Syngeneic transplantation with peripheral blood mononuclear cells collected after the administration of recombinant human granulocyte colony-stimulating factor

Five syngeneic transplants were performed in four patients following myeloablative therapy using unmodified peripheral blood mononuclear cells (PBMCs) collected after the administration of recombinant human granulocyte colony stimulating factor (rhG-CSF) to normal donors. The only toxicity experienced by the four normal donors was bone pain. Four patients received two collections of PBMCs, and a second transplant was performed in one patient with one collection. The patients received a median of 20.53 x 10(8) total nucleated cells/kg (range 20 to 25.5), 11.3 x 10(8) total mononuclear cells/kg (range 6.52 to 17.2), 113.1 x 10(4)/kg CFU-GM (range 46.7 to 211.8) and 9.6 x 10(6) CD34+ cells/kg (range 1.6 to 12.6) Post-transplant growth factors were not administered. The median time to an absolute neutrophil count greater than 0.5 x 10(9)/L was 14 days (range 10 to 18). The median time to platelet transfusion independence was 11 days (range 10 to 13). Two patients had the number of CD3+ T lymphocytes determined in the pheresis product. An average of 3.04 x 10(10) CD3+ cells were collected per pheresis. This represents an approximate 1 log increase over the number of T lymphocytes in a typical bone marrow transplant. Rh-GCSF can be used to mobilize peripheral blood progenitor cells from normal donors with minimal toxicity. Studies of allogeneic transplants using PBMCs collected after rhG-CSF administration to determine permanent grafting ability and the incidence and severity of graft-versus-host disease are warranted.     

Infection with Leptospira kirschneri Serovar Mozdok: First Report from the Southern Hemisphere

Leptospirosis is a global zoonosis caused by pathogenic Leptospira spp. In this study, we characterized two Leptospira kirschneri serogroup Pomona serovar Mozdok isolates, one obtained from a dog and the other from a patient with severe leptospirosis, 4 years later. Histopathological analysis showed that both isolates caused severe tissue damage when used to infect hamsters. While L. kirschneri serogroup Pomona serovar Mozdok is endemic in animals in Europe, there is only one report of human leptospirosis in the literature. Although strains belonging to L. kirschneri serogroup Pomona have been identified in cases of human leptospirosis in Europe, serovar Mozdok has not yet been implicated. The 4-year interval between isolations and the fact that this is the first report of serovar Mozdok as the causative agent of human leptospirosis in the southern hemisphere, demonstrates its epidemiological importance to public health. Moreover, the presence of serovar Mozdok in Brazil has the potential to affect vaccine and diagnostic test development.Leptospirosis is a reemerging zoonotic disease, and the global burden is showing an upward trend. The original estimates in 1999¹ predicted some 500,000 annual cases compared with the latest prediction of 873,000 cases and 49,000 mortalities per year, a 74.6% increase over 15 years.² Accurate laboratory diagnosis continues to be a limiting factor, meaning that the true global burden of leptospirosis is likely to be much higher.³ In Latin America, the prevalence of severe leptospirosis is high (10,000 cases a year) due to the tropical climate and lack of appropriate sanitation.³ Although the city of Pelotas has a subtropical climate, > 50 cases of human leptospirosis per 100,000 inhabitants are reported each year, one of the highest rates in southern Brazil.⁴ The infection rate in Pelotas is higher than the Brazilian average for the same period (3.5/100,000) and other regions with similar climatic conditions (> 10/100,000).⁵At present, there are 10 pathogenic Leptospira spp. classified into > 260 serovars⁶ and Leptospira interrogans, Leptospira borgpetersenii, and Leptospira kirschneri are most commonly associated with human leptospirosis.⁷ In Brazil, L. interrogans serogroup Icterohaemorrhagiae serovars Icterohaemorrhagiae and Copenhageni are the main cause of urban leptospirosis and have been widely studied,³ whereas rural leptospirosis and the associated serovars have been largely neglected. To the best of our knowledge, L. kirschneri serogroup Pomona serovar Mozdok has only been implicated in a case of human leptospirosis in Cuba.⁸ Serovar Mozdok is endemic to Croatia where it is prevalent in wild rodents. Human leptospirosis caused by serogroup Pomona is common in that region and while serovar Mozdok has not been implicated in any human cases,⁹ it is a causative agent of canine leptospirosis in Europe.¹⁰We report the isolation and characterization of two isolates of serovar Mozdok recovered from cases of canine and human leptospirosis in Pelotas, southern Brazil. The canine strain was isolated in 2009 during a municipal dog neutering campaign. Urine samples were aseptically collected from the bladder during ovarian hysterectomy, via aspiration using an insulin 30-G needle and syringe (BD Biosciences, Franklin Lakes, NJ). The urine was immediately inoculated into unsupplemented Ellinghausen-McCullough-Johnson-Harris (EMJH; Difco, Sparks, MD) medium (100 μL urine/5 mL EMJH), incubated for 1 hour and then subcultured into EMJH containing 10% of a commercial supplement (Difco). The dog from which the strain was isolated was asymptomatic and was released after the surgical procedure. The second isolate was obtained from the blood culture of a 56-year-old female patient from a rural area of the city. The patient presented with headache, myalgia, fever, vomiting, fatigue, sleepiness, and arthralgia and reported contact with dogs, rats, pigs, cattle, and flood water. The isolate was cultured in EMJH medium as described for the canine isolate. Both isolates were identified as L. kirschneri by means of secY gene sequencing.¹¹ Multilocus sequence typing (MLST)⁷ further characterized the isolates as L. kirschneri serogroup Pomona serovar Mozdok (ST 117). All sequencing procedures were performed using the paired-end technology on an Illumina Solexa platform (Illumina, San Diego, CA; GenBank accession numbers for sequences are shown in

Retrovirally marked CD34-enriched peripheral blood and bone marrow cells contribute to long-term engraftment after autologous transplantation

We report here on a preliminary human autologous transplantation study of retroviral gene transfer to bone marrow (BM) and peripheral blood (PB)-derived CD34-enriched cells. Eleven patients with multiple myeloma or breast cancer had cyclophosphamide and filgrastim-mobilized PB cells CD34-enriched and transduced with a retroviral marking vector containing the neomycin resistance gene, and CD34-enriched BM cells transduced with a second marking vector also containing a neomycin resistance gene. After high-dose conditioning therapy, both transduced cell populations were reinfused and patients were followed over time for the presence of the marker gene and any adverse effects related to the gene-transfer procedure. All 10 evaluable patients had the marker gene detected at the time of engraftment, and 3 of 9 patients had persistence of the marker gene for greater than 18 months posttransplantation. The marker gene was detected in multiple lineages, including granulocytes, T cells, and B cells. The source of the marking was both the transduced PB graft and the BM graft, with a suggestion of better long-term marking originating from the PB graft. The steady- state levels of marking were low, with only 1:1000 to 1:10,000 cells positive. There was no toxicity noted, and patients did not develop detectable replication-competent helper virus at any time posttransplantation. These results suggest that mobilized PB cells may be preferable to BM for gene therapy applications and that progeny of mobilized peripheral blood cells can contribute long-term to engraftment of multiple lineages.     

Reinforcement of Single Implant‐Retained Mandibular Overdenture with a Cobalt‐Chromium Framework before Implant Surgery

Previous clinical research has shown promising results in oral rehabilitation of elderly adults by a single implant‐retained mandibular overdenture; however, the high incidence of fracture in the anterior region of these overdentures is a concern for clinicians. To minimize catastrophic overdenture fracture, we propose a technique to insert an individualized metal framework in single implant‐retained overdentures prior to implant surgery.     

Evaluation of the antioxidant activities of fatty polyhydroquinolines synthesized by Hantzsch multicomponent reactions

Polyhydroquinolines (PHQs) are the unsymmetrical Hantzsch derivatives of 1,4-dihydropyridines with several biological applications. In this work, new fatty 2- and 3-substituted PHQ derivatives from different fatty acids and fatty alcohol feedstocks were synthesized at good yields via a four-component reaction (4CR). The antioxidant activities of fatty PHQs were investigated using three different antioxidant methods. The experiments showed that the compounds derived from 2-nitrobenzaldehyde and fatty palmitic (C16:0) and oleic (C18:1) chains showed better antioxidant activity. This revealed that combining the ortho NO₂ group in the aromatic ring with the insertion of fatty chains in the PHQ core contributed to the antioxidant activity. However, among all the fatty PHQs tested, the fatty 2-substituted compound derived from oleyl alcohol and 2-nitrobenzaldehyde showed the highest antioxidant activity (EC₅₀, 2.11–4.69 μM), which was similar to those of the antioxidant standards butylated hydroxytoluene (EC₅₀, 1.98–6.47 μM) and vitamin E (EC₅₀, 1.19–5.88 μM). In addition, this lipophilic compound showed higher antioxidant activity than the antihypertensive drug nifedipine (EC₅₀, 49.25–126.86 μM). These results indicate that the new fatty PHQs may find novel applications as antioxidant additives.

The insertion of a fatty long chain into polyhydroquinoline (PHQ) core contributes to antioxidant potential, the new fatty PHQs showed activities similar to commercial antioxidants.     

Handgrip Strength at Admission and Time to Discharge in Medical and Surgical Inpatients

Background and Objective: Handgrip strength is a relevant marker of functional status and is also a component of nutrition assessment. The simplicity of this measurement supports its usefulness as a tool to predict who will likely take longer to hospital discharge. The aim of this study was to quantify the association between sex‐specific handgrip strength at hospital admission and time to discharge alive. We intended to include a group of diverse diagnoses and to compare medical and surgical wards, taking into account the potential confounders’ effect of patients’ characteristics and severity of disease. Subjects and Methods: Prospective study in 2 public acute‐care general hospitals in Porto, Portugal, in 2004. Handgrip strength was evaluated using a handgrip dynamometer in a probability sample of 425 patients from medical and surgical wards. The association between baseline handgrip strength and time to discharge was evaluated using survival analysis with discharge alive as the outcome and deaths and transfers being censored. Results: In medical wards, women with high admission handgrip strength had a very short hospital stay (all had been discharged by the sixth day), and among men, patients with low handgrip strength had a particularly longer stay (approximately 50% were discharged after 15 days of hospitalization). In surgical wards, an increasing length of stay with decreasing handgrip strength quartiles was also observed in both sexes. Conclusions: Lower handgrip strength at hospital admission was associated with a longer time in the hospital, in patients of both sexes, in medical and surgical wards. Although this association was explained in part by age, height, education level, cognitive status, and disease severity, its direction remained unchanged regardless of the aforementioned factors.     

Deleterious action of azadirachtin against the mutualistic fungus of leaf-cutting ants

Leaf-cutting ants have a beneficial and obligatory relationship with the fungus that they grow. This mutualism allowed the evolutionary success of these ants. The great defoliation capacity of these insects, which often exceeds the level of tolerable economic damage, includes them as severe pests in many cultures. However, given the close relationship between these two agents of mutualism, it is expected that an impact on the fungus will reflect on the performance of the colony as a whole. Therefore, the effect of azadirachtin on the development, and the macronutrient composition of Leucoagaricus gongylophorus was evaluated. Azadirachtin reduced the final fungal mass at the end of treatment at all concentrations tested, but did not reduce the final growth area. A reduction in the amount of hyphae produced with increasing azadirachtin concentration was also observed. Regarding macronutrients, the compound did not affect their total amount in the fungus. Thus, it is observed that azadirachtin did not alter the composition of L. gongylophorus macronutrients, but inhibited its growth by reducing the number of hyphae produced. This reduction reflects directly on the amount of nutrients offered to the workers and the queen and may improve the management of these insects.