Genome-wide SNP and microsatellite variation illuminate population-level epidemiology in the Leishmania donovani species complex

The species of the Leishmania donovani species complex cause visceral leishmaniasis, a debilitating infectious disease transmitted by sandflies. Understanding molecular changes associated with population structure in these parasites can help unravel their epidemiology and spread in humans. In this study, we used a panel of standard microsatellite loci and genome-wide SNPs to investigate population-level diversity in L. donovani strains recently isolated from a small geographic area spanning India, Bihar and Nepal, and compared their variation to that found in diverse strains of the L. donovani complex isolates from Europe, Africa and Asia. Microsatellites and SNPs could clearly resolve the phylogenetic relationships of the strains between continents, and microsatellite phylogenies indicated that certain older Indian strains were closely related to African strains. In the context of the anti-malaria spraying campaigns in the 1960s, this was consistent with a pattern of episodic population size contractions and clonal expansions in these parasites that was supported by population history simulations. In sharp contrast to the low resolution provided by microsatellites, SNPs retained a much more fine-scale resolution of population-level variability to the extent that they identified four different lineages from the same region one of which was more closely related to African and European strains than to Indian or Nepalese ones. Joining results of in vitro testing the antimonial drug sensitivity with the phylogenetic signals from the SNP data highlighted protein-level mutations revealing a distinct drug-resistant group of Nepalese and Indian L. donovani. This study demonstrates the power of genomic data for exploring parasite population structure. Furthermore, markers defining different genetic groups have been discovered that could potentially be applied to investigate drug resistance in clinical Leishmania strains.     

Leishmaniasis: vaccine candidates and perspectives

Leishmania is a protozoan parasite and a causative agent of the various clinical forms of leishmaniasis. High cost, resistance and toxic side effects of traditional drugs entail identification and development of therapeutic alternatives. The sound understanding of parasite biology is key for identifying novel drug targets, that can induce the cell mediated immunity (mainly CD4+ and CD8+ IFN-gamma mediated responses) polarized towards a Th1 response. These aspects are important in designing a new vaccine along with the consideration of the candidates with respect to their ability to raise memory response in order to improve the vaccine performance. This review is an effort to identify molecules according to their homology with the host and their ability to be used as potent vaccine candidates.     

Histamine H(1) and H(3) receptors in the rat thalamus and their modulation after systemic kainic acid administration

In rat thalamus, histamine H(1) receptor and isoforms of H(3) receptor were expressed predominantly in the midline and intralaminar areas. Correspondingly, higher H(1) and H(3) receptor binding was also detected in these areas. All isoforms of H(3) receptor were expressed in several thalamic nuclei, but there were minor differences between their expression patterns. H(1) mRNA expression was high in the ventral thalamus, but the H(1) binding level was low in these areas. Since increased brain histamine appears to have an antiepileptic effect through the H(1) receptor activity, kainic acid (KA)-induced status epilepticus in rat was used to study modulation of H(1) and H(3) receptors in the thalamus following seizures. After systemic KA administration, transient decreases in mRNA expression of H(1) receptor and H(3) receptor isoforms with full-length third intracellular loops were seen in the midline areas and the H(1) receptor mRNA expression also decreased in the ventral thalamus. After 1 week, a robust increase in mRNA expression of H(3) receptor isoforms with a full-length third intracellular loop was found in the ventral posterior, posterior, and geniculate nuclei. The changes indicate a modulatory role of H(3) receptor in the sensory and motor relays, and might be involved in possible neuroprotective and compensatory mechanisms after KA administration. However, short-term increases in the H(3) receptor binding appeared earlier (72 h) than the increases of H(3) mRNA expression (1-4 w). The elevations in H(3) binding were evident in the intralaminar area, laterodorsal, lateral posterior, posterior and geniculate nuclei, and were likely to be related to the cortical and subcortical inputs to thalamus.     

Single thalamic neurons which project to both the rostral cortex and caudate nucleus studied with the fluorescent double labeling method

After injections of fast blue into the rostral cortex and Evans blue into the caudate nucleus in cats, doubly labeled neurons were present in the ventral anterior, ventral lateral, rhomboid, and mediodorsal thalamic nuclei. Doubly labeled cells were also found in most members of the intralaminar group, including the central medial, paracentral, central lateral, and parafascicular nuclei. Although the centromedian nucleus contained large numbers of cells labeled with Evans blue which project to the caudate nucleus, and a few fast-blue labeled cells which projected to the cortex, doubly labeled neurons were absent from this posterior intralaminar nucleus in this study.     

Projections of the zona incerta in the cat, with stimulation controls.

In eight cats single electrolytic lesions were placed in the zona incerta, and resultant fiber degeneration studies were made. In seven additional cats, stimulating electrodes were chronically implanted bilaterally into the zona incerta, H₂ (lenticular fasciculus), or H (prerubral) fields of Forel. The animals were placed in a two-compartment shuttle box, and a routinely established procedure of subthalamic stimulation was instituted. When the sensory (nociceptive) or motor manifestations and reactions were established, small lesions were made through both poles of the electrodes. The brains were studied by silver techniques for degenerating axons and terminals. Findings in the latter group of animals with physiologic substrates, compared to those in the first group, indicated that the zona incerta contains at least two major physiologic-anatomic components with differential fiber projections. The first component is a medial zona incerta proper or caudalis, paleospinothalamic, nociceptive-conducting system which causes typical escape responses. Its unequivocal projections are to the nucleus of the H₁ field, posterior and dorsal hypothalamus, part of the intralaminar system, ventromedial and ventralis anterior nuclei, nucleus reuniens, reticular nucleus, pulvinar, posterior nucleus, central gray, red nucleus, and the central tegmental tract. The second constitutent concerns pyramidal-extrapyramidal motor type responses that arise with avoidance reactions from other portions of the zona incerta. In these cases there is heavy projection to the caudate, entopenduncular, globus pallidus, and putamen nuclei. In contrast, degeneration from the nociceptive part of the zona incerta or H₂ and H fields to these nuclei is minimal.     

Types of eye movements evoked by thalamic microstimulation in the alert cat

We characterized the type of eye movements evoked by electrical stimulation of the thalamic internal medullary lamina in alert cats. For comparison, other thalamic nuclei were studied: the lateral posterior, lateral dorsal, pulvinar complex, and ventral lateral nuclei. Three types of eye movements were distinguished: (i) Direction and amplitude-specific saccades were the most common type of eye movements evoked by electrical stimulation of the majority of the explored nuclei with the exception of the ventral lateral nucleus. Mean latency and threshold were 45 ms and 50 to 100 μA, compared with 60 ms and 200 to 300 μA for the same type of saccades evoked by electrical stimulation of the posterior thalamus. (ii) Saccades which appeared goal directed were evoked in 25% of the stimulations in the region of the internal medullary lamina. Their latency and threshold depended on the initial position of the eyes: The more deviated the eyes were from the “goal area,” the shorter the latency (minimum, 35 ms) and the lower the threshold (minimum, 16 μA). (iii) Centering saccades were evoked by stimulation of the ventral lateral nucleus. They were always accompanied by other movements. The minimum latency and threshold were, respectively, 90 ms and 200 μA. The finding that goal-directed saccades were selectively produced from certain internal medullary lamina sites is consistent with the view that a translation of spatial coordinates occurs at this level in the programming of eye movements.     

Pre- and post-alpha motoneuronal control of the soleus H-reflex during sinusoidal hip movements in human spinal cord injury

The aim of this study was to establish the contribution of hip-mediated sensory feedback to spinal interneuronal circuits during dynamic conditions in people with incomplete spinal cord injury (SCI). Specifically, we investigated the effects of synergistic and antagonistic group I afferents on the soleus H-reflex during imposed sinusoidal hip movements. The soleus H-reflex was conditioned by stimulating the common peroneal nerve (CPN) at short (2, 3, and 4 ms) and long (80, 100, and 120 ms) conditioning test (C-T) intervals to assess the reciprocal and pre-synaptic inhibition of the soleus H-reflex, respectively. The soleus H-reflex was also conditioned by medial gastrocnemius (MG) nerve stimulation at C-T intervals ranging from 4 to 7 ms to assess changes in autogenic Ib inhibition during hip movement. Sinusoidal hip movements were imposed to the right hip joint at 0.2 Hz by the Biodex system while subjects were supine. The effects of sinusoidal hip movement on five leg muscles along with hip, knee, and ankle joint torques were also established during sensorimotor conditioning of the reflex. Phase-dependent modulation of antagonistic and synergistic muscle afferents was present during hip movement, with the reciprocal, pre-synaptic, and Ib inhibition to be significantly reduced during hip extension and reinforced during hip flexion. Reflexive muscle and joint torque responses--induced by the hip movement--were entrained to specific phases of hip movement. This study provides evidence that hip-mediated input acts as a controlling signal of pre- and post-alpha motoneuronal control of the soleus H-reflex. The expression of these spinal interneuronal circuits during imposed sinusoidal hip movements is discussed with respect to motor recovery in humans after SCI.     

Infectious diseases in paediatric pathology: experience from a developing country

Infectious and parasitic diseases have always challenged man. Although many of them are typically seen in some areas of the world and can be adequately managed by just improving socioeconomic status and sanitary conditions, they are still quite prevalent and may sometimes be seen outside their original geographical areas. Human migration due to different reasons, tourism, blood transfusion and solid organ transplantation has created new concerns for health professionals all over the world. If not for diagnostic purposes, at least these tropical and infectious diseases should be largely known because their epidemiology, pathogenesis, host/parasite interaction, inflammatory and reparative responses are quite interesting and teach us about human biology. Curiosity is inherent to pathology practice and so we are compelled to look for things other than tumours or degenerative diseases. This review focuses on infectious and parasitic diseases found in a developing country and brings up-to-date information on diseases caused by viruses (dengue, yellow fever), bacteria (typhoid fever, leprosy), parasites (Chagas' disease, cutaneous and visceral leishmaniasis, amoebiasis, Capillaria hepatica, schistosomiasis, cysticercosis) and caused by fungi (paracoccidioidomycosis, cryptococcosis, histoplasmosis) that may be useful for pathologists when facing somewhat strange cases from developing countries.     

Application of Fractal Dimension and Texture Analysis to Evaluate the Effectiveness of Treatment of a Venous Lake in the Oral Mucosa Using a 980 nm Diode Laser—A Preliminary Study

Venous lake (VL) is a common lesion resulting from the enlargement of thin-walled veins surrounded by a thick wall of fibrous tissue. Photocoagulation is becoming one of the basic methods for treating small vascular lesions. The aim of this study was an application of texture analysis (TA) and fractal dimension analysis (FDA) to evaluate VL treatment. Twenty-three VLs were included in the study. We used a 980 nm diode laser, 6 W, 100 ms pulse mode with a 50% duty cycle. The total dose of energy was in a range from 80 J to 600 J. We used the difference of intensity algorithm for FDA and microcontrast and a co-occurrence matrix for TA. A complete therapeutic effect was achieved in 83%, and in 9%, scar formation was observed after three months. No healing was observed in 4%, and there was partial healing in 4%. No differences in FD were observed between the lesions after three months and the healthy mucosa. The fractal dimension and microcontrast of a vascular lesion are mutually coupled. FDA and TA is a useful and objective method of assessing treatment effects for venous lakes. The non-contact mode of the 980 nm diode laser is an effective and safe method for treating a VL.