The role and diagnostic value of the most common methods for diagnosing Helicobacter pylori infection

The purpose of the study was to identify and evaluate the practical value of the most common diagnostic methods for Helicobacter pylori infection in patients with chronic inflammation of the gastric mucosa. Materials and methods. The study involved 104 people. All the patients examined underwent esophagogastroduodenoscopy with a rapid urease histochemical test for Helicobacter pylori and a standard five-point biopsy of the gastric mucosa for morphological evaluation and bacterioscopy. If the result of the quick urease histochemical test coincided with the data of the biopsy study on Helicobacter pylori, no further examination was carried out. In case of discrepancy the patients additionally underwent the 13C-urease breath test. Results. It was found that the sensitivity of the rapid urease histochemical test in the area of the proposed model, according to our study, was 89.74% and the specificity was 46.15% when validating it using the additional methods. Similar calculations for the diagnostic method of staining by Giemsa showed sensitivity of 100% and specificity of 97.4%. The total number of the infected in the survey was 78 out of 104 people, which amounted to 75%. When analysing the severity of activity, inflammation and atrophic changes in the gastric mucosa, it was found that in the patients with two positive tests, the severity of the processes was significantly higher than in those with one positive test or all the tests were negative (p 0.05). When comparing the same processes in cases where only a quick urease histochemical test was positive and no signs of Helicobacteriosis were detected in any test, no significant differences were detected. Discussion. The rapid urease histochemical test showed good sensitivity of 89.74%, but unsatisfactory specificity of 46.15%, which severely limits its use. An unexpected result for this methodology was a large number of false positive tests, while the European recommendations indicate a more frequent occurrence of false negative results, which can be explained by differences in the production technology of the test systems. High sensitivity and specificity of the Helicobacter pylori test using Giemsa staining strongly depends on observing the methodology and experience of a specialist and cannot be recommended as a standard in the routine medical use. The study confirmed the recommendations for using at least two diagnostic tests to diagnose the infection, while the most common rapid urease histochemical test always needs confirmation. As a result of the analysis, it can be said that esophagastroduodenoscopy with a rapid urease histochemical test for Helicobacter pylori and standard diagnostic biopsy is the optimal method for diagnosing the pathology of the upper sections of the gastrointestinal tract. Moreover, 13C-UDT seems to be almost ideal as a screening technique and for evaluating treatment in cases where endoscopic monitoring is not necessary.

Anatomical Structure and Terpenoid Content of Zodia (Evodia suaveolens Scheff) Leaves

Zodia (Evodia suaveolens Scheff) is a member of Rutaceae contain terpenoids, triterpenoids, alkaloids, flavonoids, and xanthones which have anti-mosquito activity. This research aimed to observe the anatomical structure, the location, and distribution of terpenoid based on the leaves' age. Anatomical slides preparation of leaves were made using the paraffin embedding method with safranin staining. The distribution of terpenoid was analyzed by the histochemical assay. Leaf anatomical structure shows that the 3rd and 6th leaf bifacial (dorsiventral) consisted of the upper epidermis, mesophyll (palisade and sponge), collateral vascular bundle, parenchyma midrib, abaxial epidermis and oil glands in mesophyll that is underneath both epidermises. The diameter of oil glands with larger sizes was on the 6th leaf, whereas the density is not different in the 3rd and 6th leaves. The histochemical test showed that terpenoid was observed in the leaf vascular bundles, oil glands, and epidermis.

Anatomical structure, flavonoid content, and antioxidant activity of Rhodomyrtus tomentosa leaves and fruits on different age and maturity level

Kuntorini EM, Nugroho LH, Maryani, Nuringtyas TR. 2019. Anatomical structure, flavonoid content, and antioxidant activity of Rhodomyrtus tomentosa leaves and fruits on different age and maturity level. Biodiversitas 20: 3619-3625. Karamunting (Rhodomyrtus tomentosa (Aiton.) Hassk.) is a native plant to southeast Asian countries, and is a well-known medicinal plant used to treat colic diarrhea, wounds, heartburn, abscesses, gynecopathy, and as a pain killer. However, the use of R. tomentosa has not been optimized. This research aimed to observe the anatomical structure, the location, and distribution of flavonoid and to find out the antioxidant activity based on the leaves age and the fruit maturity. Anatomical slides preparation of leaves and fruits were made using the paraffin embedding method with safranin staining. The distribution of flavonoid was analyzed by histochemical test and antioxidant activity was done with DPPH (1,1-diphenyl-2-picrylhydrazyl) method. Leaf anatomical structure shows that the leaf bifacial (dorsiventral) consisted of upper epidermis, mesophyll (palisade and sponge), collateral vascular bundle, parenchyma midrib, abaxial epidermis, globular oil cavity, and non-glandular trichome. Transverse section of green fruit consists of exocarp (thin outer layer), mesocarp (thick inner layer, soft and runny) and endocarp (thin false septa) layers. The histochemical test showed that flavonoid was observed in the leaf epidermis, mesophyll, vascular bundles, secretory cavity, parenchyma and in all fruit types exocarp, mesocarp, endocarp. Antioxidant activity showed that the extract of the young leaves (IC50 = 14.67 ppm) was stronger than the old leaves (IC50 = 19.86 ppm). The antioxidant activity of the purple fruits extract (IC50 = 12.98 ppm) was stronger than the red fruits (IC50 = 28.63 ppm) and the green fruits (IC50 = 48.36 ppm) but it was weaker than quercetin (IC50 = 1.29 ppm). The purple fruit had the highest antioxidant activity compared to other extracts. This information will be useful for developing karamunting as a potential resource of natural antioxidants for functional foods and health products.

Profil Metabolit Skunder Daun Tin (Ficus carica) melalui Analisis Histokimia dan Deteksi Flavonoid dengan Metode Kromatografi Lapis Tipis (KLT)

Tin (Ficus carica) is one of the plants that was mentioned in the Al-Qur’an (at-Tiin 1-3). The plant has the potential of its leaves that can be used as a medicine. In this research were done two tests that are histochemical test and phytochemical test. The method used is descriptive method and the data is processed by descriptive qualitative. Histochemical analysis aims to determine the profile of secondary metabolite compounds in Tin leaf (Ficus carica); among the metabolites were tested the alkaloid, terpenoid, phenol, flavonoid and lipophilic. The second test is the phytochemical test to the examine flavonoid aglycon present, among the tested it is anthocyanin, flavones, flavonol and, biflavonil by thin layer chromatography method. The result showed that the histochemical analysis of Tin leaf (Ficus carica) contain secondary alkaloid, phenol, flavonoid and lipophilic. The phytochemical test result is flavonoid aglycon that show existence is anthocyanin, flavones and biflavonil.Keywords: Tin (Ficus carica), Histochemical profile, Phytochemical test, Flavonoid, TLC.ABSTRAKTin (Ficus carica) adalah salah satu tanaman yang disebutkan di dalam kitab suci Al-Qur’an (Surat At-Tiin 1-3) yang memiliki potensi pada daunnya dapat digunakan sebagai obat. Telah dilakukan penelitian analisis histokimia dan uji fitokimia daun Tin (Ficus carica). Metode yang digunakan dalam penelitian ini ialah metode deskriptif. Data diolah secara deskriptif kualitatif. Analisis histokimia bertujuan untuk mengetahui profil senyawa metabolit sekunder dalam daun Tin (Ficus carica); di antara uji metabolit dilakukan uji alkaloid, terpenoid, fenol, flavonoid dan lipofilik. Uji fitokimia bertujuan untuk memeriksa aglikon flavonoid yang ada, di antara aglikon yang diuji yaitu antosianin, flavon, flavonol dan biflavonil dengan menggunakan metode kromatografi lapis tipis (KLT). Hasil penelitian menunjukkan bahwa analisis histokimia daun Tin (Ficus carica) terdeteksi mengandung senyawa metabolit sekunder alkaloid, fenol, flavonoid dan lipofilik. Hasil uji fitokimia menunjukkan adanya senyawa antosianin, flavon dan biflavonil.Kata kunci: Tin (Ficus carica), Profil histokimia, Uji fitokomia, Flavonoid, KLT.

MORFOLOGI, ANATOMI DAN UJI HISTOKIMIA RIMPANG Curcuma aeruginosa Roxb; Curcuma longa L. DAN Curcuma heyneana Valeton dan Zijp.

Curcuma is a genus of family Zingiberaceae. Its rhizomes, as well as leaves, have long been used in the traditional medicine. This research aimed to examine the morphological and anatomical structure as well as the primary and secondary metabolites of Curcuma aeruginosa Roxb, Curcuma longa L, and Curcuma heyneana Valeton & Zijp. The Anatomical structure, histochemical test and secretory cell density were observed microscopically. The Histochemical test consisted of amilum, protein, lipid, tanin, alkaloid dan flavonoid tests. Observation of anatomical structures of the of rhizome showed that starch grains has a medium size and shape of starch was oval. Rhizomes of Curcuma longa and C. aeruginosa had a positive correlation for starch, protein, lipids, alkaloids, flavonoids and tannins. C. heyneana has the highest density value on protein while C. longa has the highest density value on lipids, alkaloids, flavonoids and tannins.

Histochemistry, content and chemical composition of essential oil in different organs of Alpinia zerumbet

Alpinia zerumbet is a medicinal plant from Asian origin used in folk medicine for the treatment of hypertension, which effect is attributed to terpinen-4-ol, the major component of the essential oil. The objective of this work was to identify the essential oil secretory structures in the leaf, flower, root and rhizome of this plant, and analyze the content and the chemical composition of the oil in the different organs of the plant. Sections were subjected to histochemical test with Nadi reagent for in situ localization of secretion. The essential oil extraction was performed by hydrodistillation in a Clevenger apparatus and the compounds were identified in CG-EM/FID. The histochemical test was positive for terpenoids, confirming the presence of essential oil stored in secretory structures named oils cells present in all analyzed organs. The higher essential oil content was found on the leaf (0.30%), while the petal and the rhizome presented content of 0.10% and 0.06%, respectively. It was not possible to determine essential oil content of the root due to the low amount of biomass produced. There were qualitative and quantitative differences in the chemical composition of the essential oil in the different plant organs, but the major constituent in all of them was the terpinen-4-ol, followed by 1,8 cineol in the leaf and by the α-terpineol in the flower and rhizome.

A histochemical principle bearing on correctness of diagnosis in storage diseases.

A principle is proposed which may help pathologists avoid errors in diagnosis of storage diseases. Tissues from patients in whom a tentative diagnosis of a metabolic disorder has been made often store a number of metabolites in the cells. The presence of these metabolites can occur in single-enzyme or activator defects as a result of the following causes: (a) deposition of metabolites situated near the main substrate of the defective enzyme in the catabolic path, and compounds which were changed after they were deposited; (b) presence of multiple substrates for this enzyme; (c) co-deposition of molecules bound to the main substrates; (d) existence of multiple substrates for a single defective activator molecule. In contrast to these causes, variability in processes not associated with a single-enzyme or activator deficiency may be due to the following: (e) inhibition of multiple hydrolases by drugs or metabolites; (f) localization of substrates and hydrolases in different compartments; (g) multiple enzyme deficiencies; (h) concentration of metabolites beyond the catabolic capacity of cells. According to the proposed principle, diagnosis of storage disease resulting from a single enzyme deficiency can be negated if a wide-range histochemical test shows that the main substrate of a deficient enzyme is not present in some primary storage cells. The validity of the principle and possible pitfalls are discussed.

Sieve tube longevity in white ash (Fraxinus americana) studied with a new histochemical test for the identification of sugar

A histochemical test for functioning sieve tubes, based on the 5-diazouracil sugar reaction, has been used to identify functioning sieve tubes in Fraxinus americana L. The plant material was lyophilized, treated with the reagent, and sectioned in a cryostat. Before the resumption of cambial activity, when buds are still closed (late April – early May), starch grains of the storage tissues in bark and wood disappear and rays, cambrium, and phloem are loaded with sugars. In early spring, sugars begin to accumulate in last year's sieve tubes, especially in those which had not been fully differentiated in the previous active season and those which were functioning but had formed toward the end of the active season. Girdling experiments, carried out during May, showed that translocation in the phloem does occur, to some extent, during the period when the buds develop and the young leaves grow. The reactivated sieve tubes provide the channel of translocation to developing buds for sugars, formed of the dissolution of starch grains. The first early-phloem sieve tubes are formed at the end of May and loaded with sugars during June. There seems to be a short period of simultaneous import into young leaves via reactivated phloem and export via new phloem. After this, last year's sieve tubes are crushed. A noticeable pressure flow does not appear in the main stem until mid-June.