Distribution of TRPC3 and TRPC6 in the human exocrine and endocrine pancreas.

BACKGROUND: Expression and function of TRPC3 and TRPC6 in the pancreas is a controversial topic. Investigation in human tissue is seldom. We aimed to provide here a detailed description of the distribution of TRPC3 and TRPC6 in the human exocrine and endocrine pancreas. METHODS: We collected healthy samples from cadavers (n = 4) and visceral surgery (n = 4) to investigate the respective expression profiles using immunohistochemical tracing with knockout-validated antibodies. RESULTS: TRPC3- and TRPC6-proteins were detected in different pancreatic structures including acinar cells, as well as epithelial ductal cells from intercalate, intralobular, and interlobular ducts. Respective connective tissue layers appeared unstained. Endocrine islets of Langerhans were clearly and homogenously immunolabeled by the anti-TRPC3 and anti-TRPC6 antibodies. Insular α, ß, γ, and δ cells were conclusively stained, although no secure differentiation of cell types was performed. CONCLUSIONS: Due to aforementioned antibody specificity verification, protein expression in the immunolabeled localizations can be accepted. Our study in human tissue supports previous investigations especially with respect to acinar and insular α and ß cells, while other localizations are here reported for the first time to express TRPC3 and TRPC6, ultimately warranting further research.

The Attallah screw: Where safety meets robustness in posterior subaxial cervical instrumentation.

Posterior fixation of the subaxial cervical spine (SCS) commonly relies on the application of lateral mass screws (LMS), with pedicle screws being a less prevalent alternative. The present study provides another option: A recently introduced novel approach, the Attallah screw, intended to ensure a safety profile comparable to that of LMS, combined with a strength profile similar to that of pedicle screws. The focus of the present study is the comparative analysis of peak insertion torques for these three screw types. Employing standard surgical techniques and instruments, Attallah screws were scheduled for insertion on the right side of the SCS in 15 cadavers, pedicle screws on the left side in 8 cadavers, and LMS on the left side in the remaining 7 cadavers. The peak insertion torque was recorded using an electronic torque screwdriver. The results revealed that the peak insertion torques were similar in the pedicle and the Attallah screw at C3, C4 and C7, but differed at C5 (mean ± SD; pedicle, 79.5±19.6 cNm; Attallah, 56.7±18.5 cNm; P=0.029) and C6 (pedicle, 85.4±28.7 cNm; Attallah, 49.8±17.9 cNm; P=0.004) in favor of the superior pedicle screw measurements. The peak insertion torques of the pedicle screw were superior to the corresponding data from the LMS from C4 to C7. By contrast, the peak insertion torques of the Attallah screw were only superior to those of the LMS at C7 (Attallah, 69.5±24.5 cNm; lateral mass, 40.5±21.4 cNm; P=0.030), although similar trends were observed at the other cervical levels. On the whole, the findings presented herein indicate the level-dependent superior robustness of the Attallah screw as a posterior cervical fixation method compared to the LMS. However, from a biomechanical perspective, the pedicle screw remains the preeminent choice for fixation within the C5-C6 range.

TRPC3 Is Downregulated in Primary Hyperparathyroidism.

Transient receptor potential canonical sub-family channel 3 (TRPC3) is considered to play a critical role in calcium homeostasis. However, there are no established findings in this respect with regard to TRPC6. Although the parathyroid gland is a crucial organ in calcium household regulation, little is known about the protein distribution of TRPC channels-especially TRPC3 and TRPC6-in this organ. Our aim was therefore to investigate the protein expression profile of TRPC3 and TRPC6 in healthy and diseased human parathyroid glands. Surgery samples from patients with healthy parathyroid glands and from patients suffering from primary hyperparathyroidism (pHPT) were investigated by immunohistochemistry using knockout-validated antibodies against TRPC3 and TRPC6. A software-based analysis similar to an H-score was performed. For the first time, to our knowledge, TRPC3 and TRPC6 protein expression is described here in the parathyroid glands. It is found in both chief and oxyphilic cells. Furthermore, the TRPC3 staining score in diseased tissue (pHPT) was statistically significantly lower than that in healthy tissue. In conclusion, TRPC3 and TRPC6 proteins are expressed in the human parathyroid gland. Furthermore, there is strong evidence indicating that TRPC3 plays a role in pHPT and subsequently in parathyroid hormone secretion regulation. These findings ultimately require further research in order to not only confirm our results but also to further investigate the relevance of these channels and, in particular, that of TRPC3 in the aforementioned physiological functions and pathophysiological conditions.

Long-Term Astigmatism after Intraocular Pressure Sensor Implantation and Nonpenetrating Glaucoma Surgery - EYEMATE-SC Trial.

PURPOSE: To investigate the long-term astigmatism after combined non-penetrating glaucoma surgery (NPGS) and implantation of the first miniaturized suprachoroidal intraocular pressure (IOP) sensor EYEMATE-SC. SETTING: The study was conducted in five medical centers in two different countries. DESIGN: Retrospective multicenter clinical study. METHODS: Astigmatism of patients instrumented with the EYEMATE-SC IOP sensor was assessed over a follow-up period of three years. Refraction and corrected distance visual acuity (CDVA) were obtained preoperatively, after 6 months, 1, 2, and 3 years. A canaloplasty-operated patient cohort served as control. Astigmatism was evaluated using 3-dimensional power vector analysis involving the spherical equivalent M, and the Jackson crossed cylinder projections J0 and J45. Exclusion criteria included neovascular and angle-closure glaucoma, myopia, axial length outside 22 to 26 mm, other ocular diseases, prior glaucoma surgery, other ocular surgery within 6 months (cataract surgery within 3 months) prior to NPGS, serious generalized conditions, and other active medical head/neck implants. RESULTS: Multivariate analysis indicated no changes in astigmatism along the observation period in both the EYEMATE-SC (n = 24) and the canaloplasty (n = 24) group (P > 0.05 or nonsignificant after Bonferroni correction). Astigmatism was unchanged between the EYEMATE-SC and the canaloplasty group at all time points (P > 0.05). CDVA didn't change along the observation period of three years in each of both groups (P > 0.05). CONCLUSIONS: Despite its suprachoroidal localization, the present study indicates that the miniaturized EYEMATE-SC IOP sensor doesn't negatively affect the long-term astigmatism after combined implantation with NPGS.

Postmortem sympathomimetic iris excitability.

BACKGROUND: A reliable estimation of time since death can be important for the law enforcement authorities. The compound method encompassing supravital reactions such as the chemical excitability of the iris can be used to further narrow intervals estimated by temperature-based methods. Postmortem iris excitability was mostly assessed by parasympatholytic or parasympathomimetic substances. Little is known regarding sympathomimetic agents. The present study aims to describe the postmortem iris excitability using the sympathomimetic drug phenylephrine. METHODS: Cadavers were included after body donors gave written informed consent during lifetime. Exclusion criteria were known eye disease, or a postmortem interval exceeding 26 hours. A pupillometer with a minimum measurement range of 0.5 mm was used to determine the horizontal pupil diameter before and 20 minutes after the application of phenylephrine. Increase in pupil diameter was labeled as positive reaction, unchanged pupil diameter was labeled as negative reaction, and decrease in pupil diameter was labeled as paradox reaction. RESULTS: 30 eyes from 16 cadavers (median age = 80.0; 9 males, 7 females) were examined. Initial pupil size was in median 3.5 mm (interquartile range [IQR]: 3.0-4.5 mm) and progressed to 4.0 mm (IQR: 3.5-5.0 mm) 20 minutes after drug instillation. The achieved pupil diameter difference comprised in median 0.5 mm (IQR: 0.0-1.0 mm). A positive reaction was observed in 21 cases. Negative reactions were observed in 5 cases and paradox reactions in 4 cases. Overall, there was a statistically significant difference in diameter between the initial and the reactive pupil (P = 0.0002). CONCLUSION: Although relatively rarely used, sympathomimetic drugs seem to be eligible for chemical postmortem iris excitability. Currently, assessment of postmortem iris excitability usually only involves parasympatholytic and parasympathomimetic agents. The findings of the present study give a hint that the application of a third agent with a sympathomimetic mechanism of action could provide additional information. Further studies assessing such a triple approach in the compound method in comparison with the current gold standard for estimation of time since death are mandatory to ensure reliable results.

New insights in the renal distribution profile of TRPC3 - Of mice and men.

Several reports previously investigated the Transient Receptor Potential Canonical subfamily channel 3 (TRPC3) in the kidney. However, most of the conclusions are based on animal samples or cell cultures leaving the door open for human tissue investigations. Moreover, results often disagreed among investigators. Histological description is lacking since most of these studies focused on functional aspects. Nevertheless, the same reports highlighted the potential key-role of TRPC3 in renal disorders. Hence, our interest to investigate the localization of TRPC3 in human kidneys. For this purpose, both healthy mouse and human kidney samples that were originated from tumor nephrectomies have been prepared for immunohistochemical staining using a knockout-validated antibody. A blocking peptide was used to confirm antibody specificity. A normalized weighted diaminobenzidine (DAB) area score between 0 and 3 comparable to a pixelwise H-score was established and employed for semiquantitative analysis. Altogether, our results suggest that glomeruli only express little TRPC3 compared to several segments of the tubular system. Cortical and medullary proximal tubules are stained, although intracortical differences in staining exist in mice. Intermediate tubules, however, are only weakly stained. The distal tubule was studied in three localizations and staining was marked although slightly varying throughout the different subsegments. Finally, the collecting duct was also immunolabeled in both human and mouse tissue. We therefore provide evidence that TRPC3 is expressed in various localizations of both human and mouse samples. We verify results of previous studies and propose until now undescribed localizations of TRPC3 in the mouse but especially and of greater interest in the human kidney. We thereby not only support the translational concept of the TRPC3 channel as key-player in physiology and pathophysiology of the human kidney but also present new potential targets to functional analysis.

Immunohistochemistry Reveals TRPC Channels in the Human Hearing Organ-A Novel CT-Guided Approach to the Cochlea.

TRPC channels are critical players in cochlear hair cells and sensory neurons, as demonstrated in animal experiments. However, evidence for TRPC expression in the human cochlea is still lacking. This reflects the logistic and practical difficulties in obtaining human cochleae. The purpose of this study was to detect TRPC6, TRPC5 and TRPC3 in the human cochlea. Temporal bone pairs were excised from ten body donors, and the inner ear was first assessed based on computed tomography scans. Decalcification was then performed using 20% EDTA solutions. Immunohistochemistry with knockout-tested antibodies followed. The organ of Corti, the stria vascularis, the spiral lamina, spiral ganglion neurons and cochlear nerves were specifically stained. This unique report of TRPC channels in the human cochlea supports the hypothesis of the potentially critical role of TRPC channels in human cochlear health and disease which has been suggested in previous rodent experiments.

Establishing an experimental Pseudomonas aeruginosa keratitis model in mice - Challenges and solutions.

BACKGROUND: With the ongoing increase in antimicrobial resistances seen in bacterial isolates causing a keratitis in humans, animal models have become an important tool to study new antimicrobial therapies. Nevertheless, the establishment of experimental keratitis is difficult. Here, we discuss the impact of different arrangements, including animal age, bacterial strain and dose as well as epithelium removal on the outcome of experimental keratitis. We therefore present the methods and results of our establishing experiments. METHODS: Bacterial load determination and flow cytometry were performed using eye homogenate gained from a 72 h lasting murine Pseudomonas aeruginosa keratitis model. Additionally, the intensity of the infection was scored from 0 to 5, the mice weighed, and blood immune cells counted. RESULTS: We found that older C57BL/6 N mice (8-11 months) are more susceptible to develop a keratitis than younger mice (5-6 weeks). Epithelium removal has no major impact on infectivity and disease progression in aged mice. P. aeruginosa exoU+ strains, such as PA54, should preferentially be used and highly concentrated (∼ 5 ×107 colony forming units CFU). Establishing an infection with the exoU- PAO1 derivative DSM 19880 was not possible. CONCLUSIONS: We present a replicable method to achieve a successful experimental P. aeruginosa keratitis in C57BL/6 N mice that is sustained or aggravated over the observation period of 3 days in 80 % of all animals tested. Our work is of particular interest to all researchers planning the establishment of such experimental models. We show some key aspects that can simplify and quicken the procedure, ultimately saving costs and animal life.

Comparison of the maximum possible lengths of insertable screws in the Subaxial Cervical Spine.

BACKGROUND: Lateral mass screws are the most commonly used fixation technique in the Subaxial Cervical Spine (SCS), their main advantages being that they are easy to insert and safe in their application. Pedicle screws are significantly longer, are quite challenging to insert in most settings and are accompanied by the risk of serious complications such as vascular and neural injuries. We have therefore developed a new technique, which permits safe insertion of long screws in the SCS. METHODS: A radiological evaluation was carried out to determine the maximum possible insertable screw length in the SCS when using the following techniques: pedicle, lateral mass (Magerl's) and "Attallah" screws. Scans of 66 cervical spines were analyzed to determine the maximum possible screw lengths of all three screw insertion techniques, based on the standard description through the vertebrae from C3 to C7. RESULTS: The maximum possible length of the Attallah screw of 20.7 ± 2.5 mm (mean value ± SD) is only 2.4 mm shorter than the pedicle screw (23.1 ± 1.8 mm) along the SCS. The lateral mass screw is with 10.2 ± 1.3 mm full 12.9 mm shorter than the pedicle screw. CONCLUSIONS: The maximum possible length of the Attallah screw is close to that of the pedicle screw and significantly greater than that of the lateral mass screw. We provide a fixation method comparable to the pedicle screw in its strength and to the lateral mass screw in its safety.

TRPC Channels in the Physiology and Pathophysiology of the Renal Tubular System: What Do We Know?

The study of transient receptor potential (TRP) channels has dramatically increased during the past few years. TRP channels function as sensors and effectors in the cellular adaptation to environmental changes. Here, we review literature investigating the physiological and pathophysiological roles of TRPC channels in the renal tubular system with a focus on TRPC3 and TRPC6. TRPC3 plays a key role in Ca2+ homeostasis and is involved in transcellular Ca2+ reabsorption in the proximal tubule and the collecting duct. TRPC3 also conveys the osmosensitivity of principal cells of the collecting duct and is implicated in vasopressin-induced membrane translocation of AQP-2. Autosomal dominant polycystic kidney disease (ADPKD) can often be attributed to mutations of the PKD2 gene. TRPC3 is supposed to have a detrimental role in ADPKD-like conditions. The tubule-specific physiological functions of TRPC6 have not yet been entirely elucidated. Its pathophysiological role in ischemia-reperfusion injuries is a subject of debate. However, TRPC6 seems to be involved in tumorigenesis of renal cell carcinoma. In summary, TRPC channels are relevant in multiples conditions of the renal tubular system. There is a need to further elucidate their pathophysiology to better understand certain renal disorders and ultimately create new therapeutic targets to improve patient care.

Radiological analyses of the dimensions of the pedicle and dorsal part of the transverse process of subaxial vertebrae in the context of cervical spine surgery.

BACKGROUND: Knowledge of the anatomical dimensions of distinct areas of the vertebral bodies and vertebral arches of the subaxial cervical spine are indispensable in the planning of osteosynthesis. The minute dimensions and complex anatomical orientation of the posterior vertebral arch structures in the subaxial spine render the insertion of screws - where needed - a challenging procedure. The pedicle option carries the risk of serious complications while the alternative of lateral mass only permits the insertion of short screws. OBJECTIVE: The transverse process of the subaxial vertebrae offers unique possibilities on all counts and seems quite comparable in its dimensions with the pedicle. To our knowledge it has not been used previously for the insertion of screws in the subaxial spine. METHODS: Therefore, the scans of 66 cervical spines were analysed for distinct lines of both structures. RESULTS: The widths and lengths of the dorsal part of the transverse processes and of the pedicles are similar between both sides. Clear differences between females and males could be observed. The widths of both structures were closer to each other in C3 than in C4 to C7, while the lengths derived the most in C7. CONCLUSION: The dorsal part of the transverse process might be suitable for the insertion of screws to stabilize the vertebral arch of the subaxial cervical spine. Gender adaptation might be required.

Lesions in the lungs of fatal corona virus disease Covid-19.

The corona virus outbreak in Wuhan, China, at the end of 2019 has rapidly evolved into a pandemic which is still virulent in many countries. An infection with SARS-CoV-2 can lead to corona virus disease (Covid-19). This paper presents an overview of the knowledge gained so far with regard to histopathological lung lesions in fatal courses of Covid-19. The main findings were diffuse alveolar damage and micro-angiopathies. These included the development of hyaline membranes, thrombi, endothelial inflammation, haemorrhages and angiogenesis. Overall, the vessel lesions seemed to be more lethal than the diffuse alveolar damage. There was obvious hyperreactivity and hyperinflammation of the cellular immune system. An expanded T-cell memory may explain the increased risk of a severe course in the elderly.