no signs of recurrence.

Microvesicles from Human Adipose Tissue-Derived Mesenchymal Stem Cells as a New Protective Strategy in Osteoarthritic Chondrocytes.

BACKGROUND/AIMS: Chronic inflammation contributes to cartilage degeneration during the progression of osteoarthritis (OA). Adipose tissue-derived mesenchymal stem cells (AD-MSC) show great potential to treat inflammatory and degradative processes in OA and have demonstrated paracrine effects in chondrocytes. In the present work, we have isolated and characterized the extracellular vesicles from human AD-MSC to investigate their role in the chondroprotective actions of these cells. METHODS: AD-MSC were isolated by collagenase treatment from adipose tissue from healthy individuals subjected to abdominal lipectomy surgery. Microvesicles and exosomes were obtained from conditioned medium by filtration and differential centrifugation. Chondrocytes from OA patients were used in primary culture and stimulated with 10 ng/ml interleukin(IL)-1ß in the presence or absence of AD-MSC microvesicles, exosomes or conditioned medium. Protein expression was investigated by ELISA and immunofluorescence, transcription factor-DNA binding by ELISA, gene expression by real-time PCR, prostaglandin E2 (PGE2) by radioimmunoassay, and matrix metalloproteinase (MMP) activity and nitric oxide (NO) production by fluorometry. RESULTS: In OA chondrocytes stimulated with IL-1ß, microvesicles and exosomes reduced the production of inflammatory mediators tumor necrosis factor-α, IL-6, PGE2 and NO. The downregulation of cyclooxygenase-2 and microsomal prostaglandin E synthase-1 would lead to the decreased PGE2 production while the effect on NO could depend on the reduction of inducible nitric oxide synthase expression. Treatment of OA chondrocytes with extracellular vesicles also decreased the release of MMP activity and MMP-13 expression whereas the production of the anti-inflammatory cytokine IL-10 and the expression of collagen II were significantly enhanced. The reduction of inflammatory and catabolic mediators could be the consequence of a lower activation of nuclear factor-κB and activator protein-1. The upregulation of annexin A1 specially in MV may contribute to the anti-inflammatory and chondroprotective effects of AD-MSC. CONCLUSIONS: Our data support the interest of AD-MSC extracellular vesicles to develop new therapeutic approaches in joint conditions.

Revision of infected total knee arthroplasty: two-stage reimplantation using an antibiotic-impregnated static spacer.

BACKGROUND: A two-stage revision remains as the "gold standard" treatment for chronically infected total knee arthroplasties. METHODS: Forty-five septic knee prostheses were revised with a minimum follow-up of 5 years. Static antibiotic-impregnated cement spacers were used in all cases. Intravenous antibiotics according to sensitivity test of the culture were applied during patients' hospital stay. Oral antibiotics were given for another 5 weeks. Second-stage surgery was undertaken after control of infection with normal erythrocyte sedimentation rate and C-reactive protein values. Extensile techniques were used if needed and metallic augments were employed for bone loss in 32 femoral and 29 tibial revisions. RESULTS: The average interval between the first-stage resection and reimplantation was 4.4 months. Significant improvement was obtained with respect to visual analog scale pain and clinical and functional scores, and infection was eradicated in 95.6% of cases following a two-stage revision total knee arthroplasty. Radiographic evaluation showed suitable alignment without signs of mechanical loosening. CONCLUSIONS: This technique is a reasonable procedure to eradicate chronic infection in knee arthroplasty and provides proper functional and clinical results. However, it sometimes requires extensile surgical approaches that could imply arduous surgeries. Metallic augments with cementless stems available in most of the knee revision systems are a suitable alternative to handle bone deficiencies, avoiding the use of bone allografts with its complications.

Darbepoetin alfa administration in patients with non-Hodgkin lymphoma and chemotherapy-induced anemia receiving (±R) CHOP.

IMPACT NHL was a multicenter, observational study in adults with non-Hodgkin lymphoma receiving CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisolone) chemotherapy with or without rituximab. Erythropoietin-stimulating agent treatment was given according to routine clinical practice and physician preference. In a subanalysis, outcomes were evaluated in 207 patients who received darbepoetin alfa (DA). The most common reason (81%) for initiating DA was low/declining hemoglobin (Hb) concentration. Mean (±standard deviation) duration of DA exposure was 8.8 ± 6.9 weeks (mean number of doses, 5.1 ± 4.6). Overall, 23% of patients had chemotherapy and DA treatment synchronized more than 75% of the time. At the time of DA initiation, 67% of patients had Hb concentrations in the guideline-recommended range (9-11 g/dl). Of 89 patients with Hb concentrations <10 g/dl at DA initiation and still receiving DA 5 weeks later, 92% (Kaplan-Meier) achieved Hb concentrations 10-12 g/dl between week 5 and at the end of treatment.

Impact of age group on febrile neutropenia risk assessment and management in patients with diffuse large B-cell lymphoma treated with R-CHOP regimens.

UNLABELLED: Improving the management of elderly patients with lymphoma is of increasing relevance. One thousand one hundred thirteen patients with diffuse large B-cell lymphoma (DLBCL) received rituximab (R)-CHOP (cyclophosphamide, doxorubicin [hydroxydaunorubicin], vincristine [Oncovin], and prednisone) in an observational study. Both older and younger patients failed to receive growth factor support in accordance with international guidelines; patients 65 years and older were more susceptible to febrile neutropenia (FN) and its consequences. Better application of guidelines could reduce rates of FN and improve outcomes. BACKGROUND: The incidence of diffuse large B-cell lymphoma (DLBCL) is increasing in the elderly population, which is a more challenging population to treat because of comorbidities and enhanced sensitivity to chemotherapy toxicities. This analysis evaluated the impact of age group on assessment of febrile neutropenia (FN) risk, supportive care management, and chemotherapy delivery. METHODS: The IMPACT non-Hodgkin lymphoma (NHL) trial was an observational study conducted in Europe and Australia. This analysis included 1113 patients with DLBCL treated with rituximab (R)-CHOP (cyclophosphamide, doxorubicin [hydroxydaunorubicin], vincristine [Oncovin], and prednisone) every 14 days (n = 409) or every 21 days (n = 704). Outcomes were reported for ages < 65 years and ≥ 65 years. The primary outcome in this analysis was the proportion of patients assessed by investigators as having an overall high (≥ 20%) FN risk who received granulocyte colony-stimulating factor (G-CSF) primary prophylaxis. RESULTS: For R-CHOP-14, investigators assessed 78% of younger patients and 80% of older patients with ≥ 20% risk of FN, although 14% of younger and 19% of older high-risk patients did not receive G-CSF primary prophylaxis. For R-CHOP-21, investigators assessed 52% of younger and 71% of older patients with ≥ 20% risk of FN; however, 61% of younger and 47% of older high-risk patients did not receive G-CSF primary prophylaxis. Regardless of chemotherapy regimen, rates of FN and unplanned hospitalization were higher in older patients, and delivery of chemotherapy was poorer. CONCLUSION: Adherence to G-CSF guidelines in patients assessed with high FN risk was suboptimal in patients with DLBCL receiving R-CHOP chemotherapy, with substantial proportions of both younger and older patients receiving R-CHOP-21 failing to receive optimal G-CSF support. Better application of guidelines could reduce FN rates and improve outcomes.

Impact of febrile neutropenia on R-CHOP chemotherapy delivery and hospitalizations among patients with diffuse large B-cell lymphoma.

PURPOSE: This analysis from an observational study of clinical practice describes the impact of febrile neutropenia (FN) on chemotherapy delivery and hospitalizations. METHODS: Adults with diffuse large B-cell lymphoma (DLBCL) scheduled to receive ≥ 3 cycles of 2- or 3-weekly CHOP with rituximab (R-CHOP-14/21) were eligible. Primary outcome was incidence of FN. RESULTS: FN data were available for 409 patients receiving R-CHOP-14 and 702 patients receiving R-CHOP-21. FN incidence was R-CHOP-14, 20% (81/409) and R-CHOP-21, 19% (133/702). Rates of primary prophylaxis with granulocyte-colony stimulating factor were R-CHOP-14, 84% (345/409) and R-CHOP-21, 36% (252/702). A large number of patients experienced their first FN episode in cycle 1 (R-CHOP-14, 24/81 [30%]; R-CHOP-21, 63/133 [47%]). Multiple risk factors (≥ 2) for FN were more frequent in patients experiencing FN than in patients not experiencing FN (R-CHOP-14, 60/81 [74%] versus 179/328 [55%]; R-CHOP-21, 98/133 [74%] versus 339/569 [60%]). A similar trend was observed for unplanned hospitalizations (R-CHOP-14, 63/81 [78%] versus 68/328 [21%]; R-CHOP-21, 105/133 [79%] versus 100/569 [18%]). Achievement of chemotherapy relative dose intensity ≥ 90% was lower among patients experiencing FN than in patients not experiencing FN (R-CHOP-14, 30/81 [37%] versus 234/328 [71%]; R-CHOP-21, 83/133 [62%] versus 434/569 [76%]). CONCLUSIONS: In patients with DLBCL treated with R-CHOP-14 or R-CHOP-21, patients with an event of FN were more likely to experience suboptimal chemotherapy delivery and increased incidence of unplanned hospitalizations than those without FN. FN-related hospitalizations are likely to impact chemotherapy delivery and to incur substantial costs.

Individuals infected with JC polyomavirus do not present detectable JC virus DNA in oropharyngeal fluids.

JC virus (JCV) is ubiquitous in the human population. Primary infection normally occurs during childhood and is followed by a lifelong persistent infection. The main mode of transmission remains unknown. Several authors have hypothesized that JCV transmission occurs through the respiratory route, and that respiratory secretions could represent a possible source of viral particles. The present study intended to evaluate oropharyngeal fluids from patients infected with JCV, in order to ascertain if respiratory secretions could indeed constitute a source of exposure to this polyomavirus. Oropharyngeal washing samples from 25 patients co-infected with JCV and human immunodeficiency virus type 1 were evaluated for the presence of JCV DNA. Regardless of the titre of antibodies or the presence of viral urinary excretion, JCV genome was not detected in oropharyngeal samples collected from any of the patients infected with JCV included in this study, which may suggest that oropharyngeal fluids are an unlikely source for JCV infection.

Seroepidemiological survey of transmissible infectious diseases in a portuguese prison establishment.

A cross-sectional study was conducted in 151 (71.6%) of 211 male inmates of a regional Portuguese prison in order to establish the seroprevalence for viral hepatitis (HAV, HBV, HCV), human immunodeficiency virus (HIV), syphilis and herpes simplex virus (HSV-1 and HSV-2) and to analyze some psychosocial and criminal characteristics. Mean age was 34 years. Anti-HAV was positive in 69.5% (n = 105) and in 34.4% (n = 52) for anti-HCV. One (0.7%) person had HBsAg and 29 (19.2%) had laboratory markers of past HBV infection. Non-immune inmates for HBV were 40.4% (n = 61). Syphilis was diagnosed in 6.0% (n = 9). The rate of HIV infection was 6.6% (n = 10; all HIV-1). The seropositivity of HSV-2 was 19.9% (n = 30) and of HSV-1 was 82.1% (n = 124). Alcohol dependence was reported by 26.5% (n = 40). Excluding tobacco and prescription medication, 73.5% (n = 111) reported drug use in prison. The most commonly used drugs were: cannabis (100%; n = 111) followed by heroin (56.7%; n = 63). Anti-HCV rate was noteworthy. The HIV infection rate (6.6%) in this regional prison is at least 13 to 22 times greater than in general population. As the inmate return to the community increases the risk of disease exposure for the general population, early detection and counseling is urgently needed for prisoners.

Seroepidemiological survey of transmissible infectious diseases in a portuguese prison establishment

A cross-sectional study was conducted in 151 (71.6 percent) of 211 male inmates of a regional Portuguese prison in order to establish the seroprevalence for viral hepatitis (HAV, HBV, HCV), human immunodeficiency virus (HIV), syphilis and herpes simplex virus (HSV-1 and HSV-2) and to analyze some psychosocial and criminal characteristics. Mean age was 34 years. Anti-HAV was positive in 69.5 percent (n = 105) and in 34.4 percent (n = 52) for anti-HCV. One (0.7 percent) person had HBsAg and 29 (19.2 percent) had laboratory markers of past HBV infection. Non-immune inmates for HBV were 40.4 percent (n = 61). Syphilis was diagnosed in 6.0 percent (n = 9). The rate of HIV infection was 6.6 percent (n = 10; all HIV-1). The seropositivity of HSV-2 was 19.9 percent (n = 30) and of HSV-1 was 82.1 percent (n = 124). Alcohol dependence was reported by 26.5 percent (n = 40). Excluding tobacco and prescription medication, 73.5 percent (n = 111) reported drug use in prison. The most commonly used drugs were: cannabis (100 percent; n = 111) followed by heroin (56.7 percent; n = 63). Anti-HCV rate was noteworthy. The HIV infection rate (6.6 percent) in this regional prison is at least 13 to 22 times greater than in general population. As the inmate return to the community increases the risk of disease exposure for the general population, early detection and counseling is urgently needed for prisoners.

Mid- to long-term outcome of cementless total hip arthroplasty in younger patients.

PURPOSE: To assess mid- to long-term outcomes of cementless primary total hip arthroplasty (THA) in younger patients. METHODS: Records of 28 women and 34 men (75 hips) aged 18 to 55 (mean, 38) years who underwent primary THA using a hydroxyapatite-coated stem and a threaded cup and had been followed up for a mean of 10 (6-15) years were reviewed. 13 of the patients had bilateral THAs. Clinical and radiographic outcomes were evaluated. RESULTS: After a minimum follow-up of 7 (range, 7-14) years, 12 (16%) of the hips were revised, of which 8 (11%) were for the cup. The causes for revision were late deep infections (n=2), aseptic loosening of the cup (n=4), and polyethylene wear (n=6). No stem was revised for aseptic loosening. Osteolysis was noted in 24 (38%) hips. Of the 63 unrevised hips, the mean Harris Hip Score was 92 (range, 45-100) and the mean D'Aubigne and Postel score was 17 (range, 9-18). The survival of the threaded cup was 88% at 10 years, using revision surgery as the end point. 27 (41%) of the hips showed signs of polyethylene wear; 15 were >2 mm. CONCLUSION: In younger patients undergoing THA, rates of polyethylene wear and pelvic osteolysis are high, and thus long-term follow-up is crucial.

Characterization of JC human polyomavirus infection in a Portuguese population.

JC virus (JCV) is ubiquitous in the human population, infecting children asymptomatically. After primary infection, JCV persists in the host throughout life and is often excreted in the urine. Two hundred thirty-four urine samples and 78 serum samples, collected from 171 healthy individuals and 63 patients infected with HIV, were used to characterize JCV infection in a Portuguese population. Using PCR, JCV DNA was detected in 38% of the urine samples. A significant difference in the excretion rate was observed between patients infected with HIV (51%) and healthy individuals (33%). The frequency of JCV viruria increased with age in healthy individuals, but not in patients infected with HIV. JCV urinary load was determined by real-time quantitative PCR and was independent of gender, age, HIV infection, and CD4+ cell count. Overall, the JCV genotype detected most commonly was 1B, followed by genotypes 2B and 4. The detection and quantitation of JCV-specific antibodies were performed in serum samples by an established enzyme immunoassay (EIA). Antibodies to JCV were observed in 91% of the patients tested, irrespective of HIV infection. A positive correlation between JCV urinary load and antibody titers was demonstrated. The present study provides the first characterization of seroprevalence and urinary excretion of JCV in a Portuguese population and revealed similar results to those observed in other European countries. A comparison between healthy individuals and patients infected with HIV, despite identical values of seroprevalence, showed some differences in the pattern of urinary excretion. J. Med. Virol. 82:494-504, 2010. (c) 2010 Wiley-Liss, Inc.

Passive hip laxity in Estrela Mountain Dog--distraction index, heritability and breeding values.

Two hundred and fifteen Estrela Mountain Dogs (EMD) were examined using the PennHIP method between 2002 and 2006. Passive hip laxity (PHL) was estimated calculating the distraction index (DI). Pedigree information was obtained from the Portuguese Kennel Club. The heritability and breeding values were estimated using the linear Animal Model. The DI, using as reference the worst joint of each animal, ranged from 0.15 to 1.12 (0.55 +/- 0.19). The PHL was equal in right and left sides, and was higher in females than in males (P > 0.05 and P <0.01 in the t-test, respectively). The estimated heritability was 0.83 +/- 0.11. The mean breeding values for PHL were stable in EMD born between 1991 and 2003, and showed an improvement in 2004 and 2005. The data confirm high PHL in breeds with high prevalence and severity of canine hip dysplasia. The high heritability indicates that the DI could be reduced in the breed if PHL were used as a selection criterion. The early favourable evolution of DI breeding values could be related with the increase of voluntary radiographic hip screening in the last years, and the subsequent introduction of hip quality as a breeding criterion.

Ramón Alfaro y la anestesia mexicana / Ramón Alfaro and the Mexican anesthesia

Recientes investigaciones han permitido sacar a la luz los artículos que el doctor Ramón Alfaro publicó entre 1840 y 1868. A través de éstos es posible delinear al doctor Alfaro como un perseverante profesional de la medicina, imbuido de un particular espíritu científico que lo condujo a desarrollar cuidadosos trabajos de investigación en relación con los problemas médicos de la época. Uno de los primeros trabajos se refiere de manera específica al cloroformo. Así, el doctor Alfaro realizó una amplia revisión sobre el tema, comunicó la primera casuística de pacientes quirúrgicos sometidos a la anestesia inhalada y sentó las bases para una práctica anestésica más racional y segura.

Recent historical inquiries have shed light on a series of articles published between 1840 and 1868 by Ramón Alfaro M.D. His publications reflect the work of a persevering medical researcher who carried out studies of health conditions endemic at the time. One of his first publications is devoted to chloroform. In it, Dr. Alfaro made a careful assessment on the use of anesthesia, published the first statistical study of the impact of inhalational anesthesia in a number of surgical procedures, and was able to establish the foundations for more rational and safe anesthetic procedures.