Morphological and cytochemical characterization of the peripheral blood cells of farmed streaked prochilod Prochilodus lineatus (Characiformes, Prochilodontidae)

ABSTRACT Morphological and cytochemical studies of peripheral blood cells of fish have improved the understanding of their functions and cell types. The present study performed the Morphological and cytochemical analysis of the peripheral blood of Prochilodus lineatus, Characiform native to South America, which has been gaining space in local aquaculture and as a species introduced in Asia. Our analysis provided information on the morphological and cytochemical characteristics of the leukocytes, for the formulation of hypothesis about their role in the immune system of the species. It was found that Prochilodus lineatus has morphological and cytochemical features in common with other fish species, mainly of the Characiformes order. However, we detected the presence of heterophils and PAS positive granulocytes simultaneously with neutrophils. We also found that heterophils and PAS positive granulocytes are very similar, both morphologically and cytochemically.

Diagnostic Utility of Cerebrospinal Fluid Procalcitonin in Neonatal Meningitis

We aimed to study the diagnostic utility of cerebrospinal fluid (CSF) procalcitonin in neonates with meningitis. All the neonates with sepsis who qualified for lumbar puncture were prospectively evaluated. The neonates were classified as Meningitis and No meningitis group based on predefined criteria. CSF procalcitonin was estimated in these neonates along with cytological and biochemical parameters. A total of 113 neonates were included in the study with 29 in meningitis group and 84 in no meningitis group. The median procalcitonin levels was higher in babies with meningitis as compared to those without meningitis [0.194 (0.034 - 0.534) in meningitis group vs 0.012 (0.012-0.012) ng/ml in no meningitis group, p< 0.001]. The area under curve (AUC) for CSF procalcitonin was 0.867 (0.77 -0.95) and at a cut off level of 0.120 ng/ml CSF procalcitonin had a sensitivity of 83%, specificity of 84% and positive and negative predictive likelihood ratios of 5.35 and 0.20 respectively for the diagnosis of meningitis.Conclusion: CSF procalcitonin has a good diagnostic accuracy similar to other parameters in the diagnosis of neonatal meningitis and can be considered as an additional diagnostic marker particularly, when CSF culture is negative and cytochemical analysis is inconclusive.Trial registration number and date: CTRI/2018/09/015720 ; 14/09/2018

Cryptococcal Meningitis in an HIV-Negative Puerperal Woman

Cryptococcal meningitis is a common opportunistic infection in HIV-infected patients and other immunocompromised people. Pregnancy, which is a state of relative immunosuppression, can also be a risk factor for the development of cryptococcal meningitis. We report a clinical case of a 41-year-old woman who developed a severe meningeal syndrome after an otherwise normal pregnancy. Cerebrospinal fluid (CSF) cytochemical analysis presented hypoglycorrhachia, high protein levels, and pleocytosis. Cryptococcal antigen tested positive in serum and CSF, and Cryptococcus neoformans was identified in the CSF culture. The diagnosis of cryptococcal meningitis was confirmed, and antifungal induction therapy was started with liposomal amphotericin B and flucytosine. After clinical improvement, induction therapy was discontinued, and the patient was discharged under maintenance therapy with fluconazole. While under antifungal maintenance therapy, the patient presented worsening of symptoms and a new brain magnetic resonance showed the development of multiple cryptococcoma. Despite sterile CSF cultures, there was a deterioration of the cytochemical parameters. The diagnosis of immune reconstitution inflammatory syndrome was assumed, and after initiation of corticotherapy, the patient improved considerably. This is a rare case of cryptococcal meningitis in a puerperal woman with a challenging management.

LeishIF4E-5 Is a Promastigote-Specific Cap-Binding Protein in Leishmania

Leishmania parasites cycle between sand fly vectors and mammalian hosts, transforming from extracellular promastigotes that reside in the vectors’ alimentary canal to obligatory intracellular non-motile amastigotes that are harbored by macrophages of the mammalian hosts. The transition between vector and host exposes them to a broad range of environmental conditions that induces a developmental program of gene expression, with translation regulation playing a key role. The Leishmania genome encodes six paralogs of the cap-binding protein eIF4E. All six isoforms show a relatively low degree of conservation with eIF4Es of other eukaryotes, as well as among themselves. This variability could suggest that they have been assigned discrete roles that could contribute to their survival under the changing environmental conditions. Here, we describe LeishIF4E-5, a LeishIF4E paralog. Despite the low sequence conservation observed between LeishIF4E-5 and other LeishIF4Es, the three aromatic residues in its cap-binding pocket are conserved, in accordance with its cap-binding activity. However, the cap-binding activity of LeishIF4E-5 is restricted to the promastigote life form and not observed in amastigotes. The overexpression of LeishIF4E-5 shows a decline in cell proliferation and an overall reduction in global translation. Immuno-cytochemical analysis shows that LeishIF4E-5 is localized in the cytoplasm, with a non-uniform distribution. Mass spectrometry analysis of proteins that co-purify with LeishIF4E-5 highlighted proteins involved in RNA metabolism, along with two LeishIF4G paralogs, LeishIF4G-1 and LeishIF4G-2. These vary in their conserved eIF4E binding motif, possibly suggesting that they can form different complexes.

Bioregulatory approach to stimulating sport horses

The presented work presents the results of the effectiveness of the bioregulatory approach in stimulating sports horses. The aim of the study was to find a way to safely restore the energy balance and prevent the occurrence of inflammatory processes in the musculoskeletal system during significant physical exertion. Sport horses spend a lot of energy not only in competition, but also during daily training. Such loads require a high supply of energy. Often the body uses its last reserves to compensate for its energy needs, which leads to exhaustion and the appearance of various diseases. First of all, diseases occur in the musculoskeletal system, because it is the limbs that bear the greatest load during training. As you know, various synthetic doping is introduced to keep animals in a sporty tone. Their action is not long-lasting, but it has many side effects for the animal's body and leads to disruption of the functioning of organs and systems. Today, there is an alternative to such stimulation methods thanks to a bioregulatory approach in the treatment of animals using preparations containing exclusively natural ingredients. To confirm the effectiveness of the use of bioregulatory drugs, clinical and laboratory studies of animal blood (general, biochemical, cytochemical) were carried out. The main changes in laboratory parameters were obtained by cytochemical analysis of such enzymes as GPDH, SDH. These enzymes are among the main indicators of the intensity of energy metabolism in the horse's body. The quantitative composition of basophils and eosinophils has also changed, which indicates a decrease in the level of the inflammatory process in the body of experimental animals. The results obtained prove the effectiveness of the investigated drugs and their direct influence on the energy balance of the “horse-athlete”. Thanks to the natural composition of the components of the preparations used, side effects on the body can be avoided and used without restrictions throughout the horse's sports career.

Activity of myeloperoxidase of neutrophils in the blood of pregnant women with diffuse toxic goiter

Aim. To study the activity of myeloperoxidase in the peripheral blood of pregnant women in normal and thyroid diseases. Materials and methods. Cytochemical analysis was used to study the activity of myeloperoxidase in peripheral blood neutrophils of pregnant women with diffuse toxic goiter, as well as in women with the physiological course of pregnancy in a comparative aspect. Results. It was found that in women with a physiological course of pregnancy, the cytochemical parameters of myeloperoxidase in neutrophils were reduced only in the 1st trimester of pregnancy. In patients with diffuse toxic goiter, with severe thyrotoxicosis syndrome, the activity of the enzyme was significantly reduced during all periods of pregnancy. Apparently, these shifts of the enzyme, which is the main component of the protective system of blood phagocytes, are associated with their functional stress. Conclusion. Based on this opinion, it seems that blood neutrophils enter the system of thyroid hormone metabolism by their degradation. Apparently, this mechanism is activated with an increased content of hormones in the circulation, which occurs during pregnancy, and especially with concomitant diffuse toxic goiter with thyrotoxicosis syndrome and is aimed at preserving homeostasis necessary for normal fetal development.

PHOSPHATASE ACTIVITY AND POSSIBILITIES OF LYSOSOMAL SECRETION OF AIRWAY NEUTROPHILS IN PATIENTS WITH BRONCHIAL ASTHMA IN COLD-INDUCED STRESS

An approach to the study of cellular inflammation using methods of cytochemical analysis of sputum in patients with bronchial asthma with different types of airway reactions to bronchoprovocation with cold air is presented. Endotyping of patients contributes to a better understanding of the pathophysiological mechanisms and the choice of treatment tactics for the disease.

NaCl-altered oxygen flux profiles and H+-ATPase activity in roots of two contrasting poplar species

Maintaining mitochondrial respiration is crucial for proving ATP for H+ pumps to continuously exclude Na+ under salt stress. NaCl-altered O2 uptake, mitochondrial respiration, and the relevance to H+-ATPase activity were investigated in two contrasting poplar species, Populus euphratica (salt-tolerant) and P. popularis 35–44 (salt-sensitive). Compared with P. popularis, P. euphratica roots exhibited a greater capacity to extrude Na+ under NaCl stress (150 mM). The cytochemical analysis with Pb(NO3)2 staining revealed that P. euphratica root cells retained higher H+ hydrolysis activity than the salt-sensitive poplar during a long-term (LT) of increasing salt stress (50 to 200 mM NaCl, 4 weeks). Long-sustained activation of proton pumps require long-lasting supply of energy (ATP), delivered by aerobic respiration. Taking advantage of the vibrating-electrodes technology combined with the use of membrane-tipped, polarographic oxygen microelectrodes, the species, spatial, and temporal differences in root O2 uptake were characterized under conditions of salt stress. Oxygen uptake upon NaCl shock (150 mM) was less declined in P. euphratica than in P. popularis, although the salt-induced transient kinetics were distinct from the drastic drop of O2 caused by hyperosmotic shock (255 mM mannitol). Short-term (ST) treatment (150 mM NaCl, 24 h) stimulated O2 influx in P. euphratica roots, and LT-treated P. euphratica displayed an increased O2 influx along root axis, whereas O2 influx declined with increasing salinity in P. popularis roots. The spatial localization of O2 influxes revealed that the apical zone was more susceptible than elongation region upon high NaCl (150, 200 mM) during ST and LT stress. Pharmacological experiments showed that the Na+ extrusion and H+-ATPase activity in salinized roots were correspondingly suppressed when O2 uptake was inhibited by a mitochondrial respiration inhibitor, NaN3. Therefore, we conclude that the stable mitochondrial respiration energized H+-ATPase of P. euphratica root cells for maintaining Na+ homeostasis under salt environments.