Structural heart defects associated with ETB mutation, a cause of Hirschsprung disease

Background HSCR, a colonic neurocristopathy affecting 1/5000 births, is suggested to associate with cardiac septal defects and conotruncal malformations. However, we question subtle cardiac changes maybe more commonly present due to multi-regulations by HSCR candidate genes, in this instance, ETB. To investigate, we compared the cardiac morphology and quantitative measurements of sl/sl rat to those of the control group. Methods Eleven neonatal rats were generated from heterozygote (ETB+/−) crossbreeding. Age and bodyweight were recorded at time of sacrifice. Diffusion-staining protocols with 1.5% iodine solution was completed prior to micro-CT scanning. All rats were scanned using an in vivo micro-CT scanner, Caliper Quantum FX, followed by two quality-control scans using a custom-built ex vivo micro-CT system. All scans were reviewed for gross cardiac dysmorphology. Micro-CT data were segmented semi-automatically post-NLM filtering for: whole-heart, LV, RV, LA, RA, and aortic arch. Measurements were taken with Drishti. Following image analysis, PCR genotyping of rats was performed: five sl/sl rats, three wildtype, and three heterozygotes. Statistical comparisons on organ volume, growth rate, and organ volume/bodyweight ratios were made between sl/sl and the control group. Results Cardiac morphology and constituents were preserved. However, significant volumetric reductions were recorded in sl/sl rats with respect to the control: whole heart (38.70%, p value = 0.02); LV (41.22%, p value = 0.01), RV (46.15%, p value = 0.02), LA (44.93%, p value = 0.06), and RA (39.49%, p value = 0.02). Consistent trend was observed in growth rate (~ 20%) and organ-volume/bodyweight ratios (~ 25%). On the contrary, measurements on aortic arch demonstrated no significant difference among the two groups. Conclusion Despite the presence of normal morphology, significant cardiac growth retardation was detected in sl/sl rat, supporting the likely association of cardiac anomalies with HSCR, at least in ETB−/− subtype. Structural reduction was likely due to a combination of failure to thrive from enteric dysfunction, alterations to CaNCC colonization, and importantly coronary hypoperfusion from elevated ET-1/ETA-mediated hypervasoconstriction. Little correlation was detected between aortic arch development and sl/sl rat, supporting minor ETB role in large vessels. Although further clinical study is warranted, HSCR patients may likely require cardiac assessment in view of potential congenital cardiac defects.

The advantages of abdominal compression with shallow breathing during left-sided postmastectomy radiotherapy by Helical TomoTherapy

Background Left-sided post-mastectomy radiotherapy (PMRT) certainly precedes some radiation dose to the cardiopulmonary organs causing many side effects. To reduce the cardiopulmonary dose, we created a new option of the breathing adapted technique by using abdominal compression applied with a patient in deep inspiration phase utilizing shallow breathing. This study aimed to compare the use of abdominal compression with shallow breathing (ACSB) with the free breathing (FB) technique in the left-sided PMRT. Materials and methods Twenty left-sided breast cancer patients scheduled for PMRT were enrolled. CT simulation was performed with ACSB and FB technique in each patient. All treatment plans were created on a TomoTherapy planning station. The target volume and dose, cardiopulmonary organ volume and dose were analyzed. A linear correlation between cardiopulmonary organ volumes and doses were also tested. Results Regarding the target volumes and dose coverage, there were no significant differences between ACSB and FB technique. For organs at risk, using ACSB resulted in a significant decrease in mean (9.17 vs 9.81 Gy, p<0.0001) and maximum heart dose (43.79 vs 45.45 Gy, p = 0.0144) along with significant reductions in most of the evaluated volumetric parameters. LAD doses were also significantly reduced by ACSB with mean dose 19.24 vs 21.85 Gy (p = 0.0036) and the dose to 2% of the volume (D2%) 34.46 vs 37.33 Gy (p = 0.0174) for ACSB and FB technique, respectively. On the contrary, the lung dose metrics did not show any differences except the mean V5 of ipsilateral lung. The positive correlations were found between increasing the whole lung volume and mean heart dose (p = 0.05) as well as mean LAD dose (p = 0.041) reduction. Conclusions The ACSB technique significantly reduced the cardiac dose compared with the FB technique in left-sided PMRT treated by Helical TomoTherapy. Our technique is uncomplicated, well-tolerated, and can be applied in limited resource center.

Brain size reductions associated with endothelin B receptor mutation, a cause of Hirschsprung’s disease

Background ETB has been reported to regulate neurogenesis and vasoregulation in foetal development. Its dysfunction was known to cause HSCR, an aganglionic colonic disorder with syndromic forms reported to associate with both small heads and developmental delay. We therefore asked, "is CNS maldevelopment a more general feature of ETB mutation?" To investigate, we reviewed the micro-CT scans of an ETB−/− model animal, sl/sl rat, and quantitatively evaluated the structural changes of its brain constituents. Methods Eleven neonatal rats generated from ETB+/− cross breeding were sacrificed. Micro-CT scans were completed following 1.5% iodine-staining protocols. All scans were reviewed for morphological changes. Selected organs were segmented semi-automatically post-NLM filtering: TBr, T-CC, T-CP, OB, Med, Cer, Pit, and S&I Col. Volumetric measurements were made using Drishti rendering software. Rat genotyping was completed following analysis. Statistical comparisons on organ volume, organ growth rate, and organ volume/bodyweight ratios were made between sl/sl and the control groups based on autosomal recessive inheritance. One-way ANOVA was also performed to evaluate potential dose-dependent effect. Results sl/sl rat has 16.32% lower body weight with 3.53% lower growth rate than the control group. Gross intracranial morphology was preserved in sl/sl rats. However, significant volumetric reduction of 20.33% was detected in TBr; similar reductions were extended to the measurements of T-CC, T-CP, OB, Med, and Pit. Consistently, lower brain and selected constituent growth rates were detected in sl/sl rat, ranging from 6.21% to 11.51% reduction. Lower organ volume/bodyweight ratio was detected in sl/sl rats, reflecting disproportional neural changes with respect to body size. No consistent linear relationships exist between ETB copies and intracranial organ size or growth rates. Conclusion Although ETB−/− mutant has a normal CNS morphology, significant size reductions in brain and constituents were detected. These structural changes likely arise from a combination of factors secondary to dysfunctional ET-1/ET-3/ETB signalling, including global growth impairment from HSCR-induced malnutrition and dysregulations in the neurogenesis, angiogenesis, and cerebral vascular control. These changes have important clinical implications, such as autonomic dysfunction or intellectual delay. Although further human study is warranted, our study suggested comprehensive managements are required for HSCR patients, at least in ETB−/− subtype.

New Insights into YES-Associated Protein Signaling Pathways in Hematological Malignancies: Diagnostic and Therapeutic Challenges

The Hippo/YES-associated protein (YAP) signaling pathway is a cell survival and proliferation-control system with its main activity that of regulating cell growth and organ volume. YAP operates as a transcriptional coactivator in regulating the onset, progression, and treatment response in numerous human tumors. Moreover, there is evidence suggesting the involvement of YAP in the control of the hematopoietic system, in physiological conditions rather than in hematological diseases. Nevertheless, several reports have proposed that the effects of YAP in tumor cells are cell-dependent and cell-type-determined, even if YAP usually interrelates with extracellular signaling to stimulate the onset and progression of tumors. In the present review, we report the most recent findings in the literature on the relationship between the YAP system and hematological neoplasms. Moreover, we evaluate the possible therapeutic use of the modulation of the YAP system in the treatment of malignancies. Given the effects of the YAP system in immunosurveillance, tumorigenesis, and chemoresistance, further studies on interactions between the YAP system and hematological malignancies will offer very relevant information for the targeting of these diseases employing YAP modifiers alone or in combination with chemotherapy drugs.

The Advantages of Abdominal Compression with Shallow Breathing During Left-sided Postmastectomy Radiotherapy by Helical Tomotherapy

BackgroundLeft-sided post-mastectomy radiotherapy (PMRT) certainly precedes some radiation dose to the cardiopulmonary organs causing many side effects. To reduce the cardiopulmonary dose, we created a new option of the breathing adapted technique by using abdominal compression applied with a patient in deep inspiration phase utilizing shallow breathing. This study aimed to compare the use of abdominal compression with shallow breathing (ACSB) with the free breathing (FB) technique in the left-sided PMRT.MethodsTwenty left-sided breast cancer patients scheduled for PMRT were enrolled. CT simulation was performed with ACSB and FB technique in each patient. All treatment plans were created on a TomoTherapy planning station. The target volume and dose, cardiopulmonary organ volume and dose were analyzed. A linear correlation between cardiopulmonary organ volumes and doses were also tested.ResultsRegarding the target volumes and dose coverage, there were no significant differences between ACSB and FB technique. For organs at risk, using ACSB resulted in a significant decrease in mean (9.17 vs 9.81 Gy, p < 0.0001) and maximum heart dose (43.79 vs 45.45 Gy, p = 0.0144) along with significant reductions in most of the evaluated volumetric parameters. LAD doses were also significantly reduced by ACSB with mean dose 19.24 vs 21.85 Gy (p = 0.0036) and the dose to 2% of the volume (D2%) 34.46 vs 37.33 Gy (p = 0.0174) for ACSB and FB technique, respectively. On the contrary, the lung dose metrics did not show any differences except the mean V5 of ipsilateral lung. The positive correlations were found between increasing the whole lung volume and mean heart dose (p = 0.05) as well as mean LAD dose (p = 0.041) reduction.ConclusionsThe ACSB technique significantly reduced the cardiac dose compared with the FB technique in left-sided PMRT treated by Helical Tomotherapy. Our technique is uncomplicated, well-tolerated, and can be applied in daily clinical practice.Trial registrationThai Clinical Trials Registry (TCTR), TCTR20201215008. Registered 14 December 2020 - Retrospectively registered, http:// http://www.clinicaltrials.in.th/ TCTR20201215008

Necrosis and perforation of the stomach in newborn babies and infants

The Objective of the study was to identify the clinical features of newborns and infants with perforation of the stomach, and to justify the possibility of organ-preserving operations even with extensive gastric necrosis.Methods and Materials. The results of treatment of 32 newborns with stomach perforation was analyzed: not only the risk factors that cause this condition, but diagnostic methods and variants of surgical treatment. All patients with extensive necrosis of the stomach wall underwent an atypical resection within healthy tissues, a gastric «tube» was formed on the drainage probe with a significant decrease of organ volume. In cases of the local damage of the gastric wall, the perforated area was sutured after the excision of the edges of the defect.Results. Mortality rate was 36.5 % (n = 12). The cause of death in 5 children (15 %), in 3 to 8 days after surgery, was multiple organ failure syndrome. In 7 patients (22 %), a fatal outcome occurred due to the severe post-intensive care syndrome at the age of 3 to 12 months of life.Conclusion. The mechanism of perforations of the stomach in newborns and infants is multifactorial. All children with stomach perforation need preoperative preparation. The operation of choice for the stomach perforation is an organpreserving surgery. The function of the stomach is restored in all children after extensive resection of the stomach.

Larger organ size caused by obesity is a mechanism for higher cancer risk

Obesity increases significantly cancer risk in various organs. Although this has been recognized for decades, the mechanism through which this happens has never been explained. Here, we show that obese people (BMI ≥30) have on average 55% (95%CI: 46%-66%), 68% (95%CI: 59%-76%), and 39% (95%CI: 29%-49%) larger kidneys, liver, and pancreas, respectively. We also find a significant linear relationship between the increase in organ volume and the increase in cancer risk (P-value<10−12). These results provide a mechanism explaining why obese individuals have higher cancer risk in several organs: the larger the organ volume the more cells at risk of becoming cancerous. These findings are important for a better understanding of the effects that obesity has on cancer risk and, more generally, for the development of better preventive strategies to limit the mortality caused by obesity.

Structural impairments of the heart associating with ETB mutation, a cause of Hirschsprung disease

Background HSCR, a colonic neurocristopathy affecting 1/5000 births, was suggested to associate with cardiac septal defects and conotruncal malformations. However, we question subtle cardiac changes maybe more commonly present due to multi-regulations by HSCR candidate genes, in this instance, ETB. To investigate, we compared the cardiac morphology and quantitative measurements of sl/sl rat to those of the control group.Methods Eleven neonatal rats were generated from heterozygote (ETB+/-) crossbreeding. Age and bodyweight were recorded at time of sacrifice. Diffusion-staining protocols with 1.5% iodine solution was completed prior to micro-CT scanning. All rats were scanned using an in vivo micro-CT scanner, Caliper Quantum FX, followed by two quality-control scans using a custom-built ex vivo micro-CT system. All scans were reviewed for gross cardiac dysmorphology. Micro-CT data were segmented semi-automatically post-NLM filtering for: whole-heart, LV, RV, LA, RA, and aortic arch. Measurements were taken with Drishti. Following image analysis, PCR genotyping of rats was performed: five sl/sl rats, three wildtype, and three heterozygotes. Statistical comparisons on organ volume, growth rate, and organ-volume/bodyweight ratios were made between sl/sl and the control group. Results Cardiac morphology and constituents were preserved. However, significant volumetric reductions were recorded in sl/sl rats with respect to the control: whole heart (38.70%, p-value = 0.02); LV (41.22%, p-value = 0.01), RV (46.15%, p-value = 0.02), LA (44.93%, p-value = 0.06), and RA (39.49%, p-value = 0.02). Consistent trend was observed in growth rate (~20%) and organ-volume/bodyweight ratios (~25%). On the contrary, measurements on aortic arch demonstrated no significant difference among the two groups. Discussion Despite the presence of normal morphology, significant cardiac growth retardation was detected in sl/sl rat, supporting the likely association of cardiac anomalies with HSCR, at least in ETB-/- subtype. Structural reduction was likely due to a combination of failure to thrive from enteric dysfunction, alterations to CaNCC colonization, and importantly coronary hypoperfusion from elevated ET-1/ETA-mediated hypervasoconstriction. Little correlation was detected between aortic arch development and sl/sl rat, supporting minor ETB role in large vessels. Although further clinical study is warranted, HSCR patients may likely require cardiac assessment in view of congenital cardiac defects.

A Metric Fractionator for Estimation of Total Cell Number in Paraffin-Embedded Flat or Hollow Organs

The physical fractionator is a convenient and practical solution for estimation of total cell number in a regulatory toxicology setting because it is insensitive to shrinkage allowing for paraffin processing/embedding and does not require measurement of the reference or organ volume. The principle involves sampling a known fraction of an organ in one or more steps and counting the total number of cells present in the final sample, physical disector section pairs. The total cell number in the organ is estimated by multiplying the cell count in the final fraction by the inverse of the sampling fraction(s). The key feature of the design is that tissue shrinkage due to paraffin processing occurs before the organ is uniformly sampled. Another requirement is that thermal expansion or contraction is avoided during the preparation of disector sections from the individual embedded subsamples, which ensures that the disector sections represent a known constant fraction. This vertical physical fractionator with subsampling is a simple and fast estimator to obtain precise and robust estimates of total cell number in large flat or hollow organs that do not prolong routine necropsy procedures. It is compatible with paraffin processing, avoids exhaustive sectioning, and allows for the collection of routine histopathology sections.