Annosvaihtelu ex tempore -sotalolihydrokloridikapseleissa sekä ravintoletkun läpi johdetuissa kapselisisällöissä

Nowadays, there is still lack of commercially produced drugs for children. Extemporaneous compounding is needed widespread. Oral powders, capsules and oral suspensions are the most typical extemporaneous dosage forms. In Finland, oral powders have traditionally been the most used. Major concern relating to the extemporaneous products is that they are not authorized. That means that their safety and effectiveness have not been established. Compounding oral powders is time consuming and their overall mass is much higher compared to capsules with same strength. That increases the amount of foreign matter in child patients, which is highly not recommended. The aim of this study was to examine, whether the extemporaneous sotalolhydrochloride capsules meet European Pharmacopoeia standards of content uniformity. Additionally, because the feeding tubes are widely used in neonatal patients, it was also reasoned to examine the content uniformity of capsules lead through the feeding tube. A significant part of this study was to develop an accurate and effective HPLC -method for analyzing sotalolhydrochloride, which, in the end, turned out well. With its seven minute driving time per sample, it is suitable even in routine analysis. Two of three capsule batches, as well as the oral powders, met the European Pharmacopoeia standards of content uniformity. Also, leading the capsule contents through the feeding tubes met the standards, but the amount of drug substance was significantly lower compared to capsules and oral powders. With lower overall mass and being quicker to prepare, capsules are recommendable option for traditional oral powders in extemporaneous children's medication. Still, according to this study, it is important to take into consideration the possibility of excessive variation in content uniformity. Thus, in the future, it is necessary to develop the quality control systems in hospital pharmacies.Lapsille tarkoitettuja lääkevalmisteita on markkinoilla edelleen suhteellisen vähän. Tämän vuoksi lastenlääkinnässä joudutaan usein turvautumaan ex tempore -lääkevalmistukseen. Yleisimmin tämä tarkoittaa annosjauheiden, oraalisuspensioiden tai kapseleiden valmistamista aikuisille tarkoitetuista kaupallisista valmisteista. Suomessa on perinteisesti valmistettu annosjauheita. Ongelmallista kaikissa ex tempore -valmisteissa on, ettei niillä ole myyntilupaa, joten niiden tehoa ja turvallisuutta ei ole tutkittu. Annosjauheet ovat hitaita tehdä ja aikaisemmissa tutkimuksissa on havaittu lääkeaineiden tarttumista paperisiin annosjauhekuoriin. Lisäksi niiden kokonaismassa annosyksikköä kohden on suurempi kuin vastaavanvahvuisilla kapseleilla, mikä lisää vierasainekuormitusta pienillä potilailla. Tämän tutkimuksen tarkoituksena oli tutkia annosvaihtelua 4 mg:n vahvuisissa ex tempore -sotalolihydrokloridikapseleissa ja verrata niitä vastaavanlaisiin annosjauheisiin. Lisäksi haluttiin tutkia saantoa ravintoletkun läpi johdetuista kapselisisällöistä, sillä usein sairaala-apteekin valmistamia annosjauheita annostellaan ravintoletkun kautta, esimerkiksi vastasyntyneille. Keskeinen osa työtä oli kehittää sopiva HPLC -analyysimenetelmä sotalolihydrokloridille. Tämä onnistui lopulta hyvin ja aikaan saatiin tehokas ja tarkka analyysimenetelmä, joka soveltuu hyvin myös rutiinianalyyseihin 7 minuutin ajoajallaan. Kaksi kolmesta ex tempore -kapselierästä täyttivät Euroopan farmakopean vaatimukset annosvaihtelulle, kuten myös vertailuksi valmistettu annosjauhe-erä. Ravintoletkun läpi johdetut näytteet täyttivät niin ikään annosvaihtelun vaatimukset, mutta lääkeaineen määrä oli tilastollisesti merkitsevästi pienempi verrattuna kapseli- ja annosjauhenäytteisiin. Kokonaismassaltaan pienempinä ja nopeampina valmistaa kapselit ovat suositeltava vaihtoehto perinteisille annosjauheille ex tempore -lastenlääkinnässä. Tämän tutkimuksen perusteella on kuitenkin syytä ottaa huomioon mahdollisuus liian suurelle annosvaihtelulle, minkä vuoksi valmistetut erät tulisi tutkia ennen käyttöön vapauttamista. Tulevaisuudessa haasteena onkin kehittää sairaala-apteekkien mahdollisuuksia analysoida valmistamiaan lääke-eriä

Lymphatic vessels are present in human saccular intracranial aneurysms

Saccular intracranial aneurysm (sIA) rupture leads to subarachnoid haemorrhage and is preceded by chronic inflammation and atherosclerotic changes of the sIA wall. Increased lymphangiogenesis has been detected in atherosclerotic extracranial arteries and in abdominal aortic aneurysms, but the presence of lymphatic vessels in sIAs has remained unexplored. Here we studied the presence of lymphatic vessels in 36 intraoperatively resected sIAs (16 unruptured and 20 ruptured), using immunohistochemical and immunofluorescence stainings for lymphatic endothelial cell (LEC) markers. Of these LEC-markers, both extracellular and intracellular LYVE-1-, podoplanin-, VEGFR-3-, and Prox1-positive stainings were detected in 83%, 94%, 100%, and 72% of the 36 sIA walls, respectively. Lymphatic vessels were identified as ring-shaped structures positive for one or more of the LEC markers. Of the sIAs, 78% contained lymphatic vessels positive for at least one LEC marker. The presence of LECs and lymphatic vessels were associated with the number of CD68+ and CD163+ cells in the sIA walls, and with the expression of inflammation indicators such as serum amyloid A, myeloperoxidase, and cyclo-oxygenase 2, with the presence of a thrombus, and with the sIA wall rupture. Large areas of VEGFR-3 and alpha-smooth muscle actin (alpha SMA) double-positive cells were detected in medial parts of the sIA walls. Also, a few podoplanin and alpha SMA double-positive cells were discovered. In addition, LYVE-1 and CD68 double-positive cells were detected in the sIA walls and in the thrombus revealing that certain CD68+ macrophages are capable of expressing LEC markers. This study demonstrates for the first time the presence of lymphatic vessels in human sIA walls. Further studies are needed to understand the role of lymphatic vessels in the pathogenesis of sIA.Peer reviewe

Myeloperoxidase Associates With Degenerative Remodeling and Rupture of the Saccular Intracranial Aneurysm Wall

Rupture of a saccular intracranial aneurysm (sIA) is often fatal. Thus, early detection of rupture-prone sIAs is vital. Myeloperoxidase (MPO), derived mainly from neutrophils, associates with sIA rupture, and therefore its role in sIA pathogenesis warrants further studies. We analyzed MPO and its association with other histological markers in 36 (16 unruptured and 20 ruptured) sIA samples by immunohistochemistry. MPO was present in all studied sIAs, and its expression associated with wall inflammatory cell infiltrations (r = 0.50, 0.63, and 0.75, all p <0.002), degenerative remodeling (p = 0.002) and rupture (p = 0.003). MPO associated strongly with the presence of organized luminal thrombi (p <0.001), which also stained positive for MPO. Polymorphonuclear MPO+ cells were detected in the sIA walls, indicating neutrophils as MPO-source. MPO correlated strongly with accumulation of oxidized lipids (r = 0.67, p <0.001) and loss of smooth muscle cells (r = -0.68, p <0.001), suggesting that MPO is a relevant source of oxidative stress leading to cell death in the sIA wall. Furthermore, MPO associated with erythrocyte fragmentation (r = 0.74, p <0.001) and iron deposition (p = 0.041), 2 outcomes known to amplify MPO-dependent oxidative stress. Taken together, these results suggest that MPO associates with degenerative remodeling predisposing to sIA wall rupture and may serve as a biomarker of a rupture-prone sIA wall.Peer reviewe

Serum Amyloid A Is Present in Human Saccular Intracranial Aneurysm Walls and Associates With Aneurysm Rupture

Saccular intracranial aneurysm (sIA) rupture leads to a disabling subarachnoid hemorrhage. Chronic inflammation and lipid accumulation in the sIA wall contribute to wall degenerative remodeling that precedes its rupture. A better understanding of the pathobiological process is essential for improved future treatment of patients carrying sIAs. Serum amyloid A (SAA) is an acute-phase protein produced in response to acute and chronic inflammation and tissue damage. Here, we studied the presence and the potential role of SAA in 36 intraoperatively resected sIAs (16 unruptured and 20 ruptured), that had previously been studied by histology and immunohistochemistry. SAA was present in all sIAs, but the extent of immunopositivity varied greatly. SAA immunopositivity correlated with wall degeneration (p=0.028) and rupture (p=0.004), with numbers of CD163-positive and CD68-positive macrophages and CD3-positive T lymphocytes (all pPeer reviewe

Challenge of paediatric compounding to solid dosage forms sachets and hard capsules - Finnish perspective

Objectives The study evaluated the quality of compounded sachets and hard gelatine capsules and their feasibility in paediatric drug administration. Methods Commercial tablets were compounded to sachets and capsules in hospital environment, and the uniformity of content and simulated drug dose were determined. Key findings Compounded formulations were successfully obtained for a range of drug substances; dipyridamole, spironolactone, warfarin and sotalol formulations were within acceptable limits for uniformity of content, in most cases. However, some loss of drug was seen. The type and amount of excipients were found to affect uniformity of content; good conformity of capsules was obtained using lactose monohydrate as filler, whereas microcrystalline cellulose was a better choice in sachets. In capsules, content uniformity was obtained for a range of drug doses. If the drug is aimed to be administered through a nasogastric tube, solubility of the drug and excipients should be considered, as they were found to affect the simulated drug dose in administration. Conclusions Compounded sachets and capsules fulfilled the quality requirements in most cases. In compounding, the choice of excipients should be considered as they can affect conformity of the dosage form or its usability in practice. Quality assurance of compounded formulations should be taken into consideration in hospital pharmacies.Peer reviewe

Modified Lipoprotein-Derived Lipid Particles Accumulate in Human Stenotic Aortic Valves

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Population genetics and antimicrobial susceptibility of canine Campylobacter isolates collected before and after a raw feeding experiment

In recent years, increasing numbers of consumers have become interested in feeding raw food for their pet dogs as opposed to commercial dry food, in the belief of health advantages. However, raw meat and internal organs, possibly contaminated by pathogens such as Campylobacter spp., may pose a risk of transmission of zoonoses to the pet owners. Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans but C. upsaliensis has also been associated with human disease. In this study we investigated the effect of different feeding strategies on the prevalence of Campylobacter spp. in Finnish dogs. We further characterized the isolates using multilocus sequence typing (MLST), whole-genome (wg) MLST and antimicrobial susceptibility testing. Dogs were sampled before and after a feeding period consisting of commercial raw feed or dry pellet feed. Altogether 56% (20/36) of the dogs yielded at least one Campylobacter-positive fecal sample. C. upsaliensis was the major species detected from 39% of the dogs before and 30% after the feeding period. Two C. jejuni isolates were recovered, both from raw-fed dogs after the dietary regimen. The isolates represented the same genotype (ST-1326), suggesting a common infection source. However, no statistically significant correlation was found between the feeding strategies and Campylobacter spp. carriage. The global genealogy of MLST types of dog and human C. upsaliensis isolates revealed weakly clonal population structure as most STs were widely dispersed. Major antimicrobial resistance among C. upsaliensis isolates was against streptomycin (STR MIC > 4mg/l). Apart from that, all isolates were highly susceptible against the antimicrobials tested. Mutations were found in the genes rpsL or rpsL and rsmG in streptomycin resistant isolates. In conclusion, increasing trend to feed dogs with raw meat warrants more studies to evaluate the risk associated with raw feeding of pets in transmission of zoonoses to humans.Peer reviewe

STAT3 Mutation Is Associated with STAT3 Activation in CD30+ ALK− ALCL

Peripheral T-cell lymphomas (PTCL) are a heterogeneous, and often aggressive group of non-Hodgkin lymphomas. Recent advances in the molecular and genetic characterization of PTCLs have helped to delineate differences and similarities between the various subtypes, and the JAK/STAT pathway has been found to play an important oncogenic role. Here, we aimed to characterize the JAK/STAT pathway in PTCL subtypes and investigate whether the activation of the pathway correlates with the frequency of STAT gene mutations. Patient samples from AITL (n = 30), ALCL (n = 21) and PTCL-NOS (n = 12) cases were sequenced for STAT3, STAT5B, JAK1, JAK3, and RHOA mutations using amplicon sequencing and stained immunohistochemically for pSTAT3, pMAPK, and pAKT. We discovered STAT3 mutations in 13% of AITL, 13% of ALK+ ALCL, 38% of ALK− ALCL and 17% of PTCL-NOS cases. However, no STAT5B mutations were found and JAK mutations were only present in ALK- ALCL (15%). Concurrent mutations were found in all subgroups except ALK+ ALCL where STAT3 mutations were always seen alone. High pY-STAT3 expression was observed especially in AITL and ALCL samples. When studying JAK-STAT pathway mutations, pY-STAT3 expression was highest in PTCLs harboring either JAK1 or STAT3 mutations and CD30+ phenotype representing primarily ALK− ALCLs. Further investigation is needed to elucidate the molecular mechanisms of JAK-STAT pathway activation in PTCL

STAT3 Mutation Is Associated with STAT3 Activation in CD30+ ALK− ALCL

Peripheral T-cell lymphomas (PTCL) are a heterogeneous, and often aggressive group of non-Hodgkin lymphomas. Recent advances in the molecular and genetic characterization of PTCLs have helped to delineate differences and similarities between the various subtypes, and the JAK/STAT pathway has been found to play an important oncogenic role. Here, we aimed to characterize the JAK/STAT pathway in PTCL subtypes and investigate whether the activation of the pathway correlates with the frequency of STAT gene mutations. Patient samples from AITL (n = 30), ALCL (n = 21) and PTCL-NOS (n = 12) cases were sequenced for STAT3, STAT5B, JAK1, JAK3, and RHOA mutations using amplicon sequencing and stained immunohistochemically for pSTAT3, pMAPK, and pAKT. We discovered STAT3 mutations in 13% of AITL, 13% of ALK+ ALCL, 38% of ALK− ALCL and 17% of PTCL-NOS cases. However, no STAT5B mutations were found and JAK mutations were only present in ALK- ALCL (15%). Concurrent mutations were found in all subgroups except ALK+ ALCL where STAT3 mutations were always seen alone. High pY-STAT3 expression was observed especially in AITL and ALCL samples. When studying JAK-STAT pathway mutations, pY-STAT3 expression was highest in PTCLs harboring either JAK1 or STAT3 mutations and CD30+ phenotype representing primarily ALK− ALCLs. Further investigation is needed to elucidate the molecular mechanisms of JAK-STAT pathway activation in PTCL

Spatial immunoprofiling of the intratumoral and peritumoral tissue of renal cell carcinoma patients

While the abundance and phenotype of tumor-infiltrating lymphocytes are linked with clinical survival, their spatial coordination and its clinical significance remain unclear. Here, we investigated the immune profile of intratumoral and peritumoral tissue of clear cell renal cell carcinoma patients (n = 64). We trained a cell classifier to detect lymphocytes from hematoxylin and eosin stained tissue slides. Using unsupervised classification, patients were further classified into immune cold, hot and excluded topographies reflecting lymphocyte abundance and localization. The immune topography distribution was further validated with The Cancer Genome Atlas digital image dataset. We showed association between PBRM1 mutation and immune cold topography, STAG1 mutation and immune hot topography and BAP1 mutation and immune excluded topography. With quantitative multiplex immunohistochemistry we analyzed the expression of 23 lymphocyte markers in intratumoral and peritumoral tissue regions. To study spatial interactions, we developed an algorithm quantifying the proportion of adjacent immune cell pairs and their immunophenotypes. Immune excluded tumors were associated with superior overall survival (HR 0.19, p = 0.02) and less extensive metastasis. Intratumoral T cells were characterized with pronounced expression of immunological activation and exhaustion markers such as granzyme B, PD1, and LAG3. Immune cell interaction occurred most frequently in the intratumoral region and correlated with CD45RO expression. Moreover, high proportion of peritumoral CD45RO+ T cells predicted poor overall survival. In summary, intratumoral and peritumoral tissue regions represent distinct immunospatial profiles and are associated with clinicopathologic characteristics.Peer reviewe