[Melatonin reverses oxidative damage in the submandibular gland of rats treated with Cyclophosphamide]. / Melatonina revierte el daño oxidativo en glándula submandibular de ratas tratadas con Ciclofosfamida.

Objetive: Cyclophosphamide (Cf) produces oxidative damage in rat submandibular gland (GSM). In the present work we evaluated the antioxidant protective effect of melatonin (MLT) in GSM of rats treated with Cf. Methods: 40 adult male Wistar rats were divided into 5 groups (G): G1: control; G2: Control+Ethanol: treated with 1% ethanol for 10 consecutive days. On days 11 and 12 they received a dose of saline; G3: Cf: treated with 1% ethanol for 12 days, days 11 and 12 they received an intraperitoneal (i.p.) dose of Cf 50 mg/Kg/kg of saline. ) of Cf 50 mg/kg bw; G4: Cf + MLT: MLT (5 mg/kg bw, intraperitoneal, dissolved in 1% ethanol) was administered daily, days 11 and 12 received Cf same as G3; G5: MLT: treated 12 consecutive days with MLT (same dose as G4). After 12 hours of fasting, animals were anesthetized to obtain both submandibular glands, then they were sacrificed. Uric acid (UA), lipid peroxides (LPs), aqueous peroxides (APs) and superoxide dismutase (SOD) activity were measured in submandibular gland homogenate. Statistical analysis: we used ANOVA and Bonferroni test pos hoc, considering significant p<0.05. Results: Cf treatment decreased AU concentration and SOD activity (AU, mg/mg prot., G1: 2.50±0.68; G2: 2.18±0.13; G3: 0.54±0.09* G4: 1.95±0.24#, G5: 2.64±0.47, *p<0.01 G3 vs G1, G2, G4; #p<0.01 G4 vs G3 and G5; SOD, U/mg prot, G1: 4.57±0.95, G2: 4.79±0.94, G3: 2.18±0.53*, G4: 5.13±1.10, G5: 5.09±0.39, *p< 0.01 G3 vs G1, G2, G4 and G5). MLT treatment prevented these effects. In addition, Cf increased PL and PA formation. Conclusion: MLT improved the redox status in GSM of Cf-treated rats. MLT could prevent oxidative processes in GSM produced by Cf.

Objetivo: Ciclofosfamida (Cf) produce daño oxidativo en glándula submandibular (GSM) de ratas. En el presente trabajo se evaluó el efecto protector antioxidante de melatonina (MLT) en GSM de ratas tratadas con Cf. Método: Se utilizaron 40 ratas Wistar machos adultas divididas en 5 grupos (G): G1: control; G2: Control+Etanol: tratados con etanol al 1% durante 10 días consecutivos. Los días 11 y 12 recibieron una dosis de solución salina; G3: Cf: tratados con etanol al 1% durante 12 días, días 11 y 12 recibieron una dosis intraperitoneal (i.p.) de Cf de 50 mg/Kg de pc; G4: Cf + MLT: se administró diariamente MLT (5 mg/Kg pc, intraperitoneal, disuelta en etanol al 1%), días 11 y 12 recibieron Cf igual que G3; G5: MLT: tratamiento 12 días consecutivos con MLT (igual dosis de G4). Los animales fueron anestesiados, extirpándose ambas GSM y sacrificados, previo ayuno 12 hs. Se midió la concentración de ácido úrico (AU), peróxidos lipídicos (PL) y acuosos (PA) y actividad de superóxido dismutasa (SOD) en homogenato de GSM. Análisis estadístico: ANOVA y test de bonferroni, considerando significativo p<0,05. Resultados: El tratamiento con Cf disminuyó la concentración de AU y la actividad de SOD (AU, mg/mg prot., G1: 2,50±0,68; G2: 2,18±0,13; G3: 0,54±0,09* G4: 1,95±0,24#, G5: 2,64±0,47, *p< 0,01 G3 vs G1, G2, G4; #p< 0,01 G4 vs G3 y G5; SOD, U/mg prot., G1: 4,57±0.95, G2: 4,79±0,94, G3: 2,18±0,53*, G4: 5,13±1,10, G5: 5,09±0,39, *p< 0,01 G3 vs G1, G2, G4 y G5). El tratamiento con MLT previno esos efectos. Además, Cf aumentó la formación PL y PA. Conclusión: MLT mejoró el estado redox en GSM de ratas tratadas con Cf. MLT podría prevenir los procesos oxidativos en GSM producidos por Cf.

[Determination of some oxidative stress, functional and immunological markers in the saliva of patients undergoing bone marrow transplantation (BMT).] / Determinación de algunos marcadores de estrés oxidativo, funcionales e inmunológicos en saliva de pacientes sometidos a trasplante de médula ósea (TMO).

OBJECTIVE: High doses of chemotherapy used prior to bone marrow transplantation (BMT) promote severe changes in the stomatognathic system. The objective of the present work consisted in evaluating some functional, immunological and oxidative stress markers in saliva of these patients. METHODS: A longitudinal observational study was carried out on 22 patients admitted to the Bone Marrow Transplant Unit of the Oncohematology Service of the Sanatorio Allende between March 2019 and February 2020. Basal saliva collection was carried out in the initial stage (I ) prior to isolation and middle stage (M) 14 days after conditioning therapy and transplantation. The concentration of uric acid (UA), superoxide dismutase (SOD), malondialdehyde (MDA), salivary alpha amylase, secretory immunoglobulin A (Ig As), lactoferrin, ceruloplasmin and urea were analyzed. RESULTS: In (M) the levels of SOD and MAD increased significantly compared to (I) (p <0.01). The concentration of salivary alpha amylase, Ig As, lactoferrin and uric acid was significantly lower in (M) compared to (I ) p <0.0001, p <0.01, p <0.0001, p <0.02 respectively. Ceruloplasmin and Urea did not show variations during treatment. CONSLUSION: In the present study, a decrease in the defensive capacity of saliva was observed as a consequence of a reduction in the concentration of Ig As and lactoferrin. The increase in SOD in (M) could be interpreted as a defense mechanism of saliva against oxidative stress produced by chemotherapy. The decrease in uric acid in stage (M) could allow the worsening of mucositis. The synthesis and release of amylase was affected by treatment with cytostatic drugs.

OBJETIVO: Altas dosis de quimioterapia utilizadas previo al trasplante de médula ósea (TMO) pueden promover severos cambios en el sistema estomatognático. El objetivo consistió en evaluar algunos marcadores funcionales, inmunológicos y de estrés oxidativo en saliva de pacientes sometidos a dicho tratamiento. MÉTODOS: Estudio observacional longitudinal en 22 pacientes de la Unidad de trasplante de Médula Ósea del Servicio de Oncohematología del Sanatorio Allende. Se efectuó recolección de saliva basal en etapa inicial (I) previa al aislamiento y etapa media (M) 14 días posteriores a la terapia de acondicionamiento y trasplante. Se analizó la concentración de ácido úrico (AU), superóxido dismutasa (SOD), malondialdehido (MDA), alfa amilasa salival, inmunoglobulina A secretora (Ig As), lactoferrina, ceruloplasmina y urea. RESULTADOS: En (M) los niveles de SOD y MAD aumentaron significativamente respecto de (I) (p< 0.01). La concentración de  alfa amilasa salival, Ig As, lactoferrina y ácido úrico fue significativamente menor en (M) respecto de ( I ) p < 0.0001, p < 0.01, p < 0.0001, p <0.02 respectivamente. Ceruloplasmina y Urea no mostraron variaciones. CONCLUSIÓN: se observó una disminución de la capacidad defensiva de la saliva como consecuencia de una reducción de la concentración de Ig As y lactoferrina. El incremento de SOD en (M) podría interpretarse como un mecanismo de defensa de la saliva contra el estrés oxidativo producido por la quimioterapia. La disminución de ácido úrico en la etapa (M) podría favorecer el agravamiento de mucositis.  La síntesis y liberación de amilasa fue afectada por el tratamiento con citostáticos.

Efecto de carboplatino sobre malondialdehído como marcador de lipoderoxidación en glándula submandibular de ratas / Effect of carboplatin on malondialdehyde as a marker of lipoderoxidation in the submandibular gland of rats

El objetivo del presente trabajo fue evaluar el efecto de carboplatino (Cp) en homogenato de glándula submandibular de ratas Wistar a través de la determinación de los niveles de malondialdehido (MDA), como principal producto final de la lipoperoxidación, en un modelo experimental. Se utilizaron 16 ratas Wistar macho de tres meses de edad, alojadas en jaulas individuales, con temperatura e iluminación controlada y dieta libre. Se utilizó un diseño completamente aleatorizado y se establecieron dos grupos experimentales: 1) Control (C), administrándose una dosis intraperitoneal de solución salina durante un día, n: 8, 2) Animales tratados con carboplatino (Cp) aplicándose una dosis i.p. de 100 mg/Kg de peso corporal durante un día, n: 8. Los animales fueron ayunados por 24 horas y posteriormente anestesiados. Seguidamente se extirparon ambas glándulas submandibulares. Se analizaron los niveles de malondialdehido en homogenato de glándula submandibular en ambos grupos experimentales. Las variaciones entre los grupos analizados se evaluaron mediante prueba T de Student para muestras apareadas, fijando un p-valor <0,05 para significación estadística. Proyecto aprobado por CICUAL. Facultad de Ciencias Médicas (UNC). El grupo de Ratas C mostró una concentración de 7.32± 0.48µmol/mg de glándula. El grupo Cp tuvo una concentración de 12.57 ± 0,71 µmol/mg de glándula, expresando una disminución significativa respecto del grupo control p<0.0006. Cp en la dosis ensayada provocaría una disminución de la lipoperoxidación en glándula submandibular de ratas. Posiblemente la batería antioxidante glandular neutralizaría el estrés oxidativo de las células acinares. Estos resultados sugieren evaluar a futuro la actividad de superóxido dismutasa (SOD) y niveles de ácido úrico (AU)

Pehuén (Araucaria araucana) seed residues are a valuable source of natural antioxidants with nutraceutical, chemoprotective and metal corrosion-inhibiting properties.

In the last decade, green chemistry has been attracting great interest in many contexts, including, among others, natural antioxidants. However, only a few works deal with natural residue extracts and biowaste, which could be an efficient, economical and environmentally friendly source for the production of useful compounds. In this study, we look for antioxidant activity in Araucaria araucana seeds, an iconic pine species of the Argentine and Chilean Patagonia commonly known as "pehuén". Piñones are the edible pehuén seeds, and it is estimated that approximately 40 tons of piñones are harvested annually in Argentina and Chile. The chemical composition, antioxidants, metal corrosion-inhibiting properties and biological activity of edible and discarded piñón tissues were determined. Acute toxicity was discarded by in vitro testing and double fluorescent staining. Biological activity was evaluated in vivo by determining redox markers in salivary glands from rats treated with Cyclophosphamide (an oncological drug). All piñón tissues had antioxidants and antioxidant activity, with the coats showing the highest levels (up to 404 µg ascorbic acid equivalent per mg). The coats, in particular, had high gallic acid, catechin, quercetin and tannin contents, and more antioxidant activity, polyphenols and flavonoids than berries from the region. Results by X-ray fluorescence spectrometry showed that Na, Mg, P, S, Cl, K, and Ca were majority elements in the coat, embryo and endosperm. Furthermore, coat extract also showed significant anti-corrosion activity and in vivo protection against oxidative damage. The results indicate that piñón biowaste is a low-cost attractive source of natural antioxidants with potential nutraceutical, medical and metal corrosion protection applications.

5-fluorouracil and cyclophosphamide modify functional activity in submandibular gland of rats / El 5-fluorouracilo y la ciclofosfamida modifican la actividad funcional en la glándula submandibular de las ratas

Objective: Chemotherapy treatment against cancer produce systemic toxicities, among which are those related to important structures of the stomatognathic system and its functional activity. 5 Fluorouracil (5-FU) and cyclophosphamide (Cf) are drugs widely used in solid tumors and in bone marrow transplantation, respectively. The objective of this work was to evaluate the toxicity of these drugs regarding functional activity of the submandibular glands, by measuring the percentage of glycogen consumption in two experimental models. Material and Methods: 84 male Wistar rats aged three months were used, housed in individual cages, with controlled temperature and lighting and ad libitum diet. They were divided into four experimental groups: 1) Control (C); 2) Treated with 5-FU+leucovorin (LV) at 20 and 10mg/Kg of body weight respectively for five consecutive days; 3) treated with Cf i.p. at 50mg/Kg of body weight for two consecutive days; and 4) rats with paired feeding (PF): for five and two days respectively, the amount administered resulted from the average of the ingested food of groups 2 and 3. Both submandibular glands were excised. The submandibular glycogen concentration was analyzed at initial time (t0) and after 60 minutes of mechanical stimulation (t60). Results: the average variation changed significantly between time 0 and 60 in the groups C and PF. (p-value=0.0001), the 5-FU + LV treatment group had an average concentration higher at t0 than groups C and PF, without significant consumption at T60. While group Cf showed a lower average concentration at time 0 with respect to groups C and PF, without significant consumption at T60. Conclusion: 5-FU+LV and Cf affect the metabolism of carbohydrates, decreasing the use of glycogen as a metabolic substrate. In the present experimental model, the toxicity of these drugs affected the functional activity of the submandibular gland.

Objetivo: el tratamiento de quimioterapia contra el cáncer produce toxicidades sistémicas, entre las que se encuentran las relacionadas con estructuras importantes del sistema estomatognático y su actividad funcional. El 5-fluorouracilo (5-FU) y la ciclofosfamida (Cf ) son fármacos ampliamente utilizados en tumores sólidos y en trasplantes de médula ósea, respectivamente. El objetivo de este trabajo fue evaluar la toxicidad de estos fármacos con respecto a la actividad funcional de las glándulas submandibulares, midiendo el porcentaje de consumo de glucógeno en dos modelos experimentales. Material y Métodos: se utilizaron 84 ratas Wistar machos de tres meses de edad, alojadas en jaulas individuales, con temperatura e iluminación controladas y dieta ad libitum. Se dividieron en cuatro grupos experimentales: 1) Control (C); 2) Tratados con 5-FU+leucovorina (LV) a 20 y 10mg/Kg de peso corporal, respectivamente, durante cinco días consecutivos; 3) tratados con Cf i.p. a 50mg/Kg de peso corporal durante dos días consecutivos; y 4) ratas con alimentación por parejas (PF): durante cinco y dos días respectivamente, la cantidad administrada resultó del promedio de los alimentos ingeridos de los grupos 2 y 3. Ambas glándulas submandibulares fueron extirpadas. La concentración de glucógeno submandibular se analizó en el momento inicial (t0) y después de 60 minutos de estimulación mecánica (t60). Resultados: la variación promedio cambió significativamente entre el tiempo 0 y 60 en los grupos C y PF. (p=0,0001), el grupo de tratamiento 5-FU+LV tuvo una concentración promedio más alta en t0 que los grupos C y PF, sin un consumo significativo en T60. Mientras que el grupo Cf mostró una concentración promedio más baja en el tiempo 0 con respecto a los grupos C y PF, sin un consumo significativo en T60. Conclusión: 5-FU + LV y Cf afectan el metabolismo de los carbohidratos, disminuyendo el uso de glucógeno como sustrato metabólico. En el presente modelo experimental, la toxicidad de estos medicamentos afectó la actividad funcional de la glándula submandibular.

Mucositis and salivary antioxidants in patients undergoing bone marrow transplantation (BMT)

OBJECTIVES: High doses of chemotherapy generate DNA damage in patients undergoing bone marrow transplantation (BMT), due to the production of reactive oxygen species (ROS). In order to evaluate the local defensive effectiveness of the patient undergoing BMT, the concentrations of the antioxidants superoxide dismutase (SOD) and uric acid (UA) were measured in saliva. STUDY DESIGN: Basal saliva samples were collected from 20 patients undergoing BMT at the Oncology Department, Sanatorio Allende (Córdoba), in the stages: initial, prior to conditioning therapy (I); middle: 7 to 10 days after BMT (M) and final stage, 30 days after discharge from isolation (F). SOD levels were determined using a RANDOX kit (RANSOD superoxide dismutase manual), and for uric acid enzymatic UOD / PAP spectrophotometric method, ( Trinder Color Kit , Wiener Lab) was used.RESULTS:85% of the patients developed oral mucositis. SOD concentration in the M stage was significantly higher (p<0.01) compared with stage I, and it reversed in stage F. UA concentration was significantly lower (p<0.001) in stage M compared with stage I, and in stage F it recovered the initial values. CONCLUSIONS: SOD increase in stage M coincided with the appearance of mucositis, which could be interpreted as a defensive mechanism of saliva against oxidative stress produced by chemotherapy. UA decrease in stage M would favour the development of higher degrees of mucositis

Mucositis and salivary antioxidants in patients undergoing bone marrow transplantation (BMT).

OBJECTIVES: High doses of chemotherapy generate DNA damage in patients undergoing bone marrow transplantation (BMT), due to the production of reactive oxygen species (ROS). In order to evaluate the local defensive effectiveness of the patient undergoing BMT, the concentrations of the antioxidants superoxide dismutase (SOD) and uric acid (UA) were measured in saliva. STUDY DESIGN: Basal saliva samples were collected from 20 patients undergoing BMT at the Oncology Department, Sanatorio Allende (Córdoba), in the stages: initial, prior to conditioning therapy (I); middle: 7 to 10 days after BMT (M) and final stage, 30 days after discharge from isolation (F). SOD levels were determined using a RANDOX kit (RANSOD superoxide dismutase manual), and for uric acid enzymatic UOD / PAP spectrophotometric method, ( Trinder Color Kit , Wiener Lab) was used. RESULTS: 85% of the patients developed oral mucositis. SOD concentration in the M stage was significantly higher (p<0.01) compared with stage I, and it reversed in stage F. UA concentration was significantly lower (p<0.001) in stage M compared with stage I, and in stage F it recovered the initial values. CONCLUSIONS: SOD increase in stage M coincided with the appearance of mucositis, which could be interpreted as a defensive mechanism of saliva against oxidative stress produced by chemotherapy. UA decrease in stage M would favour the development of higher degrees of mucositis.

Analysis of saliva samples from oncological patients treated with 5-fluorouracil and leucovorin calcium by scanning electron microscopy with energy dispersive system.

This work presents a chemical and morphological analysis of samples of saliva taken from patients who were under treatment with intravenous chemotherapy with 5-fluorouracil and leucovorin calcium. Samples of saliva were extracted from fifteen patients during the three stages of the treatment: The initial stage (previous to the chemotherapy), the intermediate stage (during the chemotherapy), and the final stage (twenty-one days after finishing the treatment). An amount of 50 µl was collected in each visit. Chemical contrast images were taken by means of scanning electron microscopy, and X-ray characteristic spectra were obtained from all the studied samples by using an energy dispersive system from all the studied samples. Images that correspond to the intermediate stage showed important differences with respect to the initial and final stages. In addition, X-ray spectra provided information about the present elements in saliva and their relative abundance allowed us to determine variations in the chemical composition. The backscattered electron images and X-ray spectra from the intermediate stage showed clusters of crystals with fluorine content higher than those obtained in initial and final stages. This fact probably indicates the passage of metabolites of 5-fluorouracil and leucovorin calcium from the plasma to the oral cavity. This finding enhances the hypothesis proposed by other authors about the secondary effects of the drugs on the stomatognathic system such as oral mucositis, dysgeusia, and xerostomia with or without hyposalivation.

Functional impairment in submandibular gland of rats induced by 5-fluorouracil and calcium leucovorin.

One of the main clinical problems during chemotherapy is the occurrence of severe systemic toxicities, including those related to the stomatognathic system, which contribute to reducing the patient's quality of life. The most frequent oral complications are mucositis, dysgeusia, inflammation, gingival bleeding and decreased salivary flow or hyposalivation, a factor that predisposes to xerostomia, and other local complications that alter the homeostasis of the system. The purpose of this study was to evaluate the functional activity of salivary glands in Wistar rats subject to chemotherapy by measuring salivary flow, glycogen levels and glandular tissue response to autonomic nervous system agonists. Five experimental groups were used: 1) Control group fed "ad libitum"; 2) 5-fluorouracil (20 mg/kg body weight); 3) Calcium leucovorin (10 mg/kg body weight); 4) 5-fluorouracil + calcium leucovorin (20 and 10 mg/kg, respectively) by intraperitoneal injection for five consecutive days and 5) control with paired diet. Groups 1 and 5 did not receive drugs. Treatment with fluorouracil + leucovorin produced an increase in stimulated salivary flow and a higher response to increasing doses of beta agonists compared to other experimental groups. In both groups treated with cytostatic drugs, blocking of glycogen consumption at the end of the experimental period was observed. Our work suggests that salivary secretion may be affected by a dual mechanism: the first would be toxicity induced by 5-FU, which would cause depression of the process of glucose utilization. The second mechanism would affect the sympathetic autonomic reflex arc. In this instance, the synergistic action of 5-FU + LV would have a negative effect on the nerve activity with a reduction of salivary secretion. This would explain the hyposalivation, cited by several authors in patients undergoing the 5-FU + LV scheme in the treatment of colon carcinoma.

Functional impairment in submandibular gland of rats induced by 5-fluorouracil and calcium leucovorin

One of the main clinical problems during chemotherapy is the occurrence of severe systemic toxicities, including those related to the stomatognathic system, which contribute to reducing the patient's quality of life. The most frequent oral complications are mucositis, dysgeusia, inflammation, gingival bleeding and decreased salivary flow or hyposalivation, a factor that predisposes to xerostomia, and other local complications that alter the homeostasis of the system. The purpose of this study was to evaluate the functional activity of salivary glands in Wistar rats subject to chemotherapy by measuring salivary flow, glycogen levels and glandular tissue response to autonomic nervous system agonists. Five experimental groups were used: 1) Control group fed "ad libitum"; 2) 5-fluorouracil (20mg/ kg body weight); 3) Calcium leucovorin (10 mg/kg body weight); 4) 5-fluorouracil + calcium leucovorin (20 and 10 mg / kg, respectively) by intraperitoneal injection for five consecutive days and 5) control with paired diet. Groups 1 and 5 did not receive drugs. Treatment with fluorouracil + leucovorin produced an increase in stimulated salivary flow and a higher response to increasing doses of beta agonists compared to other experimental groups. In both groups treated with cytostatic drugs, blocking of glycogen consumption at the end of the experimental period was observed. Our work suggests that salivary secretion may be affected by a dual mechanism: the first would be toxicity induced by 5-FU, which would cause depression of the process of glucose utilization. The second mechanism would affect the sympathetic autonomic reflex arc. In this instance, the synergistic action of 5-FU + LV would have a negative effect on the nerve activity with a reduction of salivary secretion. This would explain the hyposalivation, cited by several authors in patients undergoing the 5-FU+LV scheme in the treatment of colon carcinoma.

Uno de los principales problemas clinicos durante la quimioterapia es la aparicion de graves efectos toxicos sistemicos, incluidos los relacionados con el sistema estomatognatico, que contribuyen a la reduccion de la calidad de vida del paciente. Las complicaciones orales mas frecuentes son la mucositis, disgeusia, inflamacion, sangrado gingival y la disminucion del flujo salival o hiposalivacion, un factor que predispone a la xerostomia, y otras complicaciones locales que alteran la homeostasis del sistema. El objetivo de este estudio fue evaluar la actividad funcional de las glandulas salivales de ratas Wistar sometidas a quimioterapia, a traves de la medicion del flujo salival, los niveles de glucogeno y la respuesta del tejido glandular a agonistas del sistema nervioso autonomo. Se utilizaron cinco grupos experimentales: 1) Control con alimentacion "ad libitum"; 2) 5 - fluorouracilo (20 mg / kg de peso corporal); 3) Leucovorina calcica (10 mg/ kg de peso corporal); 4) 5 - fluorouracilo + leucovorina calcica (20 y 10 mg / kg, respectivamente) por via intraperitoneal durante cinco dias consecutivos, y 5) control con dieta apareada. Grupos 1 y 5 no recibieron drogas. El tratamiento con 5 - fluorouracilo + leucovorina produjo un aumento de flujo salival estimulado y una mayor respuesta a dosis crecientes de agonistas beta en comparacion con otros grupos experimentales. En ambos grupos tratados con citostaticos, se observo bloqueo del consumo de glucogeno al final del periodo. Nuestro trabajo sugiere que la secrecion salival puede estar afectada por un doble mecanismo: el primero seria la toxicidad inducida por 5-FU que causaria depresion del proceso de utilizacion de la glucosa. El segundo mecanismo afectaria el arco reflejo autonomo simpatico. En este caso, la accion sinergica de ambos farmacos de 5-FU + LV repercutiria negativamente sobre la actividad nerviosa con una reduccion de la secrecion salival. Esto explicaria la hiposalivacion citada por varios autores en pacientes sometidos al esquema 5-FU + LV en el tratamiento del carcinoma colorrectal.