Intestinal parasitic infections and risk factors among the population in Cambodia

@#Many species of helminths and protozoa caused intestinal parasitic infections (IPIs). It belongs to neglected tropical diseases (NTDs) and remains a major public health problem in several Southeast Asian countries. The present study aimed to investigate the prevalence of IPIs and associated risk factors among the population in Kratie Province in northeastern Cambodia and Phnom Penh is the capital that locates in southern Cambodia. Fecal specimens (n=366) were collected in 10 villages in Kratie Province and Phnom Penh from 2019 to 2021. They were processed using the formalin ethyl-acetate concentration technique (FECT) to investigate parasites at egg and cyst stages and then examined under a light microscope. The results revealed that the prevalence of IPIs among the population in Kratie Province (n=317) and Phnom Penh (n=49) was 16.12% (n=59); of Kratie Province (n=50, 13.66%) and Phnom Penh (n=9, 2.46%), 12.02% (n=44) were helminths and 4.10% (n=15) were protozoa. The parasitic infection rate was higher in males (9.02%) than in females (7.10%) and more likely to be due to helminths (7.38%) than protozoa (1.64%). Prevalence of Opisthorchis viverrini was the highest (5.74%), followed by those of Entamoeba coli (4.10%), hookworm (3.83%), Ascaris lumbricoides (1.10%), Hymenolepis nana (1.09%), Taenia spp. (0.54%), Trichuris trichiura (0.55%), and Enterobius vermicularis (0.27%), respectively. Moreover, O. viverrini infection was the most common infection in the >20-year age group in Kratie Province. In addition, the bivariate and multivariate analyses showed that the association between gender. Gender was a significant risk factor positively associated with O. viverrini and hookworm infections (ORadj=0.318, 95% CI=0.122-0.8270, P=0.019 and ORadj=0.085, 95% CI=0.017-0.436, P=0.003, respectively). In conclusion, the IPIs were highly prevalent, especially O. viverrini and hookworm infections, among the population in Cambodia. These IPIs impact the public health burden but can be prevented by education regarding good sanitary practices in this community.

Practical tips to using formalin-fixed paraffin-embedded tissue archives for molecular diagnostics in a South African setting

Background: Formalin-fixed paraffin-embedded (FFPE) tissue archives in hospitals, biobanks, and others offer a vast collection of extensive, readily available specimens for molecular testing. Unfortunately, the use of tissue samples for molecular diagnostic applications is challenging; thus, the forensic pathology FFPE tissue archives in Africa have been a largely unexploited genetic resource, with the usability of DNA obtainable from these samples being unknown.Intervention: The study, conducted from January 2015 to August 2016, determined the usefulness of FFPE tissue as a reliable source of genetic material for successful post-mortem molecular applications and diagnostics. Formalin-fixed paraffin-embedded tissue samples were collected and archived from autopsies conducted over 13 years in the forensic medicine department of the University of Pretoria (Pretoria, South Africa). Deoxyribonucleic acid from FFPE tissue samples and control blood samples was amplified by high-resolution melt real-time polymerase chain reaction before sequencing. The procurement parameters and fixation times were compared with the quantity and quality of the extracted DNA and the efficiency of its subsequent molecular applications.Lessons learnt: This study has shown that FFPE samples are still usable in molecular forensics, despite inadequate sample preparation, and offer immense value to forensic molecular diagnostics.Recommendations: FFPE samples fixed in formalin for more than 24 h should still be used in molecular diagnostics or research, as long as the primer design targets amplicons not exceeding 300 base pairs.

Essential Oil from the Stem Bark of Casuarina equisetifolia Exerts Anti-inflammatory and Anti-nociceptive Activities in Rats

Abstract Herein the chemical constituents and the anti-pain properties of the essential oil from the stem bark of Casuarina equisetifolia L. (Casuarinaceae) grown in Nigeria were evaluated. The essential oil was obtained by hydrodistillation method in an all glass Clevenger-type apparatus, and characterized by gas chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS). The hot plate method was used to determine the anti-nociceptive property whereas the anti-inflammatory activity was evaluated by carrageenan-induced and formalin experimental models. The pale-yellow essential oil was obtained in yield of 0.21% (v/w), calculated on a dry weight basis. The main constituents of the essential oil were methyl salicylate (30.4%), a-zingiberene (15.5%), (E)-anethole (9.5%), b-bisabolene (8.6%), b- sesquiphellandrene (6.9%), and ar-curcumene (6.2%). In the anti-nociceptive study, the rate of inhibition increases as the doses of essential oil increases with optimum activity at the 30th and 60th min for all tested doses. The essential oil displayed anti-nociceptive activity independently of reaction time at the highest tested dose (200 mg/kg). The essential oil of C. equisetifolia moderately reduced pain responses in early and late phases of the formalin test. The oil inhibited the paw licking in the neurogenic phase (60-63%) compared to the late phase of the formalin test. The carrageenan- induced oedema model revealed the suppression of inflammatory mediators within the 1st - 3rd h. Thus, C. equisetifolia essential oil displayed both anti-nociceptive and anti-inflammatory activities independent of the dose tested. The anti-inflammatory and anti-nociceptive activities of C. equisetifolia essential oil are herein reported for the first time

Alcohol-based fixatives can better preserve tissue morphology than formalin / Fijadores a base de alcohol pueden preservar mejor la morfología tisular que la formalina

SUMMARY: Fixation is a crucial step in processing of tissue specimen for preservation of cellular architecture and composition of cells. Alcohol-based fixatives are considered some of the most promising alternatives to formalin. We evaluated the performance of alcohol-based fixatives (EthMeth and methacarn) and formalin as a comparator fixative in the research laboratory. Following 24 hours of fixation, tissue morphology and cellular details of the liver, spleen and brain (cerebral cortex) were evaluated. Morphological characteristics were evaluated by gross observations and analyzing cellular details, tissue architecture and overall staining characteristics (Hematoxylin and Eosin). EthMeth and methacarn fixation gave generally comparable and satisfactory results on the tissue morphology and subsequent identification of tissue characteristics. Particularly, tissues were well preserved and all nuclear as well as cytoplasmic details were clearly visible. However, formalin fixed tissues showed some peculiarity such as improper fixation, mild shrinkage, and alterations of tissue components. These results confirm that alcohol-based fixation is the superior alternative to formalin for preservation of tissue morphology. However, it is required to standardize the formalin-free methods and harmonize diagnosis in the laboratory worldwide.

RESUMEN: La fijación es un paso crucial en el procesamiento de muestras de tejido para preservar la arquitectura celular y la composición de las células. Los fijadores a base de alcohol se consideran algunas de las alternativas más prometedoras a la formalina. Evaluamos el rendimiento de los fijadores a base de alcohol (EthMeth y metacarn) y formalina como fijador comparativo en el laboratorio de investigación. Después de 24 horas de fijación, se observó la morfología del tejido y los detalles celulares del hígado, bazo y corteza cerebral. Se evaluaron las características morfológicas mediante observaciones generales y analizando detalles celulares, arquitectura de tejidos y características generales de tinción (hematoxilina y eosina). La fijación de EthMeth y metacarn dio resultados generalmente comparables y satisfactorios en la morfología del tejido y la posterior identificación de las características del mismo. Particularmente, los tejidos estaban bien conservados y todos los detalles nucleares y citoplasmáticos eran claramente visibles. Sin embargo, los tejidos fijados con formalina mostraron cierta peculiaridad, tal como una fijación inadecuada, la contracción leve y alteraciones de los componentes del tejido. Estos resultados confirman que la fijación a base de alcohol es la mejor alternativa a la formalina, para preservar la morfología del tejido. Sin embargo, es necesario estandarizar los métodos sin formalina y armonizar el diagnóstico en los laboratorios.

Analgesic and neuromotor effects of two extracts of Zanthoxylum rhoifolium (Rutaceae) in the Swiss albino mouse (Mus musculus) / Efectos analgésicos y neuromotores de dos extractos de Zanthoxylum rhoifolium (Rutaceae) en el ratón albino suizo (Mus musculus)

Background: Colombia has a high diversity of medicinal plants, including Tachuelo (Zanthoxylum rhoifolium), a tree belonging to the family Rutaceae, which has been attributed an analgesic use by communities of the Colombian southwest. Nevertheless, this species has been scarce studied in Colombia. The studies have been limited to the isolation of compounds with antifungal and insecticide activity, using the leaves, branches, and bark of the plant in aqueous, and organic extracts. Neither pharmacological tests have been carried out, nor the analgesic and neuromotor activity have been tested from preparations with this plant's fruits. Objectives: To evaluate the analgesic and neuromotor effect of the aqueous and hexane extract of the fruits of Z. rhoifolium.Methods: As there were no preliminary reports of this study, the first observation was made through Irwin's test. The formalin and the rotarod test were performed to determine the analgesic and motor coordination effects, respectively. Results: All the evaluated treatments demonstrated to generate analgesia, anesthesia, passivity, reduction in the alarm reaction, and have antinociceptive activity in the formalin test; while only the high dose of the nonpolar extract generated a deficit in the motor performance of the rodents in the rotarod test. Conclusions: the antinociceptive effect of the aqueous and hexane extracts of this species' fruits was demonstrated. The hexane extract generated a neuromotor effect, which validates the ethnobotanical reports on Z. rhoifolium fruits.

Antecedentes: Colombia tiene una alta diversidad de plantas medicinales, incluyendo al Ta chuelo (Zanthoxylum rhoifolium), un árbol de la familia Rutaceae al que se le ha atribuido un potencial analgésico debido al uso por parte de las comunidades del Suroccidente Colombiano. Sin embargo, esta especie ha sido poco estudiada en Colombia, limitándose al aislamiento de compuestos con actividad antifúngica e insecticida, empleando las hojas, ramas y corteza de la planta en extractos acuosos y orgánicos; sin embargo, no se han realizado ensayos farmacológicos ni se han probado la actividad analgésica y neuromotora a partir de preparaciones con los frutos de esta planta. Objetivos: Evaluar el efecto analgésico y neuromotor del extracto acuoso y en hexano de los frutos de Z. rhoifolium. Métodos: Al no haber reportes preliminares de este estudio, se realizó la primera observación mediante el test de Irwin. Para determinar el efecto analgésico y neuromotor se realizó el test de formalina y el test de rotarod, respectivamente. Resultados: Todos los tratamientos evaluados demostraron generar analgesia, anestesia, pasividad, reducción en la reacción de alarma, tener actividad antinociceptiva en el test de formalina, mientras que solo la dosis alta del extracto apolar generó un déficit en el rendimiento motor de los roedores en el test de rotarod. Conclusiones: Se demostró el efecto antinociceptivo del extracto acuoso y en hexano de los frutos de esta especie y el efecto neuromotor generado por el extracto en hexano, validando los reportes etnobotánicos sobre los frutos de Z. rhoifolium.

Comparison of Three Parasitological Stool Examination Methods with the Formalin-Ethyl Acetate Procedure for the Diagnosis of Intestinal Parasites in Humans

Aims:The objective was to compare the sedimentation spontaneous in tube technique(SSTT), the direct smear, the zinc sulfhate(surface film and sediment analyzed) methods with the formalin-ethyl acetate method as the reference standard.Study Design:This a cross-sectional study performed in two populations aroundChapala lake, Jalisco, Mexico.Place and Duration of Study:Sample Department of Medical Sciences and Life, University Center of the Ciénega, University of Guadalajara, Ocotlán, Jalisco, Mexico. Department of Physiology, Health Sciences University Center. University of Guadalajara, Jalisco. México, between June 2018 andJuly 2019.Methodology:Sample: We included a total of 297 samples and were analyzed by the direct smear, the zinc sulfhate (surface film and sediment analyzed), the SSTT and formalin-ethyl acetate methods.Results:The SSTT was able to detect 40% of intestinal parasites, very good agreement for Entamoebacomplex, Entamoeba coli, andAscarislumbricoideskappa index=0.697, 0.791, 0.696, respectively and excellent agreement for Giardia lambliakappa index = 0.843. Regarding the isolation ofBlastocystisspp, only a poor agreement was found among all techniques. The SSTT was able to detect multiple parasites with a sensitivity of 82.90% and specificity 83.80 kappa index= 0.649.Conclusion:The SSTT showed a very good agreement for the diagnosis intestinal polyparasitism which could represent another alternative for the concentration and identification protozoans and intestinal helminthes in low-resource settings

Efecto de gas de formalina sobre el recuento de mesófilos en nailon comercial destinado a procedimientos quirúrgicos / Effect of formaline gas on the counting of mesophiles in commercial nylon destined to surgical procedures

RESUMEN La investigación se realizó a fin de evaluar el efecto del gas de formalina sobre el recuento de mesófilos en nailon comercial (poliamida) destinado a procedimientos quirúrgicos. En primer lugar, se evaluó la contaminación de una muestra del material comercial de 1 g expuesta al ambiente de quirófano por 72 horas, a través de la técnica de recuento en placa para mesófilos; se obtuvieron 850 UFC/g. Una vez comprobada la contaminación de la poliamida comercial, esta se sometió a gases de formalina en comprimidos. Se colocaron 5 muestras (n=5) de nailon de 1 g cada una en 5 recipientes herméticos de 1 litro con 1 gramo de formalina cada uno; estos recipientes se almacenaron en un mueble en sala de esterilización a un metro de altura del piso y posteriormente, fueron abiertos y cultivados a través de la técnica de recuento en placa para mesófilos, uno por día a lo largo de 5 días a intervalos de 24 horas. Los resultados obtenidos no registraron crecimientos de microorganismos a partir de las 24 horas y durante los 5 días posteriores y se encontraron diferencias estadísticamente significativas (p < 0,05). Se concluye que bajo las condiciones del presente estudio el tiempo de esterilización de la formalina sobre nailon comercial es de 24 horas.

ABSTRACT Hie present research work was carried out with the purpose of evaluating the effect of formalin gas on the count of mesophiles in commercial nylon (polyamide) destined to surgical procedures. Firstly, the contamination of a 1 g commercial nylon sample, exposed to the operating room environment for 72 hours, was evaluated through the plate counting technique for mesophiles; it yielded a result of 850 CFU / gram. Once the contamination of the commercial polyamide was checked, it was subjected to formalin gases in tablets. Five nylon samples (n=5) of 1 gram each were placed in 5 1 liter airtight containers containing 1 gram of formalin; Hese containers were stored in a cabinet in the sterilization room one meter above the floor, and later opened and cultivated through the plate counting technique for mesophiles, one per day for 5 days at 24 hour intervals. The results obtained after being exposed to formalin gases in tablets did not register growths of microorganisms after 24 hours and during the 5 days after the study, finding statistically significant differences (p < 0.05) in relation to the studied times concluding under the conditions of the present study that the sterilization time of the formalin on commercial nylon is equal to 24 hours.

Unintended Formalin Injection instead of Local Anesthetic Agent: Its Causes & Management

Mistakes do happen in every profession instead of properprecautions. Mishaps are those unfortunate accidents thatoccur during treatment, some owing to inattention to detail,others being very unpredictable and some are due to thenegligence of the employee. Dental negligence may lead tosevere complications and may threaten one's life. This casereport presents an unconventional case of administeringformalin in lieu of local anesthesia in a dental clinic leading tochemical cellulitis and its management.

In vivo methods for the evaluation of anti-inflammatory and antinoceptive potential / Métodos in vivo para avaliação do potencial anti-inflamatório e antinociceptivo

ABSTRACT BACKGROUND AND OBJECTIVES: The constant search for bioactive compounds with anti-inflammatory and antinociceptive activities are of interest to research centers. For the characterization of these activities, trials on guinea pigs are necessary. Therefore, the purpose of this study was to demonstrate some methods to evaluate the anti-inflammatory and antinociceptive potential of natural products. CONTENTS: A stimulus is required to evaluate these activities, and the induction of inflammatory or nociceptive process can be by chemical inducers like formaldehyde, carrageenan, among others, or electronic equipment such as the hot plate. For all assays, the baseline and post-dose measurement of the studied compound is always compared with a control group. The planning of the experiment, as well as its conduct in accordance with well-established protocols, are important tools in the success of the work. The tests presented evaluated the antinociceptive and anti-inflammatory activity as well as the mechanisms involved. CONCLUSION: It was possible to evaluate that the tests present in the literature today meet the researcher's need for the elucidation of the anti-inflammatory and antinociceptive activity of new compounds.

RESUMO JUSTIFICATIVA E OBJETIVOS: A busca constante por compostos bioativos com atividade anti-inflamatória e antinociceptiva são de interesse dos centros de pesquisas. Para a caracterização dessas atividades são necessários ensaios em cobaias. Frente a isso, o objetivo deste estudo foi demonstrar alguns métodos para a avaliação do potencial anti-inflamatório e antinociceptivo de produtos naturais. CONTEÚDO: Para a avaliação dessas atividades é necessário um estímulo, sendo que a indução de processo inflamatório ou nociceptivo pode ser por indutores químicos como formol, carragenina, entre outros, ou ainda, equipamentos eletrônicos como placa quente. Para todos os ensaios, sempre é realizada a mensuração basal e posterior à administração do composto que está sendo estudado em comparação com um grupo controle. O planejamento do experimento, assim como toda a condução conforme protocolos já bem ilustrados, são ferramentas importantes no êxito do trabalho. Os testes apresentados avaliaram atividade antinociceptiva e anti-inflamatória assim como mecanismos envolvidos. CONCLUSÃO: Foi possível avaliar que os testes presentes na literatura hoje, atendem a necessidade do pesquisador na elucidação da atividade anti-inflamatória e atividade antinociceptiva de novos compostos.

A valepotriate-enriched fraction from Valeriana glechomifolia Meyer inhibits leukocytes migration and nociception in formalin test in rodents

Abstract Previous pre-clinical studies demonstrated that a valepotriates enriched fraction from Valeriana glechomifolia F.G. Mey., Caprifoliaceae, was effective against lipopolysaccharide from Escherichia coli (LPS)-induced sickness behavior as well as significantly decreased the cortical expression of pro inflammatory cytokines interleukin-1β and tumor necrosis factor-α. Other studies revealed anti-inflammatory properties of V. wallichii and V. amurensis. These findings open up new perspectives for Valeriana genus pharmacology, once it has been commonly associated to sedative and anxiolytic properties. The aim of this study was to investigate the antichemotactic, antinociptive and anti-inflammatory activities of a valepotriate-enriched fraction obtained from aerial and subterranean parts of V. glechomifolia submitted to supercritical CO2 extraction. The biological activities were assessed by means of formalin test in CF1 mice and Wistar rat's leukocytes migration assay (modified Boyden chamber method). Valepotriate-enriched fraction (1, 10 and 30 mg/kg, p.o.) inhibited the nociceptive behavior in the late phase of the formalin test in a dose dependent manner. The effect of the valepotriate-enriched fraction highest dose was comparable with that of diclofenac 50 mg/kg (p.o.). Valepotriate-enriched fraction (0.1-1 µg/ml) inhibited the leukocyte migration induced by lipopolysaccharide from Escherichia coli in a concentration dependent manner. This antichemotatic effect was comparable with that of indomethacin (0.1-1 µg/ml) and better than diclofenac (1 µg/ml) effect. This study demonstrated for the first time that a valepotriate-enriched fraction obtained from V. glechomifolia display a peripheral anti-inflammatory like activity.

Preparación de hemisferios cerebrales para disección de tractos / Preparation of cerebral hemispheres for tracts dissection

Desde la antigüedad se han desarrollado técnicas para el estudio del cerebro con fines didácticos o neuroquirúrgicos. Hacia 1934 Josef Klingler desarrolla una técnica de preparación de hemisferios cerebrales que basada en la fijación con formalina y el congelamiento para aislar los tractos cerebrales. El objetivo de la presente artículo ha sido analizar los métodos de preparación utilizados para la disección de tractos en cerebros humanos y de animales. Se realizó una revisión de la literatura en las bases de datos Web of Science, Scopus, Pubmed, Medline y Scielo, utilizando como descriptores: Disección, Cerebro, Tracto, con el operador booleano "AND" entre ellos, en los idiomas inglés y español, hasta junio de 2018. Fueron seleccionados 26 documentos, para el análisis se determinaron las variables: espécimen, número de hemisferios cerebrales, concentración de formalina, tiempo de fijación, temperatura, tiempo de congelamiento y tractos identificados. En la literatura seleccionada, un total de 410 hemisferios cerebrales fueron analizados, 372 de humanos y 38 de animales; 330 fueron conservados en formalina al 10 %, 20 en formalina al 5 % y el resto en otras concentraciones. El tiempo de fijación fue variable entre 10 y 180 días, así como la temperatura y tiempo de congelación (-10 ºC y -20 ºC, entre 8 y 30 días). En todos los casos se reportó que, en su totalidad o parcialmente, los fascículos cerebrales de asociación fueron aislados. En la preparación de hemisferios cerebrales para disección de tractos, Ludwig & Klingler (1956) recomiendan que en la fijación de los especímenes se utilice formalina al 5 %, sin embargo, el 80 % de los hemisferios utilizados fueron fijados en formalina al 10%, y en esta concentración, el tiempo de fijación, temperatura y tiempo de congelación fue variable, lográndose, en todos los casos analizados, la disección parcial o total de los tractos.

Since ancient times, techniques for the study of the brain have been developed for didactic or neurosurgical purposes. By 1934, Josef Klingler developed a cerebral hemisphere preparation technique based on formalin fixation and freezing to isolate the cerebral tracts. The aim of this article was to analyze the preparation methods used for tracts dissection in human and animal brains. A review of the literature using Web of Science, Scopus, Pubmed, Medline and Scielo databases, with the following descriptors: Dissection, Brain, Tract, with the boolean operator "AND" among them, also in spanish, until June 2018. Twenty-six documents were selected, and we analized the following variables: specimen, number of cerebral hemispheres, formalin concentration, fixing time, temperature, freezing time and tracts. In the selected literature, a total of 410 cerebral hemispheres were analyzed, 372 from humans and 38 from animals; 330 were preserved in 10 % formalin, 20 in 5 % formalin and the rest in other concentrations. The fixation time was variable between 10 and 180 days, as well as the temperature and freezing time (-10 ºC and -20 ºC, between 8 and 30 days). In all cases it was reported that, in whole or in part, the cerebral fascicles of association were isolated. In the preparation of cerebral hemispheres for dissection of tracts, Klingler recommend that 5 % formalin for the fixation of specimens; however, 80 % of the hemispheres used were fixed in 10 % formalin, and in this concentration, the time of fixation, temperature and time of freezing was variable, achieving, in all the cases analyzed, the partial or total dissection of the tracts.

Formalin Induced Nuclear Abnormalities In Relation With Duration Of Exposure: A Study Of Formalin Exposed Anatomists, Attenders And Students

Background: Exposure of Genotoxic agents like formalin causes cancer. The duration of exposure and thequantity of exposure plays an important role in inducing the nuclear abnormalities. The nuclear abnormalitiesare BN, KR, KL.Objective: This study provides an easy and inexpensive way to assess the formalin induced nuclear abnormalitiesin different groups with different duration of exposure.Materials and methods: Fifty individuals with different duration of exposure were included in our descriptivestudy. Oral squamous cells were collected with the wooden spatula. The slides were fixed with methanolglutaraldehyde fixative and stained with Giemsa, May-Grunwald stains. After air drying, 1000 cells were screenedfor nuclear abnormalities like BN, KR, KL.Results: Nuclear abnormalities like KR, KL, BN were assessed. KL is more in less than 1-year exposure group.KR ismore in long term exposure groups. BN cells are seen 6 – 10 and 10 – 15 years exposure group.Conclusion: The genotoxic effect of formalin was confirmed by the increase in the nuclear abnormalities especiallyby the increase in the frequency of KR in tong term exposed group and KL in less than 1-year exposed group

Comparative evaluation of phasic and chemical antinociceptive action of a conventional and a novel anticonvulsants in experimental models of tail flick and formalin test

Background: Some antiepileptic drugs have been shown to be clinically efficacious in treatment of neuropathic pain and are being used by clinician.Methods: This study determined the analgesic effect of gabapentin (a conventional anticonvulsant) and levitiracetam (a novel anticonvulsant) in rats in different types of acute and chronic nociceptive test like tail flick and formalin test and compared its potency with a conventional non opioid analgesic diclofenac.Results: Per oral administration of gabapentin produced no any marked effect on early phase response of formalin test but significantly suppressed the late phase response while levitiracetam produced no any type of significant effect in both phases. In tail flick test gabapentin as well as levitiracetam produced no any significant analgesic effect while diclofenac produced significant reduction of pain in tail flick test as well as in both phases of formalin test.Conclusions: Thus, we have observed that gabapentin produced antinociception in chronic pain as second phase of formalin test reflects chronic inflammatory pain while levitiracetam did not produce any type of antinociceptive effect as it could not suppress the pain significantly in both tail flick and formalin test.

Four percent formalin application for the management of radiation proctitis in carcinoma cervix patients: An effective, safe, and economical practice

Context: Radiotherapy is a very effective treatment modality for pelvic malignancies such as carcinoma of the cervix. However, it is quite common for chronic radiation proctitis (CRP) to manifest after radical radiotherapy. CRP is a source of significant morbidity, and there is a lack of effective treatment modalities. There also exists a general lack of guidelines on management of CRP. Aims: To assess the benefit from 4% formalin application for the treatment of Grade >2 CRP among patients previously treated with radical radiotherapy for cervical carcinoma. Settings and Design: This retrospective descriptive study involved 29 eligible patients who were treated from November 2010 - November 2015 for CRP with 4% formalin application. Materials and Methods: Of the 1864 patients of carcinoma cervix treated during the said patients, 29 patients fulfilled the eligibility criteria. Eligible patients were invited telephonically for follow-up and were assessed for response and complications of the procedure. Results: The treatment of hemorrhagic radiation proctitis with local formalin instillation is effective, well tolerated and safe procedure. The procedure is inexpensive, technically simple and can be done on an outpatient basis. 62% patients had complete freedom from rectal bleed, while 34.5% patients had partial benefit. Only one patient required diversion colostomy for persistent bleeding.

A comparative study of Thiel soft-embalmed and formalin preserved cadavers for anatomy dissection

@#<p><strong>Background and Objective:</strong> Cadavers are the main teaching and learning tools utilized by faculty and students of Gross Anatomy. The conventional procedure of using formalin to preserve cadavers has its limited benefit due to its obnoxious odor, darkened color, indistinguishable small structures, rigid joints, with friable organs and tissues. Using the Thiel Method, cadavers are known to have: life-like presentation, a high degree of suppleness, natural mobilization of various body parts with tolerable odor, natural color, and antimicrobial and antifungal efficacy. The aim of this study was to compare the Thiel soft-embalmed cadaver and the formalin-preserved cadaver for use in Gross Anatomy dissection.</p><p><strong>Materials and Method:</strong> Six formalin-preserved cadavers were compared with six Thiel soft-embalmed ones and were evaluated by 160 first year medical students and 10 faculty members of Gross Anatomy from the University of the Philippines College of Medicine using a visual analogue scale that evaluated the cadaver's presenting color. A Likert scale was utilized to assess the skin and muscle texture, blood vessel integrity, odor, joint flexibility, and state of cadaver preservation. Their perceptions on the ease of the dissection process, distensibility, and differentiation of structures were also elicited through a questionnaire checklist.</p><p><strong>Results:</strong> Results show that the Thiel Method-preserved cadavers were rated higher compared to the formalinpreserved cadavers in terms of: 1) presenting color; 2) tolerable odor; 3) skin and muscle texture; 4) blood vessel integrity; 5) joint flexibility; and 6) state of cadaver preservation after 1 year. However, the ease of the dissection process, the distensibility of structures, and the differentiation of the various structures were easily managed in the formalin-preserved cadavers while the feasibility of practicing laparoscopy, endoscopy, and intubation were seen in the Thiel soft-embalmed cadavers. Results validate the claims of previous studies on the advantages presented in using the Thiel Method.</p><p><strong>Conclusion:</strong> Cadavers preserved utilizing the Thiel Method are life-like and can be used for Gross Anatomy dissection. The data gathered support the feasibility of using them also in research and training.</p>

Antinociceptive local activity of 4-allyl-1-hydroxy-2-methoxybenzene (eugenol) by the formalin test: an anti-inflammatory effect

Eugenol has been employed for decades as a condiment, an antimycotic, an antibacterial, an antiviral, and an antioxidant, and it is one of the natural analgesics most frequently utilized for pain and inflammation. Our objective was to determine the analgesic/anti-inflammatory effect of eugenol compared with diclofenac, naproxen, and tramadol using the formalin test. The formalin method was used in 6- to 10-week-old Wistar rats (weighing 250 g each) divided into six groups: saline (0.9%); formalin (5%); diclofenac (250 µg/kg); naproxen (400 µg/kg); tramadol (500 µg/kg), and eugenol (1,400 µg/kg), in the intraplantar part of the hind-end trunk of the rats, with n = 5 per group. Eugenol diminished 44.4% of nociceptive behavior in phase 1 and 48% in phase 2 (p ≤0.05 vs formalin). Eugenol was shown to be 1.14 times more effective than diclofenac, but 1.62 and 1.75 times less effective than naproxen and tramadol, respectively, in phase 1 and 1.45 times less effective than diclofenac and naproxen and 1.66 less effective than tramadol in phase 2 (p ≤0.05). These data suggest that eugenol possesses moderate activity in the acute pain phase and greater activity in inflammatory-type pain, and both effects are comparable to those produced by diclofenac and are less than the effects produced by naproxen and tramadol in the formalin test

DNA re-use after immunohistochemical staining of FFPE sections of non-small cell lung cancer / 医学研究生学报

Objective　The purpose of this study was to evaluate the quality of DNA from the formalin-fixed paraffin-embedded (FFPE) specimens of non-small cell lung cancer (NSCLC) after immunohistochemical staining and investigate DNA extraction by immunohistochemical staining of the specimens in small in number or difficult to obtain and the feasibility of related molecular tests.　Methods　We randomly collected 50 FFPE biopsy specimens of NSCLC in our Department of Pathology from June 2017 to December 2017 and sliced each into 12 sections, of which, 6 were directly subjected to DNA extraction (the control group) and the other 6 to immunohistochemical Envision two-step staining for DNA extraction (the experimental group). Then, we detected the mutations of the epidermal growth factor receptor (EGFR), kirsten rat sarcoma viral oncogene (KRAS) and V-raf murine sarcoma viral oncogene homolog B (BRAF) in all the DNAs extracted.　Results　No statistically significant differences were observed between the experimental and control groups in the DNA concentration and purity in the 50 FFPE biopsy specimens of NSCLC (P>0.05). Of the 50 NSCLC FFPE specimens of the experimental group, 20 (40%) showed the mutation of EGFR, 8 (16%) exhibited that of KRAS, and 5 (10%) manifested that of BRAF. In the other 50 specimens of the control group, 33 showed the mutations of EGFR, KRAS and BRAF. A 100% consistency was found in the results of detection between the experimental and control groups (P=0.000, Kappa=1.000).　Conclusion　 High-quality DNA can be extracted after immunohistochemical staining from NSCLC FFPE specimens, especially those small in number or difficult to obtain, and can be used for downstream molecular analysis of target genes, which is a good method for specimen recycling and provides a solution for subsequent molecular test of scarce or difficult-to-obtain clinical samples.

Multistaining Optimization for Epstein-Barr Virus–Encoded RNA In Situ Hybridization and Immunohistochemistry of Formalin-Fixed Paraffin-Embedded Tissues Using an Automated Immunostainer

BACKGROUND: Single staining is commonly performed for practical pathologic diagnoses. However, this method is limited in its ability to specify cellular morphology and immunophenotype and often requires consumption of limited tissue. This study aimed to describe an optimized protocol for multiple in situ hybridization (ISH) and immunohistochemistry (IHC). METHODS: The quality of multistaining was evaluated by carefully changing each step of ISH and IHC in an angioimmunoblastic T-cell lymphoma (AITL) case on a Ventana BenchMark XT automated immunostainer. The optimized protocols were also performed using another immunostainer and in 15 cases of five Epstein-Barr virus (EBV)–associated malignancies using formalin-fixed paraffin-embedded tissue. RESULTS: The quality of various ISH-IHC staining protocols was semi-quantitatively evaluated. The best EBV-encoded RNA (EBER)-ISH/double IHC staining quality, equivalent to single staining, was obtained using the following considerations: initial EBER-ISH application, use of protease and antigen retrieval reagent (cell conditioning 1 [CC1] treatment time was minimized due to impact on tissue quality), additional baking/deparaffinization not needed, and reduced dilution ratio and increased reaction time for primary antibody compared with single immunostaining. Furthermore, shorter second CC1 treatment time yielded better results. Multiple staining was the best quality in another immunostainer and for different types of EBV-associated malignancies when it was performed in the same manner as for the Ventana BenchMark XT as determined for AITL. CONCLUSIONS: EBER-ISH and double IHC could be easily used in clinical practice with currently available automated immunostainers and adjustment of reagent treatment time, dilution ratio, and antibody reaction time.

La inflamación inducida por formalina favorece el desarrollo del melanoma B16 en ratones Balb/C / inflamation induced by formaline promotes the development of melanoma B16 in Balb/C mice

El cáncer es la segunda causa de muerte en el mundo representando el melanoma 1% de todos los tipos de cáncer. Se ha planteado que el cáncer es una enfermedad inflamatoria sistémica que genera radicales libres causando mutaciones y liberan factores tróficos que favorecen la iniciación tumoral y la proliferación celular, respectivamente. Con el objetivo de estudiar el efecto de la inflamación inducida por formalina sobre el desarrollo del melanoma B16, 22 ratones Balb/C fueron distribuidos en tres grupos: Control Melanoma, Melanoma-Formalina y Control Formalina. A los grupos CF y MF se les aplicó 20 µl de Formalina al 2% en el dorso de la pata trasera derecha a nivel subcutáneo; a los grupos CM y MF se le trasplantaron 100.000 células melanocíticas vía subcutánea en la superficie plantar de la pata derecha, 24 horas posteriores a la formalina. Los ratones del grupo CF desarrollaron una inflamación que fue máxima entre la primera y segunda semana y luego cedió progresivamente hasta desaparecer a la sexta semana. Los ratones del grupo CM desarrollaron máculas tumorales hasta 30 mm² que involucionaron espontáneamente. Los ratones del grupo MF desarrollaron masas tumorales que alcanzaron hasta 300 mm³ entre la 3-4 semanas post-trasplante y luego disminuyeron progresivamente de volumen. Los ratones de los grupos CF y MF disminuyeron significativamente de peso respecto al grupo CM. En conclusión, la inflamación inducida por formalina favorece el desarrollo tumoral en un modelo alogénico de melanoma maligno.

Cancer is the second cause of death around the world, representing melanoma 1% of all cancer. It has been suggested that cancer is a systemic inflammatory disease that generates free radicals causing mutations and releasing trophic factors that favors tumor initiation and cell proliferation. In order to study the effect of formalin-induced inflammation on the development of B16 melanoma, 22 Balb/C mice were divided into three groups: Control Melanoma (CM), Melanoma-Formalin (MF) and Control Formalin (CF). CF and MF groups were injected with 20 µl of 2% formalin on the back of the right paw at the subcutaneous level; CM and MF groups were transplanted with 100,000 melanocytic cells subcutaneously in the plantar surface of the right paw, 24 hours after formalin. CF group mice developed an inflammation that was maximal between the first and second week, then progressively diminished until disappearance by the sixth week. CM group mice developed tumoral macules up to 30 mm², which involute spontaneously. MF group mice developed tumor masses that reached up to 300 mm³ between 3-4 weeks post-transplant and then progressively decreased in volume. CF and MF mice significantly decreased in weight with respect to CM group. In conclusion, inflammation induced by formalin favors tumor development in an allogenic model of malignant melanoma, indicating that anti-inflammatory treatments may be useful in the management of melanoma.

Preclinical screening of a novel compound, 2-chlorothiophene for analgesic activity in swiss albino mice

Background: Pain is often the first indication of disease or injury. Analgesics are the drugs used clinically for controlling pain. They relieve pain as a symptom, without affecting its cause. Currently available options are nonsteroidal anti-inflammatory drugs (NSAIDs) and opioid analgesics for the management of pain. Long term use of existing analgesics causes significant disturbances in the body system. A search for new, safe and cost effective analgesic compound is in progress. Hence a study on 2-chlorothiophene, a novel compound has been carried out in different experimental animal models.Methods: The central analgesic activity of 2-chlorothiophene was evaluated by eddy’s hot plate method and compared to standard central analgesic, morphine. Both central and peripheral analgesic activities of 2-chlorothiophene were evaluated by formalin induced paw licking in mice and compared to a standard drug, aspirin.Results: There were 40mg/kg dose of 2-chlorothiophene has shown maximum Pain Inhibition Percentage (PIP) of 46.15% at 60 min compared to 128% by morphine in eddy’s hot plate method. Under Formalin test, 20mg/kg dose of 2-chlorothiophene has shown maximum PIP of 22.91% in early phase and 52.63% in late phase compared to 12.5% and 47.37% by aspirin. The results were statistically significant with p<0.05.Conclusions: 2-chlorothiophene found to have minimal central analgesic activity and significant peripheral analgesic activity as evident in eddy’s hot plate and formalin tests.