Immature Forms Research Articles

Aedes aegypti surveillance under the International Health Regulations - experience at Madeira Island

Abstract Problem Aedes aegypti was first identified on Madeira Island in 2005, persisting since then. This mosquito transmits various haemorrhagic diseases and caused a Dengue outbreak in 2012-2013. The island has two seaports and one airport that may serve as points of entry or dissemination of these diseases, potentially leading to a new outbreak or international disease spread. We aimed to assess Ae. aegypti presence at the island’s seaports and airport during 2023. Description Data was collected from 52 entomological panels produced by the Regional Health Department throughout 2023. This data is publicly available on the Institute of Health Administration’s website. Ovitraps capture eggs, larvae, and pupae of Ae. aegypti, and BGtraps capture adult mosquitos. Positivity refers to the proportion of times an ovitrap recorded eggs in the entomological cycle. Results In 2023, there were 204 active ovitraps at the island (17 at the airport, 9 at Funchal’s seaport 8 at Caniçal’s seaport) and 24 BGtraps (2 at the airport, 2 at Funchal’s port and 1 at Caniçal’s port). Weekly reports revealed a median of 2243 (0-14448) eggs and 14 (0-234) mosquitos in all traps. Positivity was higher in the second semester. On average, ovitraps’ positivity at the airport (avg 6.4%; 0-59%) and at Caniçal’s port (avg 2.8%; 0-47%) were lower than the positivity of the island (avg 14.8%). Funchal’s port had a higher positivity than the island (avg 15.9%; 0-67%). Per-ovitrap, the average weekly positivity rate rose by 15%, 10% and 5% at Caniçal’s port, airport and Funchal’s port, respectively, compared to 2012-2022. Lessons Ae. aegypti’s eggs prevalence increased in 2023. The high positivity of eggs suggests an increased likelihood of mosquitos nearby. The detection of immature forms triggers actions to prospect the surrounding area for identification and elimination of breeding grounds, as well as the placement of salt in water accumulation areas. This is particularly important at high-traffic areas such as seaports and airports. Key messages • The increase in Ae. aegypti egg positivity in 2023 on Madeira Island suggests a higher risk of disease transmission. • It is essential to prioritize vector control measures at seaports and the airport in Madeira Island.

Cannabis Use and Cannabidiol Modulate HIV-Induced Alterations in TREM2 Expression: Implications for Age-Related Neuropathogenesis.

Triggering receptor expressed on myeloid cells 2 (TREM2) is involved in neuroinflammation and HIV-associated neurocognitive impairment (NCI). People with HIV (PWH) using cannabis exhibit lower inflammation and neurological disorders. We hypothesized that TREM2 dysfunction mediates HIV neuropathogenesis and can be reversed by cannabinoids. EcoHIV-infected wildtype (WT) and TREM2R47H mutant mice were used to study HIV's impact on TREM2 and behavior. TREM2 and related gene expressions were examined in monocyte-derived macrophages (MDMs) from PWH (n = 42) and people without HIV (PWoH; n = 19) with varying cannabis use via RNA sequencing and qPCR. Differences in membrane-bound and soluble TREM2 (sTREM2) were evaluated using immunocytochemistry (ICC) and ELISA. EcoHIV increased immature and C-terminal fragment forms of TREM2 in WT mice but not in TREM2R47H mice, with increased IBA1 protein in TREM2R47H hippocampi, correlating with worse memory test performance. TREM2 mRNA levels increased with age in PWoH but not in PWH. Cannabidiol (CBD) treatment increased TREM2 mRNA alone and with IL1β. RNA-seq showed the upregulation of TREM2-related transcripts in cannabis-using PWH compared to naïve controls. IL1β increased sTREM2 and reduced membrane-bound TREM2, effects partially reversed by CBD. These findings suggest HIV affects TREM2 expression modulated by cannabis and CBD, offering insights for therapeutic strategies.

Abstract A015: Tissue MUC5AC expression predicts recurrence patterns following resection in pancreatic ductal adenocarcinoma

Abstract Pancreatic ductal adenocarcinoma (PDA) has consistently poor outcomes, even for early-stage tumors that undergo resection (>70%), despite completion of prescribed perioperative systemic therapy. Mucin 5AC (MUC5AC), a heavy glycoprotein, exists in two major glycoforms: heavily glycosylated mature (MM) and less glycosylated immature (IM) forms. MM is located in the apical region and extracellular space, whereas IM is in the perinuclear region. Our previous work demonstrated the association between MM and treatment response and outcomes in resected PDA after neoadjuvant therapy (NAT). In this study, we investigate the impact of MUC5AC tissue expression in resected PDA samples on recurrence site characteristics (local (LR) vs. distant metastasis (mets) vs. both LR and mets). Resected PDA formalin-fixed paraffin-embedded tissue blocks from January 2010 to June 2021 were obtained from the Ohio State University biorepository. Immunohistochemistry was performed to study the expression for MM and IM using monoclonal antibodies 45M1 and CLH2, respectively, and H-scores (0-300) were calculated. Logistic regression was used to study this association of interest. We had 100 R-PDA (43 received NAT, and 57 had upfront surgery (UpS)) available for testing. The median age of diagnosis was 65 years, and 50% were males. Most of the patients in the NAT group received FOLFIRINOX (n=36) and received gemcitabine/nab-paclitaxel (n=5) or FOLFOX (n=2). The median number of doses received is 6 (range 2-9). The results in these groups can be summarized as follows: a) 35/43 had recurrence (LR=7, mets=21, and LR+mets=7), with the liver being the most common site of metastasis (13/28) followed by lung (5/28) and peritoneum (5/28), b) Higher IM expression (mean H-score) was significantly (p=0.0275) higher in LR+mets (H-score 196) than in LR (H-score 69) or mets (H-score 115) groups, c) Higher MM expression (mean H-score) was also significantly different among these groups (LR vs. mets vs. LR+mets = 81 vs. 120 vs. 206, p= 0.0358), e) similar trend of mean H-scores was noted in FOLFIRINOX sub-group (LR vs. mets vs. LR+mets = 205 vs. 109 vs. 60, p=0.0185 for IM and 65 vs. 116 vs. 205, p=0.0329 for MM). No significant association between recurrence patterns and tissue MUC5AC expression in the UpS group. These findings concur with our prior observations, underscoring the clinical significance of MUC5AC in resected PDA post-NAT and suggesting its potential role in facilitating pancreatic tumor cell metastasis. Although larger prospective studies are needed to confirm these results, our study highlights MUC5AC as a potential therapeutic target to prevent metastasis in early-stage PDA, potentially improving overall outcomes. Citation Format: Ashish Manne, Lianbo Yu, Ashwini Esnakula. Tissue MUC5AC expression predicts recurrence patterns following resection in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A015.

Antibacterial Activity and Prebiotic Properties of Six Types of Lamiaceae Honey.

Our work investigated the antimicrobial and prebiotic properties of basil, mint, oregano, rosemary, savory, and thyme honey. The potential antimicrobial action, assessed against the pathogens Acinetobacter baumannii, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus, evidenced the capacity of the honey to influence the pathogenic hydrophobicity and hemolytic activities. Honey inhibited pathogen biofilms, acting especially on the mature biofilms, with inhibition rates of up to 81.62% (caused by the presence of mint honey on L. monocytogenes). S. aureus biofilms were the most susceptible to the presence of honey, with inhibition rates up of to 67.38% in the immature form (caused by basil honey) and up to 80.32% in the mature form (caused by mint honey). In some cases, the amount of nuclear and proteic material, evaluated by spectrophotometric readings, if also related to the honey's biofilm inhibitory activity, let us hypothesize a defective capacity of building the biofilm scaffold or bacterial membrane damage or an incapability of producing them for the biofilm scaffold. The prebiotic potentiality of the honey was assessed on Lacticaseibacillus casei Shirota, Lactobacillus gasseri, Lacticaseibacillus paracasei subsp. paracasei, and Lacticaseibacillus rhamnosus and indicated their capacity to affect the whole probiotic growth and in vitro adhesive capacity, as well as the antioxidant and cytotoxic abilities, and to inhibit, mainly in the test performed with the L. casei Shirota, L. gasseri, and L. paracasei supernatants, the immature biofilm of the pathogens mentioned above.

Effects of the El Niño-Southern Oscillation and seasonal weather conditions on Aedes aegypti infestation in the State of São Paulo (Brazil): A Bayesian spatio-temporal study.

Seasonal fluctuations in weather are recognized as factors that affect both Aedes (Ae.) aegypti mosquitoes and the diseases they carry, such as dengue fever. The El Niño-Southern Oscillation (ENSO) is widely regarded as one of the most impactful atmospheric phenomena on Earth, characterized by the interplay of shifting ocean temperatures, trade wind intensity, and atmospheric pressure, resulting in extensive alterations in climate conditions. In this study, we investigate the influence of ENSO and local weather conditions on the spatio-temporal variability of Ae. aegypti infestation index. We collected seasonal entomological survey data of immature forms of Ae. aegypti mosquitoes (Breteau index), as well as data on temperature, rainfall and the Oceanic Niño Index (ONI) for the period 2008-2018 over the 645 municipalities of the subtropical State of São Paulo (Brazil). We grounded our analytical approach on a Bayesian framework and we used a hierarchical spatio-temporal model to study the relationship between ENSO tracked by ONI, seasonal weather fluctuations and the larval index, while adjusting for population density and wealth inequalities. Our results showed a relevant positive effect for El Niño on the Ae. aegypti larval index. In particular, we found that the number of positive containers would be expected to increase by 1.30-unit (95% Credible Intervals (CI): 1.23 to 1.37) with El Niño events (i.e., ≥ 1°C, moderate to strong) respect to neutral (and weak) events. We also found that seasonal rainfall exceeding 153.12 mm appears to have a notable impact on vector index, leading potentially to the accumulation of ample water in outdoor discarded receptacles, supporting the aquatic phase of mosquito development. Additionally, seasonal temperature above 23.30°C was found positively associated to the larval index. Although the State of São Paulo as a whole has characteristics favourable to proliferation of the vector, there were specific areas with a greater tendency for mosquito infestation, since the most vulnerable areas are predominantly situated in the central and northern regions of the state, with hot spots of abundance in the south, especially during El Niño events. Our findings also indicate that social disparities present in the municipalities contributes to Ae. aegypti proliferation. Considering the anticipated rise in both the frequency and intensity of El Niño events in the forthcoming decades as a consequence of climate change, the urgency to enhance our ability to track and diminish arbovirus outbreaks is crucial.

The FLARE Score and Circulating Neutrophils in Patients with Cancer and COVID-19 Disease.

Inflammation and neutrophils play a central role in both COVID-19 disease and cancer. We aimed to assess the impact of pre-existing tumor-related inflammation on COVID-19 outcomes in patients with cancer and to elucidate the role of circulating neutrophil subpopulations. We conducted a multicenter retrospective analysis of 524 patients with cancer and SARS-CoV-2 infection, assessing the relationship between clinical outcomes and circulating inflammatory biomarkers collected before and during COVID-19 infection. Additionally, a single-center prospective cohort study provided data for an exploratory analysis, assessing the immunophenotype of circulating neutrophils and inflammatory cytokines. The primary endpoints were 30-day mortality and the severity of COVID-19 disease. Prior to COVID-19, 25% of patients with cancer exhibited elevated dNLR, which increased to 55% at the time of COVID-19 diagnosis. We developed the FLARE score, incorporating both tumor- and infection-induced inflammation, which categorized patients into four prognostic groups. The poor prognostic group had a 30-day mortality rate of 68%, significantly higher than the 23% in the favorable group (p < 0.0001). This score proved to be an independent predictor of early mortality. This prospective analysis revealed a shift towards immature forms of neutrophils and higher IL-6 levels in patients with cancer and severe COVID-19 infection. A pre-existing tumor-induced pro-inflammatory state significantly impacts COVID-19 outcomes in patients with cancer. The FLARE score, derived from circulating inflammatory markers, emerges as an easy-to-use, globally accessible, effective tool for clinicians to identify patients with cancer at heightened risk of severe COVID-19 complications and early mortality who might benefit most from immediate and intensive treatment strategies. Furthermore, our findings underscore the significance of immature neutrophils in the progression of COVID-19 in patients with cancer, advocating for further investigation into how these cells contribute to both cancer and COVID-19 disease.

Molecular identification and genetic variability of equine and bovine ocular setariasis in india: molecular profiling by mitochondrial genes.

Ocular setariasis is an ectopic infection caused by a parasite under the genus Setaria. Adult worms belong to the Setariidae family and typically reside in the peritoneal cavity of ungulates. However, immature forms of these species may aberrantly migrate to the eyes of cattle, buffalo, goats, horses and several other hosts, leading to corneal opacity and blindness. Here, we have distinguished the Setaria digitata collected from both equine and buffalo hosts based on the morphology, molecular profiling of mitochondrial cytochrome c oxidase subunit 1 (Cox1), cytochrome c oxidase subunit 3 (Cox3) and, Nicotinamide Adenine Dinucleotide dehydrogenase subunit 1 (NAD1) genes. A single filarial worm was collected from the eye of one equine and one bovine host. These worms were then processed for morphological examination and DNA isolation. Cox1, Cox3 and NAD1 genes were amplified using specific primers and subjected to custom sequencing. The sequences were then used for multiple sequence alignment, assessment of entropy, similarity and haplotype diversity analysis. Key morphological features confirmed the worms collected were male and female Setaria digitata from equine and buffalo hosts, respectively. Cox1, Cox3 and NAD1 gene sequence analysis showed a close association of S.digitata Indian isolates with its counterparts from Sri Lanka and China isolates. The phylogram of bovine S. digitata sequences shows a close relationship to other equine S. digitata sequences, indicating the need for further in-depth studies on the prevalence of infection across various host species and intermediate hosts. Although the sequence results suggest that S. digitata is likely the causative agent of ocular setariasis in India, additional samples are needed to confirm this conclusion. Comprehensive analysis of the transcriptome and proteome of S. digitata from both bovine and equine hosts is necessary to explore variations in host-parasite interactions. These findings will aid in future parasite identification, investigations into vector prevalence in India, and the development of control measures against ocular setariasis.

Atomic resolution map of the solvent interactions driving SOD1 unfolding in CAPRIN1 condensates

Biomolecules can be sequestered into membrane-less compartments, referred to as biomolecular condensates. Experimental and computational methods have helped define the physical-chemical properties of condensates. Less is known about how the high macromolecule concentrations in condensed phases contribute "solvent" interactions that can remodel the free-energy landscape of other condensate-resident proteins, altering thermally accessible conformations and, in turn, modulating function. Here, we use solution NMR spectroscopy to obtain atomic resolution insights into the interactions between the immature form of superoxide dismutase 1 (SOD1), which can mislocalize and aggregate in stress granules, and the RNA-binding protein CAPRIN1, a component of stress granules. NMR studies of CAPRIN1:SOD1 interactions, focused on both unfolded and folded SOD1 states in mixed phase and demixed CAPRIN1-based condensates, establish that CAPRIN1 shifts the SOD1 folding equilibrium toward the unfolded state through preferential interactions with the unfolded ensemble, with little change to the structure of the folded conformation. Key contacts between CAPRIN1 and the H80-H120 region of unfolded SOD1 are identified, as well as SOD1 interaction sites near both the arginine-rich and aromatic-rich regions of CAPRIN1. Unfolding of immature SOD1 in the CAPRIN1 condensed phase is shown to be coupled to aggregation, while a more stable zinc-bound, dimeric form of SOD1 is less susceptible to unfolding when solvated by CAPRIN1. Our work underscores the impact of the condensate solvent environment on the conformational states of resident proteins and supports the hypothesis that ALS mutations that decrease metal binding or dimerization function as drivers of aggregation in condensates.

Tertiary Lymphoid Structures in Brain Metastases of Lung Cancer: Prognostic Significance and Correlation With Clinical Outcomes.

The prognosis of patients with brain metastases (BMs) originating from lung cancer remains poor, despite advancements in treatment strategies. The role of tertiary lymphoid structures (TLSs) within the tumor immune microenvironment of BMs has not been extensively explored. This study utilized patient-derived clinical samples from 17 patients with histologically confirmed BMs of lung cancer, undergoing surgical resection. Immunohistochemistry was employed to analyze the presence and characteristics of TLS and tumor-infiltrating lymphocytes (TILs) within BM tissues, correlating these with clinical outcomes. TLSs, albeit in their immature form, were identified within BM tissues, distinguishing them from their mature counterparts in primary lung cancer tissues. A significant correlation between TLS density (but not TIL density) and improved postoperative survival was observed, underscoring the potential of TLS density as an independent prognostic marker. Furthermore, TLS density did not correlate with the Graded Prognostic Assessment (GPA) index, suggesting its unique prognostic value beyond conventional predictors. Our findings reveal the presence of TLSs in lung cancer-derived BMs and highlight their prognostic significance, independent of the GPA index. The identification of TLS within the unique central nervous system tumor microenvironment offers new insights into the immune landscape of BMs and suggests potential avenues for immunotherapeutic interventions targeting these structures to improve patient outcomes.

Prevalence of Bovine Fasciolosis with Fasciola Hepatica in the M'pila Slaughter Areas of Congo Brazzaville

Fasciolosis is a hepatobiliary dystomatosis due to the migration into the liver parenchyma of immature form of parasites called fasciola hepatica or liver fluke. It is a cosmopolitan disease which represents economic and health problems whose diagnosis is rarely made, it is usually a discovery at the slaughterhouse. It mainly affects animals and sometimes humans. It is in this context that this study was carried out at two slaughterhouses in Mpila in order to determine the prevalence of bovine fasciolosis according to breeds, age, sex, origin and slaughter sector. The liver incision technique to search for flukes was carried out on 356 cattle. The results of the analysis carried out on 356 cattle including 134 infested cattle showed a prevalence of 37.64%. Furthermore, the highest prevalence was observed among mixed race people, a rate of 53.84%. Thus, cattle aged over 4 years were the most exposed to this pathology. Our results show that fasciolosis is a reality in the slaughter areas of Brazzaville. Monitoring of this pathology must be carried out to protect the health of our animals and that of humans.

A sperm-activating trypsin-like protease from the male reproductive tract of Spodoptera litura: Proteomic identification, sequence characterization, gene expression profile, RNAi and the effects of ionizing radiation

Like other lepidopteran insects, males of the tobacco cutworm moth, Spodoptera litura produce two kinds of spermatozoa, eupyrene (nucleate) and apyrene (anucleate) sperm. Formed in the testis, both kinds of sperm are released into the male reproductive tract in an immature form and are stored in the duplex region of the tract. Neither type of sperm is motile at this stage. When stored apyrene sperm from the duplex are treated in vitro with an extract of the prostatic region of the male tract, or with mammalian trypsin, they become motile; activation is greater and achieved more rapidly with increasing concentration of extract or enzyme. The activating effect of prostatic extract is blocked by soybean trypsin inhibitor (SBTI), also in a dose-dependent way. These results suggest that the normal sperm-activating process is due to an endogenous trypsin-like protease produced in the prostatic region.Proteomic analysis of S. litura prostatic extracts revealed a Trypsin-Like Serine Protease, TLSP, molecular weight 27 kDa, whose 199-residue amino acid sequence is identical to that of a predicted protein from the S. litura genome and is highly similar to predicted proteins encoded by genes in the genomes of several other noctuid moth species. Surprisingly, TLSP is only distantly related to Serine Protease 2 (initiatorin) of the silkmoth, Bombyx mori, the only identified lepidopteran protein so far shown to activate sperm. TLSP has features typical of secreted proteins, probably being synthesized as an inactive precursor zymogen, which is later activated by proteolytic cleavage.cDNA was synthesized from total RNA extracted from the prostatic region and was used to examine TLSP expression using qPCR. tlsp mRNA was expressed in both the prostatic region and the accessory glands of the male tract. Injection of TLSP-specific dsRNA into adult males caused a significant reduction after 24 h in tlsp mRNA levels in both locations. The number of eggs laid by females mated to adult males that were given TLSP dsRNA in 10 % honey solution, and the fertility (% hatched) of the eggs were reduced. Injecting pupae with TLSP dsRNA caused the later activation of apyrene sperm motility by adult male prostatic extracts to be significantly reduced compared to controls.Exposure of S. litura pupae to ionizing radiation significantly reduced expression of tlsp mRNA in the prostatic part and accessory gland of irradiated males in both the irradiated generation and also in their (unirradiated) F1 progeny. The implications of these findings for the use of the inherited sterility technique for the control of S. litura and other pest Lepidoptera are discussed.

Distribution and Identification of Culex pipiens Linnaeus, 1758 (Diptera: Culicidae) in the Kurdistan Region of Iraq

The specimens utilized for this investigation comprise adult individuals of Culex pipiens and their immature forms. The Culex pipiens mosquitoes are captured using a net trap, while larvae are collected using conventional dipping techniques. The samples are selected at random from fourteen locations in the provinces of the Kurdistan area of Iraq, specifically Erbil, Duhok, Sulaimaniyah and Halabja. From September 15, 2020 to November 20, 2021, a total of 291 female Culex pipiens mosquitoes were collected from the four provinces stated above. The breakdown of the samples is as follows: 135 from one province, 82 from another province, 23 from a third province, and 51 from the fourth province. The diagnosis key is provided for adult individuals and fourth-stage larvae, which originate from a species of mosquito. To illustrate this study, a concise morphological description was utilized, and photos were included to provide a vivid depiction.The findings of the present investigation indicate that adult females had a greater wing length compared to males. The wing length of females ranged from 3.3 to 4 mm, with an average of 3.55 mm. When adult Culex pipiens were looked at, the costa and sub-costal intersections were found at or beyond the point where R2+3 splits in most of the samples. The siphons of fourth-instar larvae are elongated and taper gradually towards the end. On average, their length is 1.4 mm, with a range of 1.3 to 1.5 mm.. KEYWORDS :Culicidae, Culex pipines, Diptera, Kurdistan Region, Iraq, Survey.

The relationship between neurotrophins and cognitive functions in the context of emotional response to sleep deprivation

IntroductionStudies conducted up to date on the subject of deprivation of sleep (DS) primarily focused either on its impact on certain cognitive abilities or mood-enhancing effects in patients with depression. A notable body of evidence suggests that both might be related to alterations in neurotrophin synthesis induced by DS. However, the role of NTs as an interface between DS, mood, and cognitive functions is unclear.ObjectivesThe study aimed to investigate associations between cognitive abilities measured by Trail Making Test (TMT) and Stroop Color and Wort Test (ST), serum protein concentrations of brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), neurotrophin-3 (NT3), neurotrophin-4 (NT4) as well as the expression of their respective genes after a night of sleep deprivation.MethodsEach participant (n=76) underwent a 24-hour DS under the control of actigraphy. Venous blood collection, TMT, and ST were carried out in the morning after DS. Mood was evaluated twice, after DS and in the preceding evening; based on the alleviation of depression symptoms participants were divided into respondents (RE; n=47) and non-respondents (NR; n=29). Serum protein concentration was determined using ELISA kits. Gene expression was evaluated by quantitative real-time polymerase chain reaction with gene-specific probes (reference gene: β-actin). Relative expression was calculated using the Livak formula. TMT is a neuropsychological instrument; Part 1 is thought to evaluate mostly attention, whereas Part 2 executive functions. ST is a 2-part test applied in the assessment of response inhibition and complex attention.ResultsIn RE, cognitive abilities were not associated with expression levels of any of the studied proteins or mRNA (all p&gt;0.05). In NR, BDNF and GDNF mRNA expressions negatively correlated with TMT Part 1 (p=0.017, p=0.048, respectively); scores obtained in TMT Part 2 bore a similar relation to BDNF, GDNF, and NT4 mRNA (p=0.034, p=0.041, p=0.026, respectively). In this group, expression of all BDNF, GDNF, NT3, NT4 mRNA correlated negatively with both parts of ST (p&lt;0.001, p=0.009, p=0.042, p=0.009 for Part 1; p&lt;0.001, p=0.003, p=0.031, p=0.014 for Part 2, respectively).ConclusionsThose results suggest that alterations in the synthesis of NTs might be an element of the molecular milieu characterizing different types of DS response. Negative correlations between test scores and NT mRNA expressions could imply that the reduction of the production of NT proforms might protect against the decline of cognitive functions in the aftermath of DS. Projects using a larger battery of tests as well as analyzing immature forms of NTs would be desirable in order to further investigate mechanisms underlying DS response.Disclosure of InterestNone Declared

One Health Approach on Ehrlichia canis: Serosurvey of Owners and Dogs, Molecular Detection in Ticks, and Associated Risk Factors in Tick-Infested Households of Southern Brazil.

Background: Ehrlichia canis has been the main hemopathogen affecting domestic dogs in Brazil. Even though tick-infested dogs may lead to household infestation and predispose human exposure and public health concern, no comprehensive study has surveyed humans, dogs, and environmental ticks altogether. Materials and Methods: Accordingly, the present study aimed to assess tick-infested households, identify tick species, perform serological (immunofluorescence assay) and molecular (PCR and q-PCR) detection of Ehrlichia in ticks, in the eighth biggest metropolitan area of Brazil. Results: Between 2007 and 2020, 233/5973 (3.9%) out of all complaints were from tick-infested households of 200 different addresses. Overall, 370/552 (67.0%) ticks were collected and identified as adult and 182/552 (33.0%) as immature forms of Rhipicephalus sanguineus s.l. complex; a single tick from one owner, a female tick of Amblyomma sculptum; and 395 ticks from dogs, 319/395 (80.8%) adult and 72/395 (18.2%) immature forms of Rhipicephalus spp., and 4/395 (1.01%) female Amblyomma aureolatum. Overall, 2/135 (1.5%) owners and 13/136 (9.6%) dogs were seropositive for E. canis. The DNA of Anaplasmataceae family was molecularly detected in 16/50 (32.0%) R. sanguineus s.l. As expected, the number of monthly tick infestation complaints were directly associated, and mean (p = 0.01), maximum (p = 0.011), and minimum (p = 0.008) temperature were statistically significant and had a low positive correlation (0.24, 0.23, and 0.24, respectively). In addition, complaints were highly associated to all socioeconomic variables (p < 0.001), with the exception of the presence of vacant lots. Conclusions: Despite low samplings and human negative results, areas with low-income with adequate temperature and urban agglomerations have been shown to be associated risks for tick infestations, predisposing tick-borne diseases. In conclusion, monitoring should always be conducted in such areas, including One Health approach with serosurvey of owners and dogs, along with identification and molecular screening of ticks.

Community-acquired pneumonia in young children living in various climatogeographic zones of the Republic of Dagestan

Introduction. Community-acquired pneumonia (CAP) in early childhood continues to cause significant morbidity and remains one of the most common serious infections in childhood. Each region of the Russian Federation exhibits specific manifestations of CAP morbidity. Knowledge of CAP etiology is crucial for planning therapeutic and preventive measures. &#x0D; Aim. To determine the etiological and immunological features of CAP in early childhood, residing in various climatogeographical zones of the Republic of Dagestan.&#x0D; Materials and methods. The material includes the results of examination and treatment of CAP 204 young children. living in lowland, foothill and mountain climatogeographical zones of the Republic of Dagestan. The selection criterion was the clinical-radiological diagnosis of CAP, using the classification of clinical forms of bronchopulmonary diseases in children.&#x0D; Results. The study revealed a rise in the incidence of respiratory organ diseases, including CAP, among children in the Republic of Dagestan (RD) over the past 5 years. Among infants under one year, the CAP rate was twice as high. The prevalence can be attributed to the climatogeographical conditions of children’s living environments. Streptococcal infection was also observed to predominate in the ethnostructure of CAP in early childhood across all climatogeographical zones. Streptococcus pneumoniae (48.1%) was more frequently isolated in all climatogeographical zones, while Staphylococcus aureus (13.3%) dominated in the mid-mountain zone. In mild pneumonia, cellular immunity components, represented by reduced CD3, CD4, CD25, CD71, react primarily. Disease exacerbation leads to depletion of CD8, CD16, CD20. Severe pneumonia triggers the involvement of humoral immunity, leading to changes in clinical blood analysis — leukocytosis and an increase in mature and immature forms of neutrophils. The levels of IL-1, IL-6, IL-8 in blood serum increase depending on the area of inflammation and the CAP severity. In observed children with focal-draining and segmental CAP, their levels significantly increased. For empirical antibacterial therapy of CAP, recommendations include amoxicillin, protected ampicillins, third-generation cephalosporins, macrolides, and vancomycin in mono- or combination therapy.&#x0D; Conclusion. The conducted research highlights the need for managing ethnological monitoring with antibiotic sensitivity testing of infectious agents causing CAP in children.

Unraveling Dengue Dynamics: In-Depth Epidemiological and Entomological Analyses in Bengaluru, India.

In view of the increased number of detected dengue cases in Bengaluru, a request for situation analysis was received from local health authorities in the selected area. The study included epidemiological and entomological assessments to understand the same. The immature forms collected were allowed to emerge, pooled, and processed for vector incrimination. In the surveyed population (347), 20 (5.8%) reported fever cases and 12 (3.5%) were confirmed as dengue cases among the 102 families. Stegomyia indices were high. Vector incrimination studies revealed pools positive for the presence of dengue virus in flower pots, fridge trays, plastic barrels, and rubber tires habitats. This study highlights the increased risk of dengue fever incidence in communities wherepoor intra and peri-domestic sanitation practices are prevailing and recommendsregular entomological surveillance of denguevirus in its vector population..

Symptomatic post COVID patients have impaired alveolar capillary membrane function and high VE/VCO2

BackgroundPost COVID-19 syndrome is characterized by several cardiorespiratory symptoms but the origin of patients’ reported symptomatology is still unclear.MethodsConsecutive post COVID-19 patients were included. Patients underwent full clinical evaluation, symptoms dedicated questionnaires, blood tests, echocardiography, thoracic computer tomography (CT), spirometry including alveolar capillary membrane diffusion (DM) and capillary volume (Vcap) assessment by combined carbon dioxide and nitric oxide lung diffusion (DLCO/DLNO) and cardiopulmonary exercise test. We measured surfactant derive protein B (immature form) as blood marker of alveolar cell function.ResultsWe evaluated 204 consecutive post COVID-19 patients (56.5 ± 14.5 years, 89 females) 171 ± 85 days after the end of acute COVID-19 infection. We measured: forced expiratory volume (FEV1) 99 ± 17%pred, FVC 99 ± 17%pred, DLCO 82 ± 19%, DM 47.6 ± 14.8 mL/min/mmHg, Vcap 59 ± 17 mL, residual parenchymal damage at CT 7.2 ± 3.2% of lung tissue, peakVO2 84 ± 18%pred, VE/VCO2 slope 112 [102–123]%pred. Major reported symptoms were: dyspnea 45% of cases, tiredness 60% and fatigability 77%. Low FEV1, Vcap and high VE/VCO2 slope were associated with persistence of dyspnea. Tiredness was associated with high VE/VCO2 slope and low PeakVO2 and FEV1 while fatigability with high VE/VCO2 slope. SPB was fivefold higher in post COVID-19 than in normal subjects, but not associated to any of the referred symptoms. SPB was negatively associated to Vcap.ConclusionsIn patients with post COVID-19, cardiorespiratory symptoms are linked to VE/VCO2 slope. In these patients the alveolar cells are dysregulated as shown by the very high SPB. The Vcap is low likely due to post COVID-19 pulmonary endothelial/vasculature damage but DLCO is only minimally impaired being DM preserved.

Molecular identification of immature Cerambycinae (Coleoptera: Cerambycidae): revisiting the significance of immature morphologies

Abstract The microstructure of immature insect forms plays a crucial role in systematics, especially in inferring phylogenetic relationships and designating the morphological basis for taxonomic units. However, the microstructure of longhorn beetle larvae and pupae is poorly understood, owing to difficulties in obtaining them and technical limitations. Among 90 larvae and 80 pupae collected and photographed, 31 larvae and 15 pupae remained unidentified after morphological and bionomical evidence-based identification. Cytochrome c oxidase I sequences were amplified successfully from 68 larvae and 59 pupae. Molecular identification using tree-based analysis identified 28 of 31 undetermined larvae (90.3%) and 14 of 15 undetermined pupae (93.3%) to the species level. We highlight the usefulness of DNA barcoding in identifying immature forms of cerambycid beetles and filling gaps in public databases. This study also demonstrates how microstructure can clarify the morphological basis of polyphyletic taxa on the phylogeny and emphasizes the significance of the microstructure of immature stages in longhorn beetle systematics.

Immature Sacrococcygeal Teratoma: A Case Report and Extensive Review of the Literature

Immature sacrococcygeal teratoma represents a histological form with rapid tumor growth, a risk of premature birth, an enhanced rate of complications, an increased risk of recurrence, and a higher mortality rate than the mature type. Thus, prenatal diagnosis of immature forms would significantly improve the prognosis of these cases. To this end, we performed an extensive literature review on the diagnosis, therapeutic management, and follow-up of immature teratomas. Regarding this medical conduct, we also presented our case. In conclusion, the early identification of immature sacrococcygeal teratomas with or without other associated structural abnormalities and their correct therapeutic approach are basic principles for a favorable evolution of these cases.

Influence of pH on protein extraction from Sus scrofa pancreas

The porcine pancreas contains various enzymes, structural, regulatory, secretory, receptor and other biologically active substances that ensure both the functioning of the organ and its biological role in the organism. The aim of this work was to study the influence of pH changes in 0.9% sodium chloride solution used as an extractant on the efficiency of bioactive protein isolation from the porcine pancreas. The extraction was carried out with the 0.9% NaCl, 0.9% NaCl pH=4 and 0.9% NaCl pH= 8.5 with a stirring speed of 400 rpm for 150 min at 4 ºC; the ratio of pancreas: extractant was 1:5, the supernatant was separated by centrifugation. The protein concentration was measured by a biuret reaction on a semi-automatic biochemical analyzer Biochem SA. The proteomic composition of extracts and native pancreas was evaluated by 10% SDS-PAGE according to Laemmli method in the “VE10” chamber. Digital images of electrophoregrams were obtained using a Bio-5000 Plus scanner, edited in a graphic editor and analyzed using ImageJ software. When determining the intensity of protein fractions, it was noted that the use of 0.9% NaCl contributed to a greater yield of proteins with molecular weights of 200 kDa, 150 kDa, 69 kDa, 52 kDa and 33 kDa into the extractant; a pH shift to the acidic area stimulated the yield of fractions with molecular weights of 130 kDa, 50 kDa, 49 kDa, 45 kDa, 40 kDa, 30 kDa and 27kDa, and a pH shift to the alkaline area — only 47 kDa and 42 kDa. Most pancreas proteolytic enzymes have a molecular weight in a range of 34–23kDa, excepting the immature form of carboxypeptidases with MW 45–47kDa. The greatest intensity of protein bands was observed in the region with MW less than 33kDa on the obtained electrophoregrams. The presence of intense protein fractions in the region of molecular weights of less than 50–52kDa and 40kDa was also noted, which may correspond to enzymes such as pancreatic lipase and phospholipase A2, and the presence of protein fractions with MW above 130 kDa corresponding to various types and isoforms of collagen and laminin. In addition, such processes as protein aggregation and proteolysis can also influence the molecular weight distribution of protein fractions.