Dust samples were collected from air vents and ground surfaces, and the total DNA was analyzed for bacteria and fungi by amplifying 16S rRNA and ITS genes on the Illumina Miseq. Results indicated that Leptosphaerulina was the most abundant fungal genus present in the air vent and ground samples from HNM and HVM. At the same time, the most abundant bacterial genera in the air vent and ground samples were Propionibacterium and Streptococcus. The fungi community diversity was significantly different in the air vent samples. The abundance of fungal species known to be associated with respiratory diseases in indoor dust samples was similar, regardless of the visibility of fungi in the houses. The existence of fungi associated with respiratory symptoms was compared with several parameters like dust particulate matter (PM), CO2 level, temperature, and humidity. Most of these parameters are either positively or negatively correlated with the existence of fungi associated with respiratory diseases; however, none of these correlations were significant at p = 0.05. Our results indicate that implementing molecular methods for detecting indoor fungi may strengthen common exposure and risk assessment practices.The presence of fungi in the indoor environment is associated with allergies and other respiratory symptoms. The aim of this study was to use sequencing and molecular methods, including next-generation sequencing (NGS) approaches, to explore the bacterial and fungal communities and their abundance in the indoor environment of houses ( The indoor environment is believed to have a significant health impact as people spend the majority of their time in a built environment. Past research has gone so far as to estimate that humans in industrialized countries spend up to 90% of their lives inside ,2. SeverPenicillium, Aspergillus, and Cladosporium, which may be associated with breathing problems [Penicillium sp., associated with asthma, and Aspergillus sp., associated with atopy, are the primary sources of fungal loads in these indoor spaces due to their ability to grow at high concentrations on substrates such as wallpaper or upholstery [Aspergillus fumigatus (AF) [Emerging evidence suggests that damp houses favor the growth of fungal spores formed by problems ,10. The problems . Fortunaproblems . Howeverholstery ,14,15,16tus (AF) ,18,19. Atus (AF) . In additus (AF) .In the past, microbial analyses were predominately conducted by culturing. However, the vast majority (estimated at 99% or more) of microorganisms cannot be cultured . It is aMolecular culture-independent approaches, such as quantitative polymerase chain reaction (qPCR), can assess the abundance and bypass the bias of morphological similarities in fungal communities . Howeverhomes with visible mold (HVM) contamination and homes with no visible mold (HNM) in Memphis, TN, USA. Memphis is located near the running edge of the Mississippi River with humidity levels that often range between the 1950s and mid-1980s, which is 20% higher than the EPAs standards to reduce fungal presence [In this study, we used NGS along with qRT-PCR to investigate the indoor microbiome, explicitly focusing on the fungal and bacterial communities of presence , which mThis study was performed in greater Memphis, TN, USA. Samples (described below) were collected using two recruitment methods. The first recruitment utilized the citizens’ mold remediation hotline of the Shelby County Health Department (SCHD). The second method utilized friends’ participation. The friend recruitment was based on known citizens in Memphis who requested participation because of fungal concerns and general air quality in their homes. These subjects did not have any visible fungi before or during the sampling period and therefore were categorized as the control group.2 logger placed at a central location in the living room. During the home setup, the field sheet logged the date, time, location, and other important parameters. The questionnaire asked about relevant information used to gather specific data that could help the project’s objectives and aims. Samples were collected using an ORECK vacuum and respected filter attachment (DUSTREAM Collector) that captured the vacuumed dust content following the Vacuum Dust Sample Collection Protocol recommended by the U.S. Department of Housing and Urban Development (HUD) [A total of twenty houses were sampled during the study. Ten of the twenty houses had visible fungi , and ten houses did not . Field sampling was a one-week test in the respective homes. Samples were taken from the home by vacuuming content, deploying environmental instruments/samplers, and filling out a field sheet and questionnaire. The instruments included a PM counter and a COnt (HUD)  with min2), humidity, and temperature in all the sampled houses (both HNM and HVM). We have used a continuous PM counter  that could measure mass concentrations of small particles. Similarly, the CO2 datalogger  could continuously measure the level of CO2 (ppm), humidity (%), and temperature (°C) using Gaslab software. The PM counter is already deployable after removing any excel sheets from its memory bank; no calibrations are needed for the PM counter as the software is built into the machine, whereas CO2 dataloggers were configured by clearing all previous logs, properly setting the time/date, and adjusting the collection interval to every 5 min.The microbial community largely depends on different types of environmental factors, and thus we have measured dust particulate matter (PM), carbon dioxide (COn = 40) using the Qiagen DNA PowerSoil kit, following the manufacturer’s protocol. Purification of the extracted DNA was performed using the Epigentek DNA Concentrator kit (P-1006). The V4 hypervariable region of the 16S rRNA of the metagenomic DNA was amplified using a set of universal primers 515F (5′-GTGYCAGCMGCCGCGGTAA-3′) and 806R (5′-GGACTACNVGGGTWTCTAAT-3′). Similarly, ITS1F (5′-CTTGGTCATTTAGAGGAAGTAA-3′) and ITS2R (5′-GCTGCGTTCTTCATCGATGC-3′) were used to amplify the ITS region of fungi. The forward primer was tagged with a barcode sequence. The PCR (28 cycles) was conducted with the HotStarTaq Plus Master Mix Kit , following the manufacturer’s instructions. The confirmation of the amplification was conducted by checking the PCR product on a 2% agarose gel. In order to prepare the Illumina DNA library, an equal proportion of the PCR products (in terms of concentration and molecular weight) were pooled and purified by Ampure XP beads. The paired-end sequencing was conducted at MR DNA  on a MiSeq, following the manufacturer’s guidelines.Genomic DNA (gDNA) was extracted from each sample  , with soFungal species targets were selected based on their inclusion in a panel called the Environmental Relative Moldiness Index (ERMI) –developed by the EPA- to evaluate mold issues in water-damaged homes. Most of the species selected in this study are from ERMI Group 1 and Group 2, known to be associated with allergic respiratory diseases, nosocomial infections, and allergic respiratory reactions ,40,41. Thttp://bioinforma tics.psb.ugent.be/webtools/Venn/, accessed on 12 April 2023). All heatmaps are generated using TBtools software [The statistical analysis was conducted using R . R packages ‘Vegan’  were usesoftware .Propionibacterium, Sphingomonas, Staphylococcus, Corynebacterium, Methylobacterium, and Halospirulina . The percentage abundance of bacterial and fungal genera in the air vent and ground samples of both HNM and HVM are shown in pirulina A,B. Howeacterium C,D. Simiergillus A,B. On tbasidium C,D.p < 0.05. Similarly, the diversity of fungal communities in HNM and HVM was calculated and presented in p < 0.05 level.The bacterial and fungal diversity in ground and air vent samples was measured using the Shannon diversity index . The comHalospirulina was observed to be the most abundant in the air vent samples of two HNM  data, a total of 58 and 53 bacterial genera were identified from the air vent and ground surfaces of both HNM and HVM and presented in le of H8 B. The geHNM H12, C. Howeveacterium D.Saccharomyces in H17 and Phaeoisaria in H6 . Similarly, no significant difference was observed among bacterial communities in ground samples collected from HNM and HVM (p = 0.51). No significant differences were observed in the ground dust samples from HNM and HVM (p = 0.60). However, a significant difference in fungal communities in air vent samples of HNM and HVM was observed (p < 0.05). Along with house types, the difference in the composition of bacterial and fungal communities in ground and air vent samples was also calculated. The difference in bacterial communities between ground and air vent samples in HNM . Similarly, the difference in fungal communities between ground and air vent samples in HNM and HVM was presented in p < 0.05 level .To visualize the differences in taxonomic compositions between fungal and bacterial communities, we generated NMDS ordination plots , which ps in HNM E and HVMs in HNM F showed Alternaria alternata was recorded as the most abundant fungal species in ground and air vent dust samples from both HNM and HVM, with some exceptions. For example, it was not detected in the ground sample collected from H9 or the air vent samples collected from H1 and d H5. Aspergillus niger and Aspergillus versicolor were also found in some houses’ ground and air vent samples; however, Aspergillus sydowii was not recorded in any of those houses.The presence and absence of a selected group of fungi known to be associated with respiratory complications ,40,41 ha2 concentration, temperature, and relative humidity. The relationships were examined using Spearman correlations for ground  is a fungus with type I and III allergens associated with skin and nail infections in humans and infrequently causes cerebral and systemic infections in immunocompromised individuals [Alternaria, Pseudopithomyces, and Cladosporium were the dominant fungus genera in an indoor environment, which is consistent with our finding [Alternaria, Cladosporium, and Leptosphaerulina (Pithomyces) are common outdoor fungi; thus, they may have originated and been transmitted from outdoor to indoor environments. Interestingly, in our metataxonomic analyses, the abundance of Penicillium was found to be low, which is contrary to several previous reports showing Penicillium as one of the most abundant airborne fungal genera in the indoor air of moldy homes [Penicillium brevicompactum in the ground and air vent samples from at least one moldy home, while air vent samples from two moldy homes yielded positive results for Penicillium citrinum. The low detection rates of Penicillium could be attributed to the assay design (NGS versus qPCR), differences in environmental conditions, or other unknown factors.The most abundant fungal genus in all these houses was found to be diseases ,63. It mthomyces , is commthomyces . Similarmunities . On the erbation ,68,69, wividuals ,71,72,73 finding . Alternady homes ,74,75. HThe NMDS gradient analysis produces ordinations based on the dissimilarity matrix, or distance between different data sets. In this investigation, the NMDS analysis  revealed2 level, humidity, and temperature, using Sperman’s Rho statistics. Depending on the house status (HNM and HVM) and sampling site (ground and air vent), fungal communities exhibited some positive and negative relationships; however, none of these correlations were statistically significant (p < 0.05). This could be due to a number of factors, such as sample size, variability in environmental conditions, or other unknown factors that may have influenced the results. Environmental factors are known to be associated with asthma and other respiratory illnesses [The diversity of microbial communities in an indoor environment highly depends on the building’s occupants and the surrounding outdoor environment . There illnesses . To thisllnesses ,80.Alternaria alternata, followed by Chaetomium globosum. The spore produced by A. alternata is a well-known biological contaminant and has been reported to be an inducer of several respiratory illnesses [Chaetomium globosum is a mesophilic mold ubiquitously distributed in nature and was reported to be associated with onychomycosis [Chaetomium abundance in both house types (moldy and non-moldy) in the present study may indicate the existence of dampness on gypsum boards or plywood wallboards. Furthermore, Aspergillus versicolor was also detected in many homes (both HVM and HNM), which is associated with several health problems, including neurological issues and pulmonary infections [Exposures to pathogenic fungi, including molds, are reported to be associated with several health problems, such as allergies, shortness of breath, chest tightness, and other respiratory complications ,41. Usinllnesses . Chaetomomycosis . This fiomycosis ,84. Therfections ,86. Intefections ,57.n = 28 homes) [The limitations of the present study may include a lack of statistical power as we only sampled from a total of twenty homes. Nonetheless, there are reports of similar studies with a limited sample size (such as 8 homes) . A large8 homes) .In this study, we used molecular methods to report the bacterial and fungal communities, the presence of fungal species known to be associated with respiratory diseases in HNM and HVM, and their correlations with indoor environmental factors. We found that these molecular methods provide rich information and are very useful in fungal and bacterial diagnostics in indoor environments. Future research aimed at assessing indoor air quality in homes will need to use updated technologies to ascertain the correlation between the microbiota of the built environment and adverse health effects.


Neutron capture therapy, or NCT, is a unique irradiation-based method that is used to treat invasive advanced tumors as it minimizes the impact on healthy cells and tissues. Modern neutron sources for NCT include accelerators that can be installed in treatment facilities. One of the main conditions for the successful application of NCT is sufficient accumulation of the drug in tumor cells which allows it to capture neutrons, be activated, and release energy that destroys cancer cells. Among the elements capable of capturing neutrons, boron, gadolinium, and lithium are considered to be the most suitable for NCT. In our study, we showed the results of neutron capture therapy with gadolinium in a group of dogs and cats with spontaneous tumors to clarify the technical aspects of the method while adjusting the treatment conditions as close as possible to those of clinical trials.®, 0.6 mL/kg b.w.), was used. We observed mild and reversible toxicity related to the treatment. However, no significant tumor regression in response to the treatment was observed. In most cases, there was continued tumor growth. Overall clinical improvement after treatment was only temporary. The use of Gd-DTPA for NCT had no significant effects on the life expectancy and quality of life of animals with spontaneous tumors. Further experiments using more advanced gadolinium compounds are needed to improve the effect of GdNCT so that it can become an alternative to boron neutron capture therapy. Such studies are also necessary for further NCT implementation in clinical practice as well as in veterinary medicine.We conducted a clinical veterinary study on neutron capture therapy (NCT) at a neutron-producing accelerator with seven incurable pets with spontaneous tumors and gadolinium as a neutron capture agent . Gadolinium-containing dimeglumine gadopentetate, or Gd-DTPA (Magnevist Certain malignancies are extremely difficult to treat and can still remain lethal following treatment due to both their invasive nature and the inability of all modern treatment modalities to influence single tumor cells that migrate into healthy tissues and lead to the development of new tumor masses at a distance from the initial tumor that seemed to be cured. This may happen in the case of malignant melanoma with distant metastases, malignant glioma with fast local recurrence, or another type of invasive cancer that leaves no chance of survival despite the best efforts of modern medicine. Therefore, the search for more effective cancer therapies, including more advanced irradiation methods capable of targeting single tumor cells, is still ongoing.10B)-based compounds have already been tested clinically, with one of them, boronophenyl alanine (BPA), finally approved for clinical application in Japan [157Gd) possesses the best ability to capture neutrons and releases gamma rays that ionize matter in tumor cells [6Li) used as a neutron capture compound for NCT have been recently initiated, and additional details related to its application are yet to be discovered [The principle of low-energy neutron capture by a drug followed by a local release of energy that destroys tumor cells has been employed in neutron capture therapy (NCT), which is a unique and sometimes the only applicable treatment for certain types of malignancies. The main advantage of NCT is its selectivity such that only tumor cells are affected and healthy tissues are preserved due to the precise intracellular accumulation of a compound that captures neutrons, decays, and produces energy that disrupts tumor cell DNA integrity ,2,3. Sevin Japan ,5. The gor cells . Experim10B, a concentration greater than 20 µg/g in tumor tissue is needed to achieve a therapeutic effect [10B has a smaller neutron capture cross-section and the energy released due to its interaction with a neutron is limited by cell size, which plays a key role in treatment selectivity. Clinical trials on NCT at nuclear reactors using 10B  have demonstrated effectiveness in treating different malignancies [10B compounds to accumulate in liver metastases of colorectal cancer [In the case of c effect . Accordignancies ,12,13,14gnancies ; Kato etgnancies  and Kankgnancies  and Miya10B-containing compounds in research and clinical practice. Therefore, Gd-based drugs, which are already registered in many countries and used clinically for purposes other than NCT, could be alternatives to 10B compounds.However, the efficacy limitations of previously and currently used boron compounds, the complexity of the boron enrichment process required for their production, and their high cost restrict the use of ®, became one of the first contrast agents to be used in clinical trials and was originally approved by the FDA for clinical use in 1988 [157Gd/g can be achieved in brain tumors with an MRI contrast agent such as Gd-DTPA when injected intravenously at 0.5 mmol/kg body weight [Thus, beginning in 1985, dimeglumine gadopentetate (Gd-DTPA), or Magnevist in 1988 ,16,17. I in 1988 ,19,20. Iy weight . The effy weight ,27,28,29® by Sumitomo Heavy Industries, Ltd. , based on a cyclotron accelerator operated at an energy level of 30 MeV with a current of 2 mA and a beryllium target, which is already installed in clinical treatment facilities in Japan [Regarding the irradiation facility for NCT, a specialized nuclear reactor channel or a charged particle accelerator is needed for neutron generation ,31,32,33in Japan ,32. A tain Japan .® containing boron phenylalanine (BPA) was approved for head and neck cancer treatment in adults, with national insurance coverage [In Japan in 2020, accelerator-based BNCT using the drug Borofalancoverage ,5,44. Thcoverage ,44,45,46In many countries, preclinical radiobiological studies are required prior to treating human malignancies with accelerator-based NCT. In this case, studies using cells and animal tumor models play a key role ,47. HoweIn this case, enrollment of large animals of the mammal family, particularly domestic animals, such as cats and dogs, is the most suitable for estimating the radiobiological effects of NCT. Such animals have a large body mass, which allows for the selection of dosimetry parameters similar to those in humans. In addition, malignant tumors in large animals are easy to examine morphologically. The localization and nature of spontaneous tumors in domestic animals are similar to human tumors in many ways. Accordingly, the applied surgical treatment, chemotherapy, and radiation therapy methods show similar results. Since malignant tumors are a common cause of death in pets  and stan® (dimeglumine gadopentetate), and a neutron beam obtained at the accelerator developed at the Institute of Nuclear Physics in Novosibirsk, Russian Federation [NCT is designed to be applied mainly in head and neck cancer, intracerebral tumors, melanoma, and metastatic lesions, which are very common in pets. We are the first to conduct a preclinical GdNCT effectiveness study in large mammals with malignant tumors using a registered gadolinium-containing MRI contrast agent, Magnevistderation  with irrderation .Thus, in this study, we aim to investigate the effect of GdNCT to increase the knowledge of NCT and optimize its therapeutic effectiveness. This study on gadolinium neutron capture therapy was performed based on ethical standards and approved by the Ethics Committee of the Institute of Cytology and Genetics, Novosibirsk, Russian Federation. In addition, the informed consent of the animal owners for this type of therapy was obtained in each case. For this study, we selected animals with spontaneous tumors in satisfactory somatic condition with the possibility of anesthesia and clinical observation for more than three months or those with tumors that were difficult or impossible to remove surgically, taking into account complications. We also included animals with tumors that had relapsed after surgical treatment or previous chemotherapy. A total of seven animals (five cats and two dogs) received GdNCT—the data are presented in Before treatment, a computed tomography (CT) scan was performed to visualize tumor location, size, and possible metastases. The imaging data were used for treatment planning. Before the GdNCT session, the neutron spectrum and the absorbed dose depth distribution in the simulated tumor, skin, and surrounding tissues were calculated. To calculate the minimum effective dose with a median of 30 Gy-Eq in the tumor, a tumor/surrounding tissues ratio of 3:1 was used, with an assumed average gadolinium concentration of 5.5 mg/g in the tumor at the time of irradiation and 1.8 mg/g in the surrounding tissues. The tumor-to-blood ratio was assumed to be 1:1. The upper dose limit for the skin was 18 Gy-Eq and 12 Gy-Eq for the mucosa [The irradiation settings included a proton energy level of 2.05 MeV and an integral current between 2.2 and 4.4 mAh, depending on the presence of previous radiation therapy, tumor volume, and localization. The average irradiation session lasted for 2 h.®), manufactured by Bayer AG , was administered fractionally at the maximum dose for humans recommended by the manufacturer (0.6 mL/kg). This was accomplished via an intravenous injection immediately before the radiation (0.3 mL/kg) and another injection (0.3 mL/kg) in the middle  of the radiation session. Animals received intravenous anesthesia using a standard dose of Dexdomitor, dexmedetomidine plus Zoletil 100, or tiletamine plus zolazepam. Dimeglumine gadopentetate . The average blood concentration of gadolinium was 182 ± 217 ppm. Since this study was also performed to infer how much gadolinium accumulates within tumors, the amount of gadolinium in the tumors was predicted using an assumed tumor-to-blood ratio of 1:1 and a tumor-to-surrounding tissues ratio of 3:1.Animal positioning during therapy, collection of blood samples for analysis, and post-treatment monitoring of the animals for three months were performed according to the methodology of the previous study .Animals (cats and dogs) are described in the sequence of their examination and treatment with GdNCT.Age: 11 years.Symptoms: worsening of nasal breathing and nasal discharge.Tumor localization: right nasopharyngeal area. The nasopharyngeal tumor was visualized using CT.Treatment other than GdNCT : none.Final pathological diagnosis: highly differentiated adenocarcinoma of the nasal mucosa.The course of the disease after GdNCT: during follow-up, the overall condition was satisfactory, and nasal breathing improved. At the examination, the visible part of the tumor decreased in size. A CT scan performed 3 months after GdNCT revealed no significant changes compared to the pretreatment visualization .Age: 14 years.Symptoms: impaired nasal breathing and nasal discharge.Localization and size of the tumor: nasal area; according to CT, the tumor size was 32 × 39 × 47 mm.Treatment other than GdNCT: surgical removal of the nasal tumor, twice.Final pathological diagnosis: chondrosarcoma.The course of the disease after GdNCT: after one month of follow-up observation, the overall condition was satisfactory. However, starting from the second month, a gradual deterioration of the condition, nasal bleeding, difficulty in breathing, and decreased appetite were noted .Age: 6 years.Symptoms: impaired nasal breathing and nasal discharge.Tumor localization and size: left nasal area with bone destruction and regional lymphadenopathy. According to the CT scan, the tumor size was 33 × 26 × 26 mm.Treatment other than GdNCT: standard X-ray radiotherapy with doses of 35.4 Gy for the tumor (6 sessions) and 30 Gy for the lymph nodes (6 sessions). The outcome of that treatment was tumor recurrence.Pathological diagnosis: squamous cell carcinoma.The course of the disease after GdNCT: after irradiation, the animal was lethargic and had facial edema and a necrotic area (circled in the photo) with exudate. Two months after GdNCT, there were signs of tumor recurrence in the back of the nose, upper jaw, and corner of the eye .Age: 11 years.Symptoms: gingival swelling and ulceration, bleeding, decreased appetite, regional lymphadenopathy, and lacrimation from the right eye.Tumor localization and size: oral cavity, upper jaw on the right side, with metastasis to the submandibular lymph node. Based on a head CT, the tumor size was 25 × 40 × 15 mm.Treatment other than GdNCT: none.Pathological diagnosis: highly differentiated squamous cell carcinoma.The course of the disease after GdNCT: the animal’s condition and appetite after GdNCT were normal, and there was a visual reduction in tumor size during the early observation period. There was no discharge. After irradiation, the eye continued to tear. Twenty-three days after GdNCT, the condition worsened. The tumor began to disintegrate, the gum was covered with bloody ulcers, and holes appeared in the cavity; the cat also experienced spittle vomiting and black liquid stools, a decrease in appetite, and would rub the tumor site. After 42 days of treatment, the tumor had not become visually smaller, the gum was bleeding, the animal’s appetite was not quite stable, and constant use of analgesics was necessary. Three months after GdNCT, active tumor progression was found at the examination .Age: 13 years.Tumor localization and size: basal neoplasm of the middle lobe of the right lung with involvement of the cranial and caudal lobes and single metastases to the lungs and regional lymph nodes. According to the CT scan, the tumor size was 33 × 35 × 42 mm.Treatment other than GdNCT: none.Pathological examination was not done.The course of the disease after GdNCT: general improvement, breathing recovery, and right lung excursions one month after treatment. Activity and appetite improved. According to the CT scan, 2 months after GDNCT, the mass decreased in size to 32 × 38 × 36 mm .Age: 10 years.Symptoms, localization, and size of the tumor: soft tissue mass in the upper jaw on the left side with invasion into the nasal cavity and left orbital space. Destruction of the upper jaw bone. According to the CT scan, the tumor size was 16 × 31 mm.Treatment other than GdNCT: none.Pathological diagnosis: squamous cell carcinoma.The course of the disease after GdNCT: the cat’s appetite improved, their eye was open a little, and their activity was unchanged. Sneezing disappeared after irradiation but then resumed at a lower frequency than before irradiation; grunting sounds (which were present before irradiation) also disappeared. The overall condition improved. One month after GdNCT, the condition worsened, with the tumor size increasing to 38 × 59 mm, according to CT. The condition further deteriorated, and subsequently, the animal died .Age: 6 years.Localization and size of the tumor: right femur and iliac bone; according to a CT scan, the tumor was 34 × 32 × 37 mm in size.Treatment except for GdNCT: four courses of chemotherapy.Pathological diagnosis: osteogenic sarcoma.The course of the disease after GdNCT: on the tenth day after NCT, the animal was eating, drinking, and feeling well. A CT scan three months after irradiation revealed an increase in the tumor size to 58 × 38 × 46 mm .Previously, our research group conducted studies on BNCT in laboratory animal tumor models and spontaneous tumors in pets (cats and dogs) using BSH ,50,51.157Gd in NCT has been limited to cell experiments and small animal studies so far. Despite the advantages of gadolinium, such as a high gradient of accumulation ratio in tumors relative to healthy tissues, the large cross-section for thermal neutron absorption, and the possibility of using highly efficient Auger electrons from gadolinium to destroy tumor cells, there are several drawbacks associated with its use. During neutron capture by gadolinium, the most important issue is the effect of the generated gamma radiation on the healthy tissues surrounding the tumor. This radiation possesses high energy  and travels a considerable distance through biological tissues, which might cause irradiation of healthy cells and an insufficient irradiation dose to the tumor. Electrons and X-rays account for a small fraction of the energy released in a Gd reaction, which is 63 keV. However, these electrons and X-rays can play a significant role in affecting the tumor, with their specific effects depending on the tumor’s size and the gadolinium location in relation to the tumor cell nuclei. Initially, many studies considered only the dose distribution from gamma radiation from Gd reactions. However, Martin et al. (1989) and Laster et al. (1996) found that in their experiments with cells, the effect of gadolinium-related neutron capture reactions was more significant than identified in previous studies where it was associated only with gamma radiation [BSH and BPA are standard compounds typically used in BNCT. They are expensive and difficult to obtain in some countries. For example, a single clinical NCT session might require a volume of the drug that costs about 30,000 USD in equivalent. Since gadolinium compounds are currently used in clinical practice for contrast enhancement during MRI, such agents represent perfect theranostic drugs capable of combining NCT and MRI. Although this is an advantage of GdNCT, the use of adiation ,53. In tadiation .®, there is a retention of gadolinium in the glial tumor, which is already clearly visible during the visual analysis of MR images, and at the same time, there are no changes in the intensity of the surrounding white matter. The ratios between the transport coefficients of Magnevist® in the directions from blood to tissue and from tissue to blood were calculated. For glial tumor tissue, this ratio was 9.5–10, i.e., the reverse transfer was slower by one order of magnitude. Obviously, this situation will ensure sufficient retention time of Magnevist® molecules in the tumor tissue with negligible accumulation in normal tissues, which is the key to successful NCT with selective tumor cell targeting [Karpovich et al. proposed a method of therapy planning for GdNCT that relies on quantitative assessment of the content of Gd (III) complexes in the tumor and surrounding tissues based on a multicompartment phenomenological pharmacokinetic model adapted to real patient study data, with the possibility of visual assessment by MRI . In patiargeting . These rargeting .Despite promising dosimetry data, the efficacy of Gd-DTPA and Gd-DOTA as therapeutic agents for GdNCT was initially predicted to be low due to a potentially insufficient accumulation of gadolinium in tumor cell nuclei. However, De Stasio et al. showed that Gd-DTPA penetrates cancer cell nuclei in vitro, which led to a reconsideration of the GdNCT approach using that compound . In vivoTo increase the effectiveness of Gd-DTPA in GdNCT, continuous intra-arterial infusions have been proposed . In a st® has been presented [A proof of principle for cell death by Gd neutron capture in U87 glioblastoma multiforme (GBM) cells preloaded with Magnevistresented . Later, resented .® have also been studied. Brugger et al. (1989) published one of the first articles reporting on Gd chelates used as NCT agents [®) as an agent for neutron capture therapy [Gadolinium NCT agents that are potentially more effective than MagnevistT agents . The posT agents . Matsumu therapy . Gd-BOPTDifferent researchers suggested using liposomal compositions or chitosan to increase the concentration and long-term deposition of gadolinium in tumors ,27,29. L®) in human malignant fibrosis histiocytoma (MFH) cells [Fujimoto T. et al. (2009) used gadopentecic acid (Gd-DTPA) with chitosan to develop gadolinium-loaded chitosan nanoparticles (Gd-nanoCPs) and studied their accumulation and MR imaging visualization enhancement effect compared to Gd-DTPA (MagnevistH) cells . The autH) cells . In a laH) cells . In vitrH) cells . Other aH) cells .® and BPA were used. In the GdNCT group, 70% of cases achieved complete regression of the tumor within 30–45 days; in 14% of the animals, recurrence occurred after one month, and life expectancy was between four and six months. In the BNCT group, complete tumor regression was achieved in 95% of cases; in 20% of cases, recurrence occurred after three to four months, and the life expectancy was approximately eight months. In the gamma therapy group, 90% of the dogs showed tumor recurrence one month after treatment, metastases after 2–3 months, and a life expectancy of approximately three months. Regarding studies on neutron capture therapy in large animals using gadolinium compounds, a group led by Professor Mitin treated dogs aged between 9 and 15 years with histologically confirmed diagnoses of oral mucosal melanoma . Dipenta® drug instructions, we used the maximum recommended dosage of 0.6 mL per kg of body weight or 0.3 mmol per kg of body weight, which in a proportional ratio should provide up to 180 µg of 157Gd/g in tumor tissue. We injected the drug twice at half the dose calculated per body weight to ensure a homogeneous concentration of the drug in the tumor for the entire duration of the GdNCT session.In our study, according to the MagnevistDespite meeting all the conditions for maximum accumulation of the drug in the tumor tissue, which suggests a maximum NCT effect, we observed negative changes that manifested as increases in tumor volume and clinical deterioration during the follow-up period one to three months after GdNCT in most of the animals. We observed a short-term absence of disease progression only in some cases; there were also no adverse reactions of the skin or fur at the irradiated sites. Thus, we could not show a significant positive effect of NCT for spontaneous malignant tumors in domestic animals. Computed tomography performed three months after GdNCT showed no changes or increases in tumor volumes, indicating an insufficient tumor response in all cases. Survival with poor quality of life was observed up to three months after irradiation, and the survival rate and quality of life did not improve over time. Typically, within two weeks (14 days) after GdNCT, the initial positive treatment response was replaced by a deterioration of the overall condition and quality of life. No change in tumor size after therapy and continued tumor growth could be related to the spread, depth, and infiltrative nature of spontaneous tumors as well as an insufficient concentration and rapid washout of Gd-DTPA from tumor tissues despite two injections during irradiation sessions, the extracellular localization of Gd-DTPA, and the presence of Gd-DTPA mainly in the tumor periphery. As a result, large tumors received insufficient radiation doses in the central and deeper parts. Thus, the small number of animals, different morphological characteristics of the tumors, and the lack of computed tomography data in some cases may represent the current study’s limitations.A solution to improve the therapeutic outcome at this stage would be using gadolinium drugs that can accumulate inside tumor cells, performing several GdNCT sessions, and combining GdNCT with other treatment modalities. Therefore, it is obvious that new gadolinium-containing agents capable of selective accumulation in tumors are required for GdNCT. Based on recent publications, one of the most promising theranostic agents is AGuIX, a radiosensitizer and MRI contrast agent with a high gadolinium concentration in its structure . It has Neutron capture therapy often remains the last option for the treatment of spontaneous tumors in animals, while other methods prove to be ineffective. Studies on large animals allow for treatment conditions to be adjusted as close as possible to those of clinical trials and help clarify the technical aspects of NCT. Using a unique accelerator source for neutron generation, as done in these studies, represents a pioneering application of the method worldwide. The combination of gadolinium administration and accelerator-based neutron irradiation provides additional data on the current conditions of GdNCT. These data can then be compared with those obtained by scientific groups conducting BNCT and other methods of radiation therapy for malignant tumors. At this stage, it is evident that successful GdNCT requires new gadolinium-containing drugs capable of sufficient tumor accumulation. Furthermore, the results of preclinical studies in large animals will allow us to prepare for the initiation of appropriate clinical trials.


Parenting was drastically challenged during the COVID-19 pandemic as families complied with the consequent containment for sanitary and social reasons. The purpose of this study is to explore the relationships among self-report measures  and the employment condition  that might be associated with the everyday life change, and their contribution towards family’s daily routines during the first COVID-19 lockdown. A sample of 878 parents (90.2% mothers), aged from 21 to 61 years (39.58 ± 6.08), and mainly of Portuguese nationality (97.2%), was gathered through an online survey. Participants completed sociodemographic data and answered questions related to four positive parenting dimensions  rating on a five-point Likert scale how much their behavior changed during the lockdown. Descriptive statistics and multilinear regression models were tested as well as a mediation model. The results showed that Emotional experience, Co-parenting, and Support network played an important role in family Daily routines, reinforcing that the first COVID-19 lockdown was lived differently by parents according to their employment conditions. Furthermore, the pathways model of factors associated with parental adaptability is suggestive that the direct effect of Emotional experience on Daily routines is partially mediated by Co-parenting and Support network. In line with previous studies on the complexity and dynamic ecology of parenting, the reported indicators showcase the interrelated facets of parenting and its corresponding support needs. Further, clear indicators for the design of family support interventions and family resilience enhancement to stressor events are discussed. Parenting is a challenging task, especially in times of crisis when families faced the COVID-19 public health threat worldwide. However, above and beyond the turmoil, the lived changes that the pandemic evoked also capitalized families’ resources and, ultimately, coping with the strain and stress promoted their resilience. In the perspective of Froma Walsh .According to Rodrigo , parentiWorldwide, published studies about parenting during the COVID-19 pandemic illustrate that parents and children’s personal and dyadic processes as well as all-enveloping external factors contribute to adaptative changes on parenting and family dynamics. In fact, the pandemic experience is often related as a moment of crisis  but also as a moment of opportunities  . Yet, stAlong with parental self-regulation variables, the impact of the lockdown on Portuguese children was also associated with parental competency perceptions and parent–child interactions . Yet, inApplying the systemic approach to address changes in parenting during COVID-19 pandemic, the purpose of this study was to deepen our understanding regarding the associations among the parent’s emotional experience, parenting daily routines, co-parenting and support network. Additionally, in line with the already published study, a secondary goal was identifying the different employment conditions associated with everyday life changes in parenting dimensions  and their contribution towards family’ daily routines during the first COVID-19 lockdown. Specifically, the hypotheses tested in this study consider the following assumptions:Hypothesis 1.Positive changes in the parent’s emotional experience  would be associated with more positive changes in daily routine .Hypothesis 2.Positive changes in co-parenting and support networking would enhance the parent’s emotional experience, yielding a mediational effect on parenting adaptability in the daily routine.The present study is part of a Portuguese study (Parenting in COVID-19 times) of which some results have already been published . The desThe participants were selected from a larger sample of parents (1384 parents), and they were selected because they were the group of parents who manifested more changes in family dynamics during the period of the first COVID-19 lockdown according to a previous study [So, for the main purpose of this study  we selected these parents because they revealed more changes in their family dynamics , and we Also, to be included in the study, participants were either a parent or a substitute caregiver of one or more children 18 or under, living at home with them.An ad hoc questionnaire was used to collect the data . The qu(a)The Daily routine scale (15 items) assesses the degree of change on basic care in daily chores . This scale has four subscales: positive discipline ; parent–child activities ; nurturing ; and enriched environment . For this sample, the Cronbach’s alpha was 0.815 and McDonald’s omega was 0.815.(b)The Co-parenting scale (11 items) assesses marital involvement and mutual support in multiple childcare duties and has two subscales: parental alliance  and parental agreement . For this sample, the Cronbach’s alpha was 0.884 and McDonald’s omega was 0.886.(c)The Emotional experience scale (14 items) assesses emotional experience during the lockdown period and has four subscales: emotional sensitivity ; emotional tension ; emotional regulation ; and stress management . For this sample, the Cronbach’s alpha was 0.816 and McDonald’s omega was 0.788.(d)The Support network scale (nine items) assesses the changes in the support of formal and informal networks provided to the family during lockdown and has two subscales: informal network  and formal network . For this sample, the Cronbach’s alpha was 0.843 and McDonald’s omega was 0.829.The four scales  are briefly described and the Cronbach’s alpha and the McDonald’s omega, which were calculated for this sample, are also presented :(a)The The responses of the four scales were rated on a 5-point scale ranging from 1 to 5, reflecting the degree of change . A score of zero was included to allow the participants to check whenever the item(s) did not apply and these values were coded as missing.The present study is part of a Portuguese study (Parenting in COVID-19 times) that was approved by the Ethics Commission for Research in Social and Human Sciences of the University of Minho (reference CEICSH045/2020 of 13 May 2020).To perform the study, an ad hoc questionnaire was constructed and made available through an online platform. At the beginning of the questionnaire, the participants acknowledged the purpose and procedures of the study, the assurance of confidentiality and voluntary participation, and their consent to participate in the study was mandatory to continue the filling of the questionnaire.Parents were recruited trough social media platforms like Facebook and WhatsApp.The study was conducted by asking the participants about to the first lockdown in Portugal . The data were collected from 15 May to 26 June, at the end of the academic year while several restrictive measures, including lockdowns, remained in force.2).Statistical analyses were conducted using SPSS, version 28.0  and JASP 0.16.0.0 for Windows. The internal consistency reliability study was done through analysis of the Cronbach’s alpha and McDonald omega ,20. DescThe mediation model was calculated using the statistics program JASP version 0.16.0.0. Emotional experience was included as a predictor, social support and co-parenting as the mediators, and daily routine as the outcome. Parent’s employment was not included because it was a categorical variable. Direct, indirect, total indirect and total effects were calculated with bootstrap confidence interval based on 1000 resamples.p < 0.05.The level of significance was set to The participants in the study were 878 parents (90.2% mothers), with ages from 21 to 61 years old  and were mainly of Portuguese nationality (97.2%).Most of the participants was married or had a stable relationship (78.2%) and had a university degree (79.7%). The majority had one child (41.2%) or two children (46.8%) living at home (the number of kids ranged from one to five). The age of the children was between one year old to 18 years old  and most of the children revealed no special educational needs (88%).The participants experienced on average 66.4 days of lockdown, ranging from zero to 122 days in lockdown (SD = 15.1).All parents of this sample were employed, but a group of them were at home teleworking (67.7%) and the other group was at home, on a licensed work leave and receiving governmental assistance (32.3%), to care for their children.p < 0.01), parental agreement , parental alliance , emotional experience , emotional sensitivity , emotional regulation , support network  and informal network .The results showed moderate positive correlations for daily routine and co-parenting , parental agreement , parental alliance , emotional experience , emotional sensitivity , emotional regulation  and informal network .Moderate positive correlations were found for positive discipline and co-parenting , parental agreement , emotional sensitivity  and emotional regulation .Moderate positive correlations were found for parent–child activities and co-parenting , emotional sensitivity , and emotional regulation .Moderate positive correlations were found for nurturing and emotional experience , parental agreement , emotional experience , emotional sensitivity , and emotional regulation .Moderate positive correlations were found for enriched environment and co-parenting .p < 0.001). The results indicated that the model could explain 31.4% of the positive discipline changes . Parental agreement , emotional sensitivity , emotional regulation  and informal network  influenced the changes in positive discipline. More changes in parental agreement, emotional sensitivity, emotional regulation and informal network produced more changes in positive discipline.For positive discipline , the modp < 0.001). The results indicated that the model could explain 22% of the parent–child activity changes . Employment condition situation , parental agreement , emotional tension (β = −0.169 p < 0.001) and emotional regulation  influenced the changes in parent–child activities. At-home parents with government licensed work leave, more changes in parental agreement and emotional regulation and less changes in emotional tension produced more changes in parent–child activities.For parent–child activities , the modp < 0.001). The results indicated that the model could explain 17.6% of the nurturing changes . Parental agreement , emotional sensitivity , emotional regulation  and stress management  influenced the changes in nurturing. More changes in parental agreement, emotional sensitivity and stress management produced more changes in nurturing.For nurturing , the modp < 0.001). The results indicated that the model could explain 22.2% of the enriched environment . Employment condition , parental agreement , emotional sensitivity , emotional regulation  and informal network  influenced the changes in enriched environment. At-home parents with government licensed work leave, and more changes in parental agreement, emotional sensitivity, emotional regulation, and informal network produced more changes in enriched environment.For enriched environment , the modThe hypothesized mediation model that was constructed, and consequently tested, revealed a direct effect of the Emotional experience in Daily routines, and Support networks and Co-parenting as partial mediators between Emotional experience and Daily routines.2 = 0.07) for support network and 11.4% (R2 = 0.114) for co-parenting. The total effect is 0.43 (p < 0.001), the direct effect is 0.24 (p < 0.001) and the total indirect effect is 0.19 (p < 0.001). In addition, both indirect effects are statistically significant: emotional experience → support network → daily routine and emotional experience → co-parenting → daily routine. Support network and co-parenting are partial mediators between emotional experience and daily routine.The results of the mediation of co-parenting and support networking between emotional experience and daily routine are described in The main purpose of this study was to analyze the relations among the different factors associated with adaptative parenting . The second aim resulted from an extension of previous findings with this group of parents seeking to identify whether the impact of the parents’ employment condition was also perceived in daily routine dimensions, namely predicting the changes in parenting adaptability during the first COVID-19 lockdown. As can be seen, the results allowed us to state that the aims of the study were achieved and its main contributions are discussed next.Regarding the first purpose, the results revealed a rather homogenous and stable pattern of association among daily routine changes with co-parenting changes and emotional experience. These changes are extensive to positive discipline, parent–child activities, nurturing and enriched environment. In common, the four sub-dimensions of daily routine indicate associations with observed changes in the parental agreement dimension and emotional regulation. Indeed, no matter the singularity of supplementary associations with other sub-dimensions of co-parenting  and emotional experience , the predominance of this pattern presents an effortful conscience of balance and control needs, which is pivotal to parental functioning in competent ways. These findings align with other studies where couple relationships changed during the pandemic and were considered a key factor to discuss parenting and family dynamics, excepting single-parent families ,11,12,22Additionally, daily routine was associated with the total support network and, more specifically, to an informal network, which was associated with positive discipline and an enriched environment. Conversely, formal network was not associated with the daily routine scale nor with its subscales, possibly reflecting the social distancing to services and professionals. Nonetheless, this result might be detailed across different types and sources of formal support other than those directed to schools and neighborhoods . HoweverContrary to the previous findings regarding the impact of the employment condition on parenting dimensions, these results were not confirmed in the present study . This miSecondly, regression models enabled to test the differential effect of parental agreement and emotional regulation on the daily routine subscales. The findings indicated the importance of parental agreement and emotional regulation as predictors of positive discipline, parent–child activities, nurturing and an enriched environment. The observed causality of parents’ psychological characteristics and relational resources to parenting in daily life is a trace of the adaptative changes in different aspects of daily routines, and its critical role in a balanced and affectionate family’s functioning as well as for the parents and children’s well-being. Yet, regardless, it is possible to disentangle the weight and direction of specific factors—for instance, that emotional sensitivity predicts changes in positive discipline, nurturing and enriched environment, that emotional tension and emotional regulation impact parent–child activities, and that stress management predicts nurturing—these different predictors gain significance and expression in the whole context of good parenting. And the same can be said for other proximal or distal influencing factors. As shown, informal network was a predictor of positive discipline and enriched environment, while endorsement of new parent’s employment policy in COVID-19 times had an impact on parent–child activities and enriched environment. In the parenting process, these enhancing factors are clearly underlined by Rodrigo . HighligThese results might outline an informative agenda when designing and programming family interventions, considering the nature and essence of parenting. Not solely the array of domains but the complexity and the dynamics of adaptative processes provide insightful recommendations for psychoeducational interventions and promotion of positive parenting and family resilience. Also, departing from lived experiences and concrete family situations in different countries and social realities might be suggestive of more specific and responsive interventions according to parents’ needs. Interventions that help the development of positive parenting and family resilience are crucial  as well Regarding post-pandemic interventions, these remain important as a societal and intergovernmental approach to face the consequences of the pandemic, strengthening the intersectorial delivery and developing their progressive universal delivery without compromising the implementation of specific measures for at-risk populations . LookingDespite the contributions of this study, some limitations must be acknowledged. The instrument used to collect data was a self-report questionnaire, depending on personal perceptions. Data were collected online and the conditions about the questionnaire filling could not be controlled. Furthermore, participants did not represent the Portuguese population and generalization of the results is limited. Most participants were mothers, and their expectancies and attitudes facing motherhood, which were not measured, could influence the personal perceptions of changes and, consequently, the mediation model . Also, aNonetheless, and to conclude, this study made contributions to the knowledge in the recent and novel condition related to COVID-19 lockdowns and families. We think the results might enlighten present and future studies, and interventions, dedicated to remedy, and to prevent, family crisis arising from stressful events such as the COVID-19 pandemic.


Background: Multiple sclerosis (MS) patients present dyslipidemia and functional disability. Epigallocatechin gallate (EGCG) and coconut oil have been shown to be effective against dyslipidemia. Objective: To analyze the relationship between lipid profiles, fat consumption, and functional disability in patients with MS after administering EGCG and coconut oil. Methods: A four-month pilot study was conducted on 45 MS patients, divided into an intervention group (IG) and a control group (CG). The IG received 800 mg of EGCG and 60 mL of coconut oil. Lipid profiles were measured before and after the intervention, along with other data such as dietary habits, inflammatory markers, and functional capacity. Results: Dyslipidemia did not correlate with the patients’ fat consumption. After the intervention, triglycerides (TG) levels were lower in IG compared to CG. This decrease was positively correlated with an improvement in functional disability ) and negatively with high-density cholesterol (HDL) and apolipoprotein A1. Significant and positive correlations were observed between EDSS and C-reactive protein (CRP) in the IG. These changes in the IG could be related to body fat decrease, whose percentage shows a positive correlation with CRP and TG levels, and a negative correlation with HDL levels. Conclusions: Patients with MS present a certain type of dyslipemia not associated with their nutritional habits. The administration of EGCG and coconut oil seems to decrease blood TG levels, which could explain the functional improvements. Multiple sclerosis (MS) is a chronic, neurodegenerative, and inflammatory disease characterized by a loss of myelin in the neuronal axons . ClinicaThere is currently no cure for MS and different nutrients or antioxidant molecules stand out as possible alternatives that may reverse or reduce the progression of the disease. Among these, the polyphenolic antioxidant epigallocatechin gallate (EGCG) has achieved multiple benefits in neurodegenerative diseases models such as Alzheimer’s and Parkinson’s ,20, and Therefore, considering these discrepancies in serum lipoprotein levels, and their possible influence on the progressive functional disability of the disease, the objective of the present study was to describe the lipid profile of patients with MS and its relationship with nutritional habits, as well as the possible improvement of functional capacity and the involvement of different lipoproteins in these changes, after EGCG and coconut oil intake.Study design and interventionA pilot, descriptive, quantitative, and experimental study was conducted. The clinical trial ID was NCT03740295. The population sample was obtained from the main state-wide MS associations who were previously informed about the study. The selection criteria and consort diagram flow for the experimental sample, consisting of 51 patients with MS randomly distributed into a control group (CG) and an intervention group (IG), is described in the study by Benlloch et al. , conductSerum extraction procedure and lipid profilingBefore and after the intervention, fasting blood samples were obtained from all participants. These samples were centrifuged to separate serum from plasma, and the TC, LDL, HDL, Apo A1, and TG levels were measured from the plasma. All the analytes were calculated by commercially available assays (Beckman Coulter) in an automated biochemistry analyzer .Nutritional analysisNutritional Dietary AnamnesisEach patient in the population sample underwent an anamnesis to obtain information regarding certain individual aspects such as personal and family history, usual medical treatments, comorbidities, food allergies or intolerances, and physical activity habits and the Food Frequency Questionnaire (FFQ) was employed . LikewisEating habits and dietary analysis®—Consultation Management Program” EasyDiet® (www.easydiet.es) [Likewise, an assessment of the patients’ diet was carried out through the nutritional-dietary software “EasyDietdiet.es) . The datStatistical analysisLipid profile values were available for the same 46 patients in which inflammatory markers were previously analyzed . Only ont test for independent samples or the Mann–Whitney U test, for normal or non-normal distributed data, respectively. Comparisons of means within groups to determine effect of the treatment were made with Student’s t test for dependent samples or the Wilcoxon test, for normal or non-normal distributed data, respectively.Statistical analysis was performed with the program SPSS v.23 . Categorical variables were reported as proportions and compared with chi-square test or Fisher’s exact test, when at least 25% of cells had expected counts less than 5. Continuous variables were reported as mean and standard deviation and normality was assessed by Shapiro–Wilk test. Comparison of means between groups was made with Student’s In addition to the mean comparisons of lipid profile values, linear regression models were constructed for a better understanding of the associations of the lipid profile with functionality and inflammatory process prior the intervention. No more than 7 predictor variables were used in each model, the combination of variables was chosen to avoid collinearity, with a variance inflation factor (VIF) < 2. To assume the independence of errors, values between 1.5 and 2.5 in the Durbin–Watson test were accepted. For each functional , and two-minute walk test (2MWT)), anthropometric (muscle mass and fat mass), or dietary  parameter lipid profile molecules were used as predictor variables, together with possible confounders such as: sex, age, BMI, and EDSS. For the EDSS model, only sex, age, and BMI were placed as confounders. For anthropometric variables (fat mass and muscle mass), BMI was excluded from the analysis. Finally, associations after the intervention were assessed by partial Pearson correlations controlling for age and sex in both the control and intervention groups. The level of significance in all analyses was α < 0.05.Ethical ConsiderationsThe study was developed in accordance with the Declaration of Helsinki , once th2. The general levels of TC, TG, and LDL in the patients of the sample were 217 mg/dL, 107 mg/dL, and 132 mg/dL, respectively. The HDL and Apo A1 levels were 83 mg/dL and 175 mg/dL, respectively. No statistically significant differences were found between men and women in any of the parameters described, except for Apo A1 concentrations, which were 13 mg/dL higher in women. No significant differences were found in the proportions of MS type between men and women, with a higher proportion of relapsing–remitting MS for both sexes.Anthropometric and lipid profile values analyzed by sex are shown in p < 0.001) due to the effect of BMI. However, when each type of lipoprotein was explored, LDL showed a significant positive association , indicating that high levels of this molecule are associated with a higher percentage of fat mass, as well as a significant effect of sex and BMI, with a total variability of 76.4% explained in this model . For muscle mass, a relationship with LDL was also observed, which in this case is negative , indicating that for a high percentage of muscle mass there are lower levels of LDL, with a significant effect of sex and level of disability, adding up to a total of 52.8% of the variability explained in this model . In addition, muscle mass showed a significant negative relationship with TC levels , with a significant effect of sex .Regression analysis for lean mass and muscle mass with lipid profile parameters showed a significant effect only for LDL levels on both parameters; all coefficients and significance values can be seen in The EDSS did not show any association with the lipid profile . HoweverAfter analyzing the dietary records of the 45 patients with MS who participated in the current study, it was seen that they consumed a high amount of foods rich in fats, among which the high consumption of simple sugars , processed meats, and snacks stands out, as shown in Since the intake of foods high in fat is a source of cholesterol and saturated fats, regression analysis was performed for the lipid profile and intake of total lipids, saturated fats, and cholesterol. No relationship was found between these parameters, nor was there a significant effect of any of the confounding variables . The coefficients and statistical significance can be seen in n = 20) and IG (n = 25) presented the characteristics and initial levels indicated in p = 0.034).Participants in the CG  in the CG, and from 220.9 ± 35.96 mg/dL to 232.5 ± 41.89 mg/dL  in the IG. HDL increased only in the CG, from 76.6 ± 15.2 mg/dL to 86 ± 14.2 mg/dL ; in the IG, HDL levels remained constant around with 84.8 ± 18.1 mg/dL and 87.9 ± 16.2 mg/dL in the pre and post measurements, respectively. TG levels did not show significant changes in any of the groups ). However, when lipid parameters after treatment were analyzed by two independent sample means, only TG levels showed significant differences, with lower TG levels in the IG than in CG . The other parameters, LDL and Apo A1, did not show significant differences in any of the groups.Lipid profile parameters showed some differences after treatment . TC leveThe lipid profile showed relationships with functionality parameters only in the IG . EDSS shSimilarly, body fat showed different associations with the lipid profile depending on the group B: in thep = 0.002) and Apo A1  only in the IG  A and then the IG C, indicaLipid control is common in clinical practice as it is a risk factor for cardiovascular diseases, although it is also beginning to be linked with neurodegeneration ,42. SpecIn our sample, we were able to determine a dyslipidemia characterized by high levels of TC and LDL: 217 mg/dL and 132 mg/dL, respectively. The values of Apo A1 and TG were within recommended limits: 175 mg/dL and around 100 mg/dL, respectively. Another aspect to highlight is the high levels of HDL, around 80 mg/dL. Metabolic alterations, including dyslipidemia, have already been suggested in this disease. Recent published data show that MS patients with a similar age range to the present sample (between 40 and 55 years) also had at-the-limit or above recommended TC and LDL levels ,45. The After the 4-month-intervention, it should be noted that the only significant difference between groups was due to a decrease in TG levels seen in the IG . This chIt should be noted that no changes in HDL levels were found in the IG, but significant increases were observed in the CG, which could be due to the effect of the healthy baseline diet received for 4 months (starting from slightly lower HDL levels than those found in the IG), coinciding with what was seen by Fellows et al., 2019, where HDL levels also increased after dietary intervention in a population with progressive MS . HoweverTo understand how our treatment achieves a paradigm shift in the lipid profile, possibly based on the new role of HDL and also related to the decrease in TG, we need to analyze the activity of both EGCG and the ketone bodies promoted by the intake of coconut oil. Regarding EGCG, it stands out that it upregulated autophagic lipolysis in adipocytes , which cThis is consistent with our results, where no changes in LDL concentrations occur after the intervention. Moreover, this lipolytic activity promotes a decrease in deposited TG and the synthesis of free fatty acids, which cannot only increase the production of ketone bodies in the liver but also promote the remyelination of neurons after crossing the BBB . It is aAt this point, it is interesting to note that the lipid profile influences inflammation and vice versa. Specifically, on the one hand, lipoproteins rich in TG and TG themselves are associated with an increase in the production of proinflammatory cytokines and the activation of monocytes  and, on As brilliantly established by Cho et al., 2020 regarding neuromyelitis optica spectrum disorder, serum levels of TG show a positive association with EDSS scores, with changes in HDL levels depending on the disease phase . TherefoAll of these hypotheses and results are shown in Despite these findings, our study has limitations. Being a pilot study, the population sample is small and a more representative population is required. In addition, it would be interesting to explore the change in HDL composition, quantifying the different subtypes. Finally, a certain degree of inaccuracy cannot be ruled out in food diaries that are self-reported by the patient and the possibility of this bias should be considered. Therefore, it is necessary to further investigate lipid metabolism and its role in the progression and prognosis of MS.As a conclusion, patients with MS show dyslipidemia characterized by elevated values of TC, LDL, and HDL, while TG and Apo A1 are within normal range. These lipid levels are not associated with nutritional habits. In addition, EGCG and coconut oil improve the functional capacity of patients with MS. This improvement could be due to the treatment’s impact on reducing blood TG levels; thus, TG in particular could be especially relevant as a prognostic biomarker of the disease.


Despite a recent global decrease in suicide rates, death by suicide has increased in the United States. It is therefore imperative to identify the risk factors associated with suicide attempts to combat this growing epidemic. In this study, we aim to identify potential risk factors of suicide attempt using geospatial features in an Artificial intelligence framework.We use iterative Random Forest, an explainable artificial intelligence method, to predict suicide attempts using data from the Million Veteran Program. This cohort incorporated 405,540 patients with 391,409 controls and 14,131 attempts. Our predictive model incorporates multiple climatic features at ZIP-code-level geospatial resolution. We additionally consider demographic features from the American Community Survey as well as the number of firearms and alcohol vendors per 10,000 people to assess the contributions of proximal environment, access to means, and restraint decrease to suicide attempts. In total 1,784 features were included in the predictive model.Our results show that geographic areas with higher concentrations of married males living with spouses are predictive of lower rates of suicide attempts, whereas geographic areas where males are more likely to live alone and to rent housing are predictive of higher rates of suicide attempts. We also identified climatic features that were associated with suicide attempt risk by age group. Additionally, we observed that firearms and alcohol vendors were associated with increased risk for suicide attempts irrespective of the age group examined, but that their effects were small in comparison to the top features.Taken together, our findings highlight the importance of social determinants and environmental factors in understanding suicide risk among veterans. AccordiOther relevant proximal factors include access to means and substance misuse , 18. ForThe objective of the present research was to conduct an analysis of climatic and socio-demographic factors that are associated with increased risk for suicide attempts among U.S. veterans using an explainable artificial intelligence (X-AI) model. We were also interested in the relationship of the number of firearms and alcohol vendors per 10,000 people as proxies for access to means and decreased restraint, which were also included in the final model. Together, we identify several novel factors at zip code-level resolution that impact individual-level risk for attempting suicide.2.2.1.2.1.1.The cohort and suicide attempt phenotype used in this study were initially described in Kimbrel et al. . A total2.1.2.The climatic features included two groups: static measurements and monthly measurements. Monthly measurements included longitudinal features such as monthly average precipitation and maximum temperature. There were 12 distinct measurement types recorded each month, totaling 144 features. The 30 static features included features such as elevation and percent urban cover. This led to a total of 174 climatic and weather-related features, mapped to 33,144 ZIP codes across the U.S.  Supplem.2.1.3.The socio-demographic features were collected from the 2019 American Community Survey, produced by the United States Census Bureau . These 12.1.4.From the Historical Business Database  we extra2.2.In this study, we used iterative Random Forest (iRF) , 37, an High pairwise correlations among input features can have a negative impact on the explainability of iRF models. This occurs because when one feature is strongly correlated with others, its importance is divided across the correlated features. As a result, the overall importance of these features in predicting suicide attempts decreases. To identify highly correlated features, we calculated Pearson’s correlation coefficients between all 1,784 features. Through this analysis, we identified feature groups with correlation coefficients equal to or greater than 0.90 in absolute value, and therefore considered as highly correlated. From each feature group of strongly correlated features we selected a representative feature to be included in the iRF model. Thus, this led to the removal of 248 features, reducing the total number of features to 1,536 . Please 2.3.true positives/true positives + false positives and recall as true positives/true positives + false negatives. Each random forest in the iRF model includes 1,000 trees with a leaf node size of 1,000 patients. We set the number of iRF iterations to five to rank the importance of each feature in predicting suicide attempt. In addition to ranking input features by importance, we next identified the feature-level explainability of our model by determining whether each feature was predictive for or against suicide attempt. To estimate if a feature predicts suicide attempt or controls, the result of each split was averaged using the given feature and mapping those to a linear effect. This provided both the feature effect in the slope of the line and an R2 with how closely that related to each of the set of splits. If the slope of the line was positive, then the feature effect size direction was positive, i.e., the value and the feature were positively correlated. If the slope of the line was negative, then the feature effect size direction was negative.To obtain accuracy scores for the iRF model, 5-fold cross-validation was used. The cross-validation technique employed in this study was group shuffle split. In this methodology, the dataset was randomly divided into 80% for training and 20% for testing, while incorporating a grouping factor to ensure that samples from the same group were not utilized for both training and testing in the same run. By employing patient zip codes as a grouping factor, we aimed to mitigate geospatial bias in our analysis. To further enhance the robustness of our analysis, we replicated this process five times, resulting in a total of five accuracy estimations for the models. Prediction accuracy was calculated using the average Area Under the Precision Recall curve (AUPRC), where precision was defined as 2.4.Model selection was based on research interest and accuracy gain. Initially, we aimed to address three questions: predicting suicide attempts based on climatic features, demographic features, and access to firearms and alcohol. We used iRF models with k-fold cross-validation for each feature group, and later employed all 1,536 features together to identify the most important predictors for suicide attempts across all feature groups in patients above or equal to 60 years and below 60 years. We then combined the top 20 most important features from the model with all features with alcohol and firearm vendor data per 10,000 residents. The reduced models showed better accuracy than using all features, as indicated by the area under the precision-recall curves (AUPRC) across 5 data splits , 3. Usin2.5.To show how features or groups of features interconnect each other, we applied iRF-Leave One Out Prediction (iRF-LOOP), which is an extension of the iRF model . In this3.3.1.firearms and alcohol vendors per 10,000 residents  at home were associated with reduced risk , and wind speed late spring through summer (which represents wind speed in May through September) were associated with decreased risk, whereas wind speed in autumn through spring (which represents wind speed for the rest of the year) and temperature in late autumn through winter  were associated with increased risk  and Water vapor in mid spring through mid autumn (which represents water vapor from April through October and minimum temperature in May) were associated with reduced risk  between features. Edges are weighted by their normalized feature importance value and the arrow direction shows what feature is predicting another one. Several features related to geographic mobility, population density, low gross rent with cash, and high home value were observed. Further, the subnetwork shows associations among features including number of widowed females, and several European ancestries. Regarding climate features, we observed temperature in late autumn through winter and temperature in late spring through summer, which represents average temperature from May to August, maximum temperature in June, and minimum temperature from June to September.It is well known that alcohol abuse increases suicide risk . Thus, w4.Most prior suicide prevention studies have focused on a relatively small number of features. Moreover, most have typically relied on individual-level information only  or aggregated data for a given geographical area with low geospatial resolution. In the present study we showed that ZIP code-level data can improve prediction in individuals with a history of suicide attempt greater than random chance, supporting the role of the surrounding environment as proximal/precipitating factors that influence the propensity for an individual to attempt suicide.Our selection of the iRF model was based on its suitability for our research objectives. In particular, its capacity to efficiently handle large, high-dimensional datasets was crucial for the present study. Furthermore, the model’s feature weighting technique helps to address data overfitting by prioritizing the most informative features while removing the non-relevant ones. This method also enhances the interpretability of the results by generating a ranked list of the most influential features. Lastly, the use of decision trees allowed for the estimation of the directionality of the prediction, enabling a deeper interpretation of the environmental factors contributing to suicidal attempts. Thus, by using this X-AI we were able to screen more than a 1,700 demographic and climatic features to obtain several that appear to potentially protect or increase veterans’ propensity for attempting suicide.An individual’s environment can have profound effects on their psychological state and subsequent risk of suicide. In fact, it has been well documented that socioeconomic and demographic factors such as poverty, education and population density are related to suicide risk . In our Interestingly, the top features can be classified into nine main groups namely: emotional support, housing, ancestry, commuting and mobility, climate, decreased restraint, access to means, cognitive difficulties and access to healthcare , 5. In tRegarding the commuting and mobility group, commuting time has been associated with depression in a dose responsive manner . Our resNative Hawaiian and other pacific islander females between ages 45 to 54 was associated with suicide attempts. These results are consistent with Ji et al.  revealed a prevailing pattern of increased suicide deaths occurring during the spring and early summer months . A subseSeveral mood disorders related to suicidal behavior such as depression, bipolar and seasonal affective disorder (SAD) also show seasonality patterns . A plausIn the present study, we found that solar radiation in summer predicted suicide attempt in patients younger than 60 years. Consistent with this result, a study found that intentional drug overdose deaths in the United States were higher with longer day lengths, which correlates with months with higher solar radiation . ConversHigh temperatures are linked to suicide rates , and theNumerous studies have explored the relationship between water vapor (humidity), rainfall, and suicide risk , 15. HowLimited research also exists on the relationship between wind speed and suicide risk. Some studies yield contradictory or inconclusive results . In our Although terrain elevation is not inherently a climatic feature, we classified it as such due to its association with various climatic factors that undergo changes alongside it. Our study revealed a positive association between altitude and suicide attempts, aligning with a systematic review that showed a similar trend in 17 out of 19 analyzed studies , 71. HypAnother goal of the present work was to explore decreased restraint and access to means for suicide attempts using the number of alcohol and firearms vendors per 10,000 residents as proxies. Importantly, regardless of age, we observed that living in areas with more alcohol and firearm vendors was associated with increased risk for attempting suicide . These results provide additional evidence regarding the importance of access to means as one of several risk factors for suicide, especially in the U.S., where a larger proportion of suicides are committed using firearms in comparison to other high-income countries where only 5% use them , 73, 74.In this study we demonstrated the use of X-AI to explore the impact of more than 1,700 demographic and climatic features on suicide attempt risk with high geospatial resolution. This research provides additional evidence for the role of several demographic and climatic features in suicide attempts and demonstrates the utility of using geospatial features from the area  in which patients live within an X-AI framework to improve suicide risk prediction. By focusing solely on sociodemographic and environmental factors, we aimed to determine their distinct influence on suicide risk, regardless of clinical or psychiatric factors. We also aimed to investigate their influence from a public health standpoint. Analyzing these factors can provide valuable insights for developing effective interventions and policies to decrease suicide risk. By identifying the key factors that contribute to suicide risk, researchers and policymakers can prioritize interventions that specifically target these factors. For example, understanding the localized variations in risk factors can help target interventions to specific regions or communities. Suicide prevention measures can be tailored based on the unique sociodemographic and environmental characteristics of different areas, allowing for more effective and efficient allocation of resources. Interventions could include promoting social support networks, providing tailored mental health services, and implementing community-based programs that address the unique needs and challenges of vulnerable populations. Overall, the findings contribute valuable insights to suicide prevention measures by highlighting the role of sociodemographic and environmental factors and emphasizing the need for a holistic, geospatially-informed approach. By integrating these findings into policies and interventions, it is possible to develop more effective strategies for preventing suicide and promoting mental well-being in at-risk populations.4.1.The present study had a number of limitations that should be considered when interpreting these findings. First, we utilized a cross-sectional design, and a lifetime suicide attempts variable. Thus, additional work is still needed to examine the degree to which the features identified in the present study might be predictive of future suicide attempts. Second, our findings regarding alcohol and firearms vendors should be interpreted cautiously, as there are several other features  that couSuicide attempt data from individual patients is not readily available because this data is considered Protected Health Information by the U.S. Department of Veterans Affairs. All climate data and socio-demographics can be made available upon request.The studies involving human participants were reviewed and approved by VA Central Institutional Review Board (cIRB# 18-11). The patients/participants provided their written informed consent to participate in this study.MP, JL, AW, KS, MG, NK, and DJ: conceptualization. MP, JL, AC, JR, and DJ: methodology. MP: validation, investigation, and visualization. MP and AW: formal analysis. MP, AW, and JS: data curation. MP, KS, JS, MG, JP, and NK: data interpretation. MP, JL, and AW: writing of the original draft. MP, JL, KS, JS, MG, BM, DO, JB, NK, and DJ: writing—review and editing. DJ: funding. NK and DJ: supervision. All authors contributed to the article and approved the submitted version.This research is based on data from the Million Veteran Program, Office of Research and Development (ORD), Veterans Health Administration (VHA), and was supported by award #I01CX001729 from the Clinical Science Research and Development (CSR&D) Service of VHA ORD. This work was also supported in part by the joint U.S. Department of Veterans Affairs and US Department of Energy MVP CHAMPION program.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.http://energy.gov/downloads/doe-public-access-plan, last accessed September 16, 2020).This publication does not represent the views of the Department of Veteran Affairs or the United States Government. This manuscript has been co-authored by UT-Battelle, LLC under contract no. DE-AC05-00OR22725 with the U.S. Department of Energy. The United States Government retains and the publisher, by accepting the article for publication, acknowledges that the United States Government retains a nonexclusive, paid-up, irrevocable, world-wide license to publish or reproduce the published form of this manuscript, or allow others to do so, for United States Government purposes. The Department of Energy will provide public access to these results of federally sponsored research in accordance with the DOE Public Access Plan (