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Targeted deletion of Insm2 in mice result in reduced insulin secretion and glucose intolerance
Lin Wang1,2,
Zhong Sheng Sun1,2,
Bingwu Xiang1,
Chi-ju Wei3,
Yan Wang2,
Kevin Sun4,
Guanjie Chen4,
Michael S. Lan5,
Gilberto N. Carmona6,
Abner L. Notkins6 &
Tao Cai ORCID: orcid.org/0000-0002-6447-76551,6
Neurogenin3 (Ngn3) and neurogenic differentiation 1 (NeuroD1), two crucial transcriptional factors involved in human diabetes (OMIM: 601724) and islet development, have been previously found to directly target to the E-boxes of the insulinoma-associated 2 (Insm2) gene promoter, thereby activating the expression of Insm2 in insulin-secretion cells. However, little is known about the function of Insm2 in pancreatic islets and glucose metabolisms.
Homozygous Insm2−/− mice were generated by using the CRISPR-Cas9 method. Glucose-stimulated insulin secretion and islet morphology were analyzed by ELISA and immunostainings. Expression levels of Insm2-associated molecules were measured using quantitative RT-PCR and Western blots.
Fasting blood glucose levels of Insm2−/− mice were higher than wild-type counterparts. Insm2−/− mice also showed reduction in glucose tolerance and insulin/C-peptide levels when compared to the wild-type mice. RT-PCR and Western blot analysis revealed that expression of Insm1 was significantly increased in Insm2−/− mice, suggesting a compensatory response of the homolog gene Insm1. Similarly, transcriptional levels of Ngn3 and NeuroD1 were also increased in Insm2−/− mice. Moreover, Insm2−/− female mice showed a significantly decreased reproductive capacity.
Our findings suggest that Insm2 is important in glucose-stimulated insulin secretion and is involved in the development pathway of neuroendocrine tissues which are regulated by the transcription factors Ngn3, NeuroD1 and Insm1.
Pancreatic endocrine cells play a key role in glucose metabolism by regulating the synthesis and secretions of islet hormones, such as insulin and glucagon. Mature islets contain five different types of endocrine cells. Ngn3, a proendocrine marker, is essential for islet cell development [1]. Biallelic mutations in Ngn3 have been identified to cause permanent neonatal diabetes [2, 3]. NeuroD1, which is directly activated by Ngn3, participates in the maintenance and differentiation of mature islet cells [4]. Heterozygous mutations in NeuroD1 have been identified to be associated with maturity-onset diabetes (OMIM: 601724) and type 2 diabetes mellitus [5, 6]. Although much efforts have been made to decipher the mechanism of differentiation and maintenance of pancreatic endocrine cells, the molecular basis of how Ngn3 and NeuroD1 function in the islets remains under investigation.
Previously we reported the isolation of two homologous genes, INSM1 (a.k.a. IA-1) [7] and INSM2 (INSM transcriptional repressor 2; a.k.a. IA-6) [8], from human pancreatic islet cells. Two additional genes, named PTPRN and PTPRN2 (a.k.a., IA-2 and IA-2beta), were also isolated from human islet cells, which have been turned out to be major autoantigens in patients with type 1 diabetes and are involved in the secretion of hormones and neurotransmitters [9, 10]. The encoded C2H2 zinc-finger proteins by INSM1 and INSM2 belong to the SNAIL/GFI1/INSM transcriptional repressor superfamily (i.e., SNAG-domain mediated transcription factors) that plays an important role in the developmental processes and molecular pathogenesis of various human conditions [11, 12]. For instance, mutations in GFI1 and GFI1B have been identified in patients with neutropenia (OMIM: 600871) [13] and affected members of a family with platelet-type bleeding disorder-17 (OMIM: 604383) [14, 15], respectively.
Functional studies demonstrated both INSM1 and INSM2 are direct targets of Ngn3 and NeuroD1 [8, 16]. During the embryonic development, the Insm1−/− knockout mice showed a significantly reduced insulin production, suggesting a key role for Insm1 in islet β-cell differentiation and maturation [17, 18]. Recent studies demonstrated that Insm1 cooperates with Neurod1 and Foxa2 to maintain mature pancreatic beta-cell function [19]. Insm2 also was found to be expressed in developing endocrine cells peaking from E11.5 to E13.5 and activated in Ngn3/NeuroD1-transduced pancreatic epithelial duct cells [8]. Numerous clinical studies showed that Insm1 is a sensitive and highly specific marker for various tumors, such as neuroendocrine differentiation in primary lung neoplasms [20], Merkel cell carcinoma [21], small cell carcinoma of the prostate [22], head and neck tumors [23] as well as insulinoma.
To explore further the biological and pathophysiological role of Insm2 in pancreatic islets, we developed an Insm2−/− mouse using the CRSPR-Cas9 technique, and measured blood glucose, insulin and C-peptide levels of the Insm2−/− mice [9]. Furthermore, we measured transcriptional levels of the transcription factors Ngn3, NeuroD1 as well as the homolog gene Insm1 in Insm2−/− mice.
Generation of Insm2-deficient mice
The pST1374-NLS-flag-linker-Cas9 (#44758) and pUC57-sgR (#51132) plasmids were commercially obtained (http://www.addgene.org/). Single guide RNA (sgRNA) was designed to target the coding region of the mouse Insm2 gene (GenBank accession number: NM_020287.2). Mouse embryos were injected with Cas9 and sgRNA and then transferred into pseudo-pregnant mice to give birth to chimeric mice. Homozygous Insm2−/− global knockout mice were obtained by breeding of Insm2+/− mice and wild-type C57BL/6J. All experimental procedures were carried out in accordance with protocols approved by the NIDCR Animal Care and Use Committee (#12-641) and the Animal Usage Committee of Wenzhou Medical University (#31571301). Statistical analysis was performed using the Student's t test for unpaired comparisons.
Genotyping and quantitative real-time PCR
Primers for genotyping (forward: 5′-gtctcagctataaagcgggc-3′, reverse: 5′-aattggaacggatacaggga-3′) were located on flanking sides of the deleted region of Insm2. PCR products were genotyped by Sanger sequencing. Total RNAs were isolated from mouse brain and pancreas tissues using TRIZOL reagents (Life Technology, Rockville, MD, USA). The concentration of RNAs was determined by NANO 2000 (Thermal Scientific, Amarillo, TX, USA). Real-time PCR was performed using Mx3000p System with a two-step cycling program (Stratagene): 95 for 10 min, 40 cycles of 95 °C for 1 min and 60 °C for 30 s. Primers included for following genes: Insm2 (forward: 5′-gctccggcagctcctacc-3′; reverse: 5′-ggctcctccggtgaggatt-3′), Insm1 (5′-ggagtacgctgacccgttcg-3′; 5′-aagaccttggcgcactctgg-3′), Ngn3 (5′-aagagcgagttggcatgagcaag-3′; 5′-gcgttggtccgctatgcgcag-3′), NeuroD1 (5′-cttggccaagaactacatctgg-3′; 5′-ggagtagggatgcaccgggaa-3′), mouse Insulin 1 gene (5′-ccttagtgaccagctataatcagag-3′; 5′-cacttgtgggtcctccactt-3′) and Insulin 2 gene (5′-tcagcaagcaggaagcctatcttcc-3′; 5′- cacttgtgggtcctccactt-3′) [24]. Gapdh PCR product was used as a loading control.
Western blots and antibodies
Proteins were isolated from mouse brain tissues for Western blots with following antibodies: anti-INSM2 rabbit antibody (1:800) [8], INSM1 rabbit antibody (1:1000, ab170876, Abcam, Shanghai, China), and anti-GAPDH antibody (1:1000, Santa Cruz (6C5), USA). The bands were visualized using enhanced chemiluminescence detection reagents (Applygen Technologies, Beijing, China) and detected by the FluorChem E imaging system (Cell Biosciences, Santa Clara, USA).
Histological staining and immunostaining
Genotyped mice aged 10–23 weeks were sacrificed for isolation of pancreatic tissues, which were then treated in 10% formalin buffer for paraffin embedding. 5 μm thick sections on slides were stained with hematoxylin and eosin (H&E) for further analysis. Monoclonal anti-insulin antibody (1:250, Clone K36AC10, Sigma, USA) and anti-INSM2 rabbit antibody (1: 100) were also tested for immunostainings of mouse pancreas tissues [8].
Intraperitoneal glucose tolerance test
Targeted mice aged 10–23 weeks were fasted for 14 h, followed by glucose injection (2 g/kg body weight via intraperitoneal injection). Venous blood was drawn from the tail vein at 0 (just before the injection), 15, 30, 60, 90 and 120 min after the injection of glucose. Blood glucose level was measured using a portable glucometer (Bayer, Elkhart, IN) as described previously [25].
Insulin secretion
Glucose (3 g/kg body weight) was injected intraperitoneally into male and female mice aged 10–23 weeks. Insulin levels were measured using rat Insulin Ultrasensitive ELISA Kit and mouse C-peptide ELISA Kit (ALPCO, Salem, NH, USA). Blood samples were collected from the mouse orbital vein and centrifuged at the speed of 3000 rpm for 10 min at 4 °C and stored at − 80 °C until use.
Characterization of Insm2 −/− mice
The offsprings of the CRISP-Cas9 mice were genotyped by PCR and Sanger sequencing, which showed a 25-bp deletion in coding region of the sole exon of the Insm2 gene (Fig. 1a). This deletion resulted in a frameshift mutation p.S36fsX67 (FWGPPASHLRRTTPSGARVAATAPAPAPRGRRAPSCAARSWSAACA LRSLRSPSPAPPP SAPRRPRP)*, thereby disrupting the reading frame of Insm2. Wild-type (Insm2+/+), heterozygous (Insm2+/−), and homozygous (Insm2−/−) mice were identified by tail DNA PCR and Sanger sequencing with the allele-specific primers (Fig. 1b).
Genotyping and characterization of Insm2−/− mice. a Targeted deletion of 25-bp (c.108_132delAGTCCCCGACCGGGCTCCTCCGGTG) in the coding sequence region (CDS) of Insm2 (GenBank acc. no., NM_020287.2). b Sanger chromatograms show the target sequence regions in wild-type (WT), heterozygous (HET), and homozygous (HO) Insm2−/− mice. c Quantitative RT-PCR analysis indicates that Insm2 mRNA is markedly decreased in the Insm2−/− mouse brain as compared to normal controls. d Western blotting shows no detectable INSM2 protein in Insm2−/− mouse brain tissues. e, f H&E staining reveals no significant structure alterations in Insm2−/− mouse pancreatic islets
Quantitative RT-PCR analysis of mRNAs extracted from the brain tissues of Insm2−/− mice showed that the mRNA levels of Insm2 were dramatically decreased, probably due to unstable Insm2 mRNAs with the frameshifted deletion, compared to the wild type mice (Fig. 1c). The mRNA level of Insm2 in pancreas tissue was also significantly decreased as compared to the wild type mice (not shown). Western blot analysis, using rabbit anti-INSM2 sera to measure protein expression level in mouse brain extracts, clearly showed no visible expression of INSM2 protein in the homozygous Insm2−/− as compared to Insm2+/+ mice (Fig. 1d).
Phenotype analysis
Physical examination of the Insm2−/− mice revealed no gross development abnormalities. Body weight of Insm2−/− male or female mice at 8, 16, and 24 weeks of age did not differ from the Insm2+/+ mice. However, fertility of Insm2−/− female mice was significantly reduced. Specifically, after mating with Insm2−/− male mice, of the 16 mating pairs of Insm2−/− female mice, only five (31.25%) gave birth but with a smaller litter size (4 vs. 9 in normal mice, P < 0.0001). In a sharp contrast, all Insm2−/− male mice had normal reproductive capacity when mating with Insm2+/– female mice. However, the underlying pathogenesis of the significantly reduced fertility observed here needs further investigation.
Pancreatic tissue analysis using H&E staining in more than 30 slides did not show any significant alterations in terms of the number and size of islets in Insm2−/− mice (Fig. 1e, f). However, fasting blood glucose levels of the Insm2−/− male mice at 16 weeks of age (Fig. 2a) and female mice at 16- and 24 weeks of ages were elevated (Fig. 2b) as compared to the Insm2+/+ mice.
Analysis of glucose tolerance, insulin secretion, and Insm2 homologous or regulatory genes. a, b Fasting glucose levels are elevated at 24 weeks Insm2−/− male and at 16 and 24 weeks old Insm2−/− female mice. Values are presented in mean ± SEM from eight animals per group in three separate experiments (**P < 0.01; ***P < 0.001). c, d Glucose tolerance tests in male and female Insm2+/+ and Insm2−/− mice. After overnight fasting, d-glucose (2 g/kg body weight) was injected intraperitoneally, and blood glucose levels were measured at different times as indicated. Values are presented in mean ± SEM from 14 mice per group in three separate experiments (**P < 0.01; ***P < 0.001). e, f Blood insulin and C-peptide levels in response to intraperitoneal glucose in male and female Insm2−/− mice aged 16–24 weeks were measured and compared to Insm2+/+ mice at 15 min. Blood samples were drawn from the tail vein using heparinized capillary tubes before and after glucose injection
Glucose tolerance tests also showed significantly elevated glucose levels in both male and female Insm2−/− mice (Fig. 2c, d). ELISA analysis showed that serum insulin levels in these mice were significantly lower than that in the Insm2+/+ mice (i.e., 0.56 ng/ml vs 1.37 ng/ml in male mice and 0.57 ng/ml vs 1.31 ng/ml in female mice, respectively, Fig. 2e). Similarly, basal blood C-peptide levels in the Insm2−/− mice also were significantly decreased compared to the wild type mice (0.14 ng/ml vs 0.31 ng/ml in males and 0.11 ng/ml vs 0.23 ng/ml in female, respectively, Fig. 2f).
In fact, quantitative RT-PCR using primers of either the mouse insulin 1 gene (Ins1) or insulin 2 gene (Ins2) showed that their transcriptional levels in pancreatic tissues of Insm2−/− mice were only 60–65% of the wild-type mice (Fig. 3a, b). Immunostainings using anti-mouse insulin antibody also showed reduced insulin expression in Insm2−/− mouse islet beta cells compared to the wild-type islet cells (Fig. 3c).
Insulin expression levels in pancreatic islet cells. a, b Expression of the Ins1 and Ins2 gene is decreased in the pancreas tissues of the Insm2−/− mice compared to the wildtype controls. c Immunostainings show lower expression of insulin in the islet cells of Insm2−/− mice compared to the wildtype controls
Expression levels of Insm2-associated regulators
Since Insm2-encoded protein shares significant homology as well as similar protein structure with Insm1 [8], this suggested that the expression of Insm1 might be increased as a compensatory response to the deficiency of Insm2. RT-PCR analysis of the Insm2−/− brain tissue mRNAs showed a 2.5-fold increase of Insm1 expression as compared to that of wild type mice (Fig. 4a). Western blotting with the anti-INSM1 antibody showed that the INSM1 protein levels in Insm2−/− mice were 2 to 3-folds higher than that of wild type mice (Fig. 4b). Islet tissue was not examined because there is no detectable expression of Insm1/Ngn3/NeuroD1 in adult mouse islet cells.
Expression levels of Insm1, Ngn3, and NeuroD1. a, b Insm1 mRNA and its encoded protein levels extracted from brain tissues and measured by quantitative RT-PCR and Western blots. c, d Transcriptional levels of Ngn3 and NeuroD1 extracted from brains and analyzed by qRT-PCR. The results represent the average of three independent experiments in triplicates. Data are means ± SEM. *P < 0.01; **P < 0.001; ***P < 0.0001
Given that the transcription factors Ngn3 and NeuroD1 were previously found to promote Insm2 expression via binding to the proximal E-boxes of the Insm2 promoter [8], we examined their transcriptional response to the deletion of Insm2. RT-PCR analysis showed that expression levels of Ngn3 and NeuroD1 mRNAs were significantly increased in the Insm2−/− mice compared to that of Insm2+/+ mice (Fig. 4c, d). Taken together, our findings suggest that the Ngn3 and NeuroD1 will elicit the expression of Insm2 in a number of neuroendocrine tissues.
The human INSM2 gene was mapped to chromosome 14q13.2 and showed a broader spatial-temporal expression pattern in neuroendocrine, heart, and liver tissues [8], whereas the INSM1 expression was restricted in neuroendocrine tissues and tumors [26, 27]. INSM2 belongs to the SNAI1 (OMIM: 604238)/GFI1 (OMIM: 600871)/INSM1 (OMIM: 600010) family of transcriptional repressors (OMIM: 614027). Intriguingly, the mouse Insm2 gene was found to be methylated and silenced in liver tumors of SV40 T antigen transgenic mice [28]. In contrast to the earlier death of Insm1−/− mice with disrupted development of insulin-producing beta cells [17], the phenotypes we observed in Insm2−/− mice were relatively subtle. Although no obvious diabetic symptoms were observed in Insm2−/− mice, elevated fasting glucose, elevated glucose levels in glucose tolerance test, lower serum insulin secretion, and lower C-peptide levels are indicators that deletion of Insm2 affects pancreatic islet cell functions (Fig. 5). Understanding of the regulatory mechanisms of critical transcriptional factors, such as Ngn3, NeuroD1, and Insm1, in islet development is important for making insulin-producing cells for prospective stem cell therapy to treat diabetes [29,30,31].
Interactions of several crucial transcriptional factors in pancreatic islet insulin-producing cells. Arrows in black indicate direct or regulatory interactions; arrows in red point to the phenotypic alterations due to deleterious mutations previously identified in affected individuals or specific knockout of the corresponding gene in mice
In depth analyses of Insm1, Ngn3, and NeuroD1 in Insm2−/− mice revealed that their increased expressions could be a compensatory response to the deficiency of Insm2 because both Insm1 and Insm2 share a common activation pathway through Ngn3 and NeuroD1 in islet cells [8]. Previous study also demonstrated that endogenous Insm2 expression was activated in Ngn3/NeuroD1-transduced pancreatic epithelial duct cells [8]. Therefore, the phenotype severity in pancreatic islets of Insm2−/− mice could be lessened through the increased Insm1 expression. In fact, similar to the expression pattern of Insm2 in brain [8], Ngn3 and NeuroD1 are also abundantly expressed in brain tissues, especially in developing and adult hippocampus [32, 33]. Therefore, the increased expression levels of Ngn3 and NeuroD1 in brain tissues should be a compensatory response to the deletion of Insm2 in mice. As previously reported, Ngn3 and NeuroD1 positively regulate Insm2 expression through the binding of the E-box of the Insm2 promoter [8]. It would be interesting to search additional insulin-pathway related phenotypes in the double knockout mice harboring homozygous Insm2−/− and heterozygous Insm1+/− alleles.
Furthermore, genetic analysis by GWAS in a cohort of Africa Americans with type 2 diabetes revealed a significant association of the disease to a SNP (rs1952392, MAF = 0.0188; P < 0.001) at the proximal promoter region of the human INSM2 gene (personal communication) [34]. A previous genetic study also showed an association of microsatellite polymorphisms at 14q13.2 with type 2 diabetes mellitus in Latvian and Finnish populations [35]. Interestingly, INSM2 was recently found to upregulate the expression of the ultraconserved (uc) RNA uc.372, which in turn suppresses the maturation of miR-195/miR-4668 to regulate expression of genes related to lipid synthesis and uptake in liver [19]. Given the risk of non-alcoholic fatty liver disease in type 2 diabetes mellitus (see recent meta-analysis) [36, 37], it is worth noting whether the deletion of Insm2 in mice affects the hepatic lipid accumulation and thus attributes to the risk of type 2 diabetes.
Based on the important role of the INSM2 gene in neuroendocrine tissues as well as the preliminary clinical findings, potential mutations and altered expressions of the INSM2 gene in neuroendocrine-associated disorders, such as diabetes and infertility, can be screened utilizing next-generation sequencing technologies. In a cancer-related study, Insm2 was found to be methylated and silenced in liver tumors of SV40 T antigen transgenic mice [28], suggesting its role in tumor inhibition. Among 64 different mammalian cell lines, intriguingly, INSM2 appears to be exclusively expressed in SH-SY5Y cells (a neuroblastoma cell line, Human Protein Atlas, https://www.proteinatlas.org). Further studies are needed to learn whether or not INSM2 plays a role in tumorigenesis and functions as a prognostic marker in neuroblastoma.
In conclusion, we demonstrated that deletion of Insm2 affects glucose-stimulated insulin secretion and glucose tolerance. We also showed that Insm2 is involved in the Ngn3/NeuroD1 pathway in neuroendocrine tissues. Whether Insm2−/− mice induce islet cell differentiation and proliferation defects and/or develop type 2 diabetes, further experiments under more challenging conditions, such as high-fat and high-sugar diet [38, 39], are required to test this hypothesis.
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TC, ZSS, and ALN conceived the project. LW, YW and KS performed the experiments. LW, BX, CW, GC, GNC, ZSS and TC contributed to the acquisition, analysis and interpretation of the data. TC wrote the paper. All authors were involved in drafting the article or revising it and gave their approval for the final manuscript to be published. TC is the guarantor of this work and takes full responsibility for the content of the manuscript. All authors read and approved the final manuscript.
We thank the NIDCR Gene Targeting Facility for technique assistance.
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
This investigation was carried out according to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996). All animal protocols were approved by the NIDCR Animal Care and Use Committee (#12-641) and the Animal Usage Committee of Wenzhou Medical University (#31571301).
This study was supported by Zhejiang Province Nature Science Research funding (LY13C070004) and Science and Technology Supporting Program, Ministry of Science and Technology of China (M2014BAI02B01 to ZSS). This research was supported in part by the Intramural Research Program of the NIDCR, NIH.
Institute of Genomic Medicine, Wenzhou Medical University, Wenzhou, China
Lin Wang
, Zhong Sheng Sun
, Bingwu Xiang
& Tao Cai
Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, China
& Yan Wang
Multidisciplinary Research Center, Shantou University, Shantou, Guangdong, China
Chi-ju Wei
Center for Research on Genomics and Global Health, NHGRI, NIH, Bethesda, MD, USA
Kevin Sun
& Guanjie Chen
Department of Pediatrics, Louisiana State University Health Sciences Center, New Orleans, LA, USA
Michael S. Lan
Experimental Medicine Section, NIDCR, NIH, B30/Rm112, 30 Convent Dr., Bethesda, MD, 20892, USA
Gilberto N. Carmona
, Abner L. Notkins
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Wang, L., Sun, Z.S., Xiang, B. et al. Targeted deletion of Insm2 in mice result in reduced insulin secretion and glucose intolerance. J Transl Med 16, 297 (2018) doi:10.1186/s12967-018-1665-6
Accepted: 14 October 2018
Human diabetes
Knockout mouse
Pancreatic islets
Combination strategies

redpajama_set_name: RedPajamaCommonCrawl


Volunteer in Nepal after the earthquake, take your trip off the beaten path with a one-month cultural exchange and volunteering experience.
The Exchange for Change program is a unique volunteer program that enables participants to integrate into community activities for a month while familiarizing themselves with Nepalese culture.
During the program, the volunteers get to live with Nepali families, experience Nepalese culture and become part of the local community. The volunteers are integrated into the organization's work in the village in the fields of agriculture, the youth movement, education in schools and in the women's groups.
The program is designed for travelers, professionals, and anyone interested in contributing their time to experience a unique cultural experience.
Tevel b'Tzedek is an organization that works and plans with the local communities, and therefore the work in the village is done over several years; the volunteers are part of this ongoing and sustainable process.
The program's duration is about a month, and includes 3-5 days of orientation in Kathmandu, where volunteers participate in language lessons, familiarity with Nepalese culture, and deepening their knowledge of globalization, developing countries, community work, and the Jewish world.
At the end of the orientation, the volunteers will move to the village. At this stage, the volunteers will be divided into families' homes and will integrate into various volunteer activities together with the local team: empowerment of women, work with youth, agriculture, and education.
Towards the end of the month, the volunteers will return to a seminar in Kathmandu to document their work in order to transfer their knowledge and experience to the next group, thus creating a continuum of work in the village.
The cost of the program is $500, a sum that covers the expenses of the host families in the villages, food, travel and the study seminar. The cost does not cover flights, medical insurance and visas.
The program takes place during the the tourist seasons in Nepal (September-November and February-June).
We are unable to run the program during the summer months (June-August) due to the weather conditions during the monsoon season.
The program operates in the District of Kavre in an area called Dhunkharka, which is about a five-hour drive from Kathmandu, the capital of Nepal. The work is carried out in two different villages of about 200 families. Most of the families are from the ethnic Tamang group. The group of volunteers will be divided between the two villages, which are a 30-minute walk from each other.
In order to register for the program, please fill in the registration form!
After that, a representative from the organization will contact you and set up a personal interview according to the program date you wish to register for.


Lamotrigine is an anticonvulsant used in human medicine, but in companion animals it can cause seizures and severe cardiac arrhythmias.
Dextromethorphan is a common ingredient in over-the-counter cold, congestion, and flu medications, and pets who ingest it may need veterinary care.
Albuterol, an asthma treatment, is a common toxic ingestion. The Animal Poison Control Center explains the best treatments for pets who've been exposed to it.
Why dogs find certain things tasty is still beyond us, but sunscreen is one of those items the ASPCA Animal Poison Control Center gets regular calls about in the summer. Bone up on sunscreen ingredients so you're ready!
How would you treat a dog who ingested atomoxetine? This case study from the ASPCA Animal Poison Control Center explains.


Sunglasses by the brand ROCCO BY RODENSTOCK Unisex Sunglasses, with reference S0004725, with a(n) Formal, Modern and Retro/Vintage style.Rocco 312 D 54 19 they have a(n) Full Rim frame made of Acetate / plastic with a(n) Square shape in Blue colour. Their size is 54mm and their bridge width is 19mm, with a total frame width of 73mm. They have Organic, Blue, Gradient lenses. These glasses are suitable for prescription lenses.
These are top of the range glasses from the brand ROCCO BY RODENSTOCK, made of high-quality materials, modern glasses that will give you a look that matches your image. Buy Rocco 312 D 54 19 sunglasses in an easy and safe way at the best price in our online store.


This is a pellet fired, Wi-Fi enabled portable grill that operates on 12vdc and has great reviews! I, being impatient as always, decided to do some searching around Pheonix and came upon Grills Galore in Scottsdale which both had the unit in stock, and honored the current sale price! That night, we had grilled filet mignon and it was DELICIOUS!
The Davy Crockett can hold a temperature from 150 F to 550 F, and thus can be used for slow smoking all the way up to full on searing and grilling! The Wi-Fi app lets you not only monitor the temperature of your grill, but also that of your food with it's included food probe. It also lets you adjust the grill temp and even set up custom profiles!
For example, if you wanted to heat the grill to 250 F, and keep it there until your food reached an internal temperature of 165 F, then turn the grill down to 200 F for 2 hours, then down to 150 F until you turn it off, all that can be configured into a profile that you can save, and re-use later! It will also send you alerts when each step is reached and can do so (and be controlled) remotely via their cloud connection. A true Geek's Grill! It also gets better with time. They continue to release firmware upgrades for the grill to improve temperature accuracy, integrate the cloud option, etc.
I've now cooked several filet mignon steaks on the grill, burgers, and brats. I've not ventured into smoking yet but hope to soon.


text:
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All posts tagged "campus recreation facility"
Home / Posts tagged "campus recreation facility"
By Brittany Howard on December 2, 2019
Utah State University Logan, UT MISSION STATEMENT The Department of Campus Recreation provides wellness and healthy lifestyle-oriented programs, events and services to Utah State University...
The University of Nevada, Las Vegas
By Brittany Howard on November 4, 2019
The University of Nevada, Las Vegas Las Vegas, NV MISSION STATEMENT The University of Nevada, Las Vegas (UNLV) campus recreational services is a premier campus...
Red Rocks Community College
By Brittany Howard on October 7, 2019
Red Rocks Community College Lakewood, CO MISSION STATEMENT Our mission is to inspire holistic wellness by engaging the Red Rocks community in developmental and educational...
How a Bike Shop can Benefit Your Rec Center
By Brittany Howard on August 13, 2019
In 2007, Kennesaw State University (KSU) first opened the bike shop as a service center for their recreation facility rental bikes. One year later, that...
Why Your Campus Rec Center Needs an Escape Room
By Brittany Howard on August 6, 2019
In spring of 2018, Florida State University (FSU) launched their first escape room as an additional offering to their FSU Challenge team building program. Since...
Stonehill College Easton, MA MISSION STATEMENT The Athletic Department of Stonehill College endeavors to provide the students, staff, faculty, alumni and community constituencies a bedrock...
Should Your Facility Invest in Indoor Turf?
By Brittany Howard on July 30, 2019
At the University of Vermont, the multipurpose tennis and turf facility provides space for a variety of campus recreation programs throughout the year. Because...
Thinking Outside the Wellness Box at Clemson University
By Brittany Howard on July 9, 2019
What started as a mini convenient store in the Fike Recreation Center, featuring Swann Fitness Center, at Clemson University has now transformed into the Wellness...
The Facility Enhancement and Renovation Process
By Eric Maki on July 2, 2019
As a follow-up to last week's story on renovation don'ts, Eric Maki, the senior director of campus recreation at West Chester University, shares his advice...
Georgetown University Washington, D.C. MISSION STATEMENT Georgetown University, as a leading American and Jesuit institution, is committed to the cultivation of the whole person;...
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The residents of the small town of Wadebridge, Cornwall, are aiming to become the first solar- and renewable-energy powered town in the UK. It has an ambitious short-term goal of generating 30% of its electricity from renewables by 2015, which is about a third of the town's electricity supply. The town's struggle is being shown on Youtube as a four part mini-series.
With the UK government pledging 50% emissions cut by 2025, new solar initiatives are popping up all through the country.The Wadebridge campaign to convert to renewable energy is being led by the Wadebridge Renewable Energy Network (WREN), – a not-for-profit co-operative. The plan is to put thousands of panels on the roofs of local homes, schools and businesses, allowing them to use the clean electricity, and collect the feed-in tariff for their community fund.
WREN is starting installations with 100 homes, half of which will be buying the systems themselves, and half of which will be funded through a partnership between solar provider Solarcentury and Triodos Bank But the controversial proposals by the British government to limit the Feed-In-Tariff (payment for generating electricity) to small-scale projects are in danger of altering the town's plans.
The first episode was uploaded to YouTube on May 20 and outlines the aims of the project and the difficulties involved in winning over some of Wadebridge's residents. The next episode wll be made avaiulable in two week.
Watch the first episode: http://www.youtube.com/watch?v=kZA43jMAlE4.


There shall not be any use of forced labor, whether in the form of prison labor, indentured labor, bonded labor or otherwise of whatsoever means.
We always discourage child labor and no person shall be employed at an age of younger than 15 years or where the Governing law allows such employment. Or younger than the age for completing compulsory education if the age in the country of manufacturing is greater than 15.
We always treat our workers with respect and dignity. Workers shall not be subject to any physical, verbal, sexual or psychological harassment or abuse in connection with their employment.
Workers should be employed and compensated based upon their ability to perform their job, rather than on the basis of gender, race, religious or cultural beliefs.
Employers/contractors shall pay workers for all work completed and shall pay at least the minimum wage required by law or the prevailing industry wage, whichever is higher and shall provide legally mandated benefits.
Employers/contractors should provide to workers, for each pay period, a clear and understandable wage statement that includes days worked, wage or piece rate earned per, hours of overtime, bounces, allowances and a description of any and all deductions.
We prefer using time cards manual or electronic to be used for hourly workers. For "piecework" worker an industry recognized system, capable of audit verification, should be used.
As part of normal business operation, workers shall be entitled to at one full day off out of each seven-day week. In the case of those countries where laws do not limit the hours of work, workers should not be required to work on a regular basis in excess of a regular work week plus 12 hours overtime. Workers shall not be asked or required to take work home or off premises.
It is understood that overtime is often required in the manufacturing process. In addition to compensation for regular hours of work, the worker shall be compensated for overtime hours at such a premium rate as legally required or in countries where there is no legal standard, at industry standards. In no event shall this be at a rate less than the regular hourly rate. Factories shall carry out operations in such ways that limit overtime to a level that ensures productive and humane working conditions.
All standards set forth in this code of conduct are subject to compliance with applicable local law. All factories shall operate in full compliance with the laws of their respective country of manufacture. If any standard set forth in this code of conduct is, in the factories judgement, deemed to violate an applicable local law it must advise ApparelnBags.com promptly in writing.
Factories should have adequate circulation and ventilation.
Free portable water should be reasonably accessible to all workers throughout the working day.
Workers should have access to reasonably clean and private toilet facilities in adequate number for each gender.
There is to be no use of corporal punishment.
Work areas should be sufficiently lit, so that manufacturing tasks may be safely performed.
All equipment should be tested regularly and properly maintained. Operational safety devices should be installed on equipment where appropriate.
Where appropriate, factories should make available personal protective equipment to workers performing hazardous tasks.
Hazardous and combustible materials should be stored in a secured and well-ventilated areas.
Each factory should have at least one well-stocked first aid kit.
We require that each enclosed manufacturing facility have an adequate number of clearly marked, unlocked exits in order to provide for an effective evacuation in case of emergency. By adequate, it is meant that each workstation should be within 200 feet of an exit.
Aisles, exits and stairwells should be kept clear of any obstructions that could prevent the orderly evacuation of workers.
Each facility should have appropriate fire prevention capability based upon the size of the facility and number of worker normally occupying the premises.
Fire extinguishers should be properly maintained and inspected annually, tagged with the most recent inspection date, and properly deployed throughout the premises to be visible and accessible to workers in case of fire.
Factories should conduct a minimum of one emergency evacuation drill per year.
We created the Productive Partnership Program (PPP) so that our employees could take part in activities that express the principles and values of ApparelnBags.com social responsibility objectives. The program gives employees a chance to share their skills and experiences and help those in need foster their personal, social and professional development.
Primarily we endeavor utmost to curtail our production needs by local resources. However, besides this, we through our subsidiary provide manufacturing services from Pakistan and Bangladesh. These subsidiaries are in apparel business since decades.
Here ApparelnBags.com becomes the team leader and provide employment to hundreds of skill and non-skilled laborers in the field of apparel and clothing.
Here our experienced staff conducts a special training program and share their skills and experience for the needy people.
We conduct this mentoring twice a year and the successful candidates get promotions and a special increase in the salary or the wages are allocated.
ApparelnBags.com holds its factories accountable that the merchandise produced for it is manufactured in compliance with this code of conduct.
ApparelnBags.com expects its factories to monitor their compliance to this code of conduct.
ApparelnBags.com will be permitted to check employees files, including time cards and wages statements.
We always provide a copy of supplier code of conduct, duly translated into the native languages of the workforce and should be prominently displayed with each facility where ApparelnBags.com Inc merchandise is manufactured.
ApparelnBags.com Inc under any circumstances cannot tolerate any vendor that directly or indirectly, through its subcontractors, violates the laws of the country where the merchandise is manufactured.


Using this mod will give URLs posted in your topics an automatic title. I saw this on another forum I visit and thought it to be pretty nifty, so I coded this up for SMF.
If the page or title cannot be found, the URL in the post is not changed.
This change is made when the post is made or edited, it is not done on the fly, as that may cause lags in load time.
There are also options in the admin interface for Posts so you can turn this feature on or off, and also limit the number of URLs to convert.
convert_urls.php is a tool that can be used to convert all urls in existing messages to titled urls. I have tested this on a message table with 12,000 messages and it worked as it should, but I cannot promise that it will always work. Any errors should be reported to the topic for this mod.
1. Download from the SimpleMachines.org Mod Site.
2. Upload to your forum using the built-in Package Manager.
3. Install using the built-in Package Manager.
1.1a: Bug with URLs that had regex characters in them.
1.1: No code changes, just updated installer for 1.1.3.
1.1: Added settings to 'Posts And Topics'.  Removed XSS vulnerability.


text:
Nicole Beland was the Men's Health "Girl Next Door" until the May 2009 issue. Beland's column from Men's Health has been converted into a book called Ask the Men's Health Girl Next Door (). She attended Union College in Schenectady, NY.

She also contributed to its sister publication, Women's Health, as a Deputy Editor, until she left to work at Cosmopolitan as Executive Editor. Along with Ted Spiker, Beland contributed to an advice column on the Women's Health website. A former senior editor at Cosmopolitan and Mademoiselle magazines, she currently lives in Carroll Gardens, Brooklyn.

She is also the author of several books including The Cocktail Jungle: A Girl's Field Guide to Shaking and Stirring (2003), Girl Seeks Bliss (2005), and Sex: The Whole Picture (2005). She has appeared as a health and relationships expert on CNN Headline News, The Early Show, Good Morning America, The Today Show, and The Best Damn Sports Show Period.

Articles for Women's Health

 "A Girl Walks into a Bar", Fall 2004
 "Are You on the Right Pill?", February/March 2005
 "The Pursuit of Happiness", April/May 2005

References

External links
 Nicole Beland on Amazon.com
 Nicole's advice column on Women's Health
 Nicole Beland profile on Gothamist
 "Ask the Men's Health Girl Next Door" featured in USA Today

Year of birth missing (living people)
Living people
Union College (New York) alumni
People from Carroll Gardens, Brooklyn
American women columnists
20th-century American journalists
20th-century American women writers
21st-century American journalists
21st-century American women writers
Writers from Brooklyn

redpajama_set_name: RedPajamaWikipedia


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We air this series for free typically once a year, sometimes twice, as our way to give back and help as many people as possible.
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Episode 1: The Sacred Plant…Examined
In this exciting season premiere, you'll discover what makes the sacred plant such a powerful healing agent and learn about the dark history of its prohibition. You'll also hear heart-wrenching yet inspirational stories from multiple patients including a 4 time Grammy Award winner, how the Hyde family snuck the sacred plant into the hospital for their 2 year old son, Cash…and how he beat cancer twice and many more.
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Episode 2: Stopping Pain, Insomnia, Anxiety, and Autoimmune Conditions
Discover how the sacred plant addresses multiple symptoms at once and helps patients get off of addictive and harmful pharmaceutical drugs.
Episode 3: Ending Suffering For Our Children: Autism, Seizures, and More
Experience for yourself how these autistic and epileptic children have made miraculous improvements using the sacred plant…when years of taking prescription medication and traditional meds have failed them.
Episode 4: Treating Cancer Part 2
In a continuation of where we left off in Episode 4 of Season 1, we'll take you deep we inside the cancer-sacred plant connection with patient and doctor interviews. See for yourself how the sacred plant heals, treats, and even beats this dreaded disease.
Episode 5: Accessing The Sacred Plant: The Unjust Laws and How to Navigate Them
We'll show you how the sacred plant is grown and processed for medical use and what you can do if you live in a state where the sacred plant isn't yet legal. You'll also meet patients and parents who fought for legal access and won.
Episode 6: Extending the Quality and Quantity of Life For Us and Our Beloved Pets
Discover how the sacred plant is used for wellness and prevention in seniors…and for treating your pet(s).
Episode 7: Treating PTSD + Selecting Your Medicine, Dosage, and a Qualified Doctor
Experience how the sacred plant has helped veterans and trauma victims with PTSD and take a behind-the-scenes look into how to buy medicine at a dispensary, select the right dosing and find a qualified sacred plant doctor.
Starts December 12th
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We'll show you how the sacred plant is grown and processed for medical use and what you can do if you live in a state where sacred plant isn't yet legal. You'll also meet patients and parents who fought for legal access and won.
Experience how the sacred plant has helped veterans and trauma victims with PTSD and take a behind-the-scenes look into how to buy medicine at a dispensary, select the right dosing and find a qualified the sacred plant doctor.
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You'll hear the research, stories, and case studies that didn't make it into the regular series episodes because of time constraints.
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In these 2 videos, John Malanca answers your most pressing questions about the myths and methods on using The Sacred Plant safely and effectively to prevent, treat, and even beat most major diseases.
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Short on time? Skim through these transcripts to pull out the "gold nuggets" from every episode and interview.
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INSTANT access to 3 How-To Videos
In the first how-to video Dr. Patricia Frye explains how to make a homemade THCA; in video two Myriam's Hope co-founders, Diana & Jeff, give tips on how-to about making your own medicine and what materials to use; and in the third video Dr. Eric Zielinski provides his simple formula on how to make your own homemade oil.
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Value - $37
Dr. Allan Frankel is a UCLA-trained doctor who has practiced medicine in the Los Angeles area since 1979. And in this interview, he shares facts you need to know about using the sacred plant as medicine, including, which medical conditions respond well to the sacred plant, how to use the sacred plant without getting high, and more. (You'll receive a video and transcript of the interview.)
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Have you ever wondered if hemp CBD is effective as sacred plant CBD? Or what dosages and ratios you should use for treating conditions such as cancer, epilepsy, autism, parkinson's and more? Great news, John Malanca along with one of the world's leading authorities on the sacred plant, Dr. Allan Frankel provide insights to these questions and many more in this interactive Q&A session. (You'll receive a video and transcript of the interview.)
BONUS #3: Expert Interview with Dr. Dustin Sulak
Millions of Americans endure the misery and hopelessness of depression, anxiety, and insomnia. The good news is, the sacred plant can help manage these conditions. In this interview, Dr. Dustin Sulak discusses how the sacred plant can soothe these symptoms, proper dosing, and more. (You'll receive a video and transcript of the interview.)
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BONUS #5: Q&A with Dr. David Bearman
Do you have questions about using the sacred plant to treat pain? In this 2-Part Q&A video, you'll hear from Dr. David Bearman, a doctor with over 40 years of medical experience, on how to use the sacred plant to treat pain. Dr. Bearman has worked with more than 3,000 patients. You'll get insights, expert advice, and action steps you can take to finally loosen pain's grip on your life.
BONUS #6: Patient Interview with Cancer Patient, Don Adams
For most of us, there's nothing scarier than the thought of getting cancer. This year alone, more than 1.7 million new cases of cancer will be diagnosed in the United States. When Don Adams received his cancer diagnosis six years ago he thought it was the beginning of the end. But in this video you'll see how the sacred plant beat Don's cancer -- and saved his life.
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When it comes to treating cancer, the sacred plant is not a one-size-fits-all solution. In this video, Don Adams talks about the dosage and strains he used to beat cancer. Together with your host and advocate, John Malanca, they answer questions from cancer patients. If you want to know how to use the sacred plant to beat cancer, you'll get the facts and details you need here.
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In this interview, Sebastien Cotte tells the story of his son Jager. Sadly, Jager developed a rare terminal disease when he was an infant. But Jager was able to live a longer and better life thanks to the sacred plant. This interview is essential viewing for anyone suffering from a terminal disease.
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If you're a parent or grandparent, you want the best for the children in your life. So what to do when a child you love suffers anxiety, ADHD, or sickness and disease? In this Q&A, Sebastien Cotte, and your host, John Malanca, answer questions about using the sacred plant to help our kids live happier, healthier lives.
BONUS #10: Patient Interview with Cancer Patient and Mom Of Cancer Patient, Shannan Ajluni
In this interview you'll meet Shannan Ajluni, a mother, patient, advocate, and breast cancer survivor. For years, Shannan relied on pharmaceuticals and prescription meds to fight her cancer. But they didn't stop her pain and suffering. So she decided to try the sacred plant. And you'll get to see and hear how that decision changed her life.
BONUS #11: Interactive Q&A with Cancer Patient and Mom Of Cancer Patient, Shannan Ajluni
Shannan Ajluni beat breast cancer. But that's just the start of her story of courage and survival. You see, Shannan's two sons -- Oliver and Joey -- also face harrowing health challenges: Oliver has a rare terminal cancer and Joey was diagnosed with autism and Tourette's syndrome. In this Q&A session Shannan reveals how the sacred plant has helped her entire family, bringing hope where they once were hopeless.
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And you'll get instant access to all of them when you order The Sacred Plant: Healing Secrets Examined today.
Why Purchase The Sacred Plant: Healing Secrets Examined
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You can watch each episode of The Sacred Plant: Healing Secrets Examined for free online starting Wednesday, December 12th. So why should you buy it?
Here are 5 reasons why:
Reason #1: You Get Instant and Unlimited Access
Right now, you can only watch each episode of The Sacred Plant: Healing Secrets Examined once, when it airs. But what if work, family, or your busy schedule keeps you from seeing an episode? Or, what if you want to rewatch an episode to make sure you don't miss out on any of its health-transforming information? Sadly, you wouldn't be able to.
But when you choose to own The Sacred Plant: Healing Secrets Examined you get INSTANT and UNLIMITED access to all 7 episodes. You can watch them any time, day or night. You can watch them again and again to make sure you don't miss out on anything. And you can rest easy knowing that whenever you need access to this powerful information, it will be right at your fingertips.
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Do you know a friend, loved one, or maybe even a neighbor whose health and life would improve if they knew the truth about ? Now's your chance to help.
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But you'll need to act fast. Because to ensure rush delivery we're only making 1,000 units of The Sacred Plant: Healing Secrets Examined available. And to keep things fair, they're available on a first come, first served basis. So if you want to avoid waiting on your copy, I encourage you to order now.
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Order The Sacred Plant: Healing Secrets Examined today and you can begin watching it and all the bonuses today! Also, if you ordered the physical package, you'll be holding it in your hands in a few days.
So by purchasing you can immediately discover how to use the sacred plant to find freedom from chronic pain, manage autoimmune conditions, prevent and even reverse diseases like cancer, and more.
But to get access to this health-transforming information, you'll have to act fast.
Reason #5: Your Order Helps Save Lives
We believe the sacred plant has the potential to transform millions of lives for the better. Unfortunately, it's still far more difficult than it should be for good people to access this amazing sacred plant.
That's why we're donating a portion of proceeds to three organizations that are focused on providing the sacred plant to people who can't afford it, offering one-on-one medical consulting, and helping to change the law so more people can access it.
The three charities we currently support are:
Our goal is to donate at least $100,000 to these extraordinary charities through The Sacred Plant: Healing Secrets Examined before the end of the year.
Your order today brings us closer to our goal. It also helps us achieve our mission to educate and empower people to be healthy, happy, and pain-free using the healing powers of the sacred plant.
In fact, we re-invest 100% of our profit to film and produce more content, market it to a larger audience of people who need to see it, manage daily operations, and more.
So it's important you know that when you support our mission, you're supporting an organization that was built on the foundation of caring and giving.
What's more, when you order today you risk nothing, thanks to our…
180 Day Money-Back Guarantee
You can order now with complete peace-of-mind… Because we're giving you 180 days to try owning The Sacred Plant: Healing Secrets Examined.
We're certain you're going to love it. But if for some reason you're not satisfied, you can get a full refund withing 180 days
No questions asked, and we'll still be friends.
PLUS, you can keep all your bonuses just for giving it a try!
But you'll need to hurry…
We only have 1,000 physical copies of The Sacred Plant: Healing Secrets Examined in stock. So, the first 1,000 Gold or Diamond orders will receive guaranteed rush holiday delivery.
So to ensure you get your copy (as well as all your instant bonus gifts) please use the order form below now.
(Please note: If we're already sold out of The Sacred Plant: Healing Secrets Examined when you try to order, your name will be added to a waiting list. Thank you for your understanding.)
Get Instant and Unlimited Access to ALl Episodes, Extended Interviews, Plus Trancripts and Over $760 in Bonus Gifts!
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"WOW! Season 2 was amazing and heartbreaking/heartwarming on so many levels! Thank you for your tenacity and transparency and the HOPE and HELP that is The Sacred Plant!"
– L. Lake
"I can't thank you enough for all I've been learning, and I so appreciate all you and your team do to bring truth and help to those so desperately in need."
– M. Choufani
"Thank You for your hard work and dedication to getting the truth out to the world. Programs like this can and do open the minds of people, and hopefully the change we want is just around the corner for all people in this world.
Let's all work together to make the health revolution that is desperately needed happen."
– M. McLean
"Sincere thanks & very grateful for all that you're doing for us, sharing with us and for this 2nd series of The Sacred Plant." -D. Corrigan
"Everyone should take time to watch and learn about this Sacred Plant. It will help so many people!"
-M. Koh
You can order now with complete peace-of-mind... Because we're giving you 180 days (that's six calendar months!) to try The Sacred Plant: Healing Secrets Examined in the comfort of your own home.
If you're not 100% satisfied, you can get a full refund within 180 days of your purchase -- and still get to keep the digital bonuses as our gift to you.
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About The Sacred Plant: Healing Secrets Examined
How can I join the movement and support the mission?
Can you see how the sacred plant has such and impact on countless lives?
You can play a special part in this important movement by supporting our mission at the link below and giving life, health, and hope to your loved ones, those who are suffering, and also to yourself.
When you get Unlimited Access to The Sacred Plant: Healing Secrets Examined, you not only get a copy of entire series PLUS some amazing bonuses…
You also get a SECOND copy to give to a loved one, AND here's the great news…right now, you can get up to 60% off at the link below when you order TODAY. This discount is for a LIMITED time only, so jump on this now to save.
100% of the profits are pumped back into research, education, charitable giving, and getting treatment for extreme-needs cases.
You are saving lives.
When you support the movement, you support those who are suffering.
Learn more above.
Which packages qualify for the "Buy One, Get One Free" buddy pass?
Our desire is for you to be able to share The Sacred Plant: Healing Secrets Examined with friends or family—or even your doctor! So all packages have a Buy One, Give One Free buddy pass:
If you order the digital-only Silver package, we'll send you a coupon for a friend to create their own username/password in our membership portal with all the downloads.
If you order the Gold package, we'll send you an extra set of DVDs.
If you order the Diamond combo package, we'll send you BOTH the coupon and the extra set of DVDs!
When will I receive my DVD Package in the mail?
It typically takes 1-2 weeks to receive your physical package… IF you order now.
After this presale period, we'll place one order at the end of the series to get the best price so we can reinvest more money into the mission.
We will ship the orders, first come, first send. Meaning that the earlier you purchase your physical package, the earlier you will receive it.
Please keep in mind that we're not BIG PHARMA with endless streams of income, and we're only able to produce what has been ordered, so please be patient with us as we produce, print, and ship your orders as quickly as possible.
We also know that you're eager to get started, and this is why you will have INSTANT access to all of the videos, and all transcripts, other interviews, and bonuses, as soon as you choose unlimited access.
Choose your package above.
How do I get email notifications and links to the Episodes as they air?
Register here to get all updates and live links to each Episode as they air: https://two.thesacredplant.com/docuseries
I didn't get my confirmation email. What do I do?
If for whatever reason you did not get your registration confirmation, here are some steps you can take to ensure all is in order:
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How many episodes is Season 2: Healing Secrets Examined?
Just like our first series, Season 2, Healing Secrets Examined is a 7-part episode docuseries. When you register you will receive an email. If you don't receive this email within an hour, please check your SPAM folder, and if it's not there, please contact us via our support desk here.
Please contact The Sacred Plant Customer Support with regard to your concern/inquiry by submitting a ticket here.
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text:
Alastor facilis is a species of wasp in the family Vespidae.

References

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