Gastrointestinal parasite assemblages from the wild rodent capybara (Hydrochoerus hydrochaeris) inhabiting a natural protected area from Argentina.

Knowledge about parasitic diseases of wildlife will help us to understand the dynamics of parasites and their effects on host populations. The capybara (Hydrochoerus hydrochaeris) is the largest living rodent in the world, and its distribution is associated with the presence of tropical and subtropical wetlands in South America. The Los Padres Lake Integral Reserve (LPLIR) is an important conservation zone in the pampean region of Argentina. One of the emblematic species found within the reserve is the capybara. The objective of this study was to determine the gastrointestinal parasites present in wild capybaras of the LPLIR and to compare different coprological methodologies. Free-ranging capybara fresh feces from 57 individuals were randomly collected from the area of LPLIR in the summer of 2022. Three different techniques were applied: spontaneous sedimentation technique (SS), INTA modified McMaster technique (MM), and Mini-FLOTAC (MF) technique. Fifty-six samples from all samples analysed (56/57, 98%) were found to be positive for gastrointestinal parasites. Two species of Strongylida, Protozoophaga obesa, Echinocoleus hydrochaeris, one unidentified nematode, one unidentified spirurid, and at least two morphotypes of Eimeria spp. oocysts were recorded. There were found significant differences in the proportion of positive samples and in richness by technique, but no significant differences were found in parasite counting. In conclusion, the choice of methodology depends on the specific objectives of the study. This is the first parasitological study of capybaras from the LPLIR and represents an exploration of parasite communities present in these wild rodents at their southernmost distribution.

From CT scanning to 3D printing technology: a new method for the preoperative planning of a transcutaneous bone-conduction hearing device.

SUMMARY: The aim of the present study was to assess the feasibility and utility of 3D printing technology in surgical planning of a transcutaneous bone-conduction hearing device (Bonebridge®) (BB), focusing on the identification of the proper location and placement of the transducer. 3D printed (3DP) models of three human cadaveric temporal bones, previously submitted to CT scan, were created with the representation of a topographic bone thickness map and the sinus pathway on the outer surface. The 3DP model was used to detect the most suitable location for the BB. A 3DP transparent mask that faithfully reproduced the surface of both the temporal bone and the 3DP model was also developed to correctly transfer the designated BB area. The accuracy of the procedure was verified by CT scan: a radiological marker was used to evaluate the degree of correspondence of the transducer site between the 3DP model and the human temporal bone. The BB positioning was successfully performed on all human temporal bones, with no difficulties in finding the proper location of the transducer. A mean error of 0.13 mm was found when the transducer site of the 3DP model was compared to that of the human temporal bone. The employment of 3D printing technology in surgical planning of BB positioning showed feasible results. Further studies will be required to evaluate its clinical applicability.

Normal variation of bone marrow B-cell precursors according to age - reference ranges for studies in myelodysplastic syndromes in Brazil.

BACKGROUND: Normal B lymphoid maturation occurs in bone marrow (BM) throughout life, but immature B-cell progenitors (BCPs) are more numerous in children than in adults. To assess the normal values according to age became important as BCPs are decreased in myelodysplastic syndromes and have been considered an important diagnostic and prognostic feature in these clonal disorders. METHODS: in a multicenter retrospective study from the Brazilian Group of Flow Cytometry we analyzed the variation of BCPs in normal BM according to age and technical peculiarities of each laboratory. We analysed of 45 BM donors and 89 cases examined for elucidation of transitory reactive cytopenias presenting a normal BM immunophenotyping. BCPs were enumerated as CD19+ /CD34+ /CD45dim /CD10+ cells (panel 1) or CD19+ /CD34+ /CD45dim cells (panel 2) among the total nucleated non-erythroid cells and as percentage of CD34+ cells. RESULTS: we included 134 cases. Panel 1 was applied in 88 cases and panel 2 was used in 46. Age range: 10 months to 89 years. In a multiple regression, % BCPs/total nucleated cells was an exponential function of age. Age explained alone 49.4% of the variance, while 'panel used' explained 1.8% and 'laboratory' explained 0.7%. Age explained only 24.9% of the variance of BCPs/CD34+ cells. CONCLUSIONS: in normal individuals, BM B-cell precursors varied mainly according to age, but were also dependent on technical peculiarities of operators and equipments. Analysis by phenotype and as percentage of total nucleated cells was more accurate and less susceptible to variation than evaluating % BCPs/total CD34+ cells. © 2017 International Clinical Cytometry Society.

Diagnosis of chronic lymphoproliferative disorders by flow cytometry using four-color combinations for immunophenotyping: A proposal of the brazilian group of flow cytometry (GBCFLUX).

BACKGROUND: Multiparametric flow cytometry (MFC) is a powerful tool for the diagnosis of hematological malignancies and has been useful for the classification of chronic lymphoproliferative disorders (CLPD) according to the WHO criteria. Following the purposes of the Brazilian Group of Flow Cytometry (GBCFLUX), the aim of this report was to standardize the minimum requirements to achieve an accurate diagnosis in CLPDs, considering the different economic possibilities of the laboratories in our country. Most laboratories in Brazil work with 4-fluorescence flow cytometers, which is why the GBCFLUX CLPD Committee has proposed 4-color monoclonal antibody (MoAb) panels. METHODS/RESULTS: Panels for screening and diagnosis in B, T and NK lymphoproliferative disorders were developed based on the normal differentiation pathways of these cells and the most frequent phenotypic aberrations. Important markers for prognosis and for minimal residual disease (MRD) evaluation were also included. The MoAb panels presented here were designed based on the diagnostic expertise of the participating laboratories and an extensive literature review. CONCLUSION: The 4-color panels presented to aid in the diagnosis of lymphoproliferative neoplasms by GBCFLUX aim to provide clinical laboratories with a systematic, step-wise, cost-effective, and reproducible approach to obtain an accurate immunophenotypic diagnosis of the most frequent of these disorders. © 2016 International Clinical Cytometry Society.

Evaluation of the Alere Pima™ for CD4+ T lymphocytes counts in HIV-positive outpatients in Southern Brazil.

CD4 + lymphocyte counts are routinely ordered during the early phases of antiretroviral therapy and for prophylaxis of opportunistic infections in HIV-positive patients. Flow cytometry is the standard methodology for CD4 counts in Brazilian reference laboratories. However, these laboratories are located in large cities, frequently distant from patients, thus limiting patient access and delaying results. We compared a point-of-care test with flow cytometry determination of CD4(+) T lymphocyte counts in HIV patients. We analysed 107 consecutive samples by both methods. Overall, the point-of-care test performed well, with excellent agreement between it and the standard method. Test results were concordant for patients with CD4(+) T lymphocyte values above and below 200 cells/mm (3). The performance characteristics obtained were sensitivity 94% (95% CI 89.5-98.5%), specificity 93% (95% CI 88.2-97.8%), positive predictive value 86% (95% CI 79.4-92.6%), and negative predictive value 97% (95% CI 94-100%). The high sensitivity and specificity of the point-of-care test methodology suggest its utility as an alternative method for rapid measurement of CD4(+) T lymphocytes in patients with limited access to reference laboratories, enabling prompt therapeutic intervention for patients at risk of progression to AIDS.

Biometric identification of capillariid eggs from archaeological sites in Patagonia.

Numerous eggs of capillariid nematodes have been found in coprolites from a wide range of hosts and in raptor pellets in archaeological samples from Patagonia. The structure and sculpture of the eggshell of these nematodes and their biometry are commonly used for identification. The aim of this study was to determine whether eggs of the genus Calodium with similar morphology, found in different archaeological samples from Patagonia, belong to the same species. For this purpose, capillariid eggs (N= 843) with thick walls and radial striations were studied by permutational multivariate analysis of variance (PERMANOVA). Eggs exhibiting similar shape and structure also showed similar biometry, regardless of the zoological origin of coprolites (P= 0.84), host diet (P= 0.19), character of the archaeological sites (P= 0.67) and chronology (P= 0.66). Thus, they were attributed to the same species. We suggest that an unidentified zoonotic species of the genus Calodium occurred in the digestive tract of a wide range of hosts in Patagonia during the Holocene and that both human and animal populations were exposed to this parasite during the Holocene in the study area.

PCR-RFLP genotyping of Toxoplasma gondii from chickens from Espírito Santo state, Southeast region, Brazil: new genotypes and a new SAG3 marker allele.

Brazil is one of the regions with the highest prevalences of Toxoplasma gondii in humans and animals. Because free-range chickens become infected by feeding from ground contaminated with oocysts, the prevalence of T. gondii in this host has been widely used as an indicator of the strains prevalent in the environment. The genetic variability among T. gondii isolates from different healthy and sick hosts all over the world has been recently studied. Three clonal genetic lineages (Types I, II and III) were initially recognised as predominant in Western Europe and the United States. T. gondii strains are genetically diverse in South America. In Brazil, recombination plays an important role in strain diversification. The objective of this study was to genetically characterise T. gondii isolates from free-range chickens from Espírito Santo state, Southeast region, Brazil, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). A total of 44 isolates among 47 previously described isolates (TgCkBr234-281) from free-range chickens were included in this study. Strain typing was performed using 12 PCR-RFLP markers: SAG1, SAG2, alt. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. Eleven genotypes were identified. Ten isolates (23%) were grouped into four novel genotypes. Four isolates, distributed in four counties, corresponded to the Type BrI lineage, the genotype found most frequently in Brazil. No clonal Types I, II or III lineages were found. Two novel genotypes were represented by single isolates. Unique alleles were identified for the markers SAG1, c22-8 and CS3, and for the first time, a unique allele was found for the marker SAG3. Although a large number of T. gondii genotypes have already been identified from a variety of animal hosts in Brazil, new genotypes are continuously identified from different animal species. This study confirmed the diversity of T. gondii in Brazil and demonstrates clonal Type I, II and III lineages are rare in this country.

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espírito Santo state, southeastern Brazil.

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espírito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo(®), Wama Diagnóstica, São Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of ≥ 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer ≥ 1:16) and 38.8% (198/510) by MAT (titer ≥ 1:25); concordance between IHAT and MAT was 81.6% (kappa index=0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64 (75%) seropositive (IHAT titers ≥ 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies.

Cochlear implant electrode array misplacement: a cautionary case report.

OBJECTIVE: To report a series of pitfalls and complications in a case of cochlear implantation. METHOD: Case report. RESULTS: An 11-year-old boy affected by auditory neuropathy underwent cochlear implantation. Intra-operative assessment was apparently consistent with correct insertion of the electrode array into the cochlea. However, subsequent high resolution computed tomography revealed that the entire electrode array was curled up within the vestibule. Revision surgery was complicated by cerebrospinal fluid leakage. A straight probe was repeatedly inserted into the internal auditory canal, before conversion to a canal wall down procedure and appropriate positioning of the electrode array. CONCLUSION: In this case, mild anteriorisation of the facial nerve created an awkward insertion angle for the electrode array via the retro-facial route, which may have triggered the described series of adverse events.

Polymorphisms of the promoter and exon 3 of the receptor for advanced glycation end products (RAGE) in Euro- and Afro-Brazilians.

The receptor for advanced glycation end products (RAGE or AGER), a member of the immunoglobulin superfamily, is involved in pathologies such as atherosclerosis and diabetes. Over 50 SNPs were reported for RAGE, among which were the promoter region polymorphisms -429T>C (rs1800625), -374T>A (rs1800624) and a 63-bp deletion (-407 to -345 bp), all related to increased RAGE expression. Additionally, in the exon 3, a putative site of binding ligands, the missense variation G82S (rs2070600) was associated with skin disorders in patients with diabetes. We have determined allele, genotype and haplotype frequencies of RAGE polymorphisms -429T>C, -374T>A, 63-bp deletion and G82S in Euro-Brazilians (n = 108) and Afro-Brazilians (n = 91), characterized according to the predominant ancestry of the individuals. The allele frequencies for Euro- and Afro-Brazilians were as follows: -429C, 12.5% vs. 12.1% (P = 0.90); -374A, 31.5% vs. 26.2% (P = 0.25); 63del, 0.0% vs. 3.8% (P = 0.004); and 82S, 1.9% vs. 0.6% (P = 0.24). Absolute linkage disequilibrium was found between the promoter polymorphisms -429T>C and -374T>A plus the 63-bp deletion (D'=1.000; P < 0.0001). The haplotype frequencies differed (P = 0.003) between Euro- and Afro-Brazilians. Our results showed that the frequencies of the 63-bp deletion were higher in Afro-Brazilians, while the other analysed polymorphisms were similarly distributed in the studied populations. The -374T>A plus 63-bp deletion polymorphism captures more than 80% of the haplotypic variation in the studied population.

Value of imaging and aspiration cytology in the diagnosis of oncocytic carcinoma.

Oncocytic carcinoma of the parotid gland is a rare neoplasm. To date 70 cases have been described in 55 reports. To the best of our knowledge the simultaneous occurrence of oncocytic carcinoma and second malignancy in another site (outside the parotid gland) has not been reported. An oncocytic carcinoma of the parotid gland is described in 56-year-old male with simultaneous breast cancer, emphasising the value of aspiration cytology and imaging procedures in the diagnosis of parotideal neoplasms.

First paleoparasitological results from late Holocene in Patagonian coprolites.

These are the first paleoparasitological results from the Late Holocene in Patagonian coprolites. Coprolites collected from layers dated at 3,480 and 2,740 yr before present (B.P.), from the archaeological site Cerro Casa de Piedra, were examined. The site is a hill located in a forest steppe ecotone in Perito Moreno National Park. The coprolites could belong to humans or to other carnivores such as canids. After rehydration and spontaneous sedimentation of the samples, the parasite contents were examined. Results obtained showed the common presence of eggs of a Calodium sp., eggs of other capillariids and trichostrongylids, oocysts of Eimeria macusaniensis (Apicomplexa), and eggs of taeniids (Cestoda). Although the generic identification of some parasites could not be provided, the presence of tapeworm eggs represents the first record for the Late Holocene in Patagonia and shows that parasitism by cestodes existed in the region in pre-Columbian times. Results indicate that in the Late Patagonic Holocene, zoonotic helminths may have been commonly present in the inhabitants of Patagonia. The parasites found in the coprolites allow us to deduce what these people were eating and, thus, indicate what other pathogens, such as viruses, bacteria, and parasitic protozoans, may have infected them via the same sources.

Vibrant Soundbridge for hearing restoration after chronic ear surgery.

INTRODUCTION: Middle ear surgery is primarily concerned with resolving the discharging pathology, in the case of chronic otitis media (COM), or with complete eradication, in case of cholesteatoma. Either of these procedures may require repeated surgeries, often resulting in severe mixed hearing impairment. A middle ear implant may be indicated in these cases instead of a hearing aid because the anatomical conditions in such cases often impede an adequate acoustic coupling. The objective of this study was to evaluate MED-EL Vibrant Soundbridge (VSB) implantation in patients with severe conductive and mixed hearing loss occurring after middle ear surgery for cholesteatoma or chronic otitis media (COM). MATERIALS AND METHODS: Over a 2-years period, the VSB system was implanted in 40 patients between 35 and 81 year old (mean: 59.5). Surgery was performed with comparable technique in 3 regional hospitals in Italy: Rovereto (n=16), Meran (n=12) and Tortona (n=12). The 40 candidates for implantation had a history of 1-5 previous surgeries. Of those, 20 patients suffered from COM and 20 from, cholesteatomas. The floating mass transducer (FMT) of the VSB was placed and stabilized on the round window niche in 32 cases; alternative positioning was necessary in 8 cases. Bone conduction (BC) was tested 1 day post-operatively. At 1 month post-surgery and between 6-9 months, open-field warble tones threshold in VSB-off and VSB-on conditions and open-field speech audiometry for words in quiet were conducted. RESULTS: Results of BC audiometry one day post-operatively showed no significant changes in hearing. Unaided mean pure tone average (PTA4) was 82.38 dB SPL with a mean speech recognition threshold (SRT) of 94.28 dB SPL. Results obtained after a minimum of three months post-operatively were evaluated in terms of aided thresholds and functional gain. At VSB activation, the mean PTA4 was 50.63 dB SPL with a mean SRT of 61.68 dB. After 6-9 months, the group had a mean PTA4 of 47.89 dB SPL and a mean SRT of 53.33 dB SPL. CONCLUSIONS: Implantation of the VSB with its direct driver of the inner ear fluids appears promising for auditory rehabilitation of severe mixed hearing loss associated with sequelae of cholesteatoma surgery. Patients' results improved over time, allowing us to assume a positive effect of consolidation of the coupling related to fibrosis. Results reported here refer to 6-9 months of observation and do not provide evidence of long term stability.

Microparticles in deep venous thrombosis, antiphospholipid syndrome and Factor V Leiden.

Microparticles (MPs) are blebs released from cellular surfaces during activation/apoptosis. They are procoagulant, pro-inflammatory and could contribute to pathogenesis of deep venous thrombosis (DVT). This study compared the number, cellular origin and procoagulant activity of MPs on DVT patients in different clinical situations: at diagnosis (n = 9, 5F/4M; mean age = 41.11), 1-3 years after warfarin withdrawal (n = 10, 7F/3M; mean age = 32.90), associated to antiphospholipid syndrome (APS; n = 11, 9F/2M; mean age = 33.82), or asymptomatic carriers of Factor V Leiden (FVL; n = 7, 7F/0M; mean age = 34.00) vs healthy controls (CTR). The quantification and characterization were performed by flow cytometry using CD235, CD61, CD45, CD31, CD14, CD45, anti-TF and Annexin V. The plasmatic procoagulant activity was investigated by prothrombin fragment 1 + 2 (F1 + 2) determination. The MPs procoagulant activity was analyzed by D-dimer (DD2) and Thrombin Generation Test (TGT) on a healthy pool of plasmas adjusted or not by their number (10,000 MPs). The MPs percentages were not different between the groups, but absolute number was increased in patients 1-3 years after warfarin withdrawal vs CTR (P = 0.02). There was no difference of the MPs cellular origin comparing patients to controls. TGT using 10,000 MPs was lower on these patients (P = 0.01). APS patients showed a reduction of plasmatic procoagulant activity (P = 0.004), but they were under warfarin therapy. DD2 in the presence of MPs, independently of its number, was higher in patients with DVT at diagnosis (P < 0.0001). MPs of patients with spontaneous DVT at diagnosis can promote coagulation activation demonstrated by increased DD2. Even the increased MPs from patients 1-3 years after thrombotic episode generated lower amount of thrombin, they can have a protective effect by activation of Protein C anticoagulant pathway.

Cadmium and zinc in Mar Chiquita coastal lagoon (Argentina): salinity effects on lethal toxicity in juveniles of the burrowing crab Chasmagnathus granulatus.

The large Argentine marine littoral zone is characterized by great number of wetlands and includes only one coastal lagoon, Mar Chiquita, which has been declared as a Biosphere Reserve by the Man and Biosphere Reserve Program from UNESCO. Its margins present populations of Chasmagnathus granulatus, a semiterrestrial crab distributed along wide salinity gradients that plays an important role as a key species within the corresponding trophic web. Dissolved cadmium (Cd) and zinc (Zn) concentrations present in this ecosystem were determined. Cadmium concentrations ranged between n.d. and 0.82 mug/L and zinc levels ranged between n.d. and 1224.38 mug/L within the mentioned coastal lagoon. Cd and Zn acute semistatic toxicity bioassays were carried out for 96 h on juvenile crabs of C. granulatus. LC(50) 96-h values were 2.24 mg Cd(2+)/L and 7.07 mg Zn(2+)/L at 5 psu, and 15.42 mg Cd(2+)/L and 11.41 mg Zn(2+)/L at 25 psu. Higher salinities resulted in lower metal toxicity. This effect was stronger for Cd than for Zn. C. granulatus juveniles LC(50) 96-h values determined for Cd were three to four orders of magnitude higher than the corresponding dissolved metal concentrations in the Mar Chiquita coastal lagoon; nevertheless, those Zn values determined were similar to several ones corresponding to natural water samples.

[Integration of the active middle ear implant Vibrant Soundbridge in total auricular reconstruction]. / Integration des aktiven Mittelohrimplantates in die plastische Ohrmuschelrekonstruktion.

BACKGROUND: Patients with high-grade microtia and atresia require a sophisticated and specific treatment. Apart from the plastic reconstruction of the auricle, in some cases hearing rehabilitation is medically indicated or is desired by the patients. The long-term results of simultaneous middle ear reconstruction with tympanoplasty are often inadequate owing to a persisting air-bone gap, and new techniques in hearing rehabilitation are needed for these patients. METHODS: We present three cases of unilateral atresia to illustrate a combined approach integrating hearing rehabilitation using the active middle ear implant Vibrant Soundbridge (VSB) into plastic auricular reconstruction. The VSB was attached to the stapes suprastructure via the titanium clip in two of these cases and in the third case a subfacial approach was used to attach it directly to the membrane of the round window. RESULTS: The air-bone gap was reduced to 17 dB, 14 dB and 0.25 dB HL. In free-field speech recognition tests at 65 dB SPL the patients achieved 100%, 90% and 100% recognition with the activated implant. No postoperative complications such as facial nerve paresis, vertigo or inner ear damage were found. CONCLUSIONS: The integration of active middle ear implants in auricular reconstruction opens up a new approach in complete hearing rehabilitation. The additional implantation of the VSB does not have any negative effect on the healing process or the cosmetic outcome of the auricular reconstruction.

Hyperpolarization-activated cyclic nucleotide-gated channel 1 is a molecular determinant of the cardiac pacemaker current I(f).

The pacemaker current I(f) of the sinoatrial node (SAN) is a major determinant of cardiac diastolic depolarization and plays a key role in controlling heart rate and its modulation by neurotransmitters. Substantial expression of two different mRNAs (HCN4, HCN1) of the family of pacemaker channels (HCN) is found in rabbit SAN, suggesting that the native channels may be formed by different isoforms. Here we report the cloning and heterologous expression of HCN1 from rabbit SAN and its specific localization in pacemaker myocytes. rbHCN1 is an 822-amino acid protein that, in human embryonic kidney 293 cells, displayed electrophysiological properties similar to those of I(f), suggesting that HCN1 can form a pacemaker channel. The presence of HCN1 in the SAN myocytes but not in nearby heart regions, and the electrophysiological properties of the channels formed by it, suggest that HCN1 plays a central and specific role in the formation of SAN pacemaker currents.

Cloning and expression pattern of a zebrafish homolog of forkhead activin signal transducer (FAST), a transcription factor mediating Nodal-related signals.

Forkhead activin signal transducer (FAST) is a member of the winged-helix family of DNA-binding proteins that has been implicated in mesoderm induction and left-right axis specification during embryonic development in Xenopus and mouse. We have cloned and characterized a zebrafish FAST homolog. Zebrafish fast is expressed maternally and zygotically. Transcripts start regionalizing and decline in level during gastrulation. During somitogenesis, fast is expressed bilaterally in the lateral plate mesoderm, like its mouse homolog. In addition, zebrafish fast is also expressed bilaterally in the dorsal diencephalon, where the nodal-related cyclops gene is only expressed on the left side. It remains to be demonstrated whether FAST expression in the brain can mediate Nodal-induced asymmetric development.