Uranium mining effluents: What about the re-use of mining wastes to improve the bioproduction of industrially relevant bioactive compounds?

The shift towards a circular economy, where waste generation is minimized through waste re-use and the development of valorization strategies, is crucial for the establishment of a low carbon, sustainable, and resource-efficient economy. However, there is a lack of strategies for re-using and valorizing specific types of waste, particularly those containing naturally occurring radioactive materials (NORM), despite the prevalence of industrial activities that produce such waste due to their chemical and radiological hazards. Living organisms, including fungi, are valuable sources of bioactive compounds with various industrial applications. In this study, we assessed the growth and metabolic profile changes of three white rot fungi species in response to low concentrations of a uranium mine effluent containing NORM and metals to explore their potential for producing biotechnologically relevant bioactive compounds. The growth rate was assessed in three different culture media, with and without the uranium mine effluent (1% V/V)), and the metabolic profile was analyzed using FTIR-ATR spectroscopy. Results suggested an improvement in growth rates in media containing the uranium mine effluent, although not statistically significant. T. versicolor showed promise in terms of bioactive compound production. The production of droplets during growth experiments and significant metabolic changes, associated with the production of bioactive compounds like laccase, melanin, and oxalic acid, were observed in T. versicolor grown in mYEPDA with the uranium mine effluent. These findings present new research opportunities for utilizing waste to enhance the biotechnological production of industrially relevant bioactive compounds and promote the development of circular economy strategies for re-using and valorizing NORM-containing waste.

Ultrasound-Guided WALANT Technique in Carpal Tunnel Decompression Surgery.

Carpal tunnel syndrome (CTS) is the most common compressive neuropathy in the human body. Its symptoms result from compression of the median nerve in the carpus. The treatment can be conservative, with medications and/or infiltrations that alleviate the symptoms, or surgical, which is more effective, with decompression of the median nerve by surgical section of the flexor retinaculum of the carpus. The anesthetic technique varies according to the anesthesia method: sedation, venous locoregional anesthesia and, more recently, wide-awake local anesthesia no tourniquet (WALANT), which can be performed by the surgeons themselves. The WALANT technique uses local anesthesia with a vasoconstrictor, and does not require the use of a tourniquet on the upper limb nor sedation. The median nerve block in ultrasound-guided WALANT provides better accuracy to the technique, with greater patient safety; in the present article, its use in the performance of carpal tunnel decompression is described, and the literature is reviewed.

Ultrasound-Guided WALANT Technique in Carpal Tunnel Decompression Surgery* / Técnica WALANT guiada por ultrassom na cirurgia de descompressão do túnel do carpo

Abstract Carpal tunnel syndrome (CTS) is the most common compressive neuropathy in the human body. Its symptoms result from compression of the median nerve in the carpus. The treatment can be conservative, with medications and/or infiltrations that alleviate the symptoms, or surgical, which is more effective, with decompression of the median nerve by surgical section of the flexor retinaculum of the carpus. The anesthetic technique varies according to the anesthesia method: sedation, venous locoregional anesthesia and, more recently, wide-awake local anesthesia no tourniquet (WALANT), which can be performed by the surgeons themselves. The WALANT technique uses local anesthesia with a vasoconstrictor, and does not require the use of a tourniquet on the upper limb nor sedation. The median nerve block in ultrasound-guided WALANT provides better accuracy to the technique, with greater patient safety; in the present article, its use in the performance of carpal tunnel decompression is described, and the literature is reviewed.

Resumo A síndrome do túnel do carpo (STC) é a neuropatía compressiva mais comum do corpo humano. Seus sintomas decorrem da compressão do nervo mediano no carpo. O tratamento pode ser incruento, com medicações e/ou infiltrações que amenizam os sintomas, ou cruento, mais eficaz, com a descompressão do nervo mediano pela seção cirúrgicadoretináculodos flexores do carpo. A técnica anestésica varia de acordo com o serviço de anestesia: sedação, anestesia locorregional venosa e, mais recentemente, a anestesia local com o paciente acordado e sem torniquete (wide-awake local anesthesia no tourniquet, WALANT), que pode ser realizada pelo próprio cirurgião. Por utilizar anestesia local com vasoconstritor, essa técnica dispensa o uso de torniquete no membro superior e a necessidade de sedação. O bloqueio do nervo mediano na WALANT guiada por ultrassonografiaconfere melhor precisão àtécnica,e mais segurança ao paciente, e, neste artigo seu uso na realização da descompressão do túnel do carpo é descrito, e a literatura, revisada.

Comparative analyses of Mikania (Asteraceae: Eupatorieae) plastomes and impact of data partitioning and inference methods on phylogenetic relationships.

We assembled new plastomes of 19 species of Mikania and of Ageratina fastigiata, Litothamnus nitidus, and Stevia collina, all belonging to tribe Eupatorieae (Asteraceae). We analyzed the structure and content of the assembled plastomes and used the newly generated sequences to infer phylogenetic relationships and study the effects of different data partitions and inference methods on the topologies. Most phylogenetic studies with plastomes ignore that processes like recombination and biparental inheritance can occur in this organelle, using the whole genome as a single locus. Our study sought to compare this approach with multispecies coalescent methods that assume that different parts of the genome evolve at different rates. We found that the overall gene content, structure, and orientation are very conserved in all plastomes of the studied species. As observed in other Asteraceae, the 22 plastomes assembled here contain two nested inversions in the LSC region. The plastomes show similar length and the same gene content. The two most variable regions within Mikania are rpl32-ndhF and rpl16-rps3, while the three genes with the highest percentage of variable sites are ycf1, rpoA, and psbT. We generated six phylogenetic trees using concatenated maximum likelihood and multispecies coalescent methods and three data partitions: coding and non-coding sequences and both combined. All trees strongly support that the sampled Mikania species form a monophyletic group, which is further subdivided into three clades. The internal relationships within each clade are sensitive to the data partitioning and inference methods employed. The trees resulting from concatenated analysis are more similar among each other than to the correspondent tree generated with the same data partition but a different method. The multispecies coalescent analysis indicate a high level of incongruence between species and gene trees. The lack of resolution and congruence among trees can be explained by the sparse sampling (~ 0.45% of the currently accepted species) and by the low number of informative characters present in the sequences. Our study sheds light into the impact of data partitioning and methods over phylogenetic resolution and brings relevant information for the study of Mikania diversity and evolution, as well as for the Asteraceae family as a whole.

Metals and low dose IR: Molecular effects of combined exposures using HepG2 cells as a biological model.

Uranium mining sites produce residues rich in metals and radionuclides, that may contaminate all environmental matrices, exposing human and non-human biota to low doses of ionizing radiation (LDIR) and to the chemical toxicity of several metals. To date, experimental and radio-epidemiological studies do not provide conclusive evidence of LDIR induced cancer. However, co-exposures (LDIR plus other contaminants), may increase the risks. To determine the potential for genotoxic effects in human cells induced by the exposure to LDIR plus metals, HEPG2 cells were exposed to different concentrations of a uranium mine effluent for 96 h. DNA damage was evaluated using the comet assay and changes in the expression of tumor suppressor and oncogenes were determined using qPCR. Results show that effluent concentrations higher than 5%, induce significant DNA damage. Also, a significant under-expression of ATM and TP53 genes and a significant overexpression of GADD45a gene was observed. Results show that the exposure to complex mixtures cannot be disregarded, as effects were detected at very low doses. This study highlights the need for further studies to clarify the risks of exposure to LDIR along with other stressors, to fully review the IR exposure risk limits established for human and non-human biota.

Shiga toxin-producing Escherichia coli in wild ungulates.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic pathogens that live in the gastrointestinal tract of wildlife and cattle without causing disease. In humans, their colonization and infection lead to life-threatening disease. We investigated the occurrence of STEC in wild ungulates (wild boar, red deer and roe deer) inhabiting areas differently impacted by anthropogenic activities. STEC were detected in 9% (n = 6) of the samples and were recovered from the three species: 1 of wild boar, 4 of red deer and 1 of roe deer. All the isolates (n = 7) were non-O157 STEC encoding stx1 (n = 2; 29%) and/or stx2 genes (n = 6; 86%). O27:H30 was the most frequent serotype identified, followed by O146:H21 and O146:H28. Two STEC were O-untypable: ONT:H28 and ONT:H52. The phylo-groups identified were B1 (n = 3), E (n = 3) and F (n = 1). All the isolates recovered were susceptible to the different classes of antibiotics tested, although resistance genes were found in two strains. Apart from stx, all STEC encode many virulence factors (VF), particularly adhesins and/or other toxins. A strain with 13 VF collected from roe deer has a high enterohemorrhagic risk due to the presence of intimin, hemolysin and protease effectors genes. Enterohemorrhagic E. coli (EHEC) are implicated in the major cases of human infection and mortality, highlighting the zoonotic potential of wildlife-associated STEC. Wild ungulates are reservoirs of STEC potentially pathogenic to humans. Therefore, following the One Health concept, it is crucial to establish worldwide local monitoring programs that will benefit human, animal and environmental health.

The lanthipeptides of Bacillus methylotrophicus and their association with genomic islands.

Bacillus methylotrophicus strains are known for their potential as plant-growth promoters and as microbial pesticides that effectively control plant diseases caused by bacteria and fungi. Over the past few years, a wide diversity of their secondary metabolites has been extensively characterized. Among these are the RiPPs lanthipeptides, which are an important and growing group of notable compounds. The increasing interest in B. methylotrophicus species, accompanied by the development of high throughput sequencing techniques, has resulted in a substantial number of full genomes being available. Here, an in silico analysis was performed on these genomes in order to survey the presence of lanthipeptide biosynthetic clusters. It was found that the pan genome of B. methylotrophicus only encoded the biosynthesis of mersacidin and amylolysin, which are lanthipeptides with antibacterial activity. However, the amylolysin gene cluster identified was comprised of more genetic elements than those previously described, and it had certain features of two-peptide lantibiotics. Additionally, it was also established that the association of lanthipeptides with genomic islands (GIs) was not confined to mersacidin. This was also found for the amylolysin cluster as well as other class I and class II lanthipeptides, supporting the idea that their production is probably related to functional adaptation.

Bioengineering of lanthipeptides in Escherichia coli: assessing the specificity of lichenicidin and haloduracin biosynthetic machinery.

The lichenicidin and haloduracin biosynthetic machinery specificity was investigated in vivo in Escherichia coli. Unlike previous reports using different hosts, it was found that the biosynthetic machineries of lichenicidin and haloduracin are highly specific to their dedicated peptide precursors. Likewise, the substitution of lichenicidin structural genes by chimeras of lichenicidin leader sequences and haloduracin core peptides did not yield mature haloduracin peptides. Despite these restrictions, it was found that the bifunctional enzyme HalT was able to process and export lichenicidin peptides. These findings corroborate the promiscuity of LanT enzymes reported for other lantibiotics, such as nukacin ISK-1 and lacticin 481.

Biomonitoring a human population inhabiting nearby a deactivated uranium mine.

Environmental exposure to uranium and its daughter radionuclides, has been linked to several negative effects such as those related with important physiological processes, like hematopoiesis, and may also be associated with genotoxicity effects. Herein, genotoxic effects, immunotoxicity, trace elements and C reactive protein (CRP) analyses, were performed in peripheral blood samples collected from individuals of a population living near a deactivated uranium mine. C reactive protein analysis was performed to exclude candidates with active inflammatory processes from further evaluations. DNA damage and immunotoxicity (immunophenotyping and immune cell counts) were evaluated by comet assay and flow cytometry, respectively. Significant DNA damage was observed in the peripheral blood samples from volunteers living in the Cunha Baixa village. A significant decrease of NK and T lymphocytes counts were observed in the individuals from the Cunha Baixa village, when compared with individuals from the reference site. Uranium and manganese levels were significantly higher in the Cunha Baixa village inhabitants. On the other hand, zinc levels were significantly lower in those individuals when compared with the volunteers from the control village. Results suggest that inhabitants from Cunha Baixa have a higher risk of suffering from serious diseases such as cancer, since high DNA damages were observed in peripheral blood leukocytes and also decreased levels of NK and T cells, which play an essential role in the defense against tumor growth.

A novel complex class 1 integron found in a Klebsiella pneumoniae isolate from Portugal.

Klebsiella pneumoniae Kp1 carrying a novel complex class 1 integron was isolated from an inanimate surface of a female ward sanitary facility in the Hospital Infante D. Pedro, Aveiro, central Portugal. The integron consists of two variable regions (VRs); VR1 was previously described in Escherichia coli and Vibrio cholerae, and VR2 contains an In37-like structure and is located downstream of an ISCR1 element. The integron was found on a plasmid of 225 kb. The qnrB10 gene, although present, is not associated with the complex class 1 integron.

Tn5090-like class 1 integron carrying bla(VIM-2) in a Pseudomonas putida strain from Portugal.

Three Pseudomonas putida strains containing bla(VIM-2) were isolated from an inanimate surface of a female ward sanitary facility in the Hospital Infante D. Pedro, Aveiro. A novel class 1 integron was found in strain Pp2 (aacA4/bla(VIM-2)/aac6'-IIc disrupted by an insertion sequence IS1382), and strain Pp1 was found to carry a class 1 integron (aacA7/bla(VIM-2)/aacC1/aacA4), which is described for the first time in this species. Strain PF1 carries a class 1 integron associated with a Tn5090-like transposon, constituting the first finding of this type of arrangement in a strain from Portugal. This association highlights further dissemination of bla(VIM-2) in environmental hospital isolates.

In99, an In100-related integron, its occurrence and prevalence in clinical Pseudomonas aeruginosa strains from a central region of Portugal.

In99, a possible ancestor of In100, is a class 1 integron associated with carbenicillinase (blaPSE) and aminoglycoside resistance genes [aac(6')-Ib and aadA2]. In99 was present in 8 of 81 clinical isolates of Pseudomonas aeruginosa from unrelated patients collected in different years. The strains fell into two clonal groups and exhibited resistance to beta-lactams and aminoglycosides.