Microbial diversity in selected Indonesian marine organisms

Metagenomics, the study of genetic material taken directly from the environment, generally includes the analysis of a whole microbial community, from both a taxonomic and functional perspective. In contrast to traditional microbiological techniques, this approach makes the isolation and cultivation of single organisms unnecessary. This allows the investigation of non-cultivable microorganisms, which represent the majority of microorganisms. While most metagenomic studies have focused on human and model organisms, the metagenome of marine species is relatively unknown. One area in particular that could be of major interest for aquaculture, but which has not been sufficiently investigated, is the influence of the environment on the composition of the bacterial community. This work deals with the effects that highly polluted and non-polluted environmental conditions have on bacterial communities. To do this, the microbial communities of three fish species (Atule mate, Epinephelus sexfasciatus and Epinephelus fuscoguttatus) and one shrimp species (Penaeus monodon) were investigated. Two different metagenomic techniques were applied. The results of a 16S amplicon-based approach did not reveal any differences between the investigated environments at a high taxonomic level. However, differences between the sampling locations could be found. A subsequent whole metagenome sequencing approach revealed, that predominant members of the microbiome living within a controlled environment of a mariculture facility are more stably distributed than those within free-living host species. Furthermore, the interactions between endoparasites and potential pathogenic bacteria were investigated. An analysis of the microbiome of shrimp shows only minor differences in the microbial community composition between free-living and aquacultured Penaeus monodon. In contrast, the microbial communities of the polluted environment showed significant differences

Triple platelet inhibition in intracranial thrombectomy with additional acute cervical stent angioplasty due to tandem lesion: a retrospective single-center analysis.

BACKGROUND Acute stroke treatment with intracranial thrombectomy and treatment of ipsilateral carotid artery stenosis/occlusion ("tandem lesion", TL) in one session is considered safe. However, the risk of stent restenosis after TL treatment is high, and antiplatelet therapy (APT) preventing restenosis must be well balanced to avoid intracranial hemorrhage. We investigated the safety and 90-day outcome of patients receiving TL treatment under triple-APT, focused on stent-patency and possible disadvantageous comorbidities. METHODS Patients receiving TL treatment in the setting of acute stroke between 2013 and 2022 were analyzed regarding peri-/postprocedural safety and stent patency after 90 days. All patients received intravenous eptifibatide and acetylsalicylic acid and one of the three drugs prasugrel, clopidogrel, or ticagrelor. Duplex imaging was performed 24 h after treatment, at discharge and 90 days, and digital subtraction angiography was performed if restenosis was suspected. RESULTS 176 patients were included. Periprocedural complications occurred in 2.3% of the patients at no periprocedural death, and in-hospital death in 13.6%. Discharge mRS score was maintained or improved at the 90-day follow-up in 86%, 4.54% had an in-stent restenosis requiring treatment at 90 days. No recorded comorbidity considered disadvantageous for stent patency showed statistical significance, the duration of the endovascular procedure had no significant effect on outcome. CONCLUSION In our data, TL treatment with triple APT resulted in a low restenosis rate, low rates of sICH and a comparably high number of patients with favorable outcome. Aggressive APT in the initial phase may therefore have the potential to prevent recurrent stroke better than restrained platelet inhibition. Comorbidities did not influence stent patency

Drug-Coated Balloons for Treatment of Internal Carotid Artery Restenosis After Stenting: A Single-Center Mid-Term Outcome Study.

PURPOSE Endovascular and surgical treatments of stenosis of the extracranial internal carotid artery (ICA) are common procedures, yet both introduce a risk of restenosis due to endothelial hyperplasia. Drug-coated balloons (DCBs) are designed to decrease neointimal hyperplasia, however rarely used in the neurovascular setting. This study retrospectively analyzes mid-term results of DCB-treated in-stent restenosis (ISR) of the ICA. MATERIALS AND METHODS The medical history, comorbidities, and periprocedural data of patients receiving DCB treatment for > 50% ISR of the ICA after carotid artery stenting were analyzed. Follow-up after DCB treatment was performed with Doppler ultrasound. Suspicious cases were checked with CT- or MR-angiography and-if there was agreement between the modalities-validated with digital subtraction angiography. Potential risk factors for restenosis and differences in outcomes after PTA with three types of DCB balloons were evaluated. RESULTS DCB treatment was performed in 109 cases, 0.9% of which involved in-hospital major stroke; no minor strokes occurred. A total of 17 patients (15.6%) had recurrent ISR after DCB treatment, after a mean time of 30.2 months (7-85 months). Tobacco use was significantly associated with a higher incidence of recurrent ISR. CONCLUSION DCB angioplasty for ISR is an effective treatment that may delay and decrease restenosis. Treating comorbidities and adopting lifestyle changes may additionally help prevent ISR

Metagenomic analysis between free-living and cultured Epinephelus fuscoguttatus under different environmental conditions in Indonesian waters

Abstract not availablePhilipp Hennersdorf, Grit Mrotzek, Muslihudeen A. Abdul-Aziz, Hans Peter Salu

Spectrum of the pH at given time points (6 h and 10 h p.i.) in apoptotic and non-apoptotic infected monocytes.

<p>The dashed lines (shown in blue and green) are the spectra of pH in wild-type (wt.) and mutant infection in non-apoptotic monocyte at 6 h p.i. when their pH showed approximately similar values (6.3 to 6.5). The constant lines represent the wild-type or mutant infections in apoptotic monocyte (treated with STS) comparing two time points of 6 and 10 h p.i. The most acidic pH (less than 4) was recorded 10 h p.i. in the apoptotic monocyte infected with mutant whereas at the same time point the apoptotic monocyte infected with wild-type showed a higher pH around 5.3.Regardless of the type of infection, the signals related to acidic pH in non-apoptotic cells were generally less intense than in apoptotic cells. 10 h p.i., the apoptotic cells infected by melanin-free <i>pksP</i> mutant conidia were far more acidic compared to cells containing wild-type conidia.</p

Hyperspectral Imaging Using Intracellular Spies: Quantitative Real-Time Measurement of Intracellular Parameters <i>In Vivo</i> during Interaction of the Pathogenic Fungus <i>Aspergillus fumigatus</i> with Human Monocytes

<div><p>Hyperspectral imaging (HSI) is a technique based on the combination of classical spectroscopy and conventional digital image processing. It is also well suited for the biological assays and quantitative real-time analysis since it provides spectral and spatial data of samples. The method grants detailed information about a sample by recording the entire spectrum in each pixel of the whole image. We applied HSI to quantify the constituent pH variation in a single infected apoptotic monocyte as a model system. Previously, we showed that the human-pathogenic fungus <i>Aspergillus fumigatus</i> conidia interfere with the acidification of phagolysosomes. Here, we extended this finding to monocytes and gained a more detailed analysis of this process. Our data indicate that melanised <i>A</i>. <i>fumigatus</i> conidia have the ability to interfere with apoptosis in human monocytes as they enable the apoptotic cell to recover from mitochondrial acidification and to continue with the cell cycle. We also showed that this ability of <i>A</i>. <i>fumigatus</i> is dependent on the presence of melanin, since a non-pigmented mutant did not stop the progression of apoptosis and consequently, the cell did not recover from the acidic pH. By conducting the current research based on the HSI, we could measure the intracellular pH in an apoptotic infected human monocyte and show the pattern of pH variation during 35 h of measurements. As a conclusion, we showed the importance of melanin for determining the fate of intracellular pH in a single apoptotic cell.</p></div

Microbial Diversity and Parasitic Load in Tropical Fish of Different Environmental Conditions.

In this study we analysed fecal bacterial communities and parasites of three important Indonesian fish species, Epinephelus fuscoguttatus, Epinephelus sexfasciatus and Atule mate. We then compared the biodiversity of bacterial communities and parasites of these three fish species collected in highly polluted Jakarta Bay with those collected in less polluted Indonesian areas of Cilacap (E. sexfasciatus, A. mate) and Thousand Islands (E. fuscoguttatus). In addition, E. fuscoguttatus from net cages in an open water mariculture facility was compared with free living E. fuscoguttatus from its surroundings. Both core and shared microbiomes were investigated. Our results reveal that, while the core microbiomes of all three fish species were composed of fairly the same classes of bacteria, the proportions of these bacterial classes strongly varied. The microbial composition of phylogenetically distant fish species, i.e. A. mate and E. sexfasciatus from Jakarta Bay and Cilacap were more closely related than the microbial composition of more phylogentically closer species, i.e. E. fuscoguttatus, E. sexfasciatus from Jakarta Bay, Cilacap and Thousand Islands. In addition, we detected a weak negative correlation between the load of selected bacterial pathogens, i.e. Vibrio sp. and Photobacterium sp. and the number of endoparasites. In the case of Flavobacterium sp. the opposite was observed, i.e. a weak positive correlation. Of the three recorded pathogenic bacterial genera, Vibrio sp. was commonly found in E. fuscoguttatus from mariculture, and lessly in the vicinity of the net cages and rarely in the fishes from the heavily polluted waters from Jakarta Bay. Flavobacterium sp. showed higher counts in mariculture fish and Photobacteria sp. was the most prominent in fish inside and close to the net cages

Effects of chloroquine treatment and phagosomal acidification on <i>A</i>.<i>fumigatus</i> infection in an apoptotic cell.

<p>The acidified pH of the apoptotic cell in the presence of chloroquine was maintained after infection. As it can be concluded, <i>A</i>.<i>fumigatus</i> only interferes with pH if the phagolysosme stays intact.</p

Spectrum of the pH at given time points (6 h and 10 h p.i.) in apoptotic and non-apoptotic infected monocytes.

<p>The dashed lines (shown in blue and green) are the spectra of pH in wild-type (wt.) and mutant infection in non-apoptotic monocyte at 6 h p.i. when their pH showed approximately similar values (6.3 to 6.5). The constant lines represent the wild-type or mutant infections in apoptotic monocyte (treated with STS) comparing two time points of 6 and 10 h p.i. The most acidic pH (less than 4) was recorded 10 h p.i. in the apoptotic monocyte infected with mutant whereas at the same time point the apoptotic monocyte infected with wild-type showed a higher pH around 5.3.Regardless of the type of infection, the signals related to acidic pH in non-apoptotic cells were generally less intense than in apoptotic cells. 10 h p.i., the apoptotic cells infected by melanin-free <i>pksP</i> mutant conidia were far more acidic compared to cells containing wild-type conidia.</p

Kinetics of phagolysosomal pH upon infection.

<p><b>(A)</b> Phagosomal pH in an apoptotic monocyte containing labelled wild-type conidia in comparison to an apoptotic monocyte carrying <i>pksP</i> mutant conidia. <b>(B)</b> Data represent the mean ₊ SD from three experiments. <b>(C)</b> Phagosomal pH in the survived apoptotic monocyte infected with labelled wild-type conidia. <b>(D)</b> Data represent the mean ₊ SD of cytosolic pH from three experiments.</p