Gene expression and characterization of clonally derived murine embryonic brown and brite adipocytes.

White adipocytes store energy, while brown and brite adipocytes release heat via nonshivering thermogenesis. In this study, we characterized two murine embryonic clonal preadipocyte lines, EB5 and EB7, each displaying unique gene marker expression profiles. EB5 cells differentiate into brown adipocytes, whereas EB7 cells into brite (also known as beige) adipocytes. To draw a comprehensive comparison, we contrasted the gene expression patterns, adipogenic capacity, as well as carbohydrate and lipid metabolism of these cells to that of F442A, a well-known white preadipocyte and adipocyte model. We found that commitment to differentiation in both EB5 and EB7 cells can be induced by 3-Isobutyl-1-methylxanthine/dexamethasone (Mix/Dex) and staurosporine/dexamethasone (St/Dex) treatments. Additionally, the administration of rosiglitazone significantly enhances the brown and brite adipocyte phenotypes. Our data also reveal the involvement of a series of genes in the transcriptional cascade guiding adipogenesis, pinpointing GSK3ß as a critical regulator for both EB5 and EB7 adipogenesis. In a developmental context, we observe that, akin to brown fat progenitors, brite fat progenitors make their appearance in murine development by 11-12 days of gestation or potentially earlier. This result contributes to our understanding of adipocyte lineage specification during embryonic development. In conclusion, EB5 and EB7 cell lines are valuable for research into adipocyte biology, providing insights into the differentiation and development of brown and beige adipocytes. Furthermore, they could be useful for the characterization of drugs targeting energy balance for the treatment of obesity and metabolic diseases.

Frequency Specific Optogenetic Stimulation of the Locus Coeruleus Induces Task-Relevant Plasticity in the Motor Cortex.

The locus ceruleus (LC) is the primary source of neocortical noradrenaline, which is known to be involved in diverse brain functions including sensory perception, attention, and learning. Previous studies have shown that LC stimulation paired with sensory experience can induce task-dependent plasticity in the sensory neocortex and in the hippocampus. However, it remains unknown whether LC activation similarly impacts neural representations in the agranular motor cortical regions that are responsible for movement planning and production. In this study, we test whether optogenetic stimulation of the LC paired with motor performance is sufficient to induce task-relevant plasticity in the somatotopic cortical motor map. Male and female TH-Cre + rats were trained on a skilled reaching lever-pressing task emphasizing the use of the proximal forelimb musculature, and a viral approach was used to selectively express ChR2 in noradrenergic LC neurons. Once animals reached criterial behavioral performance, they received five training sessions in which correct task performance was paired with optogenetic stimulation of the LC delivered at 3, 10, or 30 Hz. After the last stimulation session, motor cortical mapping was performed using intracortical microstimulation. Our results show that lever pressing paired with LC stimulation at 10 Hz, but not at 3 or 30 Hz, drove the expansion of the motor map representation of the task-relevant proximal FL musculature. These findings demonstrate that phasic, training-paired activation of the LC is sufficient to induce experience-dependent plasticity in the agranular motor cortex and that this LC-driven plasticity is highly dependent on the temporal dynamics of LC activation.

FAM129B is a cooperative protein that regulates adipogenesis.

FAM129B is one of Niban-like proteins described in neoplastic cells and implicated in melanoma cell invasion, but no reports have been published on FAM129B and cell differentiation. We show that FAM129B is early and transiently expressed and crucial for 3T3-F442A adipogenesis. Fam129b is expressed downstream of the early genes Cebpb, Klf4, Klf5 and Srebf1a, but upstream of Pparg2 since knockdown of Fam129b blocked Pparg2 expression and adipose differentiation. Glycogen synthase kinase 3 beta activity, a crucial kinase for adipogenesis, and the ERK1/2 are involved in FAM129B phosphorylation as part of the adipogenic program. Phosphorylated FAM129B is crucial for Pparg2 expression and the lipogenic gene expression downstream of Pparg2, and hence for adipogenesis. Fam129b knockdown reduced adipocyte cluster formation and size, regulating commitment and clonal amplification. In vivo, BAT, inguinal and epidydimal fat expressed Fam129b, suggesting a role in adipose tissue development. We conclude that FAM129B is a cooperative protein that regulates differentiation during the early stages of adipogenesis.

Louise Eisenhardt (1891-1967): World Renowned Neuropathologist, Brain Tumor Registry Director, First Editor-In-Chief of Journal of Neurosurgery, and President of Harvey Cushing Society.

BACKGROUND: This historical account reviews the course and lasting impact of Dr. Louise Eisenhardt (1891-1967) in neurosurgery. METHOD: The writing of this project was sparked by the discovery of original scientific and bibliographical information about Eisenhardt, testimony on personal relationships, and viewpoints after comprehensive compilation of information. It is a thorough review of literature on Eisenhardt and reflects the scope and depth of these prior works. RESULTS: It begins with the decisive influence of Harvey Cushing's mentorship and academic; follows Eisenhardt's impact on the development of modern neuropathology; discusses the Eisenhardt-Percival-Bailey-Cushing collaboration on gross brain specimens and histological classification of brain tumors; recounts Cushing's creation of a neuropathologist team for the Brain Tumor Registry working asynchronously with the Pathology Department at Peter Bent Brigham Hospital; Eisenhardt's aid in the development of intraoperative analysis of brain tumors; her career as a neuropathologist; her contributions as Editor-in-Chief of the Journal of Neurosurgery; and her preservation of the Brain Tumor Registry at Yale University School of Medicine estimated the largest and most valuable databank of information in the history of medicine. Eisenhardt served as President, Historian, and Secretary-Treasurer of the Harvey Cushing Society, the professional organization now known as the American Association of Neurological Surgeons, and was senior lecturer for members of the Congress of Neurological Surgeons, constituents of NEUROSURGERY Publications. CONCLUSIONS: Our article provides glimpses into the personality of Dr. Louise Eisenhardt and her marked impact on neurosurgery and allied neurosciences.

Pipeline for fluorescent imaging and volumetric analysis of neurons in cleared mouse spinal cords.

Precisely measuring the number and somatic volume of neurons in the central nervous system at single-cell resolution is technically challenging. Here, we combine multiple techniques to address this challenge in optically cleared mouse spinal cords. We describe in vivo neuron labeling approaches, tissue-clearing technology, light sheet fluorescence microscopy, and machine learning-guided imaging analysis. This combination provides a precise determination of the cell number and somatic volume of any neuron population in the spinal cords.

Schenck's knee dislocation (KD) I injury: An uncommon pattern.

Schenck's knee dislocation (KD) I is a very uncommon pattern of KD, where the knee presents with clinical or radiographic evidence of a KD with one of the ligaments of the central pivot preserved. The correlation between imaging and physical examination is of the highest importance to correctly classify this injury. Recently, there have been reports in the literature with large numbers of Schenk's KD I, however this is due to classifying multiligament knee injuries as if they all were KDs rather to an increase of the pattern. In this report the case of an 18-year old patient that sustained a KD with a preserved posterior cruciate ligament is presented.

Abordaje anterolateral extendido y fijación con placa rim en fragmentos posterolaterales en fracturas de platillo tibial / Extended anterolateral approach and rim plate fixation for posterolateral fragments in tibial plateau fractures

Los fragmentos posterolaterales (FPLs) en fracturas de platillo tibial son frecuentes, pero difíciles de reducir y estabilizar. Actualmente existe controversia sobre cómo deben ser abordados; si bien un abordaje anterolateral es más seguro, este puede ser insuficiente para lograr una correcta reducción. Los abordajes posterolaterales y posteriores ofrecen una visión directa y permiten una fijación con ventaja biomecánica; sin embargo, son más demandantes y exponen al paciente a una mayor morbilidad. A continuación, se presenta una nota técnica sobre la reducción y fijación de FPLs con una placa rim por medio de un abordaje anterolateral extendido a través del espacio paraligamento colateral lateral.

Posterolateral fragments (PLFs) are commonly seen in tibial plateau fractures, but their reduction and fixation are challenging. There is no consensus about the ideal approach to fix this particular fragment. Even though an anterolateral approach is a safe option, it may impair a correct reduction. The posterolateral and posterior approaches offer direct visualization of the fragment, and enable a fixation with a biomechanical advantage; however, they are more demanding and expose the patient to a higher risk if morbidity. The following technical note describes the reduction and fixation of PLFs with a rim plate through an extended anterolateral approach using the paralateral collateral ligament space.

Genetically Engineering the Nervous System with CRISPR-Cas.

The multitude of neuronal subtypes and extensive interconnectivity of the mammalian brain presents a substantial challenge to those seeking to decipher its functions. While the molecular mechanisms of several neuronal functions remain poorly characterized, advances in next-generation sequencing (NGS) and gene-editing technology have begun to close this gap. The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (CRISPR-Cas) system has emerged as a powerful genetic tool capable of manipulating the genome of essentially any organism and cell type. This technology has advanced our understanding of complex neurologic diseases by enabling the rapid generation of novel, disease-relevant in vitro and transgenic animal models. In this review, we discuss recent developments in the rapidly accelerating field of CRISPR-mediated genome engineering. We begin with an overview of the canonical function of the CRISPR platform, followed by a functional review of its many adaptations, with an emphasis on its applications for genetic interrogation of the normal and diseased nervous system. Additionally, we discuss limitations of the CRISPR editing system and suggest how future modifications to existing platforms may advance our understanding of the brain.

A cellular perspective of adipogenesis transcriptional regulation.

Adipose cells store lipids in the cytoplasm and signal systemically through secretion of adipokines and other molecules that regulate body energy metabolism. Differentiation of fat cells and its regulation has been the focus of extensive research since the early 1970s. In this review, we had attempted to examine the research bearing on the control of adipose cell differentiation, some of it dating back to the early days when Howard Green and his group described the preadipocyte cell lines 3T3-L1 and 3T3-F442A during 1974-1975. We also concentrated our attention on research published during the last few years, emphasizing data described on transcription factors that regulate adipose differentiation, outside of those that were reported earlier as part of the canonical adipogenic transcriptional cascade, which has been the subject of ample reviews by several groups of researchers. We focused on the studies carried out with the two preadipocyte cell culture models, the 3T3-L1 and 3T3-F442A cells that have provided essential data on adipose biology.

Utility of venous blood gases in severe sepsis and septic shock.

Acid-base status is frequently assessed in severe sepsis and septic shock. Venous blood gas sampling is proposed as a less-invasive modality but lacks evidence within this population. The objective of this study was to evaluate the correlation and agreement between arterial blood gas (ABG), peripheral venous blood gas (pVBG), and central venous blood gas (cVBG) in severe sepsis and septic shock. We conducted a prospective, observational cohort study in subjects admitted to the medical intensive care unit. Simultaneous blood gas samples, including ABG, pVBG, and cVBG, were analyzed for correlation and agreement. Severity of illness scores revealed a mean (±SD) Sequential Organ Failure Assessment score of 7.9 ± 3.3, Simplified Acute Physiology II score of 49.3 ± 16.5, and a mortality rate of 11.9% in the intensive care unit and 16.4% in the hospital. We found a strong intraclass correlation (>0.85) for pH, partial pressure of carbon dioxide (pCO2), bicarbonate, and base excess for ABG/pVBG, ABG/cVBG, and pVBG/cVBG comparisons. Agreement by the Bland-Altman method was found for pH (bias ± SD, 0.03 ± 0.04, 0.03 ± 0.02, and 0.00 ± 0.03) but not for pCO2, partial pressure of oxygen, bicarbonate, base excess, and oxyhemoglobin saturation. In conclusion, adequate correlation and agreement between ABG/pVBG, ABG/cVBG, and pVBG/cVBG comparisons was found only for pH. The current level of evidence does not support the use of venous blood gas sampling in this setting.

Critically ill patients and gut motility: Are we addressing it?

Gastrointestinal (GI) dysmotility is a common problem in the critically ill population. It can be a reflection and an early sign of patient deterioration or it can be an independent cause of morbidity and mortality. GI dysmotility can be divided for clinical purposes on upper GI dysmotility and lower GI dysmotility. Upper GI dysmotility manifests by nausea, feeding intolerance and vomiting; its implications include aspiration into the airway of abdominal contents and underfeeding. Several strategies to prevent and treat this condition can be tried and they include prokinetics and post-pyloric feeds. It is important to note that upper GI dysmotility should be treated only when there are clinical signs of intolerance (nausea, vomiting) and not based on measurement of gastric residual volumes. Lower GI dysmotility manifests throughout the spectrum of ileus and diarrhea. Ileus can present in the small bowel and the large bowel as well. In both scenarios the initial treatment is correction of electrolyte abnormalities, avoiding drugs that can decrease motility and patient mobilization. When this fails, in the case of small bowel ileus, lactulose and polyethylene glycol solutions can be useful. In the case of colonic pseudo obstruction, neostigmine, endoscopic decompression and cecostomy can be tried when the situation reaches the risk of rupture. Diarrhea is also a common manifestation of GI dysmotility and the most important step is to differentiate between infectious sources and non-infectious sources.

Stressful Life Events and Self-Reported Postpartum Depressive Symptoms 13-24 Months After Live Birth Among Non-Hispanic American Indian/Alaska Native Mothers in Oregon: Results from a Population-Based Survey.

OBJECTIVES: We explored the association between stressful life events and postpartum depressive symptoms among non-Hispanic American Indian and Alaska Native (AI/AN) mothers. METHODS: We analyzed self-reports of stressful life events and depressive symptoms from 298 AI/AN respondents and conducted logistic regression to examine their association. RESULTS: Of the AI/AN mothers who responded, 29.7% reported depressive symptoms during their second postpartum year. Partner-related and traumatic stressful life events were significantly associated with increased risk of postpartum depressive symptoms. CONCLUSIONS: AI/AN women should receive intensive screening for depression through the second postpartum year. Programs that address stressful life events may be part of a plan to decrease postpartum depression.

Hypoventilation in neuromuscular disease.

Hypoventilation in neuromuscular disease is attributed to both respiratory muscle weakness and reduced chemoreceptor sensitivity essential in ventilatory drive. Acute or chronic respiratory failure is seen in a spectrum of neuromuscular disease; whereas some are treatable others are progressive and devastating. Sleep is associated with a reduction in ventilation and hence worsening hypoventilation. Problems with sleep may be an early indicator of further muscle weakness and should prompt the clinician for further investigation, though usefulness of pulmonary function testing, arterial blood gas analysis, and other measures may not be universally predictive. The timing of respiratory failure is variable, but knowledge of the clinical aspects, pathogenesis, and treatment of respiratory failure and hypoventilation may be helpful in evaluating the patient with neuromuscular disease. For those with progressive and terminal disease, additional factors such as end of life care, especially ventilation and cough, may be useful for the patient, caregivers, and treating medical personnel.

Marketing infant formula through hospitals: the impact of commercial hospital discharge packs on breastfeeding.

OBJECTIVES: Commercial hospital discharge packs are commonly given to new mothers at the time of newborn hospital discharge. We evaluated the relationship between exclusive breastfeeding and the receipt of commercial hospital discharge packs in a population-based sample of Oregon women who initiated breastfeeding before newborn hospital discharge. METHODS: We analyzed data from the 2000 and 2001 Oregon Pregnancy Risk Assessment Monitoring System (PRAMS), a population-based survey of postpartum women (n=3895; unweighted response rate=71.6%). RESULTS: Among women who had initiated breastfeeding, 66.8% reported having received commercial hospital discharge packs. We found that women who received these packs were more likely to exclusively breastfeed for fewer than 10 weeks than were women who had not received the packs (multivariate adjusted odds ratio=1.39; 95% confidence interval=1.05, 1.84). CONCLUSIONS: Commercial hospital discharge packs are one of several factors that influence breastfeeding duration and exclusivity. The distribution of these packs to new mothers at hospitals is part of a longstanding marketing campaign by infant formula manufacturers and implies hospital and staff endorsement of infant formula. Commercial hospital discharge pack distribution should be reconsidered in light of its negative impact on exclusive breastfeeding.

Unintended childbearing and knowledge of emergency contraception in a population-based survey of postpartum women.

OBJECTIVES: We examined the relationship between unintended childbearing and knowledge of emergency contraception. METHODS: The Oregon Pregnancy Risk Assessment Monitoring System (PRAMS) is a population-based survey of postpartum women. We analyzed data from the 2001 PRAMS survey using logistic regression to assess the relationship between unintended childbearing and emergency contraception while controlling for maternal characteristics such as age, race/ethnicity, education, marital status, family income, and insurance coverage before pregnancy. RESULTS: In 2001, 1,795 women completed the PRAMS survey (78.1% weighted response proportion). Of the women who completed the survey, 38.2% reported that their birth was unintended and 25.3% reported that they did not know about emergency contraception before pregnancy. Unintended childbearing was associated with a lack of knowledge of emergency contraception (OR 1.43, 95% CI 1.00, 2.05) after controlling for marital status and age. CONCLUSIONS: Women in Oregon who were not aware of emergency contraception before pregnancy were more likely to have had an unintended birth when their marital status and age were taken into account. Unintended birth was more likely among women who were young, unmarried, lower income, and uninsured. Given that emergency contraception is now available over-the-counter in the US to women who are 18 years of age or older, age- and culturally-appropriate public health messages should be developed to expand women's awareness of, dispel myths around, and encourage appropriate use of emergency contraception as a tool to help prevent unintended pregnancy and birth.

Breastfeeding duration and perinatal cigarette smoking in a population-based cohort.

OBJECTIVES: We examined the association between breastfeeding duration and maternal smoking before, during, and after pregnancy. METHODS: Data from the 2000-2001 Oregon Pregnancy Risk Assessment Monitoring System were used. Early weaning was defined as not breastfeeding at 10 weeks postpartum. RESULTS: At 10 weeks after pregnancy, 25.7% of mothers who initiated breastfeeding no longer breastfed. After controlling for confounders, quitters (mothers who quit smoking during pregnancy and maintained quit status after pregnancy) and postpartum relapsers (mothers who quit smoking during pregnancy and resumed smoking after delivery) did not have significantly higher risk for early weaning than nonsmokers. However, persistent smokers (mothers who smoked before, during, and after pregnancy) were 2.18 times more likely not to breastfeed at 10 weeks (95% confidence interval=1.52, 2.97). Women who smoked 10 or more cigarettes per day postpartum (i.e., heavy postpartum relapsers and heavy persistent smokers) were 2.3-2.4 times more likely to wean their infants before 10 weeks than were nonsmokers. CONCLUSIONS: Maternal smoking is associated with early weaning. Stopping smoking during pregnancy and decreasing the number of cigarettes smoked postpartum may increase breastfeeding duration.

Pregnancy intendedness and the use of periconceptional folic acid.

OBJECTIVE: Periconceptional use of folic acid can prevent birth defects, including at least 50% of neural tube defects. This study used an ongoing surveillance system to explore the association between pregnancy intendedness and women taking periconceptional folic acid. METHODS: Oregon Pregnancy Risk Assessment Monitoring System (PRAMS) surveys a stratified random sample of women after a live birth. In 1998-1999, 1867 women completed the survey (64.0% response rate); responses were weighted for nonresponse. Women were asked whether they took folic acid most days in the month before becoming pregnant. RESULTS: Overall, 33.2% of women took folic acid most days in the month before becoming pregnant, and 39.9% said that their pregnancy was unintended. Adolescent mothers were less likely to take periconceptional folic acid (9.2%) and more likely to report unintended pregnancy (62.0%) than older women. Overall, women who said that their pregnancy was intended were more likely to report that they had taken periconceptional folic acid (odds ratio: 4.75; 95% confidence interval: 3.16-7.14); after controlling for maternal age and income the odds ratio was 3.70 (95% confidence interval: 2.38-5.56). CONCLUSIONS: Women whose pregnancies were intended were more likely to have been taking periconceptional folic acid than women whose pregnancies were unintended. The importance of fertile women's taking daily multivitamins that contain 400 microg (0.4 mg) of folic acid should be stressed among women who are not contemplating pregnancy, especially adolescents and low-income women.