A Diagnostic Algorithm for Detection of Leishmania spp. in Human Fresh and Fixed Tissue Samples.

Leishmaniasis is an important travel-related parasitic infection in the United States. Treatment regimens vary by Leishmania species and require an accurate diagnosis. The sensitivity and specificity of diagnostic methods depend on the type and condition of specimen analyzed. To identify the best algorithm for detection of parasites in fresh and fixed tissue samples, we evaluated parasite cultures, two PCR methods, and Leishmania immunohistochemistry (IHC) in samples received by the CDC from 2012 through 2019. The sensitivity and specificity of IHC assays were evaluated in fresh specimens tested. Diagnostic accuracy for formalin-fixed tissue was evaluated by using PCR-based methods and IHC. Of 100 suspected cases with fresh tissue available, Leishmania spp. infection was identified by PCR in 56% (56/100) of specimens; from these, 80% (45/56) were positive by parasite culture and 59% (33/56) by IHC. Of 420 possible cases where only fixed specimens were available, 58% (244/420) were positive by IHC and/or PCR. Of these, 96% (235/420) were positive by IHC and 84% (204/420) by PCR-based methods. Overall parasite detection using all methodologies was similar for fresh and formalin-fixed tissue specimens (56% versus 58%, respectively). Although PCR-based methods were superior for diagnosis of leishmaniasis and species identification in fresh samples, IHC in combination with PCR increased the accuracy for Leishmania spp. detection in fixed samples. In conclusion, PCR is the most effective method for detecting Leishmania infection in fresh tissue samples, whereas for formalin-fixed samples, IHC and PCR-based methods should be used in combination.

Daily variability of pH and temperature in seawater from a near-pristine oceanic atoll, Southwest Atlantic.

The study of pH and temperature variability in reef environments, and the underlying processes that control this variability, is of great importance for ocean acidification research. Therefore, in the reef environment of Rocas Atoll, we conducted continuous monitoring of pH and temperature and periodic sampling of carbonate chemistry, and we hypothesize that seawater temperature is not the determining factor in the daily variability of pH at this atoll. Our results showed that the seawater of the atoll presented a high daily variability in pH, [H+], and temperature. The cycles of variations occurred primarily with a periodicity of ∼24 h, related to the daily light cycle, and secondarily with a periodicity of ∼12 h, associated with the semi-diurnal tidal cycles of the atoll. The results indicate that the relative balance of net organic carbon metabolism is the main process modulating carbonate chemistry on the atoll throughout the day.

Heinrich Ewald Hering's discovery of the heart pacemaker: Hering, Tawara and Aschoff's search for its morphological basis, the sinoatrial node, and why they failed.

Two groups of investigators investigated the heart pacemaker and its morphological basis in the early twentieth century. The first group was formed by Henrich Ewald Hering (physiologist), Sunao Tawara and Ludwig Aschoff (morphologists). The second group was composed of James Mackenzie (general practitioner and clinical investigator), Arthur Keith and Martin Flack (morphologists). These groups were formed almost at the same time in 1903. Their work resulted in the discovery of the atrioventricular node and Purkinje network (Sunao Tawara, in 1906), heart pacemaker (H E Hering, in 1907) and sinoatrial node (Keith and Flack, in 1907). Here, it is shown how the interconnections of the concurrent works of these groups resulted in the discovery not only of the function, but also of the structure of the sinoatrial node.

Evidence for concealed fasciculo-ventricular connections as revealed by His bundle pacing.

BACKGROUND: It is almost 100 years ago since Mahaim described the so-called paraspecific connections between the ventricular conduction axis and the crest of the muscular ventricular septum, believing such pathways to be ubiquitous. These pathways, however, have yet to be considered as potential pathways for septal activation during His bundle pacing. MATERIALS: So as to explore the hypothesis that specialised septal pathways might provide the substrate for septal activation during His bundle pacing, we compared the findings from 22 serially sectioned histological datasets and 34 different individuals undergoing His bundle pacing. RESULTS: We found histologically specialised pathways connecting the branching component of the atrioventricular conduction axis with the crest of the muscular ventricular septum in almost four-fifths of the histological datasets. In 32 of 34 patients undergoing His bundle pacing, the QRS complex closely resembled published images of known conduction through fasciculo-ventricular pathways. In only two patients was a delta wave not seen at any pacing voltages. Capture of these connections varied according to pacing voltage, a finding which correlated with the distance of the pathways from the site of penetration of the ventricular conduction axis. Ventricular activation times remained normal in the presence of the delta wave at higher pacing voltage but were prolonged at lower voltages. CONCLUSIONS: Our histologic findings confirm fasciculo-ventricular connections, initially described by Mahaim as being paraspecific, are likely ubiquitous. Analysis of 12-lead electrocardiograms leads us to conclude that fasciculo-ventricular pathways, concealed during sinus rhythm, become manifest with His bundle pacing.

Miniseries 1-Part II: the comparative anatomy of the atrioventricular conduction axis.

AIMS: The arrangement of the conduction axis is markedly different in various mammalian species. Knowledge of such variation may serve to question the validity of using animals as prospective models for design of systems for clinical use. METHODS AND RESULTS: We compared the arrangement of the atrioventricular conduction axis in human, murine, canine, porcine, and bovine hearts, examining serially sectioned datasets from 20 human, 16 murine, 3 porcine, 5 canine, and 1 bovine hearts. We also analysed computed tomographic datasets obtained from bovines and one human heart. Unlike the situation in the human heart, there is no formation of an atrioventricular fibrous membranous septum in the murine, canine, porcine, nor bovine hearts. Canine, porcine, and bovine hearts also lack an infero-septal recess, when defined as a fibrous plate supporting the buttress of the atrial septum. In these species, half of the non-coronary leaflet is directly opposed to the ventricular septal surface. CONCLUSION: There is a long right-sided non-branching component of the axis, which skirts the attachment of the non-coronary sinus of the aortic root. In the bovine heart, moreover, the left bundle branch usually extends intramyocardially as a solitary tape before surfacing and ramifying on the left ventricular septal surface. The difference in the atrioventricular conduction axis between species may influence the anatomical substrates for atrioventricular re-entry tachycardia, as well as providing inferences for assessing the risks of transcatheter implantation of the aortic valve. Further studies are now needed to assess these possibilities.

The atrioventricular conduction axis and the aortic root-Inferences for transcatheter replacement of the aortic valve.

Conduction problems still occur following transcatheter aortic valvar replacement. With this in mind, we have assessed the relationship of the conduction axis to the aortic root. We used serial histological sections, made perpendicular to the base of the triangle of Koch in nine hearts, and perpendicular to the aortic root in 11 hearts. We first defined the extent of the fibrous tissues forming the boundaries of an infero-septal recess of the subaortic outflow tract, found in all datasets but one. When the recess was present, the axis penetrated through its rightward wall, giving rise to the left bundle branch prior to entering the outflow tract. The axis itself was usually on the crest of the ventricular septum, but could be deviated leftward or rightward. Its proximity to the virtual basal plane reflected the angulation of the muscular septum. On average, the superior edge of the left bundle was within 3.3 mm of the hinge of the right coronary leaflet, with a range from 0.4 to 10.2 mm. The arrangement was markedly different in the case lacking an infero-septal recess. Our findings necessitated a redefinition of the right fibrous trigone and the central fibrous body. The atrioventricular conduction axis, having entered the aortic root, is usually closest at the hinge of the right coronary leaflet. Knowledge of the depth of the infero-septal recess, and the angulation of the muscular ventricular septal, may help to avoid conduction problems following transcatheter implantation of the aortic valve.

Similarities and differences in the arrangement of the atrioventricular conduction axis in the canine compared to the human heart.

BACKGROUND: Subtle differences exist between dog and human, despite use of the dog as a model for cardiac surgical and electrophysiological research. OBJECTIVE: The purpose of this study was to investigate the differences in the atrioventricular conduction axis and adjacent structures between dogs and humans. METHODS: We prepared 33 human and 5 canine hearts for serial histologic sections of the atrioventricular conduction axis, making correlations with gross anatomic findings. We additionally examined and photographed 15 intact normal human hearts obtained from infants undergoing autopsy. Furthermore, we interrogated a computed tomographic dataset from a human adolescent and from 2 autopsied canine hearts, both with normal cardiac anatomy. RESULTS: All canine hearts lacked an inferoseptal recess, with the noncoronary leaflet of the aortic valve and the right fibrous trigone having direct attachments to the septal surface of the left ventricular outflow tract. This correlated with an extensive nonbranching component of the ventricular conduction axis, which skirted half of the noncoronary aortic sinus. This anatomic arrangement was observed in 2 of 15 of autopsied infant hearts. In the human hearts with an inferoseptal recess, the relatively shorter nonbranching bundle is embedded within the fibrous tissue forming its right wall. CONCLUSION: We found a major difference between canine and the majority of human hearts, namely, the presence or absence of an inferoseptal recess. When this recess is absent, as in the canine heart and in some human hearts, a greater proportion of the atrioventricular conduction axis is found within the circumference of the subaortic outflow tract.

Cutaneous Leishmaniasis Caused by an Unknown Leishmania Strain, Arizona, USA.

We investigated an autochthonous case of cutaneous leishmaniasis caused by a genetically different Leishmania sp. in a patient in Arizona, USA. This parasite was classified into the subgenus Leishmania on the basis of multilocus DNA sequence and phylogenetic analyses of the rRNA locus and 11 reference genes.

Chromosome-Level Genome Sequence of Leishmania (Leishmania) tropica Strain CDC216-162, Isolated from an Afghanistan Clinical Case.

PacBio and Illumina MiSeq platforms were used for genomic sequencing of a Leishmania (Leishmania) tropica strain isolated from a patient infected in Pakistan. PacBio assemblies were generated using Flye v2.4 and polished with MiSeq data. The results represent a considerable improvement of the currently available genome sequences in the GenBank database.

Patent Macracanthorhynchus ingens Infection in a 17-Month-Old Child, Ohio.

Limited data exist on human Macracanthorhynchus infections. We report an asymptomatic 17-month-old who passed eggs and an adult Macracanthorhynchus ingens worm, indicating parasite maturation and reproduction. Macracanthorhynchus ingens may have a greater capacity to mature in humans versus Macracanthorhynchus hirudinaceus.

Three-dimensional visualization of the bovine cardiac conduction system and surrounding structures compared to the arrangements in the human heart.

In the human heart, the atrioventricular node is located toward the apex of the triangle of Koch, which is also at the apex of the inferior pyramidal space. It is adjacent to the atrioventricular portion of the membranous septum, through which it penetrates to become the atrioventricular bundle. Subsequent to its penetration, the conduction axis is located on the crest of the ventricular septum, sandwiched between the muscular septum and ventricular component of the membranous septum, where it gives rise to the ramifications of the left bundle branch. In contrast, the bovine conduction axis has a long non-branching component, which penetrates into a thick muscular atrioventricular septum having skirted the main cardiac bone and the rightward half of the non-coronary sinus of the aortic root. It commonly gives rise to both right and left bundle branches within the muscular ventricular septum. Unlike the situation in man, the left bundle branch is long and thin before it branches into its fascicles. These differences from the human heart, however, have yet to be shown in three-dimensions relative to the surrounding structures. We have now achieved this goal by injecting contrast material into the insulating sheaths that surround the conduction network, evaluating the results by subsequent computed tomography. The fibrous atrioventricular membranous septum of the human heart is replaced in the ox by the main cardiac bone and the muscular atrioventricular septum. The apex of the inferior pyramidal space, which in the bovine, as in the human, is related to the atrioventricular node, is placed inferiorly relative to the left ventricular outflow tract. The bovine atrioventricular conduction axis, therefore, originates from a node itself located inferiorly compared to the human arrangement. The axis must then skirt the non-coronary sinus of the aortic root prior to penetrating the thicker muscular ventricular septum, thus accounting for its long non-branching course. We envisage that our findings will further enhance comparative anatomical research.

The membranous septum revisited: A glimpse of our anatomical past.

The so-called membranous septum is the fibrous component of the septal structures within the heart. It is relatively subtle in its appearance, but of considerable significance to the understanding of cardiac function and cardiac disease, both congenital and acquired. Surprisingly, its existence was seemingly unknown until the early decades of the 19th century. At this time, those writing in the English language described it as the "undefended space," recognizing its importance in the setting of its aneurysmal dilation, and as the site of septal defects. By the initial decade of the 20th century, it had come to be recognized as the landmark to the site of atrioventricular bundle. Over the first decade of the 21st century, its clinical significance has been emphasized in the context of transcutaneous replacement of the aortic valve. In this review, we describe our own recent investigations of this fibrous part of the septal structures. At the same time, we provide a glimpse of our anatomic past, explaining how its initial description relied on the observations of young physicians taking their first steps in the investigation of cardiac anatomy.

Phytoremediation of polycyclic aromatic hydrocarbons (PAHs) in mangrove sediments using Rhizophora mangle.

A phytoremediation experiment was carried out in mesocosms to investigate the performance of Rhizophora mangle in the remediation of polycyclic aromatic hydrocarbons (PAHs) in mangrove sediment contaminated with crude oil. The water pH of the experiments (phytoremediation and natural attenuation) ranged from 4.9 to 8.4 at 0 and 90 days, respectively. The oxy-reduction potential (Eh) ranged from oxidising (108.0 mV, time 0) to reducing (approximately -110.0 mV, time 90) environments. Dissolved oxygen (DO) ranged from 5.7 mg L-1 (time 0) to 4.5 mg L-1 and 3.6 mg L-1 (time 90) in phytoremediation and natural attenuation, respectively. The sediments had silty texture and an average concentration of 5% organic matter (OM). Phytoremediation (60.76%) showed better efficiency in the remediation of the 16 PAHs compared to natural attenuation (49.57%). Principal component analyses showed a correlation between the concentrations of PAHs with pH, Eh, OM and DO in both experiments.

Physiological and Performance Impacts After Field Supramaximal High-Intensity Interval Training With Different Work-Recovery Duration.

High-intensity interval training (HIIT) has numerous external load control variables. The management of these variables makes the physiological responses and performance presented by athletes also modify. The present study aimed to assess the activity of CK and LDH enzymes, performance and metabolic responses caused by two HIIT protocols above the maximum in male recreational runners. Fifteen recreational male runners performed two HIIT protocols in randomized order with multiple conditions: 1) H15 (n = 15), with a HIIT protocol of 15:15 work-recovery duration, and 2) H30 (n = 15) with a HIIT protocol of 30:30 work-recovery duration. Both protocols were performed at similar intensity (130% vV̇O2 max ), one set until voluntary exhaustion. Blood samples were collected and used to capture the levels and activities of blood lactate (BLac: mmol⋅L-1), glucose (GLU: mg⋅dL-1), creatine kinase (CK: U⋅L-1), and lactate dehydrogenase (LDH: U⋅L-1). BLac and GLU were collected at pre, five, and ten minutes after the H15 and H30 protocols were performed. Blood samples were used to measure the activities of CK and LDH enzymes, which were verified 24 h before and 48 h after the protocols. The distance traveled (m), total time (s), and bouts performed (rep) were also registered. Significant differences between conditions H15 and H30 were observed in the bouts performed (p = 0.001; ES = 1.19). Several statistical differences were found over time for BLac [pre vs. post 5 (both conditions: p = 0.001), pre vs. post 10 (both conditions: p = 0.001), and post 5 vs. post 10 (H30: p = 0.004)], CK [pre vs. post 24 (H15: p < 0.001; ES = 0.97 and H30: p = 0.001; ES = 0.74) post 24 vs. post 48 (H30: p = 0.03; ES = 0.56)], and LDH [pre vs. post24 (H15: p = 0.008; ES = 1.07 and H30: p = 0.022; ES = 0.85). No statistical differences between conditions were observed for any blood parameter. Thus, the volunteers exhibited equal performance in both protocols, which resulted in a similar physiological response. Despite this similarity, in comparison to H15, the H30 protocol presented lower CK activity post 48 and lactate levels after 10 min post protocol.

Duplex Real-Time PCR Assay for Clinical Differentiation of Onchocerca lupi and Onchocerca volvulus.

In the United States and Europe, human onchocerciasis is a rare disease caused by zoonotic or anthropophilic parasites in the genus Onchocerca. The zoonotic species identified in focal areas of Europe and United States is Onchocerca lupi, and Onchocerca volvulus, the anthroponotic species, may be found among people who had lived in endemic areas of Africa, the Arabian Peninsula, or Latin America. Onchocerciasis due to O. lupi is an emergent parasitic disease, with limited diagnostic methods, in addition to the lack of information on its biology, transmission, and epidemiology. Cutaneous nodules are the disease's most prevalent manifestation but lack diagnostic specificity. To address the diagnosis of onchocerciasis at reference laboratories, we developed a duplex TaqMan real-time PCR (qPCR) method, targeting the cytochrome oxidase subunit I locus which has species-specific probes to identify and differentiate O. lupi from O. volvulus. We determined the performance of the duplex with a panel of 45 samples: 11 positives for O. lupi, six for O. volvulus, five samples with negative results for Onchocerca spp., and 23 non-Onchocerca nematodes. The duplex qPCR correctly detected 10 of 11 O. lupi- and six of six O. volvulus-positive specimens. The new duplex assay allowed the simultaneous detection and discrimination of O. lupi and O. volvulus in clinical specimens, expediting and facilitating the clinical diagnosis of O. lupi in non-endemic settings where the disease is an infrequent finding.

Leishmania infantum in US-Born Dog.

Leishmaniasis is a vectorborne disease that can infect humans, dogs, and other mammals. We identified one of its causative agents, Leishmania infantum, in a dog born in California, USA, demonstrating potential for autochthonous infections in this country. Our finding bolsters the need for improved leishmaniasis screening practices in the United States.

An Atypical Case of Autochthonous Cutaneous Leishmaniasis Associated with Naturally Infected Phlebotomine Sand Flies in Texas, United States.

In the United States, phlebotomine sand flies carrying Leishmania (Leishmania) mexicana are endemic along the southern border. However, relatively little is known about the enzootic and zoonotic transmission of L. (L.) mexicana within the United States, and autochthonous cases of the consequent disease are rarely reported. We investigated an atypical case of cutaneous leishmaniasis (CL) caused by L. (L.) mexicana in a patient from central Texas which did not respond to a typical antileishmanial chemotherapy. We also investigated sand fly vectors around the patient's residence. PCR followed by DNA sequencing was used for determination of Leishmania spp., sand fly species, and host blood meal source. The L. (L.) mexicana genotype from the patient was identical to one found in a positive sand fly. Moreover, this genotype presented the same single-nucleotide polymorphisms as other historical CL cases acquired in Texas over the last 10 years, but distinct from those originating in Mexico and Central America. Three sand fly species were identified among the samples analyzed (n = 194), the majority of which were Lutzomyia (Dampfomyia) anthophora (n = 190), of which four specimens tested positive for Leishmania and two blood-fed specimens showed the presence of a human blood meal. This study highlights the complexity of clinical management of CL in a setting where the disease is infrequently encountered. The detection of human blood in Lu. (D.) anthophora is the first documentation of anthropophagy in this species. This is the first report of wild-caught, naturally infected sand flies found in association with an autochthonous case of human leishmaniasis and the specific strain of Leishmania (Leishmania) mexicana in the United States.

Evaluation of accelerated biodegradation of oil-SPM aggregates (OSAs).

The studies of the formation of oil-Suspended Particulate Matter (SPM) aggregates (OSAs) have advanced significantly in the scientific community, however there is a need to accelerate oil biodegradation that was dispersed by the formation of OSAs. The present research presents a pioneering character regarding the addition of nutrients as biostimulus for autochthonous hydrocarboclastic bacteria in the biodegradation of Total Petroleum Hydrocarbons (TPH) dispersed by the formation of OSAs. Water aliquots were taken over 60 days from eight bioreactors to perform ionic species analysis, pH, salinity and temperature monitoring, liquid/liquid extraction, serial dilution methodology and filter membrane. TPH quantification was performed on the gas chromatograph with a flame ionisation detector (GC-FID). The addition of nutrients contributed positively to the rate and extent of biodegradation of TPH in association with field-collected SPM. The best result found was with the lowest nutrient concentration (Bio 1) with an average of 98.65% of TPH reduction.