Human pluripotent stem cell-derived cells endogenously expressing follicle-stimulating hormone receptors: modeling the function of an inactivating receptor mutation.

Follicle-stimulating hormone (FSH) is crucial in the development and regulation of reproductive functions. The actions of human FSH and its receptor (FSHR) and mutations therein have mainly been studied using in vivo models, primary cells, cancer cells and cell lines ectopically expressing the FSHR. To allow studies of endogenous FSHR function in vitro, we differentiated FSHR-expressing cells from human pluripotent stem cells. FSH stimulation of the wild-type (WT), but not the inactivating Finnish founder mutant (A189V) receptor, activated the canonical cyclic adenosine monophosphate (cAMP)-dependent signaling pathway and downstream mediators. To investigate protein-protein interaction partners of FSHR at resting state and upon FSH stimulation, we expressed FSHR in HEK293 cells followed by affinity purification mass spectrometry analyses. We found 19 specific high-confidence interacting proteins for WT FSHR and 14 for A189V FSHR, several of which have been linked to infertility. Interestingly, while only WT FSHR interacted with FSH, insulin-like growth factor 1 receptor (IGF1R), for example, interacted with both WT and A189V FSHR upon FSH stimulation. In conclusion, our protocol allows detailed studies of FSH action and disease modeling in human cells endogenously expressing FSHR.

Prescribing for off-label use and unauthorized medicines in three paediatric wards in Finland, the status before and after the European Union Paediatric Regulation.

WHAT IS KNOWN AND OBJECTIVE: The European Paediatric Regulation aims to reduce off-label use of medicines in paediatric pharmacotherapy. Prescribing for off-label use and unauthorized medicines was common in the paediatric wards of the Kuopio University Hospital in 2001. To evaluate the possible impact of the Regulation on the prevalence and the frequency on such prescribing, we repeated the study in 2011 as it was conducted 10 years earlier. METHODS: In this prospective study, the prescriptions for patients below 18 years of age were reviewed during a 2-week period in each of the three wards; neonatal intensive care unit, general paediatric ward and paediatric surgical ward in April and May 2011. The medicine's authorizing status of all prescriptions was determined according to the approved summary of product characteristics valid during the study in Finland. Data concerning unauthorized medicines were also recorded and classified. RESULTS: Out of the entire study population of 123 patients, 119 received a total of 1054 prescriptions in 2011. The proportion of patients with at least one prescription for off-label use or for an unauthorized medicine was significantly higher, 79% (n = 97) in 2011, compared to 58% in 2001 (P < 0·001). For newborns, significantly more prescriptions were for off-label use in 2011 than in 2001 (51% vs. 22%; P < 0·001). The proportion of prescriptions for unauthorized medicines was significantly higher in children below 2 years of age than in older children in both years (21% vs. 5% in 2011 and 24% vs. 3% in 2001, P < 0·001). WHAT IS NEW AND CONCLUSION: The prescribing for off-label use and unauthorized medicines was more prevalent in 2011 than in 2001. This indicates that the recent legislation has had only minor or no impact on the authorizing status of medicines commonly used in paediatric inpatients in specialized care.

Parental reporting of adverse drug events and other drug-related problems in children in Finland.

PURPOSE: Drug-related problems (DRPs) are common in paediatric pharmacotherapy, but few studies describe them from the parents' perspective. In the present survey, we have investigated the lifetime prevalence and type of DRPs in children in Finland. METHODS: This was a population-based survey of a random sample of 6,000 children below 12 years of age in 2007. A questionnaire was sent to their parents. The final response rate was 67% with a study population of 4,032. The main outcome measure was the lifetime prevalence and type of DRPs. RESULTS: The lifetime prevalence of DRPs was 21% (95% CI 20-22). The most common (82%) of the 1,346 reported DRPs were adverse drug events (ADEs). The prevalence of ADEs was 17% (95% CI 16-19), that of other DRPs 5.2% (95% CI 4.5-5.9). The prevalence of serious ADEs was 0.4% and that of unexpected ADEs was 0.8%. The most common system involved in the ADEs was the gastrointestinal tract, comprising 34% of the 1,106 ADEs. The most common of the 240 other DRPs were problems with the administration and dosing of medicine (86%). Overall, 64% of DRPs were related to anti-infectives. CONCLUSIONS: One fifth of the Finnish children below 12 years of age had experienced DRPs. Appropriate counselling, including possible adverse drug reactions and dosing directions, is important for parents and children at both the prescribing and dispensing of medicines for paediatric patients. Reporting of any suspected serious or unexpected ADEs is an essential part of efficient pharmacovigilance in paediatrics.

Expression of CD147 and monocarboxylate transporters MCT1, MCT2 and MCT4 in porcine small intestine and colon.

Lactate, formed mainly in the stomach and small intestines, and short-chain fatty acids (SCFAs) formed in the colon, are ionised and require transporter proteins such as monocarboxylate transporters (MCTs) for absorption. The amounts of MCT1, MCT2, MCT4 and CD147, an ancillary protein for MCT1 and MCT4, were measured by immunoblotting the small intestine and colon of 40 pigs (Landrace, Yorkshire and LandracexYorkshire). MCT1 and MCT4 were found in both small intestine and colon, but MCT2 only in the small intestine. In both small intestine and colon, Yorkshire pigs had more CD147 than Landrace pigs, while no interbreed differences were found in MCT isoforms. Since CD147 is essential for the activity of MCT1 and MCT4, the breed difference suggests that MCT activity is higher in Yorkshire than in Landrace pigs. The absence of MCT2 in the colon suggests that it is mainly a lactate transporter, while MCT1 and MCT4 facilitate the transport of both lactate and SCFA.

The inducible form of heat shock protein 70 in the serum, colon and small intestine of the pig: comparison to conventional stress markers.

Modern rearing conditions may cause stress to pigs. At the cellular level all animals respond to stress by synthesizing heat shock proteins (HSP), which protect cells from injury. The objective of this study was to examine the concentrations of stress-inducible HSP72 in porcine small intestine and colon, known to be stress sensitive tissues, and to compare the findings with HSP72 concentrations in serum and with conventional markers of stress, namely blood lactate and serum cortisol, glucose, free fatty acids and acute phase proteins. HSP72 in the colon correlated with serum HSP72 but there was a negative correlation with carcass weight (growth). The results suggest that the colon may be a significant source of serum HSP72, the concentration of which may reflect changes in the permeability of intestinal epithelium due to stressors, such as transport and handling.

Training of medical staff positively influences postoperative pain management at home in children.

The aim of this study was to describe how parents manage their child's postoperative pain at home following day-case surgery. The incidence of pain, different analgesics used and problems related to administering medications were the main interests of the study. A postal questionnaire was sent to the parents of 275 children who were under 8 years of age and had undergone an ear, nose and throat (ENT) day-case operation. The questionnaire was sent to the parents a week after discharge from hospital. Altogether, the parents of 227 children answered the questionnaire (response rate 83%). The study was divided into two phases (preintervention and postintervention), and incorporated a training program for doctors and nurses between these two phases. The training program aimed to improve the treatment practices of postoperative pain in children. Seventy-eight per cent of the children in the preintervention study and 75% in the postintervention study experienced at least mild pain after discharge. The training program for doctors and nurses affected the home treatment practices of postoperative pain. The proportion of parents treating their children increased from 68% to 80% after the training program (p = 0.028). Many parents faced problems while treating their children; for example, 19% (n = 30) of the children refused to take their medicine, and suppositories were regarded to be an especially unpleasant dosage form. However, no serious adverse effects were reported. We conclude that due to the pain experienced at home by the great majority of children following day-case ENT operations, parents need information on how to manage their child's pain. A training program for doctors and nurses can improve the treatment of children's pain even at home. Since some children dislike suppositories, it would be worth considering the use of small tablets or mixtures instead.

The effects of a hospital staff training program on the treatment practices of postoperative pain in children under 8 years.

The widespread use of day case surgery for minor operations in children places increased emphasis on home care, parental responsibility and attention to suitable analgesia. This study evaluated a training program for doctors and nurses on postoperative analgesia in children by following up parental perceptions of pain relief after discharge. The main objective of our study was to determine the effects of a training program on treatment practices in the hospital and on the home care instructions provided. The study was carried out in two phases. A questionnaire was sent to parents of children aged under 8 years who had undergone a minor otorhinolaryngological operation (tonsillectomy, adenoidectomy, irrigation of maxillary sinus, insertion of grommets, or paracentesis) at the Day Case Unit in Kuopio University Hospital. The parents of 115 children returned the questionnaire in the preintervention study and those of 112 children in the postintervention study (response rates: 79% and 86% respectively). The training program for personnel (intervention) was organized between the two studies. It started with a one-hour lecture on postoperative pain and pain treatment in children, and the lecture was followed by two weeks of bedside teaching. The overall standard of pain treatment in the hospital was good. Most of the children had no pain or only mild pain on discharge (93% before the intervention and 90% after the intervention). The intervention had no effect on the likelihood of children receiving pain medication. Noticeable changes occurred in the selection of pain medicines and dosage forms, and in the increased number of home care instructions provided for parents. We conclude that the training program for both professional groups (doctors and nurses) can be beneficial in changing the treatment practice (especially the use of medicines and dosage forms). Such training programs also make it possible to emphasized the importance of proper instructions for home care and thus to improve the quality of information provided for parents.

Distribution of monocarboxylate transporter isoforms MCT1, MCT2 and MCT4 in porcine muscles.

AIM: Monocarboxylate transporters (MCT), which cotransport lactate anions and protons across cell membranes, are important for regulation of muscle pH. We measured amounts of MCT1, MCT2 and MCT4 by immunoblotting in five different porcine muscles, to study MCT-isoform distribution both in oxidative and highly glycolytic muscles. METHODS: Samples from the longissimus dorsi, gluteus superficialis, semimembranosus, infraspinatus and masseter were taken from 18 slaughtered pigs. RESULTS: Oxidative capacity, estimated on the basis of the activities of lactate dehydrogenase (LDH), citrate synthase (CS) and 3-OH-acyl-CoA dehydrogenase (HAD), was highest in the infraspinatus and masseter, and was very low in the gluteus, semimembranosus and longissimus dorsi. In all muscles, the amount of MCT1 was small but variable. The amount of MCT2 was more abundant in the glycolytic than in the oxidative muscles, while MCT4 was found in equal amounts in all muscles. MCT2, but not MCT4, correlated negatively with CS and HAD. CONCLUSIONS: The results together with measured concentrations of lactate suggest that MCT2 may function as the housekeeping lactate transporter, preventing acidification especially in highly glycolytic muscles in which the capacity to oxidize lactate is low. The results also support the view that, as in other species, MCT4 would be important at high lactate concentrations that occur during stress.