• BMB Reports
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• v.40 no.3
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• pp.426-431
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• 2007

To investigate the possible mechanisms of high-altitude native animals in adapting to high altitude, we cloned hemoglobin alpha-chain (alpha-chain Hb) gene from Pantholops hodgsonii, an animal species that indigenously lives at elevations of 3700-5500 m on the Qinghai-Tibetan plateau. Using reverse transcription polymerase chain reaction (RT-PCR) technique, the alpha-chain Hb gene was amplified from total RNA in the liver of the Pantholops hodgsonii. TA cloning technique was used and the PCR product was cloned into pGEM-T vector. The DNA sequence of the gene was highly homologous with sheep (99.1%), goat (98.6%), cattle (95.6%) and human (86.5%). The alpha-chain Hb gene encoded a 142-amino acid protein that could be identified with the homology of alpha-chain Hb protein in sheep (98%), goat (96%), cattle (91%) and human (87%). However, 18 alternations were detected when compared with the alpha-chain Hb gene in human, and 2 in sheep. Moreover, the alterations of a117 GluAsp and $\alpha$132 AsnSer in important regions were noted in human and sheep, respectively. Phylogenetic analysis suggested that the structure of alpha-chain Hb was highly similar to that in sheep. This study provided essential information for elucidating the possible roles of hemoglobin in adapting to extremely high altitude in Pantholops hodgsonii.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.19-25
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• 1985

한국(韓國), 대만(臺灣), 일본(日本)에서 사육(飼育)하고 있는 꽃사슴(Cervus nippon) 129두(頭)와 물사슴(Cervus unicolor) 7두(頭)에서 hemoglobin, transferrin, albumin carbonic anhydrase, slow-${\alpha}_2$ 및 amylase형(型)을 전기영동(電氣泳動)에 의(依)하여 분석(分析)한 결과(結果) 다음과 같은 결론(結論)을 얻었다. 1. cellulose acetate에 의(依)한 전기영동(電氣泳動)이 starch gel에 의(依)한 전기영동(電氣泳動)보다 hemoglobin형(型) 분리(分離)에 있어서 더 간편하며 시간이 적게 걸리고, 선명(鮮明)할 뿐만 아니라 영구보존(永久保存)이 가능(可能)하다. 2. 꽃사슴의 hemoglobin형(型)은 $Hb^F$, $Hb^{FS}$, $Hb^S$형(型)으로 분리(分離)되었으나 물사슴에 있어서는 전부(全部) $Hb^F$형(型)으로 나타났다. 3. hemoglobin ${\beta}$ chain은 4가지형(型) 즉 ${\beta}$-1, ${\beta}$-2, ${\beta}$-3 및 ${\beta}$-4로 분리(分離)되었다. 4. hemoglobin ${\alpha}$ chain은 ${\alpha}_1$과 ${\alpha}_1{\alpha}_2$형(型)으로 분리(分離)되었다. 5. slow-${\alpha}_2$형(型)은 A형(型)과 AB형(型)으로 분리(分離)되었으며, 꽃사슴에 있어서는 AB형(型)이 12% 출현(出現)하였으나 물사슴에서는 전부(全部) A형(型)으로 AB형(型)은 없었다. 6. albumin형(型)에서는 F형(型)과 S형(型)으로 분리(分離)되었으며 꽃사슴은 전부(全部) F형(型)이였고, 물사슴은 전부(全部) S형(型)이였다. 7. transferrin형(型), carbonic anhydrase형(型) 및 amylase형(型)은 전부(全部) 각각(各各) 1종류(種類)의 형(型)이였다.

• BMB Reports
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• v.38 no.1
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• pp.115-119
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• 2005

Replacement of valine by tryptophan or tyrosine at position $\alpha$96 of the $\alpha$ chain ($\alpha$96Val), located in the ${\alpha}_1{\beta}_2$ subunit interface of hemoglobin leads to low oxygen affinity hemoglobin, and has been suggested to be due to the extra stability introduced by an aromatic amino acid at the $\alpha$96 position. The characteristic of aromatic amino acid substitution at the $\alpha$96 of hemoglobin has been further investigated by producing double mutant r Hb ($\alpha$42Tyr$\rightarrow$ Phe, $\alpha$96Val$\rightarrow$Trp). r Hb ($\alpha$42Tyr$\rightarrow$Phe) is known to exhibit almost no cooperativity in binding oxygen, and possesses high oxygen affinity due to the disruption of the hydrogen bond between $\alpha$42Tyr and $\beta$99Asp in the ${\alpha}_1{\beta}_2$ subunit interface of deoxy Hb A. The second mutation, $\alpha$96Val$\rightarrow$Trp, may compensate the functional defects of r Hb ($\alpha$42Tyr$\rightarrow$Phe), if the stability due to the introduction of trypophan at the $\alpha$96 position is strong enough to overcome the defect of r Hb ($\alpha$42Tyr$\rightarrow$Phe). Double mutant r Hb ($\alpha$42Tyr$\rightarrow$Phe, $\alpha$96Val$\rightarrow$Trp) exhibited almost no cooperativity in binding oxygen and possessed high oxygen affinity, similarly to that of r Hb ($\alpha$42Tyr$\rightarrow$Phe). $^1$H NMR spectroscopic data of r Hb ($\alpha$42Tyr$\rightarrow$Phe, $\alpha$96Val$\rightarrow$Trp) also showed a very unstable deoxy-quaternary structure. The present investigation has demonstrated that the presence of the crucible hydrogen bond between $\alpha$42Tyr and $\beta$99Asp is essential for the novel oxygen binding properties of deoxy Hb ($\alpha$96Val$\rightarrow$Trp).

• Parasites, Hosts and Diseases
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• v.53 no.3
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• pp.265-270
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• 2015

Hemoglobinopathy and malaria are commonly found worldwide particularly in malaria endemic areas. Thalassemia, the alteration of globin chain synthesis, has been reported to confer resistance against malaria. The prevalence of thalassemia was investigated in 101 malaria patients with Plasmodium falciparum and Plasmodium vivax along the Thai-Myanmar border to examine protective effect of thalassemia against severe malaria. Hemoglobin typing was performed using low pressure liquid chromatography (LPLC) and ${\alpha}$-thalassemia was confirmed by multiplex PCR. Five types of thalassemia were observed in malaria patients. The 2 major types of thalassemia were Hb E (18.8%) and ${\alpha}$-thalassemia-2 (11.9%). There was no association between thalassemia hemoglobinopathy and malaria parasitemia, an indicator of malaria disease severity. Thalassemia had no significant association with P. vivax infection, but the parasitemia in patients with coexistence of P. vivax and thalassemia was about 2-3 times lower than those with coexistence of P. falciparum and thalassemia and malaria without thalassemia. Furthermore, the parasitemia of P. vivax in patients with coexistence of Hb E showed lower value than coexistence with other types of thalassemia and malaria without coexistence. Parasitemia, hemoglobin, and hematocrit values in patients with coexistence of thalassemia other than Hb E were significantly lower than those without coexistence of thalassemia. Furthermore, parasitemia with coexistence of Hb E were 2 times lower than those with coexistence of thalassemia other than Hb E. In conclusion, the results may, at least in part, support the protective effect of thalassemia on the development of hyperparasitemia and severe anemia in malaria patients.

• Journal of Microbiology and Biotechnology
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• v.3 no.2
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• pp.123-128
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• 1993

The polyesters (polyhydroxyalkanoates; PHAs) production capability in a two-step cultivation of Alcaligenes eutrophus H16(ATCC 17699) was investigated by using various organic carbon sources. The carbon sources used included linear $C_2~C_10$ monocarboxylic acids, $C_3~C_10$ dicarboxylic acids, crotonic acid, and several linear vicinal and $\omega$-diols. The polyesters synthesized were characterized by 500 MHz $^1 H-NMR$ spectroscopy, intrinsic viscosity$[\eta]$ measurement in chloroform and differential scanning calorimetry (DSC). The PHAs synthesis data showed that the use of C-odd ($C_3, C_5, and C_7$) monocarboxylic acids resulted in poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(P(3HB-co-3HV) (3HV content ranging 40 to 70 mol%) while the use of $C_9$ substrate gave the copolyester containing only 4 mol% of 3HV. All culture products obtained on $C_3$~C$_{10}$ dicarboxylic acids gave exclusively P(3HB). 500 MHz $^1 H-NMR$ analysis showed that all polyesters synthesized generally contained 1~2 mol% 3HV even for the unrelated substrates such as the carboxylic acids with even number of carbon. When $\alpha, \omega$-diols with even number of carbon were used as substrates, 4-hydroxybutyrate(4HB) was inserted into the polyester chain composed of P(3HB-co-4HB). Vicinal diols were generally not utilized by the bacterium for polyester production.n.

• Parasites, Hosts and Diseases
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• v.56 no.2
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• pp.167-173
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• 2018

Malaria is one of the most important public health problems in tropical areas on the globe. Several factors are associated with susceptibility to malaria and disease severity, including innate immunity such as blood group, hemoglobinopathy, and heme oxygenase-1 (HO-1) polymorphisms. This study was carried out to investigate association among ABO blood group, thalassemia types and HO-1 polymorphisms in malaria. The malarial blood samples were collected from patients along the Thai-Myanmar border. Determination of ABO blood group, thalassemia variants, and HO-1 polymorphisms were performed using agglutination test, low pressure liquid chromatography and polymerase chain reaction, respectively. Plasmodium vivax was the major infected malaria species in the study samples. Distribution of ABO blood type in the malaria-infected samples was similar to that in healthy subjects, of which blood type O being most prevalent. Association between blood group A and decreased risk of severe malaria was significant. Six thalassemia types (30%) were detected, i.e., hemoglobin E (HbE), ${\beta}$-thalassemia, ${\alpha}$-thalassemia 1, ${\alpha}$-thalassemia 2, HbE with ${\alpha}$-thalassemia 2, and ${\beta}$-thalassemia with ${\alpha}$-thalassemia 2. Malaria infected samples without thalassemia showed significantly higher risk to severe malaria. The prevalence of HO-1 polymorphisms, S/S, S/L and L/L were 25, 62, and 13%, respectively. Further study with larger sample size is required to confirm the impact of these 3 host genetic factors in malaria patients.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.7
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• pp.929-937
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• 2016

The purpose of this study was to investigate the effect of Acanthopanax senticosus extract (ASE) (ethanol : DW=1:1, v/v) on inhibition of type 2 diabetes using an OLETF rat model via regulation of HbA1c and AGEs levels. Supplementation with ASE 0.1% and 0.5% effectively lowered levels of glucose, insulin, oral glucose tolerance test, and Homa-insulin resistance, suggesting reduced insulin resistance. Blood levels of HbA1c and AGEs were significantly reduced in a dose-dependent manner. As oxidative stress plays a key role in accelerating production of HbA1c and AGEs, which worsen symptoms of type 2 diabetes, levels of malonaldehyde and pro-inflammatory cytokines were measured. Lipid peroxidation in both blood and liver tissues was significantly reduced, and induction of pro-inflammatory cytokines interleukin-${\beta}$ and tumor necrosis factor-${\alpha}$, which elevate production of HbA1c and AGEs, was inhibited (P<0.05). To evaluate the possible cellular events after AGEs receptor activation, genetic expression of protein kinase C (PKC)-${\delta}$ and transforming growth factor (TGF)-${\beta}$ was measured by real-time polymerase chain reaction. Supplementation with both ASE 0.1% and 0.5% significantly inhibited mRNA expression of PKC-${\delta}$ and TGF-${\beta}$, indicating that ASE may have beneficial effects on preventing insulin-resistant cells or tissues from progressing to diabetic complications. Taken together, ASE has potential to improve type 2 diabetes by inhibiting insulin resistance and protein glycosylation, including production of HbA1c and AGEs. Anti-oxidative activities of ASE are a main requisite for reducing production of HbA1c and AGEs and are also related to regulation of the PKC signaling pathway, resulting in suppression of TGF-${\beta}$, which increases synthesis of collagen, prostaglandin, and disease-related proteins.