Trichrome Staining Techniques Research Articles

Detection of Parasites in Locally Sourced Fresh Fruits and Vegetables Using Various Diagnostic Techniques

A prospective study was conducted from September 2022 to March 2023 to examine the occurrence and contamination rates of different parasitic stages on fresh edible fruits and vegetables in local markets of Erbil city. Eight different types of fruits and vegetables were selected from eight markets, and 144 samples were analyzed for parasites using direct sedimentation, indirect floatation (ZnSO4), Kinyoun, and trichrome staining methods. The highest total frequencies of parasites were detected using the direct sedimentation method (18.75%), followed by the indirect floatation method (6.25%), Kinyoun technique (5.55%), and modified trichrome staining (4.16%). A. lumbricoides (Ova) was detected with 4.16% using direct sedimentation, while no positive detections were observed with other techniques. Kinyoun technique identified Cryptosporidium spp. oocysts. For E. histolytica 1.3% were positive using the direct sedimentation method, while the other three methods showed no positive detections. Similar results were observed for G. lamblia where only the modified trichrome staining technique detected 0.69% with trophozoites and 1.3% with cysts. Additionally, the direct sedimentation method revealed the presence of nematodes at 2.77%, but no positive detections were made with other methods. Concerning Taeniid eggs, the indirect floatation identified 0.69% with ova, while other methods showed no positive detections. Lastly, the direct sedimentation method yielded 6.25% with unidentified flagellate protozoa, but no positive detections were found using the indirect floatation method, Kinyoun technique, or modified trichrome staining technique. The study concluded that Kinyoun and trichrome techniques are the gold standard for detecting different parasite stages.

Prevalence and Risk Factors of Giardia intestinalis Infestation and Assemblage of Isolates Among Monastery Primary School Children in Yangon, Myanmar.

Giardiasisis one of the major causes of diarrhea among children. To determine the prevalence, risk factors, and genotype of Giardia intestinalis, a cross-sectional descriptive study was done on stool samples of 462 children attending three monastery primary schools from North Okkalapa Township in Yangon, Myanmar from January 2016 to February 2019. Socioeconomic data were collected using a pre-tested questionnaire after obtaining informed consent. Direct wet mount, formalin-ether sedimentation, and trichrome staining techniques were used for the primary identification and then molecular identification was carried out by conventional polymerase chain reaction(PCR)-sequencing assay. G. intestinalis was identified in 11.7% (54/462) of students. There was no significant association with water source (p=0.948) and drinking untreated water (p=0.595). The infection was more common in children with low-educated parents, unsanitary garbage disposal practices, and no restrooms. All isolates were G. intestinalis assemblage B. This is the first study characterizing human isolates in a lower region of Myanmar, at the molecular level [MOU1]. These findings pointed out the high prevalence of G. intestinalis among primary school children from densely populated and low-resource settings.

The fulcrum of the human heart (Cardiac fulcrum)

Introduction: The functional anatomy of the myocardium allows envisioning that it initiates and ends at the origin of the great vessels. In our research, we have always considered that it should have a point of attachment to allow its helical rotation to fulfill the fundamental movements of shortening-torsion (systole) and lengthening-detorsion (suction), which once found, was called the cardiac fulcrum. Material and Methods: A total of 31 hearts, arising from the morgue and slaughterhouse were used: 17 corresponded to bovids and 14 were human. Anatomical and histological studies were performed. The heart was fixed in 10% buffered formalin. Hematoxylin-eosin, Masson’s trichrome staining technique and 4-micron sections were used for the histological study, and 10 % formalin was used as buffer. Results: The anatomical investigations have revealed that all the hearts (bovids and humans) have a myocardial support whose histological structure in the analyzed specimens presents with an osseous or chondroid-tendinous character. In this structure, that we have call cardiac fulcrum, are inserted the myocardial fibers at the origin and end of the band, which correspond to the continuous myocardium coiled as a helix. Conclusion: This description of the fulcrum would end the problem of lack of support of the myocardium to fulfill its function of suction/ejection.

The Fulcrum of the Human Heart (Cardiac fulcrum)

Introduction: The functional anatomy of the myocardium allows envisioning that it initiates and ends at the origin of the great vessels. In our research, we have always considered that it should have a point of attachment to allow its helical rotation to fulfill the fundamental movements of shortening-torsion (systole) and lengthening-detorsion (suction), which once found, was called the cardiac fulcrum. Materials and methods: A total of 31 hearts, arising from the morgue and slaughterhouse were used: 17 corresponded to bovids and 14 were human. Anatomical and histological studies were performed. The heart was fixed in 10% buffered formalin. Hematoxylin-eosin, Masson’s trichrome staining technique, and 4-micron sections were used for the histological study, and 10 % formalin was used as a buffer. Results: The anatomical investigations have revealed that all the hearts (bovids and humans) have myocardial support whose histological structure in the analyzed specimens presents with an osseous or chondroid-tendinous character. In this structure, which we have called the cardiac fulcrum, are inserted the myocardial fibers at the origin and end of the band, which correspond to the continuous myocardium coiled as a helix. Conclusion: This description of the fulcrum would end the problem of lack of support of the myocardium to fulfill its function of suction/ejection.

Polycaprolactone/Testicular Extracellular Matrix/Graphene Oxide-Based Electrospun Tubular Scaffolds for Reproductive Medicine: Biomimetic Architecture of Seminiferous Tubules.

Numerous scaffolds are developed in the field of testicular bioengineering. However, effectively replicating the spatial characteristics of native tissue, poses a challenge in maintaining the requisite cellular arrangement essential for spermatogenesis. In order to mimic the structural properties of seminiferous tubules, the objective is to fabricate a biocompatible tubular scaffold. Following the decellularization process of the testicular tissue, validation of cellular remnants' elimination from the specimens is conducted using 4',6-diamidino-2-phenylindole staining, hematoxylin and eosin staining, and DNA content analysis. The presence of extracellular matrix (ECM) components is confirmed through Alcian blue, Orcein, and Masson's trichrome staining techniques. The electrospinning technique is employed to synthesize the scaffolds using polycaprolactone (PCL), extracted ECM, and varying concentrations of graphene oxide (GO) (0.5%, 1%, and 2%). Subsequently, comprehensive evaluations are performed to assess the properties of the synthetic scaffolds. These evaluations encompass Fourier-transform infrared spectroscopy, scanning electron microscopy imaging, scaffold degradation testing, mechanical behavior analysis, methylthiazolyldiphenyl-tetrazolium bromide assay, and in vivo biocompatibility assessment. The PCL/decellularized extracellular matrix with 0.5% GO formulation exhibits superior fiber morphology and enhanced mechanical properties, and outperforms other groups in terms of in vitro biocompatibility. Consequently, these scaffolds present a viable option for implementation in "in vitro spermatogenesis" procedures, holding promise for future sperm production from spermatogonial cells.

Frequency of Entamoeba Complex in Individuals Referred to the Medical Laboratories in Jahrom City, South of Iran.

This study aimed to determine the prevalence of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii, using microscopic and molecular methods in Jahrom city (Fars Province), south of Iran. Stool samples were collected from 360 outpatients referred to the medical laboratories in Jahrom city. Standard parasitological methods, including direct wet mount examination, formalin-ether sedimentation technique, and trichrome staining techniques, were used for detection of Entamoeba complex. Nested polymerase chain reaction (PCR) was used to identify the genus/species of Entamoeba. Entamoeba complex cysts were detected in 2.5% (9/360) of samples by microscopic methods, while 11 isolates were considered positive for the Entamoeba complex by nested PCR. Among them, 2 (18.18%) out of 11 samples were E. histolytica, 5 (45.45%) were E. dispar, and 4 (36.36%) were E. moshkovskii. Molecular positivity was more prevalent among females (4.16%), people living in rural areas (3.44%), and people over 60 years old (13.33%). Considering the clinical manifestations, the Entamoeba complex infection in patients with fever (7.69%), severe diarrhea (5.10%), and nausea (5%) was high. This study reported the presence of E. histolytica, E. dispar, and E. moshkovskii in Jahrom city. Therefore, we suggest more public health interventions in Jahrom city.

Prevalence and risk factors for infection with Entamoeba histolytica/dispar/ moshkovskii complex in people living in the slightest and outermost islands of Indonesia.

Entamoeba histolytica (E. histolytica), the causative agent of amoebiasis, is still a global public health problem that cannot be controlled, especially in tropical and subtropical countries. This study was conducted to obtain information about the incidence of Entamoeba histolytica/dispar/ moshkovskii complex infection and the factors that influence it. The prevalence of infection with the Entamoeba histolytica/dispar/moshkovskii complex and the factors that influence it in people living on the smallest and outermost island of Indonesia, Sabang Island, Aceh Province. This study involved 335 respondents aged >= 10 years. Respondents were selected by non-probability sampling technique. Interviews and observations were conducted to identify risk factors. The Entamoeba histolytica/dispar/ moshkovskii complex was identified by direct examination, concentration, and Whitley's trichrome staining techniques. A Chi-Square test was performed to analyze the correlation of risk factors with the incidence of infection. The prevalence of infection with the Entamoeba histolytica/dispar/ moshkovskii complex in the people of Sabang Island was 26.6% (89/335). Source and adequacy of clean water correlated with the incidence of Entamoeba histolytica/dispar/moshkovskii complex infection. Demographic variables are not correlated with the incidence of infection. However, the group of women aged > 61 years, unemployed, unmarried, and earning less than the regional minimum wage tend to be more likely to be found with Entamoeba histolytica/dispar/moshkovskii complex infections. Thus it can be concluded that the prevalence of infection with the Entamoeba histolytica/dispar/moshkovskii complex on Sabang Island is in the high category. The prevalence of E. histolytica as the causative agent of amoebiasis cannot be explained with certainty because the two identical non-pathogenic Entamoeba species cannot be distinguished by microscopic identification. Sources and adequacy of clean water correlate with the incidence of Entamoeba histolytica/dispar/moshkovskii complex infection in the people of Sabang Island.

Investigating intestinal parasitic infections with emphasis on molecular identification of Strongyloides stercoralis and Trichostrongylus colubriformis in north of Iran

Currently, parasitic infections are one of the important health problems in the world, especially in developing countries. This study aims to investigate intestinal parasites with an emphasis on molecular identification through the analysis of mitochondrial COX1 and ITS2 gene sequences of Strongyloides stercoralis (S. stercoralis) and Trichostrongylus spp. in north of Iran. Five hundred forty stool samples were collected from medical diagnostic laboratories affiliated with Mazandaran University of Medical Sciences in Sari city, north of Iran. First, all the samples were examined using direct smear, formalin-ether sedimentation, and trichrome staining technique. Suspected samples of Strongyloides larvae were cultured in agar plate. Then, DNA was extracted from samples containing Trichostrongylus spp. eggs and Strongyloides larvae. To amplify DNA, PCR was performed and the samples with a sharp band in electrophoresis were sequenced by Sanger method. Overall, the prevalence of parasitic infections in the study population was 5.4%. The highest and the lowest level of infection was observed with Trichostrongylus spp. and S. stercoralis at 3% and 0.2%, respectively. No traces of live Strongyloides larvae were seen in the culture medium of the agar plate. The six isolates obtained from the amplification of the ITS2 gene of Trichostrongylus spp. were sequenced, all of which were Trichostrongylus colubriformis. The sequencing results of COX1 gene indicated S. stercoralis. In the present study, the prevalence of intestinal parasitic infections in north of Iran has relatively decreased that its main reason can be due to the coronavirus epidemic and compliance with health principles. However, the prevalence of Trichostrongylus parasite was relatively high that it requires special attention to apply appropriate control and treatment strategies in this field.

The effect of green robust coffee gel (coffea canephora) to collagen density on gingival wound healing (an in-vivo study in wistar rats)

Objective: This study aims to determine the effect of robusta green coffee gel on the collagen density in Wistar rats. Material and Methods: The subjects were 45 Wistar rats, each wounded with a 3 mm punch biopsy in the mandibular incisor gingiva. The rats were then divided into 5 groups, namely the positive control group (Povidone-iodine 10%), negative control (gel base), and 3 treatment groups (10%, 20%, and 40% concentration of Robusta green coffee gel). The intervention was carried out twice a day. A total of 3 mice in each group were then sacrificed on the 5th and 10th days to see the collagen density using the Mallory Trichrome staining technique. Collagen density was observed using a microscope with 400x magnification in 5 fields of view. Kruskal Wallis test was used for data processing. Results: There was no difference in collagen density regardless of the coffee gel concentration at each observation time, with p>0.05. Conclusion: There was no effect on the times of observation or dose concentration of robusta coffee gel on collagen density on gingival wound healing in Wistar rats.

The vascular protective effects of trihoney in hypercholesterolemic atherosclerotic rabbits, a comparative study with atorvastatin

ABSTRACT Atherosclerosis is a chronic inflammatory disease of the inner wall of large- and medium-sized arteries. The histopathological lesion of atherosclerosis is the atherosclerotic plaque. This study aimed to investigate the vascular protective effects of trihoney against the development and progression of atherosclerotic plaques. Thirty male New Zealand white (NZW) rabbits were grouped into six groups, comprising five rabbits per group as follows: normal diet (C), normal diet with 0.6 g/kg/day of trihoney (C + H), 1% cholesterol diet (HCD), 1% cholesterol diet with 0.3 g/kg/day of trihoney (HCD+H1), 1% cholesterol diet with 0.6 g/kg/day of trihoney (HCD+H2) and 1% cholesterol diet with 2 mg/kg/day of atorvastatin (HCD+At). Animals were sacrificed after 12 weeks of treatment. Atherosclerotic plaques were quantified, measured and examined using Sudan IV, Hematoxylin and Eosin (H&E), Verhoeff-van Gieson (VGS) and Masson’s trichrome staining techniques. Trihoney resulted in a significant (p < 0.05) reduction in the number of atherosclerotic lesions, tunica intima thickness, and tunica intima/tunica media thickness ratio. Trihoney preserved the orientation of medial elastic tissues and exhibited a plaque stability effect in the form of subendothelial smooth muscle cells (SMCs) recruitment and reduction of collagen deposition in tunica media. Trihoney could be recommended as a protective natural product and a future adjuvant to statins for the management of atherosclerotic plaques.

Status of Giardia Infection with Other Intestinal Parasites among Paediatric Patients in Tertiary Medical College Hospitals

Background: Paediatric diarrhoea, the burning issue of nowadays, drastically affects the child mortality and morbidity. Apart from virus and bacteria, parasites also play an important role in causing diarrhoea at that age group. Giardiasis, the most prevalent among the parasitic infections, though neglected yet carries significance because of its long term health issues. The aim of the study was to identify the intestinal parasites by wet mount preparation and further detection of the flagellated protozoan parasite, Giardia by special staining - Trichrome staining technique.&#x0D; Materials and methods: It was an observational cross sectional study which was carried between the periods of July 2019 to June 2020, where a total of 200 diarrhoeic patients up to 18 years were enrolled. Direct microscopic method was done where parasites were detected by wet mount preparation and further Trichrome stainning was performed for the detection of Giardia. &#x0D; Results: Out of 200 faecal samples, cysts of Giardia, ova of Ascaris Lumbricoides (AL) Trichuris Trichiura (TT) and both AL and TT were identified in 2.5%, 1.5%, 0.5% and 1.0% samples respectively by wet mount preparation. By Trichrome staining method we detected 3.5% of Giardia cysts.&#x0D; Conclusion: Staining technique could be applied in addition to wet mount preparation of stool samples. So that we could recover the undetected Giardia cyst in the aforesaid samples.&#x0D; JCMCTA 2021 ; 32 (2) : 111-114

SAFETY PROFILE OF MUSA PARADISIACA AQUEOUS FRUIT- EXTRACT IN TESTIS OF NORMAL RAT

Musa paradisiaca (plantain) is one of the common staple foods in East Africa. The leaves, roots, and fruits have been a revolutionary traditional remedy in the management of male sexual infertility. This study aimed at evaluating the safety profile of plantain aqueous fruit-extract in the testis of male rats. Thirty Adult Wistar albino rats weighing about 100–150 g in body weight (bw) were used in this study. They were divided into three (3) groups of 10 animals each (n=10). Groups I received water only. Groups II and III received low (200 mg/kg bw) and high (400 mg/kg bw) doses of plantain, respectively. The animals were allowed free access to rat feed and water. The extract was given once daily for 21 days via the oral route. The histomorphology of the testis was performed using Masson’s Trichrome staining technique, while the sperm profile was analyzed using a Makler counting chamber with Olympus Microscope. Plantain administration in normal rats significantly increased the body weights of the rats. Seminal analysis showed improved sperm parameters in treated rats, but rats in the highdose group presented a high level of agglutination compared to rats in the control and low -dose groups. Histological observations showed mild-to-moderate proliferation of cells in the testicular germ layer of the treated rats, indicating enhanced spermatogenesis. Testicular morphology in the group that received high dose showed mild degenerative changes in their testicular germ layer. Plantain, at moderate consumption can improve testicular activity and reproductive health in male while at a high consumption rate it may cause mild degenerative changes in the testicular germ layer.

Ceftriaxone and Melittin Synergistically Promote Wound Healing in Diabetic Rats.

High glucose levels in diabetic patients are implicated in delay wound healing that could lead to more serious clinical complications. The aim of the present work was to examine the formulation of ceftriaxone (CTX) and melittin (MEL) as nanoconjugate (nanocomplex)-loaded hydroxypropyl methylcellulose (HPMC) (1.5% w/v)-based hydrogel for healing of acute wounds in diabetic rats. The CTX–MEL nanoconjugate, formulated by ion-pairing at different molar ratio, was characterized for size and zeta potential and investigated by transmission electron microscopy. CTX–MEL nanoconjugate was prepared, and its preclinical efficacy evaluated in an in vivo model of acute wound. In particular, the potential ability of the innovative CTX–MEL formulation to modulate wound closure, oxidative status, inflammatory markers, and hydroxyproline was evaluated by ELISA, while the histopathological examination was obtained by using hematoxylin and eosin or Masson’s trichrome staining techniques. Quantitative real-time PCR (qRT-PCR) of the excised tissue to measure collagen, type I, alpha 1 (Col1A1) expression and immunohistochemical assessment of vascular endothelial growth factor A (VEGF-A) and transforming growth factor beta 1 (TGF-β1) were also carried out to shed some light on the mechanism of wound healing. Our results show that the CTX–MEL nanocomplex has enhanced ability to regenerate epithelium, also giving better keratinization, epidermal proliferation, and granulation tissue formation, compared to MEL, CTX, or positive control. The nanocomplex also significantly ameliorated the antioxidant status by decreasing malondialdehyde (MDA) and increasing superoxide dismutase (SOD) levels. The treatment of wounded skin with the CTX–MEL nanocomplex also showed a significant reduction in interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) pro-inflammatory cytokines combined with a substantial increase in hydroxyproline, VEFG-A, and TGF-β1 protein expression compared to individual components or negative control group. Additionally, the CTX–MEL nanocomplex showed a significant increase in mRNA expression levels of Col1A1 as compared to individual compounds. In conclusion, the ion-pairing nanocomplex of CTX–MEL represents a promising carrier that can be topically applied to improve wound healing.

Comparison of diagnostic methods (wet mount, trichrome staining, formol-ether, PCR, and xenic in vitro culture) for the detection of Blastocystis in stool samples in Urmia educational hospitals, the Northwest of Iran.

Blastocystis spp. is known as a common intestinal protozoan parasite in human and animals. The parasite has a worldwide distribution and is frequently detected in faecal samples in clinical parasitology laboratories. The goal of the study was to compare the sensitivity and specificity of formol-ether technique (FECT), trichrome staining, xenic in vitro culture (XIVC), microscopy of faecal smears, and polymerase chain reaction (PCR) methods for detecting Blastocystis spp. in human stool samples. The prevalence of the parasite in the stool samples referred to educational hospitals was also determined. A total of 575 cases were assessed to detect the parasite. After collecting from patients referring to Urmia educational hospitals, the samples were examined by microscopy of faecal smears, trichrome staining, FECT, XIVC using Jones' medium, and PCR, to evaluate the presence of Blastocystis spp. Microscopy of faecal smears, trichrome staining, FECT, and PCR technique detected 94, 100, 96, and 44 positive cases, with the sensitivity of 71.3%, 74.4%, 74.4%, and 80.4% and the specificity of 99.6%, 99.1%, 100%, and 93.1%, respectively. XIVC method identified the highest number of positive cases (129 cases) among the other methods. Our findings indicates that XIVC technique is more sensitive method for the detection of Blastocystis spp. in human stool, as compared to direct smear, trichrome staining, FECT, and PCR methods.

In Vitro and In Vivo Osteogenesis Induced by Icariin and Bone Morphogenetic Protein-2: A Dynamic Observation.

In the present study, we aimed to compare the effects of icariin (ICA) and bone morphogenetic protein-2 (BMP-2) on osteoblast proliferation and osteogenesis in bone defects. We found that in vitro ICA or BMP-2 treatment is able to increase osteoblast proliferation, which was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Specifically, ICA at a concentration of 30 µg/ml had the strongest ability to promote cell proliferation, which is equivalent with the effect of BMP-2 at a concentration of 50 µg/ml. Furthermore, Western blot and RT-qPCR analyses showed that treatment with ICA (20–30 µg/ml) had similar increase effect with BMP-2 (50 µg/ml) on the protein and mRNA levels of BMP-2, osteoprotegerin (OPG), and alkaline phosphatase (ALP) mRNAs. In addition, the animal model of bone defects was successfully prepared. The in vivo data showed that compared with the control group, highest osteogenesis in the ICA or BMP-2 groups was observed at different observational times. Four weeks after surgery, osteogenesis in the BMP-2 group was slightly higher than that in the ICA group, but there was no significant difference between the two groups until the eighth week. ICA promotes osteoblast proliferation by stimulating the expression of BMP-2 and OPG proteins and upregulating the expression of BMP-2, OPG, and ALP mRNAs. ICA at a certain concentration has the same osteogenic effect as BMP-2. ICA or BMP-2 composite nanomaterials can be used as a framework to guide bone regeneration and promote osteogenesis. In addition, the combined use of hematoxylin-eosin and Goldner’s trichrome staining techniques contributes to acquiring better bone morphometric information about bone defects.

New trichrome stains identify cysts of Colpodella sp. (Apicomplexa) and Bodo caudatus.

Colpodella species are free-living close relatives of apicomplexans that were recently reported to cause red blood cell infection in an immunocompromised human host and in a tick-borne human infection resulting in neurological symptoms. Unambiguous identification of the life cycle stages of Colpodella sp. using routine stains for light microscopy will aid rapid diagnosis in infections. Similarly, cells in culture and environmental samples can be rapidly identified by staining. Staining protocols are currently unavailable for cell detection by light microscopy. In this study, we investigated the feasibility of performing routine staining techniques for light microscopy for differentiating Colpodella sp. (ATCC 50594) and Bodo caudatus cysts in Hay medium cultures. We tested different basic and acidic dyes alone and in combination and also utilized a commercial trichrome staining protocol. The nonspecific fluorescent dye Calcofluor white was also evaluated. Staining times, dye concentrations, use of tap or distilled water rinses, use of a mordant and inclusion, or omission of decolorizers after staining were evaluated. We compared the intensity of color, clarity of morphological features, and cytoplasmic structures detected after staining. We report a new trichrome staining technique that allowed clear identification and differentiation of cyst stages of Colpodella sp. and B. caudatus. Immature Colpodella sp. cysts were identified as having an irregular, dual-colored (demilune), dark blue-purple and white appearance. Mature Colpodella sp. cysts stained dark red-blue and were identified in four-way mitotic division, while cysts of B. caudatus in diprotist or monoprotist (ATCC 30905) cultures were detected as spherical and red-pink in appearance.

Glucose Fluctuations Promote Aortic Fibrosis through the ROS/p38 MAPK/Runx2 Signaling Pathway

Aim: Glucose fluctuations may be responsible for, or further the onset of arterial hypertension, but the exact mechanisms remain unclear. The purpose of this study was to investigate the mechanisms behind and related to aortic fibrosis and aortic stiffening induced by glucose fluctuations. Methods: Sprague-Dawley rats were injected with streptozotocin (STZ) and randomly divided into three treatment groups: controlled STZ-induced diabetes (C-STZ); uncontrolled STZ-induced diabetes (U-STZ); and STZ-induced diabetes with glucose fluctuations (STZ-GF). After 3 weeks, rat blood pressure (BP) was tested, and aortic fibrosis was detected by using the Masson trichrome staining technique. Levels of p38 mitogen-activated protein kinase (p38 MAPK), runt-related transcription factor 2 (Runx2), collagen type 1 (collagen I), and NADPH oxidases were determined by Western blot.Rat vascular smooth muscle cells in vitro were used to explore underlying mechanisms. Results: The systolic BP of diabetic rats in the C-STZ, U-STZ, and STZ-GF groups was 127.67 ± 6.53, 150.03 ± 5.24, and 171.63 ± 3.53 mm Hg, respectively (p< 0.05). The mean BP of diabetic rats in the three groups was 91.20 ± 10.07, 117.29 ± 4.28, and 140.58 ± 2.14 mm Hg, respectively (p< 0.05). The diastolic BP of diabetic rats in the three groups was 73.20 ± 12.63, 101.93 ± 5.79, and 125.37 ± 4.62 mm Hg, respectively (p< 0.05). The ratios of fibrosis areas in the aortas of the three groups were 11.85 ± 1.23, 29.00 ± 0.87, and 48.36 ± 0.55, respectively (p< 0.05). The expressions of p38 MAPK, Runx2, and collagen I were significantly increased in the STZ-GF group. In vitro, applications of inhibitors of reactive oxygen species (ROS) and p38 MAPK successfully reversed glucose fluctuations that would have possibly induced aortic fibrosis. Conclusions: Blood glucose fluctuations aggravate aortic fibrosis via affecting the ROS/p38 MAPK /Runx2 signaling pathway.

Ethanol as Dentin Pretreatment on the Bonding Performance of a Two-Step Etch-and-Rinse Adhesive: An In Vivo Study.

Objective The aim of this study was to evaluate the use of 70 vol.% ethanol as a dentin pretreatment on the bond strength (BS) of a two-step etch-and-rinse adhesive. Materials and Methods Resin composite Class I restorations were clinically bonded to acid-etched dentin of human sound third molars using Adper Single Bond 2 (SB, 3M ESPE) and randomly divided into two major groups: dentin saturated with water (control) or 70 vol.% ethanol (ethanol). The teeth were divided into two subgroups: immediately extracted and tested after 24 hours and extraction after 18 months. Bonded teeth (Adper SB 2) were cut into resin–dentin sticks that were tested by microtensile BS, and the failure mode was thereafter evaluated. Statistical Analysis Data were statistically analyzed using two-way analysis of variance and Holm–Sidak post hoc test (α = 0.05). Additional bonded resin–dentin slabs from each group were examined under light microscopy (LM) using the Masson's trichrome staining technique. Results The lowest BS was obtained by ethanol pretreated dentin after aging, while other groups presented similar BS. The LM analysis showed the presence of resin-sparse collagen fibrils in groups examined immediately (24 hours) and the presence of several gaps due to collagen degradation at the interfaces of ethanol pretreated aged specimens (18 months). Conclusion The use of 70 vol.% ethanol in dentin before the application of a two-step etch-and-rinse adhesive should be avoided once it resulted in a significant drop on the BS.

Frequency, associated factors and clinical symptoms of intestinal parasites among tuberculosis and non-tuberculosis groups in Iran: a comparative cross-sectional study.

Co-infection of human tuberculosis (TB) and intestinal parasites infections (IPIs) is a public health problem, especially in low- and middle-income countries. There is no data on this issue in Iran. Therefore, we investigated the prevalence of IPIs among patients with TB in Iran. Stool samples were collected from 161 patients with TB and 181 healthy people (non-TB group). Standard parasitological methods including direct slide smear, formalin-ether concentration, trichrome, modified Ziehl-Neelsen and chromotrope 2R staining techniques were used for detection of intestinal protozoa and helminths. Nested-PCR and sequence analysis were used to identify the genotypes of Cryptosporidium and human-infecting species of microsporidia. Data analysis was performed using SPSS version 16. The frequency of IPIs in the non-TB group (16.5%) was slightly lower than in patients with TB (21.1%), although statistical significance was not observed (OR, 0.74; 95% CI, 0.43-1.27; P= 0.28). Blastocystis (11.8%) was the most common parasite detected in patients with TB. Infection with multiple parasites in the non-TB group (2.2%) was significantly lower than in patients with TB (7.5%) (OR, 0.28; 95% CI, 0.08-0.88; P=0.02). The ova of Taenia spp., Ascaris lumbricoides and Hyamenolepis nana were identified in three patients with TB (1.9%), while only one person (0.5%) in the non-TB group was infected with Enterobius vermicularis. The results of genotyping revealed two C. parvum subtype families (IIa and IId) and three E. bieneusi genotypes (Ebcar4, IH and jLD-1). Our results showed a higher prevalence of IPIs in patients with TB in comparison with non-TB subjects. Moreover, our findings suggest a proper health education program for good personal hygiene habits, and also preventative measures to avoid the acquisition of IPIs in patients with TB.

Farnesyl Pyrophosphate Synthase Blocker Ibandronate Reduces Thoracic Aortic Fibrosis in Diabetic Rats

BackgroundThis study assessed the effect of ibandronate (IBN), a farnesyl pyrophosphate synthase (FPPS) inhibitor, on vascular remodeling in diabetic rats. MethodsA rat model of diabetes was induced by a high-fat and high-sugar diet combined with a small dose of streptozotocin. The diabetic rats received 5 µg/kg of ibandronate solution or normal saline subcutaneously every morning for 16 weeks. The morphology of the thoracic aorta was assessed by hematoxylin and eosin and Masson's trichrome staining techniques. Gene expression levels of connective tissue growth factor (CTGF) and FPPS were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. CTGF and FPPS protein levels were determined by Western blotting analysis. ResultsRats with diabetes mellitus showed moderate hyperglycemia, insulin resistance, hyperlipidemia and thoracic aortic fibrosis. FPPS was significantly upregulated in the thoracic aorta from diabetic animals. Interestingly, IBN treatment for 16 weeks alleviated the diabetes-induced histopathologic changes in the thoracic aortic wall and reduced CTGF protein and mRNA levels. ConclusionsThese findings provided evidence that FPPS is involved in thoracic aortic fibrosis in diabetic rats. Meanwhile, IBN could alleviate vascular remodeling in diabetic animals.