The United States (U.S.) government has been attempting to combat the growing opioid epidemic ravaging the nation. The opioid epidemic has had a significant impact on public health and forensic science laboratories. Moreover, this epidemic has moderate to fatal health consequences for expectant mothers with substance use disorder and their child who may develop Neonatal Abstinence Syndrome (NAS), otherwise known as Neonatal Opioid Withdrawal Syndrome (NOWS). The objective of this thesis is to emphasize that further research is needed for the identification and quantification of opioids in human breast milk. This topic has public health implications such as discussing the information gaps as it relates to a highly vulnerable group, women, and infants, affected by the opioid epidemic. Furthermore, there are implications in forensic science connected to postmortem toxicology and pathology when determining the cause of death and contributing factors in pediatric cases. This emphasis on the need for greater research will be accomplished by highlighting the opioid epidemic, its impact and further understanding of the addictive drug class known as opioids. The history of the crisis, effects on society as well as pharmaceutical knowledge of opioids will assist in development of plans to suppress growth and provide care for the afflicted. Furthermore, this thesis will attempt to demonstrate the need for further research involving opioids will be of significant value for public health and forensic science. As the forensic laboratories and various medical facilities are at the forefront of the opioid epidemic, there is a need for more robust, validated, inexpensive, and fast drug detection methodologies. Increasing rates of new designer drugs, addiction, and opioid-related deaths has caused a backlog in the forensic laboratories due to the great number of cases. While, the higher instances of maternal substance use disorder (SUD)/ opioid use disorder (OUD) with parallel increases in cases of NAS incidences are a few of the issues that need to be managed by public health leaders. Additionally, this thesis will examine current methodologies for drug quantification of opioids in human breast milk. The valid methodologies developed as well as the findings by the few available studies allowed for the current recommendations related to the acceptability of mothers in MAT programs, using methadone and buprenorphine during pregnancy and postpartum, being able to breastfeed their infant. By examining these studies and the findings, standardization criteria for the development of study designs for new methodologies relating to drug determination in human breast milk could be developed. The establishment of standardization criteria and acknowledging information gaps in current knowledge will be significant as these findings could influence policies, guidelines and procedures relating to maternal SUD/OUD, NAS/NOWS, and pediatric death determination as well as postmortem toxicology.

Breastfeeding is associated with long-term wellbeing including low risks of infectious diseases and non-communicable diseases such as asthma, cancer, autoimmune diseases and obesity during childhood. In recent years, important advances have been made in understanding the human breast milk (HBM) composition. Breast milk components such as, non-immune and immune cells and bioactive molecules, namely, cytokines/chemokines, lipids, hormones, and enzymes reportedly play many roles in breastfed newborns and in mothers, by diseases protection and shaping the immune system of the newborn. Bioactive components in HBM are also involved in tolerance and appropriate inflammatory response of breastfed infants if necessary. This review summarizes the current literature on the relationship between mother and her infant through breast milk with regard to disease protection. We will shed some light on the mechanisms underlying the roles of breast milk components in the maintenance of health of both child and mother.

Studies of the interaction of the relevant metal, metalloid or nanoparticulate species with biological systems are underpinned by the provision of reliable information about chemical composition of the relevant materials. Often, no methods of chemical analysis are available. The work described in this dissertation centers on developing methods to help with studies for a variety of analytes and samples. A method was developed for the determination of 11 trace elements (As, Cd, Co, Cr, Cu, Fe, Mg, Mn, Pb, Ti, and Zn) in human breast milk and infant formulas by inductively coupled plasma optical emission spectrometry (ICP-OES) following microwave-assisted digestion. A method was established for the determination of trace elements, with an emphasis on titanium as titanium dioxide, in snack foods and consumer products. The interactions of some dissolved metals, including rare earth elements, and metallo-nanoparticles (silver, gold, titanium dioxide, aluminum oxide, and iron) with aquatic plants were studied. After exposure in a variety of mesocosms, the partitioning of the elemental species between various compartments was quantified by ICP-MS and ICP-OES following microwave-assisted digestion. An ICP mass spectrometry (MS) method has also been developed to quantify the uptake of gold and silver nanoparticles by C. elegans. Uptake of gold nanoparticles was size dependent, suggesting increased ingestion efficiency with increased particle diameter. The feasibility of discriminating between suspended TiO2 nanoparticles and dissolved titanium by the analysis of the rapid transient signal events obtained from the ICP-MS instrument operated in a rapid response mode was also developed. Data handling parameters were established that allowed a distinction in the signals for nanoparticulate and standard solutions. Spikes in the signal were defined by distinct parameters using the mean and standard deviation, where a spike in the signal was defined as a signal > + ks (k =3). This approach however did lead to a statistical difference in the spike signal events for solutions and nanosuspensions.

As aflatoxinas são substâncias naturais que apresentam efeitos tóxicos aos humanos e são reconhecidamente carcinogênicas. Estas substâncias podem estar presentes na dieta humana ou, em casos específicos, no ar respirado. Desta maneira, a exposição humana às aflatoxinas é objeto de muita preocupação. Uma das maneiras mais eficazes de avaliar a exposição humana as aflatoxinas é através da mensuração da presença de biomarcadores da exposição a estas substâncias em fluidos biológicos. Dentre as possibilidades de biomarcadores de exposição às aflatoxinas tem-se que aflatoxina M1 (AFM1), presente na urina e leite humano, é considerada um biomarcador válido. Assim sendo, o objetivo deste trabalho de pesquisa foi avaliar a presença de AFM1 em amostras de urina provenientes de indivíduos residentes na região urbana e rural da cidade de Piracicaba-SP, assim como, de leite de gestantes de Piracicaba e cidades da região. Nos indivíduos doadores de amostras de urina foi levantado também o padrão de ingestão de alimentos com alto risco de conter aflatoxinas, através da aplicação de inquéritos de freqüência alimentar e recordatórios 24 horas. A análise de AFM1 em urina e leite foi realizada por cromatografia liquida de alta eficiência (CLAE) com detecção por fluorescência. A extração e purificação do extrato foram realizadas com auxílio de colunas de imunoafinidade. No total 69 amostras de urina e 18 de leite foram analisadas. Entre as amostras de urina detectou-se a presença de AFM1 em 54 (78%) das amostras, com concentrações variando de 1,8 até 39,9 pg/mL. Não foi observada diferença estatística entre as concentrações médias detectadas entre urinas de indivíduos da zona urbana e rural, bem como no nível de consumo de produtos de risco. Apesar das concentrações de AFM1 detectadas serem inferiores as concentrações médias reportadas em outros países a freqüência de amostras positivas foi bastante elevada mostrando que as populações estudadas estão sendo expostas às aflatoxinas. Assim, melhores avaliações dos níveis de exposição necessitam ser realizados considerando que a amostragem utilizada foi pontual, pode existir variação de contaminação sazonal com aflatoxinas na dieta e a contaminação é heterogênea dentro no alimento. Não foi observada uma correlação entre o nível do consumo de produtos de risco e as concentrações detectadas em amostras de urina. Apenas uma amostra de leite apresentou contaminação detectada; entretanto, o nível de contaminação estava entre o limite de detecção (LD) e o limite de quantificação (LQ). / Aflatoxins are natural substances that present toxic and carcinogenic effects to humans. These substances may be present in human diet or, in specific cases, in the breathing air. Thus, the human exposition to aflatoxins is object of concern. One of the most effective ways to evaluate human exposition to aflatoxins is to measure the presence of biomarkers in biological fluids. Among the possibilities of aflatoxin presence biomarkers, the aflatoxin M1 (AFM1), present in human urine and milk, is considered a valid biomarker. The objective of this work was to evaluate the presence of AFM1 in urine samples from individuals who live in urban and rural areas in the county of Piracicaba, state of São Paulo, Brazil, and in milk of pregnant women from Piracicaba and neighbor cities. Urine-donor individuals were researched in relation to the ingestion of food with high risk of containing aflatoxins through the application of a food frequency questionnaire and 24-hour recall. The analysis of AFM1 in urine and milk was performed through high-performance liquid chromatography (HPLC) with fluorescence detection. The extract purification and extraction were performed with the aid of immunoaffinity columns. Overall, 69 urine and 18 human breast milk samples were analyzed. Among urine samples, the presence of AFM1 was detected in 54 (78%), with concentrations ranging from 1.8 to 39.9 pg/mL. No statistical difference was observed between average concentrations detected in the urine of individuals from urban and rural areas, as well as the consumption of aflatoxin risky food. Although the AFM1 concentrations detected are lower than those reported for other countries, the frequency of positive samples was quite high, showing that the populations studied are exposed to aflatoxins. Thus, further evaluations on the exposition levels should be performed, and considering that the sampling used in this work was punctual, there may be seasonal contamination variations in diet and the contamination level is heterogeneous within a food. No correlation between the consumption of risky food and concentrations detected in urine samples was observed. Only one milk sample presented detected contamination; however, the contamination level was between the limit of detection (LOD) and the limit of quantification (LOQ).

As aflatoxinas são substâncias naturais que apresentam efeitos tóxicos aos humanos e são reconhecidamente carcinogênicas. Estas substâncias podem estar presentes na dieta humana ou, em casos específicos, no ar respirado. Desta maneira, a exposição humana às aflatoxinas é objeto de muita preocupação. Uma das maneiras mais eficazes de avaliar a exposição humana as aflatoxinas é através da mensuração da presença de biomarcadores da exposição a estas substâncias em fluidos biológicos. Dentre as possibilidades de biomarcadores de exposição às aflatoxinas tem-se que aflatoxina M1 (AFM1), presente na urina e leite humano, é considerada um biomarcador válido. Assim sendo, o objetivo deste trabalho de pesquisa foi avaliar a presença de AFM1 em amostras de urina provenientes de indivíduos residentes na região urbana e rural da cidade de Piracicaba-SP, assim como, de leite de gestantes de Piracicaba e cidades da região. Nos indivíduos doadores de amostras de urina foi levantado também o padrão de ingestão de alimentos com alto risco de conter aflatoxinas, através da aplicação de inquéritos de freqüência alimentar e recordatórios 24 horas. A análise de AFM1 em urina e leite foi realizada por cromatografia liquida de alta eficiência (CLAE) com detecção por fluorescência. A extração e purificação do extrato foram realizadas com auxílio de colunas de imunoafinidade. No total 69 amostras de urina e 18 de leite foram analisadas. Entre as amostras de urina detectou-se a presença de AFM1 em 54 (78%) das amostras, com concentrações variando de 1,8 até 39,9 pg/mL. Não foi observada diferença estatística entre as concentrações médias detectadas entre urinas de indivíduos da zona urbana e rural, bem como no nível de consumo de produtos de risco. Apesar das concentrações de AFM1 detectadas serem inferiores as concentrações médias reportadas em outros países a freqüência de amostras positivas foi bastante elevada mostrando que as populações estudadas estão sendo expostas às aflatoxinas. Assim, melhores avaliações dos níveis de exposição necessitam ser realizados considerando que a amostragem utilizada foi pontual, pode existir variação de contaminação sazonal com aflatoxinas na dieta e a contaminação é heterogênea dentro no alimento. Não foi observada uma correlação entre o nível do consumo de produtos de risco e as concentrações detectadas em amostras de urina. Apenas uma amostra de leite apresentou contaminação detectada; entretanto, o nível de contaminação estava entre o limite de detecção (LD) e o limite de quantificação (LQ). / Aflatoxins are natural substances that present toxic and carcinogenic effects to humans. These substances may be present in human diet or, in specific cases, in the breathing air. Thus, the human exposition to aflatoxins is object of concern. One of the most effective ways to evaluate human exposition to aflatoxins is to measure the presence of biomarkers in biological fluids. Among the possibilities of aflatoxin presence biomarkers, the aflatoxin M1 (AFM1), present in human urine and milk, is considered a valid biomarker. The objective of this work was to evaluate the presence of AFM1 in urine samples from individuals who live in urban and rural areas in the county of Piracicaba, state of São Paulo, Brazil, and in milk of pregnant women from Piracicaba and neighbor cities. Urine-donor individuals were researched in relation to the ingestion of food with high risk of containing aflatoxins through the application of a food frequency questionnaire and 24-hour recall. The analysis of AFM1 in urine and milk was performed through high-performance liquid chromatography (HPLC) with fluorescence detection. The extract purification and extraction were performed with the aid of immunoaffinity columns. Overall, 69 urine and 18 human breast milk samples were analyzed. Among urine samples, the presence of AFM1 was detected in 54 (78%), with concentrations ranging from 1.8 to 39.9 pg/mL. No statistical difference was observed between average concentrations detected in the urine of individuals from urban and rural areas, as well as the consumption of aflatoxin risky food. Although the AFM1 concentrations detected are lower than those reported for other countries, the frequency of positive samples was quite high, showing that the populations studied are exposed to aflatoxins. Thus, further evaluations on the exposition levels should be performed, and considering that the sampling used in this work was punctual, there may be seasonal contamination variations in diet and the contamination level is heterogeneous within a food. No correlation between the consumption of risky food and concentrations detected in urine samples was observed. Only one milk sample presented detected contamination; however, the contamination level was between the limit of detection (LOD) and the limit of quantification (LOQ).

Hintergrund Ein rascher enteraler Nahrungsaufbau bei Frühgeborenen verkürzt die Zeit der parenteralen Ernährung. Somit lassen sich bestimmte Risikofaktoren beeinflussen, die evt. die Morbiditätshäufigkeit der Kinder senken könnten. Mehrere Kohortenstudien zeigten, dass ein standardisierter Nahrungsaufbau mit einer geringeren Komplikationsrate und einem schnelleren Nahrungsaufbau assoziiert ist. Ziel der Studie ist zu überprüfen, ob ein standardisiertes Ernährungsprogramm einen rascheren und komplikationsärmeren enteralen Nahrungsaufbau bei Frühgeborenen ermöglicht. Patienten und Methode In die vorliegende randomisierte, kontrollierte Studie wurden 99 Frühgeborene mit einem Geburtsgewicht von ≤1750 g aufgenommen. In der Gruppe mit standardisierter Ernährung (ST) wurde der enterale Nahrungsaufbau mit Muttermilch oder gespendeter Frauenmilch nach einem speziell ausgearbeiteten Protokoll durchgeführt. In der Gruppe mit der individuellen Ernährungsform (IN) wurde je nach Bedarf und Zustand des Kindes auch semi-elementare Nahrung (Pregomin®) gefüttert. Über die Steigerungsdynamik und Nahrungspausen wurde hier individuell entschieden. Primäres Zielkriterium war die Dauer bis zum Erreichen der vollenteralen Ernährung. Ergebnisse In der ST-Gruppe war die vollständig enterale Ernährung nach 14,93 ± 9,95 (Median 12) Tagen, in der IN-Gruppe nach 16,23 ± 10,86 (Median 14) Tagen möglich. Es konnte kein signifikanter Unterschied gefunden werden. Nur bei hypotrophen Frühgeborenen erwiesen sich die Unterschiede bei der ST-Gruppe gegenüber der IN-Gruppe als statistisch signifikant: 10,20 ± 4,78 (Median 8,5) vs. 16,73 ± 8,57 (Median 15) Tage (p = 0,045). Die Gewichtsentwicklung verlief in beiden Studiengruppen nicht different. Die Kinder in der ST-Gruppe konnten bei einem Gewicht von 116% des Geburtsgewichtes vollständig enteral ernährt werden, in der IN-Gruppe bei einem Gewicht von 122% des Geburtsgewichtes. Die Inzidenz der nekrotisierenden Enterokolitis (4%) und anderer Komplikationen blieb in beiden Studiengruppen niedrig. Die Diagnose „Ernährungsschwierigkeiten“ wurde mit klaren Symptomen definiert und in der IN-Gruppe doppelt so oft gestellt, wie in der ST-Gruppe (14 vs. 7) Schlussfolgerung Das Standardisieren führte unter den Studienbedingungen nicht zu einer Beschleunigung des Nahrungsaufbaus. Anhand unserer Ergebnisse ist es möglich, dass die hypotrophen Frühgeborenen von der standardisierten Ernährung entsprechend des Ernährungsprotokolls profitieren. Diese Hypothese muss in einer neuen Studie überprüft werden. Diese Kinder konnten schneller vollständig enteral ernährt werden, als Frühgeborene, mit individuellem enteralem Nahrungsaufbau. Ein standardisiertes Nahrungsprotokoll ist im klinischen Alltag durchsetzbar, und darauf aufbauend ein enteraler Nahrungsaufbau unter strenger klinischer Beobachtung ohne Komplikationen erfolgreich durchführbar.

Hintergrund Ein rascher enteraler Nahrungsaufbau bei Frühgeborenen verkürzt die Zeit der parenteralen Ernährung. Somit lassen sich bestimmte Risikofaktoren beeinflussen, die evt. die Morbiditätshäufigkeit der Kinder senken könnten. Mehrere Kohortenstudien zeigten, dass ein standardisierter Nahrungsaufbau mit einer geringeren Komplikationsrate und einem schnelleren Nahrungsaufbau assoziiert ist. Ziel der Studie ist zu überprüfen, ob ein standardisiertes Ernährungsprogramm einen rascheren und komplikationsärmeren enteralen Nahrungsaufbau bei Frühgeborenen ermöglicht. Patienten und Methode In die vorliegende randomisierte, kontrollierte Studie wurden 99 Frühgeborene mit einem Geburtsgewicht von ≤1750 g aufgenommen. In der Gruppe mit standardisierter Ernährung (ST) wurde der enterale Nahrungsaufbau mit Muttermilch oder gespendeter Frauenmilch nach einem speziell ausgearbeiteten Protokoll durchgeführt. In der Gruppe mit der individuellen Ernährungsform (IN) wurde je nach Bedarf und Zustand des Kindes auch semi-elementare Nahrung (Pregomin®) gefüttert. Über die Steigerungsdynamik und Nahrungspausen wurde hier individuell entschieden. Primäres Zielkriterium war die Dauer bis zum Erreichen der vollenteralen Ernährung. Ergebnisse In der ST-Gruppe war die vollständig enterale Ernährung nach 14,93 ± 9,95 (Median 12) Tagen, in der IN-Gruppe nach 16,23 ± 10,86 (Median 14) Tagen möglich. Es konnte kein signifikanter Unterschied gefunden werden. Nur bei hypotrophen Frühgeborenen erwiesen sich die Unterschiede bei der ST-Gruppe gegenüber der IN-Gruppe als statistisch signifikant: 10,20 ± 4,78 (Median 8,5) vs. 16,73 ± 8,57 (Median 15) Tage (p = 0,045). Die Gewichtsentwicklung verlief in beiden Studiengruppen nicht different. Die Kinder in der ST-Gruppe konnten bei einem Gewicht von 116% des Geburtsgewichtes vollständig enteral ernährt werden, in der IN-Gruppe bei einem Gewicht von 122% des Geburtsgewichtes. Die Inzidenz der nekrotisierenden Enterokolitis (4%) und anderer Komplikationen blieb in beiden Studiengruppen niedrig. Die Diagnose „Ernährungsschwierigkeiten“ wurde mit klaren Symptomen definiert und in der IN-Gruppe doppelt so oft gestellt, wie in der ST-Gruppe (14 vs. 7) Schlussfolgerung Das Standardisieren führte unter den Studienbedingungen nicht zu einer Beschleunigung des Nahrungsaufbaus. Anhand unserer Ergebnisse ist es möglich, dass die hypotrophen Frühgeborenen von der standardisierten Ernährung entsprechend des Ernährungsprotokolls profitieren. Diese Hypothese muss in einer neuen Studie überprüft werden. Diese Kinder konnten schneller vollständig enteral ernährt werden, als Frühgeborene, mit individuellem enteralem Nahrungsaufbau. Ein standardisiertes Nahrungsprotokoll ist im klinischen Alltag durchsetzbar, und darauf aufbauend ein enteraler Nahrungsaufbau unter strenger klinischer Beobachtung ohne Komplikationen erfolgreich durchführbar.