Hair follicle-resident progenitor cells are a major cellular contributor to heterotopic subcutaneous ossifications in a mouse model of Albright hereditary osteodystrophy.

Heterotopic ossifications (HOs) are the pathologic process by which bone inappropriately forms outside of the skeletal system. Despite HOs being a persistent clinical problem in the general population, there are no definitive strategies for their prevention and treatment due to a limited understanding of the cellular and molecular mechanisms contributing to lesion development. One disease in which the development of heterotopic subcutaneous ossifications (SCOs) leads to morbidity is Albright hereditary osteodystrophy (AHO). AHO is caused by heterozygous inactivation of GNAS, the gene that encodes the α-stimulatory subunit (Gαs) of G proteins. Previously, we had shown using our laboratory's AHO mouse model that SCOs develop around hair follicles (HFs). Here we show that SCO formation occurs due to inappropriate expansion and differentiation of HF-resident stem cells into osteoblasts. We also show in AHO patients and mice that Secreted Frizzled Related Protein 2 (SFRP2) expression is upregulated in regions of SCO formation and that elimination of Sfrp2 in male AHO mice exacerbates SCO development. These studies provide key insights into the cellular and molecular mechanisms contributing to SCO development and have implications for potential therapeutic modalities not only for AHO patients but also for patients suffering from HOs with other etiologies.

Endothelial to mesenchymal Notch signaling regulates skeletal repair.

We present a transcriptomic analysis that provides a better understanding of regulatory mechanisms within the healthy and injured periosteum. The focus of this work is on characterizing early events controlling bone healing during formation of periosteal callus on day 3 after fracture. Building on our previous findings showing that induced Notch1 signaling in osteoprogenitors leads to better healing, we compared samples in which the Notch 1 intracellular domain is overexpressed by periosteal stem/progenitor cells, with control intact and fractured periosteum. Molecular mechanisms and changes in skeletal stem/progenitor cells (SSPCs) and other cell populations within the callus, including hematopoietic lineages, were determined. Notably, Notch ligands were differentially expressed in endothelial and mesenchymal populations, with Dll4 restricted to endothelial cells, whereas Jag1 was expressed by mesenchymal populations. Targeted deletion of Dll4 in endothelial cells using Cdh5CreER resulted in negative effects on early fracture healing, while deletion in SSPCs using α-smooth muscle actin-CreER did not impact bone healing. Translating these observations into a clinically relevant model of bone healing revealed the beneficial effects of delivering Notch ligands alongside the osteogenic inducer, BMP2. These findings provide insights into the regulatory mechanisms within the healthy and injured periosteum, paving the way for novel translational approaches to bone healing.

Hematopoietic and stromal DMP1-Cre labeled cells form a unique niche in the bone marrow.

Skeletogenesis and hematopoiesis are interdependent. Niches form between cells of both lineages where microenvironmental cues support specific lineage commitment. Because of the complex topography of bone marrow (BM), the identity and function of cells within specialized niches has not been fully elucidated. Dentin Matrix Protein 1 (DMP1)-Cre mice have been utilized in bone studies as mature osteoblasts and osteocytes express DMP1. DMP1 has been identified in CXCL12+ cells and an undefined CD45+ population. We crossed DMP1-Cre with Ai9 reporter mice and analyzed the tdTomato+ (tdT+) population in BM and secondary hematopoietic organs. CD45+tdT+ express myeloid markers including CD11b and are established early in ontogeny. CD45+tdT+ cells phagocytose, respond to LPS and are radioresistant. Depletion of macrophages caused a significant decrease in tdT+CD11b+ myeloid populations. A subset of CD45+tdT+ cells may be erythroid island macrophages (EIM) which are depleted after G-CSF treatment. tdT+CXCL12+ cells are in direct contact with F4/80 macrophages, express RANKL and form a niche with B220+ B cells. A population of resident cells within the thymus are tdT+ and express myeloid markers and RANKL. In conclusion, in addition to targeting osteoblast/osteocytes, DMP1-Cre labels unique cell populations of macrophage and stromal cells within BM and thymus niches and expresses key microenvironmental factors.

Discrimination and Perceived Cultural Mismatch Increase Status-Based Identity Uncertainty.

Periods of social mobility, such as attending college, can challenge one's status-based identity, leading to uncertainty around one's status in society. Status uncertainty is associated with poorer well-being and academic outcomes. Little is known, however, about what experiences lead to status uncertainty. The current longitudinal study investigated discrimination experiences and cultural mismatch as predictors of status uncertainty. We propose that discrimination indirectly predicts increased status uncertainty by increasing perceived cultural mismatch with the university. Participants were Latinx college students, all of whom were low-income and/or first generation to college. Discrimination experiences were measured at the end of participants' first year. Cultural mismatch and status uncertainty were measured at the end of Year 2. Status uncertainty was measured again at the end of Year 3. Results indicated that students who experienced more frequent discrimination felt more cultural mismatch 1 year later, and, in turn, reported increased status uncertainty over the following year.

Lineage Tracing of RGS5-CreER-Labeled Cells in Long Bones During Homeostasis and Injury.

Regulator of G protein signaling 5 (RGS5) is a GTPase activator for heterotrimeric G-protein α-subunits, shown to be a marker of pericytes. Bone marrow stromal cell population (BMSCs) is heterogeneous. Populations of mesenchymal progenitors, cells supportive of hematopoiesis, and stromal cells regulating bone remodeling have been recently identified. Periosteal and bone marrow mesenchymal stem cells (MSCs) are participating in fracture healing, but it is difficult to distinguish the source of cells within the callus. Considering that perivascular cells exert osteoprogenitor potential, we generated an RGS5 transgenic mouse model (Rgs5-CreER) which when crossed with Ai9 reporter animals (Rgs5/Tomato), is suitable for lineage tracing during growth and post-injury. Flow cytometry analysis and histology confirmed the presence of Rgs5/Tomato+ cells within CD31+ endothelial, CD45+ hematopoietic, and CD31-CD45- mesenchymal/perivascular cells. A tamoxifen chase showed expansion of Rgs5/Tomato+ cells expressing osterix within the trabeculae positioned between mineralized matrix and vasculature. Long-term chase showed proportion of Rgs5/Tomato+ cells contributes to mature osteoblasts expressing osteocalcin. Following femoral fracture, Rgs5/Tomato+ cells are observed around newly formed bone within the BM cavity and expressed osterix and osteocalcin, while contribution within periosteum was low and limited to fibroblastic callus with very few positive chondrocytes. In addition, BM injury model confirmed that RGS5-Cre labels population of BMSCs expands during injury and participates in osteogenesis. Under homeostatic conditions, lineage-traced RGS5 cells within the trabecular area demonstrate osteoprogenitor capacity that in an injury model contributes to new bone formation primarily within the BM niche.

Inhibition of CGRP signaling impairs fracture healing in mice.

Calcitonin gene-related peptide (CGRP) is a neuropeptide produced by sensory nerves and functions as a pain sensor. It acts by binding to the calcitonin-like receptor (CLR, protein; Calcrl, gene). CGRP inhibition has been recently introduced as therapeutic treatment of migraine-associated pain. Previous studies have shown that CGRP stimulates bone formation. The aim of our study is to determine whether the inhibition of CGRP signaling negatively impacted fracture healing. Using α-smooth muscle actin (αSMA) Cre animals crossed with Ai9 reporter mice, we showed that CGRP-expressing nerves are near αSMA + cells in the periosteum. In vitro experiments revealed that periosteal cells express Calcrl and receptor activity modifying protein 1; and CGRP stimulation increased periosteal cell proliferation. Using a tamoxifen-inducible model αSMACre/CLRfl/fl , we targeted the deletion of CLR to periosteal progenitor cells and examined fracture healing. Microcomputed tomography of fractured femurs showed a reduction in bone mass in αSMACre+/CLRfl/fl female mice relative to controls and callus volume in males. Pharmacological CGRP-CLR inhibition was achieved by subcutaneous delivery of customized pellets with small molecule inhibitor olcegepant (BIBN-4096) at a dose of 10 µg/day. BIBN-4096-treated C57BL/6J mice had a higher latency toward thermal nociception than placebo-treated mice, indicating impaired sensory function through CGRP inhibition. CGRP inhibition also resulted in reduced callus volume, bone mass, and bone strength compared to placebo controls. These results indicate that inhibiting CGRP by deleting CLR or by using BIBN-4096, contributes to delayed bone healing.

Novel population of human monocyte and osteoclast progenitors from pluripotent stem cells and peripheral blood.

Osteoclasts are multinuclear cells of monocytic lineage, with the ability to resorb bone. Studies in mouse have identified bone marrow clonal progenitors able to generate mature osteoclast cells (OCs) in vitro and in vivo. These osteoclast progenitors (OCPs) can also generate macrophages and dendritic cells. Interestingly, cells with equivalent potential can be detected in periphery. In humans, cells with OCP activity have been identified in bone marrow and periphery; however, their characterization has not been as extensive. We have developed reproducible methods to derive, from human pluripotent stem cells, a population containing monocyte progenitors able to generate functional OCs. Within this population, we have identified cells with monocyte and osteoclast progenitor activity based on CD11b and CD14 expression. A population double positive for CD11b and CD14 contains cells with expected osteoclastic potential. However, the double negative (DN) population, containing most of the hematopoietic progenitor activity, also presents a very high osteoclastic potential. These progenitor cells can also be differentiated to macrophage and dendritic cells. Further dissection within the DN population identified cells bearing the phenotype CD15-CD115+ as the population with highest monocytic progenitor and osteoclastic potential. When similar methodology was used to identify OCPs from human peripheral blood, we confirmed a published OCP population with the phenotype CD11b+CD14+. In addition, we identified a second population (CD14-CD11bloCD115+) with high monocytic progenitor activity that was also able to form osteoclast like cells, similar to the 2 populations identified from pluripotent stem cells.

First Report of Alternaria japonica, a causal agent of black spot, on kale in South Carolina, United States.

In January 2020, charcoal gray, dull lesions were observed on leaves of organic kale (Brassica oleracea var. acephala) cv. Darkibor in two fields in Lexington County, South Carolina, the county with the most acres of leafy brassicas in the state. Leaf spots, also visible on the leaf underside, covered <5% of the leaf area. No spores were present. Portions of leaf spots from eight leaves, four per field, were cultured on one-quarter-strength potato dextrose agar (PDA/4). Eleven isolates of Alternaria spp. were recovered. Isolates ALT12 and UL3 were cultured in A. solani medium and DNA was extracted (Maiero et al. 1991). The internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (tef1), RNA polymerase second largest subunit (rpb2), and Alternaria major allergen (Alt a 1) genes were amplified with the primer pairs V9G/ITS4, EF1-728F/EF1-986R, RPB2-5F2/FRPB2-7cR, and Alt-for/Alt-rev, respectively, and sequenced (Woudenberg et al. 2014). Sequences for isolates ALT12 and UL3, collected from different leaves in the same field, were identical to each other and to isolate AC97 (ITS accession number: LC440597; tef1: LC482018; rpb2: LC476803; Alt a 1: LC481628) of A. japonica Yoshii (Nishikawa and Nakashima 2020). ITS, tef1, repb2, and Alta a 1 sequences for each isolate were deposited in GenBank under the accessions MW374952, MW389653, MW389655, and MW389657 for ALT12 and MW374951, MW389652, MW389654, and MW389656 for UL3, respectively. Conidia of A. japonica (20 of ALT12, 10 of UL3) produced by 7-day-old cultures on Spezieller Nährstoffarmer Agar measured 62.1 ± 11.4 x 18.8 ± 2.2 µm (standard deviation). Median numbers of transverse and longitudinal septae were 6 (4 to 8) and 2 (1 to 3), respectively. Conidia formed singly or in chains of two. Cells were constricted around the transverse septae (Nishikawa and Nakashima 2020; Woudenburg et al. 2014). Chlamydospores were present in cultures of ALT12. ALT12 was pathogenic on kale cv. Darkibor and Winterbor inoculated in a greenhouse following procedures of Al-Lami et al. (2019). Four replicate pots with two plants each were used; plants were 6, 9, and 5 weeks old in trials 1, 2, and 3, respectively. The oldest three leaves of each plant were spray inoculated with a suspension of 5 x 105 conidia/ml; noninoculated control plants were sprayed with water. All plants were kept for 48 h at 100% RH, then moved to a bench in a greenhouse held at 21/16°C day/night temperatures. The second and third oldest leaves were rated 13 days after inoculation. Small gray or black spots developed on inoculated leaves and petioles in all trials, and on one noninoculated leaf in trial one. Disease incidence on inoculated leaves (73.1%) was greater than on noninoculated leaves (0.05%) (P<0.0001). Cultivars did not differ in susceptibility (P=0.12). Portions of lesions on inoculated leaves and portions of noninoculated leaves were cultured onto PDA/4 amended with antibiotics (Keinath 2013). A. japonica was reisolated from 46 of 50 inoculated leaf blades; 22 of 28 inoculated petioles; and 1 of 8, 0 of 8, and 0 of 7 noninoculated leaves in the three trials, respectively. Growers in South Carolina consider black spot, or Alternaria leaf spot, the most important fungal disease on organic kale. The presence of a second causal agent in addition to A. brassicae may increase disease occurrence. A. japonica previously was reported on arugula in California (Tidwell et al. 2014). This is the first report of A. japonica in the eastern United States.

Perivascular osteoprogenitors are associated with transcortical channels of long bones.

Bone remodeling and regeneration are dependent on resident stem/progenitor cells with the ability to replenish mature osteoblasts and repair the skeleton. Using lineage tracing approaches, we identified a population of Dmp1+ cells that reside within cortical bone and are distinct from osteocytes. Our aims were to characterize this stromal population of transcortical perivascular cells (TPCs) in their resident niche and evaluate their osteogenic potential. To distinguish this population from osteoblasts/osteocytes, we crossed mice containing inducible DMP1CreERT2/Ai9 Tomato reporter (iDMP/T) with Col2.3GFP reporter (ColGFP), a marker of osteoblasts and osteocytes. We observed iDMP/T+;ColGFP- TPCs within cortical bone following tamoxifen injection. These cells were perivascular and located within transcortical channels. Ex vivo bone outgrowth cultures showed TPCs migrated out of the channels onto the plate and expressed stem cell markers such as Sca1, platelet derived growth factor receptor beta (PDGFRß), and leptin receptor. In a cortical bone transplantation model, TPCs migrate from their vascular niche within cortical bone and contribute to new osteoblast formation and bone tube closure. Treatment with intermittent parathyroid hormone increased TPC number and differentiation. TPCs were unable to differentiate into adipocytes in the presence of rosiglitazone in vitro or in vivo. Altogether, we have identified and characterized a novel stromal lineage-restricted osteoprogenitor that is associated with transcortical vessels of long bones. Functionally, we have demonstrated that this population can migrate out of cortical bone channels, expand, and differentiate into osteoblasts, therefore serving as a source of progenitors contributing to new bone formation.

Slip-Based Coding of Local Shape and Texture in Mouse S1.

Tactile objects have both local geometry (shape) and broader macroscopic texture, but how these different spatial scales are simultaneously encoded during active touch is unknown. In the whisker system, we tested for a shared code based on localized whisker micromotions (stick-slips) and slip-evoked spikes. We trained mice to discriminate smooth from rough surfaces, including ridged gratings and sandpaper. Whisker slips locked to ridges and evoked temporally precise spikes (<10 ms jitter) in somatosensory cortex (S1) that could resolve ridges with ∼1 mm accuracy. Slip-sensitive neurons also encoded touch and texture. On rough surfaces, both slip-evoked spikes and an additional non-slip signal elevated mean firing rate, allowing accurate rough-smooth texture decoding from population firing rate. Eighteen percent of neurons were selective among rough surfaces. Thus, slips elicit spatially and temporally precise spiking in S1 that simultaneously encodes local shape (ridges) and is integrated into a macroscopic firing rate code for roughness.

Adaptive Control in Passive rehabilitation routines using ELLTIO / Control Adaptable en rutinas de rehabilitación pasiva utilizando ELLTIO

ABSTRACT: The Exoskeleton for Lower Limb Training with Instrumented Orthosis (ELLTIO) is a mechatronic device that can be used to assist in passive kinesitherapy to increase human muscles strength and resistance [1]. This paper presents an alternative for passive rehabilitation process using an exoskeleton for knee and ankle. The main idea is assist a pro fessional physiotherapist in the design and performance of exercises routines for his patients using the prototype. The knee and ankle joint's movements are recorded and storage during the exercises to propose a similar computer generated trajectories which the exoskeleton on should follow. An adaptive controller is implemented to track the trajectories and adapt the user parameters.

RESUMEN: El exoesqueleto para el entrenamiento de miembros inferiores con órtesis instrumentada (ELLTIO) por sus siglas en ingles "Exoskeleton for Lower Limb Training with Instrumented Orthosis" es un dispositivo mecatrónico que se puede utilizar para ayudar en la fisioterapia pasiva para aumentar la fuerza y resistencia de los músculos humanos. En este trabajo se presenta una alternativa para el proceso de rehabilitación pasiva utilizando un exoesqueleto de rodilla y tobillo. La idea principal es ayudar a un fisioterapeuta profesional en el diseño y ejecución de rutinas de ejercicios para sus pacientes utilizando el prototipo. Los movimientos de la articulación de la rodilla y el tobillo se registran y se almacenan durante los ejercicios para proponer trayectorias similares generadas por computadora que el exoesqueleto debe seguir. Se implementa un controlador adaptativo para rastrear las trayectorias y adaptar los parámetros del usuario.

Carbon dioxide laser ablation of basal cell carcinoma with visual guidance by reflectance confocal microscopy: a proof-of-principle pilot study.

BACKGROUND: Laser ablation is an alternative, nonsurgical treatment modality for low-risk basal cell carcinoma (BCC). However, lack of confirmative tumour destruction or residual tumour presence has been a limiting factor to its adoption. Reflectance confocal microscopy (RCM) provides noninvasive, cellular-level resolution imaging of the skin and is capable of identifying tumour. OBJECTIVES: To evaluate the use of RCM to guide carbon dioxide (CO2 ) laser ablation of BCC, confirm destruction and correlate findings with histology. METHODS: RCM was used preablation to evaluate for features of BCC. Ablation was performed with a CO2 laser, and the response rapidly assessed using handheld RCM to evaluate for residual tumour. Confirmative pathology was used to verify confocal imaging. RESULTS: Preablation RCM imaging identified tumour with features not identified on normal, surrounding skin. Postablation, RCM documented complete removal of tumour in six cases and residual tumour in two. Histological examination identified the ablated area and confirmed clearance of tumour in the six aforementioned cases and corroborated confocal findings for residual tumour in the other two cases. CONCLUSIONS: We report successful treatment of superficial and nodular BCC using CO2 laser ablation augmented by RCM imaging for preablation guidance and verification of tumour removal postablation. Akin to complete circumferential and deep margin control techniques, using RCM helps to map peripheral and deep BCC margins to hone in on areas exhibiting persistent tumour after ablation. CO2 laser ablation visually guided by RCM can help circumvent previously cited limiting factors of laser ablation for tumour destruction by providing cellular-level resolution imaging of tumour and margin assessment in between each laser pass and postablation.

Transcriptional regulation of IL-15 expression during hematopoiesis.

Dendritic cells (DCs) are the most commonly studied source of the cytokine IL-15. Using an IL-15 reporter transgenic mouse, we have recently shown previously unappreciated differences in the levels of IL-15 expressed by subsets of conventional DCs (CD8⁺ and CD8⁻). In this study, we show that IL-15 promoter activity was differentially regulated in subsets of hematopoietically derived cells with IL-15 expression largely limited to myeloid lineages. In contrast, mature cells of the lymphoid lineages expressed little to no IL-15 activity. Surprisingly, we discovered that hematopoietic stem cells (lineage⁻Sca-1⁺c-Kit⁺) expressed high levels of IL-15, suggesting that IL-15 expression was extinguished during lymphoid development. In the case of T cells, this downregulation was Notch-dependent and occurred in a stepwise pattern coincident with increasing maturation and commitment to a T cell fate. Finally, we further demonstrate that IL-15 expression was also controlled throughout DC development, with key regulatory activity of IL-15 production occurring at the pre-DC branch point, leading to the generation of both IL-15⁺CD8⁺ and IL-15(⁻/low)CD8⁻ DC subsets. Thus, IL-15 expression is coordinated with cellular fate in myeloid versus lymphoid immune cells.

A novel population of cells expressing both hematopoietic and mesenchymal markers is present in the normal adult bone marrow and is augmented in a murine model of marrow fibrosis.

Bone marrow (BM) fibrosis is a feature of severe hyperparathyroidism. Consistent with this observation, mice expressing constitutively active parathyroid hormone (PTH)/PTH-related peptide receptors (PPR) in osteoblasts (PPR*Tg) display BM fibrosis. To obtain insight into the nature of BM fibrosis in such a model, a double-mutant mouse expressing constitutively active PPR and green fluorescent protein (GFP) under the control of the type I collagen promoter (PPR*Tg/GFP) was generated. Confocal microscopy and flow cytometry revealed the presence of a cell population expressing GFP (GFP(+)) that was also positive for the hematopoietic marker CD45 in the BM of both PPR*Tg/GFP and control animals. This cell population was expanded in PPR*Tg/GFP. The existence of cells expressing both type I collagen and CD45 in the adult BM was confirmed by IHC and fluorescence-activated cell sorting. An analysis of total RNA extracted from sorted GFP(+)CD45(+) cells showed that these cells produced type I collagen and PTH/PTH-related peptide receptor and receptor activator for NF-κB mRNAs, further supporting their features of being both mesenchymal and hematopoietic lineages. Similar cells, known as fibrocytes, are also present in pathological fibroses. Our findings, thus, indicate that the BM is a permissive microenvironment for the differentiation of fibrocyte-like cells and raise the possibility that these cells could contribute to the pathogenesis of BM fibrosis.

cPLA2 is protective against COX inhibitor-induced intestinal damage.

Cytosolic phospholipase A(2) (cPLA(2)) is the rate-limiting enzyme responsible for the generation of prostaglandins (PGs), which are bioactive lipids that play critical roles in maintaining gastrointestinal (GI) homeostasis. There has been a long-standing association between administration of cyclooxygenase (COX) inhibitors and GI toxicity. GI injury is thought to be induced by suppressed production of GI-protective PGs as well as direct injury to enterocytes. The present study sought to determine how pan-suppression of PG production via a genetic deletion of cPLA(2) impacts the susceptibility to COX inhibitor-induced GI injury. A panel of COX inhibitors including celecoxib, rofecoxib, sulindac, and aspirin were administered via diet to cPLA(2)(-/-) and cPLA(2)(+/+) littermates. Administration of celecoxib, rofecoxib, and sulindac, but not aspirin, resulted in acute lethality (within 2 weeks) in cPLA(2)(-/-) mice, but not in wild-type littermates. Histomorphological analysis revealed severe GI damage following celecoxib exposure associated with acute bacteremia and sepsis. Intestinal PG levels were reduced equivalently in both genotypes following celecoxib exposure, indicating that PG production was not likely responsible for the differential sensitivity. Gene expression profiling in the small intestines of mice identified drug-related changes among a panel of genes including those involved in mitochondrial function in cPLA(2)(-/-) mice. Further analysis of enterocytic mitochondria showed abnormal morphology as well as impaired ATP production in the intestines from celecoxib-exposed cPLA(2)(-/-) mice. Our data demonstrate that cPLA(2) appears to be an important component in conferring protection against COX inhibitor-induced enteropathy, which may be mediated through affects on enterocytic mitochondria.

Fast high performance liquid chromatography and ultraviolet-visible quantification of principal phenolic antioxidants in fresh rosemary.

An improved HPLC method is reported for the determination of rosemary's principal phenolic antioxidants, rosmarinic and carnosic acids, providing a fast and simultaneous determination for both of them by using a solid phase column. The analysis was performed with fresh methanolic extractions of Rosmarinus officinalis. To quantify the amount of antioxidants in a fast and reproducible way by means of UV-vis absorption measurements, a spectrophotometric multi-wavelength calibration curve was constructed based on the antioxidant contents obtained with the recently developed HPLC method. This UV-vis methodology can be extended to the determination of other compounds and herbs if the restrictions mentioned in the text are respected.

Cerebral tumor-like American trypanosomiasis in acquired immunodeficiency syndrome.

Cerebral tumor-like American trypanosomiasis (CTLAT) is an uncommon complication of Chagas' disease, observed only in immunosuppressed patients. We assessed 10 human immunodeficiency virus-positive patients with Chagas' disease who presented with CTLAT. All patients had neurological involvement and 6 developed intracranial hypertension. Neuroimaging studies showed supratentorial lesions in 9 patients, being single in 8. One case had infratentorial and supratentorial lesions. Low CD4+ cell counts were observed in all the cases and in 6 of them CTLAT was the first manifestation of acquired immunodeficiency syndrome. Serological tests for Chagas' disease were positive in 6 of 8 patients. Trypanosoma cruzi was identified in all brain specimens and in three cerebrospinal fluid samples. CTLAT should be considered in the differential diagnosis of intracranial mass lesions in human immunodeficiency virus-positive patients and should be added to the list of acquired immunodeficiency syndrome-defining illnesses.

La prueba dot-ELISA para la detección de campo de la leishmaniasis visceral en Honduras / Field use of the dot-ELISA test for visceral leishmaniasis in Honduras

En una zona endémica de la República de Honduras, se llevó a cabo un ensayo sobre el terreno de la prueba de inmunoabsorbencia ligada a la enzima efectuada en disco de nitrocelulosa (dot-ELISA), como microtécnica rápida y susceptible de interpretarse a simple vista, para el diagnóstico serológico de la leishmaniasis visceral en el hombre. De los 305 sujetos investigados usando una dilución del suero de 1:32, se observaron reacciones positivas en ocho de los nueve casos de leishmaniasis visceral diagnosticada mediante estudio parasitológico que habían recibido tratamiento, en 13 de los 45 familiares de pacientes (grupo expuesto a un gran riesgo) y en ocho de los 244 niños seleccionados al azar en la zona endémica. Se observaron reacciones cruzadas en uno de los tres niños con leishmaniasis cutánea confirmada por estudio parasitológico y en tres de los cuatro adultos con resultados serológicos positivos para enfermedad de Chagas. La determinación de los títulos de punto final de los sueros de los pacientes con leishmaniasis visceral produjo títulos recíprocos que fluctuaron entre 512 y 8 192, inferiores a los que por lo general se encuentran en casos activos no tratados. Esta prueba no requiere instalaciones eléctricas y todos los materiales pueden transportarse fácilmente a pie. Es un procedimiento rápido, sencillo, poco costoso y, no obstante, sensible y relativamente específico en las condiciones propias del terreno. Podría resultar un instrumento valioso para los servicios de atención primaria de salud y para las encuestas epidemiológicas en las numerosas zonas endémicas donde actualmente no es posible efectuar pruebas serológicas

Development of a compressed product made with sardine.

The per capita consumption of marine products is very low in Mexico, averaging less than 4 g/day. This fact has been partially attributed to the costly techniques used in their preservation, which result in high market prices unaffordable for large segments of the population. Previous research lead to the development of pressed and salted patties based on lean fish species, the low cost and easy preservation of which would contribute to a higher fish consumption among the low socio-economic strata of the Mexican population. The present work was directed to adapt the procedure to sardine, which is more abundant and less expensive than lean fish species. Since defatting the sardine lead to poor sensorial characteristics of the patties, measures were taken to protect the fat from oxidation, through the use of BHT and citric acid. The best results were obtained with descaled sardine, and with the addition of 8% NaCl, 10% corn flour and a condiment mixture. The resulting product had 32% of high-quality protein and a shelf life of at least six months under environmental conditions. Its cost per gram of protein was one-third lower than the price of fresh or canned sardine. Sensorial tests revealed an acceptability of 82%.