Understanding the Scientific Basis of Post-traumatic Stress Disorder (PTSD): Precision Behavioral Management Overrides Stigmatization.

Post-traumatic stress disorder (PTSD) is a severe polygenic disorder triggered by environmental factors. Many polymorphic genes, particularly the genetic determinants of hypodopaminergia (low dopamine function), associate with a predisposition to PTSD as well as substance use disorder. Support from the National Institutes of Health for neuroimaging research and molecular, genetic applied technologies has improved understanding of brain reward circuitry functions that have inspired the development of new innovative approaches to their early diagnosis and treatment of some PTSD symptomatology and addiction. This review presents psychosocial and genetic evidence that vulnerability or resilience to PTSD can theoretically be impacted by dopamine regulation. From a neuroscience perspective, dopamine is widely accepted as a major neurotransmitter. Questions about how to modulate dopamine clinically in order to treat and prevent PTSD and other types of reward deficiency disorders remain. Identification of genetic variations associated with the relevant genotype-phenotype relationships can be characterized using the Genetic Addiction Risk Score (GARS®) and psychosocial tools. Development of an advanced genetic panel is under study and will be based on a new array of genes linked to PTSD. However, for now, the recommendation is that enlistees for military duty be given the opportunity to voluntarily pre-test for risk of PTSD with GARS, before exposure to environmental triggers or upon return from deployment as part of PTSD management. Dopamine homeostasis may be achieved via customization of neuronutrient supplementation "Precision Behavioral Management" (PBM™) based on GARS test values and other pro-dopamine regulation interventions like exercise, mindfulness, biosensor tracking, and meditation.

Giant benign mesenchymoma of the breast.

The term mesenchymoma refers to a group of mixed tumors that are composed of two or more mesenchymal elements, excluding fibrous tissue, not ordinarily found together within the same tumor. Mesenchymomas occur most commonly in the renal and perirenal regions with rare occurrence in the breast. We describe what to our knowledge is the first report of a giant benign mesenchymoma of the breast. The clinical presentation, course, and treatment of a patient with this condition is discussed. Clinicians should be aware that benign mesenchymomas may involve the breast and simulate a malignant breast neoplasm.

Distribution of tenascin, cellular fibronectins and integrins in the normal, hyperplastic and neoplastic breast.

We present immunolocalization data on tenascin (Ten), and extradomains A and B (EDA-, EDB-) and oncofetal (Onc-) isoforms of cellular fibronectin (cFn), and alpha 1-6 and alpha v and beta 1-4 integrin subunits on cryosections of normal human breast, the spectrum of fibrocystic disease (FCD), and benign and malignant breast neoplasms. Fetal breast ducts were surrounded by broad Ten bands; adult breast ducts and acini were encompassed by thin continuous rims. In FCD, Ten was detectable and was clearly enhanced around hyperplastic ducts. Fibroadenomas showed uneven Ten periductal reactions while in all carcinomas, the stroma showed extensive and strong reactions that were most intense at the tumors' invasive edge. EDA-cFn's distribution was similar to Ten's but the staining was stronger while EDB- and Onc-cFn were virtually restricted to fetal breasts and carcinomas. In the normal adult breast, alpha 1,2,3 and alpha 6, and B1 and beta 4 integrins were detected in myoepithelial cells; weaker staining was also noted in the basolateral aspect of luminal cells; this profile was retained--and at times enhanced--in FCD, fibroadenomas and in situ carcinomas in which myoepithelial elements were present. In carcinomas, particularly in those of high grade, integrins tended to be reduced. However, mucinous carcinomas showed enhanced expression and the emergence of alpha 5 integrin that was not in the normal repertory; also a subset of infiltrating lobular carcinomas showed prominent alpha 1 and alpha 6 and beta 1 and--rarely--beta 4 staining distributed in delicate cytoplasmic processes (kinetopodia). These data indicate that the complex cell-matrix and cell-cell interactions of the normal breast are slightly altered in hyperplastic processes and benign neoplasms whereas profound chances occur in carcinomas. The latter display enhanced Ten and EDA- and Onc-cFn expression in particular, while most integrins appear decreased. Notably, mucinous and some lobular carcinomas display enhancement of certain integrins. The conspicuous localization of integrins in kinetopodia may be significant in relation to the invasive behavior of lobular carcinomas.

Experimental model of congenital syphilis.

Female LSH hamsters infected with Treponema pallidum subsp, endemicum before pregnancy or during early pregnancy transmit a form of syphilis to the fetus that is similar to human congenital syphilis. The offspring develops rhinitis, skin rash, failure to thrive, and hepatosplenomegaly. T. pallidum is detectable in their livers, spleens, and nasal secretions. Immunoglobulin M antibodies are detected in the serum.

Macrophage function in chronic experimental alcoholism. I. Modulation of surface receptors and phagocytosis.

In the present study, we have assessed peritoneal, alveolar and splenic macrophages for expression of Fc and C3b surface receptors and their ability to function in immunophagocytosis. We have also measured their oxidative burst response by the nitroblue tetrazolium dye (NBT) reduction method. Our studies revealed that macrophages harvested from chronic alcoholic rats expressed surface C3b and Fc receptors, with significantly higher surface density than macrophages of litter-mate controls (matched for sex and nutritional calories). However, the ability of macrophages from alcoholic rats to phagocytize through C3b and Fc receptors was significantly impaired. In addition, the ability of peritoneal macrophages from alcoholic animals to ingest non-opsonized Candida albicans and to reduce NBT dye was markedly compromised. Abnormalities of macrophage function may, at least in part, account for an increased susceptibility of alcoholic patients to infection.

Functional analysis of T-cell subsets in chronic experimental alcoholism.

In order to obtain a better understanding of immune system function in chronic alcoholism, we have assessed primary B-cell responses to helper T-cell independent (TI) and dependent (TD) antigens in chronic alcoholic Sprague-Dawley male rats fed totally liquid diet containing ethanol. Pair-fed littermates received the same diet except that carbohydrates isocalorically replaced ethanol, which accounted for 36% of the total calories. The ability of alcoholic animals to mount primary in vivo splenic plaque-forming cell (PFC) responses to TI pneumococcal polysaccharide type III (SIII) was elevated throughout 50 days of observation when compared to pair-fed controls; serum antibody responses to SIII paralleled the enhanced PFC responses. Primary in vivo B-cell responses to antigen sheep red blood cells (SRBC), a TD antigen, were initially elevated but were found to be significantly suppressed 30 days after chronic ethanol consumption. The degree of immunosuppression increased with length of chronic ethanol consumption. The elevated primary splenic PFC responses to TI (SIII) may be attributed to loss of T-suppressor cell control, since alcoholic rat spleen cells did not respond to low-dose priming with SIII. We suggest that either loss of function and/or actual depletion of accessory and regulatory cells (T-suppressor and T-helper) may be responsible for irregularities in B-cell function observed during chronic alcoholism. T-cell subset enumeration using fluorescein-labelled monoclonal antibodies revealed that a sequential T-helper and T-suppressor loss occurred several days following dysfunction of these T-cell subsets in splenic populations, suggesting that a combination of numerical and dysfunctional changes in lymphocyte subpopulations may be responsible for the immunological alterations observed in chronic alcoholics.

A potential vaccine for cocaine abuse prophylaxis.

Presently, there are estimated to be 1.5 to 2.0 million individuals infected with HIV-1 in the U.S. and about 12 million worldwide. In the U.S. over 90% of reported cases of AIDS occurred among two subgroups, homosexual males and intravenous substance abusers (IVSAs). Currently, there is no anti-cocaine addiction medication available. In order to explore vaccination as an alternative means for protection against cocaine addition, we immunized inbred male Fisher rats with either cocaine emulsification in complete Freund adjuvant (CFA) or with cocaine conjugated with keyhole limpet hemocyanin (KLH), as carrier plus CFA. Animals were initially immunized with 0.1 mg/animal of cocaine or cocaine-KLH. Animals were given a booster with the corresponding agents after 4 weeks. Ten animals/group were used. Controls received normal saline at the time of immunizations. These animals were injected, intraperitoneally, with 25 mg/kg of cocaine, and were examined for the analgesic effect of cocaine by the hot plate method. The average analgesic effect of cocaine was significantly reduced (p greater than 0.03) in animals immunized with cocaine-KLH (13.77 s) as compared to saline controls (21.6 s). Fifty percent of the animals (5/10) in the cocaine-KLH group and 33% of the animals (3/9) in the cocaine immunized group appeared completely resistant to the effect of cocaine on the central nervous system (CNS). We also have determined the levels of cocaine-specific antibodies produced by each animal by an ELISA method. Degree of protection from cocaine seems to correlate well with the amount of anti-cocaine antibodies produced by each animal (-0.61).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine-deaminase-associated immunodeficiency. I. Differential sensitivities of lymphocyte subpopulations exposed to 2-deoxycoformycin in vivo.

In order to obtain a better understanding of the degree of immune dysfunctions caused by the absence of adenosine deaminase, we gave a single i.p. injection of 2'-deoxycoformycin (2-dcf), a potent inhibitor of the enzyme ADA at various doses into adult Syrian hamsters. These animals were examined for their ability to mount primary in vivo antibody responses to helper T cell dependent (Th-d) and helper T cell independent (Th-ind) antigens. Hamsters treated with 0.5 mg/kg of 2-dcf mounted enhanced splenic plaque-forming cell (PFC) responses to sheep erythrocytes, a Th-d antigen, and to pneumococcal polysaccharide type III (SIII), a Th-ind antigen. Treatment of animals with 1.0 mg/kg of 2-dcf resulted in a significantly depressed (P less than 0.001) PFC response to Th-d antigen, but a further enhanced response to Th-ind antigen. One mechanism which may be responsible for such a dichotomous response to these two types of antigens was selective dysfunction of T cell subpopulations. At higher doses (1.5-4.0 mg/kg), PFC responses to both types of antigens were significantly suppressed. Immunoenhancement at low doses of 2-def was attributed to an increased susceptibility of T suppressor cells to 2-dcf. This hypothesis was confirmed by priming the 2-dcf-treated animals with low-dose Th-ind antigens. These animals failed to induce low-dose tolerance by stimulation of antigen-specific suppressor T cell subsets. At low doses, B cells and T helper cell functions were found to be intact, as further confirmed by priming the animals with the carrier keyhole limpet haemocyanin (KLH) and challenging with trinitrophenyl-KLH. This dose-dependent selective susceptibility of various T cell subpopulations and B cells may explain the heterogeneity of clinical, biochemical and immunological parameters observed in children with ADA deficiency severe combined immunodeficiency.

In vitro cloning of tumor stem cells in semi-solid media containing agar and agarose.

The formation of tumor stem cell colonies in vitro has been studied by comparing the growth of three mouse teratocarcinoma derived cell lines and one human teratocarcinoma derived cell line in semi-solid media containing either agar or agarose. We show that agaroses should be used in higher concentrations than agar to obtain comparable results. The maximum number of colonies were obtained in agarose over a broader range of concentrations (1%-4% for SeaPrep and 0.5%-2% for SeaPlaque agarose) than in agar, which allowed anchorage-independent growth of tumor cells only over a narrow concentration range (0.3%-0.5%). Overall, the preparation of media containing agarose was less cumbersome than preparation of agar-containing media, primarily because agaroses gelled more slowly and remained liquid in the physiological temperature range. Furthermore, the transfer of colonies from semi-solid media containing agarose to solid surface tissue culture dishes was much more efficient than the transfer of colonies from agar. The stock solutions of SeaPrep agarose could be kept ready for use for extended periods of time. All these features show that the low melting point agarose has considerable advantages over agar for preparation of semi-solid media for anchorage-independent tumor cell growth.

Comparison of image analysis of imprints with flow cytometry for DNA analysis of solid tumors.

Quantitative analysis of cellular DNA content may be clinically useful for several solid tumors. The technology for this analysis by flow cytometry or image analysis has existed for several years but has not been widely used, except in a handful of specialized research institutions. Recently, however, relatively inexpensive image analyzers intended for use by hospital pathologists have been introduced that can analyze DNA content from cytology or imprint specimens which are readily obtainable from solid tumors. We report here an assessment of this technology for analysis of tumor imprint specimens, using flow cytometry of tissue blocks as the standard for comparison. We used image analysis equipment on Feulgan-stained imprint preparations from 31 tumors and compared the histograms with those obtained by flow cytometric analysis of archival tissue blocks from the same tumors. The ploidy descriptors (diploid, tetraploid, and aneuploid) were concordant for the two methods in 27 specimens, with three specimens yielding discordant results and one specimen considered unevaluable by image analysis. The image analysis method using imprints appeared to have several advantages over flow cytometry, including lower instrument cost, no need to dissociate paraffin blocks or fresh tissue, and ability to analyze very small samples. Somewhat lower resolution of the histograms, extremely localized tissue sampling, and possibly greater risk of occasionally obtaining unevaluable preparations were disadvantages. Microcomputer-based image analysis performed on imprints appeared to be a viable alternative to flow cytometric analysis of tissue blocks for quantitative DNA analysis of tumor specimens.

Differential distribution of tenascin in the normal, hyperplastic, and neoplastic breast.

We studied by immunohistochemistry the distribution of tenascin with the monoclonal antibody 100EB2, and compared it with that of laminin in breast tissue samples from fetal, adult resting, lactating, and aging parenchyma, variants of fibrocystic disease, fibroadenomas, cystosarcoma phylloides, and ductal and lobular carcinomas. Monoclonal antibodies were applied to cryosections by the avidin-biotin-complex method; selected samples were studied by double immunofluorescence, and by Western blot analysis. In adult resting and aging breasts, tenascin immunoreactivity was noted in the periductal and periacinar stromal regions as thin irregular bands; in the lactating breast, broader periductal bands were observed. In these samples, laminin immunoreactivity was a single continuous line around ducts, acini, and vessels. In fetal breasts, tenascin appeared as thick periductal bands, whereas laminin remained as a delicate single line. In FCD, tenascin increased around ducts showing hyperplasia, papillomas and apocrine metaplasia, whereas laminin retained its delicate linear pattern. Similar patterns were seen in fibroadenomas and cystosarcoma phylloides with variable tenascin reactivity in the stroma beyond the ducts. Tenascin immunoreactivity was markedly increased around ducts containing in situ carcinoma appearing as broad bands, whereas that of laminin showed a linear, frequently discontinuous appearance. Prominent stromal tenascin immunoreactivity was seen in infiltrating ductal and lobular carcinomas, whereas laminin was virtually absent save for scattered lines. The abundance of tenascin in the carcinomatous stroma contrasted with its scarcity in the non-neoplastic stromal regions. By Western blotting, both chains of tenascin with molecular weights 250,000 and 180,000 were shown in ductal and lobular carcinomas as well as in normal breast. Tenascin immunoreactivity was noted in the periepithelial stromal regions of adult resting and aging breast ducts and acini. The amount of tenascin was moderately enhanced in certain physiologic conditions (fetal growth, gestation), as well as hyperplasias, dysplasias (fibrocystic disease) and benign tumors, whereas it was markedly enhanced in intraductal and infiltrating carcinomas. During fetal mammary development, adult physiologic and pathologic hyperplasias, and in carcinomas, the increasing tenascin reactivity contrasted with the stable or decreasing laminin reactivity.