Mycobacterium tuberculosis, leading to the discovery of a new derivative of a known drug. Work on penicillin during and after WWII lead to the development of associated salts/esters and a search for new natural product antibiotics. This led initially to no new, marketable compounds, but concluded with the serendipitous discovery of fusidic acid, an antibiotic used to treat infections by Staphylococcus aureus, in 1960. The discovery process included contemporary approaches such as open innovation; targeting specific pathogens and/or specific organs in the patient; examining the effects of antimicrobial compounds on bacterial virulence as well as on antibiotic-resistant variants, and searching for antibiotic producers among microorganisms not previously well explored. These activities were promoted by the collaboration with a renowned Danish clinical microbiologist, K. A. Jensen, as well as company expertise in fermentation technologies, chemical synthesis and purification of bioactive compounds from organic materials.The 1940s and 1950s witnessed a diverse search for not just natural product antibiotics but also for synthetic and semisynthetic compounds. This review revisits this epoch, using the research by a Danish pharmaceutical company, LEO Pharma, as an example. LEO adopted a strategy searching for synthetic antibiotics toward specific bacterial pathogens, in particular  This review covers the 1940s and 1950s that witnessed the golden age of discoveries of naturally occurring antibiotics , with thThe discovery approach for naturally occurring antibiotics was simple and consisted of screening primarily soil microorganisms on a range of agar media for antagonism toward selected target organisms: the Waksman platform , althougStaphylococcus aureus while in other cases such as vancomycin resistance in enterococci it took several years  included more than 5000 sulfonamide derivatives by the end of WWII . After Wacterium . The intacterium . In contChemical synthesis of naturally occurring antibiotics turned, on the other hand, out to be too difficult or expensive in most cases with chloramphenicol as the prominent exception. Instead, there are examples early on of the development of derivatives of natural fermentation products (semisynthetic antibiotics) such as dihydrostreptomycin and tetracycline. Overall, the value of semi-synthetic antibiotics was recognized from the work on β-lactams at the end of the 1950s.I illustrate the work done on antibiotics in the 1940s and 1950s by examining one particular example: the research done by the small Danish company LEO Pharma. This company manufactured at that time several different drugs, many of them belonging to other categories than antibiotics. The company was active in research and development of natural product, synthetic and semisynthetic antibiotics frequently in close collaboration with an internationally renowned medical microbiologist (K. A. Jensen) with strategies depending to a large degree on target species and pharmaceutic issues. K. A. Jensen was involved in the first result from the empirical search for natural product antibiotics: the re-isolation of penicillin during WWII. Subsequent screening for naturally-occurring compounds with its failures and ultimate success was more a company activity that only initially involved another external collaborator in a supportive role. The work by LEO focused also on challenges such as antibiotic resistance and targeting virulence properties of bacterial targets, a potentially important strategy to counteract selection for resistance. Finally, it is of interest how LEO supported the innovation pipeline, by applying the open-source approach and searching for new natural product antibiotics among poorly investigated microorganisms.Streptomyces) due to it has been used in a number of sources I quote. The primary source material on antibiotic research from 1940 to 1960 is in the archive of LEO Pharma Historical Archives and Museum. My manual search of this archive, across numerous visits, was essential to the completion of this review. This archive supplied meeting memos and summaries, letters and research reports, timesheets for chemists 1946–1959, two hand-written laboratory notebooks for the period from 1951 to 1956, orders for target strains for screenings and lists outlining the contemporary organization of the company into research units. Finally, I interviewed a researcher who was involved in the discovery of fusidic acid at LEO.The material for this review comes from different sources. The scientific literature offered substantial information, including quantitative data for discovery rates for actinomycetes antibiotics 1940–1965 obtained from 2. Examples include anesthetics , diuretics  and hormones  in addition to antibiotics. The company also worked on several smaller development projects and product categories. The majority of drugs were synthetic compounds whereas the hormones constituted natural compounds extracted from animal tissues or human urine. The know-how in natural product chemistry obtained by working with hormones proved important concerning subsequent purification of natural product antibiotics.The purchase of a pharmacy store in 1908 marks the beginning of the company. In 1909, LEO started production of a yoghurt preparation, and marketed an aspirin product soon thereafter. Then, in 1917 was launched the first Danish export drug, Digisolvin, a standardized Digitalis-based product. Hormones became a part of the product portfolio from the 1920s . During 3. From 1946 to 1959, around 50% of research time was devoted to antimicrobial compounds  l trials , 2016. A6. Developing new antimicrobial compounds toward this disease constituted an important activity at the company until the end of the 1950s 11.The situation after WWII with tuberculosis on the rise again in Europe, contributed to the urgency of the efforts as was also the case for other pharmaceutical companies such as Bayer in Germany 6. Develohe 1950s , 37. The12. Tuberculosis requires a long period of antibiotic treatment, which increases the probability of selection of resistant variants. This aspect had the attention of the scientific community already in the late 1940s  was developed with a less irritating effect on patients than PAS and therefore suitable for the long duration of chemotherapeutic treatment of tuberculosis  whereas others were new or with unknown antimicrobial activities . The compounds were, in collaboration with K. A. Jensen, tested toward a panel of bacterial targets including anaerobic bacteria, enterococci, E. coli, pneumococci, streptococci and “proteus bacteria.” Some of the results appeared promising at first, but no lead compounds were found20.Chemical synthesis of antimicrobial compounds, many generics, targeting other infectious diseases than tuberculosis took place during the 1940s and 1950s. Three anti-syphilitic agents were synthesized, and one of these was on the market from 1941 to 1953 18. The a21. The archives do not contain information on the objectives nor the outcome of this research. Most likely, it was inspired, by the launch by Pfizer in 1953 of tetracycline, a semisynthetic product obtained by cleaving the carbon-chlorine bond of the natural compound chlortetracycline  asking K. A. Jensen in 1942 whether he would be interested in extending their collaboration to include this topic. Jensen isolated the same year a penicillin-producing mold from the air in his laboratory, similar to Fleming’s discovery in 1928. LEO collaborated with K. A. Jensen on developing the manufacturing process that went on in secrecy due to the German occupation of Denmark during WWII . The comacturing . Produci23. One outcome was Leocillin  with a high lung affinity that found application as a veterinary product  is of considerable importance to investigators who apply for process or product patents”  … that the substances one finds using a given screening method are not necessarily most effective against the target of the screen”  and three (two of them erythromycin and vancomycin) antibiotics on the market and it is not surprising that LEOs limited efforts in the first half of the 1950s led to nothing. If the screening at LEO is compared with the work by two other small research units at the time, the ETH laboratory in Switzerland and the laboratory run by Salman Waksman at Rutgers University, the negative outcomes for LEO and the ETH were similar but opposed to the Waksman laboratory. The latter, however, initiated their work much earlier resulting in discoveries of several actinomycetes antibiotics, four of which found practical applications: Actinomycin (cancer), streptomycin (tuberculosis), neomycin , and candicidin . In all three cases actinomycetes antibiotics constituted a major focus but most of the low hanging fruits were gone by the time LEO and ETH initiated their work .in vivo inhibitory activity toward Mycobacterium tuberculosis using guinea pigs as an experimental model. The compound had not been isolated and therefore its chemical composition and structure was unknown. There were indications that it did not resemble known antibiotics used chemotherapeutically at the time. The company would aim at reproducing these findings and in case of an affirmative result, would conduct further investigations to explore possibilities for manufacturing the antibiotic in quantities for the use of chemotherapeutic treatment of human tuberculosis patients. In case of a positive outcome, the doctor was interested in LEO producing the compound on basis of a license36. Most likely, results did not support this prospect and no other documents in the archive refer to this case.It is of interest that regarding natural product antibiotics LEO looked during the early 1950s for opportunities when possible. In one example, the company participated in an investigation that was a forerunner to the open innovation concept; a concept that recently had become popular among pharmaceutical companies including LEO Pharma . A confi37.Another example of the search for the potential chance discovery was the testing of an antibiotic-producing culture accidentally obtained from the lab environment, however again without resultFusidium coccineum (now Acremonium fusidioides) strain, an organism they had not heard about before. They decided to “very unscientifically to buy it – just because of the similarity of the name with Fusarium” – organisms they already used (quotation from S. aureus (Success finally arrived with the discovery of fusidic acid in 1960, a serendipitous result of the work on semisynthetic β-lactams around this time . This ap. aureus .Streptomyces or molds but no further R&D appear to have taken place (It is instructive that LEO’s first real successful hit came when they examined the antibiotic potential of a genus of molds not tested previously, although related compounds, helvolic acid and cephalosporin P1 produced by other genera of molds were discovered much earlier, in 1943 and 1951 . Searchien place . The comThis study explores the variety of strategies employed during the 1940s and 1950s for finding new natural, semisynthetic and synthetic antibiotic compounds. It is of interest, that LEO was a pharmaceutical company of relatively small size. Further, the owner from 1940 and until 1986, Knud Abildgaard aimed at rapidly securing marketable results from research. This offered less opportunity for larger strategic efforts, as opposed to numerous smaller projects. Although Abildgaard did not have a pharmaceutical or chemical background, he had, however, obtained an extensive amount of pharmaceutical knowledge . Taken tResearch on synthetic antibiotics and salts and esters of penicillin benefited from the collaboration with K. A. Jensen as he tested antimicrobial activities, organized clinical trials and gave essential input as a consultant. He and other consultants also participated in regular meetings with the management of LEO. These meetings concluded by dinners that also served as incubators for professional interactions and even friendship and thereby fulfilling a role as a basis of knowledge and insights that served LEO well in their research . The impInterestingly, the work on natural product antibiotics involved a collaborator, H. A. Jensen serving only a supportive role. This situation was not too different from the research that led to the discovery of chloramphenicol. “In 1943, no doubt alert to the work that Waksman was doing at Rutgers, but not yet wishing to commit company resources to an uncertain field of study, Parke, Davis and Co. in Detroit provided the funds for Paul Burkholder, Eaton professor of botany at Yale to screen samples of soil for micro-organisms producing substances with antibiotic activity” , p. 219.M. tuberculosis, and the search for compounds with a localized activity, e.g., an improved lung affinity. Further, the search included compounds with effect on antibiotic-resistant targets and/or on virulence rather than survival. In addition, the discovery of fusidic acid was a reward for testing microorganisms not previously well explored. Finally, there was an example of open innovation, a management mechanism that supports discoveries by searching outside the search parameters set by the company. All of these approaches are also in one way or another in play in the antibiotic discovery process today (In overall, the work by LEO came some way to meet the challenges at the time for finding new antibiotics. These included a need to be more precise in targeting the pathogens. This was the objective with the screening for antimicrobial compounds toward ss today .JL designed the study, performed the research, and wrote the manuscript.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Nipa palms, a plant species in mangrove forest, are valuable and beneficial for the local community's economy as well as the conservation in the southern region of Thailand. This study aimed to investigate the use of nipa palms in Khanap Nak and focused on the type of products made from nipa palms and the yield of this plant through focus group discussion and interview with the farmers maintaining nipa palm forest. The results suggested that nipa palms in Khanap Nak can yield for 5–100 years. Apart from the benefits to the community in terms of sustainability, as it prevents erosion, it provides sources for the production of food  and raw materials , which can generate income to community members. It was found that most of the communities earn 90–130 USD/day from palm sugar production. The quantity of nipa palm products varies in each season, resulting in different prices. In this regard, they can produce high quantity of products made from sap from January to March. However, the production of different types of sugar requires local knowledge and wisdom to ensure good quality. Nipa palm production is the sustainable way to utilize mangrove forest resources, leading to effective conservation and good life quality. Regarding problems and difficulties in farming, it was found that natural disaster is a major threat, such as drought, excessive amount of salt or freshwater in certain periods, and insect pests. Nypa fruticans Wurmb.) is the only species able to adapt to mangrove forest. This trunkless palm dominates wetlands commonly flooded by brackish water [Thailand is located in the tropics, which has a diversity of mangrove forest . Varioussh water –7. This sh water .Located in Pak Phanang Basin, Khanap Nak is the largest source of nipa palms in Thailand . Nipa paSustainable utilization of nipa palms can make environmental, social, and economical impacts . The sapCenturies ago, coastal communities in Pak Phanang Basin have traditionally been utilizing nipa palms, but the empirical evidence of production and consumption of the processed products from nipa palms is still limited. Similarly, the value of nipa palms on environmental, social, and economical aspects is the contribution to sustainable utilization of forest products. In connection with this, the present study aimed to identify a variety of nipa palms' sap products utilized by local communities in Pak Phanang Basin. It was expected to contribute to the literature regarding the role of nipa palms in these communities, income generation, and its environmental value.This research was conducted in Khanap Nak, located in Pak Phanang Basin, Nakhon Si Thammarat, Thailand, which is the largest source of nipa palms in Thailand , 9. The Key Informant Interview. The selected key informants for this research were related to nipa palms, including the community representatives who were directly and indirectly involved in the production and distribution of nipa palm products. They were selected through snowball sampling based on the characteristic suitability of the subjects and the requirements to obtain complete, accurate, and consistent data. There were a total of 12 informants participating.Brainstorming and Focus Group Discussion. This allowed people with nipa palm plantation to participate in providing data regarding values, products, and lifestyles related to nipa palms.Interview with 100 Households (of 389) of Nipa Palm Producers from 10 Villages. The sample size of this study is more than 25% of the population for the small population [pulation . The strSurvey and Direct Observations of Nipa Palm Forest and Farms in Khanap Nak. The productivity of each farm of people was surveyed to determine the yields.Data were obtained from two sources: primary and secondary. The primary data were collected from January to October 2019, while the secondary data were gathered or collected from various sources, such as documents issued by the Department of Marine and Coastal Resources, journal articles, official records, and project reports. The values, lifestyles, and importance attached to nipa palms as well as its role in income generation were investigated. The process involved the following methods:This study specified the sustainability criteria to assess the opportunities of nipa palm product utilization from environmental, social, and economical sustainability aspects . The obtKhanap Nak has an area of approximately 38.8 square kilometers with 10 villages and a total population of 5,295 people. The community is a coastal community with a large amount of nipa palms. Therefore, the community's lifestyle is linked with nipa palms and people rely on making nipa palm products for primary and supplementary income. According to the findings of this study, the community is a home to 389 households relying on nipa palms for living. A survey indicated the palm forest area of 5.76 square kilometers in this community. The results from the farmer interview revealed the age of palm trees in this area is from 5 to 100 years. In addition, it was found from the interview and observation that nipa palms are crucial for the community in environmental, social, and economical aspects as presented in As pointed out earlier, nipa palms are a valuable resource in coastal areas of Thailand. Local communities rely on this plant for both environmental and economic benefits . ParticuThe knowledge about nipa palm cultivation is acquired through inheritance and has been passed on from generation to generation by the villagers of Khanap Nak. Since ancient time, the villagers have followed traditional methods for cultivating nipa palms and producing nipa palm sugar. The cultivation goes through a series of methods and processes and involves various equipment. The extraction also involves traditional techniques as shown in Every part of a nipa palm can be used for a variety of purposes. As cooking ingredients, nipa palms were found to be beneficial to people in the communities where the study was conducted. In Khanap Nak, young flowers were used to make food, especially by boiling and then dipping in chili paste. Similarly, fruits aged four months were pickled and cooked as a vegetable in curry. Fruits aged 5–8 months were used to make dessert, while sap from nipa palm flowers were used to make sugar. In addition, the sap can be used to make nipa palm sugar, powdered sugar, molasses, and vinegar. Phetrit et al. [In an interview, a farmer supporting the health value of nipa palms said the following.I have been eating nipa palm sugar since I was little. Also, my ancestors regularly consumed nipa palm sugar and no one has had diabetes, everyone is healthy.” .“Cotylelobium lanceolatum Craib), sliced into small flakes, is added into the bamboo cylinder in the extraction process. It contains polyphenols with acerbic taste, which act as antimicrobial and antioxidative agents [The preparation processes for each type of nipa palm syrup are summarized in e agents , 16, 17.Each type of sap requires different preparation methods and equipment based on community knowledge, which has been transferred from generation to generation. Each step requires experience and expertise, such as the insufficient addition of kiam wood flakes can cause sap to rot, while the excessive addition can cause too much bitter taste and ultimately affect the taste of the processed sap. Nipa palms' sap products can then be distributed and consumed in the household.In addition to using nipa palms for consumption, it was revealed that the residents in Khanap Nak use nipa palms to make various products; for example, water containers made of leaves, ropes for thatch sewing, and potholders made of soft branches. In some locations, fresh nipa palm tops aged two months are used to roll cigarettes, while old leaves and branches are whittled to make sticks of 1–1.20 meters length to hold thatches while sewing. The old leaves can be also woven into hats.Apart from consumption benefits and conservation value, nipa palms provide crucial benefits and values in improving the quality of life of local communities through generating income for many households. It was evident from the interview conducted with community members in Khanap Nak that nipa palm farming is a supplemental occupation for some households and a primary one for many households all year round. According to the interview, the households cultivating nipa palms as well as those making various products from the crop derived income directly from the venture. For instance, molasses can be sold at a price of 900–1,400 THB (30–46.7 USD) per bucket (25 kilograms), depending on the season, while granulated sugar at a price of 120–140 THB (30 USD) per kilogram and bottled syrup at a price of 60 THB (2 USD) per bottle. However, the income generated from nipa palm sugar is dependent on the quality of produced sugar influenced by factors, such as water quality, rainfall, and weeds.In terms of utilization, different products were found to be made from and of nipa palms in all 10 villages of Khanap Nak . Nipa paThe results from the survey and interview with 100 households engaged in nipa palm production showed that the dominant products from nipa palms were molasses (91 households or 80% of farmers), followed by vinegar at 12%, syrup and thatch at 3% each, and others, such as granulated sugar and cigarette paper, at 1%. Similarly, nipa palm molasses is one of the most important products invaluable for the economy of Bangladesh .According to The nipa palms' sap is a major ingredient in many products. As indicated, the farmers and community members use the traditional extraction techniques to produce these products as a way to generate income to support their living. Four main products obtained from the sap are discussed in detail as follows.The molasses production is the most popular product in Nakhon Si Thammarat. The interview with nipa palm farmers and survey results indicated that molasses is the main product for alcohol production . This prIt was observed that most of the molasses producers are elderly local people with an average age of 58 years, while nipa palm farmers had an average age of 85 years with extensive experience in farming on their own land and/or leased land. Therefore, this area has adequate labor force to engage on nipa palm farming. According to the interview, the nipa palm farmers 90.11% chose to make molasses because it is easy to sell the products to middlemen who buy directly from the farmers. In addition, the farmers indicated that molasses is in demand and can be sold throughout the year even though the price may insignificantly vary throughout the year. A farmer in Village No. 2 indicated the following in an interview.Molasses is easy to make, uncomplicated and, most importantly, it can be kept for years. I sometimes store 10–20 buckets before calling the merchant to come and pick it up at my house. I get a lump sum for it.” .“Nipa palm syrup is a popular product for consumption in Khanap Nak and other areas and for purchase by tourists visiting the region. However, few producers were found in this study. According to a survey, there were only 3.In an interview with a farmer in Village No.5, he spoke about nipa palm syrup production:Only few people make nipa palm syrup. Most of them don't. They've all gone on to make molasses because it's difficult and time-consuming to make nipa palm syrup. Most importantly, the cylinder and sap need to be kept clean.” .“It was found that Khanap Nak can produce nipa palm syrup at an average of 28.33 liters per day and a maximum of 60 liters per day. The spadices are currently required at an average of 126.67 spadices per household.In terms of labor availability, the community has 2.33 workers for production per household with an average age of 56 years and 17 years of experience in making nipa palm syrup.Regarding the choice of production, it was found that the farmers chose to produce nipa palm syrup due to market availability and continual purchase. The products can be sold in wholesale and retail sales at all time, resulting in continual production. However, the syrup production process is very detailed and the sap's purity or cleanliness needs to be maintained throughout the production process.The sugar flake production in Khanap Nak was found to be low. A survey of the communities showed that Village No. 5 has only 1 producer with an average production of 11.5 kilograms per day. 150 spadices are required for the production per household, and 5 workers engaged in production. As highlighted in Regarding the choice of production, the market demand for the product is the key factor. Furthermore, the produced sugar flakes can be easily sold in wholesale and retail sales. Despite the process requiring detailed procedures and patience, the diligence and need to sell different products from elsewhere prompt the farmers to produce sugar flakes for distribution. Thus, the sugar flake production was merged to the community's enterprises.As Hossain and Islam  reportedEast Asia is commonly known for nipa palm vinegar consumption . Nipa paIt was also revealed during the interview and survey that nipa palm vinegar is a healthy product and can be used to cook many types of local food. The nutrition and chemical components, such as sucrose, glucose, fructose, vitamins, and antioxidant activities, were found in nipa palm sap , 9, 20. Nipa palm thatch is another important product from nipa palm for making roofs and walls of dwellings , 22, 23.Regarding the choice of production, 50% of the farmers were found to use materials, such as nipa palm leaves. In dry season, the branches are trimmed and the trimmed leaves are used to produce thatch for sale and use in the community. In addition, the farmers indicated that thatch is easy to sell as the demand was high. After the production, they sell the remaining trimmed leaves to the merchants in the community. More importantly, they can benefit from low production costs due to the availability of nipa palm leaves in the area. Furthermore, ropes from khla trees grown in Khanap Nak are used in the production process. This study shows that leaves are less utilized than the sap. This finding is in contrast with the situation in Bohol, Philippines, where nipa palms are more utilized for thatch production than any other purposes [In the past, people smoked cigarettes using nipa palm leaflets for cigarette paper , 24, whiIn addition to product distribution, it was evident from the findings and discussion that nipa palm products have benefits and values in terms of improving the community's quality of life and generating income. The interview with the community members showed that the households engaging in nipa palm cultivation have the maximum daily income from molasses at 2,700–3,000 THB and the farmers can profit 1,200–2,000 THB per day maximum .+ in soil and the nipa palm growth control the sap production, it can be improved by both increasing the soil organic matter content and decreasing brackish water. In addition, the average maximum temperature for nipa palm is 32–35°C, and the optimal climate is subhumid to humid [Despite the production of various products from nipa palms, it is crucial to point out that nipa palm producers still face 4 main problems . Most ofto humid . The resIn an interview with a farmer in Village No. 1, he spoke the following.“This year is hot …no rain in the nipa palm for 8 months, so the flower inflorescence of nipa palm cannot grow and no sap for a long time. ” .Sampson et al.  admittedNipa palm forest is a source of food and raw materials for household instrument production and income generation for many local communities. The results of the study indicated the main products from nipa palm sap are sugar, vinegar, and alcohol, similar to the production in Bangladesh .Allow the community participation in maintaining nipa palm forest through management, planning, and utilization. Many communities have inherited the maintenance and utilization methods for nipa palm forest from generation to generation for a long period of time. Therefore, they adapt the principles of the sustainability management while benefiting from the resources provided by the forest.Maintain the good price for production and distribution of nipa palm products while supporting markets for the community to continually generate income.Develop nipa palm products to meet required standards with longer shelf life to increase product value.Therefore, the communities in the study area have maintained the nipa palm forest to prevent soil erosion and increased the forest size. The community members participate in the nipa palm forest management with oversight and support for sustainable utilization from the government agencies and implement the following management strategies:Khanap Nak is an area with high nipa palm cultivation and has utilized nipa palm to produce a variety of products for a long time. In particular, nipa palm syrup production is part of local knowledge and cultural heritage of the area. The leaves, branches, and flowers of nipa palms can be used for cooking, while the sap from the spadix can be used to make sugar and vinegar. Therefore, nipa palms and mangrove forest have diverse benefits and values in terms of environment, utilization, and income generation. The communities cultivating nipa palms have primary and supplemental occupations related to nipa palms. In conclusion, Khanap Nak is a model area for sustainable nipa palm utilization. Given the tropical area is optimal for nipa palm growth, the sustainable utilization would benefit the forest and improve life quality of people in the community.


This study allowed to realize that a great number of positive samples involved banned pesticides  but, at the same time, many positives cases were due to the exposure to commercially available products . Also, it's possible to identify the areas where domestic species are the most affected  and the areas where the wild animals are the mainly affected species .Nowadays the intentional poisoning of domestic and wild animals is a crime in the European Union (EU), but as in the past the poison is still used in rural areas of a number of European countries to kill animals that were considered harmful for human activities. From January 2014 up until October 2020, the Laboratory of Pharmacology and Toxicology of the Faculty of Veterinary Medicine (LFT-FMV) has done the analytical detection of poisoning substances in 503 samples of wildlife and domestic animals and pesticides residues were found in 239 of the samples analyzed. In this retrospective study, toxicology results from domestic species , wildlife species , and food baits, are presented. During this period the samples analyzed at the LFT-FMV, were received from all over the country. Analytical detections were performed via solvent extraction followed by thin layer chromatography. Molluscicides (47%,  Poisoning of wild and domestic animals is a problem in all the countries in the world that has been described by several authors from ancient times to nowadays –16.Acute pesticide poisonings of wildlife and domestic animals can occur due to accidents with substances approved for several uses, and due to deliberate and illegal attempt to poison animals even with baits, and by secondary poisonings .Pesticides or more correctly Biocides, group include a wide range of chemicals, with a great diversity of applications, including agriculture and domestic applications. However, pesticides are often implicated in acute intoxications, in particular insecticides like molluscicides , 17, 18.The occurrence of poisonings related to pesticides is influenced by many factors, such as the availability of the substances without restrictions to the acquisition, the possibility to buy them easily, the agricultural techniques, the cultural background and the knowledge and instruction of users, etc . In geneOne of the best examples is strychnine and aldicarb. These substances had been banned from the market for use in the agriculture and as a rodenticide (strychnine), in Portugal in 1988 and in the European Union in 2006 for strychnine , 22. ConIn this study, the incidence of pesticides detection from animals/carcasses, baits, and other materials sent to LFT-FMV for toxicological evaluations was analyzed, trying to provide a general overview on animal exposure to pesticides, in both domestic and wild species, for a period of the last 6 years.Portugal has an enormous geophysical diversity, which provide an important habitat for wildlife mostly on border with Spain on the North, Centre and South of the country, where lot of birds of prey can be found.In all the country but specially in the North and South of the country there several areas where animal protection programs, such as Life Rupis, are running.Since 2003 Portugal have a multidisciplinary program (PROGRAMA ANTIDOTO) to study and understand which and how toxic substances are used, in order to stop animals killing, particularly wild animals , but also domestic species that can be exposed to these substances and be intoxicated. This program recently has a great improvement by the creation of a network of necropsy centers and laboratories of Toxicology (LFT-FMV is one of these laboratories) that in conjugation with all the other entities involved try reduced animal poisoning in Portugal. So, in case of the identification of a poison bait and suspected poisoned animal, the competent/policies authority (SEPNA/GNR or PSP) must be informed and those materials should be sent to a laboratory service for necropsy and/or toxicological investigations in order to identify the potentially toxic substance present. If the poisoning events are confirmed, the competent authority has to be notified and legal prosecution may follow.The analyzed samples include biologics matrixes of the different species , and baits. These samples come from ANTIDOTO Program, but also from private requests for example from veterinary practitioners.Data analysis included the location of the poisoning, type of species and number of animals affected, and the pesticide(s) detected.Procedure for the extraction, purification, and detection of pesticides in samples from animal poisoning incidents have been developed and validated in LFT-FMV. The extraction was performed using specific solvents for each group, followed by separation and characterization by thin layer chromatography (TLC) in Merck silica gel G 60 F254.The detection of all substances was carried out by qualitative TLC, all the plates were spotted with the extract of the sample and the corresponding standards which allows to have always a positive control in each analysis. In addition to this, the measurement of the retention factor (Rf) of each compound was done and compared with the Rf of the sample , 24.For Strychnine the sample was minced, mixed in aqueous solution , and agitated with a magnetic stirrer during at least 3 h. After, the sample is decanted and a purification step is performed to remove coextractants with chloroform, after that the final step of the extraction procedure is a liquid-liquid extraction. The phase with chloroform was collected to a round bottom flask and evaporated until a dry extract was get. The final concentrated extract was spotted on a silica gel TLC with a mobile phase chloroform/methanol (9.5:0.5) (9.5:0.5) . After p9.5:0.5) .For Coumarinics the sample was also minced, but mixed in a solution of acetone and chloroform, and agitated with a magnetic stirrer during at least 3 h. After this period, the sample is decanted, and liquid-liquid extraction was made with water addition. Then the aqueous phase was discharged, and the organic phase was collected to a round bottom flask and evaporated until a dry extract was get. The final concentrated extract was spotted on a silica gel TLC with a mobile phase n-hexane/acetone (9:1) (ne (9:1) . After pne (9:1) .Finally, for Organophosphates/Carbamates and molluscicides, the minced sample was mixed with acetonitrile, and agitated with a magnetic stirrer during at least 3 h. After this period, the sample is decanted, and liquid-liquid extraction was made with n-hexane addition. Then the n-hexane phase was collected to a round bottom flask and evaporated until a dry extract was get. The final concentrated extract was spotted on a silica gel TLC with a mobile phase n-hexane/acetone (9:1) (ne (9:1) . After pne (9:1) .For all the groups, including anticoagulants , the detection limit is 2 μg.The analysis of pesticides in suspected samples was carried out, if possible, according to symptomatic and pathological evidence, data provided by veterinarians in the reports sent with the animals for necropsy or with the baits, in addition to epidemiological data .The great difficulty faced in these cases is to distinguish whether they are intentional or accidental poisonings.Data collected were organized in a database of Microsoft Office Excel® and descriptive analysis and graphics were developed and created also with Microsoft Office Excel®. Descriptive data are reported in absolute and percentual values .From the total number of 503 samples received between January 2014 and October 2020 with intoxication suspicion by pesticides the requests come from ANTIDOTO program  and from private entities .In the As can be observed in Concerning baits, most of the samples received over the years came alone and not associated with a specific animal species. But from the 69 positive cases of baits, 15 are associated with foxes, 6 with dogs and finally 2 with cats, for all the others no relationship could be established.n = 73). In the other group wild animals are those mainly affected, and samples comes from the districts of Faro, Castelo Branco and Bragança . These results are showed in the Portugal is divided into three main regions, North, Center, and South that includes 18 districts. Data obtained on this study allows the distribution of the results in two major groups. In the first one samples for pesticide detection comes from the districts of Setubal and Lisbon and includes the group where the most affected species are the domestic animals  and Carbamates  were found to be the first category of pesticides detected in samples of suspicious intoxications, in both domestic and in wild animals, followed by rodenticides, with first and second generation represented , by Strychnine  and in the end by Organophosphates . This total distribution by group of pesticide can be observed in the Figures 8–10, respectively.Molluscicides , cats in lower number  and a small amount of other species  .The most affected domestic animal species  are mainn = 20), red foxes , vultures , eagles , rooks , and a small amount of other species .Concerning wild species , the mosResults obtained in this study show that Molluscicides and Carbamates intoxication are still one of the major causes of poisoning in domestic and wild species in Portugal following the pattern previously reported by our team , 28.Portugal has favorable climatic conditions for snails and slugs. Baits are used in domestic and commercial applications to protect gardens and crops. Although a variety of methods and chemicals have been used to control these pests, the most commonly used preparations are bran-based powders, flakes, granules, or pellets containing metaldehyde  or methiocarb  for distribution on the ground , 10, 11.Insecticides are used worldwide, and it is inevitable that accidental poisoning of animals occur. Organophosphate and carbamates are highly toxic to all animals; some organophosphates can induce violent convulsions and kill in few minutes. These chemicals cause poisoning in animals by inhibiting the enzyme, acetylcholinesterase (AChE) which normally functions to degrade acetylcholine in nerve synapses.Figures 9, 10), can occur due to amounts of obsolete product that may be present in private farms which have not been destroyed and could be motivating farmer's intentional use against domestic and wild fauna.Recently in in Portugal the carbamate Aldicarb has been detected again compared to previous years . AldicarStrychnos nux-vomica or Strychnos ignatii, is mostly used to poison rats, birds, and wild carnivores such, foxes, and wolves. Strychnine is an extremely toxic, fast-acting poison which acts on the central nervous system leading to convulsions and ultimately death via respiratory failure. The recovery of strychnine alkaloid from the stomach contents, blood, vomitus, liver, urine, and kidneys confirms the diagnosis. Although stomach contents or vomitus have the highest concentration if death was rapid, but residues are present in liver tissues over a long time, what make this organ the ideal tissue for its detection.Strychnine, an indole alkaloid obtained from the seeds of the tree Strychnine usage and commercialization has been banned since 1988; however, strychnine continues to be one of the used substances for intentional poisoning in Portugal as results in  shows, sAnticoagulants pesticides are substances that are widely used for pest control. Poisoning due to these compounds in domestic animals  and wild animals (foxes and bird of prey) must occur by occasional consumption of anticoagulants baits and contaminated prey (secondary poisoning). Liver is the tissue of choice for detection of these molecules . These sWe are aware about the analytical technique (TLC) used because it could not allow to make an exact quantification of the substances present and because LOD could not be as low as desirable. So, it could happen that in some cases the substance could be present at concentration <LOD and be reported as undetected . If this occurs our results are lower than the real scenario of pesticide poisoning in animals in the country.In summary this study gives us a picture of the occurrence and frequency of pesticide poisoning in domestic and wild animals in Portugal since 2014 to 2020, however these results were only those obtained in LFT-FMV and probably not an exact image of the portuguese situation. In any case, as LFT-FMV is one of the most referenced laboratories nationwide by several projects taking place in country, most of the samples are sent to LFT-FMV so this study gives a good idea of the national panorama.Ingestion stills the most common way that pesticides get into non-target organisms, and most often occurs through baits left in order to kill these animals. In Europe, this illegal practice represents one of the biggest conservation problems for some endangered species, often becoming the main cause of non-natural death. Species protection projects are an asset in this matter because without them it wouldn't be possible to monitor the areas of Portugal where the protected species are located and, consequently, analyses wouldn't be carried out, and we wouldn't have a real picture of the poisonings in the country.So it is very important trying to understand what motivates these poisonings because only then will be possible to work on solutions for the problem and to take preventive and other prohibitive measures that make possible to identify who buys toxic substances and where, when and how this type of substances are acquired.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.All authors had been evolved in the work and in the manuscript elaboration.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.


Impact force remains the primary cause of foot injury and general discomfort with regard to footwear. The footwear industry traditionally relies on modified elastomers  whose properties can be physically adjusted by varying the constituents in the rubber formulations. This work aims to investigate the effect of filler/plasticizer fractions on shock attenuation of natural rubber soles. The statistical response surface method (RSM) was used to optimize the loading of natural rubber, fillers (carbon black and china clay) and a plasticizer (paraffinic oil). A novel predictive equation addressing the effects of additives on the physical and mechanical properties of the shoe sole was successfully created using the RSM. Our results demonstrate how the concentrations of these components regulate final properties, such as impact force absorption and hardness, in the commercial manufacture of shoe soles. While a higher loading level of plasticizer promotes reductions in hardness and impact force, as well as energy dissipation, in these modified elastomers, these properties were improved by increasing the filler content. Impact force constitutes the leading cause of foot injury, with pain and discomfort in both feet and legs resulting from long periods of walking or, even worse, running ,2,3,4. PA plethora of investigations has aimed at developing NR with designer properties, mostly in connection with the tire industry, by physically adjusting the content of fillers and oil. Increasing the content of carbon black (CB), for example, has been suggested as a means by which to improve the tensile properties, which are frequently related to hardness, of finished rubber, in contrast to increasing the content of a plasticizing oil ,13,14. FThe objective here is to identify the optimal loading level of three commercial additives that are used together to generate a formulation exhibiting the greatest reduction in impact force. The DoE method employed in this work will be useful in future testing and design efforts, and our findings are anticipated to be of widespread interest to developers and manufacturers of footwear, especially with regard to shoes with soles that are produced from modified NR.The NR (STR5L) rubber was purchased in block form from Chana Latex Co., Ltd. , whereas CB  and china clay  were acquired from GSP Products Co., Ltd. . The CB and CC are generally considered as reinforcement and non-reinforcement fillers, respectively. Paraffinic oil , added to NR as a plasticizer to facilitate filler dispersion, was likewise obtained from GSP Products Co., Ltd. . Other crosslinking species , and sulfur) were all provided by Kij Paiboon Chemical Co., Ltd. . 2 for 10–15 min and subsequently cooled at ambient temperature for 24 h prior to testing.Compound mixing was performed in two stages. First, the NR was masticated in a kneading machine  at 90 °C, at a roller speed of 30 rpm. Subsequently, the plasticizer and fillers were added according to the formulation in i) discern the relationships that exist between experimental inputs and (ii) optimize designs that involve multiple variables by revealing the design response to independent changes in the variables. This DoE strategy greatly reduces the number of experiments necessary in a systematic (or full-factor) analysis. The independent variables examined here, as well as their ranges of interest, are included in 1, X2, and X3 constitute the variables that are identified in Experimental design is a well-known procedure for testing hypotheses aiming to describe and possibly predict an outcome that is non-linearly dependent on multivariable conditions. The RSM algorithm used here creates a mathematical model to  according to ASTM D2240 at three different points on each specimen. The rebound PR from impact was also determined using the same instrument according to ASTM D1054. Shock absorption was evaluated according to ASTM F1614 on a custom-built drop-testing machine, as schematically depicted in 2) values are all close to unity, the predictions are consistent with the experimental results. Adjusted R2 values compare the explanatory power of the regression models that contain different numbers of predictors. These are modifications of R2 to account for the number of predictors in the model and are generally less than R2. In this study, the adjusted R2 values of each model are relatively close to the value of R2, which confirms that the model equations developed according to the RSM are appropriate. Lastly, the predicted R2 constitutes a good indicator of how well the model can predict a new  response. These values are also relatively high and not far from the adjusted R2 values, indicating that the models are also able to yield accurate property estimations. The equations that are presented in Statistical analysis is widely used to study the trend and interpretation of results from testing data. As stated in the previous section, a statistical analysis of the results obtained here from the RSM-designed experiments was conducted in conjunction with the Minitab software, and statistical significance was judged at the 95% confidence level. Predicted property responses of the three variables identified in Equation (1) are listed in The dependence of hardness on filler (CB and CC) and plasticizer (PO) loading level is linear, according to the model results that are shown in The corresponding cross-correlation surface plots of hardness with two of these additives at a time are presented for comparison in Another important property metric, the rebound PR, is commonly employed in testing elastomers to discern the energy lost or dissipated during deformation, since this resilience is inversely proportional to the energy loss. On the basis of the model coefficients that are provided in Rubber is an example of a viscoelastic material, which can be mathematically represented by a non-linear model that consists of energy-storing elastic springs and energy-dissipating viscous dashpots arranged in a wide variety of combinations  and parallel (Kelvin) models representing the most simplistic). In these frameworks, the properties of a viscoelastic material are effectively described by elements corresponding to its stiffness and damping, in which stiffness relates to hardness and damping is related to resilience. Therefore, both of these considerations influence the cushioning property of footwear. In this work, we find that the hardness of modified NR with various concentrations of additives generally follows a polynomial relationship with respect to impact force for each energy level explored, as evidenced by the results that are provided in The results of this study do not, however, reveal any direct relationship between PR and impact reduction. Because resilience is the ratio of the energy returned upon recovery from deformation to the energy of deformation, a high PR value translates into a high energy return while a low PR value indicates high energy dissipation. Because the heel is typically the first part of the foot to contact the ground, it absorbs the brunt of the impact force that is transferred to the leg and body. Therefore, the shock-absorption properties of shoe materials constitute a matter of primary importance in preventing bodily injuries due to incessant impact. The drop-test method adopted in this study provides a systematic means of measuring the shock absorption properties of NR, and the NR compound yielding the lowest peak impact force identifies the best material for shock absorption and, as a consequence, the best material for use in the production of shoe soles. Rubber with a high PR is less effective in reducing the peak impact force than rubber with a low PR, in which case we must also consider energy return as another relevant property. Prior studies ,25 have The main goal of this work is to use the RSM for optimizing the cushioning property of rubber formulations associated with impact force reduction. A DoE study based on the RSM has been performed here to establish the relationships between, as well as the optimum loading levels of, two fillers and a plasticizer in NR compounds used in the manufacture of footwear to reduce the peak impact force and, hence, foot and/or body damage. Experimental data confirms that a large level of plasticizing oil successfully reduces both the peak impact force and the hardness of rubber. On the other hand, both fillers (CB and CC) increase the hardness, thereby leading to a corresponding increase in impact force. High filler levels also promote a low PR , whereas the incorporation of the plasticizing oil has the opposite effect. On the basis of the results of this study, impact force attenuation depends on both hardness and the PR, and we have demonstrated that a reduction in impact force is associated with reduced hardness. The RSM greatly facilitates optimizing the response of the rubber compounds studied to changes in composition, and model predictions are consistent with the experimental results. This study has successfully identified the effect of commercially relevant additives in rubber formulations for footwear applications. A predictive RSM model has been developed on the basis of experimental inputs to predict the role of additives (fillers and plasticizers) on mechanical properties , which can be used to discern the most appropriate rubber formulation capable of reducing impact. Our findings verify that quadratic regression models in the context of the RSM yield the optimal loading levels of CB, CC, and PO in NR and predict the properties that correspond to a low impact force. Graphical representations displaying the relationship between impact force reduction and hardness constitute a particularly valuable tool for estimating shock absorption of rubber compounds formulated in the footwear industry. Although hardness is a relatively simple rubber property that can be easily measured, it provides tremendous insight into the ability of NR to absorb impact. Future extensions of this study should focus on other properties, such as the durability of rubber compounds and their resistance to abrasion, friction, tearing, and liquid absorption.


Studies that investigate cognitive ability in African children and estimate the general cognitive abilities of African adults tend to work with existing models of intelligence. However, African philosophy and empirical studies in cross-cultural psychology have demonstrated that conceptualizations of human cognitive ability vary with location. This paper begins with the assumption that the existing Anglo-American models of cognitive abilities are valuable but limited in their capacity to account for the various conceptualizations of valued cognitive abilities in different human societies. On the basis of this assumption, I employ extant empirical evidence generated through ethnographic studies across Africa to formulate what an African model of valued human cognitive ability ought to be. The output of this formulation has been so christened a model of valued cognitive ability in order to draw attention to the fact that models of cognitive abilities have currency and values in each human society. This value allocation is expected to influence which elements of cognitive ability each human society will promote and develop. In addition, implications for theory, research and praxes are discussed. Cognitive ability is a valuable human attribute in so far as it has been shown that it has implications for life outcomes Serpell, . Opoku , p. 538 Various studies have established that cognitive ability is a significant predictor of academic success, physical health and longevity, and job performance , Bradya et al. .real, actual and empirical worlds. It may be safe to say that intelligence operates at the real world level but the manifestations of it occur at the empirical world level of observations. The dimensions of intelligence (processes of intelligence) appear to operate at the actual world level. This means that because there are cross-cultural variations in the meaning of intelligence (processes of intelligence) at the actual world level, its measurement at the empirical world level present a slice of what intelligence may be. This is particularly problematic again when one considers Sternberg's  by capitalizing on strengths and correcting or compensating for weaknesses; (3) in order to adapt to, shape, and select environments; and, (4) through a combination of analytical, creative, and practical abilities (p. 189).If the conventional views about intelligence emphasize memory and analytical abilities, then it is difficult to assume, at the same time, that Model III thinking is correct, that the meaning of intelligence is the same across cultures. Perhaps, a better interpretation will be to consider it in terms of Bhaskar , 2008 rernberg's  definitireal world level, but its representations (dimensions or processes of intelligence) at the actual world level tends to reflect the meaning of intelligence peculiar to the particular socio-cultural context. It is against this background that the purpose of this paper is essential. Thus, the current thoughts, regardless of the empirical evidence gathered, about the constitution of intelligence is still narrow and far from being representative of how others think of intelligence.This definition may reflect intelligence at the Homo sapiens if we attempt to diversify the samples studied in psychology. This is consistent with Oppong's  but it is limited in its capacity to account for the varied representations of intelligence that other subgroups of Homo sapiens have. Thus, if the most comprehensive model of intelligence  and wisdom (nyansa). According to Gyekye .Drawing on African proverbs and customary practices in general and more specifically the Akan cosmology, Gyekye  presenteo Gyekye , p. 137,nunya”, Gyekye  wisdom. Cleverness in achieving certain ends may not amount to wisdom (p. 143).This is a clear departure from the Western models of intelligence whose validity have been supported through their relations to individual well-being Lo,  rather tThis synthesis is inspired by a similar work done by Carroll  on the cThe following inclusion criteria were used: (1) reports of empirical studies focusing on intelligence among Africans. (2) With respect to the study types, only field studies that were ethnographic or qualitative in nature or mixed methods studies were included. The study should not impose any a priori theory or conceptions of intelligence on the study participants. Studies using experimental and cross-sectional designs or tested an existing theory of intelligences were excluded. (3) Only studies that sought to identify the indigenous conceptions or implicit theory of intelligence were included. (4) Only studies published after 1990 were included. I carried out the literature search until February 22, 2020. The search terms used include the following: Africa, Sub-Saharan Africa, intelligence, cognitive abilities, and abbreviations such as IQ. The literature search was managed using the reference manager, Zotero.Four thousand seven hundred four  articles were identified from the different databases for assessment . After rThe following articles were excluded: Fortes's  study amrieko was also considered the umbrella term for the other components.The three (3) studies that were included in the analysis were published in 1993, 2001, and 2004. One was conducted in East Africa, Kenya , wisdom, and socio-emotional competence.Homo sapiens  in realizing the problem and winjo (comprehension) in working out a solution; he displays luoro (consideration and respect) for his family members, and he demonstrates rieko (skill) in generating the desired outcome (p. 370).The story is exemplary in that the boy shows The above clearly shows how Carroll's  Stratum Second, the Stratum II elements reflect common elements of the cognitive competence found in the ethnographic studies on Africans' conceptions of intelligence see . As a reThe Carroll's  Stratum the ability to think out ways of making success in one's life—to analyse and solve practical problems of life—and the ability to pay reflective attention to the fundamental principles underlying human life and experience (p. 137).Thus, wisdom is the ability to combine both cognitive competence and socio-emotional competence to successfully solve any given problem. In other words, wisdom involves demonstrating thoughtfulness, and the intellectual initiative in the realization of a problem, and having the understanding required to work out a solution while displaying consideration for one's community as well as demonstrating the skills required to generate the desired outcomes. On the other hand, socio-emotional competence relates to the display of respect and care for others, obedience towards those deserving of it, diligence, consideration, and readiness to share with one's community.Though the model is preliminary, it suggests that general intelligence or general cognitive ability, when viewed within the African context, comprises cognitive competence, wisdom, and socio-emotional competence. These three elements of cognitive abilities are interconnected. The appropriation of Carroll's  Stratum If we consider the model , wisdom and socio-emotional competence, practicing psychologists involved in assessment should interview parents and significant others about their judgement of the testees on these attributes. Another critical question to ask is: how does a clinician identify learning disabilities in children if there cannot exist common tests of cognitive abilities? The question to ask rather is: must we define learning disabilities with reference to school work? The current thinking about learning disabilities is school-based and it is said that it occurs in individuals who otherwise exhibit at least average abilities essential for thinking and or reasoning (Us National Joint Committee on Learning Disabilities, It is perhaps important to conclude that the available ethnographic data changes how we ought to conceptualize intelligence or cognitive abilities in general and indicators used to measure them. That wisdom and socio-emotional competence do not form part of Carroll's  three-stPublicly available datasets were analyzed in this study. This data can be found here: Pubmed, Web of Science, PsycINFO, Google Scholar.The author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Mesenteric cysts are rare, generally benign intra-abdominal lesions with a wide range of presentation in terms of size, clinical presentation, etiology, radiological features, and pathological characteristics.We reported a case of giant mesenteric cyst in a 16-month-old girl successfully managed in a low-resource setting.This case is particularly important not only due to the rarity of the presented case, but also for the highlighted aspects from a public health point of view.We faced of the problem of a late stage disease and the lack of preoperative diagnosis due to cultural and economic reasons and the weaknesses of healthcare systems, as in the majority of low- and middle-income countries.Despite all these limitation, this case illustrates that complex, rare diseases can also be managed successfully in a low-resource setting. It is mandatory to strengthen and improve the health system both in terms of equipment both in terms of public health policies in order to offer a better and more effective quality of care to patients also in low-income countries. In geneWe reported a case of giant mesenteric cyst successfully managed in a low-resource setting. The work has been reported in line with the SCARE criteria .2Written informed consent was obtained from the parents of the girl for publication of this case report and any accompanying images.3A 16-month-old girl presented with diffuse abdominal distention, nausea, vomiting and severe pain after one month history of abdominal discomfort. The physical examination, made difficult by painful palpation, showed severe abdominal distention with a palpable, soft and fluid-filled mass A. Plain 4This is a very rare case considering that lymphangiomas incidence ranges from 1 in 105,000 to 1 in 250,000 among admitted surgical patients and are predominant in male ,7. The cUsually, the preoperative diagnosis is done by ultrasound in order to distinguish between plain and cystic masses and computerized tomography (CT) or magnetic resonance imaging (MRI) to determine extension and cystic content . Needle Unfortunately, in our case we had no adequate diagnostic imaging and we preferred to directly choose the surgery option considering the late stage situation with noticeable symptoms. The surgical treatment is mandatory for large mesenteric cysts in order to exclude malignant transformation and to prevent complications, such as rupture, hemorrhage, torsion or infections. In case of simple lymphatic and mesothelial cysts, it is easy and curative to enucleate the mass, but lymphangiomas and benign cystic mesotheliomas could be adherent to vital intra-abdominal structures and their complete excision could be very difficult or impossible. Although limphangiomas and benign cystic mesothelioma are considered benign tumors and, thus, should be avoided the resection of vital organs, in some cases is reported a resection of different organs due to adhesion ,7. In ouRecurrence and metastasis are not frequent and occur especially if the resection is incomplete .We were able to confirm the hypnotized diagnosis first of all by macroscopic characteristics that are useful for the distinction from simple lymphatic cysts , to lymphangiomas . Moreover, we confirmed the diagnosis by the histological examination characterizing endothelial and mesothelial cells.This case is particularly important not only due to the rarity of the presented case, but also for the highlighted aspects from a public health point of view. In fact, as the result of the weaknesses of healthcare systems and also due to cultural and economic reasons, as in the majority of low- and middle-income countries we faced of the problem of a preoperative diagnosis lacking ultrasound, CT and MRI scanners. Moreover, although it is not the case, surgical and anesthesiology equipment is often poor being impossible to perform a complete preoperative or laparoscopic diagnostic procedure. In this case, an explorative laparotomy was necessary without a clear preoperative diagnosis, but this would probably not have changed outcome. In a high-resource country, a laparoscopic approach would have been used to perform the whole operation that we made by open surgery. Despite all these limitation, this case illustrates that complex, rare diseases can also be managed successfully in a low-resource setting. Unfortunately, we lost the patient in follow up due to difficult access to the hospital, living far from the hospital and due to the lack of an effective network and system for monitoring and ensuring follow-up visits.Therefore, it is mandatory to strengthen and improve the health system both in terms of equipment both in terms of public health policies in order to offer a better and more effective quality of care to patients also in low-income countries.All authors declare no conflict of interestNo funds was received for this workWritten informed consent was obtained from the parents of the child for publication of this case report and any accompanying images.Moreover the ethical approval has been exempted by our institution considering that the case was written using retrospective and anonymous data.Written informed consent was obtained from the parents of the child for publication of this case report and any accompanying images.MA: data collection and interpretation and final revision.DP: writing paper and data interpretation.MS: writing paper.ACC: data analysis and final revision.This is a case report and it is not registered.Mario Antunes.Not commissioned, externally peer-reviewed.


Our earlier article showed that increased employability of segregated Roma may improve their well-being and health. To achieve that, appropriate employment based on a public–private partnership could be the key. For optimal design of such a partnership, we need insight into its potential mechanisms. Evidence on this is lacking, however. This paper builds on the previously published article by focusing on mechanisms for achieving better health. Therefore, our aim was to identify the potential mechanisms by which a public–private Roma employment project could increase employability. We investigated a Roma employment project called Equality of Opportunity established by a private company, U.S. Steel Kosice in eastern Slovakia. We conducted a multi-perspective qualitative study to obtain key stakeholders’ perspectives on the potential mechanisms of a public–private Roma employment project in terms of increased employability. We found three types of mechanisms. The first type regarded formal job mechanisms, such as an appropriate employment and salary offer and a bottom-up approach in capacity building. The second type involved sustainability mechanisms, such as the personal profile of project and work-shift coordinators, the continuous offer of training and cooperation with relevant stakeholders . The third type was cultural mechanisms, such as personal contact with project participants, attention to less-voiced groups like children, the motivation of project participants, a counter-value reciprocity approach and respect for the specifics of Roma history. Our findings imply that policymakers could consider public–private partnerships for increasing the employability of segregated Roma, as they have the potential to address a wider range of social needs simultaneously. Roma are one of the largest ethnically delineated populations in Europe . SubstanThe low levels of employment of segregated Roma are related to both their suitability for the labor market and the structure of the labor market. Unemployment is increasingly understood to be caused by a lack of employability , which rPublic–private partnerships may be a way to improve the poor employment of segregated Roma. Public and private partners have been shown to separately lack sufficient capital to create a sustainable solution for reducing the unemployment of segregated Roma . HoweverTo achieve this, we need insight into the mechanisms that lead to the employability of segregated Roma in order to design adequate public–private partnerships. Our previous article from the same study showed that a Roma employment project based on a public–private partnership may increase the employability of segregated Roma as well as improve their well-being and health . In ordeThis paper is the result of a large study on health inequalities under the project 7FP SOPHIE , on which one paper has been published before . GeneralConceptually, we based our manuscript on a program theory for the project that we assessed , addressing the causal processes that link the implemented treatment to outcomes . For thiWe conducted a multi-perspective qualitative study investigating a Roma employment project called Equality of Opportunity, established in 2002 by U.S. Steel Kosice (USS Kosice). This project represents, in terms of size, duration, complexity and sustainability, an interesting example of an attempt to address the Roma social inequality issue. We used the Context, Mechanism, Outcome (CMO) framework to structure data collection. The CMO configuration makes up part of the realist evaluation approach  and aimsThe study consisted of several phases. The research protocol included a detailing of the data collection, i.e., the way to set up the project stages, procedures and timetable; the field procedures and data collection structure; and the questions, sample, report guide and sources of evidence. Details on the study stages and the structure of the interviews are provided in the Appendix. The second step was to collect data on the setting of the project (project background). The third step was to collect data using direct observation, in-depth semi-structured interviews, focus groups and informal face-to-face unstructured interviews. The last step was to analyze data in two rounds: the first round ran along with the data collection, and the second round was performed later. More detailed information on these stages can be found in We included the main actors in the project, i.e., the Roma community, professionals , public authorities and others , with proper methods of data collection for each group. The final sample consisted of 20 respondents  for the formal and informal interviews together, 28 respondents  for the focus groups and 98 respondents (gender not monitored) for direct observation .We collected data using in-depth semi-structured interviews, informal unstructured interviews, focus groups and direct observation. We carried out the in-depth semi-structured interviews using a predefined set of topics and open-ended key questions, with the aim of systematically covering all topics of interest . We performed these interviews face-to-face, collecting the data by audio recording (with informed consent from participants) and by written field-notes. The layout and structure of the interviews are presented in We carried out the informal unstructured interviews to gain the views of people not directly involved in but possibly affected by the project. These interviews were done by one researcher accompanied by a Roma community worker who, if necessary, also translated from and to the Romani language. The researcher collected the data using written field-notes.We conducted focus groups using a predefined set of topics and open-ended key questions to gain the views of people not directly involved in but possibly affected by the project. We conducted three focus groups, each by three researchers, with the contents of interviews collected by written notes. The focus group with Roma children was facilitated by a Roma assistant who, if necessary, also translated from and to the Romani language. The layout and structure of the focus group scenario are presented in Finally, we used non-participatory direct observation without interaction during the recruitment process to examine participants, their settings and their practices when applying for a job in the project. During this recruitment, we also observed the attitudes and habits of USS Kosice representatives. Furthermore, we directly visited the residences of participants in an effort to capture life in the settlements. For this, we used direct observation of participants with interaction. During the visits to the settlements, we were without USS Kosice representatives, accompanied only by local community workers who, if necessary, also translated from and to the Romani language.We collected data on the setting of the project, the characteristics of the participants and on potential mechanisms by which the project could increase employability. Regarding the project setting, we collected data on the project launch, including its circumstances and key personnel, the project’s main goal, the type of contracts, financial remuneration and the characteristics of the project participants. Regarding these characteristics, we assessed gender, locality, age and education. Age structure, education level and average number of workers were calculated based on the overall number of participants since the start of the project in 2002. We further collected data using the CMO framework on potential mechanisms by which the project could increase employability. We understood these mechanisms to be measures  that triggered specific outcomes within a particular context. This regards only mechanisms/measures from the CMO framework. Examples of questions regarding the mechanisms of the CMO framework are shown in First, we described the project setting (the project background). Second, we assessed the mechanisms potentially leading to increased employability of segregated Roma using the CMO framework. For this second step, we performed a content analysis of the data based on recurrent abstraction, i.e., repetition of reading and summarizing in steps of data coded as relating to the same topics or variables . The conThe study was conducted in the setting of the Equality of Opportunity project. This project was financed by USS Kosice and carried out in cooperation with the municipalities comprising the adjacent Roma settlements. USS Kosice, an integrated steel producer, is a subsidiary of the United States Steel Corporation headquartered in Pittsburgh, USA, and the largest private employer in eastern Slovakia. The project was initiated in 2002 by the first president of USS Kosice, who offered job vacancies in response to a request for support to reduce a number of problems  in the nearby Roma settlement. USS Kosice offered Roma jobs with a significantly higher salary than the minimum wage and with training, while municipalities covered the selection of the candidates. Successful candidates formally became employees of the municipality but were assigned to USS Kosice for temporary work. USS Kosice has created around 170 jobs for segregated Roma since 2002 (having 0 jobs of this kind before this initiative). More extensive information can be found in Bosakova (2018) [Project participants were all males and came mainly from three settlements that were within a 15 km radius of the USS Kosice plant. Ages of participants ranged from 18 to 60 years, and nearly three quarters were aged 21–40 years. Most project participants had completed only primary school .Informants perceived three types of mechanisms that lead to an increase in the employability of segregated Roma. The first type concerned formal job mechanisms , with suThe second type of mechanism involved sustainability mechanisms , with suThe third type of mechanism involved cultural mechanisms , with suWe explored the potential mechanisms of a public–private Roma employment project in terms of increased employability. As mentioned in the Results, we found three types of mechanisms. The first type was formal job mechanisms, such as an appropriate employment and salary offer and a bottom-up approach in capacity building. The second type involved sustainability mechanisms, such as the personal profile of the project and work shift coordinators and the continuous offer of training and cooperation with the relevant stakeholders  Regular monthly meetings and agreement on temporary work assignments  were also mentioned but only by professionals and public authorities. The third type consisted of cultural mechanisms, such as personal contact with project participants, attention to less-voiced groups such as children, motivation of project participants, a counter-value reciprocity approach and respect for the specifics of Roma history. Stakeholders considered these three types of mechanisms to lead to an increase in the employability of segregated Roma. We will further discuss these results, i.e., the three main groups of mechanisms, in more detail.Informants mentioned appropriate work and salary offer and a bottom-up approach to capacity building as the formal job mechanisms that led to the better employability of segregated Roma. In terms of increased employability, this group of mechanisms may help them gain work experience, acquire skills and improve knowledge and motivation to maintain employment. An appropriate work offer is aligned with the current capabilities of this workforce but does not have to be only menial , and togInformants further mentioned a number of mechanisms, such as the personal profile of the project and work shift coordinators, the continuous offer of training and cooperation with relevant stakeholders , which may contribute not only to the sustainability of the project but also to the sustainability of the participants’ involvement in the project. In terms of increased employability, this group of mechanisms can help develop personal attributes and understanding, improve knowledge and enhance skills also beyond work. The coordinator’s profile in terms of being a person with adequate capacity and skills is one of the critical success factors . MusinkaRegular monthly meetings and agreement on temporary work assignments  were also mentioned as promoting sustainability but only by professionals and public authorities. Regular monthly meetings provide coordinators room for evaluation, enable better monitoring of what works  and provInformants further mentioned a number of mechanisms related to culture that could enhance the success of public–private partnerships in relation to the employability of segregated Roma, such as personal contact with project participants, attention for less-voiced groups like children, motivation of project participants and respect for the specifics of Roma history. In terms of increased employability, this group of mechanisms may help to develop personal attributes, improve knowledge and understanding and increase the motivation to gain and maintain employment.Personal contact, as a mechanism that may increase the employability of segregated Roma, is crucial, when we consider the stigmatization of Roma outside the settlement and the often tense relationships between Roma and non-Roma . This suInformants also mentioned attention for less-voiced groups like children as a mechanism related to increasing the employability of segregated Roma. This mechanism is based on the life course way of thinking and includes close cooperation with local primary schools  but also the involvement of children in various projects, attempting to motivate them to complete primary education and continue their studies at least at partner vocational schools . This isMotivation, a counter-value reciprocity approach and respect for the specifics of Roma history were seen as other mechanisms that may help to increase the employability of segregated Roma. Non-Roma often act towards the Roma in a discriminatory and racist manner, and even honest attempts to provide support to Roma, typically drawn on various misinformed concepts, are practically inapplicable . MotivatThis study addressed the mechanisms by which a public–private Roma employment project could increase employability, departing from a constructed program theory  of the pOur study has several strengths, the most important one being its wide range of informants and sources, which enabled various perspectives to be identified. This approach increased the robustness and transferability of the findings. However, some limitations need to be mentioned. We used a qualitative design for analyzing this particular best practice, which does not allow generalization to other communities. We think, however, that our findings represent a typical example of the issues related to highly deprived groups like Roma. Furthermore, social desirability may have affected the responses. However, we used a multi-informant strategy to get a full picture.The results of our study have implications for improving social policies. They imply that policymakers could consider public–private partnerships for increasing the employability of segregated Roma but with an emphasis on ensuring transparency to prevent any abuse. Such partnerships could provide gains in equal work and salary opportunities for all, regardless of ethnicity. Further, gradual capacity building via a bottom-up approach and training with respect to the specifics of Roma history may also be considered when designing similar projects. In addition, policy-makers should address a wider range of social needs simultaneously by reinforcing mutual cooperation with the relevant stakeholders, such as municipalities, community centers and other NGOs. Thus, they need to coordinate simultaneously across different fields covering not only employment but also education, housing, health and family, as the employability of individuals is often interrelated with other dimensions as well as with the employability of their family and wider communities, which need to be addressed as well.A next step in future research may be to develop a longitudinal quantitative study to assess the actual impact of such a program over time, as well as the mediators of these effects and the degree to which various contextual factors moderate these associations. Future research could also assess more in-depth mechanisms on the employability of Roma in other regions or settings, which may yield large gains in the employment of this deprived group.We conclude that potential mechanisms exist within a public–private Roma employment project that may help to increase employability of segregated Roma. The identified mechanisms are related to formal job mechanisms, such as appropriate and equal employment and salary opportunities, and gradual capacity building, which together could help segregated Roma gain work experience and skills and improve their knowledge and motivation to maintain employment. There are also sustainability mechanisms, such as the personal profile of the project and work shift coordinators, an offer of continuous training and cooperation with the relevant stakeholders , which may help to develop personal attributes and understanding, improve knowledge and also develop skills other than working skills. Lastly, there are also cultural mechanisms, which may help segregated Roma develop their personal attributes and improve their knowledge and understanding and increase the motivation to gain and maintain employment. These relate to personal contact with participants, attention for less-voiced groups like children, motivation of participants, a counter-value reciprocity approach and respect for the specifics of Roma history. The identified mechanisms could help increase the employability of segregated Roma and thus their actual employment, which may help improve their living situation in a disadvantaged setting.