Safety and Risk Assessment of No-Prescription Online Semaglutide Purchases.

This qualitative study assesses the quality, amount of active ingredient, and characteristics associated with counterfeiting of semaglutide purchased from illegal online pharmacies without a prescription.

Same same, but different: exploring the enigmatic role of the pituitary adenylate cyclase-activating polypeptide (PACAP) in invertebrate physiology.

Evidence has been accumulating that elements of the vertebrate pituitary adenylate cyclase-activating polypeptide (PACAP) system are missing in non-chordate genomes, which is at odds with the partial sequence-, immunohistochemical-, and physiological data in the literature. Multilevel experiments were performed on the great pond snail (Lymnaea stagnalis) to explore the role of PACAP in invertebrates. Screening of neuronal transcriptome and genome data did not reveal homologs to the elements of vertebrate PACAP system. Despite this, immunohistochemical investigations with an anti-human PAC1 receptor antibody yielded a positive signal in the neuronal elements in the heart. Although Western blotting of proteins extracted from the nervous system found a relevant band for PACAP-38, immunoprecipitation and mass spectrometric analyses revealed no corresponding peptide fragments. Similarly to the effects reported in vertebrates, PACAP-38 significantly increased cAMP synthesis in the heart and had a positive ionotropic effect on heart preparations. Moreover, it significantly modulated the effects of serotonin and acetylcholine. Homologs to members of Cluster B receptors, which have shared common evolutionary origin with the vertebrate PACAP receptors, PTHRs, and GCGRs, were identified and shown not to be expressed in the heart, which does not support a potential role in the mediation of PACAP-induced effects. Our findings support the notion that the PACAP system emerged after the protostome-deuterostome divergence. Using antibodies against vertebrate proteins is again highlighted to have little/no value in invertebrate studies. The physiological effects of vertebrate PACAP peptides in protostomes, no matter how similar they are to those in vertebrates, should be considered non-specific.

Presence, variation, and potential ecological impact of microplastics in the largest shallow lake of Central Europe.

The presence of microplastics (MPs) in the global ecosystem has generated a rapidly growing concern worldwide. Although their presence in the marine environment has been well-studied, much less data are available on their abundance in freshwaters. MPs alone and in combination with different chemicals has been shown to cause acute and chronic effects on algae and aquatic invertebrate and vertebrate species at different biological levels. However, the combined ecotoxicological effects of MPs with different chemicals on aquatic organisms are still understudied in many species and the reported data are often controversial. In the present study, we investigated, for the first time, the presence of MPs in Lake Balaton, which is the largest shallow lake of Central Europe and an important summer holiday destination. Moreover, we exposed neonates of the well-established ecotoxicological model organism Daphnia magna to different MPs (polystyrene [3 µm] or polyethylene [≤ 100 µm]) alone and in combination with three progestogen compounds (progesterone, drospirenone, levonorgestrel) at an environmentally relevant concentration (10 ng L-1) for 21 days. The presence of 7 polymer types of MPs in the size range of 50-100 µm was detected in Lake Balaton. Similarly to the global trends, polypropylene and polyethylene MPs were the most common types of polymer. The calculated polymer-independent average particle number was 5.5 particles m-3 (size range: 50 µm - 100 µm) which represents the values detected in other European lakes. Our ecotoxicological experiments confirmed that MPs and progestogens can affect D. magna at the behavioral (body size and reproduction) and biochemical (detoxification-related enzyme activity) levels. The joint effects were negligible. The presence of MPs may lead to reduced fitness in the aquatic biota in freshwaters such as Lake Balaton, however, the potential threat of MPs as vectors for progestogens may be limited.

Studies on a widely-recognized snail model species (Lymnaea stagnalis) provide further evidence that vertebrate steroids do not have a hormonal role in the reproduction of mollusks.

Experiments were carried out to determine whether, as with other mollusks that have been studied, the snail, Lymnaea stagnalis, can absorb, esterify and store vertebrate steroids that are present in the water. We also carried out experiments to determine whether neural tissues of the snail could be immunohistochemically stained with an antibody to human aromatase (a key enzyme that catalyzes the conversion of testosterone [T] to 17ß-estradiol [E2]); and, if so, to determine the significance of such staining. Previous studies on other mollusks have reported such staining and have proposed this as decisive evidence that mollusks have the same steroid synthesis pathway as vertebrates. We found that snails absorb, esterify and retain esterified T, E2, progesterone and ethinyl-estradiol (albeit with an absorption rate about four times slower, on a weight basis, than the mussel, Mytilus edulis). We also found that not only anti-human aromatase, but also anti-human nuclear progesterone receptor (nPR) and anti-human gonadotropin-releasing hormone antibodies immunohistochemically stained snail neural cells. However, further experiments, involving gel electrophoretic separation, followed by immunostaining, of proteins extracted from the neural tissue, found at least two positively-stained bands for each antibody, none of which had masses matching the human proteins to which the antibodies had been raised. The anti-aromatase antibody even stained the 140 kDA ladder protein used as a molecular weight marker on the gels. Mass spectrometric analysis of the bands did not find any peptide sequences that corresponded to the human proteins. Our findings confirm that the presence of vertebrate-like sex steroids in molluscan tissues is not necessarily evidence of endogenous origin. The results also show that immunohistochemical studies using antibodies against human proteins are grossly non-specific and likely to have little or no value in studying steroid synthesis or activity in mollusks. Our conclusions are consistent with the fact that genes for aromatase and nPR have not been found in the genome of the snail or of any other mollusk. Our overarching conclusion, from this and our previous studies, is that the endocrinology of mollusks is not the same as that of humans or any other vertebrates and that continuing to carry out physiological and ecotoxicological studies on mollusks on the basis of this false assumption, is an unconscionable waste of resources.

Correlations between Total Antioxidant Capacity, Polyphenol and Fatty Acid Content of Native Grape Seed and Pomace of Four Different Grape Varieties in Hungary.

Grape pomace is a valuable source of various bioactive compounds such as plant-derived polyphenols and polyunsaturated fatty acids (PUFAs). The commercial demand of grape skin and seed powders as nutraceuticals is still growing. However, no distinction is currently made between unfermented native grape seed and grape seed pomace powders regarding their antioxidant activities. Our aim was to find the relationship between the polyphenol and fatty acid content as well as the antioxidant capacity of native and fermented grape seeds of four different grape varieties harvested in the Villány wine region. According to our results, none of the three investigated polyphenols (resveratrol, rutin, quercetin) could be detected in native grape seed samples in correlation with their significantly lower total antioxidant capacities compared to fermented seed samples. Pinot Noir (PN) grape seed pomace samples with the highest resveratrol and oil content showed significantly higher total antioxidant capacity than Cabernet Sauvignon (CS), Syrah (S) and Blue Portugal (BP) samples. Based on the statistical analysis, positive correlation was found between the fatty acid content and the resveratrol concentration in the pomace samples of different grape varieties. In contrast, rutin concentrations were negatively proportional to the fatty acid content of the fermented samples. No significant correlation was found considering the quercetin content of the samples. According to our findings, grape pomace seems a more promising source in the production of nutraceuticals, since it contains polyphenols in higher concentration and exerts significantly higher antioxidant activity than native grape seeds.

Alterations of Nigral Dopamine Levels in Parkinson's Disease after Environmental Enrichment and PACAP Treatment in Aging Rats.

The neuroprotective effects of environmental enrichment and PACAP (pituitary adenylate cyclase-activating polypeptide) are well-described in Parkinson's disease. The aim of our study is to investigate the beneficial effects of these factors in aging parkinsonian rats. Newborn Wistar rats were divided into standard and enriched groups according to their environmental conditions. Standard animals were raised under regular conditions. During the first five postnatal weeks, enriched pups were placed in larger cages with different objects. Aging animals received (1) saline, (2) 6-hydroxidopamine (6-OHDA), or (3) 6-OHDA + PACAP injections into the left substantia nigra (s.n.). On the seventh postoperative day, the left and right s.n. were collected. The s.n. of young and aging unoperated animals were also examined in our experiment. We determined the dopamine (DA) levels by the HPLC-MS technique, while the sandwich ELISA method was used to measure the Parkinson disease protein 7 (PARK7) protein levels. In healthy animals, we found an age-related decrease of DA levels. In aging parkinsonian-enriched rats, the operation did not result in a significant DA loss. PACAP treatment could prevent the DA loss in both the standard and enriched groups. All injured PACAP-treated rats showed remarkably higher protective PARK7 levels. The protective effect of PACAP correlated with the increase of the DA and PARK7 levels.

Lipid and protein tumor markers for head and neck squamous cell carcinoma identified by imaging mass spectrometry.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide. To improve pre- and post-operative diagnosis and prognosis novel molecular markers are desirable. Here we used MALDI imaging mass spectrometry (IMS) and immunohistochemistry (IHC) to seek tumor specific expression of proteins and lipids in HNSCC samples. Among low molecular weight proteins visualized, S100A8 and S100A9 were found to be expressed in the regions of tumor tissue but not in the surrounding healthy stroma of a post-operative microdissected tissue. Marker potential of S100A8 and S100A9 was confirmed by immunohistochemistry of paraffin-embedded pathological samples. Imaging lipids showed a remarkable depletion of lysophosphatidylcholine species LPC[16:0], LPC[18:2] and, in parallel, accumulation of major glycerophospholipid species PE-P[36:4], PC[32:1], PC[34:1] in neoplastic areas. This was confirmed by shotgun lipidomics of dissected healthy and tumor tissue sections. A combination of the negative (LPC[16:0]) and positive (PC[32:1], PC[34:1]) markers was also applicable to uncover tumorous character of a pre-operative biopsy. Furthermore, marker potential of lysophospholipids was supported by elevated expression levels of the lysophospholipid degrading enzyme lysophospholipase A1 (LYPLA1) in the tumor regions of paraffin-embedded HNSCC samples. Finally, experimental evidence of 3D cell spheroid tests showed that LPC[16:0] facilitates HNSCC invasion, implying that HNSCC progression in vivo may be dependent on lysophospholipid supply. Altogether, a series of novel proteins and lipid species were identified by IMS and IHC screening, which may serve as potential molecular markers for tumor diagnosis, prognosis, and may pave the way to better understand HNSCC pathophyisiology.

Mass spectrometric identification of ancient proteins as potential molecular biomarkers for a 2000-year-old osteogenic sarcoma.

Osteosarcoma is the most common primary malignant tumor of bone usually occurring in young adolescent and children. This disease has a poor prognosis, because of the metastases in the period of tumor progression, which are usually developed previous to the clinical diagnosis. In this paper, a 2000-year-old ancient bone remain with osteogenic sarcoma was analyzed searching for tumor biomarkers which are closely related to this disease. After a specific extraction SDS-PAGE gel electrophoresis followed by tryptic digestion was performed. After the digestion the samples were measured using MALDI TOF/TOF MS. Healthy bone samples from same archaeological site were used as control samples. Our results show that in the pathological skeletal remain several well known tumor biomarkers are detected such as annexin A10, BCL-2-like protein, calgizzarin, rho GTPase-activating protein 7, HSP beta-6 protein, transferrin and vimentin compared to the control samples. The identified protein biomarkers can be useful in the discovery of malignant bone lesions such as osteosarcoma in the very early stage of the disease from paleoanthropological remains.

Salivary proteomic analysis of diabetic patients for possible oral squamous cell carcinoma biomarkers.

Since oral squamous cell carcinoma (OSCC) is one of the most important causes of death worldwide, the prevention and early detection plays a crucial role. Recent epidemiological studies have incriminated diabetes as a risk factor for the development of OSCC, as well as oral premalignant lesions. As for the last 20 years diabetes and oral squamous cell carcinoma rates have been increasing rapidly, therefore a reliable detection method of major saliva proteins as possible biomarkers for OSCC is of key priority. In this study we collected whole saliva samples from patients with diabetes and from healthy subjects. To reduce the risk of failure and to keep the investigation good reproducible, we proposed an examination and saliva collecting technique. The proteins were analyzed using SDS-PAGE and MALDI TOF/TOF mass spectrometry. Our findings show that the expression of Annexin A8, Peroxiredoxin-2 and Tyrosine kinase is elevated by patients having diabetes. All these proteins have been previously described in cancer saliva samples also in OSCC. Our current findings showed that testing saliva may be an effective and reliable method for detecting oral cancer in early stages.

Preparation and investigation of LC packing made by microwave-assisted solid-phase synthesis.

The preparation of the so-called bonded phase liquid chromatographic packings is usually carried out by heating the silica, the silane, a catalyst, or a scavenger in an appropriate solvent (often toluene.) Due to the longtime of boiling, the procedure is time and energy consuming, and solvent intensive. The goal of this work is to present a simple, environment-friendly preparation method with reduced solvent consumption to synthetize RP liquid chromatographic stationary phases. The effects of reaction conditions (amount of reagents, composition of the reagent, microwave energy, reaction time, reproducibility of the synthesis) are discussed. Pore structure, surface coverage, the change of the pore structure and surface coverage upon reaction are demonstrated, the efficiency of the column (van Deemter plot for different solutes) is presented. A variety of applications (aromatic hydrocarbons, halobenzenes, bioactive peptides, resveratrol from red wine) demonstrates the separation power of the new phase.

[Removable partial denture design and its effect on remaining teeth, based on Hungarian national survey]. / Részleges lemezes fogpótlások jellemzôi és hatásuk a maradékfogazatra, két szûrôvizsgálat összehasonlított eredményei alapján.

Removable partial dentures (RPD) are used to restore Class 2A, 2B, 2A/1, and Class 3 type partially edentulous arches. Calibrated dentists of the Department of Prosthodontics performed oral health survey first in 1985-1989 and a second survey in 2003-2004. The oral surveys were carried out at the lung cancer screening stations, after obtaining formal consent of those individuals who were originally referred to perform x-ray lung screening examination. Subjects were interviewed and clinically examined under artificial lighting conditions. In the "second" survey data collected on 765 RPDs were analysed. Examined aspects of RPD design were: the protection of the marginal periodontium, extension of major connectors, base plate materials, tooth and tissue support. The level of restoration related damage on the remaining teeth was estimated according to whether remaining teeth were used for retention or for support; had any other type of denture-contact or remaining teeth were free from contact with denture. Approximately 50% of dentures showed dento-mucosal or muco-dental support; metal base was present in 45-48% of dentures; 47% of major connectors were designed with reduced extension; freedom of gingival margin was assured in 43-45% of the dentures. 26.10% of reference teeth; 19.11% of the remaining teeth used for support; 31.53% of the teeth used for retention; 28.07% of the teeth free from contact with denture and 38.00% of those remaining teeth with any other type of denture contact showed a sign of damage. Findings of the authors' survey show a considerable improvement in all aspects of RPD design--but still further progression is required. Those principles of RPD design taught in dental schools can only gradually influence the routine daily dental practice. Assessment of remaining teeth damages confirm the standards we outlined for RPD design with the main priorities of dental support and the protection of the gingival margin.

[The prevalence of oral mucosal lesions in Hungary]. / Szájnyálkahártya-elváltozások elofordulási gyakorisága Magyarországon.

The aim of the survey was to recognize the lesions of the oral mucosa (any alterations from healthy normal tissues) related to gender, age, intraoral localization, geographic regions, according to smoking habits and dental surgery attendance, based on the data of "Epidemiological national survey on oral health 2003-2004.". In the nationwide survey 4606 people (2923 women, 1683 men) participated. Oral mucosal lesions registered in 10.14 and 9.41% in males and females (respectively). The highest prevalence of mucosal lesions occurred in 45-64 year old age group. The highest number of the oral lesions was found in the buccal mucosa in males (42%). In females the most of the lesions are registered on the hard palate (24%) and on the buccal mucosa (24%). By geographic regions the highest prevalence of oral mucosal lesions occurred in "Közép-Dunántúl" statistical region, and the lowest prevalence was found in "Dél-Dunántúl". Statistically significant correlation could be shown between prevalence of oral mucosal lesions and smoking habits (p < or = 0.05) and there was a significant correlation between the frequency of dental surgery attendance and the prevalence of mucosal lesions in the examined population. The data of this epidemiological study could serve a good base for planning of preventive programs in different populations in the future.

[Neurologic complications of aortic dissection]. / Az aortadissectio neurológiai szövódményei.

INTRODUCTION: Beside the damages of the cardiovascular system the lesions of the the nervous system are the most common complications of aortic dissection. This is usually an early event, therefore the dissection of the aorta may manifest itself as an acute primary neurologic disease. The aim of this study is to describe the frequency and distribution of acute neurologic symptoms occurring in aortic dissection and the distribution of their clinico-pathologic features and to establish correlations between these and the acute in-hospital mortality as well as to discuss available diagnostic and therapeutic possibilities. PATIENTS AND METHODS: The study was based of 95 cases of acute dissection of aorta (with additional three later events of redissection), observed in a longitudinal study over a period of 29.5 years, in a population of 106,000 (in Western Hungary). RESULTS: Of the 95 patients 20 (21%) died before admission. Neurological complications were observed in 30 of the 75 patients admitted to hospital (40%). Symptoms involving the central nervous system were found in 24 patients, affecting the spinal cord in two and the peripheral nervous system in four cases. The dissection of the aorta was diagnosed in vivo only in 22 out of the 75 patients who died in hospital (29%). 53 patients (71%) without correct diagnosis received supportive therapy only. The average survival time of the 21 patients with proximal dissection of aorta was 48.5 hours. The survival time of 23 patients with the same type of dissection involving the vessels of the aortic arch was 22.2 hours. This difference in survival time was significant (p = 0.0152). 20 of 23 patients (87%) in this group showed signs of neurologic damage confirming earlier experience that neurological complications can seriously worsen the otherwise already catastrophic prognosis of aortic dissection. CONCLUSIONS: The study brought compelling evidence for the need for early diagnosis and rapid transfer of patients to appropriate cardiac surgery centers for definitive diagnosis and therapy.